Serotype-independent detection of foot-and-mouth disease virus

Foot-and-mouth disease virus (FMDV) causes a highly contagious vesicular disease affecting cloven hoofed animals and is considered the most economically important disease worldwide. Recent FMD outbreaks in Europe and Taiwan and the associated need for rapid diagnostic turnaround have identified limitations that exist in current diagnostic capabilities. To aid improved diagnosis, a serotype-independent FMDV antigen capture assay was developed using antibodies directed against a highly conserved cross-reactive protein fragment (1AB') located within the structural protein 1AB. Cattle sera raised against all 7 serotypes of FMDV bound purified 1AB' demonstrating its immunogenicity in infected animals. Polyclonal anti-1AB' antiserum was produced in chickens and applied as a universal detector of FMDV antigen. Western blot analysis and ELISA both demonstrated that anti-1AB' serum could recognize FMDV antigens independent of serotype. Two recently characterized anti-FMDV monoclonal antibodies were also evaluated for their ability to capture FMDV antigen independently of serotype. When used in combination with chicken anti-1AB' antibodies in an antigen capture ELISA format, all serotypes of FMDV were detected. These data represent the first demonstration of the use of serotype-independent FMDV antigen capture reagents which may enable the development of rapid laboratory based assays or perhaps more significantly, rapid field-based pen-side or point of entry border control diagnostic tests.     

Development and application of monoclonal antibodies against avian influenza virus nucleoprotein

Rapid and accurate diagnosis of avian influenza (AI) infection is important for an understanding epidemiology. In order to develop rapid tests for AI antigen and antibody detection, two monoclonal antibodies (mAbs) against influenza nucleoprotein (NP) were produced. These mAbs are designated as F26-9 and F28-73 and able to recognize whole AI virus particles as well as the recombinant NP. Both of the mAbs were tested in a slot blot for their reactivity against 15 subtypes of influenza virus; F28-73 reacted with all tested 15 subtypes, while F26-9 failed to react with H13N6 and H15N8. The mAb binding epitopes were identified using truncated NP recombinant proteins and peptide array techniques. The mAb F26-9 reacted with NP-full, NP-1 (638bp), NP-2 (315bp), NP-4 (488bp), and NP-5 (400bp) in the Western blot. The peptide array results demonstrated that the mAb F26-9 reacted with NP peptides 15-17 corresponding to amino acids 71-96. The mAb F28-73 recognized the NP-full, -1 and -4 fragments, but failed bind to NP-2, -3, -5, and any peptides. This antibody-binding site is expected to be contained within 1-162 amino acids of AI NP, although the exact binding epitope could not be determined. The two mAbs showed reactivity with AI antigen in immunofluorescence, immunohistochemistry and immune plaque assays. Immune response of AI infected animals was determined using the mAb F28-73 in a cELISA. All tested chickens were positive at 11 days post-infection and remained positive until the end of the experiment on day 28 (>50% inhibition). The two mAbs with different specificities are appropriate for developing various tests for diagnosis of AI infection.     

Development and evaluation of an IgM-capture ELISA for detection of recent infection with bluetongue viruses in cattle

An IgM-capture enzyme-linked immunosorbent assay (ELISA) was developed for the detection of recent infection of bluetongue virus (BTV) in cattle. The test is based on the use of biotinylated capture anti-bovine IgM antibodies bound to a streptavidin-coated ELISA plate. The captured IgM antibodies were detected by application of BTV VP7 antigen and a VP7 antigen-specific monoclonal antibody. The IgM-capture ELISA was compared with the competitive ELISA by testing serum samples from groups of calves infected experimentally with five USA and 19 South Africa serotypes of BTV. The IgM-capture ELISA was able to detect bovine anti-VP7 antibodies from all animals infected with the 24 BTV serotypes at 10 days post-infection, whereas the competitive ELISA was not. When the detectable IgM diminished after 40 days post-infection by the IgM-capture ELISA, the IgG anti-VP7 antibodies remained high. The IgM-capture ELISA is sensitive and can be applied for the detection of recent infection of BTV in cattle.     

HLA-Cw*1701 is associated with two sub-Saharan African-derived HLA haplotypes: HLA-B*4201, DRB1*03 and HLA-B*4202 without DRB1*03

Different extended haplotypes have been described for many ethnic groups, such as African-Americans. The complotype FC(1,90)0 is in linkage disequilibrium with HLA-B42, DRB1*0302 in African-Americans and Southern African Xhosa individuals, suggesting a common ancestry. In order to analyze the distribution of Cw*17 alleles (Cw*1701, 1702) in relation to this African-derived extended haplotype, we studied a large panel of samples from African-American individuals and additionally a group of selected samples carrying HLA-B42, DR3 and HLA-B42, non-DR3 antigens. HLA alleles were assigned using sequence-specific amplification (SSP) and sequence-specific oligonucleotide probe hybridization (SSOP). We have found that all haplotypes (10 in total) carrying the extended haplotypes [HLA-B42, FC(1,90)0, DRB1*0302] were positive for HLA-Cw*1701. Interestingly, HLA B*4201 was found in all samples (17 in total) carrying HLA-B42, DR3, Cw*1701, whereas HLA-B*4202 was found in 10 out of 13 samples from individuals carrying HLA B42, Cw*1701 non-DR3. These findings suggest that HLA-Cw*17 polymorphism is conserved in different ethnic populations and that HLA-B42 alleles seem to separate at least different African-derived haplotypes. The historical context of these findings are important for the study of human evolution and they may be useful for the development of strategies in the search for possible donors in organ transplantation for African-derived populations.     

