Galectin-1 is overexpressed in nasal polyps under budesonide and inhibits eosinophil migration

Because of the importance of galectins for various cellular activities, the influence of the glucocorticoid budesonide on the level of expression of galectins-1 and -3 was investigated in human nasal polyposis. Ten nasal polyps obtained from surgical resection were maintained for 24 hours in the presence of various concentrations of budesonide. As quantitatively demonstrated by means of computer-assisted microscopy, 250 ng/ml (the highest dose tested) induced a pronounced increase of galectin-1 expression. This feature was observed in nasal polyps from allergic patients but not in those from nonallergic patients. Since eosinophils represent the main inflammatory cell population in nasal polyps, we investigated the effect of galectin-1 on their migration levels by means of quantitative phase-contrast computer-assisted videomicroscopy. Our results show that galectin-1 (coated on plastic supports) markedly reduced the migration levels of eosinophils in comparison to P-selectin. On the cellular level, marked modifications in the polymerization/depolymerization dynamics of the actin cytoskeleton (as revealed by means of computer-assisted fluorescence microscopy) and, to a much lesser extent, an increase in the adhesiveness of eosinophils to tested substrata were detectable. The present study therefore reveals a new galectin-1-mediated mechanism of action for glucocorticoid-mediated anti-inflammatory effects.     

Pathogenic role of P-selectin in experimental cerebral malaria: importance of the endothelial compartment

P-selectin is a leukocyte adhesion receptor expressed on the surface of activated platelets and endothelial cells. Its role in the pathogenesis of cerebral malaria was explored in a murine model of cerebral malaria. Infection of mice with Plasmodium berghei ANKA led to P-selectin up-regulation in brain vessels of cerebral malaria-susceptible mice but not of cerebral malaria-resistant mice. Treatment of susceptible mice with anti-mouse P-selectin mAb failed to prevent the development of the neurological syndrome. However, P-selectin-deficient mice infected with Plasmodium berghei ANKA had a cumulative incidence of cerebral malaria which was significantly reduced compared to wild-type animals (4.5% versus 80%, respectively), despite identical levels of parasitemia, platelet and leukocyte accumulation. To determine whether P-selectin on platelets and/or endothelium was responsible for the microvascular pathology, cerebral malaria was assessed in chimeric mice deficient in platelet or endothelial P-selectin, which were generated by bone marrow transplantation. Mice deficient only in endothelial P-selectin did not show any sign of cerebral malaria (vascular plugging, hemorrhages, or edema), while mice lacking only platelet P-selectin showed signs of cerebral malaria similar to that seen in wild-type mice. These results indicate that endothelial P-selectin plays an important role in the pathogenesis of cerebral malaria.     

Anatomical Mapping of the External Obturator Footprint: A Study In Cadavers with Implications for Direct Anterior THA

BackgroundThe external obturator footprint in the trochanteric fossa has been suggested as a potential landmark for stem depth in direct anterior THA. Its upper border can be visualized during surgical exposure of the femur. A recent study reported that the height of the tendon has little variability (6.4 ± 1.4 mm) as measured on CT scans and that the trochanteric fossa is consistently visible on conventional pelvic radiographs. However, it is unclear where exactly the footprint of this tendon should be templated during preoperative planning so that it can be useful intraoperatively.Questions/purposesIn this study, we sought: (1) to provide instructions on exactly where to template the external obturator footprint on a preoperative planning radiograph, and (2) to confirm the small variability in height of the external obturator footprint found on CT scans in a cadaver study.MethodsTwo-dimensional (2-D) and three-dimensional (3-D) imaging was used to map the anatomy of the external obturator footprint. This dual approach was chosen because of their complementarity; conventional 2-D radiographs translate to clinical practice but 3-D navigation-based digitalization combined with CT allows for a better understanding of the cortical lines that comprise the outline of the trochanteric fossa. In 12 (four males, mean age 80 years, range 69 to 88) formalin-treated cadaveric lower extremities including the pelvis, the external obturator tendon was dissected, and the top and bottom end of its footprint marked with two small needles, and calibrated radiographs were taken. For another five (three males, mean age 75.7 years, range 61 to 91) fresh-frozen cadaveric lower extremities, including femoral reflective marker frames, CT scans were obtained and the exact location of the external obturator footprint was recorded using 3-D navigation-based digitalization. Qualitative analysis of both imaging modalities was used to develop instructions on where the external obturator footprint should be templated on a preoperative planning radiograph. Quantitative analysis of the dimensions of the external obturator footprint was performed.ResultsThe lowest point of the external obturator footprint was consistently found (± 1 mm) at the intersection of the vertical line comprised of the lateral wall of the trochanteric fossa and the oblique line formed by the intertrochanteric crest and therefore allows templating of this structure on the preoperative planning radiograph. The median (range) height of the footprint measured 6.4 mm and demonstrated small variability (4.7 to 7.6).ConclusionsWe suggest templating a 6.4-mm circle with its bottom on the intersection described above.Clinical RelevanceThe distance between the templated shoulder of the stem and the top of the circle can be used intraoperatively for guidance. Discrepancy should lead to re-evaluation of stem depth and leg length. Future work will investigate the usability, validity, and reliability of the proposed methodology in daily clinical practice.     

