Inflammatory bowel disease: diagnosis and research trends: the clinical lab is playing an increasingly important role.

IBD consists of ulcerative colitis and Crohn's disease and is characterized by recurrent and chronic inflammation of intestinal mucosa. In addition to patient's symptoms, history and physical examination, various techniques and tests help diagnose IBD. Endoscopy, histology, and imaging techniques such as CT-scan or MRI are used along with serological and hematological lab tests for its diagnosis. The likelihood of these procedures distinguishing between the diagnosis of ulcerative colitis or Crohn's disease is high. Various investigations are underway for better identification and management of IBD in the future.

Validation of the efficacy of a practical method for neutrophils isolation from peripheral blood.

OBJECTIVE: The objectives of this study were to validate the Polymorphprep method for neutrophils isolation for conditions within an identified clinical laboratory setting and to confirm the purity and viability of isolated neutrophils utilizing standard clinical laboratory cell counting, differential, and viability methods. DESIGN: A validation study, utilizing a quantitative and qualitative design, was performed. PATIENTS/SAMPLES: 18 EDTA tube blood samples were collected from donors between the ages of 18-60, with no underlying condition. The study was performed in a CLS student laboratory of a midwestern university. The Institutional Review Board approved the study and informed consent was obtained from each subject before blood collection. INTERVENTIONS: Blood samples were treated using a Polymorphprep procedure to isolate a pure population of neutrophils. MAIN OUTCOME MEASURES: Pre and post cell counts were performed using an automated cell counter. Manual white cells differentials and Trypan blue white blood cells viability analysis was performed. RESULTS: The mean purity levels were at least 94% following isolation with Polymorphprep, the mean viability of neutrophils was 99%. CONCLUSION: The use of Polymorphprep technique provides a practical and effective method for the isolation of neutrophils from peripheral blood which can be used for investigative purposes. Validation of the purity of the isolated cell population can be readily confirmed using standard clinical laboratory cell counting and differential analysis.

The serum glycoprotein fetuin-A promotes Lewis lung carcinoma tumorigenesis via adhesive-dependent and adhesive-independent mechanisms.

Fetuin-A is a serum glycoprotein in the cystatin family associated with the regulation of soft tissue calcification. We tested the role of systemic fetuin in tumor cell growth and metastasis by injecting Lewis lung carcinoma (LLC) cells into fetuin-A null and their wild-type (WT) littermate control C57BL/6 mice via the tail vein, s.c., and intrasplenic routes. In the experimental metastasis assay, the lungs of the WT mice were filled with metastatic nodules, whereas the lungs of the fetuin-A null mutant mice were virtually free of colonies at the end of 2 weeks. Lung colonization responded to the levels of serum fetuin-A in a dose-dependent manner, as observed by the formation of half as many colonies in mice heterozygous for the fetuin-A locus compared with homozygous WT mice and restoration of lung colonization by the administration of purified fetuin-A to fetuin-A-null mice. Serum fetuin-A also influenced the growth of LLC cells injected s.c.: fetuin-A-null mice developed small s.c. tumors only after a substantial delay. Similarly, intrasplenic injection of LLC cells resulted in rapid colonization of the liver with metastasis to the lungs within 2 weeks in the WT but not fetuin-A null mice. To examine the mechanism by which fetuin-A influences LLC colonization and growth, we showed that LLC tumor cells adhere to fetuin-A in a Ca(2+)-dependent fashion, resulting in growth of the tumor cells. These studies support the role of fetuin-A as a major growth promoter in serum that can influence tumor establishment and growth.