Mechanisms of non-opsonized zymosan-induced and luminol-enhanced chemiluminescence in whole blood and isolated phagocytes

A luminol-dependent non-opsonized zymosan-induced chemiluminescence method for phagocytes in small quantities of whole blood (40 microliters; final dilution: 1:14) is described. It was characterized with reference to cellular and humoral components, and also applied to isolated neutrophils, eosinophils and monocytes. Normal values for whole blood chemiluminescence and for neutrophils, eosinophils and monocytes are presented. From the chemiluminescence characteristic of distinct phagocytes and their frequency distribution pattern in whole blood, it is concluded that whole blood chemiluminescence has its source predominantly in neutrophils. The question as to the origin of chemiluminescence in phagocytes of whole blood and isolated neutrophils is investigated. The results support the importance of the myeloperoxidase-H2O2-halide system, but also go beyond this. The release of arachidonic acid by phospholipase A2 and of diacylglycerol and inositol trisphosphate by phospholipase C, the metabolism of arachidonic acid by the cyclooxygenase and lipoxygenase pathway, the activation of membrane NADPH oxidase by diacylglycerol and the calcium mobilisation by inositol trisphosphate are necessary for the chemiluminescence reaction. Inhibition of either mechanism suppresses the chemiluminescence response. The interaction of non-opsonized zymosan with plasma opsonins, phagocyte Fc- and complement receptors, respectively, for the initiation of chemiluminescence, was investigated. Non-opsonized zymosan initiates a chemiluminescence response in blood phagocytes in the absence of opsonin from the interaction of the zymosan polysaccharide component glucan with the complement receptor type 3. In the presence of plasma this receptor type also mediates the major chemiluminescence response brought about by the zymosan-coated cleavage products of complement fraction three, iC3b and to a minor degree C3b, while immunoglobulin G-coated zymosan interaction with the Fc-receptor is in this case of minor importance.     

3D‐electron microscopic characterization of interstitial cells in the human bladder upper lamina propria

Aims

To explore the ultrastructure of interstitial cells in the upper lamina propria of the human bladder, to describe the spatial relationships and to investigate cell‐cell contacts.

Methods

Focused ion beam scanning electron microscopy (FIB‐SEM), 3‐View SEM and confocal laser scanning microscopy were used to analyze the 3D ultrastructure of the upper lamina propria in male and female human bladders.

Results

3View‐SEM image stacks as large as 59 × 59 × 17 μm³ (xyz) at a resolution of 16 × 16 × 50 nm³ and high resolution (5 × 5 × 10 nm³) FIB‐SEM stacks could be analyzed. Interstitial cells with myoid differentiation (mIC) and fibroblast like interstitial cells (fIC) were the major cell types in the upper lamina propria. The flat, sheet‐like ICs were oriented strictly parallel to the urothelium. No spindle shaped cells were present. We furthermore identified one branched cell (bIC) with several processes contacting urothelial cells by penetrating the basal membrane. This cell did not make any contacts to other ICs within the upper lamina propria. We found no evidence for the occurrence of telocytes in the upper lamina propria.

Conclusions

Comprehensive 3D‐ultrastructural analysis of the human bladder confirmed distinct subtypes of interstitial cells. We provide evidence for a foremost unknown direct connection between a branched interstitial cell and urothelial cells of which the functional role has still to be elucidated. 3D‐ultrastructure analyses at high resolution are needed to further define the subpopulations of lamina propria cells and cell‐cell interactions.     

Isolation of Stenotrophomonas maltophilia in asymptomatic lung transplant recipients: effects of treatment on eradication and outcome

In this retrospective, single‐center data analysis, we audited our clinical practice to treat Stenotrophomonas maltophilia in asymptomatic lung transplant recipients (LTRs). Eighteen LTRs with confirmed isolation of S. maltophilia were identified. Twelve of these LTRs have been treated with antibiotics, while 6 were managed without treatment. Treatment was based on antibiograms (trimethoprim/sulfamethoxazole [TMP/SMX] (8/12), levofloxacin (1/12), or both (3/12). Clearance (12/12 vs 6/6), eradication (10/12 vs 3/6, P=.27), and freedom from S. maltophilia recurrence (83%±11% vs 40%±22% after one year, log‐rank P=.09) were not found to differ significantly between treated and untreated patients. None of the patient groups showed significant changes in lung function or biochemical variables. Creatinine levels at the end of the study period were found to be higher in treated patients compared to the untreated group (P=.049). De novo acquired TMP/SMX resistance in S. maltophilia strains was not observed. These results indicate no evidence that antibiotic treatment for S. maltophilia in asymptomatic LTRs alters lung function or the clinical outcome.     

