Intraspecific variation between the ITS sequences of Toxocara canis, Toxocara cati and Toxascaris leonina from different host species in south-western Poland.

Some parasitic nematodes can inhabit different definitive hosts, which raises the question of the intraspecific variability of the nematode genotype affecting their preferences to choose particular species as hosts. Additionally, the issue of a possible intraspecific DNA microheterogeneity in specimens from different parts of the world seems to be interesting, especially from the evolutionary point of view. The problem was analysed in three related species - Toxocara canis, Toxocara cati and Toxascaris leonina - specimens originating from Central Europe (Poland). Using specific primers for species identification, internal transcribed spacer (ITS)-1 and ITS-2 regions were amplified and then sequenced. The sequences obtained were compared with sequences previously described for specimens originating from other geographical locations. No differences in nucleotide sequences were established in T. canis isolated from two different hosts (dogs and foxes). A comparison of ITS sequences of T. canis from Poland with sequences deposited in GenBank showed that the scope of intraspecific variability of the species did not exceed 0.4%, while in T. cati the differences did not exceed 2%. Significant differences were found in T. leonina, where ITS-1 differed by 3% and ITS-2 by as much as 7.4% in specimens collected from foxes in Poland and dogs in Australia. Such scope of differences in the nucleotide sequence seems to exceed the intraspecific variation of the species.

Developmental age, physical fitness and Toxocara seroprevalence amongst lower-secondary students living in rural areas contaminated with Toxocara eggs.

Scarce and inconclusive information on general biological impact of Toxocara invasion on paratenic hosts, and people in particular, has led us to undertake a comprehensive study of the problem. The study has been conducted in a rural environment, which is considered a toxocarosis risk factor. In total 200 soil samples have been screened for Toxocara eggs by flotation, of which 14.5% were positive. Backyards close to households were most heavily contaminated with infectious eggs--21.7% of positive samples. ELISA serological tests performed on 242 lower-secondary students found 14.5% of the studied population to be definitely positive--16.5% of boys and 12.8% of girls, respectively. The odds of being infected with Toxocara were 2 times (CI: 1.15-3.85) more likely for individuals who owned a cat than those who did not own a cat. Strong significant correlation between seropositivity and the presence of a dog in a household was found with boys. The level of developmental age was significantly higher in seropositive than in seronegative students. No significant correlation has been observed between the motor abilities and seropositivity of students. Seropositive boys had significantly lower end-of-year grades than their seronegative counterparts.

Utilizing a polymerase chain reaction method for the detection of Toxocara canis and T. cati eggs in soil.

A polymerase chain reaction (PCR) technique has been used for the differentiation of T. canis and T. cati eggs isolated from soil and previously identified from microscopical observations. The method, using specific primers for the identification of the two Toxocara species, was assessed in both the field and laboratory. Successful results were obtained when only a single or large numbers of eggs were recovered from 40 g soil samples. The method is sensitive, allows analysis of material independent of the stage of egg development and can be adapted for the recovery of other species of parasites from soil.

Adult croup--a case report.

A case of a female adult patient with croup is described. Inflammation, as well as pseudomembranes, restricting the patency of trachea, developed within several hours after the first symptoms of infection were observed. Tracheostomy was performed Bronchofiberoscopies were repeated on a regular basis several times a day over a period of two weeks, with the removal of fibrinous casts and dense secretion being the only way to save the patient's life.

[Soil contamination with Toxocara spp. eggs in the Elblag area]. / Zanieczyszczenie gleby jajami Toxocara spp. na terenie elblaga.

The distribution of Toxocara spp. eggs in Elbl4g was studied. Out of 72 soil samples collected in public places of the city 13.9% were positive and the mean egg density was 3.75/100g soil. The city backyards were much more contaminated with Toxocara spp. eggs (18.0%) than the playgrounds (4.5%). In sandpits examined the eggs were not found. Almost 80% of Toxocara spp. eggs recovered were infective. T. cati eggs were more frequent than T. canis eggs. Additionally in examined samples two eggs of Ancylostoma caninum and one egg of Ascaris lumbricoides were recognized.

