Protection to Fasciola hepatica in the gut mucosa of immune rats is associated with infiltrates of eosinophils,IgG1 and IgG2a antibodies around the parasites

We investigated the immune effector mechanisms that underlie protection against F. hepatica in the gut wall of immune rats, using (immuno)histochemistry. In the lamina propria of immune Wistar rats, four weeks after oral infection, frequencies of IgE-positive cells, eosinophils and mucosal mast cells were significantly increased, compared with naïve rats. These factors represent the traditional effector mechanisms against helminths. No significant differences were detected between the two groups in frequencies of IgM-, IgG2a-, IgG1- and IgA- positive cells, CD4- and CD8-positive cells, NK cells, macrophages, neutrophils or goblet cells. Upon challenge of immune rats with F. hepatica in an ex vivo gut segment, NEJs that migrated through the (sub)mucosa were coated with IgG1 and IgG2a antibodies and surrounded by eosinophils. No IgE or IgA antibodies were detected on the parasites. The onset of these immune effector responses, two h after challenge, was related to the expression of protection. These results suggest that NEJs are killed by an eosinophil-mediated cytotoxic response involving IgG antibodies. These antibodies were not produced in the intestine, but infiltrated the gut upon challenge. The observed immune effector responses were not restricted to the site where the primary infection is located, namely the small intestine, but were also detected in the large intestine. The presence of the protective immune mechanisms in two other rat strains demonstrates the pivotal importance of these responses, irrespective the genetic background of the host.     

Protective immunity induced by vaccination with two Haemonchus contortus excretory secretory proteins in sheep

Two excretory secretory (ES) antigens of adult Haemonchus contortus with molecular weights of 15 and 24 kDa, respectively, were evaluated as protective immunogen against haemonchosis. Sheep were vaccinated three times and subsequently challenged with 20 000 infective larvae. Vaccination induced significant reduction (>70%) in mean faecal egg counts and abomasal worm burden compared to the non-vaccinated control group or adjuvant control group. Vaccination induced ES-specific antibodies and stimulated infiltration of mast cells in the abomasal tissue.     

Localized sinus inflammation in a rabbit sinusitis model induced by Bacteroides fragilis is accompanied by rigorous immune responses

We evaluated inflammatory and immune responses against Bacteroides fragilis in a rabbit sinusitis model. Bacteroides was inoculated into the left maxillary sinus, and inflammatory (histology, cell number/cytology, lactose dehydrogenase, and apoptosis) and immune responses in the sinus, airway, and peripheral blood (PB) were determined for up to 4 weeks. In the inflamed sinus, the lactose dehydrogenase level was markedly elevated, with neutrophilic infiltration, severe tissue inflammation, and increased apoptosis. Low-grade tissue inflammation was present in the contralateral and sham-operated sinuses, but other parameters remained unchanged, and so did those in the airway and PB in the inoculated rabbits. Serum IgG antibody levels increased rapidly, were highest at 3 weeks, and began to decline at 4 weeks. Cellular immune responses (proliferation and interferon-gamma mRNA expression) against Bacteroides were detected in the PB of all inoculated rabbits. Vigorous immune responses against Bacteroides may have localized but failed to terminate inflammation in the sinus, indicating importance of microenvironmental factors.     

Economic evaluation in clinical nephrology: Part 1. An introduction to conducting an economic evaluation in clinical practice

Decision‐making in clinical practice is complex and getting more complex. There are a large range of alternative actions possible, all with different consequences and trade‐offs. The complexity of medical decision‐making is best illustrated using a clinical scenario.     

Ca2+ involvement in the action potential generation of myenteric neurones in the rat oesophagus

Intracellular recordings were used to study the physiological behaviour of rat oesophageal myenteric neurones, which are embedded in striated muscle. Injection of depolarizing pulses evoked action potentials with a clear 'shoulder' in all neurones. This shoulder disappeared under low Ca2+/high Mg2+ conditions. Tetrodotoxin (TTX; 1 micromol L-1) did not impede spike firing, whereas under combined TTX and low Ca2+/high Mg2+ conditions the action potentials were completely abolished, indicating that TTX- resistant action potentials are mediated by a Ca2+ current. Further experiments with omega-conotoxin GVIA (100 nmol L-1) revealed that these Ca2+ currents enter the cell via N-type voltage-activated Ca2+ channels (see also accompanying paper). Tetraethylammonium (10 mmol L-1) caused broadening of the action potentials, which probably resulted from prolonged Ca2+ influx due to blockade of the delayed rectifier K+ channel. Although Ca2+ appears to be involved in the spike generation of all rat oesophageal myenteric neurones, only a minority (14%) shows a slow afterhyperpolarization. Thus, no strict correlation exists between the presence of a shoulder and a slow afterhyperpolarization. Furthermore, morphological identification of 25 of the impaled neurones revealed that there was no strict correlation between morphology and electrophysiological behaviour. Consequently, rat oesophageal myenteric neurones appear to differ in several aspects from myenteric neurones in smooth muscle regions of the gastrointestinal tract.     

