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Biomedical subjects

M Stankiewicz

Publications and source records attributed to M Stankiewicz.

At least 55 records · Page 3Linked to original sources
PubMed1996

Drug-abbreviated infections and development of immunity against Trichostrongylus colubriformis in sheep.

A protective immune response without liveweight loss can be induced in sheep against T. colubriformis but results depend on the anthelmintic used and duration of immunizing infections. More than 90% protection was achieved in sheep immunized by three 15- or 7-day oxfendazole abbreviated infections or three 21-day nonabbreviated infections. Only 41% protection was induced by 3-day oxfendazole abbreviated infections. Significantly higher worm burden and faecal egg counts were present after challenge in sheep immunized by 7-day levamizole abbreviated infections compared to 7-day oxfendazole abbreviated infection. Liveweight gains of sheep immunized by 15- and 7-day abbreviated infections were not significantly different than non infected controls. Liveweight loss seemed to be associated with high activity of mucus peroxidase and high numbers of eosinophils in the intestinal lumen. High parasite numbers seemed to be associated with low activity of alkaline phosphatase in mucus. Mucus peroxidase, arylsulphatase, larval migration inhibition of mucus, mucus or serum antibody against L3 excretory/secretory antigen or somatic L3, L4 and adult antigen were not associated with protection.

Alkaline Phosphatase↗
PubMed1996

Supravital staining of eosinophils.

Live eosinophils when mixed with Acridine Orange solution and viewed microscopically using u.v. light show very intense colours of their granules (yellow, orange and red) and green nuclear staining. Their active movement, translocation of granules and degranulation can be observed in vitro. Using this method, live eosinophils can be easily differentiated and enumerated.

Acridine Orange↗
PubMed1996

The immune response of sheep surgically modified with intestinal loops to challenge with Trichostrongylus colubriformis.

Aspects of the local immune response to nematode challenge were investigated in vivo in isolated loops of the upper small intestine of mature sheep that were immunised by repeated infections with Trichostrongylus colubriformis infective larvae (L3). Groups of 3 sheep were challenged either through the loop (Group 1) or orally (Group 2) with T. colubriformis L3, the third group served as unchallenged controls (Group 3). Nematode specific antibody levels, mast cell proteinase levels (SMCP) and larval migration inhibition (LMI) activity were determined in loop secretions for 4 weeks after challenge. The intestinal loops remained functional throughout the experiment. Groups 1 and 2 were re-challenged 2 weeks after the first challenge, and all 3 Groups were slaughtered 2 weeks later. Histopathological examination showed elevated numbers of globule leukocytes (GL) in both the nematode-challenged loop and unchallenged small intestine of Group 1 and small intestine of Group 2 indicating that nematode infections induce the local appearance of large numbers of GL. Oral, but not loop challenge caused increased antibody levels in loop secretions when compared to unchallenged controls. Only loop-challenged sheep showed a peak in loop fluid SMCP levels 10-13 days after the first challenge which coincided with a peak in numbers of mucosal GL. The isolated loops of all 3 groups showed highly elevated numbers of eosinophils when compared to the intact small intestine. Loop fluid of all 3 groups showed a high level of LMI activity reflecting the high level of nematode-resistance induced by the immunisation procedures. Sheep in Groups 1 and 2 were both able to expel challenge infections, and when compared to Group 3, showed higher blastogenic activity of unstimulated cells derived from a mesenteric lymph node in the region of the challenged part of the intestine. The present experiment showed that surgically constructed intestinal loops provided a model system by which the substantial changes associated with the local intestinal immune response to challenge with T. colubriformis could be investigated.

Animals↗
PubMed1996

Immune responsiveness of nematode-resistant or susceptible Romney line-bred sheep to continuous infection with Trichostrongylus axei.

