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Biomedical subjects

Jose Garcia

Publications and source records attributed to Jose Garcia.

8 recordsLinked to original sources

Infectious pancreatic necrosis virus suppresses type I interferon signalling in rainbow trout gonad cell line but not in Atlantic salmon macrophages.

RTG-P1 cells are a rainbow trout fibroblastic cell line permanently transfected with the luciferase gene under the control of the Mx promoter. On exposure to interferon (IFN) or IFN inducing agents, the cells produce luciferase. IPNV did not induce luciferase production up to 24h post-infection but did not suppress constitutive luciferase production. Furthermore, IPNV suppressed luciferase production induced by poly I:C. RT-PCR analysis of IPNV infected cells showed IFN gene transcription from 6h post-infection with increasing expression up to 24h. Housekeeping genes beta-actin and GAPDH were also expressed along with upregulation of IRF1 and slight upregulation of STAT1. When RTG-P1 cells were stimulated with IFN, Mx transcripts, measured by qRT-PCR, peaked at 3-6h and thereafter fell to low levels, but in the presence of IPNV, Mx transcription at this time was significantly suppressed but continued to rise gradually. Luciferase production was lower in infected cells at 12h post-infection but not significantly after 24h. These results indicate that, in non-stimulated RTG-P1 cells, while IPNV induces IFN transcription, activation of Mx expression is suppressed. Furthermore, when stimulated by IFN, the rate of Mx transcription is significantly suppressed by the virus. This would probably give time for the virus to replicate rapidly in the early phases of infection. Contrary to the fibroblastic cell line, IPNV stimulated IFN production by salmon macrophages in vitro at least as strongly as poly I:C, with no suppression of the IFN response to poly I:C, and the virus persisted for up to 9 days without causing CPE.

Animals↗

Expression and processing of hepatitis C virus structural proteins in Pichia pastoris yeast.

Development of heterologous systems to produce useful HCV vaccine candidates is an important part of HCV research. In this study different HCV structural region variants were designed to express the first 120 aa, 176 aa, 339 aa, and 650 aa of HCV polyprotein, and aa 384 to 521, or aa 384-605 or aa 384-746 of HCV E2 protein fused to the leader sequence of sucrose invertase 2 allowing the secretion of recombinant E2 proteins. Low expression levels were observed for HCV core protein (HCcAg) variants expressing the first 120 aa and 176 aa (HCcAg.120 and HCcAg.176, respectively). Higher expression levels were observed when HCcAg was expressed as a polypeptide with either E1 or E1 and E2 proteins. In addition, HCcAg was processed to produce two antigenic bands with 21 and 23kDa (P21 and P23, respectively) when expressed as a polypeptide with HCV E1 and E2 proteins. Results also suggest E1 processing in the context of HCcAg.E1.E2 polyprotein. On the other hand, E2.521, E2.605, and E2.680 were efficiently excreted to the culture medium. However, the entire E2.746 variant predominantly localized in the insoluble fraction of ruptured cells. Results suggest that the hydrophobic C-terminal E2 region from aa 681 to 746 is critical for intracellular retention of recombinant E2.746 protein in Pichia pastoris cells. Endo H or PNGase F treatment suggests that E2.746 was modified with high-mannose type oligosaccharides in P. pastoris. These data justify the usefulness of P. pastoris expression system to express HCV structural viral proteins which may be useful targets for HCV vaccine candidates.

Antigens, Viral↗

Distal hamstring lengthening in cerebral palsy: the influence of the proximal aponeurotic band of the semimembranosus.

We undertook a prospective review of 24 children with spastic diplegia treated by distal hamstring fractional lengthening at the Sheffield Children's Hospital. In 14 children (24 limbs) the correction achieved (popliteal angle) was inadequate and in these patients in the operating room a further correction was attained by dividing a tight band palpated in the substance of the semimembranosus muscle. Under general anaesthetic (preoperatively) the popliteal angle (a degrees ) was measured using a goniometer, then again (b degrees ) following distal hamstring fractional lengthening and finally (c degrees ) after surgically dividing the tight band. The mean preoperative popliteal angle (a degrees ) in all 24 limbs was 65 degrees (52-90). Following the standard hamstring fractional lengthening it (b degrees ) measured 37 degrees (35-50) and after division of the tight band it (c degrees ) measured 15 degrees (10-20). The reduction in popliteal angle following release of the tight band (proximal aponeurosis) was statistically significant (P<0.05). We undertook a cadaveric examination of 22 lower limbs and confirmed in all the cases that this band was the proximal aponeurosis of the semimembranosus muscle. The proximal aponeurosis is a well defined band located at the anterior aspect of the semimembranosus muscle where it arises from the tendon of the proximal attachment. It is separate from the distal aponeurosis. Division of the proximal aponeurosis during fractional lengthening of the distal hamstring in patients with cerebral palsy results in a significant reduction in the flexion deformity.

