Simultaneous detection of antibodies to foot-and-mouth disease non-structural proteins 3ABC, 3D, 3A and 3B by a multiplexed Luminex assay to differentiate infected from vaccinated cattle

For the first time, a multiplex bead immunoassay was used to test simultaneously, with a single sample, the immune response to foot-and-mouth disease non-structural proteins 3ABC, 3A, 3B and 3D from experimentally infected and vaccinated cattle. We cloned and expressed these non-structural proteins (NSPs) as recombinant antigens. The purified proteins were coupled to microspheres labeled with anti-His monoclonal antibody with different proportions of red and orange fluorescent dyes and reacted against serum specimens. Antibody reacting against different NSPs, and thus, the different colored beads, was detected by use of the Luminex system. This multiplex bead immunoassay can detect the immune response to NSPs in cattle as early as 7 days post-infection. In general antibodies to the protein 3D appeared early after infection and anti-3ABC antibodies were detected at higher levels than the other NSPs. A clear differentiation was established between infected and vaccinated or uninfected cattle. The multiplex bead immunoassay was compared to individual indirect enzyme-linked immunosorbent assays (iELISAs) for the same NSP's responses. Results indicated that this new assay had a high positive correlation with those generated by iELISA. The Luminex-based technology promises to be a sensitive and efficient method that permits multiplexed NSP antibody detection from a single sample and would therefore provide both a time and cost saving to the laboratory.     

Diagnóstico prenatal de una trisomía parcial del cromosoma 1 (q25-qter) por translocación paterna

We report the prenatal diagnosis of a case of partial trisomy of chromosome 1 (q25-qter) due to paternal translocation in a 31-year-old patient. Amniocentesis for chromosomal analysis was performed in the 16th week of pregnancy because ultrasound examination had revealed certain anomalies. A necropsy of the foetus was carried out and the anomalies found were compared with those in other cases of partial trisomy of chromosome 1.     

Preoperative ultrasound‐guided fine‐needle aspiration cytology for axillary staging in breast carcinoma

Axillary lymph node (ALN) status is considered to be the single most important prognostic indicator in patients with breast cancer. It can be assessed by various radiological, pathological and surgical techniques, the most accurate being histological examination of lymph nodes after axillary lymph node dissection (ALND). This prospective study was conducted to assess the feasibility and diagnostic accuracy of preoperative ultrasound (US) and ultrasound‐guided fine‐needle aspiration cytology (USG‐FNAC) of ALN in patients with breast cancer. Thirty patients with FNAC‐proven breast cancer, planned for definitive surgery with axillary clearance, were included in this study. Ultrasonographic evaluation of the axillae of these patients was conducted for alterations in size, shape, contour and cortical morphology of lymph nodes that could reflect presence of underlying metastases. Ultrasound‐guided fine‐needle aspiration cytology of the ALN was done in 24 of these patients. These findings were evaluated, with the ALN status determined by histological examination after ALND. Out of the 30 patients, eight had T₁, 16 had T₂, five had T₃, and one had T₄ lesions. Ultrasound evaluation of the ALN had a sensitivity of 86.3%, a specificity of 41.6%, a positive predictive value of 79%, a negative predictive value of 50% and a diagnostic accuracy of 73.3%. Sensitivity of USG‐FNAC was 78.95%, specificity was 100%, positive predictive value was 100%, negative predictive value was 55.56% and diagnostic accuracy was 83.33%. Our study concludes that preoperative USG‐FNAC of ALN is a simple, minimally invasive, easily available and reliable technique for the initial determination of ALN status in patients with breast cancer. Those who are USG‐FNAC positive can be directed towards ALND straight away, and only those who are USG‐FNAC negative should be considered for sentinel lymph node biopsy. This will save considerable operating time, especially where facilities for sentinel lymph node biopsy (costly dye, gamma camera, nuclear medicine facilities) are restricted or not available.     

