Recurrent antiphospholipid-related deep vein thrombosis as presenting manifestation of systemic lupus erythematosus

Antiphospholipid antibodies (aPL) are frequently associated with thrombotic disorders in the so-called antiphospholipid syndrome. Together with anticardiolipin antibodies (aCL), lupus anticoagulant (LA) is the main diagnostic tool for aPL detection. Since LA determination is based on the finding of prolonged clotting time in vitro, concomitant anticoagulant therapy may significantly interfere with its detection. We report a case of a boy in whom recurrent aPL-related thrombosis heralded for several months the onset of systemic lupus erythematosus (SLE). Abnormally increased in vitro clotting times at the time of the second thrombotic event led to the suspicion of the presence of LA activity. However, this latter finding was difficult to interpret since the patient was already on heparin treatment at the time of our first observation. Thus, LA was assayed using a commercial kit in which a heparin neutralizer is included (Staclot LA). Two consecutive samples from the patient were compared with eight patients on anticoagulant therapy for non-aPL-related thrombotic events and 20 healthy controls. The study showed that, taking into account the concomitant anticoagulant treatment, Staclot LA was positive only in the propositus, raising the suspicion of a possible aPL-related origin of the thrombotic event. This issue was definitively confirmed in a subsequent follow-up. Conclusion The present report shows that aPL-related deep vein thrombosis can be the earliest clinical manifestation of pediatric SLE, and that Staclot LA may have a role in LA detection during the course of anticoagulant treatment. Received: 6 October 1998 / Accepted: 15 September 1999     

Alcohol reduces MMP-2 in humans and isolated smooth muscle cells

Alcoholic beverages are known to exert a protective effect on atherosclerotic disease. This study aimed to assess the in vivo and in vitro effects of alcohol on matrix metalloproteinase (MMP) -2 and -9, known to determine atherosclerosis progression. Eighteen healthy volunteers, regular drinkers (two standard alcohol servings/day, on average) at first examination (baseline) were asked to abstain from any alcoholic beverage for one week (abstention), and then to assume two standard alcohol servings of beer daily for 1 week (re-exposure). Activity of MMP-2 and -9, total antioxidant activity (AOA), glutathione (GSH) plasma levels were carried out at baseline, at the end of abstention, and after 1 week of re-exposure. To validate the in vivo results, MMP-2 activity and expression, AOA, and GSH, were determined in human smooth muscle cells treated for 96 h with increasing concentrations (12.5-100 mM) of ethanol. MMP-2, but not MMP-9 plasma activity was higher at abstention than at baseline or re-exposure (P<.001 and P< or =.005, respectively). Changes in AOA and GSH throughout the study were not significant. No correlation was found between MMPs and antioxidant activity. In vitro, ethanol at 25 mM reduced by around 10% MMP-2 activity (P=.003) in smooth muscle cells, whereas MMP-2 expression, AOA, and GSH were unaffected. Alcohol reduces MMP-2 plasma activity in healthy humans and in isolated vascular smooth muscle cells. This in vitro reduction is unrelated to MMP-2 expression in vascular cells or to antioxidant levels changes.     

Expression of HER-2/neu oncoprotein, DNA-ploidy and S-phase fraction in advanced ovarian cancer

The immunohistochemical expression of HER-2/neu and cytofluorimetric data were retrospectively analyzed in a group of primary advanced ovarian cancers. Thirty-three out of 94 (35%) cases showed a specific p185/neu immunoreaction. No correlation between p185/neu expression and any of the clinico-pathologic parameters examined was observed. As far as cytofluorimetric data are concerned, 38 out of 69 (55%) of the tumors were diploid (DNA index = 1) while 31 (45%) were aneuploid (DNA index from 1.10 to 2.50 with a median value of 1.50). Ovarian tumors were defined as of low and high S-phase fraction in 68% and 32% of the cases, respectively. Tumor ploidy and S-phase fraction did not correlate with the clinico-pathologic characteristics or p185/neu oncoprotein expression. Aneuploid tumors had a higher S-phase fraction (mean: 15.81 ± 13.44) than diploid tumors (mean: 8.89 ± 7.98) ( P < 0.01). p185/neu expression failed to affect significantly both overall and progression free survival. On the other hand tumor ploidy was found to be related to the prognosis of advanced ovarian cancer patients although the difference was not statistically significant. As far as progression free survival is concerned, the median time to recurrence was not reached for diploid cases whereas it was 21 months for aneuploid cases ( P < 0.05). The 5-year survival for patients with a low S-phase fraction (58%) was significantly higher than for patients with high S-phase fraction tumors (28%) ( P < 0.01). Median time to recurrence was 48 and 17 months for low and high S-phase fraction tumor patients, respectively ( P < 0.05). However, in a multivariate analysis both tumor ploidy and S-phase fraction did not retain their prognostic value. The assessment of the role of the parameters examined in improving the prognostic characterization of ovarian cancer patients should be investigated in large multicenter clinical trials.     

