Chlamydophila pneumoniae and human cytomegalovirus in atherosclerotic carotid plaques--combined presence and possible interactions.

The aim of our study was to investigate the combination of Chlamydophila pneumoniae and human cytomegalovirus (HCMV) as a pathogenic factor in atherosclerosis. Accordingly, we tested by means of PCR and immunohistochemistry the presence of these pathogens in the same atherosclerotic carotid specimen. The histology of the samples and the patients' antibodies against these pathogens were evaluated. Further, we examined the impact of C. pneumoniae and HCMV infection on the gene expression of the human monocytic cell line U937. Six of the 22 samples contained only C. pneumoniae, 4 contained only HCMV, 7 contained both C. pneumoniae DNA and/or antigens of both pathogens, and 5 samples were negative. No correlation was found between the presence of these microbes and either the cellular structure of the plaques, or the serostatus of the patients. The infection of U937 cells with HCMV and especially C. pneumoniae induced inflammation and atherosclerosis-related genes. Furthermore, the doubly-infected cells produced higher levels of the mRNA of pro-platelet basic protein and fatty acid binding protein 4. In conclusion, C. pneumoniae is often present in combination with HCMV in atherosclerotic carotid lesions. The in vitro coinfection model reveals that the doubly-infected monocytes are potent expressors of proatherosclerotic genes, suggesting that this coinfected population may accelerate the process of atherosclerosis.

Repeated biopsies in evaluation of therapeutic effects in prostate carcinoma.

BACKGROUND: Apoptosis is one of the major events following total androgen blockade (TAB). The aim of this study was to determine the predictive value of some histological parameters including apoptosis and gene products which influence apoptosis, based on repeated biopsies taken from the same patients. METHODS: At the time of diagnosis by needle biopsy TNM stage, serum PSA, Gleason's grade, apoptotic and mitotic index, Ki67, p53, and bcl(2) expression were investigated in 60 prostate carcinoma patients. Antiandrogen therapy supplemented with surgical or chemical castration was administered. Serum PSA-test and needle biopsy were repeated 13-14 weeks after starting the therapy, simultaneously with determination of the apoptotic and mitotic index, Ki67, p53, and bcl(2) expression. RESULTS: Forty-seven patients were alive at the end of the study, 13 patients died. Decrease in mitotic, increase in apoptotic index predicted favourable long-term response to antiandrogen therapy. Lower Ki67 and (mutant) p53 expression in the first and also in the second biopsy pointed to favourable effect of antiandrogen treatment. Since the ratio between Ki67 and apoptotic index strongly decreased in the survivors upon therapy, changes in Ki67/apoptosis ratio is recommended as a histologically detectable predictive factor. bcl(2) expression did not show significant correlation with the outcome of the disease. CONCLUSIONS: Histological evaluation of mitotic and apoptotic index, Ki67, and p53 expression in repeated biopsies contributes to predicting the value of the actual treatment and may be useful to institute alterations in therapy.

[The role of uterine and umbilical artery Doppler velocimetry for the early detection of pathologic pregnancy]. / Az arteria uterina és az arteria umbilicalis Doppler-ultrahangvizsgálatának szerepe a kóros terhesség korai felismerésében.

Pregnancy induced hypertension remains the most frequent reason of maternal and fetal complications. Recent studies of the entire pathophysiology of the abnormal implantation proved association between the impaired placental performance and the reduced uteroplacental circulation. Therefore the Doppler examination of uterine and umbilical blood flow may provide the early recognition of high-risk pregnancies complicated by maternal hypertension and fetal intrauterine growth retardation. The authors endeavor to describe the details and characteristics of uteroplacental haemodynamic changes during the course of physiologic and defective implantation. On the basis of the currently available scientific publications they summarize the role of the uterine and umbilical artery Doppler velocimetry as a screening tool in the detection of the consequences of the impaired uteroplacental perfusion. They conclude that abnormal velocity waveforms obtained from the uterine and umbilical arteries may help to improve the efficacy of the prenatal care and the perinatal outcome as well. The clinical benefits of this screening method will be precisely defined on the basis of further multicenter studies in Hungary.

Connexin 43 expression in rat aortic smooth muscle after ovariectomy and hormonal replacement.

