Clinical and pharmacological significance of alpha2-adrenoceptor polymorphisms in cardiovascular diseases

The alpha2-adrenoceptors (alpha2-ARs) are receptors for endogenous catecholamines (norepinephrine and epinephrine) that mediate a number of physiological and pharmacological responses such as hypotension and sedation. Three distinct subtypes, denoted alpha2A-, alpha2B- and alpha2C-AR, have been characterized and cloned. Employment of mutation screening in the study of human populations from various ethnic backgrounds has shown that alpha2-AR genes are polymorphic. The functional and biochemical consequences of these polymorphisms have been analyzed by expressing the wild-type receptors and their respective genetic variants in heterologous systems such as CHO and COS-7 cells. Changes include alteration in G-protein coupling and in agonist-promoted receptor phosphorylation and desensitization. Case-control and population-based studies have shown clinical association with cardiovascular risk. Further investigation of the genetic variants in specialized cells and transgenic animals will provide the molecular basis of cardiovascular disease and may reveal alpha2-AR variants as potential targets for selective pharmacological interventions.     

Effect of an experimental stenosis in the porcine descending thoracic aorta

OBJECTIVE: To investigate the alterations of mechanical properties in pre- (A) and post- (B) stenotic aortic regions after an experimentally induced stenosis in the descending thoracic aorta. METHODS: Eight healthy, normalipemic and normotensive Landrace pigs were subjected to thoracotomy under sterile conditions. In the upper segment of the descending thoracic aorta a circumferential symmetric constriction 5 mm in length was imposed and stabilized; thus, a 15-20 mm Hg pressure gradient was established. The pressure gradient was verified via catheterization of the vessel with pressure tip catheters. Pre- and poststenotic hemodynamic disturbances were recorded by the use of a bidirectional Doppler flowmeter. Pressure and flow measurements were carried out before, 10 min after, and 90 days after the creation of the stenosis. Euthanasia was performed after 90 days, and the descending thoracic aorta removed. In the A and B regions serial sections of aorta (5 mm in length) create appropriate aortic "rings," to test in a uniaxial tension device, in order to determine the mechanical properties of the vessel. Histological analysis was performed, so as to estimate the content (%) of collagen and elastin fibers within the aortic wall. Eight sham-operated pigs were used as controls. RESULTS: Reverse blood flow was recorded at both the A and B sites, and was maintained until euthanasia. Reverse flow in the poststenotic region B was greater than that in the prestenotic region. Mechanical analysis showed that the aortic wall in A and B regions became stiffer particularly at high strains (P < 0.05). Histological analysis indicated that the percentage of elastin fibers remained almost the same in both regions while the percentage of collagen fibers increased considerably, especially in the B region (P < 0.05). CONCLUSIONS: A nonhemodynamically significant experimental stenosis located at the level of the descending thoracic aorta induced reverse blood flow before and after the stenosis. The higher the reverse flow, the more collagen fibers were produced and the stiffer the aortic wall. Since hypercholesterolemia can be ruled out as a hardening factor in the present study, disturbed flow seems to be an independent factor which activates fibroblasts to overproduce collagen and to eventually reduce the aorta's compliance.     

Modulation of differentiation and mineralization of marrow stromal cells cultured on biomimetic hydrogels modified with Arg-Gly-Asp containing peptides

We synthesized biomimetic hydrogels modified with an osteopontin-derived peptide (ODP) and used them as a substrate for in vitro culture of marrow stromal cells (MSCs) to investigate the effect of the biomimetic surface on differentiation of MSCs into osteoblasts. Proliferation and biological assays for 16 days proved that MSCs became differentiated into osteoblasts secreting osteogenic phenotypic markers such as alkaline phosphatase (ALP), osteopontin, and mineralized calcium. In addition, there was an additive effect of the cell-binding peptide on differentiation and mineralization of MSCs cultured in the presence of soluble osteogenic supplements in cell culture media. For example, calcium content at day 16 on peptide-modified hydrogels was significantly higher than on tissue culture polystyrene. Two general trends were observed: (1) proliferation of MSCs decreased as the amount of differentiation markers increased, and (2) higher peptide concentrations accelerated the differentiation of MSCs. On the hydrogel modified with ODP, ALP activity exhibited a maximum value of 36.7 +/- 4.2 pmol/cell/h at day 10 for the concentration of 2 micromol/g while the culture time needed for maximum ALP activity occurred on day 13 for the lower concentrations. On the same hydrogel, the calcium content at day 10 was 21.4 +/- 2.3 ng/cell for the peptide concentration of 2 micromol/g and 1.0 +/- 0.3 ng/cell for 1.0 micromol/g. We used Gly-Arg-Gly-Asp-Ser (GRGDS) for modification of the hydrogel as a comparison to the results with ODP. However, osteoblast development was not significantly affected by the nature of the binding peptide sequences. These results suggest that MSC function can be modulated by variation of the peptide concentration in biomimetic hydrogels used for scaffold-based bone tissue engineering.     