Synergism of the antimicrobial agents miconazole and benzoylperoxide

Miconazole was active against aerobically grown Staphylococcus spp., and anaerobically grown Propionibacterium acnes. The combination of this N-substituted imidazole derivative with benzoylperoxide resulted in a marked increase in bactericidal activity. The antibacterial activity observed in vitro with the combination is greater than the sum of the effects observed with the two drugs independently, indicating bactericidal synergism.     

Electrophysiology of porcine myenteric neurons revealed after vital staining of their cell bodies. A preliminary report

Abstract Due to practical limitations in visualizing and getting access to the ganglionic components of large mammals, electrophysiology of the enteric nervous system has been restricted mainly to small laboratory animals, more particularly the guinea-pig. The use of the vital dye 4-(4-diethylaminostyryl)-N-methylpyridinium iodide (4-Di-2-ASP), however, overcomes some of these difficulties. A 20-min incubation period with this dye, followed by a minimum period of 4 h in Krebs solution, suffices to stain the neuronal cell bodies, permitting selection of a neuron and positioning of the microelectrode for impalement and recording. The method has been applied to pig ileum and guinea-pig large and small bowel myenteric neurons. Impalements of untreated guinea-pig myenteric neurons were compared with those of 4-Di-2-ASP-pre-treated ones. According to our preliminary data, the staining did not suppress the expression of apparently normal electrophysiological activity. Moreover, the procedure permitted impalement and recording of myenteric plexus neurons in pig ileal tissue with a rate of success equalling blind impalement on guineapig tissue. In contrast with formerly published results whereby staining of the neuronal cell bodies only occurred when the cells had been chemically damaged, our experiments suggest a possible correlation between fluorescence and cell viability.     

Protection against Fasciola hepatica in the intestine is highly correlated with eosinophil and immunoglobulin G1 responses against newly excysted juveniles

Rats were infected with Fasciola hepatica and challenged at regular intervals up to 38 weeks using an ex vivo gut loop, a technique developed in our laboratory. The kinetics of the observed immune responses against F. hepatica in gut tissue and serum were investigated and correlated to protection. Immunohistochemical methods were used to measure the frequency of eosinophils, immunoglobulin (Ig)E-positive cells, and mucosal mast cells in the gut loop, and to determine whether the newly excysted juveniles were coated with IgG antibodies or surrounded by eosinophils, or both. Enzyme-linked immunosorbent assays and a radioimmuno assay were used to measure serum antibody reactive with newly excysted juveniles. Results showed that protection was highly correlated with the frequency of eosinophils and IgE-positive cells in the gut, but was only moderately correlated with the frequency of mucosal mast cells. Newly excysted juveniles taken from rats exhibiting high levels of protection were always coated with IgG antibodies and surrounded by eosinophils. Protection was highly correlated with titers of serum IgG1 antibodies directed against newly excysted juveniles, but was only weakly correlated with titers of serum IgA and IgE antibodies. Because protection was highly correlated with IgG1 in gut tissue and serum, and with eosinophils in gut tissue, we suggest that IgG1 and eosinophils are important in protecting rats against F. hepatica.     

Circadian Characteristics of Urinary Melatonin from Clinically Healthy Young Women at Different Civilization Disease Risks

ABSTRACT Rhythm characteristics in the about-daily (circadian) and about-yearly (circannual) frequency ranges were assessed for urinary melatonin. Clinically healthy women in Minnesota, USA, and Kyushu, Japan, were sampled around the clock once in 1-4 seasons. Possible differences that could reflect the large difference in breast cancer incidence in these two geographic locations were investigated. Each subject's risk of developing breast cancer, cardiovascular diseases resulting from an elevated blood pressure, and emotional conditions was numerically evaluated according to epidemiologic questionnaires. A prominent circadian rhythm characterizes urinary melatonin in both populations, peaking in the middle of the night. The American women exhibit a larger circadian rhythm-adjusted mean (mesor) than do the Japanese women. A circannual rhythm is also apparent in the North American women, but not in the Japanese women. The circadian mesor of urinary melatonin correlates negatively with the risk score of emotional depression and positively with that of developing cardiovascular diseases.