Role and Localization of Urokinase Receptor in the Formation of New Microvascular Structures in Fibrin Matrices

Fibrin or a fibrinous exudate can facilitate angiogenesis in many pathological conditions. In vitro, the outgrowth of capillary-like structures in fibrin can be mimicked by exposing human microvascular endothelial cells (hMVECs) to an angiogenic growth factor and tumor necrosis factor (TNF)-alpha. Urokinase-type plasminogen activator (u-PA) and plasmin activities are required for this angiogenic process. This study focuses on the role and localization of the u-PA receptor (u-PAR) in newly formed microvascular structures. The u-PAR-blocking monoclonal antibody (MAb) H-2 completely inhibited the formation of capillary-like tubular structures induced by exposure of hMVECs to basic fibroblast growth factor and TNF-alpha. This was accompanied by a several-fold increase in u-PA accumulation in the conditioned medium. The effect of MAb H-2 was not caused by blocking cellular activation by u-PA/u-PAR interaction, as the amino-terminal fragment (ATF) of u-PA, which also activates u-PAR, prevented tube formation. In addition, the inhibition by MAb H-2 was not due to an effect of the antibody on u-PAR-vitronectin binding. These data show that inhibition of tube formation can be caused not only by inhibition of u-PA or plasmin activities but also by unavailability of the u-PAR for cell-bound proteolysis. Immunohistochemical analysis showed that in in vitro angiogenesis u-PAR and u-PA were localized on the invading, tube-forming hMVECs and not on the endothelial cells that are located on top of the fibrin matrix. u-PAR and u-PA were also prominently expressed on endothelial cells of neovessels present in an atherosclerotic plaque. These data may give more insight into the role of u-PAR in repair-associated angiogenesis.     

Characterization of trisomy 8 in pediatric undifferentiated sarcomas using advanced molecular cytogenetic techniques

Pediatric undifferentiated soft tissue sarcomas (USTS) are a rare group of neoplasms that are unclassifiable despite the application of immunohistochemical, cytogenetic, and molecular techniques. To date, there is a dearth of studies looking at the cytogenetic and molecular genetic alterations in such tumors. Trisomy 8, a frequent molecular alteration in neoplasia, is seen in several soft tissue sarcomas, including Ewing sarcoma/primitive neuroectodermal tumor (ES/PNET), synovial sarcoma, and leiomyosarcoma. Because USTS share several clinicobiological features with the aforementioned tumors, the occurrence of alterations in chromosome 8 was studied in 11 pediatric USTS using a combination of interphase fluorescence in situ hybridization (FISH), spectral karyotyping (SKY), and genomic profiling with oligonucleotide array comparative genomic hybridization (aCGH). The copy number status of MYC was also assessed on the same tumors using dual-color FISH, with the aim of delineating the degree and intratumoral distribution of MYC amplification in this tumor. A near-uniform presence of an increase in MYC copy number was observed, along with an increase in chromosome 8 copy number in all the tumors. SKY and aCGH analysis of tumors exhibiting trisomy 8 confirmed the numerical imbalances. The occurrence of trisomy 8 in a subset of pediatric USTS confirms a shared genomic alteration with several other soft tissue sarcomas. Further studies are required to determine the clinical implications of such a finding.     

Combined activation and deactivation of visual cortex during tactile sensory processing

The involvement of occipital cortex in sensory processing is not restricted solely to the visual modality. Tactile processing has been shown to modulate higher-order visual and multisensory integration areas in sighted as well as visually deprived subjects; however, the extent of involvement of early visual cortical areas remains unclear. To investigate this issue, we employed functional magnetic resonance imaging in normally sighted, briefly blindfolded subjects with well-defined visuotopic borders as they tactually explored and rated raised-dot patterns. Tactile task performance resulted in significant activation in primary visual cortex (V1) and deactivation of extrastriate cortical regions V2, V3, V3A, and hV4 with greater deactivation in dorsal subregions and higher visual areas. These results suggest that tactile processing affects occipital cortex via two distinct pathways: a suppressive top-down pathway descending through the visual cortical hierarchy and an excitatory pathway arising from outside the visual cortical hierarchy that drives area V1 directly.     