[Distribution of sources of Toxocara spp. infection in urban and rural environments in Poland]. / Rozmieszczenie zródel inwazji Toxocara spp. w srodowisku miejskim i wiejskim w Polsce.

Soil examinations made in 5 regions of Poland in the 1990s comprised 1184 samples taken from urban areas and 590 samples from suburban and rural areas. Toxocara spp. eggs were found more often in urban areas (14% positive samples) than in suburban and rural once (12% positive samples). The average ratio of positive samples was: in the streets and roads--19.3% (0.36 eggs/100g of soil), near houses (backyards, gardens)--18.6% (1.11 eggs/100g), in sandpits--13.0% (0.23/100g), in parks and public gardens--10.5% (0.46 eggs/100g), on playgrounds and playing fields 9,4% (0,06 eggs/100g) and on the beaches--3.4% (0.03 eggs/100g). In children habitual play areas the prevalence of T. cati eggs was higher than T. canis eggs.

[Proangiogenic cytokines (bFGF and VEGF) in BALF from two different lung segments examined by high resolution computed tomography (HRCT) in patients with sarcoidosis]. / Cytokiny proangiogenne (bFGF i VEGF) w BAL z dwóch róznych segmentów pluc wskazanych badaniem tomografii komputerowej o wysokiej rozdzielczosci (HRCT) u chorych na sarkoidoze.

The aim of the study was to evaluate the concentrations of bFGF and VEGF in double BAL (2 x 120 ml) from two different lung segments: (s.A) from upper lobe with the most and (s.B) from lower lobe with the least extensive involvement estimated by high resolution computed tomography (HRCT). Examined group consisted of 28 sarcoid patients with homogeneous, regular distribution of nodular opacities in conventional chest X-ray (14 F, 14 M aged 19-54). Eleven healthy volunteers served as controls. In patients with sarcoidosis we observed the significantly higher levels (p < 0.01) of bFGF (1.79 pg/ml, 1.48 pg/ml) and VEGF (107.5 pg/ml, 109.7 pg/ml) in BAL from s.A and s.B respectively in comparison with BAL from lung segments Abis and Bbis in control group (bFGF: 0.75 pg/ml, 0.47 pg/ml and VEGF: 33.7 pg/ml, 43.9 pg/ml respectively). bFGF in BAL from s.A in active sarcoidosis was higher than in s.A and s.B in non-active sarcoidosis. Concentrations of bFGF in BAL from both s.A and s.B correlated positively with CD4/CD8 ratio and absolute number of lymphocytes, CD4 cells and lymphocytes HLA-DR estimated in BAL from these lung segments. We conclude that bFGF and VEGF may be involved in sarcoidosis pathogenesis and bFGF may be useful in estimation of sarcoidosis activity.

[Cytokines TNF-alpha and GM-CSF in BAL from two different segments of lungs defined by high resolution computerized tomography (HRCT) in patients with sarcoidosis]. / Cytokiny TNF-alpha i GM-CSF w BAL z dwóch róznych segmentów pluc wskazanych badaniem tomografii komputerowej o wysokiej rozdzielczosci (HRCT) u chorych na sarkoidoze.