The purpose of this study was to determine the effects of ovariectomy and long-term combined sexual hormone replacement on the gap junctional protein, connexin 43 (Cx43) of aortic medial smooth muscle cells in rats. Twenty non-pregnant mature Wistar female rats were divided into five groups (four animals in each group). Group A underwent ovariectomy, Group B underwent ovariectomy and received estradiol propionate, Group C underwent ovariectomy and received medroxyprogesterone acetate and Group D underwent ovariectomy and received both hormones. Group E was sham-operated and used as control. After 15 weeks of treatment, thoracic aortas were removed and immunohistochemistry was carried out using a specific fluorescent antibody against Cx43. Tissue sections were examined by confocal laser scanning microscopy and analysed by the Scion Image program. All five different groups had the same distribution and extent of Cx43 in the aorta. Neither the ovariectomy nor the hormone replacement had any effect on the Cx43 expression of aortic smooth muscle cells in rats as compared to control animals. These results indicate that sexual steroids do not influence the gap junctional protein Cx43 of the medial layer of aorta in rats. They may suggest that the beneficial effects of estrogen are not mediated via gap junctions in the human aorta either.

Study of inflammatory responses to crocidolite and basalt wool in the rat lung.

The subacute effects of crocidolite and basalt wool dusts were studied by nmeans of biochemical, morphological. and histological methods 1 and .3 mo after intrabronchial instillation. The cell count, protein and phospholipid contents, and lactate dehydrogenase (LDH) activity were determined in the bronchoalveolar lavage (BAL). Both types of fibers induced a prolonged inflammatory reaction in the lung. All the parameters studied in the experimental groups were more markedly elevated after 3 mo. Relative to the control, the protein and LDH values were increased three- to fivefold, the phospholipid content twofold, and the number of free cells in the BAL exceeded the control level up to ninefold. The inflammatory responses to crocidolite and basalt wool in the lung did not differ significantly. In spite of this, basalt wool is recoinmended as an asbestos substitute, as the use of this man-nade fiber may result in a significantly lower release of dust than that from crocidolite.

Scanning electron microscopical investigations of broncho-alveolar casts after intratracheal asbestos fibre instillation.

The evaluation of the toxicity of mineral fibres has been tried to achieve in experimental animal models. However, the appearance of fibres in the pleural space could not be explained satisfactorily. Histomorphological examinations showed that intratracheal instillation of asbestos fibres leads to parabronchial and intraalveolar granulomatous tissue reactions and bronchial epithelial regenerations. For further elucidation of the pathogenesis of lung cancer and of mesothelioma the localisation and transport of inhaled fibres is of high interest. Thus, a three dimensional visualization of the structure of rat lungs before and after intratracheal instillation of UICC crocidolite fibres was performed by plastic casts to follow the way of asbestos fibres in the lung tissues and the pleura. The casts allowed to demonstrate airway structures with imprints of epithelial cells and blood vessels of normal and treated animals by scanning electron microscopy. Instilled asbestos fibres transformed bronchial structures and resulted in cystic deformations of the pleural surface. The penetration of single fibres through bronchial trunks and the visceral pleura could be shown for the first time in a three-dimensional topography of the affected tissue. Now, there is support for similar results of histomorphological examinations indicating the possibility that asbestos fibres could penetrate the pleura and migrate into the pleural space. The question if the migration of fibres is a mechanical movement or an active transport is still under discussion.

Improving diagnostic accuracy of prostate carcinoma by systematic random map-biopsy.

Systematic random rectal ultrasound directed map-biopsy of the prostate was performed in 77 RDE (rectal digital examination) positive and 25 RDE negative cases, if applicable. Hypoechoic areas were found in 30% of RDE positive and in 16% of RDE negative cases. The score for carcinoma in the hypoechoic areas was 6.5% in RDE positive and 0% in RDE negative cases, whereas systematic map biopsy detected 62% carcinomas in RDE positive, and 16% carcinomas in RDE negative patients. The probability of positive diagnosis of prostate carcinoma increased in parallel with the number of biopsy samples/case. The importance of systematic map biopsy is emphasized.

Are there systemic changes in the arterial biomechanics of intracranial aneurysm patients?