Evaluation of CrystalTM Rapid Stool/Enteric ID System for Identification of Aerobic Gram-negative Bacilli

Objective: The evaluation of the new miniaturized Crystal^TM Rapid Stool/Enteric System (Becton-Dickinson, USA) for identification of aerobic gram-negative bacilli.
Methods: a total of 154 clinical organisms ( Enterobacteriaceae: 120 strains; oxidase-positive fermenters: 13 strains; non-fermenters: 21 strains) were tested. Results were compared with those obtained with the PASCO^R system (Difco, USA) and divergent identifications were evaluated by standard biochemical tests.
Results: without additional testing, correct identification was obtained for 146 strains ( Enterobacteriaceae: 95%; oxidase-positive fermenters 87%; non-fermenters 100%). For adequate identification of Yersinia enterocolitica strains, however, panels had to be incubated for 5 instead of 3 hours.
Conclusions: the Crystal^TM Rapid Stool/Enteric system offers a safe, accurate and rapid method for the identification of frequent isolates of the family Enterobacteriaceae and bacterial stool pathogens.     

Comparison of a dipstick assay for detection of Brucella-specific immunoglobulin M antibodies with other tests for serodiagnosis of human brucellosis

A dipstick assay for the detection of Brucella-specific immunoglobulin M (IgM) antibodies was evaluated by studying the serological response of 133 cultures and or serologically confirmed patients with brucellosis in its different stages along with those of 34 healthy controls. As regards patients with illness less than 3 months in duration, 93.1% tested positive by the dipstick assay, a percentage similar to that obtained in the standard serum agglutination test (SAT) (92.0%), somewhat lower than that obtained by culture (100%) and higher than that obtained by IgM enzyme-linked immunosorbent assay (ELISA) (80.5%). SAT was the most sensitive test (87.0%) for patients with illness more than 3 months in duration, followed by culture (50%), the dipstick assay (28.3%), and IgM ELISA (7.5%). The results demonstrate that the dipstick assay could well be used in the serodiagnosis of patients with acute brucellosis, as well as to identify patients with a long history of the illness. Under laboratory conditions this test has the advantage of being quick and IgM antibody-specific.     

Development and use of a biotinylated 3ABC recombinant protein in a solid-phase competitive ELISA for the detection of antibodies against foot-and-mouth disease virus

A biotinylated 3ABC recombinant protein was developed and used in a competitive ELISA (cELISA) to detect foot-and-mouth disease virus (FMDV) antibodies in cattle, sheep and pigs. In this report, we describe the cloning and expression of 3ABC protein in Escherichia coli cells as fusion protein with 6xHis and biotin. This cELISA uses streptavidin to capture bacterially expressed and in vivo biotinylated 3ABC antigen. The antigen capture strategy provides a simple and reliable method, which does not require purification of recombinant antigen before the serological assay. An hyperimmune guinea pig antiserum produced against purified 6xHis-3ABC was used as competitor in the test.

The potential use of this cELISA for the identification of antibodies induced by FMD virus infection from those induced by vaccination is discussed.     

Children with intestinal failure as candidates for intestinal transplantation

Small bowel is not anymore considered a forbidden organ for transplantation, and intestinal transplantation (IT) is currently used as a therapeutic option in selected cases of permanent intestinal failure (PIF). Nevertheless, the experience is still scarce, and despite recent improvements, IT is nowadays only accepted as a life-saving option. However, small children are prone to suffer complications related to TPN, particularly end stage liver disease; moreover, suitable donor for the small baby who needs an IT is seldom available. Subsequently, a high pretransplantation mortality has been reported in the pediatric series. In those cases, the indication of IT shouldn't be delayed, and these children must be referred early for IT. Since we started our IT program, 17 children have been assessed, and 3 are currently on the waiting list; two for combined liver-small bowel transplantation (LSB), both with short bowel syndrome and end stage liver disease, and one patient with microuvillous dysplasia for isolated IT (indication loss of venous access). 3 children were referred too late and died, so did a fourth patient, candidate for LSB, before he could be transplanted.     

Pain relief: The perspective of catholic tradition

Efforts to study patient care from the perspective of Catholic ethics date back four centuries. In the course of this history, a prominent issue has always been management of pain and the efforts to avoid pain. Thus, Catholic theologians were concerned about the effects of pain medication upon the psychic function and considered whether or not hastening death for suffering people was allowed and the ethical norms for using opioids to remove pain when death is imminent. Moreover, the issue of “overmedication” for difficult or elderly patients has been a concern. The President's Commission on Ethics in Medicine and Human Research has utilized many of the principles developed by Catholic theologians when considering the matter of pain relief for dying persons.