Adolescent↗

Free-floating left atrial ball thrombus following mitral valve replacement.

We report the case of a 51-year-old woman who underwent mitral valve replacement for prolapse with severe regurgitation, depressed ejection fraction, and atrial fibrillation. Two weeks post-operatively, a transesophageal echocardiogram was performed for bacteremia. The patient was found incidentally to have a large free-floating ball thrombus in the left atrium. The patient was managed with anticoagulation because of the high-risk nature of repeat surgery. One month following diagnosis, the patient still had persistent thrombus in the left atrium seen on transthoracic echocardiography despite therapeutic anticoagulation. Free-floating ball thrombus is a rare and dramatic finding seen on echocardiography in patients with mitral valve disease.

Echocardiography↗

Easy surgical approach for completion of biventricular pacing.

Placement of a pacing lead into a branch of the coronary sinus for biventricular pacing sometimes is difficult or impossible. Surgical completion typically has included immediate or subsequent thoracotomy lead placement with hookup to the device at the time of chest surgery. We describe an alternative procedure of complete device-lead hookup and permanent pocket closure in the electrophysiology laboratory. The left ventricular lead is an epicardial type. The lead is tunneled to a position where the surgeon subsequently can recover it using the thoracotomy incision and implant the lead on the epicardium using device-based testing.

Aged↗

Randomized, controlled trial of slow versus rapid feeding volume advancement in preterm infants.

OBJECTIVES: To determine whether infants who are fed initially and advanced at 30 mL/kg per day (intervention) take fewer days to get to full feedings than those who are fed initially and advanced at 20 mL/kg per day (control), without increasing their incidence of feeding complications and necrotizing enterocolitis (NEC). We also examined whether these infants regain birth weight earlier, have fewer days of intravenous fluids, and a have shorter hospital stay. METHODS: A randomized, controlled, single-center trial was conducted in a Neonatal Intensive Care Unit of a community-based county hospital in Houston, Texas. Infants between 1000 and 2000 g at birth, gestational age < or =35 weeks, and weight appropriate for gestational age were allocated randomly to feedings of expressed human milk or Enfamil formula starting and advanced at either 30 mL/kg per day or 20 mL/kg per day. Infants remained in the study until discharge or development of stage > or =IIA NEC. RESULTS: A total of 155 infants were enrolled: 72 infants in the intervention group and 83 in the control group. Infants in the intervention group achieved full-volume feedings sooner (7 vs 10 days, median), regained birth weight faster (11 vs 13 days, median), and had fewer days of intravenous fluids (6 vs 8 days, median). Three infants in the intervention group and 2 control infants developed NEC for an overall incidence of 3.2% (relative risk: 1.73; 95% confidence interval: 0.30-10.06). CONCLUSION: Among infants between 1000 and 2000 g at birth, starting and advancing feedings at 30 mL/kg per day seems to be a safe practice and results in fewer days to reach full-volume feedings than using 20 mL/kg per day. This intervention also leads to faster weight gain and fewer days of intravenous fluids.

Enteral Nutrition↗

Synchronous multiple glomus tumors of the esophagus and lung.

A case of synchronous esophageal and multiple pulmonary tumors presenting complex diagnostic problems is described. In the course of routine pulmonary screening a pulmonary coin lesion of the right lung and three very small foci on the left side, under the pleura falling just within the range of diagnostic parameters was identified in a symptom-free patient. In addition to the multiple lung lesions a single sub-mucosal esophageal tumor was detected. Following minimally invasive surgical excision of the tumors the precise nature of the neoplasms was determined by means of comparative histological, light-and electro-microscopic as well as immunohistochemical studies. The earlier diagnosis of carcinoid was reviewed, rejected and glomus tumor was confirmed. Multiple glomus tumors of the above localizations have not been previously described in the literature.

Adult↗