Hospital-Acquired Pneumonia in Critically Ill Patients

Study objectives: To identify, in patients experiencing hospital-acquired pneumonia (HAP), prognostic factors present at disease onset and build an algorithm capable of stratifying mortality risk upon HAP onset. Design: Observational cohort from January 1994 to December 2001. Setting: One intensive care unit (ICU) from a university-affiliated, urban teaching hospital. Patients: All consecutive patients exhibiting bacteriologically documented HAP either on ICU admission or during ICU stay. Interventions: Data collection and multivariate analysis using Chi-Square Automatic Interaction and Detection technique. Results: 168 patients were studied. The overall mean mortality rate was 49.4%. Upon onset of HAP, five independent variables allowed binary stratification of mortality risk. These consisted of underlying diseases (nonfatal versus ultimately and rapidly fatal diseases), Simplified Acute Physiology Score II (less than versus ≥37), platelet count (less than versus ≥150 000/mm³), chest x-ray involvement (1 versus >1 lobe), and PaO₂/FiO₂ (less than versus ≥167mm Hg). A branching algorithm consisting of these five variables identified patients with HAP at both low (<35%) and high (>75%) risk of mortality. Conclusion: Mortality in ICU patients with HAP may be predicted early, upon onset of HAP, by the cumulative use of prognostic factors in an algorithm.     

Pulmonary sarcoidosis with an alveolar radiographic pattern.

Thirty-three cases of sarcoidosis (4.4% of 746 patients) showed an alveolar radiological pattern. A study of pulmonary function was carried out in 25 patients and compared with that of 46 patients with the interstitial radiological type of sarcoidosis. Twenty-two cases have been followed up from one to six years after the initial examination. The radiographic lesions were most often bilateral and included nodules greater than 15 mm with ill-defined margins or diffuse, infiltrative, non-retractile opacities with fluffy margins. Bilateral mediastinal lymph nodes were present in 27 patients. In 20 patients an associated reticulation was found on radiography. In four patients an open lung biopsy was done. The granulomatous nodules were identical to those found in other forms of sarcoidosis, although they were more confluent in the affected areas. Clinical and functional findings did not differ from those in the more common forms of sarcoidosis. Alveolar sarcoidosis has a sudden course. The alveolar radiological patterns always disappeared, with or without steroid treatment, while reticular patterns persisted in four patients. Rapid radiological changes were observed. Some functional abnormalities persisted in cases that were followed. It is concluded that alveolar sarcoidosis is a distinct acute form of sarcoidosis.     

A new Rickettsia species found in fleas collected from human dwellings and from domestic cats and dogs in Senegal

The insects of the order Siphonaptera, commonly named fleas, are vectors of pathogens around the world. Our previous studies showed that 4.4% of acute febrile diseases in the Sine-Saloum region of Senegal were due to Rickettsia felis. The aim of this study was to explain the high prevalence of R. felis infections in two rural Senegalese populations by an entomological, systematic monitoring protocol. A total of 232 fleas from three species (Ctenocephalides felis, Echidnophaga gallinacea, and Synosternus pallidus) were collected by candle trapping and manually from pets in the villages of Dielmo and Ndiop during the year 2010. The fleas were then tested for the presence of Bartonella and Rickettsia species. No fleas were found to be positive for any Bartonella species or R. felis. Surprisingly, we found that 91.4% of S. pallidus were infected by a new Rickettsia species, which, based on sequence analysis of gltA, ompB, and two fragments of rpoB, was found to be closely related to R. felis. The results from this study did not explain the high incidence of R. felis infections in these Senegalese populations.     

