Synergistic effects of deleting multiple nonessential elements in nonreplicative HSV-1 BAC genomic vectors play a critical role in their viability.

Nonreplicative Herpes simplex virus type-1 (HSV-1) genomic vectors have already entered into clinical trials for neurological gene therapy thanks to their scalable growth in permissive cells. However, the small transgene capacity of this type of HSV-1 vectors currently used in the clinic represents an important limiting factor as a gene delivery system. To develop high-capacity nonreplicative genomic HSV-1 vectors, in this study we have characterized a series of multiply deleted mutants which we have constructed in bacterial artificial chromosomes (BACs), removing up to 24 kb of unstable or dispensable genomic sequences to allow insertion of transgenes up to this size. We show that synergistic effects of deletions of: the HSV-1 replication origins oriS and oriL, the HSV-1 internal repeat region, the remaining ICP4 gene copy and the genes encoding for ICP27, UL56, UL55, can severely reduce the growth of these HSV-1 vectors. Given that several of these elements have been characterized as 'non-essential' for viral growth in cell culture by single-deletion experiments of wild-type HSV-1, our study highlights the need to re-evaluate their functional contribution in the context of multiply deleted nonreplicative HSV-1 genomic vectors. Our BAC mutants described here can serve as useful starting platforms to accelerate HSV-1 vector development.

Gene therapy approach to FAP: in vivo influence of T119M in TTR deposition in a transgenic V30M mouse model.

Familial amyloidotic polyneuropathy (FAP) is a neurodegenerative disorder characterized by extracellular deposition of amyloid fibrils composed by mutated transthyretin (TTR) mainly in the peripheral nervous system. At present, liver transplantation is still the standard treatment to halt the progression of clinical symptoms in FAP, but new therapeutic strategies are emerging, including the use of TTR stabilizers. Here we propose to establish a new gene therapy approach using adeno-associated virus (AAV) vectors to deliver the trans-suppressor TTR T119M variant to the liver of transgenic TTR V30M mice at different ages. This TTR variant is known for its ability to stabilize the tetrameric protein. Analysis of the gastrointestinal tract of AAV-treated animals revealed a significant reduction in deposition of TTR non-fibrillar aggregates in as much as 34% in stomach and 30% in colon, as well as decreased levels of biomarkers associated with TTR deposition, namely the endoplasmic reticulum stress marker BiP and the extracellular matrix protein MMP-9. Moreover, we showed with different studies that our approach leads to an increase in tetrameric and more stable forms of TTR, in favor of destabilized monomers. Altogether our data suggest the possibility to use this gene therapy approach in a prophylactic manner to prevent FAP pathology.

A randomized double-blind clinical trial with two different doses of arginine enhanced enteral nutrition in postsurgical cancer patients.

OBJECTIVES: Patients with head and neck cancer undergoing surgery have a high incidence of postoperative complications. The aim of our study was to investigate whether postoperative nutrition of head and neck cancer patients, using an enteral high dose of arginine (20 g per day) vs a medium dose (12.3 g per day) could improve nutritional variables as well as clinical outcome. MATERIAL AND METHODS: A population of 115 patients with oral and laryngeal cancer was enrolled. At surgery patients were randomly allocated to two groups: group I (58 patients) received an enteral diet supplements with a high dose of arginine (20 g per day) and group II (57 patients) received an isocaloric, isonitrogenous enteral formula with a medium dose of arginine (12.3 g per day). RESULTS: Gastrointestinal tolerance (diarrhea) of both formulas was good (3.44% group I and 3.51% group II: ns). The postoperative infections complications were similar in both groups (8.6% group I and 12.2% group II: ns). Fistula was less frequent in enriched nutrition group (3.4% group I and 10.5% group II: p = 0.006). The length of postoperative stay was similar in both groups (27.2 +/- 17.8 days in group I vs 25.7 +/- 18.8 days in group II: ns). CONCLUSIONS: Enriched arginine formula improves fistula wound complications in postoperative head and neck cancer patients. Our results suggest that these patients could benefit from a high dose of arginine enhanced enteral formula.

G 1359A polymorphism of the cannabinoid receptor gene (CNR1) and clinical results of biliopancreatic diversion.