Two divergent lines of Romney sheep have been selected on the basis of differences in faecal egg counts (FEC) to natural poly-generic parasite challenge in New Zealand. However, it is not known if the expression of resistance or susceptibility extends to parasitic nematodes that were not a major part of the selection challenge. To examine this, the immune response to infection with Trichostrongylus axei was examined in these sheep lines. Changes in the proportions of CD5+, CD4+, CD8+, T19+ and CD45R+ lymphocytes and parasite specific antibody titres in peripheral blood were monitored each week in six resistant and six susceptible line lambs that were maintained indoors in pens during the course of 14 weekly infections with 10,000 T. axei larvae. No difference in FEC was observed. Similarly, no significant difference in T. axei specific antibody titre between sheep lines was seen although antibody titre increased steadily from Week 4. Significant increases in the proportions of CD5+, CD4+, CD8+ and T19+ cells occurred in both resistant and susceptible line lambs during Weeks 1-4 of infection. Following peak levels, proportions of CD5+, CD4+ and CD8+ cells fell with the rate of decline of CD5+ and CD4+ cells significantly greater in the resistant line lambs. Proportions of CD45R+ cells showed significant changes with time that were the inverse of those of CD5+, CD4+ and CD8+ cells. Susceptible line lambs showed higher proportions of CD5+ and lower proportions of CD45R+ cells than resistant lambs before infection with T. axei. Overall, during infection, these differences were maintained while CD4+ and T19+ levels were higher in the susceptible line lambs.

Analysis of Variance↗
PubMed1996

Field studies of the immunisation of lambs with drug-abbreviated infections of Trichostrongylus colubriformis and Ostertagia circumcincta.

Fifty, 5-6 month-old lambs were randomly allocated to five equal groups. Three groups of lambs were immunised by three oxfendazole-abbreviated (artificial) infections of Trichostrongylus colubriformis and Ostertagia circumcincta. Group 1 was immunised with a high dose of larvae, Group 2 with a medium dose and Group 3 with a low dose. A fourth group was treated with oxfendazole only and a fifth group was not treated (control). All groups were grazed together on pasture naturally contaminated with nematode larvae. Immunisation significantly reduced the number of eggs per gram of faeces in all three groups, but the lowest faecal egg count was seen in Group 3 (immunised with the lowest number of larvae). Significantly better liveweight gains and wool weight were observed in Group 2 animals than in the control group. Of all anthelmintic-treated animals, only Group 1 lambs did not have a significantly lower dag weight than controls.

Journal Article↗
PubMed1996

Effect of K+ accumulation removal and high extracellular osmolarity on K+ current in insect axonal membrane.

The effect of a removal of K+ accumulation on K+ current in insect axonal membrane was observed. Experiments were performed on isolated giant axon of a cockroach using double oil gap technique. K+ accumulation was reduced by: (1) an outward water flow induced by non-electrolytes (urea, glucose) added to extracellular saline and (2) by an increase of non-specific permeability of axonal glial layer obtained after the application of DMSO. The conclusions are: (1) osmolar effect depends on the type of molecule used for osmotic shock, (2) increase of outward K+ current in conditions of high extraaxonal osmotic pressure is attributed to the decrease of K+ accumulation and outward water flow, (3) removal of K+ accumulation doesn't affect the kinetics and the time course of K+ current, (4) experiments confirmed the presence of an inactivating component in the axonal outward K+ current, (5) DMSO must be used cautiously as a solvent in electrophysiological experiments.

Animals↗
PubMed1996

[Chlamydia trachomatis and concomitant flora in the internal genital organs of women without clinical symptoms of infection].

One thousand and forty swabs from 52 gynaecological patients were examined bacteriologically. The most frequently isolated pathogen was C. trachomatis found in the endocervix in 19.2%, in the endometrium in 17.3% and in the fallopian tube in 48.1% of the examined women. There was also shown that presence of C. trachomatis in utero and in fallopian tube not always coexist with it's presence in the endocervix, move over presence of that microbe doesn't have influence on presence of any bacteria in genitourinary tract.

Bacteria↗
PubMed1995

Levamisole and its influence on the immune response of lambs.

