Small bowel obstruction secondary to crab shell bezoar: A case report.

Small bowel obstruction is a common surgical condition that needs surgical intervention if conservative measures fail. Bezoar is a rare aetiology of small bowel obstruction with incidence of 4.5%. The bezoars can be grouped, according to the content, into four common types: phytobezoars, trichobezoars, pharmacobezoars, and lactobezoars. However, unusual bezoars like plastic bezoars and metal bezoars have been reported too. Herein, we report a case of an elderly lady who was treated for small bowel obstruction due to crab shell bezoar. This is the first case reported in literature. Ingestion of large intact pieces of crab shell should be avoided due to the potential of causing small bowel obstruction.

Elevated plasma tissue factor levels in neonates with umbilical arterial catheter-associated thrombosis.

Catheterization of the umbilical artery has been a useful aid in the management of sick neonates for the past few decades. However, it is associated with various complications. Reported studies strongly suggest a significant role of intravascular catheterization in the development of aortic thrombi. Increase in thrombosis of large vessels is believed to be related to mechanical injury in the catheterized vessels, which provide direct exposure of blood to tissue factor (TF), the primary cellular initiator of the extrinsic coagulation pathway. This study was conducted to determine the levels of plasma TF, tissue factor pathway inhibitor (TFPI) and D-dimer (DD) in infants with umbilical arterial catheter (UAC)-associated thrombosis. Quantification of TF was carried out using an in-house sandwich ELISA, whereas TFPI and DD levels were measured with commercial immunoassay kits. Infants with UAC inserted were found to have significantly higher levels of plasma TF (p < 0.001) than baseline levels. However, there were no significantly elevated levels of TFPI or DD. Infants with UAC-associated thrombosis demonstrated a greater increase of TF level (median: 414.5 pg/mL; range: -76.0, 6667.0) than infants without UAC-associated thrombosis (105.0 pg/mL; -976.0, 9480.0; p = 0.009) following UAC insertion. Our findings indicate that quantification and monitoring of TF levels could predict thrombus formation in infants with indwelling UAC. Following umbilical arterial catheterisation, infants with an approximately 3-fold rise in plasma TF levels were most at risk of developing abdominal aorta thrombosis as confirmed by real-time abdominal ultrasonography.

An in-house ELISA method for quantification of circulating tissue factor.

Tissue Factor (TF) is a low molecular weight transmembrane glycoprotein that initiates the clotting protease cascade. It is considered to be the principal regulator of the extrinsic coagulation pathway, hemostasis and thrombosis, as well as inflammation and cellular immune response. An in-house two-step direct sandwich ELISA (enzyme-linked immunosorbent assay) for immunological quantification of plasma TF was successfully developed. The assay employed a monoclonal antibody against human TF (1:400 dilution; 1250 ng/ml) and peroxidase-conjugated anti-TF IgG (1:1000 dilution; 2000 ng/ml) as capture and detecting antibodies respectively, whilst tetramethylbenzidine/H2O2 were utilized as substrates. Titration curves of recombinant TF were linear within 10 to 4000 pg/ml, with a detection limit of 36.31 pg/ml. It demonstrated low intra- (2.50 - 9.23 CV%) and inter-assays (5.65 - 13.57 CV%) variability, as well as satisfactory analytical recovery (91.55 - 103.95%) and good parallelism. The assay developed was intended to be applied for measurement of plasma TF levels in patients with thrombotic disorders.

Comparative analysis of three permeabilization methods for cytofluorometric evaluation of cytoplasmic myeloperoxidase.

A comparative study was conducted to evaluate three different permeabilization methods: FACS Permeabilizing Solution (FPerm), CytoFix/CytoPerm Kit (CFP) and Paraformaldehyde-Tween 20 (PFT) reagents, in cytoplasmic labeling of myeloperoxidase (MPO). Peripheral blood cells from 23 healthy subjects were fixed and permeabilized according to the proposed procedures, prior to direct immunofluorescence staining with CD14, CD45, IgG1, IgG2 and MPO monoclonal antibodies (McAb). Subsequent flow cytometric analysis was performed on FACSCalibur flow cytometer (Becton Dickinson, BD). As far as the antigenic expression of MPO in normal samples is concerned, FPerm and CFP demonstrated better cytoplasmic staining by inducing minor effects on light-scattering properties of the cell populations, whereas PFT-treated samples showed a diminished ability to distinguish the cell types. However, the simple and rapid FPerm method required an earlier processing of samples since the stored whole blood samples (for more than 8 hours) tended to show a significant decrease of fluorescence intensity. We also have demonstrated that P/N ratio possesses added value in evaluation of cell reactivity in immunophenotyping, based upon the apparent nonspecific cytoplasmic staining of MPO in the lymphocyte population.

Flow cytometric analysis of intracellular myeloperoxidase distinguishes lymphocytes, monocytes and granulocytes.

A study was undertaken to evaluate the ability of flow cytometric analysis of intracellular myeloperoxidase (MPO) in differentiating populations of lymphocytes (L), monocytes (M) and granulocytes (G), by means of lysed whole blood method. Anticoagulated blood from 23 normal individuals was lysed with FACS lysing solution and permeabilized with FACS permeabilizing solution before subjected to direct immunofluorescence staining. The geometric means of the fluorescence intensity were measured using FACSCalibur flow cytometer (Becton Dickinson). Populations of L, M and G were gated based on their light scatter characteristics and expression of CD14 and CD45. Then, the fluorescence intensity of MPO expression was studied in these individual cell populations. The results showed that fluorescence intensity of MPO was the strongest in G and weakest in L, whereas M showed intermediate fluorescence intensity. Our findings reveal that discrimination of these three cell types is achievable based upon the sole expression of intracellular MPO.

Cloning, characterisation and expression of vitellogenin gene of Oreochromis aureus (Teleostei, Cichlidae).

cDNA synthesised from liver Poly A+ RNA of estradiol-stimulated male Oreochromis aureus was ligated to PTZ-19R and transformed into E. coli. TG-1 to yield 6 x 10(4) clones/ug DNA, of which 6% was vitellogenin positive. Restriction maps indicate 4 possible subgroups of vitellogenin cDNA, with homology, observed by Southern hybridisation, at a fragment flanked by Pst 1 sites. When the amino acid sequence derived from the DNA sequence of pOA Vg 62 was aligned with that of S. gairdneri, X. laevis, C. elegans and D. melanogaster, homologies of 63%, 22%, 21% and 15%, respectively, were obtained. Estradiol induced a vitellogenin gene transcript of 6500 nucleotides at 6 h and reached a maximum at 72 h after stimulation.