Propagation of viruses infecting waterfowl on continuous cell lines of Muscovy duck (Cairina moschata) origin

Duck circovirus, duck hepatitis A virus 1, goose parvovirus and goose haemorrhagic polyomavirus are economically damaging pathogens of waterfowl, and replicate poorly or not at all in established cell lines. AGE1.CR, AGE1.CR.pIX and AGE1.CS cell lines, originating from the Muscovy duck, were tested for their suitability to isolate and identify these viruses. Immunofluorescence (IF) and quantitative polymerase chain reaction investigations verified that all cell lines are permissive for all four viruses; however, AGE1.CR.pIX proved to be the most productive and most sensitive for viral infection. IF experiments revealed that the time of one infectious cycle is approximately 12 to 14 h in the AGE1.CR.pIX cells in the case of the three DNA viruses, while it is 10 to 12 h for DHAV-1. Specific viral infectivity and the limit of detection by IF varied between 55 and 1484 copies, depending on the viruses and cell lines. Despite the high sensitivity of the cell lines for viruses, their viral productivity remained relatively low for the investigated field isolates. However, optimization of virus infection and/or the adaptation of the viruses to the cells can raise viral productivity and can make these cell lines suitable for vaccine development and production.     

Glutathione-dependent reduction of arsenate by glycogen phosphorylase responsiveness to endogenous and xenobiotic inhibitors.

Rabbit muscle glycogen phosphorylase-a (GPa) reduces arsenate (As(V)) to the more toxic arsenite (As(III)) in a glutathione (GSH)-dependent fashion. To determine whether reduction of As(V) by GPa is countered by compounds known to inhibit GP-catalyzed glycogenolysis, the effects of thiol reagents, endogenous compounds (glucose, ATP, ADP) as well as nonspecific glycogen phosphorylase inhibitors (GPIs; caffeine, quercetin, flavopiridol [FP]), and specific GPIs (1,4-dideoxy-1,4-imino-D-arabinitol [DAB], BAY U6751, CP320626) were tested on reduction of As(V) by rabbit muscle GPa in the presence of glycogen (substrate), AMP (activator), and GSH, and the As(III) formed from As(V) was quantified by high-performance liquid chromatography-hydride generation-atomic fluorescence spectrometry. The As(V)-reducing activity of GPa was moderately sensitive to thiol reagents. Glucose above 5mM and ADP or ATP at physiological levels diminished GPa-catalyzed As(V) reduction. All GPIs inhibited As(V) reduction by GPa in a concentration-dependent fashion; however, their effects were differentially affected by glucose (10mM) or AMP (200microM instead of 25microM), known modulators of the action of some GPIs on the GP-catalyzed glycogenolysis. Inhibition of As(V) reduction by DAB and quercetin was not influenced by glucose or AMP. Glucose that potentiates the inhibitory effects of caffeine, BAY U6751, and CP320626 on the glycogenolytic activity of GPa also enhanced the inhibitory effects of these GPIs on GPa-catalyzed As(V) reduction. AMP at high concentration alleviated the inhibition by BAY U6751 and CP320626 (whose antagonistic effect on GP-catalyzed glycogen breakdown is also AMP sensitive), whereas the inhibition in As(V) reduction by FP or caffeine was little affected by AMP. Thus, GPIs inhibit both the glycogenolytic and As(V)-reducing activities of GP, supporting that the latter is coupled to glycogenolysis. It was also shown that a GPa-rich extract of rat liver contained GSH-dependent As(V)-reducing activity that was inhibited by specific GPIs, suggesting that the liver-type GPa can also catalyze reduction of As(V).     

First results with wavefront-guided photorefractive keratectomy for hyperopia

PURPOSE: To compare the results of traditional laser photoablation and wavefront-supported customized ablation (WASCA) in hyperopic photorefractive keratectomy (H-PRK). METHODS: This was a prospective study, comparing two treatment groups, each comprising 40 eyes of 20 patients. Wavefront aberrations were examined using a Shack-Hartmann aberrometer. Preoperative refraction was similar in the two groups; in the traditional H-PRK group (Group 1) it was +3.10 +/- 0.85 D, and in the WASCA-guided group (Group 2) it was +2.90 +/- 0.80 D. H-PRK was performed with the Asclepion-Meditec MEL 70 flying-spot excimer laser. The follow-up time was 6 months. RESULTS: In Group 1, mean postoperative refraction was +0.14 +/- 0.24 D, and in Group 2, -0.10 +/- 0.25 D; mean uncorrected visual acuity was 0.92 +/- 0.16 in Group 1 and 0.95 +/- 0.18 in Group 2. Mean best spectacle-corrected visual acuity was 0.96 +/- 0.04 in Group 1 and 1.06 +/- 0.13 in Group 2. In Group 1, 67.5% (27 of 40 eyes), and in Group 2, 85% (34 of 40 eyes) were within +/-0.50 D of target refraction. Regarding change of spectacle-corrected visual acuity in Group 1, 5% (2 of 40 eyes), and in Group 2, 20% (8 of 40 eyes) gained one Snellen line compared to the preoperative; in Group 1, 10% (4 of 40 eyes), and in Group 2, 12.5% (5 of 40 eyes) lost two Snellen lines. In Group 2, the root mean square value for the higher order aberration increased from the initial 0.134 to 0.257 microm at 6 months after surgery. CONCLUSIONS: WASCA-guided hyperopic-PRK treatment was found to be safe and predictable. The results were better than those achieved with traditional PRK performed using the same flying-spot type excimer laser.     