Robotic extended pyelolithotomy for complete staghorn calculus

Staghorn stones represent a therapeutic challenge to urologists. We present our experience with laparoscopic extended pyelolithotomy for treatment of staghorn and complex renal calculi in highly selected cases. This approach provides the principles of open surgery with the advantages of minimally invasive surgery. We describe our experience with robot-assisted extended pyelolithotomy for complex coralliform calculi. Since January 2007, robotic extended pyelolithotomy has been performed by transperitoneal approach in two patients with complete coralliform lithiasis (calculi average size 8 cm). One patient had history of percutaneous nephrolithotomy. Demographic and operative data were collected. All procedures were technically successful without need for open conversion. Mean estimated blood loss was 175 ml (range 50–300 ml), and mean operative time was 150 min (range 120–150 min). A perinephric drain was employed in one patient with duration of 5 days. Postoperative imaging confirmed complete stone clearance. Robotic extended pyelolithotomy is a feasible and reproducible procedure for removal of complete and partial staghorn calculi in selected patients with complex nephrolithiasis. This approach might limit the role of open surgery for these calculi, but further publications with more cases are necessary to further define its utility.     

Geographical Information Systems: A Tool to Map and Analyze Disease Spread

Objective

Use GIS to illustrate and understand the association between environmental factors and spread of infectious diseases.

Introduction

Spatial methods are an important component of epidemiological research motivated by a strong correlation between disease spread and ecological factors (1). Our case studies examine the relationship between environmental conditions, such as climate and location, and vector distribution and abundance. Therefore, GIS can be used as a platform for integrating local environmental and meteorological variables into the analysis of disease spread, which would help in surveillance and decision making.

Methods

Case study 1- Lyme disease -Lyme disease is a tickborne infection caused by the bacterium Borrelia burgdorferi. The goal of this study was to analyze the association between meteorological factors and Lyme disease risk in humans in Texas. A total of 1,212 cases reported from 138 counties in Texas from Jan 2000 to Dec 2010 were analyzed. We used temperature and precipitation raster grids to generate humidity maps for Texas region. Our results indicated that there is a strong positive association between Lyme disease incidence and humidity, with western cross timbers region having a higher risk then the low plains.Case study 2- Spinach – Motivated by the recent increase in food-borne outbreaks related to fresh produce, one of the objectives for this study was to use the geospatial analysis to elucidate factors that contribute to contamination of produce at preharvest. We collected 955 spinach samples from 13 produce farms in Colorado and Texas during 2010–2011 and tested the samples for Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella contamination. The spinach contamination results were then used in conjunction with the National Resource Information (NRI) databases along with the SSURGO database to predict environmental and meteorological factors contributing to spinach contamination. Our findings would help to reduce frequency of human foodborne illnesses related to fresh produce.Case study 3- Valley fever Coccidioidomycosis or Valley Fever (VF) is a fungal zoonosis affecting humans and a variety of animal species. In this study, we used Texas Veterinary Medical Diagnostic Laboratories (TVMDL) data of all dog sera tested for coccidioidomycosis from July 1999 - December 2009. Census data on human population density for Texas were used to determine the dog population density and identify disease clusters for the 5,871 submitted dog sera over a period of 10.5 years. Both the isopleth map of the VF seropositive rates in dogs across Texas and the identified spatial and spatio-temporal clusters of the disease suggested that VF occurs in the western and southwestern part of Texas at a much higher rate than in other areas of the state (2). Since VF is not a reportable disease in TX, dogs could be used as a sentinel for human infection.

Results

The above studies illustrate the utility of GIS as a tool in integrating different ecological factors to understand disease occurrence and spread. The geographical and temporal patterns found in these studies provide benchmark to support disease control activities in Texas. Additionally, the identification of high-risk areas may be useful for decision makers to improve and prevent future disease spread.

Conclusions

Spatial epidemiological research has challenges, such as dealing with coarse level and aspatial datasets. Testing laboratories provide limited spatial information up to the zip code level due to the confidentiality concerns. Spatial analysis of such dataset prevents research at finer resolutions (Census block or block group). Despite these limitations, spatial epidemiology continues to be an invaluable field in the research and surveillance of infectious disease.     

Single port radical prostatectomy: current status

The aim of this study is to analyze the current literature on single port radical prostatectomy (LESS-RP). Single port radical prostatectomy laparoendoscopic (LESS-RP) has established itself as a challenge for urological community, starting with the proposal of different approaches: extraperitoneal, transperitoneal and transvesical, initially described for laparoscopy and then laparoscopy robot-assisted. In order to improve the LESS-RP, new instruments, optical devices, trocars and retraction mechanisms have been developed. Advantages and disadvantages of LESS-RP are controversial, while some claim that it is a non-trustable approach, regarding the low cases number and technical difficulties, others acclaim that despite this facts some advantages have been shown and that previous described difficulties are being overcome, proving this is novel proposal of robotics platform, the Da Vinci SP, integrating the system into “Y”. The LESS-RP approach gives us a new horizon and opens the door for rapid standardization of this technique. The few studies and short series available can be result of a low interest in the application of LESS-RP in prostate, probably because of the technical complexity that it requires. The new robotic platform, the da Vinci SP, shows that it is clear that the long awaited evolution of robotic technologies for laparoscopy has begun, and we must not lose this momentum.     

Loss of BAP1 expression in metastatic tumor tissue is an event of poor prognosis in patients with metastatic clear cell renal cell carcinoma

Purpose

To evaluate the prognostic impact of the protein expression of both PBRM1 and BAP1 in metastatic tissue of patients with metastatic clear cell renal cell carcinoma (ccRCC).