Development and Validation of a Fluorescent Microsphere Immunoassay for Soluble CD30 Testing

Testing for soluble CD30 (sCD30), an indicator of Th2 immune response, is a useful prognostic marker in solid organ transplantation, lymphoproliferative disorders, autoimmunity, and various parasitic diseases. In this study we report the development and validation of a fluorescent microsphere immunoassay for the detection of sCD30 in serum, plasma, and culture supernatants. The dynamic range of this assay is 1 to 400 ng/ml, and the rate of recovery of various concentrations of recombinant sCD30 ranges from 97 to 116% (average recovery, 105%). The test showed a high degree of precision in both intra-assay and interassay studies (coefficients of variation, as high as 7% and 8%, respectively), with a sensitivity of 1 ng/ml. The normal reference range calculated for a cohort of 151 healthy individuals was 1 to 29 ng/ml. The clinical usefulness of the sCD30 fluorescent microsphere immunoassay was demonstrated by showing that levels of sCD30 have a positive correlation with specimens containing high titers of anti-double-stranded DNA antibodies and high titers of immunoglobulin G against Leishmania species. Given the multiplexing potential of the sCD30 fluorescent microsphere immunoassay reported in this study, it is expected that testing of sCD30 concentrations along with those of other cytokines will become an important diagnostic tool for selected immunological and inflammatory diseases where Th2-type cytokine responses have been reported.CD30 (TNFRSF 8) is a transmembrane protein, a member of the tumor necrosis factor (TNF) receptor superfamily. It was originally described as a marker for Reed-Sternberg cells (“Ki-1 antigen”) in Hodgkin''s disease (12, 18, 20). CD30 is expressed on CD4⁺ and CD8⁺ T cells that secrete Th-2 type cytokines (8, 17). Signaling through CD30 plays important roles in T- and B-cell growth, differentiation, and function. The soluble form of CD30 (sCD30) is produced after proteolytic cleavage of the membrane-bound CD30 ectodomain by the TNF-α-converting enzyme (9).Numerous studies have reported that circulating levels of sCD30 may represent a biomarker for outcomes in solid-organ transplantation (16, 21). In addition, other studies have reported that levels of sCD30 have important prognostic value for various lymphoproliferative disorders (4, 15, 22), systemic lupus erythematosus (SLE) (5, 7), and leishmaniasis (1, 2). The current method for quantitation of sCD30 is the enzyme-linked immunosorbent assay (ELISA), which has good sensitivity and specificity. However sCD30 production differs greatly between patients, and the dynamic range of ELISAs requires that many samples be diluted and retested. Moreover, ELISA measures only 1 analyte per well, which precludes the testing of multiple analytes in the same test. In this study, we report the development and validation of a fluorescent microsphere immunoassay suitable for multiplexed determination of sCD30 levels, along with those of other cytokines, in serum and plasma specimens and in tissue culture supernatants. We present data showing the positive correlation of sCD30 levels with titers of anti-double-stranded DNA (anti-dsDNA) antibodies in SLE and with immunoglobulin G (IgG) levels in patients with leishmaniasis.     

Expression of Growth Factors in Endotheliocytes in Vascular Malformations

The expression of growth factors and proliferation of endotheliocytes in vascular malformations were studied by immunohistochemical methods. The detected specific features of growth factor expression in the endothelium of venous and arteriovenous malformations seem to reflect the differences in the pathogenesis of these formations. High proliferative activity of the endothelium in angiodysplasias of both types can underlie the disease relapsing. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 147, No. 3, pp. 341–345, March, 2009     

Removal of tracheobronchial foreign bodies in adults using flexible bronchoscopy: experience 1995–2006

Background  Tracheobronchial foreign body (TFB) removal in adult patients using the combined technique of flexible bronchoscopy (FBSC) through the endotracheal tube as well as using the simple FBSC has not often been described. This study reports our experience with FBSC for removal of TFBs and describes diagnosis, techniques of removal, and types of TFBs. Methods  We retrospectively examined bronchoscopic records of adult patients performed between 1995 and 2006 and collection of foreign bodies in Clinical Department for Respiratory Diseases, University Hospital Split, Croatia. Results  TFBs were found in 86 (0.33%) out of 26,124 patients who were submitted to bronchoscopy. The majority of the patients (90%) had some risk factor for aspiration, among which stroke (30%) was the most frequent. Patients with different neurologic and neuromuscular diseases together accounted for 58% of all patients with TFB aspiration. Medical history was suggestive of foreign body aspiration in 38.4% of the patients, while chest X-ray was indicative in 7% of the patients. TFBs were most often found in the right bronchial tree (75.6%). The most common TFBs were animal and fish bones (39.5%). In 90.7% of the patients they were successfully removed under FBSC, whereas in 8.1% of the patients a TFB was extracted with flexible bronchoscope through endotracheal tube. Surgery was needed in only one case. Conclusions  Although foreign bodies in the tracheobronchial tree are rare in adults, the clinician must be aware of their likelihood. Foreign body aspiration should be considered especially in the etiology of recurrent lung diseases and in the presence of risk factors for aspiration, in particular with different neurologic and neuromuscular diseases. They can be successfully and safely removed in the majority of patients under local anesthesia by using FBSC. In cases when repeated procedure is needed, endotracheal tube is recommended.     

Transurethral ureteropyeloscopy with lithotripsy and lithoextraction in children

High efficacy of endoscopic treatment of urolithiasis in adults stimulated its introduction in the strategy of urolithiasis management in children. 133 ureteropyeloscopies with ureterolithotripsy and ureterolithoextraction was performed in 121 children (boys and girls, age 11 months to 16 years). Concrements (0.5-2.0 cm and larger) located in the caliceal-pelvic segment, ureter, 12 patients had bilateral urolithiasis. Ineffective extracorporeal lithotripsy (ELT) was followed by ureteropyeloscopy in 86 (71%) cases. Rigid and fibroureteropyeloscopes were used. Due to ureterolithotripsy and lithoextraction, concrements were removed completely in 98 (80.9%) patients including 5 children with ligature concrements and 11 children with ureteral strictures which required additional endoureterotomy and removal of ligature. In 23 (18%) cases when migration of concrements and their fragments did not allow complete elimination of the concrements, the kidney in ureteropyeloscopy was drained by catheters or stents with further ELT. Postoperative exacerbation of pyelonephritis treated conservatively was registered in 20 (16.5%) children. Long-term complications were not seen. Thus, transurethral uretheropyeloscopy with lithotripsy and lithoextraction is highly effective, low invasive, safe treatment of urolithiasis in children in failure of ELT.     

Effect of different schemes of general anaesthesia on dynamic of intellectual capacity restoration in patients operated for acute hemorrhages with high operative risk

The analysis of speed of the cognitive functions restoration, depending on the general anesthesia scheme applied was conducted. The impact of various schemes of the intensive therapy applied on the dynamics of index was investigated. Interrelationship between the homeostasis indices and the intelectual capacities patients, operated on for an acute bleeding in high operative risk, was established.