BACKGROUND: Bariatric surgery is the most effective long-term treatment for morbid obesity, reducing obesity-associated co-morbidities. We decide to investigate the role of the polymorphism (G1359A) of the cannabinoid (CB)1 receptor gene on clinical outcomes 1 year after biliopancreatic diversion in morbidly obese patients. DESIGN: A sample of 66 morbidly obese patients (BMI >40 kg/m2) were operated. Weight, fat mass, blood pressure, basal glucose, triacylglycerols, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol were measured at basal visit and at each visit. The frequency of metabolic comorbidities was recorded at each visit. RESULTS: Thirty-nine patients (59.1%) had genotype G1359G (wild type group) and 27 (40.9%) patients had genotype G1359A (mutant type group). In wild and mutant type groups, body mass index, weight, waist circumference, systolic blood pressure, glucose, total cholesterol, low-density lipoprotein cholesterol and triacylglycerols concentrations decreased, without statistical between genotype groups. Initial weight percent loss at 1 year of follow up was similar in both genotypes (33.1% vs 33.6%; ns). CONCLUSION: The present study demonstrates that polymorphism G1359A in the CB1 receptor did not have a significant effect on biochemical and anthropometric improvements after biliopancreatic diversion surgery.

Effect of three different doses of arginine enhanced enteral nutrition on nutritional status and outcomes in well nourished postsurgical cancer patients: a randomized single blinded prospective trial.

OBJECTIVE: Patients with head and neck cancer undergoing surgery have a high occurrence of postoperative complications. The aim of our study was to investigate whether postoperative nutrition of head and neck cancer patients, using an enhanced enteral formula with three different doses of arginine could improve nutritional variables as well as clinical outcome, depending of arginine dose. PATIENTS AND METHODS: A population of 84 patients with oral and laryngeal cancer was enrolled. At surgery patients were randomly assigned to three different treatment groups, each one containing at less 28 patients. Group I (28 patients) received an enteral diet supplements with a low physiological dose of arginine (5.7 g per day), group II (28 patients) received an isocaloric, isonitrogenous enteral formula with a medium dose of arginine (12.3 g per day) and group III (28 patients) received an isocaloric, isonitrogenous enteral formula with a high dose of arginine (18.9 g per day). RESULTS: The length of postoperative stay had a trend to be better with high dose of arginine received (31.9 ± 17.2 days in group I vs 27.8 ± 15.2 days in group II vs 24.8 ± 18.3 days in group III: p = 0.034). No differences were detected in postoperative infections complications and diarrhea. Fistula was less frequent in groups II and III than I (10.7% group I vs 3.6% group II vs 3.6% group III: p = 0.033), The length of postoperative stay had a trend to be better with high dose of arginine received (31.9 ± 17.2 days in group I vs 27.8 ± 15.2 days in group II vs 24.8 ± 18.3 days in group III: p = 0.034). CONCLUSIONS: Our results suggest that these patients could benefit from a high dose of arginine enhanced enteral formula to decrease length of hospital stay and fistula wound complications.

Clinical effects of a w3 enhanced powdered nutritional formula in postsurgical ambulatory head and neck cancer patients.

BACKGROUND: Patients with head and neck cancer undergoing surgery have a high risk of nutritional complications. OBJECTIVE: The aim of our study was to investigate the influence of an oral w3 enriched immunoenhanced powdered formula in nutritional and biochemical parameters in postsurgical ambulatory patients with head and neck tumor. DESIGN: A population of 33 ambulatory postsurgical patients with oral and laryngeal cancer was enrolled. At Hospital discharge postsurgical head and neck cancer patients were asked to consume two units per day of a w3 enriched immunoenhanced powdered formula for a twelve week period. RESULTS: The mean age was 61.3 ± 9.1 years (6 female/27 males). Duration of supplementation was 95.9 ± 21.1 days. A significant increase of albumin and transferrin levels was observed, in total group and in patients undergoing radiotherapy and without it. No differences were detected in weight and other anthropometric parameters in total group and in patients with radiotherapy during the protocol. Nevertheless, patients without radiotherapy showed a significant improvement of BMI; weight, fat free mass and fat mass. CONCLUSIONS: At dose used, an omega 3 enriched powdered formula improved seric protein levels in ambulatory postoperative head and neck cancer patients. Improvement of weight, fat mass and fat free mass was observed in patients whom not received radiotherapy during the follow up.

