Tissue-type plasminogen activator after venous occlusion in pregnancy and puerperium.

Pregnancy is associated with depressed fibrinolysis as judged from the decreased fibrinolytic response to venous occlusion. In order to elucidate if this decreased response is due to an increase in plasminogen activator inhibitors 1 and 2 (PAI-1, PAI-2), and/or to decreased release of tissue-type plasminogen activator (t-PA) antigen during venous occlusion, 36 women (18 women with normal pregnancy and 18 with gestational hypertension without proteinuria) were followed during pregnancy and puerperium. In each women a 20 min venous occlusion was performed in the second and in the third trimester of pregnancy and 3 days after delivery. The increase in t-PA antigen after venous occlusion relative to basal value was in the second trimester of pregnancy on average 3.7 fold, in the third trimester 4.4 fold, and so not reduced compared to non-pregnant women (3.7 fold increase). After delivery the increase in t-PA antigen was significantly enhanced (8.5 fold, p < 0.005). The fibrinolytic response to venous occlusion measured by euglobulin and t-PA activity was significantly decreased in the third trimester compared to non-pregnant values (both p < 0.005) and returned to somewhat higher (euglobulin clot lysis) or significantly higher (t-PA activity, p < 0.01) values 3 days after delivery. Decreased euglobulin and t-PA activity after venous occlusion in the third trimester coincided with significant increases in basal PAI activity, PAI-1 antigen and PAI-2 antigen (2.9, 2.5 and > 30 fold increase relative to non-pregnant values, respectively, all p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Infection of small mammals with Borrelia burgdorferi sensu lato in Slovenia as determined by polymerase chain reaction (PCR).

Thirty-four small mammals collected in the vicinity of Ljubljana were tested for the presence of Borrelia burgdorferi sensu lato by polymerase chain reaction (PCR) of urinary bladder tissues, using universal flagellin primers and species specific rRNA primers. Seventeen small mammals (50%) were found to be positive, and 7 small mammals were infected with two species of B. burgdorferi sensu lato simultaneously. The most commonly found species was B. afzelii (n = 14), followed by B. burgdorferi sensu stricto (n = 7) and B. garinii (n = 3), as determined by species-specific primers. We conclude that PCR is a rapid and reliable method to detect infection with B. burgdorferi sensu lato in small mammals.

Comparison of PCR methods and culture for the detection of Borrelia spp. in patients with erythema migrans.

The sensitivities of two PCR assays and culture were compared for the detection of Borrelia spp. in skin specimens of 150 patients with typical erythema migrans. In addition, the accuracy of the methods for the identification of Borrelia spp. was compared by analysing culture isolates and material obtained directly from skin biopsy specimens. Borrelia burgdorferi sensu lato was isolated from 73 (49%) of 150 skin biopsy specimens. Using a nested PCR targeting the rrf-rrl region and a PCR targeting the flagellin gene, 107 (71%) and 36 (24%) specimens, respectively, were positive. With both PCRs, positive results were more frequent with culture-positive samples (67/73 (92%) and 24/73 (33%) for the nested and flagellin PCRs, respectively) than with culture-negative samples (40/77 (52%) and 12/77 (16%) for nested and flagellin PCR, respectively). Pulsed-field gel electrophoresis after MluI restriction identified 69/73 (95%) isolates, of which 58/69 (84%) were Borrelia afzelii and 11/69 (16%) were Borrelia garinii. After MseI restriction of PCR products amplified from the intergenic rrf-rrl region, B. afzelii was identified in 73/107 (68%) samples, B. garinii in 22/107 (21%) samples, and both species in 11/107 (10%) samples. The corresponding results for culture-positive specimens were 41/69 (59%), 14/69 (20%), and 7/69 (10%). Comparison of the results for specimens positive according to both approaches revealed complete uniformity in 80% of the cases. Overall, nested PCR was the most sensitive method for the demonstration of Borrelia spp. in erythema migrans skin lesions, followed by culture and PCR targeting the flagellin gene. The congruence of identification results obtained by analyzing culture isolates and material obtained directly from skin biopsies was relatively high but incomplete.

Serological screening for toxoplasmosis in pregnancy in Slovenia.

In the period from 1981 to 1994, serological screening for toxoplasmosis was carried out in 20,953 pregnant women in Slovenia. Seropositivity among pregnant women was found to have decreased from 52% in the 1980s to 37% in the recent period, 1991-94, while during the same period the incidence of suspected primary infections acquired in pregnancy rose from 0.33% to 0.75%. These latest figures ought to promote an informed debate on the possible need for obligatory serological screening of pregnant women in Slovenia for toxoplasmosis.

Isolation of DNA from archival Papanicolaou stained cytological smears using a simple salting-out procedure.

DNA from archival Papanicolaou stained and unstained cytological smears was successfully isolated using a simple, rapid and inexpensive salting-out procedure. The quality of DNA was controlled by polymerase chain reaction (PCR) amplification of segments of the human beta-globin, human beta-actin and human papillomavirus L1 genes. Only negligible differences in amplification efficiency were observed between DNA isolated from stained and unstained smears. The salting-out procedure is a more rapid method for the isolation of DNA than phenol-chloroform extraction and may be used in instances where fresh or cryopreserved clinical specimens are not available.

Incidence of congenital toxoplasmosis in the Republic of Slovenia.

Over a 12-month period, 3959 pregnant women were systematically tested with the Sabin-Feldman dye test in order to assess the incidence of congenital toxoplasmosis in Slovenia. The results suggest that this is approximately 3/1000 live births. This relatively high incidence of congenital toxoplasmosis in Slovenia may make the costing of preventive screening programmes justifiable.