Ten parasite-free lambs were drenched with 8 mg/kg of levamisole on days 0 and 28 and were injected with human erythrocytes and ovalbumin one day after each drench. Ten other antigen-injected lambs were not drenched with anthelmintic as controls. Lymphocytes from the control and drenched lambs were cultured in vitro with RPMI 1640 plus 5% fetal calf serum (FCS), with 50% autologous serum only, with concanavalin A (Con A) or with phytohaemagglutinin (PHA). Decreased blastogenesis was observed in cells from the drenched lambs cultured in the presence or absence of mitogen and was most obvious when 50% autologous serum was used, particularly with PHA, and when lymphocytes were collected 3 and 7 days after the first and 3 days after the second antigen injection. There were no significant changes in antibody titres between the groups. Decreased serum complement activity was seen 3 days after the second antigen injection in the drenched lambs. Although there was a significant reduction in the serum insulin-like growth factor I levels 4 days after each levamisole drench, the drenched lambs gained significantly more weight than the non-drenched control lambs.

Animals↗
PubMed1995

Immunization of sheep against parasitic nematodes leads to elevated levels of globule leukocytes in the small intestine lumen.

In sheep that had been given three immunizing infections with Trichostrongylus colubriformis and Ostertagia circumcincta infective (L3) larvae, drenched after the last infection and challenged with larvae of the same species, there was a significant increase in numbers of small intestine mucosal tissue globule leukocytes (TGLs) and lumenal globule leukocytes (LuGLs) compared with sheep that had only been drenched and challenged. There was a positive correlation between the numbers of LuGLs and TGLs in the small intestine but the ratio of these two cell types was lower in non-immunized than immunized sheep. In immunized sheep positive correlations were observed between LuGLs and levels of arylsulphatase and peroxidase in the intestinal mucus and between arylsulphatase and larval migration inhibition (LMI) activity in mucus. Lumen eosinophils correlated with blood eosinophils, serum antibody against T. colubriformis correlated with peroxidase in the mucus and blood eosinophils correlated with nematode specific IgM levels in the intestinal mucus. In the abomasum, TGLs were present but not LuGLs. Sheep repeatedly infected with T. axei also had significantly more LuGLs in the small intestine than control animals. Two sheep that had a surgically prepared isolated small intestinal loop, after oral infection with T. colubriformis had TGLs and LuGLs in the intact intestine, but not in the isolated loop. Significantly more LuGLs were produced in sheep by allowing repeated T. colubriformis L3 infections to develop to adult stages compared to sheep treated with the same number of larvae, but where the infections were terminated by drenching at various intervals.

Animals↗
PubMed1995

The immune responsiveness of Romney sheep selected for resistance or susceptibility to gastrointestinal nematodes: lymphocyte blastogenic activity, eosinophilia and total white blood cell counts.

Blastogenic activity, eosinophil and total white blood cell counts (TWBC) were examined over a period of 14 weeks in Romney lambs, genetically resistant or susceptible to gastrointestinal nematodes. The lambs were infected with 5000 infective Trichostrongylus colubriformis larvae twice weekly. Compared to preinfection levels, the blastogenic activity of unstimulated lymphocytes in lambs of both lines peaked at week 3, and was significantly higher in resistant than in susceptible lambs. These changes may have been due to in vivo polyclonal activation. Lymphocytes from susceptible sheep responded more strongly to Con A, PHA and PWM than cells from resistant sheep. Counts per minute (c.p.m) for Con A- and PHA-stimulated lymphocytes increased in both lines of sheep from week 2 to week 7 and then returned to initial levels. An increase in c.p.m. in PWM-stimulated cell cultures was observed from weeks 3 to 5 in both groups. The blastogenic activity for LPS-stimulated cultures was significantly higher for resistant than susceptible sheep at weeks 3 and 4. No significant correlations between the decline in faecal egg counts (FEC) and the blastogenic activity was observed. Eosinophil counts in peripheral blood began to increase one week earlier in resistant than in susceptible sheep. No significant correlation between FEC and eosinophil counts was observed in resistant lambs, whereas in susceptible lambs a significant correlation was found between FEC and eosinophil counts at some sampling times. TWBC in resistant lambs steadily increased with infections whereas susceptible lambs showed a decrease until week 5 and then steadily increased. There was no significant correlation between the decline in FEC and TWBC.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Influence of ivermectin on cellular and humoral immune responses of lambs.