Solid-State Quantitative <Superscript>1</Superscript>H and <Superscript>31</Superscript>P MRI of Cortical Bone in Humans

Magnetic resonance imaging (MRI) plays a pivotal role for assessment of the musculoskeletal system. It is currently the clinical modality of choice for evaluation of soft tissues including cartilage, ligaments, tendons, muscle, and bone marrow. By comparison, the study of calcified tissue by MRI is still in its infancy. In this article, we review the potential of the modality for assessment of cortical bone properties known to be affected in degenerative bone disease, with focus on parameters related to matrix and mineral densities, and porosity, by means of emerging solid-state ¹H and ³¹P MRI techniques. In contrast to soft tissues, the MRI signal in calcified tissues has very short lifetime, on the order of 100 μs to a few milliseconds, demanding customized imaging approaches that allow capture of the signal almost immediately after excitation. The technologies described are suited for quantitatively imaging human cortical bone in specimens as well as in vivo in patients on standard clinical imagers, yielding either concentrations in absolute units when measured against a reference standard, or more simply, in the form of surrogate biomarkers. The two major water fractions in cortical bone are those of collagen-bound and pore water occurring at an approximately 3:1 ratio. Collagen-bound water density provides a direct quantitative measure of osteoid density. While at an earlier stage of development, quantification of mineral phosphorus by ³¹P MRI yields mineral density and, together with knowledge of matrix density, should allow quantification of the degree of bone mineralization.     

Prolonged Mechanical Ventilation After Lung Transplantation—A Single‐Center Study

This single‐center study examines the incidence, etiology, and outcomes associated with prolonged mechanical ventilation (PMV), defined as time to definite spontaneous ventilation >21 days after double lung transplantation (LTx). A total of 690 LTx recipients between January 2005 and December 2012 were analyzed. PMV was necessary in 95 (13.8%) patients with decreasing incidence during the observation period (p < 0.001). Independent predictors of PMV were renal replacement therapy (odds ratio [OR] 11.13 [95% CI, 5.82–21.29], p < 0.001), anastomotic dehiscence (OR 8.74 [95% CI 2.42–31.58], p = 0.001), autoimmune comorbidity (OR 5.52 [95% CI 1.86–16.41], p = 0.002), and postoperative neurologic complications (OR 5.03 [95% CI 1.98–12.81], p = 0.001), among others. Overall 1‐year survival was 86.0% (90.4% for LTx between 2010 and 2012); it was 60.7% after PMV and 90.0% in controls (p < 0.001). Conditional long‐term outcome among hospital survivors, however, did not differ between the groups (p = 0.78). Multivariate analysis identified renal replacement therapy (hazard ratio [HR] 3.55 [95% CI 2.40–5.25], p < 0.001), post‐LTx extracorporeal membrane oxygenation (HR 3.47 [95% CI 2.06–5.83], p < 0.001), and prolonged inotropic support (HR 1.95 [95% CI 1.39–2.75], p < 0.001), among others, as independent predictors of mortality. In conclusion, PMV complicated 14% of LTx procedures and, although associated with increased in‐hospital mortality, outcomes among patients surviving to hospital discharge were unaffected.     

Quality assurance in stereotactic radiosurgery/radiotherapy according to DIN 6875-1

The new DIN ('Deutsche Industrie-Norm') 6875-1, which is currently being finalised, deals with quality assurance (QA) criteria and tests methods for linear accelerator and Gamma Knife stereotactic radiosurgery/radiotherapy including treatment planning, stereotactic frame and stereotactic imaging and a system test to check the whole chain of uncertainties. Our existing QA program, based on dedicated phantoms and test procedures, has been refined to fulfill the demands of this new DIN. The radiological and mechanical isocentre corresponded within 0.2 mm and the measured 50% isodose lines were in agreement with the calculated ones within less than 0.5 mm. The measured absorbed dose was within 3%. The resultant output factors measured for the 14-, 8- and 4-mm collimator helmet were 0.9870 +/- 0.0086, 0.9578 +/- 0.0057 and 0.8741 +/- 0.0202, respectively. For 170 consecutive tests, the mean geometrical accuracy was 0.48 +/- 0.23 mm. Besides QA phantoms and analysis software developed in-house, the use of commercially available tools facilitated the QA according to the DIN 6875-1 with which our results complied.     