The aim of the study was to evaluate the concentrations of TNF-alpha and GM-CSF in double BAL (2 x 120 ml) from two different lung segments: (s.A) from upper lobe with the most and (s.B) from lower lobe with the least extensive involvement estimated by high resolution computed tomography (HRCT). Examined group consisted of 28 non-smoking sarcoid patients with homogenous, regular distribution of nodular opacities in conventional chest X-ray (14 F, 14M aged 19-54). In examined patients 16 had nonhomogenous distribution (ND) and 12 had regular distribution (RD) of HRCT changes. Eleven healthy volunteers served as controls. In patients with sarcoidosis we observed the significantly higher concentrations (p < 0.01) of TNF-alpha (3.18 pg/ml, 2.64 pg/ml) and GM-CSF (1.01 pg/ml, 0.95 pg/ml) respectively in BAL fluid from s.A and s.B in comparison with BAL from s.Abis and s.Bbis in control group (TNF-alpha: 0.46 pg/ml, 0.47 pg/ml and GM-CSF: 0.28 pg/ml, 0.31 pg/ml respectively). Mean concentration of TNF-alpha in BAL from s.A (3.77 pg/ml) in ND group was significantly higher than in BAL from s.B in RD group (2.91 pg/ml). TNF-alpha in BAL from s.A in active sarcoidosis was higher than in BAL from s.A and s.B in non-active sarcoidosis. Concentrations of TNF-alpha in BAL from both s.A and s.B correlated positively with CD4/CD8 ratio, percentage of lymphocytes, lymphocytes HLA-DR+ and absolute number of CD4 cells and negatively with CD8 cells estimated in BAL from these lung segments. In patients with indications to therapy the level of GM-CSF in BAL from s.A (1.44 pg/ml) was significantly higher (p < 0.05) than in BAL from s.A (0.64 pg/ml) in patients without indications to treatment. We conclude that TNF-alpha and GM-CSF may be involved in sarcoidosis pathogenesis and TNF-alpha may be useful in estimation of sarcoidosis activity.

Effect of proline rich polypeptide from ovine colostrum on virus replication in human placenta and amniotic membrane at term; possible role of endogenous tumour necrosis factor alpha.

Freshly prepared organ cultures of human placentae and amniotic membranes at term show different sensitivity to vesicular stomatitis virus (VSV) infection. In six of 16 amniotic membranes and seven of 17 placentae VSV replicated to relatively high titres (10(3)-10(6)TCID(50)/ml). The others were partially or completely resistant to virus infection (<10(1)-10(2)TCID(50)/ml). Addition of the immunomodulating agent, proline-rich-polypeptide (PRP) from ovine colostrum to explants freshly obtained from the organs, influenced VSV replication in a manner dependent on the innate immune state of the organ culture. In cultures resistant to the virus, PRP at a concentration of 10 microg/ml increased 10-10 000 times the VSV titre. In contrast, treatment of highly sensitive cultures by PRP hardly influenced viral replication at all. The effect of virus stimulation by PRP was abolished by specific anti-TNF antibodies. The results indicate that endogenous TNF may be one of the mediators of virus stimulation by PRP. Antibodies against TNFalpha, added to VSV infected organ cultures sensitive to the virus reduced viral replication. The antibodies caused stimulation of virus replication in VSV-infected resistant organ cultures. The results indicate the double role of endogenous TNF in viral replication in placenta and the amniotic membrane.

Effect of viral infections on the ability of human endothelium for interferon, tumor necrosis factor and interleukin 6 production.

We have reported that cultured human umbilical cord vein endothelial cells (HUVEC) differ from endothelium present on vein surface of organ culture (OC) in production of cytokines and susceptibility to viral infections. In this paper we present the effect of viral infections on interferon (IFN), tumor necrosis factor (TNF), and interleukin 6 (IL-6) production in two culture systems: HUVEC and OC. Infection of 24-48 h HUVEC with herpes simplex type 1 (HSV-1) and vesicular stomatitis virus (VSV) reduced the amounts of IL-6 and TNF produced in comparison to those released spontaneously by uninfected cells. No IFN was detected in media from infected and uninfected HUVEC. Limited viral infections of 3-h-HUVEC and OC usually diminished their efficiency of IL-6 and TNF production. In the case of IL-6 synthesis by OC, effect of viral infection depended, however, on the constitutive synthesis of the cytokine. When spontaneous production was high (> 800 U/ml), VSV and HSV-1 infection reduced IL-6 level by 2-50 times; in the case of low production (< 150 U/ml) the stimulation effect (2-4 fold) was observed. OC released spontaneously some IFN activity (2-32 U/ml). HSV-1 infection of OC reduced IFN level, while VSV in single cases slightly upregulated IFN synthesis.

Antiviral nonspecific immunity of human placenta at term: possible role of endogenous tumor necrosis factors and interferons.