Activation of latent transforming growth factor beta 1 and inhibition of matrix metalloprotease activity by a thrombospondin-like tripeptide linked to elaidic acid

Impaired wound healing and skin aging are characterized by neutral protease-mediated destruction of matrix macromolecules associated with disturbance in tissue repair. We synthesized a fatty acyl-peptide derivative at aims to simultaneously activate latent TGF-beta through its peptide domain, KFK, and inhibit MMPs through its lipophilic moiety, elaidic acid. Elaidyl-KFK as well as KFK were shown to activate LAP-TGF-beta both in vitro, using a solid phase assay with immobilized LAP-TGF-beta, and ex vivo using human dermal fibroblasts cultures. In both assays, as much as up to 10% of LAP-TGF-beta added could be recovered as active form. KQK, KQFK as well as their lipopeptide counterparts were inactive. Elaidyl-KFK-mediated LAP-TGF-beta activation led to up-regulation of collagen and TIMP-1 production and down regulation of PMA-induced MMP-1 expression in fibroblasts cultures. Those effects could be suppressed by supplementing cell culture medium with blocking TGF-beta antibody. Elaidyl-KFK inhibited MMP-2, MMP-9, MMP-3, MMP-1, in vitro with IC(50) equal to 1.2, 1.0, 0.24 and 8.9 microM, respectively. Its ex vivo inhibitory capacity, as assessed using skin tissue sections, towards the elastin-degrading capacity of MMP-9 was even more pronounced. At a 1 microM concentration, the lipopeptide decreased by up to 80% enzyme activity. Thus, "Lipospondin," i.e. elaidyl-KFK might be considered as a promising model compound to prevent age-associated dermal alterations.     

A survey of newborn screening for cystic fibrosis in Europe.

BACKGROUND: Cystic fibrosis (CF) is a recessively inherited condition caused by mutation of the CFTR gene. Newborn infants with CF have raised levels of immuno-reactive trypsinogen (IRT) in their serum. Measurement of IRT in the first week of life has enabled CF to be incorporated into existing newborn screening (NBS) blood spot protocols. However, IRT is not a specific test for CF and NBS therefore requires a further tier of tests to avoid unnecessary referral for diagnostic testing. Following identification of the CFTR gene, DNA analysis for common CF-associated mutations has been increasingly used as a second tier test. The aim of this study was to survey the current practice of CF NBS programmes in Europe. METHOD: A questionnaire was sent to 26 regional and national CF NBS programmes in Europe. RESULTS: All programmes responded. The programmes varied in number of infants screened and in the protocols employed, ranging from sweat testing all infants with a raised first IRT to protocols with up to four tiers of testing. Three different assays for IRT were used; in the majority (24) this was a commercially available kit (Delfia). A number of programmes employed a second IRT measurement in the 4th week of life (as the IRT is more specific at this point). Nineteen programmes used DNA analysis for common CFTR mutations on samples with a raised first IRT. Three programmes used a second IRT measurement on infants with just one recognised mutation to reduce the number of infants referred for sweat testing. Referral to clinical services was prompt and diagnosis was confirmed by sweat testing, even in infants with two recognised mutations in most programmes. Subsequent clinical pathways were less uniform. Multivariate analysis demonstrated a relationship between the age of diagnosis and the timing of the first IRT. More sweat tests were undertaken if the first IRT was earlier and the diagnosis was later. CONCLUSIONS: Annually these programmes screen approximately 1,600,000 newborns for CF and over 400 affected infants are recognised. The findings of this survey will guide the development of European evidence based guidelines and may help new regions or nations in the development and implementation of NBS for cystic fibrosis.