Current theories on the development of intracranial aneurysm suggest that there is a general weakness of vascular connective tissue. Potential systemic alterations in arterial wall biomechanics were tested in the present study. A three-dimensional in vitro stress-strain analysis was made in the 0-200-0 mmHg pressure range on cylindrical segments excised from the anterior cerebral artery, the radial artery and from the arteria dorsalis pedis of aneurysm patients and of control cadavers. In the anterior cerebral artery from aneurysm patients (intracranial artery segments directly not affected by the aneurysm or by the subarachnoid bleeding), we found the wall thickness to be larger (0.1480+/-.019 versus 0.091+/-0.004 mm), the radius/wall thickness ratio smaller (9.7+/-1.4 versus 14.1+/-1.2), and the tangential wall stress lower [(0.122+/-0.019)x10(6) versus (0.181+/-0.016)x10(6) N/m2 at 100 mmHg] than in control subjects. Reduced radius was found in the extremity arteries studied. Elastic parameters, as incremental distensibility and elastic modulus, were remarkable similar. Our study demonstrates changes in the geometry of walls of arteries not directly affected by aneurysm formation, and it thus confirms systemic vascular pathology in this disease. At the same time, these data show that the molecular and morphological defects of arterial connective tissue formation generally thought to induce intracranial aneurysms will probably not affect the components responsible for the passive elastic properties of the vascular wall.

Perinatal outcome in grand and great-grand multiparity: effects of parity on obstetric risk factors.

OBJECTIVE: We sought to compare obstetric and neonatal complications among great-grand multiparous, grand multiparous, and multiparous women. STUDY DESIGN: One hundred thirty-three great-grand multiparas, 314 grand multiparas, and 2195 multiparas who were delivered of their infants between 1988 and 1998 were selected for the study. To facilitate comparison, the patients were all >35 years old and had similar socioeconomic characteristics. RESULTS: The incidence of malpresentation at the time of delivery, maternal obesity, anemia, preterm delivery, and meconium-stained amniotic fluid increased with higher parity, whereas the rate of excessive weight gain and cesarean delivery decreased. Compared with grand multiparas, great-grand multiparas had significantly elevated risks for abnormal amounts of amniotic fluid, abruptio placentae, neonatal tachypnea, and malformations but lower rates of placenta previa (P <.05). The incidence of postpartum hemorrhage, preeclampsia, placenta previa, macrosomia, postdate pregnancy, and low Apgar scores was significantly higher in grand multiparas than in multiparas, whereas the proportion of induction, forceps delivery, and total labor complications was significantly lower than in the multiparous group (P <.05). Similar frequency of maternal diabetes, infection, uterine wall scar rupture, variations in fetal heart rate, fetal death, and neonatal mortality was found in the 3 groups. CONCLUSION: Both high-parity groups have their own risk factors, but the rate of some complications decreases with higher parity. In addition, perinatal mortality remains low in these patients, and therefore, under satisfactory socioeconomic and health care conditions, high parity should not be considered dangerous.

Diagnosing amnionicity at 6 weeks of pregnancy with transvaginal three-dimensional ultrasonography: case report.

OBJECTIVE: To report a case of monochorionic, diamniotic twin pregnancy diagnosed at 6 weeks of pregnancy with three-dimensional (3D) transvaginal sonography and to discuss the use of this diagnostic method in the evaluation of multifetal pregnancy in the first trimester. DESIGN: Case report. SETTING: University-based IVF program. PATIENT(S): A 30-year-old ovum recipient underwent ultrasonographic evaluation of a first-trimester twin pregnancy. INTERVENTION(S): Two-dimensional (2D) and 3D transvaginal sonography. MAIN OUTCOME MEASURE(S): Accurate diagnosis of chorionicity and amnionicity. RESULT(S): Monoamniotic pregnancy and conjoined twinning could not be ruled out by using 2D transvaginal sonography at 6 weeks, because only one yolk sac (YS) and no membranes could be visualized, and the two embryos were closely positioned within one gestational sac. Applying 3D technique, two YSs and two separate embryos could clearly be observed, establishing the correct diagnosis of a monochorionic, diamniotic pregnancy. CONCLUSION(S): The 3D transvaginal ultrasonography provides a quick and accurate diagnostic modality for the evaluation of a first-trimester multiple gestation.

Tenascin expression and angiogenesis in breast cancers.

