Workshop on a detailed characterization of Trichinella spiralis antigens: a platform for future studies on antigens and antibodies to this parasite

In order to characterize immunodominant components of T. spiralis a workshop was organized. In this the reactivity of monoclonal and polyclonal antibodies, provided by different research groups, towards total extracts from adult, new born larvae and muscle larvae as well as to excretory/secretory components of muscle larvae were tested by ELISA, Western blot and immunoprecipitation assays. As a result of this workshop T. spiralis ML antigens have been classified into eight groups (TSL-1-TSL-8) according to their recognition by monoclonal and polyclonal antibodies. Among them, TSL-1 antigens have been the most extensively characterized both biochemically and immunologically. These antigens are stage specific, originate in the muscle stichosome and are abundant in both E/S and on the larval cuticular surface. The TSL-1 antigens share an immunodominant carbohydrate epitope (tyvelose), which is unique for Trichinella and is not associated with phosphorylcholine. The data collected in this workshop has allowed both the unification of the nomenclature for T. spiralis antigens and their biochemical characterization. It also has provided a platform for further studies on the characterization of other T. spiralis antigens and indeed for other Trichinella species.     

Clostridium perfringens invasiveness is enhanced by effects of theta toxin upon PMNL structure and function: the roles of leukocytotoxicity and expression of CD11/CD18 adherence glycoprotein

Clostridium perfringens infections are characterized by the lack of an inflammatory response at the site of infection and rapidly progressive margins of tissue necrosis. Studies presented here investigated the role of theta toxin from C. perfringens in the pathophysiology of these events. Mice passively immunized with neutralizing monoclonal antibody against theta toxin and challenged with an LD100 of log phase C. perfringens had significantly less mortality than untreated controls. Intramuscular injection of killed, washed C. perfringens in mice induced a massive time-dependent influx of polymorphonuclear leukocytes (PMNL) into tissue; injection of either viable, washed C. perfringens or killed organisms plus theta toxin dramatically attenuated PMNL influx although PMNL accumulated in adjacent vessels. The anti-inflammatory effects could not be attributed to an absence of chemoattractants since C. perfringens proteins had chemotactic factor activity, and killed bacilli generated serum-derived chemotactic factors. Scanning and transmission electron microscopy demonstrated the dramatic leukocidal effects of high doses of theta toxin on PMNL. In contrast, sublethal concentrations of theta toxin primed PMNL chemiluminescence, disrupted PMNL cytoskeletal actin polymerization/disassembly, and stimulated functional upregulation of CD11b/CD18 adherence glycoprotein. In summary, these results demonstrate that theta toxin is an important virulence factor in C. perfringens infection. In a concentration-dependent fashion, theta toxin contributes to the pathogenesis of clostridial gangrene by direct destruction of host inflammatory cells and tissues, and by promoting dysregulated PMNL/endothelial cell adhesive interactions.     

Coronary flow analysis during autoperfusion angioplasty

BACKGROUND: Autoperfusion balloons are available for the protection of the myocardium during balloon angioplasty. The aortic pressure is the driving force that delivers blood to the distal vessel during balloon inflation. Autoperfusion balloons can achieve sufficient flow rates in vitro. The use of these devices is recommended in high-risk patients in danger of haemodynamic collapse during balloon inflation. The quantity of the distal blood flow during balloon inflation in vivo is still unknown. OBJECTIVES: To measure distal coronary perfusion using Doppler guidewires during percutaneous transluminal coronary angioplasty (PTCA) with autoperfusion balloons. METHODS: Coronary flow velocity was measured with 0.014-inch Doppler guidewires bypassing the autoperfusion balloon in eight patients undergoing elective PTCA (degree of stenosis 74 +/- 7.2%). We used balloons with diameters of 3.0 and 3.5 mm. The coronary diameter at the location of the flow measurements was obtained by quantitative angiography in two planes. Coronary blood flow was calculated as the luminal area multiplied by the average peak flow velocity of the Doppler wire divided by 2. Coronary flow velocity reserve was measured before and after angioplasty by intracoronary injection of adenosine. RESULTS: Coronary blood flow was 35 +/- 11.6 ml/min before PTCA. During average inflation times of 4.6 +/- 0.9 min, coronary blood flow was 19 +/- 3.8 ml/min (P = 0.002) after withdrawing the guidewire in the autoperfusion balloon. Five minutes after angioplasty it increased to 42 +/- 13.5 ml/min (P < 0.001). Four patients had electrocardiographic changes during balloon inflation; three patients reported chest pain. One patient required a stent because of a local dissection. To achieve satisfactory angiographic results (residual stenosis 11 +/- 8.5%), we performed 2.1 +/- 0.78 inflations on average with a cumulative inflation time of 8.8 +/- 3.35 min. Coronary flow velocity reserve increased from 1.3 +/- 0.20 to 2.2 +/- 0.22 (P < 0.001). CONCLUSIONS: Using the autoperfusion balloon we measured a coronary blood flow during angioplasty of 56 +/- 10.3% of the distal perfusion before PTCA. In high-risk patients dependent on adequate coronary perfusion, autoperfusion balloons are not able to provide sufficient distal coronary blood flow during balloon inflation. In these patients active coronary or circulatory support devices are recommended.     

