Immunocytochemical detection of Candida albicans in formalin fixed, paraffin embedded material.
(9/2410)
AIM: To assess the ability of the commercially available monoclonal antibody 1B12 (BioGenex, San Ramon, USA) to identify C albicans in formalin fixed, paraffin wax embedded material (FFPE). METHODS: Broth cultures of 20 strains of seven Candida species were resuspended in 4% agarose blocks, fixed in formalin for 24 hours, and embedded in paraffin wax. In addition, 16 blocks of FFPE tissue known to contain periodic acid-Schiff positive fungal hyphae were examined. Antigen retrieval involved microwave treatment of specimens in citrate buffer (0.01 M; pH 6.5) before addition of 1B12 antibody for 24 hours. Bound antibody was subsequently detected using a biotinylated link antibody and a peroxidase conjugated streptavidin. RESULTS: Only C albicans strains were 1B12 positive in the agarose blocks. All FFPE tissue blocks were found to contain 1B12 positive hyphal structures, indicating the presence of C albicans. CONCLUSIONS: The ability to identify candida organisms penetrating the lesional tissue in cases of chronic hyperplastic candidosis will help to clarify the role of individual Candida spp in this important form of oral candidosis. (+info)
Proteolysis of monocyte CD14 by human leukocyte elastase inhibits lipopolysaccharide-mediated cell activation.
(10/2410)
Human leukocyte elastase (HLE), a polymorphonuclear neutrophil (PMN) serine proteinase, is proteolytically active on some membrane receptors at the surface of immune cells. The present study focused on the effect of HLE on the expression of CD14, the main bacterial lipopolysaccharide (LPS) receptor at the surface of monocytes. HLE exhibited a time- and concentration-dependent downregulatory effect on CD14 surface expression. A 30-minute incubation of 3 microM HLE was required to display 95% disappearance of the receptor. This downregulation resulted from a direct proteolytic process, not from a shedding consecutive to monocyte activation as observed upon challenge with phorbol myristate acetate (PMA). To confirm that CD14 is a substrate for HLE, this enzyme was incubated with recombinant human CD14 (Mr approximately 57,000), and proteolysis was further analyzed by immunoblot analysis. Cleavage of the CD14 molecule was directly evidenced by the generation of short-lived fragments (Mr approximately 47,000 and 30,000). As a consequence of the CD14 proteolysis, a decrease in the responsiveness of monocytes to LPS was observed, as assessed by measuring tumor necrosis factor-alpha (TNF-alpha) formation. This inhibition was only observed with 1 ng/ml of LPS, i.e., when only the CD14-dependent pathway was involved. At a higher LPS concentration, such as 10 microgram/ml, when CD14-independent pathways were operative, this inhibition was overcome. The direct proteolysis by HLE of the membrane CD14 expressed on monocytes illustrates a potential anti-inflammatory effect of HLE through inhibition of LPS-mediated cell activation. (+info)
Experience with external quality control in spermatology.
(11/2410)
Results are presented from participation in an external quality control (EQC) programme for semen analysis (UK NEQAS). Formalin-fixed semen samples and videotapes of motile spermatozoa were distributed four times a year over a 3-4 year period. Over the entire period there was close agreement for sperm concentration with, initially, the average of values from the other groups participating in the scheme, and later, values designated as reference values obtained from six laboratories of several chosen that consistently agreed with each other. The initial underestimation of the percentage of normal forms was abolished at the time of change in derivation of designated values and this largely eliminated the difference to establish closer agreement with the designated values. A consistent bias in the assessment of different categories of progressive sperm motility appeared to be resolved by a conscious decision to consider most spermatozoa as grade b and the exceptions as grade a, rather than the converse. Feedback of results to the technicians of the laboratory participating in an external quality control programme leads to reappraisal of subjective evaluation and to harmonization of results between laboratories. (+info)
A possible evolutionary role of formaldehyde.
(12/2410)
Formaldehyde is a compound which is believed to have had a role in evolutionary processes. On the other hand, the (methyl)glyoxalase pathway is a route being present in all biological organisms whereas its function has not yet been recognized in the biochemical machinery. In this article it is raised that (methyl)glyoxalase path might have functioned as a bridge between formose and archaic reductive citric acid cycles in surface metabolists at the early stage of evolution. According to the theory, formaldehyde was essential for the mentioned system as a raw molecule. Based on thermodynamic calculations a simple way of regulation is also shown. The simplicity of the theory may be in a good agreement with and an explanation of why the (methyl)glyoxalase system is of ubiquitous nature. (+info)
Polymerase chain reaction detection of Puumala virus RNA in formaldehyde-fixed biopsy material.