In view of the extensive use of anthelmintics in sheep and the fact that their activity may in part depend upon the immune system, we were interested to determine if ivermectin had any influence on aspects of the sheep immune response. Ten parasite-free 6-month-old lambs were drenched with ivermectin and 1 day later were given intravenously human erythrocytes and subcutaneously ovalbumin. Ten other lambs with injected antigens were not drenched and served as controls. Both groups were bled at intervals for cells and serum. The procedure was repeated on day 28. Lymphocytes from the drenched lambs, cultured in vitro in RPMI plus 50% autologous serum collected up to 7 and 14 days after the first and second antigen injections respectively, had decreased blastogenic activity compared with lymphocytes from control lambs. Similar results were obtained with lymphocytes cultured in RPMI 1640 supplemented with 50% autologous serum plus concanavalin A (Con A) or phytohaemagglutinin (PHA). When washed, lymphocytes were cultured in RPMI 1640 supplemented with 5% foetal calf serum (FCS) or 5% FCS plus Con A or PHA, decreased blastogenesis was observed but blastogenesis depression was not as marked as that observed with autologous serum. Similar antibody responses were seen for the drenched and control groups in response to the two injections of both antigens except that after the second injection, there was a significant reduction in antibody response to ovalbumin in the ivermectin-treated lambs. There were no differences in serum complement or serum nitric oxide levels between the two groups at any stage, but insulin-like growth factor-1 levels were significantly reduced in serum of the ivermectin-treated group, 4 days after each drench. Growth hormone levels were consistently significantly higher 22 days after both drenchings. There was no difference in mean body weight increase between the groups during the experiment.

Animals↗
PubMed1995

Immune responsiveness of Romney sheep selected for resistance or susceptibility to gastrointestinal nematodes: field studies.

Long-term selection of sheep for resistance to parasite infections may be jeopardized if animals do not retain their normal ability to respond to non-parasite antigens. Therefore the antibody responses to ovalbumin (OVA) and human red blood cells (HRBC), and kinetics of peripheral blood lymphocyte phenotypes were examined in mature grazing sheep, genetically resistant or susceptible to gastrointestinal nematodes. In both lines the HRBC antibody response peaked 2 weeks after the primary injection, 1 week after the second injection and 3 weeks after the second OVA injection. The antibody titres of the resistant line sheep decreased sooner after both primary and secondary injections. The resistant line sheep had higher percentages of CD5+ and CD4+ cells than the susceptible sheep. Two injections of OVA and HRBC did not result in significant alterations in percentages of CD5+, CD4+, CD8+ and CD45R+ lymphocytes in either line. In both lines, the control groups showed a steady increase of 0.29% per week in percentages of T19+ (gamma delta) T cells which was significantly higher than in the antigen injected sheep.

Animals↗
PubMed1995

Effects of repeated systemic penicillin injections on nonconvulsive and convulsive epileptic seizures in the rat.

Changes in the spontaneous and induced epileptic activity in the course of repetitive systemic i.p. administration of crystalline penicillin (Pc) were examined in imp-DAK rats. In all the rats used, nonconvulsive seizures characterized by bursts of spike and wave discharges (SWD) in the neocortex occurred spontaneously. A single i.p. injection of Pc at doses of 1,000,000, 1,500,000 or 2,000,000 IU/kg resulted in a transient increase in SWD activity. For the latter two doses, Pc injections also induced convulsive activity, i.e. single spikes and trains of spikes accompanied by myoclonies. When 1,500,000 IU/kg of Pc was administered repeatedly (six injections, one every 48 h), the amount of the convulsive activity induced by successive injections decreased but the increase in the number of SWD bursts became more pronounced. This result gives rise to some questions about the development of tolerance to the epileptogenic Pc effects in the course of repeated administration of this antibiotic.

