Role of Escherichia coli K capsular antigens during complement activation, C3 fixation, and opsonization.

Escherichia coli strains with K capsular polysaccharides are relatively resistant to phagocytosis by polymorphonuclear leukocytes, in contrast to E. coli strains without K antigens. This inhibition of phagocytosis is related to an impaired recognition of the K+ strains by the phagocytes due to ineffective opsonization. All five strains without K antigens were readily phagocytized after opsonization in 5% normal serum, compared with no uptake of the K+ strains. Evidence is presented that the decreased opsonization of the K+ strains in normal serum is caused by a low rate of complement activation of the strains, with subsequent absence of C3b fixation or C3d fixation or both to the cell wall of the bacteria. After removal of the K+ antigens by heating of a K+ E. coli strain, the strain was able to activate complement, to bind C3b or C3d or both, and to become opsonized. Complement was then activated via the classical and alternative pathways, which was comparable to the complement consumption by K- E. coli.     

Tunica media of aorta and pulmonary trunk in relation to internal calibres in transposition of great arteries,in aortic and pulmonary atresia and in normal hearts

Summary In a post mortem material of 17 cases of transposition of the great arteries (TGA) from patients with an age range from birth to two years and ten months after birth, the internal calibres of the great arteries and the ostia of the heart proved to be the same as in normal hearts. Furthermore, the media of the ascending aorta and pulmonary trunk showed no adaptation to the abnormal circulatory conditions in 15 cases of TGA with an age range from birth up to 51/2 months: in both great arteries the thickness of the tunica media and the packing density of its elastic fibres were the same as in normal hearts. However, adaptation of the tunica media of the pulmonary trunk to the abnormal circulatory conditions: increased media thickness, was found in the two remaining cases, older than 12 months.In 7 cases of pulmonary atresia (age from 1 day to 12 months) and in 9 cases of aortic atresia (age from 2 days to 37 days) the following observations were made. Vessels with reduced or absent function (ascending aorta in aortic atresia and pulmonary trunk in pulmonary atresia) showed a markedly different structure. In aortic atresia the internal calibre and thickness of the media of the ascending aorta were markedly reduced, whereas the packing density of the elastic fibres of the media remained the same as in normal hearts. In pulmonary atresia the pulmonary trunk showed large variations in internal calibre, whereas both media thickness and the packing density of its elastic fibres remained the same as in normal hearts. When the markedly enlarged single functional vessels (the pulmonary trunk in aortic atresia and the ascending aorta in pulmonary atresia) were compared no significant differences between their internal calibre, media thickness and the packing density of the elastic fibres were found indicating similar adaptation to the abnormal but comparable functional load of acting as sole arterial trunk.We are indebted to Prof. Dr. J. Moll for his help, to Dr. J.J. Willemse for statistical calculations, to Mr. P. Zondervan, M.D. (Dept. of Pathology I), for supply of material and to Mrs. L. Silvis for histo-technical assistance     

CD1 assembly and the formation of CD1-antigen complexes

The CD1 antigen presentation system presents lipid antigens to effector T cells, which have diverse roles in antimicrobial responses, antitumor immunity and in regulating the balance between tolerance and autoimmunity. The trafficking of CD1 molecules and lipid antigens facilitates their intersection and binding in specific intracellular compartments. Recent studies have now identified unexpected accessory molecules that are critical to CD1 assembly and lipid loading. The atomic structures of CD1-antigen complexes have defined both the orientation of polar headgroups between the alpha1 and alpha2 helices of CD1 and the manner in which distinct CD1 isoforms bind a range of lipids that have different lengths and numbers of hydrocarbon chains.     

Chicks from a high and low feather pecking line of laying hens differ in apomorphine sensitivity

Proactive rodents show a larger behavioral response to apomorphine (APO) than reactive copers, suggesting a more sensitive DA system in proactive individuals. Previously, chicks from a high feather pecking (HFP) and low feather pecking line (LFP) have been suggested to display a proactive and reactive coping strategy, respectively. Therefore, at approximately 4 weeks of age, the behavior of 48 LFP and 48 HFP chicks in response to an APO injection was studied using an open field. Another objective of the present study was to determine whether behavioral variation (in an open field) between HFP and LFP birds, after APO injection, is also reflected by variation of D(1) and D(2) receptor densities in the brain. Receptor binding capacities were assessed by measuring specific binding of tritiated D(1) and D(2) receptor ligands in different regions of the brain of control HFP and LFP chicks. In the present study, it is shown that indeed HFP chicks display a more enhanced behavioral response to acute APO treatment (0.5 mg/kg BW) than LFP birds in an open field. This difference was not reflected by variation of D(1) and D(2) receptor densities in the brain between both lines.     