(13/2410)
BACKGROUND: Infections with hantaviruses, mainly Clethrionomys-derived Puumala viruses, are known causes of acute renal failure [hemorrhagic fever with renal syndrome (HFRS)] in western Europe. Laboratory diagnosis is primarily based on serology. At the time of clinical symptoms, viral RNA can hardly be detected in the blood or urine, indicating that polymerase chain reaction (PCR) is of little diagnostic value for these infections. Biopsy material is usually formaldehyde-fixed and, thus, regarded as poor quality for PCR applications. The aim of this study was to establish a technique to retrieve such material for laboratory diagnostic. METHODS: Formaldehyde-fixed, paraffin-embedded kidney biopsies of 14 patients with renal failure either clinically suspected for HFRS (7 cases) or caused by unknown (2 cases) or known other causes (drugs, sarcoidosis; 5 cases) were histologically investigated. An established S segment-specific PCR assay was applied to RNA isolated from the biopsies, and amplification products were verified by direct sequence determination. RESULTS: Investigations revealed a typical histopathological appearance for hantavirus infections in all seven suspected HFRS cases and one case of unknown cause. With five of the suspected HFRS cases, hantavirus-specific RNA was detected. Sequence comparison revealed a close relationship to corresponding nucleoproteins of known Puumala viruses. CONCLUSION: The established technique provides a simple and powerful tool that expands the diagnostic possibilities, especially for otherwise unidentified or retrospective cases. It further allows insight into the molecular epidemiology of HFRS-causing agents. (+info)
Consumer hazards of plastics.
(14/2410)
The modern consumer is exposed to a wide variety of plastic and rubber products in his day to day life: at home, work, school, shopping, recreation and play, and transport. A large variety of toxic sequellae have resulted from untoward exposures by many different routes: oral, dermal, inhalation, and parenteral. Toxic change may result from the plastic itself, migration of unbound components and additives, chemical decomposition or toxic pyrolysis products. The type of damage may involve acute poisoning, chronic organ damage, reproductive disorders, and carcinogenic, mutagenic and teratogenic episodes. Typical examples for all routes are cited along with the activites of Canadian regulatory agencies to reduce both the incidence and severity of plastic-induced disease. (+info)
Detection of translocation t(11;14)(q13;q32) in mantle cell lymphoma by fluorescence in situ hybridization.
(15/2410)
To assess an unequivocal diagnosis of mantle cell lymphoma (MCL), we have developed a fluorescence in situ hybridization (FISH) assay, enabling the demonstration of t(11;14)(q13;q32) directly on pathological samples. We have first selected CCND1 and IGH probes encompassing the breakpoint regions on both chromosomes. Then, we have defined experimental conditions enabling us to obtain bright clear-cut signals in all of the samples, independently of the initial fixation conditions. We have analyzed single-cell suspensions from 26 formalin-fixed, paraffin-embedded MCL samples with this set of probes. In all cases, we have found a fusion signal (ie, a t(11;14)(q13;q32) translocation) in 14% to 99% of cells (median, 87%). So far, IGH-CCND1 fusions have been detected in all of the 51 MCL patients that we have analyzed by FISH (either on paraffin-embedded tumor samples or on peripheral blood samples). Regarding the low sensitivity of other techniques used to diagnose t(11;14)(q13;q32) (ie, 70% to 75% for cytogenetics and 50% to 60% for polymerase chain reaction), our FISH assay is by far the most sensitive technique. Moreover, because of the quality of the fluorescent signals and the rapidity of the experiment, this technique is widely applicable, even in routine cytogenetics or pathology laboratories. As MCL patients are usually refractory to standard therapy, an unambiguous diagnosis is needed to propose adapted therapeutic strategies, and this highly sensitive assay may be of great value for accurate diagnosis in difficult cases. (+info)
Mechanical properties of mesenteric arteries in diabetic rats: consequences of outward remodeling.
(16/2410)
Diabetes induces hemodynamic and biochemical changes that can influence mechanical properties of arteries. Structure and mechanics of mesenteric small arteries were investigated in rats with streptozotocin-induced diabetes (duration 7-9 wk). The external diameter of mesenteric artery branches was measured in control (n = 9) and diabetic (n = 7) Wistar Rp rats at baseline and during pressurization in situ (0-150 mmHg) under normal and passive smooth muscle conditions. Mean arterial pressure and mesenteric artery pressure were not significantly different. Baseline mesenteric artery diameter was larger in the diabetes-induced group (439 +/- 12 vs. 388 +/- 18 micrometers, P < 0.05). Media cross-sectional area of arteries from diabetic rats was not significantly increased (0.0149 +/- 0.0015 vs. 0.0122 +/- 0.0007 mm2). Cross-sectional compliance was significantly increased in diabetic rats at intraluminal pressures ranging from 25 to 75 mmHg (P < 0.005), whereas cross-sectional distensibility was not modified. Wall tension and circumferential wall stress were increased in diabetes. These results indicate that mesenteric small arteries of diabetic rats display eutrophic outward remodeling associated with increased wall tension and circumferential wall stress. (+info)