7-Alkyldeoxyguanosine adduct detection by two-step HPLC and the 32P-postlabeling assay

7-Alkyldeoxyguanosine DNA adducts may be a marker for some N-nitrosocompound exposures and subsequent human cancer risk. A sensitiveand highly specific assay for the detection of 7-methyl-2'-deoxyguanosine-3'-monophosphate(7-methyldGp) and 7-ethyl-2'-deoxyguanosine-3'-monophosphate(7-ethyldGp) has been developed by combining two different HPLCpurification steps with the ³²P-postlabeling assay. We previouslyreported that ion-pair reverse-phase (IP) chromatography coupledwith the ³²P-postlabeling assay detects 7-methyldGp in humanlung, but have found that other nucleotides and unknown adductsco-elute. Thus, weak anion exchange (AE) HPLC was added in tandemwith IP HPLC prior to the ³²P-postlabeling assay. 2'-Deoxyguanosine-3'-monophosphate(dGp) is incorporated into the assay as an internal standardfor the assessment of enzyme labeling efficiency and adductrecovery. The methodology was validated using radiolabeled DNAand liquid scintillation counting, which accounts for adductloss from enzymatic digestion to detection. Levels of 7-ethyldGpalso were correlated with accelerator mass spectrometry. Theoverall adduct recovery with this method was 58% for 7-methyldGpand 98% for 7-ethyldGp. The detection limit for both assaysusing 100 µg of DNA was one adduct in 10⁸ unmodified dGp.7-MethyldGp and 7-ethyldGp levels were determined in ten humanlung samples at levels of 1.4–5.4 and 0.6–3.1 adductsper 10⁷ dGp respectively, and in five human lymphocyte samplesat levels of 5.0–8.3 and 0.3–1.4 adducts per 10⁷dGp respectively. Combining the two HPLC purification stepsand the ³²P-postlabeling assay attains chemical specificity,retains sufficient quantitative sensitivity and should be usefulin human biomonitoring studies.     

Enterovirus surveillance of Italian healthy children

Surveillance of acute flaccid paralysis (AFP) is the golden strategy recommended by the WHO to verify the condition of polio eradication in a country. Because of the difficulty to detect all of the expected AFP cases and to reach the target incidence of 1/100,000 requested by WHO, the surveillance of enteroviruses in the population has been adopted by several countries as an important additional method to verify the absence of wild-poliovirus circulation. To complete the results of AFP surveillance set up in Italy in 1996, we have conducted a wild poliovirus surveillance by examining stool samples from 1551 healthy children aged less than 5 years, collected during the period January 1997 to January 1998. The children were from three cities (Parma, Rome and Bari) located in northern, central and southern Italy. Thirty-nine polioviruses, 72 non-polio enteroviruses and 50 enteric, non-entero (NE), viruses were isolated from stool specimens. Polioviruses identified were nine type 1, seven type 2 and twenty-three type 3. Characterization of isolates by both antigenic and molecular methods showed that all polioviruses were of vaccine origin. As expected, most polioviruses, especially types 2 and 3, presented retromutations known to be associated with loss of the Sabin attenuated phenotype. The results of this study support the data obtained from the active AFP surveillance conducted in Italy in the same period – on the absence of paralytic disease due to wild poliovirus – and altogether demonstrate the effectiveness of the vaccination program.     

The screening of tubal abnormalities in the infertile couple

Purpose: Our goal was to assess, with a prospective study, the role of hysterosalpingo-contrast sonography (HyCoSy) with an echocontrast agent and transvaginal ultrasonography alone in the evaluation of tubal status. Methods: Thirty patients were included in the study. These patients underwent an initial plain transvaginal ultrasound examination the day before the HyCoSy. The findings obtained from both examinations were compared with laparoscopic diagnosis, performed in the same menstrual cycle. Results: The kappa values were 0.48 for patency evaluation and 0.67 for the diagnosis of the presence of at least one patent tube, suggesting a good agreement in both cases between HyCoSy and surgery. HyCoSy had a significantly lower sensitivity (50%), but not a significantly higher specificity (75%), than transvaginal ultrasonography alone in the diagnosis of tubal infertility-related abnormalities such as peritubal adhesions. Conclusions: The study demonstrates that the HyCoSy is a useful test when scheduling the most suitable treatment for infertile couples.     

The screening of embryonic viability in early asymptomatic pregnancy by a single endosonographic scan associated with plasma human chorionic gonadotropin determination

Purpose Our purpose was to assess, with a prospective study with random assignment of the day of the first evaluation, whether a single transvaginal ultrasonographic evaluation together with the determination of plasma hCG levels could be used to screen embryonic viability in early asymptomatic pregnancy.Methods In 260 pregnant women observed from January 1991 to November 1993 with spontaneous pregnancies where the exact date of ovulation was known, a single transvaginal ultrasonographic measurement of gestational sac with determination of plasma hCG levels, transformed to their natural logarithm (lnhCG), was performed. An abnormal result was defined as a value of lnhCG per mean gestational sac below the 95% lower confidence limit of the viable pregnancy group.Results The sensitivity was 31%, with a specificity of 97%.Conclusion The study demonstrates that this method has a poor predictive capacity to distinguish viable pregnancy from nonviable pregnancy with a kappa value less than 0.4.