A simple and highly effective method for the stable transduction of uncultured porcine hepatocytes using lentiviral vector

Gene therapy is an attractive approach for the treatment of a wide spectrum of liver diseases. Lentiviral vectors allow the stable integration of transgenes into the genome of nondividing differentiated cells including hepatocytes and could provide long-lasting expression of a therapeutic gene. To develop such approaches, preclinical studies in large animal models such as pigs are necessary to evaluate the feasibility and safety of stable lentiviral integration and long-term vector expression. In addition, effective lentivector-mediated gene transfer onto porcine hepatocytes could advance in cell-based therapies for acute liver failure. To investigate this issue, porcine hepatocytes were transduced in suspension immediately after their isolation in University of Wisconsin (UW) solution containing vitamin E. Up to 80% of hepatocytes stably expressed a GFP transgene after a single exposure to lentiviral vector coding for GFP under the control of either liver-specific or ubiquitous promoters. Moreover, porcine hepatocytes cryopreserved in UW solution containing fetal bovine serum, dimethyl sulfoxide, and vitamin E remained highly transducible with lentiviral vector after thawing. When thawed, transduced in suspension, and immediately transplanted into the spleen of immunodeficient mice, ex vivo lentivirally transgene marked xenogeneic hepatocytes were detected in murine liver. We demonstrated that porcine hepatocytes are highly susceptible to lentiviral vector and describe an easy methodology to efficiently, rapidly, and stably introduce transgenes into uncultured porcine hepatocytes.     

Recombinant activated factor VII decreases bleeding without increasing arterial thrombosis in rabbits

PURPOSE: To compare the effects of recombinant activated factor VII (rFVIIa) and platelet-rich plasma (PRP) in an experimental model of bleeding and arterial thrombosis. METHODS: The Folts model was used in 60 rabbits. After anesthesia, the carotid artery was exposed and a 75% stenosis was induced. A compression injury of the artery triggered a series of cyclic flow reductions (CFRs). After counting baseline CFRs, animals were assigned randomly to one of four groups (n = 15 in each): control, PRP, rFVIIa and placebo. Control animals received 10 mL.kg(-1) of saline while 10 mL.kg(-1) of a hydroxyethyl starch solution (200,000/6%/0.5) were infused in the three other groups. CFRs were measured again, followed by treatment with PRP, rFVIIa or placebo and by a final measurement of CFRs. At the end of each observation period, an ear immersion bleeding time (BT) was measured and a blood sample was drawn for the evaluation of hematological variables. Microvascular bleeding was evaluated at the end of the experiment in grams of blood shed from liver and spleen sections. Results are presented as median (range). RESULTS: rFVIIa shortened the BT and decreased microvascular bleeding as compared with placebo [60 (35-100) sec vs 110 (50-140) sec, P = 0.0019 and 9 (4-24) g vs 17 (5-28) g, P = 0.002, respectively]. rFVIIa did not increase CFRs [3(0-9) vs |(0-5), P = 0.11]. CONCLUSION: rFVIIa led to a decrease in BT and microvascular bleeding but did not significantly affect arterial thrombosis in rabbits.     

Meaning in life and mastery mediate the relationship of negative reminiscence with psychological distress among older adults with mild to moderate depressive symptoms

To understand the adaptive value of reminiscence, a mediational model of reminiscence was tested in a sample of older adults with mild to moderate depressive symptoms. Using structural equation modeling, we investigated if psychological resources (mastery and meaning in life) mediate the relation between reminiscence (positive: identity construction and problem solving; and negative: bitterness revival and boredom reduction) and psychological distress (depressive symptoms and anxiety symptoms). A total of 202 older Dutch adults living in the community participated in this study. The present study consisted of baseline measurements of a randomized controlled trial that evaluated the effectiveness of a life-review therapy intervention on depression. Results showed that psychological resources fully mediated the relation between negative reminiscence and psychological distress. Specifically, negative reminiscing is related to decreased psychological distress through meaning in life and sense of mastery. The study contributes to current knowledge on the relation between reminiscence and mental health, both empirically and clinically. It helps to increase understanding of how reminiscence is related to psychological distress, especially in depressed older adults, and the relative importance of psychological resources, i.e., mastery and meaning in life. From a clinical perspective, these findings suggest the usefulness of focusing on strengthening psychological resources in therapeutic reminiscence-based strategies for older adults with depressive symptoms.     

First inappropriate implantable cardioverter defibrillator therapy is often due to inaccurate device programming: analysis of the French OPERA registry

AIMS: Inappropriate therapy delivered by implantable cardioverter defibrillators (ICDs) remains a challenge. The OPERA registry measured the times to, and studied the determinants of, first appropriate (FAT) and inappropriate (FIT) therapies delivered by single-, dual- and triple-chamber [cardiac resynchronization therapy defibrillator (CRT-D)] ICD. METHODS AND RESULTS: We entered 636 patients (mean age = 62.0 ± 13.5 years; 88% men) in the registry, of whom 251 received single-, 238 dual-, and 147 triple-chamber ICD, for primary (30.5%) or secondary (69.5%) indications. We measured times to FAT and FIT as a function of multiple clinical characteristics, examined the effects of various algorithm components on the likelihood of FAT and FIT delivery, and searched for predictors of FAT and FIT. Over 22.8 ± 8.8 months of observation, 184 patients (28.9%) received FAT and 70 (11.0%) received FIT. Ventricular tachycardia (VT) was the trigger of 88% of FAT, and supraventricular tachycardia was the trigger of 91% of FIT. The median times to FIT (90 days; range 49-258) and FAT (171 days; 50-363) were similar. The rate of FAT was higher (P <0.001) in patients treated for secondary than primary indications, while that of FIT were similar in both groups. Out of 57 analysable FIT, 27 (47.4%) could have been prevented by fine tuning the device programming like the sustained rate duration or the VT discrimination algorithm. CONCLUSIONS: First inappropriate therapy occurred in 11% of 636 ICD recipients followed for ～2 years. Nearly 50% of FIT could have been prevented by improving device programming.     

Balance training in ataxic neuropathies. Effects on balance and gait parameters

A rehabilitation program including foot sensory stimulation, balance and gait training with limited vision was followed by 30 patients with ataxic neuropathy in order to stimulate multi-sensory compensation in a no-controlled and no-blinded study. Ataxic neuropathy was graded by a pallesthetic score. The evaluation of patients and healthy subjects was performed with clinical tests (Berg Balance Scale, Functional Reach Test and Timed up and Go test) and instrumental tests for balance (force platform) and gait (Locometre). All patients exhibited impairments in balance and gait parameters compared to control group values. A high pallesthetic score correlated with increased sway area when standing with the eyes open on a firm surface. At the end of the training program, significant changes were observed in balance control assessed using the three clinical tests (Wilcoxon test, p < 0.001). A tendency towards a reduction of the Romberg sign was noticed and limited changes were observed after training in instrumental tests for balance and for gait parameters. Age induced some limitations in balance and gait parameters but had no effect on training results. These results show that ataxic patients are impaired in balance and gait but can improve clinical balance parameters following training with a multisensory approach without limitation due to age or degree of sensory impairment. Only limited correlations were noticed between the pallesthetic score and some balance parameters, suggesting that various levels of compensation occur in these patients. The effectiveness of this training program has to be evaluated in the future in a controlled study to ascertain the contribution of the placebo effect in these data.