Capacitation-dependent concentration of lipid rafts in the apical ridge head area of porcine sperm cells

Lipid architecture of the plasma membrane plays an important role in the capacitation process of the sperm cell. During this process, an increase in membrane fluidity takes place, which coincides with a redistribution of cholesterol to the apical region of the head plasma membrane and subsequently an efflux of cholesterol. Cholesterol is also a major player in the formation of lipid rafts or microdomains in the membrane. Lipid rafts favour specific protein-protein interactions by concentrating certain proteins in these microdomains while excluding others. In this study, we investigated the organization of lipid rafts during in vitro capacitation of boar sperm cells. We report on the presence of the lipid raft-specific proteins caveolin-1 and flotillin-1 in sperm cells. Capacitation induced a change in membrane distribution of these proteins. Lipid analysis on detergent-resistant membranes (DRMs) of sperm cells indicated that capacitation induces a lipid raft concentration rather than a disintegration of lipid rafts, because the total amount of lipid in the DRM fraction remained unaltered. Using a proteomic approach, we identified several major DRM proteins, including proteins involved in capacitation-dependent processes and zona pellucida binding. Our data indicate that sperm raft reorganization may facilitate capacitation-specific signalling events and binding to the zona pellucida.     

Effect of tissue fixatives on telomere length determination by quantitative PCR

Telomere length is a well established marker of cellular senescence and thus biological age. Quantitative PCR allows the determination even from very low amounts of tissue by using telomere specific and single copy gene primers. Comparing a directly processed tissue sample to a 4% formaldehyde fixed one showed a significantly reduced efficiency of PCR reactions (mainly in single copy gene experiments) in a storage time-dependent manner resulting in an artificial increase in reported relative telomere length. This effect was not seen when the tissue was stored in RNA later solution. In summary, telomere length determination from formaldehyde fixed material by quantitative PCR is not a reliable method. Unfortunately therefore, many easily accessible tissue samples from pathology laboratories are unsuitable for this technique.     

Use of two parallel oxygenators in a very large patient (2.76 m2) for an acute "A" dissecting aortic aneurysm repair

The very large patient (weight 142 kg, height 197 cm, body surface 2.76 m2) was referred to acute operation with dissecting type A ascending aortic aneurysm. The calculated blood flow was 6.63 l/min. To anticipate potential difficulties with perfusion and oxygenation two oxygenators connected in parallel were incorporated into the circuit. Bentall procedure with ACB to the RCA was performed. The perfusion was uneventful. Bypass time was 259 minutes, cross clamp time 141 minutes, circulatory arrest 7 minutes. The highest oxygenators gas flow was 2.6 l/min with maximum FiO2 0.42. The use of two in parallel connected oxygenators is a very effective, easy and safe method in such extreme perfusions, offering to the perfusionist a great reserve of oxygenator output.     

Caspase-dependent and -independent DNA fragmentation in Sertoli and germ cells from men with primary testicular failure: relationship with histological diagnosis

BACKGROUND: Germ cell elimination and sperm DNA fragmentationin men with primary testiculopathies involve apoptosis-relatedprocesses whose mechanisms are poorly understood. This studyexamines the participation of typical (caspase-dependent) andatypical (caspase-independent) pathways in these processes.METHODS: Caspase activity and DNA fragmentation were evaluatedin Sertoli and germ cells from 63 men with non-obstructive azoospermiaand with different histological diagnoses who were undergoingtesticular biopsy for an assisted reproduction attempt. In eightof these men, phosphatidylserine externalization was also examined.RESULTS: The percentage of Sertoli cells showing caspase activityand DNA fragmentation was low and uniform in all diagnoses.In germ cells that remained tightly associated with Sertolicells despite vigorous mechanical treatment, the incidence ofboth caspase activity and DNA fragmentation was high, particularlyin men with maturation arrest. In Sertoli cell-free germ cells,high incidence of DNA fragmentation contrasted with low incidenceof caspase activity and phosphatidylserine externalization.CONCLUSIONS: In men with primary testicular failure, apoptosisof Sertoli cells is insignificant. Some germ cells undergo caspase-dependentapoptosis, show phosphatidylserine externalization and are tightlyassociated with Sertoli cells. Other germ cells show caspase-independentDNA fragmentation, do not externalize phosphatidylserine andlack a tight association with Sertoli cells.