The expression and the distribution of tenascin as well as the extent of blood vessel formation (angiogenesis) were investigated in 70 invasive human breast carcinomas. Formalin-fixed, paraffin-embedded specimens were stained with monoclonal antibody against tenascin-C (DAKO and Biogenex). Anti-CD31 antibody (Biogenex), an acknowledged marker of stromal angiogenesis, was used to detect endothelial cells. Tenascin immunostaining was positive in the tumours around the persisting normal ducts, around tumour-cell nests, in the neostroma, in some tumour cells, and it was found in or around vascular channels. Tumour vascularity was assessed by quantitative vascular grading (Chalkley point count) and was related to the localization and intensity of tenascin immunoreactivity. 19 tumours (27.1%) were scored as low, 35 (50%) as medium, and 16 (22.9%) as having a high vascular grade. The positive correlation between the vascular grade and the tenascin immunopositivity in tumour stroma was observed. Our results suggest that tenascin expression may be associated with endothelial cell activation and may play an important role in tumour angiogenesis.

Sterically inhomogenous viscoelastic behavior of human saccular cerebral aneurysms.

To clarify the mechanism leading to the development and rupture of intracranial aneurysms, tensile strength and viscoelastic parameters of 22 human saccular aneurysms were investigated. Meridional and circumferential strips from the thin and the thick part of the aneurysm sack and 18 control strips from the basilar artery of 8 patients with pathologies not affecting the cerebral arterial system were studied. The length of the strips was increased in 200- microm steps, while distending force was recorded. Tensile strength and viscoelastic parameters were computed. In both directions, tensile strength of thick strips was significantly lower than that of controls. In the meridional direction, tensile strength of thin strips was significantly larger than that of thick ones (14.5 +/- 4.1 x 10(6) vs. 7.5 +/- 2.0 x 10(6) dyn/cm2, p < 0.05). In the circumferential direction, thin strips tore at lower strain values than thick ones (29 +/- 4 vs. 55 +/- 16%, p < 0.05). Viscoelastic parameters changed in parallel. In circumferential direction, values of thick and thin strips were significantly lower than those of controls. In the meridional direction, values of thin strips were significantly higher than those of the thick ones. These observations show that characteristic mechanical deterioration and steric inhomogeneities accompany the loss of smooth muscle cells and the derangement of connective tissue elements in the wall of intracranial aneurysms, which may explain certain steps in their initiation, enlargement and rupture.

Correlation between karyotype and ultrasound findings in patients with failed early pregnancy.

The aim of this retrospetive study was to identify any consistent patterns between ultrasound findings in failed pregnancies and either normal or abnormal karyotypes. The study involved 102 women in whom the diagnosis of early pregnancy failure was made sonographically. The criteria for a failed pregnancy were: a gestational sac with a mean diameter of > 12 mm without a yolk sac; a yolk sac of > 6 mm mean diameter with or without abnormal morphology that ultimately failed to develop an embryonic structure; and an embryo with a crown--rump length (CRL) of > 5 mm without cardiac activity, or the loss of previously identified cardiac activity. All patients underwent elective dilatation and curettage (D & C) and products of conception were sent for karyotyping. Forty-four pregnancies (43%) had abnormal karyotypes. Of these, 33 (75%) were trisomies. The other 11 included four triploidies, one tetraploidy, two with monosomy X, and four others (unbalanced complement, isochromosome, terminal deletion and translocation). Fifty-eight pregnancies (57%) had normal karyotypes, of which 52 were 46,XX and six were 46,XY. The furthest sonographic anatomic landmark achieved did not differ with respect to karyotypic findings. An abnormal yolk sac was found in 10/58 cases (17.2%) with normal karyotypes and 8/44 cases (18.2%) with abnormal karyotypes. There were eight cases of trisomy 16, of which only two manifested an embryonic structure, but neither of which had cardiac activity; the largest was 4 mm. There were four cases of trisomy 22, of which three developed embryos with a CRL of > 10 mm with cardiac activity (11, 11 and 18 mm, respectively). In three cases of mosaicism, embryos developed cardiac activity, and were 9, 19 and 16 mm. Two cases of monosomy X had embryos of 14 and 24 mm. Only one out of five cases with multiple trisomies developed to a point at which any embryonic structure was identifiable on ultrasound examination. The ultrasound appearance of early pregnancy failure in terms of furthest anatomic landmark reached was not significantly different in cases with normal or abnormal karyotype. An abnormally enlarged yolk sac, presumably secondary to hydropic change, is a non-specific finding of failed pregnancy, and did not correlate with karyotypic abnormality (trisomy 22, mosaic, monosomy X) seem to develop further prior to embryonic demise than those with certain others (trisomy 16, multiple trisomies and unusual other variants.