The antiviral immunity of human placenta and amniotic membrane in an organ culture (OC) system was studied. Freshly isolated explants of most of the placentas at term and the amniotic membranes were found to be relatively resistant to herpes simplex virus type 1 (HSV-1), encephalomyocarditis virus (EMCV), and vesicular stomatitis virus (VSV) infections. After in vitro aging, however, the OC acquired the sensitivity to the viruses. In about 66%-90% of placentas, resistance of freshly isolated explants to the infection was observed. This indicates that the placentas displayed a constitutive immunity against the viruses. To study the role of endogenous cytokines in antiviral immunity, we added specific antibodies neutralizing IFN and TNF activities to VSV-infected OC and checked their influence on viral replication. Increases of 10-fold to 100-fold of VSV replication in the OC treated with anti-TNF-alpha, anti-IFN-alpha, anti-IFN-gamma or anti-IFN-beta sera were observed. The results indicate the importance of the endogenous cytokines in placental and amniotic membrane immunity. However, we did not observe a simple correlation between the spontaneous IFN and TNF production and the level of resistance against viruses. In view of the results, the participation of TNF and IFN in the constitutively expressed immunity of human placenta is of a more complex nature.

Thallium-201 myocardial perfusion imaging in patients before and after successful percutaneous transluminal coronary angioplasty.

We studied the value of exercise thallium-201 (Tl-201) scintigraphy for evaluation of myocardial perfusion improvement and the detection of restenosis in patients after successful percutaneous transluminal coronary angioplasty (PTCA). Fifty-three patients (43 male and 10 female) ages 38-71 years (mean 55.3) were analysed. Exercise Tl-201 scintigraphy was performed before PTCA, and 6-10 days and then 3-6 months after the procedure. In all patients repeated coronary angiography was done 3-6 months after PTCA. Before PTCA myocardial perfusion defects were observed in all patients. Immediately after PTCA, an improvement in myocardial perfusion was noted in 36 patients (61%). Total normalisation of the scintigraphic picture was observed in only 12 patients. Coronary angiography after 3-6 months showed patency of dilated vessels in 11 out of those 12 patients (91.3%). In scintigraphy, performed 3-6 months after PTCA, a normal scan was present in 20 patients and recurrence of stenosis was found in only 2 of those 20. Stenosis was found in 22 (60%) of 33 patients with perfusion defects. For the purpose of describing the character of the myocardial perfusion changes, statistical analysis of a number of segments was performed. The predictive value of Tl-201 scintigraphy for detection of restenosis was established. The positive value for the procedure performed 6-10 days after PTCA was 56%, and the negative value of prediction of restenosis was 91%. Three to 6 months after PTCA, a high negative value of scintigraphy was observed (-90%) and a low positive predictive value was still present (63%).

[Scintigraphic evaluation of myocardial perfusion after using TI-201 in patients before and after successful percutaneous transluminal coronary angioplasty]. / Scyntygraficzna ocena perfuzji miesnia sercowego przy uzyciu talu 210 u chorych przed i po skutecznej angioplastyce naczyn wiencowych.

We analysed 53 patients (43 men and 19 women) age 38-71. Exercise TI-201 scintigraphy was performed before PTCA, 6 to 10 days and 3 to 6 months after PTCA. Before PTCA in all the patients myocardial perfusion detects on scintigraphy were observed. The imaging performed 6 to 10 days after PTCA showed an improvement seen as a decrease in the number of ischaemic segment in 36 patients (67.9%) and total normalisation of scintigraphic picture in 12 patients. Coronary angiography performed 6 months after PTCA showed patency of the dilated vessel in 11 (91.3%) among these patients. In exercise TI-201 scintigraphy performed 3 to 6 months after PTCA normalised scan was observed in 20 patients, recurrence of stenosis was found only in 2 (10%) of those patients. In 33 patients with transient perfusion defects, angiographic restenosis was found in 22 (60%) patients. Predictive value of exercise TI-201 scintigraphy for occurrence of restenosis was established. Positive predictive value of the study performed 6 to 10 days after PTCA was 56%. Negative predictive values of such a study was 91%. Similarly, for detection of restenosis in scan performed 3 to 6 months after PTCA there was found a strong negative predictive value-90% and a weak positive predictive value-63%.