Increased prevalence of platelet glycoprotein IIb/IIIa PLA2 allele in ischaemic stroke associated with large vessel pathology

Introduction: Platelet glycoprotein IIb/IIIa is a membrane receptor with a central function in the platelet adhesion and ultimately in the thrombus formation. Two major variants of the gene encoding the IIIa subunit, called PLA1 (A1) and PLA2 (A2), have been identified in the general population. There are indications that the A2 allele can also be associated with acute thrombosis or stroke. The purpose of this study was to study the distribution of the A2 allele in different vascular subtypes of stroke disease. Materials and methods: A total of 638 consecutive patients were analyzed and classified as having large vessel pathology (n=168) or a small vessel infarct (n=210). Localization of the vascular occlusions was deducted from analysis of the magnetic resonance imaging (MRI) scan results in stroke patients. The remainder patients were listed into a mixed vascular pathology group (n=167). Patients with other or poorly characterized stroke etiology were excluded from the study (n=93). Results: In the small vessel and mixed vascular pathology groups, the PLA2 allele frequency was similar to that in the controls. By contrast, PLA2 allele frequency was approximately two-fold higher in patients with large vessel pathology (23.3%) than in the stroke-free control subjects (11.7%, p<0.0005). Multivariate logistic regression analysis of data confirmed this association with an odds ratio (OR) of 2.9 (95% confidence interval [CI]: 1.6–4.9, p<0.0005). Conclusions: These data suggest that the PLA2 allele is more frequent in brain infarcts associated with large-vessel occlusion.     

Continuous postoperative epidural analgesia in abdominal surgery using ropivacain

We studied the selective block on patients receiving epidural Ropivacain (R) infusion for postoperative analgesia after major abdominal surgery. Twenty patients received R and twenty patients received Bupivacain (B) via peridural catheter during and after surgery. The patients' age ranged between 40 and 80 and they belonged to ASA I, II and III risk group. The epidural catheter was inserted one day before surgery and the proper position was tested by 80 mg Lidocaine. The epidural needle was inserted via T10-L1 interspaces in upper abdominal surgery and through L1-L3 interspaces in lower abdominal surgery. After the operation continuous epidural infusion of 2 mg/ml solution of R or 2.5 mg/ml solution of B was started. The infusion rate was changed according to the grade of sensory and motor block. The following parameters were observed during the postoperative 72 hours: blood pressure, heart rate, arterial blood O2 saturation, modified Bromage (BMG) score, verbal analogue scale (VAS), the spread of sensory block. Satisfactory sensory blockade was achieved with both local anaesthetics. The required daily dose of R and B increased during 72 hours. VAS scores reached their maximum level within 24 hours and were lower in the R group than in the B group but the difference was not significant. We experienced that 0.25% B causes more intense motor block than 0.2% R in equianalgetic dose but the difference did not reach a significant level. The infusion rate was often decreased because of the unwanted motor block caused by 0.25% B leading to insufficient postoperative analgesia. Because of this fact patients receiving B required opioid addition more often. Our conclusion is that R/B relative dose ratio is 1.2 suggesting that these local anaesthetics have different analgesic potency.     

Pancreas head resection combined with resection of the superior mesenteric vein: extensive procedure for tumors of the pancreas head

Surgical treatment for locally advanced cancer of the pancreas head with vascular invasion is controversial. It is regarded as a contraindication for resection by most surgeons, others perform resection involving the vessel. We performed pancreaticoduodenectomy with vascular resection in a 61 year old patient. The vascular reconstruction was performed using reversed saphenous vein. We review and analyse clinical, and operative data and results.     

Results of laparoscopic repair of ventral hernias--a case-control study

The surgical treatment of ventral hernias has changed in the last decade. Conventional methods involve tightening of the abdominal fascias is the reason for high recurrence rate. While the use of mesh reduces the recurrence rate, it does not change the rate of other complications, in particular, the risk of infection. We report the results of laparoscopic repair of ventral hernias performed in 15 patients (10 female, 5 male). The average age of patients was 59.5 years (39-79). Indications included 7 recurrent incisional hernias (3 patients had second recurrence, 4 patients had first recurrence operated on without mesh in the first operation); 5 patients with primary incisional hernias; and 3 primary umbilical hernias. The mean size of the defect was 66.2 cm2 (16-130 cm2). Average operation time was 101 minutes (64-190 min). In the postoperative period 1 patient developed seroma, and 4 patients developed ileus each was successfully treated conservatively. In one patient a second-look laparoscopy was performed because of neuralgia. Length of hospital stay varied between 3 and 10 days (median: 6 days); the length of follow-up period was 5-22 months (median: 12.4 months). We believe that laparoscopic treatment of ventral hernias reduces the complication rate, and reduces hospital stay.