The role of progestationally regulated stromal cell tissue factor and type-1 plasminogen activator inhibitor (PAI-1) in endometrial hemostasis and menstruation.

The physiologic mechanisms whereby the human endometrium maintains hemostasis during endovascular trophoblast invasion, yet permits menstrual hemorrhage, are unknown. This paradoxical relationship was investigated by evaluating endometrial expression of tissue factor (TF), the primary initiator of hemostasis, and plasminogen activator inhibitor-1 (PAI-1), the primary inhibitor of fibrinolysis. We observed increased immunostaining for TF and PAI-1 in sections of decidualized stromal cells from luteal phase and gestational endometrium. To determine whether TF and PAI-1 expression are directly linked to decidualization, both endpoints were monitored in a well described in vitro model of decidualization. Thus, confluent stromal cell cultures were exposed to vehicle control, 10(-8) M estradiol (E2), 10(-8) to 10(-6) M medroxyprogesterone acetate (MPA) or both E2 + MPA for 2-24 days in serum-containing or defined media. The progestin enhanced the content of stromal cell-associated immunoreactive and functionally active TF and PAI-1 released into the medium and elevated levels of stromal cell TF and PAI-1 mRNA. While E2 alone was ineffective, it greatly augmented MPA-enhanced TF and PAI-1 protein and mRNA content. Dose-dependent effects on TF and PAI-1 content were observed between 10(-8) to 10(-6) M MPA +/- E2. Similar results were observed for decidual cells derived from first trimester endometrium and cultured in type 1 collagen gels. Following optimal induction of TF and PAI-1 expression by E2 + MPA in stromal cell cultures, removal of these steroids greatly reduced levels of both TF and PAI-1 protein and mRNA within 4 days. These studies suggest a mechanism whereby endometrial hemostasis is maintained during trophoblast invasion yet reduced at the end of nonfertile cycles to permit menses.

[Pulmonary and pleural reaction patterns to artificial mineral fibers --rockwool]. / Pulmonale und pleurale Reaktionsmuster auf künstliche Mineralfasern--Rockwool.

Epidemiological and animal studies about the fibrogenic and carcinogenic properties of natural mineral fibers required the development of man made vitreous fibers as substitutes for asbestos containing material. The question about the possible fibrogenic and carcinogenic properties of man-made vitreous fibers is not yet answered. By means of different experimental animal studies we tried to investigate the man made vitreous fibers-related pulmonary and pleural diseases. The experimental administration of rockwool induce lesions in the lung of the rats. The lung showed an extensive granulomatous inflammation. In the observation time of 10 months we did not find any malignant tumor of the lung and the pleura.

Cellular autofluorescent pigment and interstitial fibrosis in smoker's lung.

The mechanisms by which cigarette smoking lead to bronchopulmonary diseases are incompletely understood. The most characteristic lesion is a chronic macrophage-alveolitis accompanied by slight fibrosis and emphysema. The macrophages contain a ceroid-like granular autofluorescent pigment in their lysosomes. Using immunohistochemical methods, open lung-, transbronchial biopsies and cells obtained by broncho-alveolar lavage from cigarette smokers were studied: anti-human macrophage serum and anti-human elastase, immune sera against type I, type III collagens and fibronectin were used in the demonstration of the cellular components of alveolitis and the connective tissue constituents of fibrosis. The characteristic red-brown autofluorescent pigment of the macrophages was also found in an extra-alveolar location mainly in peribronchial, septal and pleural scars. Similar emission colour occurred focally in the elastic laminae of fibrotic alveoli and sclerotic arteries. Granular fluorescent pigment was found in many bronchial epithelial cells. The epithelial pigmentation was associated with increased transcription of nucleic acid proteins, revealed by colloid silver (AgNOR) reaction. The results suggest that the autofluorescent pigment substances in macrophages may indicate or also play a role in the development of pathological connective tissue and epithelial changes of smoker's lung, in addition to the known mediators and enzymes.