A randomized, controlled, clinical trial of activity therapy for apathy in patients with dementia residing in long-term care

BACKGROUND: Apathy is a common symptom in patients with dementia and has adverse consequences for patients and caregivers. Most treatments for apathy, particularly non-pharmacologic interventions, have not been evaluated in controlled trials. OBJECTIVES: This study evaluated the efficacy of a kit-based activity intervention, compared to a time and attention control (one-on-one meetings with an activity therapist) in reducing apathy and improving quality of life in 37 patients with dementia. METHODS: The design was a randomized, controlled, partially masked clinical trial. All outcome measures were administered at baseline and follow-up. The primary outcome measure was the apathy score of the Neuropsychiatric Inventory (NPI). Other outcome measures were the NPI total score, the Alzheimer Disease Related Quality of Life scale(ADQRL), and the Copper Ridge Activity Index (CRAI). RESULTS: There was a significant reduction in NPI apathy scores in both treatment groups. The only significant difference between the two treatment groups was a modest advantage for the control intervention on the CRAI cueing subscale (p = 0.027), but not on the other CRAI subscales. There was also a greater within group improvement in quality of life ratings in the control intervention (p=0.03). CONCLUSIONS: Despite the substantial improvement in apathy scores during the course of the study, there was no clear advantage to the reminiscence-based intervention over the time and attention, one-on-one control intervention. More research is needed to develop specific behavioral interventions for apathy in patients with dementia.     

Outbreak of infections caused by Enterobacter cloacae producing the integron-associated beta-lactamase IBC-1 in a neonatal intensive care unit of a Greek hospital

Nineteen of 27 ceftazidime-resistant Enterobacter cloacae isolates from a neonatal intensive care unit in Thessaloniki, Greece, had genes coding for the novel extended-spectrum beta-lactamase IBC-1; 18 of those 19 harbored similar conjugative plasmids and belonged to two distinct genetic lineages. A synergy test with ceftazidime and imipenem enabled us to identify five unrelated bla(IBC-1)-carrying E. cloacae isolates from other wards of the hospital. It seems that this integron-associated gene is capable of dispersing both by clonal spread and by gene dissemination.     

Plasma pituitary adenylate cyclase activating polypeptide (PACAP) levels in chronic hepatitis B patients under lamivudine treatment

OBJECTIVE: Lamivudine is a nucleoside analogue with potent antiviral activity against hepatitis B virus (HBV). Plasma pituitary adenylate cyclase activating polypeptide (PACAP) is a multifunctional neuropeptide that is produced within the lymphoid microenvironment and induces the production of Th2-type cytokines. The aim of our study was to investigate the possible alterations of plasma PACAP-38 levels in chronic hepatitis B (CHB) patients during lamivudine treatment and to compare them with biochemical, virological and histological data. METHODS: Plasma PACAP-38 levels were measured using competitive radio-immune analysis (RIA) in 25 CHB patients before and after completion of a 52-week lamivudine treatment period and in 22 healthy blood donors. Biochemical evaluation was done at baseline and every three months during treatment. Virological evaluation (HBV-DNA) was performed at baseline and at weeks 24 and 52 of treatment. Baseline liver histology was assessed for all patients at the beginning and at week 52 of the study for histological comparison with the pretreatment biopsy, according to the Ishak scoring system. Statistical evaluation of data was done using analysis of variance and Student's t-test. RESULTS: Virological breakthrough was observed in seven (28%) patients at week 52 of treatment. Histological improvement was observed in 21 (84%) CHB patients, despite the emergence of tyrosine-methionine-aspartate-aspartate (YMDD) mutations. Plasma PACAP-38 levels were significantly lower in CHB patients at baseline than in healthy blood donors. Significant elevation of plasma peptide levels was observed in CHB patients after the completion of lamivudine treatment period, even in the subgroup of those who exhibited YMDD variants. CONCLUSION: The elevation of plasma PACAP-38 levels in treated CHB patients following lamivudine-induced elimination of viraemia suggests a possible alteration of T-cellular immune response, resulting in biochemical and histological remission of liver disease, even in patients who exhibited virological breakthrough.     

Cell adhesion on poly(propylene fumarate-co-ethylene glycol) hydrogels

We synthesized poly(propylene fumarate-co-ethylene glycol) block copolymers [P(PF-co-EG)] that were crosslinked to form hydrogels and investigated the effect of copolymer composition on cell adhesion to the hydrogels. These copolymers were water soluble when the molar ratio of ethylene glycol repeating unit to propylene fumarate repeating unit was higher than 4.4. The water content of swollen hydrogels increased from 29 to 63% and the water contact angle decreased from 38 to 21 degrees as the molar ratio increased from 0.6 to 4.4. No significant change in either property was observed for ratios higher than 4.4. In a cell adhesion assay under serum-free conditions, the number of adherent platelets and smooth muscle cells decreased from 21 to 2% and from 78 to 20% of the initial seeding density, respectively, as the molar ratio increased from 0.6 to 7.8. Adherent smooth muscle cells did not spread on the hydrogels of the compositions tested. Adherent platelets did not show any filopodia. These results suggest that the hydrophilicity of P(PF-co-EG) hydrogels is one of the factors affecting cell adhesion, and that copolymer modification may be required for enhancing cell adhesion for an application involving the copolymers as in situ crosslinkable cell carriers.