Communication between human mast cells and CD4+ T cells through antigen‐dependent interactions

Mast cells (MCs) are immune cells residing in tissues where pathogens are first encountered. It has been indicated that MCs might also be involved in setting the outcome of T‐cell responses. However, little is known about the capacity of human MCs to express MHC class II and/or to capture and present antigens to CD4⁺ T cells. To study the T‐cell stimulatory potential of human MCs, CD34⁺ stem cell derived MCs were generated. These cells expressed HLA‐DR when stimulated with IFN‐γ, and, importantly, presented peptide and protein for activation of antigen‐specific CD4⁺ T cells. The interplay between MC and T cell led to increased HLA‐DR expression on MCs. MCs were present in close proximity to T cells in tonsil and expressed HLA‐DR and CD80, indicating their ability to present antigens to CD4⁺ T cells in T‐cell areas of human LNs. Our data show that MCs can present native antigens to human CD4⁺ T cells and that HLA‐DR expressing MCs are present in tonsil tissue, indicating that human MCs can directly activate T cells and provide a rationale to study the potential of MCs to prime and/or skew human T‐cell responses.     

Effect of dexamethasone on respiratory syncytial virus-induced lung inflammation in children: results of a randomized, placebo controlled clinical trial

Inflammatory mediators play a major role in the pathogenesis of respiratory syncytial virus (RSV) infection. The objective of this study was to evaluate the effect of i.v. dexamethasone on cytokine concentrations in tracheal aspirates (TA) of children with severe RSV disease and to correlate them with disease severity. Twenty-five cytokines were measured in TA obtained from children <2 yr old intubated for severe RSV disease, and enrolled in a double-blind study of i.v. dexamethasone (0.5 mg/kg; n = 22) vs. placebo (n = 19). Cytokine concentrations, measured at baseline and days 1 and 5 post-randomization using a multiplex assay, were compared within both treatment groups and correlated with: (i) tracheal white blood cell counts, (ii) tracheal RSV loads by culture and (iii) parameters of disease severity, including number of days of requirement for mechanical ventilation, intensive care unit (ICU), and hospitalization. At baseline interleukin (IL)-13 and IL-15 concentrations were significantly higher in the dexamethasone treatment group. On day 1 post-treatment, only MCP-1, eotaxin and IL-6 concentrations were significantly different but higher in the placebo group. On day 5: IL-13, IL-7, IL-8 and MIP-1α concentrations were higher in dexamethasone-treated patients. In both groups MIP-1β inversely correlated with the days of ventilator support; MIP-1α, MIP-1β and eotaxin inversely correlated with ICU days; and IL-6 inversely correlated with hospitalization regardless of the treatment assigned. Systemic administration of dexamethasone did not have a consistent effect on TA concentrations of pro-inflammatory cytokines. This may help explain, at least in part, the lack of clinical benefit of steroid treatment in children with severe RSV bronchiolitis.     

Effects of sleep deprivation on neural circulatory control

Effects of sleep deprivation on neural cardiovascular control may have important clinical implications. We tested the hypothesis that sleep deprivation increases heart rate, blood pressure, and sympathetic activity and potentiates their responses to stressful stimuli. We studied 8 healthy subjects (aged 40+/-5 years, 6 men and 2 women). Blood pressure, heart rate, forearm vascular resistance, and muscle sympathetic nerve activity were measured at rest and during 4 stressors (sustained handgrip, maximal forearm ischemia, mental stress, and cold pressor test). Measurements were obtained twice, once after normal sleep and once after a night of sleep deprivation. All measurements were obtained in a blinded, randomized manner. In comparison with normal sleep, sleep deprivation resulted in an increase in blood pressure (normal sleep versus sleep deprivation=82+/-8 versus 86+/-7 mm Hg, mean+/-SEM, P=0.012) and a decrease in muscle sympathetic nerve activity (normal sleep versus sleep deprivation=28+/-6 versus 22+/-6 bursts/min, P=0.017). Heart rate, forearm vascular resistance, and plasma catecholamines were not significantly changed by sleep deprivation, nor did sleep deprivation affect autonomic and hemodynamic responses to stressful stimuli. Sleep deprivation results in increased resting blood pressure, decreased muscle sympathetic nerve activity, and no change in heart rate. Thus, the pressor response to sleep deprivation is not mediated by muscle sympathetic vasoconstriction or tachycardia.     

