CD133-enriched CD34(-) (CD33/CD38/CD71)(-) cord blood cells acquire CD34 prior to cell division and hematopoietic activity is exclusively associated with CD34 expression

We compared the cell division behavior of CD34(-) and CD34(+) (CD33/CD38/CD71)-negative (Lin(-)) CD133(+) cord blood cells stimulated with the cytokines Flt3-ligand, stem cell factor, and thrombopoietin. Within a 4-day time frame, Lin(-)CD34(-) CD133(+) (CD34(-)) cells underwent more cell divisions in serum-free culture than their Lin(-)CD34(+) CD133(+) (CD34(+)) counterparts. The majority of CD34(-) cells acquired expression of CD34 in vitro, including most undivided cells. Moreover, hematopoietic activity from both CD34(-) and CD34(+) cells was exclusively retained within the cell fraction expressing CD34 after 4 days in culture. Most strikingly, in cultures from Lin(-)CD34(-) cells hematopoietic activity was associated with the fraction of divided cells, whereas in cultures of CD34(+) cells, hematopoietic activity associated with the undivided cell fraction. Therefore, clonogenic CD34(+) cells either do not divide or lose their clonogenic capacity upon cell division in vitro, while CD34(-) cells divide and retain this capacity under the same specific conditions. In conclusion, we demonstrate that CD133-enriched Lin(-)CD34(-) cord blood cells acquire CD34 prior to cell division and that long-term hematopoietic activity is associated exclusively with expression of CD34.     

Absence of mutations in the transforming growth factor-beta type II receptor in sporadic lung cancers with microsatellite instability and rare H-ras1 alleles

The transforming growth factor-beta type II receptor (RII) is commonly mutated in colon and gastric cancers with microsatellite instability (MI). We utilized our series of lung cancers with MI and rare alleles of the H-ras1 gene to determine the association between MI and RII mutations and searched the entire RII coding region in 33 lung cancers with MI by polymerase chain reaction-single-strand conformation polymorphism analysis. We found no mutations, and these data support other recent evidence that RII mutations rarely occur except in colon and gastric tumors with MI.      

Segmentation of the Visible Human for high-quality volume-based visualization

This article describes a combination of interactive classification and super-sampling visualization algorithms that greatly enhances the realism of 3-D reconstructions of the Visible Human data sets. Objects are classified on the basis of ellipsoidal regions in RGB space. The ellipsoids are used for super-sampling in the visualization process.     

Selective expression of IL-7 receptor on memory T cells identifies early CD40L-dependent generation of distinct CD8+ memory T cell subsets

Several recent studies have demonstrated that T-helper cell-dependent events during the initial priming period are required for the generation of CD8(+) T cell-mediated protective immunity. The underlying mechanisms of this phenomenon have not yet been determined, mostly because of difficulties in studying memory T cells or their precursor populations at early stages during immune responses. We identified IL-7 receptor (CD127) surface expression as a marker for long-living memory T cells, most importantly allowing the distinction between memory and effector T cells early after in vivo priming. The combination of surface staining for CD127 and CD62L further separates between two functionally distinct memory cell subsets, which are similar (if not identical) to cell subsets recently described as central memory T cells (CD127(high) and CD62L(high)) and peripheral effector memory T cells (CD127(high) and CD62L(low)). Using this new tool of memory T cell analysis, we demonstrate that CD8(+) T cell priming in the absence of T cell help or CD40L specifically alters the generation of the effector memory T cell subset, which appears to be crucial for immediate memory responses and long-term maintenance of effective protective immunity. Our data reveal a unique strategy to obtain information about the quality of long-term protective immunity early during an immune response, a finding that may be applied in a variety of clinical settings, including the rapid monitoring of vaccination success.     

The Antitumorigenic Function of EGFR in Metastatic Breast Cancer is Regulated by Expression of Mig6