Changes of DNA-acridine orange binding in monocytes and endothelial cells of hypertensive arteries.

The interactions of monocytes and endothelial cells were examined employing acridine orange (AO) binding to chromatin as an electron microscopic probe for studying changes in nuclear activity. The number of AO labeled nuclei were determined in adherent and sub-endothelial monocytes, and endothelial cells either associated with or devoid of monocytes in mesenteric arteries of renal hypertensive and normotensive rats. No AO-positive cell nuceli were found in the few adherent monocytes in the normotensive rats while in hypertensive rats 63% of the adherent monocytes and 86% of the subendothelial monocytes were labeled by AO. The number of AO-positive cell nuclei increased in the endothelium from 2% in normotensive rats to 17% in hypertensive animals. Fifty-seven percent of these endothelial cells were associated with monocytes adhering to their surfaces. Thirty-nine (52%) of AO-positive and 24 (52%) of AO-negative cell pairs represented 84% of cell pairs with identical nuclear activity. These findings indicate a sequential interaction of monocytes and adjacent endothelial cells recognized at the nuclear level. According to other experimental work the binding capacity of chromatin for AO increases as cell nuclei are reactivated by stimulation of cell proliferation. In addition, AO labeling of chromatin visualized electron microscopically is confined to three of the four stages of the cell cycle. Therefore AO labeling in the present experiments might indicate the reentrance of monocytes and endothelial cells into the cell cycle.

Passive geometric and elastic properties of human cadaver common carotid artery segments after intraluminal enzyme digestion with the aid of a four-way double balloon catheter.

A combined intraluminal nonenzymatic-enzymatic digestion procedure was carried out in situ on the common carotid arteries of 21 human cadavers with the aid of a four-way double balloon catheter. Denaturing pretreatment with NaOH, 2.5 mol for 20 min followed by crude pancreatic extract, 20 mg/ml for 20 min were used. Mechanical properties of cylindrical segments were measured in vitro, and compared with contralateral control segments. The inner radius increased and wall thickness decreased significantly at each pressure level in the 0-200 mmHg (0-26.7 kPa) intraluminal pressure range. Volume of wall material per unit length of the segment decreased by 11.6%, cross-section of the lumen at 100 mmHg (13.3 kPa) intraluminal pressure increased by 13.6%. These results show that the internal layers of human arteries can be removed by enzyme digestion increasing the lumen, while intact (medial and adventitial) layers still can give sufficient strength to the arterial wall.

[The effect of endothelial interaction on the binding capacity of DNA acridine orange in hypertension]. / A monocyta endothel interakció hatása a DNS akridin-oranzs kötöképességére hypertoniában.

Interaction of monocytes and endothelial cells were examined by acridine orange reaction in arteries of normotensive and hypertonic rats. The method is suitable for electron microscope study of DNA template activity. Activity of gen was compared in surface bound and subendothelial and in nonadhesive endothelial cells bound to monocyte. Acridine orange positivity, indicative of genic activity, was not found in few monocytes adhered in arteries of normotensive animals, while 62% of adhesive monocytes of hypertonic and 86% of ones accessed to endothelial space contained the products of acridine-orange-chromatin reaction in its nuclear, showing with it the gene derepression quickly ensuing in acute hypertensive vascular lesions. Only 2% of endothelial cells of pseudo-operated normotensive animals showed acridine orange positivity, while 17% of endothelial cells of hypertonic animals were positive. In the latter animals, 57% of endothelial cells showed acridine orange positivity, if monocytes adhered to their surface. In 84% of monocyte-endothelial pairs connected to each other, the nuclei have activity of same sign. Results show intensive activation of genes of monocytes in relation with adhesion and migration to vascular wall. Presumably, both hypertonia and monocyte adhesion have contributed to the increase of template activity of endothelial cells.

Comparison of the effects of NaF and CaF2 on rat gastric mucosa. A light-, scanning- and transmission electron microscopic study.

Rat stomachs were studied after intragastrically administered, repeated doses of 0.1 N HCl, 100 mmol/l NaF, 50 mmol/l CaF2 in 0.1 N HCl, respectively. NaF produced extensive desquamation and cell injury, while CaF2 caused some desquamation and a slight decrease in secretory activity as revealed by light microscopic, SEM and TEM examinations.