Two-stage repair of extensive subglottic tracheal stenosis

Summary The authors describe an open technique that has been used over the past 25 years to reconstruct the subglottic tracheal region in two stages after extensive laryngotracheal stenosis. After submucosal resection of fibrous tissue and reconstruction of the subglottic and tracheal skeleton by means of two autologous osseous grafts, a large laryngotracheostomy is created during the initial stage. Some weeks later, in the second stage, the anterior wall is closed, using two cervical hinge-door flaps. Ten patients have undergone this procedure, with a minimum follow-up of 3 years. All of the patients were decannulated upon completion of the treatment without recurrence of stenosis during the follow-up period.     

Genetic alterations in head and neck cancer.

Mutations of the ras gene family appear to be an uncommon genetic alteration in SCCHN. A common region of DNA amplification on chromosome 11q13 has been identified in SCCHN. A cluster of proto-oncogenes (int-2, hst-1, bcl-1, prad-1) has been localized to the 11q13 region. Studies are needed to determine the critical genes in 11q13 whose expression drive the amplicon. Mutations of the p53 tumor suppressor gene are the most common genetic alteration in SCCHN. The hope is that dysregulated oncogenes or tumor suppressor genes may be targets for specific therapy.     

Angelchik Prosthesis Revisited

There are few long-term follow-up reports of the Angelchik prosthesis (AP). We report the longest follow-up series (66-192 months, average 145 months) to date. Between October 1983 and January 1994, 65 patients (45 men and 20 women) aged between 29 and 84 years (mean 52 years) had an AP inserted for gastro-oesophageal reflux (GOR) with or without hiatus hernia (HH). Clinical, radiological, endoscopy, and operative details were reviewed. Postoperative complications, investigations, and follow-up details were critically analyzed. All living patients (n = 53) with an AP in situ were interviewed and symptomatic assessment was carried out using a modified Visick system (I-IV). The average duration of the GOR symptoms before the operation was 5.7 years (range 10 months to 20 years). The average hospital stay was 8 days (range 5-15 days). Postoperatively, five patients developed chest infection/atelectasis, four had superficial wound infection, two had deep vein thrombosis (one with pulmonary embolism), one had urinary retention, and four developed an incisional hernia. Six patients (three with an AP in situ) died of other medical conditions. Ten (15%) patients had removal of the prosthesis. Eight (12%) and 11 (17%) had transient and persistent dysphagia, respectively. Thirteen (20%) and five (8%) patients had distal slippage and proximal migration of the prosthesis, respectively. One patient had erosion of the AP into the stomach, while in another patient, the straps of the prosthesis ruptured. Of the 53 living patients with an AP in situ, 28 (53%) were Visick I, 11 (20%) were Visick II, 11 (20%) were Visick III, and 3 (7%) were Visick IV. We conclude that the AP has poor long-term results, with only 66% attaining Visick I and II, and a prosthesis removal rate of 15% (10/65). Patients with preoperative dysphagia, hypothyroidism, and diabetes tend to do worse with an AP. Obese patients and those with failed previous fundoplication seemed to fare well with an AP. In view of poor long-term results and high incidence of complications as compared to other conventional operations for GOR, we cannot recommend the continued use of the AP.