Development of nested PCR assays for detection of bovine respiratory syncytial virus in clinical samples

Two nested PCR assays were developed for the detection of bovine respiratory syncytial virus (BRSV). Primers were selected from the gene encoding the F fusion protein (PCR-F) and the gene encoding the G attachment protein (PCR-G). Biotinylated oligonucleotide probes, termed F and G, were selected for the hybridization of the respective PCR products. The sensitivities of the PCR-F and PCR-G assays were similar, both detecting 0.1 tissue culture infective dose of the virus. The PCR-F assay amplified all bovine strains and one human strain (RS32) tested. No cross-reactions were observed with nine heterologous respiratory viruses. PCR-F products of bovine and human RSV strains were discriminated by using endonuclease restriction enzyme ScaI, which specifically cleaved, products of BRSV. Oligonucleotide probe F was also specific for products of BRSV. The PCR-G assay detected all bovine strains and none of the human strains tested. A faint electrophoretic band was also observed with products of Sendai virus. However, probe G did not hybridize with this product, only with products of BRSV. Nasal swabs collected from cattle with no symptoms and cattle in the acute stage of respiratory disease were analyzed for BRSV by the immunofluorescence (IF) method and by the PCR-F and PCR-G assays. The virus was detected by the PCR assays in 31 of 35 (89%) samples tested. Only 23 samples (66%) were positive by the IF method, and these samples were also positive by both the PCR-F and PCR-G assays. The 31 samples detected as positive by PCR originated from cattle presenting clinical signs of acute respiratory disease; the four PCR-negative samples originated from clinically asymptomatic neighboring cattle. All sampled animals subsequently seroconverted and became reactive to BRSV. Thus, the detection of BRSV by PCR correlated with clinical observations and was considerably more sensitive (66 versus 89%) than IF. These results indicate that both nested PCR assays provide rapid and sensitive means for the detection of BRSV infection in cattle. Considering its higher specificity, the PCR-F assay can be recommended as the method of choice in the analysis of clinical specimens of BRSV.     

Nitric oxide synthase activity in tissues of the blowfly Chrysomya megacephala (Fabricius, 1794).

Although insects lack the adaptive immune response of the mammalians, they manifest effective innate immune responses, which include both cellular and humoral components. Cellular responses are mediated by hemocytes, and humoral responses include the activation of proteolytic cascades that initiate many events, including NO production. In mammals, nitric oxide synthases (NOSs) are also present in the endothelium, the brain, the adrenal glands, and the platelets. Studies on the distribution of NO-producing systems in invertebrates have revealed functional similarities between NOS in this group and vertebrates. We attempted to localize NOS activity in tissues of na?ve (UIL), yeast-injected (YIL), and saline-injected (SIL) larvae of the blowfly Chrysomya megacephala, using the NADPH diaphorase technique. Our findings revealed similar levels of NOS activity in muscle, fat body, Malpighian tubule, gut, and brain, suggesting that NO synthesis may not be involved in the immune response of these larval systems. These results were compared to many studies that recorded the involvement of NO in various physiological functions of insects.     

Percutaneous transluminal angioplasty of the renal artery in transplanted kidneys

The objective of the work was to evaluate the contribution of percutaneous transluminal angioplasty (PTA) to treatment of hypertension and the graft function of a transplanted kidney. Angioplasty of the graft artery was performed in 39 patients. Technical success rate: 85%. Complications: 13%--in particular during the first years after introduction of the method. Twenty seven patients were followed up on a long-term basis. Changes of the median arterial pressure (MAP) were evaluated as well as glomerular filtration (GF) assessed by creatinine clearance. Mean values: [table: see text] A statistically significant (p < 0.01) reduction of the median arterial pressure (drop by > 10 mm Hg) along with reduction of the number of antihypertensive drugs was recorded in 70% of the patients during the first year and in 80% kidney recipients in investigations extending beyond one year. A significant increase of glomerular filtration (by > 20%) was found in 40% of the patients, on average the rise was insignificant. During the follow up period 13 kidney recipients were transferred to a dialyzation programme due to failure of the grafts as a result of rejection. CONCLUSION: Percutaneous transluminal angioplasty of a stenosis of the graft artery is the method of first choice in the treatment of hypertension. It is of fundamental importance also in rare cases in order to preserve or improve graft function and thus delay dialyzation treatment.     

Microprocessor controlled limitation system for a stand-alone freely movable treatment couch

Because of the capability of free movement in the treatment room, we recently introduced a Hercules treatment couch on one of our linear accelerators. One of the advantages of this couch is that it allows for a more flexible way of patient setup and that it can be moved entirely out of the way to enable treatment with a hospital bed. A disadvantage, however, is that the couch can hit a wall or a cover of the accelerator accidentally. A limitation system has been developed to protect both the table and the accelerator against such collisions.     

Implementation of a New Linear Control Technique Based on Experimentally Validated Small-Signal Model of Three-Phase Three-Level Boost-Type Vienna Rectifier

In this paper, the design and implementation of a new multiple-input-multiple-output linear control technique based on a theoretically established and experimentally validated small- signal model for the three-phase three-level boost-type ac/dc Vienna converter are presented. Averaging and local linearization techniques are used to derive the dynamic model expressed in the dqo reference frame. The resulted transfer functions are discretized for the sake of a digital controller design. Multiple-loop control strategy is adopted and consists of inner current feedback loops, which are based on the straightforward looping technique that neglects interactions between the dq components of control inputs and currents, respectively, and of an outer voltage loop, which is designed to ensure dc voltage regulation by adjusting the magnitude of the references for the inner current loops. The output dc voltage unbalance is also controlled in the inner loops. The proposed modeling and control approaches are first simulated and then validated on a 1.5-kW laboratory prototype supported by the DS 1104 digital real-time controller board of dSPACE. The obtained results prove the accuracy of the proposed new small-signal model and, therefore, its reliability for dynamic analysis and control design purposes. It is also proved that a judicious choice of controller parameters, as well as an adequate rating of boost inductors, allows one to meet the IEEE standard requirements in terms of ac line-current total harmonic distortion and power factor. The efficiency of the proposed control technique is maintained in case of disturbances occurring on both source and load sides.     

Proteolytic regulation of the extent of dietary proteins with skin grape proanthocyanidin and anthocyanidin's interactions

During technological processing, proanthocyanidins and anthocyanidins could be partly lost due to the complexation phenomena, affecting food and beverage nutritional properties, organoleptic properties and health-promoting potentials. A common issue is encountered when processing food and beverage which is binding of phenolics to dietary proteins. The present investigation aims at evaluating the proteolysis contribution, using pure protease (Pepsin, 3000 units g⁻¹), to protein–anthocyanidin and protein–proanthocyanidin interactions. Bovine serum albumin (BSA), ovalbumin (OVA) and casein (CAS) dietary protein models were used. High-performance liquid chromatography coupled to diode array detector (HPLC-DAD) and size exclusion chromatography analyses proved that pepsin treatment significantly (P > 0.05) decreased the ratio of flavonoids’ interaction with tested proteins . The proteolysis reduced anthocyanidin interactions with CAS, OVA and BSA by 64.88%, 57.37% and 42.87% respectively. Similarly, proanthocyanidins interaction with CAS, OVA and BSA were reduced by 34.23%, 13.74% and 2.39% respectively. This study provides the basis to develop innovative technologies to limit protein–flavonoid complexation during food and beverage processing.