Weight gain as an indicator of response to chemotherapy for oesophageal carcinoma

Patients with oesophageal carcinoma commonly present with dysphagia and weight loss, which may be related to the tumour burden and/or the physical obstruction to the passage of food. In this study we have examined the relationship between weight change and response to chemotherapy in 28 patients undergoing neo-adjuvant chemotherapy for squamous or anaplastic carcinoma. Two pulses of mitomycin, ifosfamide and cisplatin were given 3 weeks apart. Body weights were measured prior to the first pulse and 3 weeks after the second. Patients underwent oesophageal dilatation routinely at diagnostic endoscopy prior to chemotherapy, in order to permit oral nutrition. No dietary modifications were made. Tumour response was assessed on a barium swallow. Patients had a normal spread of weights on presentation. In the non-responding group (n = 9), eight patients lost weight and one gained weight. Of the partial responders (> 50% tumour shrinkage; n = 11), five gained weight, five lost weight and one remained constant. In the complete response group (n = 8), six gained weight and two lost weight. Statistical analysis showed a significant difference (F = 4.61; P = 0.02) between change in weight expressed as a percentage of ideal weight in nonresponders (mean -5.3%) versus partial responders (mean +2.4%), and in non-responders versus complete responders (mean +1.1%). Weight gain during chemotherapy is a good indication of response, although its absence does not preclude a response. In the majority of patients who respond to chemotherapy there will be an increase in weight with improvement in their general condition prior to operation.     

Intrinsic differences in various segments of the proximal convoluted tubule

Until recently it has not been possible to compare directly the function of superficial and juxtamedullary nephrons. The present studies, using in vitro microperfusion, were designed to examine whether functional differences exist between proximal convoluted tubule segments of superficial and juxtamedullary nephrons. Electrophysiological studies showed that major differences exist between the relative chloride and sodium permeabilities of these segments. In the 1st mm of the superficial proximal convoluted tubule, the permeability to sodium was greater than that to chloride, whereas in the 2nd mm of the superficial proximal convoluted tubule and all later segments, the permeability to chloride was greater than that to sodium. The juxtamedullary proximal convoluted tubule was found to differ from the superficial proximal convoluted tubule in two respects: first, the relative permeabilities to chloride and sodium did not differ in the various segments of the juxtamedullary proximal convoluted tubule; second, the permeability to sodium was greater than to chloride throughout. When perfused with a solution lacking glucose and amino acids, the superficial and juxtamedullary convolutions exhibited the same transepithelial potential change, a reversible decrease to less than -- 1 mV. It thus appears that in both convolutions there exists electrogenic sodium transport coupled to the transport of these organic solutes. This differs from pars recta of both of these nephrons, which have been shown to exhibit electrogenic sodium transport independent of organic solutes. However, when perfused with a solution lacking glucose and amino acids but also containing high chloride and low bicarbonate concentrations, the superficial convolution developed a significantly more positive potential than the juxtamedullary. This difference reflects greater relative chloride permeability in the superficial proximal convolution. These studies show that intrinsic functional differences exist between proximal convoluted tubules obtained from the superficial and juxtamedullary nephron populations.