Numerous studies by our lab and others demonstrate that epidermal growth factor receptor (EGFR) plays critical roles in primary breast cancer (BC) initiation, growth and dissemination. However, clinical trials targeting EGFR function in BC have lead to disappointing results. In the current study we sought to identify the mechanisms responsible for this disparity by investigating the function of EGFR across the continuum of the metastatic cascade. We previously established that overexpression of EGFR is sufficient for formation of in situ primary tumors by otherwise nontransformed murine mammary gland cells. Induction of epithelial-mesenchymal transition (EMT) is sufficient to drive the metastasis of these EGFR-transformed tumors. Examining growth factor receptor expression across this and other models revealed a potent downregulation of EGFR through metastatic progression. Consistent with diminution of EGFR following EMT and metastasis EGF stimulation changes from a proliferative to an apoptotic response in in situ versus metastatic tumor cells, respectively. Furthermore, overexpression of EGFR in metastatic MDA-MB-231 BC cells promoted their antitumorigenic response to EGF in three dimensional (3D) metastatic outgrowth assays. In line with the paradoxical function of EGFR through EMT and metastasis we demonstrate that the EGFR inhibitory molecule, Mitogen Induced Gene-6 (Mig6), is tumor suppressive in in situ tumor cells. However, Mig6 expression is absolutely required for prevention of apoptosis and ultimate metastasis of MDA-MB-231 cells. Further understanding of the paradoxical function of EGFR between primary and metastatic tumors will be essential for application of its targeted molecular therapies in BC.Abbreviations: Mig6, mitogen-induced gene-6; BC, breast cancer; EGFR, epidermal growth factor receptor; Mig6-EBR, EGFR binding region of Mig6; TGF-β, transforming growth factor-β; WT, wild type; NMuMG, normal murine mammary gland cells; NME, NMuMG cells transformed by EGFR overexpression; CCLE, Cell Line Encyclopedia; MET, mesenchymal-epithelial transition     

Impact of patient-prosthesis mismatch and aortic valve design on coronary flow reserve after aortic valve replacement.

OBJECTIVES: This prospective-randomized study investigated the effect of aortic valve design and patient-prosthesis mismatch (PPM) on coronary flow reserve (CFR) after mechanical or biological aortic valve replacement (AVR) in patients with aortic stenosis (AS). BACKGROUND: Coronary flow reserve may be an important parameter of long-term survival after AVR in patients with AS. Reduced CFR may contribute to more cardiovascular events and greater rates of mortality. METHODS: A total of 48 patients undergoing AVR underwent magnetic resonance imaging for the measurement of coronary flow preoperatively, 5 days postoperatively, and at 6-month follow-up with measurement of CFR. Patients scheduled for mechanical AVR were randomized to a tilting disc or bileaflet prosthesis (n = 12 in each group). For biological AVR, patients were scheduled to receive a stented (n = 12) or stentless (n = 12) valve. Patients also underwent echocardiography with measurement of transvalvular pressure gradients and left ventricular mass regression. RESULTS: Postoperatively, coronary flow increased significantly in all groups (p < 0.001). Only stentless valves demonstrated a normal CFR (3.4 +/- 0.3 vs. 2.3 +/- 0.1 for stented biological valves, 2.1 +/- 0.2 for tilting disc, and 2.2 +/- 0.3 for bileaflet mechanical valves). Patient-prosthesis mismatch with an indexed effective orifice area <0.85 cm2/m2 led to decreased rates of CFR in the tilting disc, stentless, and stented groups. Pressure gradients were 14 +/- 3 mm Hg for tilting disc, 12 +/- 4 mm Hg for bileaflet, 19 +/- 6 mm Hg for stented, and 10 +/- 4 mm Hg for stentless valves. CONCLUSIONS: Normalization of CFR after AVR in patients with AS was observed only for stentless valves. Coronary flow reserve might explain the excellent long-term results for stentless valves. (Impact of Patient-Prosthesis Mismatch on Coronary Flow Reserve; http://www.clinicaltrials.gov/ct/show/NCT00310947?order=1; NCT00310947).     

Chronic diarrhea after a journey to Sri Lanka

A 51-year-old woman was referred for evaluation of chronic diarrhea. She had spent a 14 day vacation in Sri Lanka three years ago. The clinical examination of the patient was unremarkable. Values for protein, iron, zinc, copper and folic acid were decreased and the Shilling- and D-xylose tests revealed pathological results. Gliadin and Endomysium antibodies were not detectable. Histologic examination of the duodenum displayed chronic duodenitis with increased epithelial regeneration and villous atrophy. In the MRI a segment of the mid small bowel with increased thickness of the intestinal wall was described. Abdominal CT-scans demonstrated multiple, enlarged mesenteric lymph nodes. Laparoscopy with biopsies of the ileum and mesenteric lymph nodes excluded a malignant lymphoma, mycobacteriosis or Whipple's disease. Oral therapy with tetracyclines (250 mg q. i. d.) and substitution of folic acid and iron led to rapid improvement of the clinical symptoms which persisted after cessation of the antibiotic therapy. In view of the clinical course tropical sprue has to be assumed despite the short duration of the journey to a tropical region.