Lymph node surgery in papillary thyroid carcinoma

BACKGROUND: The impact of nodal disease remains controversial in papillary thyroid carcinoma (PTC). STUDY DESIGN: One surgeon treated 159 unselected patients, who were followed up for 1 to 27 years. We present a retrospective analysis with respect to nodal disease. Occult nodal disease was investigated, including metachronous nodal disease (mpN(1)) in primarily node negative patients (pN(0), clinical [c]N(0)). RESULTS: Therapeutic lymphadenectomies, prophylactic lymphadenectomies, or no lymphadenectomy were carried out in 42 (cN(1)), 29 (cN(0)), and 88 (cN(0)) patients, respectively, with stage pN(1) in 41 (98%), in 5 (17%), and in 2 (2.3%) patients, respectively (17% versus 2.3% p < 0.005). Sensitivity and specificity of clinical staging were 85% and 99%, respectively. More frequent prophylactic lymphadenectomy during the study period (p = 0.002) led to a nonsignificant increase in stage pN(1) (26% versus 30%). Immunohistochemistry led to upstaging of only 3% of histologically negative nodes and one (4%) pN(0) patient. Nodal recurrence occurred in 8 of 156 patients (5%) treated for cure, in 12% of pN(1) versus 3% of pN(0) cN(0) tumors (p = 0.009), in 15% of TNM high-versus 3% of low-risk patients (p = 0.006), and in 5% each of patients, younger than 45 and 45 years or more. In TNM high-risk patients, tumor-related survival was 50% for stage pN(1) versus 86% for stage pN(0), cN(0) (p = 0.03) (100% and 100% in low-risk patients). CONCLUSIONS: The rate of occult nodal disease might be relatively low, and it does not frequently progress to clinical recurrent disease. Clinical nodal status might be valid for deciding the extent and radicality of node dissection. Prophylactic (central) lymphadenectomy should be carried out without radicality-associated morbidity. Macroscopic nodal disease warrants more rigorous, compartment-oriented lymphadenectomy. There is no rationale for detection of occult disease and micrometastasis by frozen section or immunohistochemistry.     

Prospective study of early rheumatoid arthritis. I. Prognostic value of IgA rheumatoid factor.

Thirty-three patients with early arthritis, 28 of whom developed classical/definite rheumatoid arthritis (RA), were followed up for two to four years. Rheumatoid factor (RF) levels of the IgM, IgA, and IgG isotypes were measured in serum and synovial fluid by an ELISA technique developed in our laboratory. All seven patients who presented with raised IgA RF developed erosions of their hands and wrists. This was significantly different from the remaining 26. By contrast none of the five patients who presented with isolated elevation of IgM RF developed erosive disease. The patients with raised IgA RF needed significantly more treatment with 'specific' drugs than the remaining 26. It is suggested that the detection of IgA RF in early RA indicates poor prognosis, justifying a more aggressive treatment at an early stage.     

Metastatic tumors to the parotid and submandibular glands--analysis and differential diagnosis of 108 cases

The distinction between primary salivary gland tumors and metastases of other primary tumors in salivary glands is of special importance for therapy and prognosis. In the files of the Salivary Gland Register, 10,944 cases were collected during 1965 and 1985. Among these cases, there were 108 cases of metastatic tumors to the parotid and submandibular gland. The pathohistological analysis of these tumors revealed the following data: 47 cases (43%) of metastatic tumors were localized in the parenchyma of the parotid gland (37 cases) or of the submandibular gland (10 cases). 61 cases (57%) displayed metastases in the lymph nodes of the parotid gland (38 cases) or of the submandibular gland (23 cases). The sublingual gland was free of metastatic tumors. 65 metastatic tumors originated from primary tumors in the neighborhood (head and neck). 32 tumors were carcinomas of the skin, 17 tumors were melanomas, and 13 tumors were nasopharyngeal cancers. Metastases of thyroid cancers were found in 3 cases. The relative frequency of metastases in the lymph nodes of the salivary glands is due to the intense drainage with lymph vessels and the presence of many lymph nodes which are localized especially in the gland parenchyma or around the parotid gland. 21 metastatic tumors originated from primary tumors distant from the head and neck region. There were metastases of lung cancers (7 cases), renal cancers (6 cases), mammary cancers (6 cases), colonic cancer (1 case) and uterus cancer (1 case). Clear cell carcinomas in salivary gland tissue should always be checked for a metastasis of a primary renal cancer.(ABSTRACT TRUNCATED AT 250 WORDS)