Antecedentes: Los pacientes con tumores de cabeza y cuello sometidos a cirugía presentan un alto riesgo de complicaciones nutricionales. Objetivo: El principal objetivo de nuestro trabajo fue evaluar la influencia de un suplemento oral en polvo inmunoenriquecido con ácidos grasos omega 3 en pacientes postquirúrgicos ambulatorios con tumores de cabeza y cuello. Diseño: Una muestra de 33 pacientes postquirúrgicos ambulatorios con tumores de cabeza y cuello fue evaluada. Tras el alta hospitalaria, los pacientes recibieron dos envases al día de un suplemento inmunoenriquecido con omega 3 en polvo durante 12 semanas. Resultados: La edad media fue de 61,3 ± 9,1 años (6 mujeres/27 varones). La duración media de la suplementación fue de 95,9 ± 21,1 días. Se detectó un aumento significativo en los niveles de albúmina y transferrina en los pacientes del grupo global, en los que recibieron radioterapia y en los que no al recibieron. El peso y los parámetros antropométricos no mejoraron en el grupo global ni en los que recibieron radioterapia. Sin embargo los pacientes que no recibieron radioterapia tuvieron un aumento de la masa magra, peso y masa grasa Conclusiones: A la dosis usada, la formula en polvo enriquecida en omega 3 mejoró lo niveles de proteínas séricas. Por otra parte los pacientes que no recibieron radioterapia durante el seguimiento presentaron un aumento del peso, masa grasa y masa magra.

Effects of C358A missense polymorphism of the endocannabinoid degrading enzyme fatty acid amide hydrolase on weight loss and cardiovascular risk factors 1 year after biliopancreatic diversion surgery.

BACKGROUND: Bariatric surgery is the most effective long-term treatment of morbid obesity and also results in a reduction of obesity-associated co-morbidities. We investigated the role of the polymorphism (C358A) of the fatty acid amide hydrolase gene on the clinical outcomes 1 year after biliopancreatic diversion in morbidly obese patients. METHODS: A total of 67 morbidly obese patients (body mass index >40 kg/m(2)) underwent biliopancreatic diversion. Their weight, blood pressure, basal glucose, triglycerides, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol were measured at the baseline visit and at each follow-up visit. The frequency of the metabolic co-morbidities was recorded at each visit. RESULTS: Of the 67 patients, 46 (68.7%) had genotype C358C (wild-type group) and 21 (10.3%) had genotype C358A (mutant-type group). In the wild- and mutant-type groups, the body mass index, weight, waist circumference, systolic blood pressure, and glucose, total cholesterol, low-density lipoprotein cholesterol, and triglyceride concentrations decreased, without statistical significance between the 2 groups. The initial percentage of weight loss at 9 months and 1 year of follow-up was greater in the mutant-type group (9 months, 22.1% versus 28.8%, P <.05; and 1 year, 28.3% versus 36.4%, P <.05). CONCLUSION: The allele A358 of fatty acid amide hydrolase was associated with a better initial percentage of excess weight loss 9 and 12 months after biliopancreatic diversion.

Influence of an immuno-enhanced formula in postsurgical ambulatory patients with head and neck cancer.

BACKGROUND: Patients with head and neck cancer undergoing surgery have a high incidence of ambulatory postoperative complications. OBJECTIVE: The aim of our study was to investigate the influence of an oral immunoenhanced supplement (arginine and glutamine) on nutritional and biochemical parameters in postsurgical ambulatory patients with head and neck tumor. DESIGN: A population of 39 ambulatory postsurgical patients with oral and laryngeal cancer was enrolled. At Hospital discharge postsurgical head and neck cancer patients were asked to consume two units per day of a specially designed enhanced supplement for a twelve week period. RESULTS: The mean age was 60.2+/-13.1 years (9 female/30 males). Duration of supplementation was 90.8+20 days. A significant increase of albumin (3.1±0.6 g/dl vs 4.12+0.7 g /dl; p<0.05), prealbumin (21.4±6.3 mg/dl vs 22.4+5.9 mg/dl; p<0.05) and transferrin (198.8±45.2 mg/dl vs 253.8+60.7 mg/dl; p<0.05) levels were observed. No differences were detected in weight and other anthropometric parameters. Ten patients (41.3%) received radiotherapy along the enhanced supplementation period and only 5 (20% of patients with radiotherapy) developed a clinical oral mucositis. CONCLUSIONS: At dose used, arginine and glutamine enhanced formula improved seric protein levels in ambulatory postoperative head and neck cancer patients with a low rate of oral mucositis in the subgroup with radiotherapy.

Guidelines to Subcontracting Clinical Laboratory Examinations a Proposal of the Catalan Association for Clinical Laboratory Sciences.

Clinical laboratories, acting as subcontractors, do not always supply information on the quality of their examination procedures and results. Thus, its selection by a contracting organization is often based only in economical criteria. This article gives some guidelines to subcontractors on how to describe its quality characteristics in order to be appropriately contracted.