Animals↗
PubMed1994

Oxfendazole treatment of non-parasitized lambs and its effect on the immune system.

Ten parasite-free lambs were drenched with oxfendazole on days 0 and 28 and, one day after each drench, were injected with human erythrocytes and ovalbumin. Ten other antigen-injected lambs were not drenched (controls). Lymphocytes collected 3 days after each antigen injection and cultured in RPMI 1640 plus 5% fetal calf serum (FCS) and lymphocytes collected 3 days after the first and 3 and 7 days after the second antigen injection and cultured in 50% autologous serum had decreased blastogenic activity compared with control lymphocytes. After the second drench, decreased blastogenesis was seen with lymphocytes collected on days 3 and 7 and cultured in 5% FCS and concanavalin A (Con A) and on day 3 when cultured in 5% FCS and phytohaemagglutinin (PHA). Decreased blastogenesis was also seen with lymphocytes collected 7 and 29 days after the second injection of antigen and cultured in 50% autologous serum plus Con A and on days 3, 7 and 29 when cultured in 50% autologous serum and PHA. Significantly depressed antibody responses to both antigens were seen after the second drench. The serum complement level was depressed 3 days after the second injection of antigen. Serum nitric oxide levels were significantly depressed 3 and 21 days after the first and 7 and 21 days after the second injection of antigen. There were no differences in levels of growth-promoting hormones but the drenched lambs gained significantly more weight than the controls.

Animals↗
PubMed1994

A technique for the isolation and purification of viable mucosal mast cells/globule leukocytes from the small intestine of parasitised sheep.

Romney sheep, 1-2 years old, immunized by at least three anthelmintic abbreviated infections of 80-100,000 Trichostrongylus colubriformis larvae usually produced high numbers of intestinal mucosal mast cells/globule leukocytes (MMC/GLs). In isolating these cells, the importance of maintaining the intestine at 37 degrees C, removal of mucus with dithiothreitol, enzymatic dispersion and careful in vitro handling procedures for maximising cell viability are emphasised. The MMC/GLs were separated from most contaminant cells by using a Percoll discontinuous gradient. MMC/GLs collected at the 60/100% Percoll interface were passed through a complement coated nylon wood column to remove the contaminating eosinophils. Viable MMC/GLs were able to grow in vitro in the presence of Concanavalin A and survive in culture for up to 30 days. The MMC/GLs were readily identified by ultraviolet light microscopy after staining with auramine O.

Animals↗
PubMed1994

Sheep small intestinal mast cells/globule leucocytes: ability to divide.

The number of mucosal mast cells/globule leukocytes (MMC/GLs) increase in the intestinal mucosa in response to nematode parasite infections but it is not known if this accumulation is due to in situ cell division, derivation from elsewhere or some combination of both. To determine if MMC/GLs can divide, cells were obtained from immunized Romney sheep and cultured in vitro in RPMI 1640. For cultures supplemented with 10, 20 or 30% foetal lamb serum (FLS) or foetal calf serum (FCS) and without concanavalin A (Con A), cell division had ceased by day 2, but with Con A (3 micrograms ml-1) cell division continued to day 9. Better growth of cells was obtained with the higher concentrations of serum. However the use of 30% or 50% autologous serum with Con A lead to cell death but 10% serum permitted limited growth. The detrimental effect of autologous serum could be overcome by increasing the Con A concentration. It was established that an alpha-macroglobulin present in autologous serum can bind Con A. This macroglobulin appears to have a higher avidity for Con A than does the receptor(s) on the surface of MMC/GLs. Our data seem to indicate that a direct interaction of Con A with the cell surface or a receptor(s) is responsible for MMC/GLs division.

Animals↗