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.

Follow-up of prostate carcinoma patients treated with total androgen blockade by repeated map-biopsy.

A prospective study was carried out on ten patients with prostate cancer. The TNM stage, Gleason's grade, general clinical status and, serum PSA level were all registered by the time of diagnosis. Total androgen blockade (TAB) was performed. Serum PSA control and general clinical examination re-biopsy was performed on average 107 days after the start of the therapy. The pre- and post treatment histology included HE and Tunel reaction to show apoptotic cells, as well as p53, bcl2 and Ki67 immunostaining. Clinical improvement of the disease, manifested by regression or by steady state was observed in all ten patients. An increase of apoptotic index, and a decrease of mitotic index was detected in most cases. The serum PSA level decreased in all patients. Ki67, bcl2 and mutant p53 were strongly expressed in tumor cells of patients in whom Gleason's grade was 7 or higher and decreased markedly in all cases upon therapy. Data obtained by repeated biopsy in the course of TAB therapy are indicators of the effectiveness of TAB, like changes in serum PSA, and may be considered as predictive factors.

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.

Bioaccumulation of lead by the lichen Acarospora smaragdula from smelter emissions.

Accumulation of lead in the crustose lichen Acarospora smaragdula sensu lato is reported in the vicinity of an ore- processing plant where it is subjected to acidification and metal particulate fallout. A combination of light microscopy, X-ray element mapping, field emission scanning electron microscopy (FESEM) and other analytical techniques identifies Pb accumulation within specific fungal tissues derived from smelter particles (PM10s). No Pb was detected within the photobiont layer. Our studies suggest that Pb is highly mobile under the prevailing acidic conditions, and is fixed within the lichen cortex and melanized apothecia. Lead is also accumulated within the medulla and at the rock-lichen interface where it may precipitate as amorphous botryoidal encrustations on medullary hyphae and iron-rich particles. Modern FESEMs and microprobes enable analysis of minute quantities of material, and are important tools in understanding the fate of metals within lichens necessary to develop their use as predictive and sensitive bioindicators of aerial particulate contaminants. We suggest that crustose lichens, hitherto largely ignored in metal pollution studies, may be useful bioindicators of aerial particulate contaminants in polluted areas where macrolichens are absent.

[Alpha-1-antitrypsin deficiency and liver diseases]. / Alfa-1-antitripszin-hiány és májbetegség.

Authors studied liver biopsy of hundred infants and young children. 15 livers were normal and three of 85 patients suffering from liver disease had liver changes associated with serious alpha--1-antitrypsin (AAT) deficiency (in two cases with portal fibrosis associated with inflammation, in one case with portal fibrosis associated with moderate active inflammation of reduction of intrahepatic biliary ducts). Typical positive, diastase resistant globules were in the hepatocytes of only one patient, but AAT could be demonstrated in periportal liver cells of all three patients by AAT specific immuneperoxidase method. Of 830 hepatic cirrhosises in adult age, in eight cases PAS positive, diastase-resistant AAT immunreactive globules occurred in periportal hepatocytes, suggesting AAT deficiency; however, the AAT level and fenotype of these patients were not known. Examination of possibility of AAT deficiency should be performed in every case, where the cause of liver disease is unsolved; this examination is especially indicated by the presence of typical PAS positive, diastase-resistant, AAT immunreactive globules in hepatocytes.

[Liver changes suggesting alpha-1-antitrypsin deficiency (a neglected disease)]. / Alpha-1-antitrypsin-hiányt sugalló májelváltozás (egy kevés figyelemre méltatott ártalom).

Liver needle biopsies of one hundred infants and children were examined. In fifteen of them the liver was normal. Of the 85 patients with liver disease three had liver changes due to severe alpha-1-antitrypsin (AAT) deficiency. In two cases fibrosis and inflammation of the portal fields could be seen. In the third case in addition to portal fibrosis and moderate periportal inflammation paucity of the intrahepatic bile ducts was found. Characteristic PAS-positive diastase resistant hepatocellular globules occurred only in one case but, using immunoperoxydase method, periportal hepatocytes showed AAT positivity in all three infants. Of 830 adult patients with liver cirrhosis 8 had PAS-positive diastase resistant AAT immunoreactive globules in the periseptal hepatocytes suggesting AAT deficiency, however, the serum AAT level and the phenotype of them were unknown. Investigation for AAT deficiency should be carried out in children and young adults with a history of neonatal liver disease and possibly in all patients with liver disease of uncertain etiology, especially in those with PAS-positive, AAT immunoreactive hepatocellular globules.

[Ultrastructure of Ito cells in fibrotic liver diseases]. / Az Ito sejtek ultrastruktúrája fibrosissal járó májbetegségben.

Authors have studied the ultrastructure of Ito-cells in percutane needle-biopsy samples of hepatic tissue from 36 patients. In 3 samples no alteration was found, in four only moderate steatosis occurred without fibrosis. In four samples portal, in ten centrilobular and in five periportal fibrosis was detected, while in five samples cirrhosis was pointed out. In the intact liver, Ito-cells occur not only in the spaces of Disse but also in the wall of the central vein. In portal fibrosis Ito-cells were similar in location and structure to those in the non-fibrotic liver. In centrilobular and periportal fibroses as well as in cirrhosis Ito-cells in the fibrotic parenchyma areas were either localized in the accumulated connective tissue or at the border of connective tissue and hepatocytes. These interstitial Ito-cells contained small amount of lipid, abundant dilated rough endoplasmic reticulum filled with a flocculent material, well-developed Golgi-complex and often bundles of 5 nm thick filaments with dense territories. Close to the Ito-cells immature collagen-fibrils and basement membrane-fragments were seen. A close relationship was pointed out between activated Ito-cells and lymphocytes. In parenchyma areas away from fibrotic foci the structure of Ito-cells was comparable to that of Ito-cells in intact hepatic lobules. Observations suggest that Ito-cells are related to fibroblasts and myofibroblasts and play a role in the pathogenesis of fibrosis occurring in human liver diseases.

Ultrastructure of the Ito cells in liver disease associated with fibrosis.

The ultrastructure of Ito cells was examined in percutaneous needle liver biopsies of 36 patients. In three cases the liver was normal. Four specimens showed mild or moderate fatty degeneration. In the rest of the liver samples various types of fibrosis were seen: in four cases portal, in ten centrilobular, in eight periportal fibrosis and in five cases cirrhosis. In the normal liver Ito cells occurred not only in the Disse spaces but also in the walls of the terminal hepatic venules. In livers showing portal fibrosis the ultrastructure of Ito cells was similar to that seen in the normal liver. In the fibrotic areas of liver samples showing centrilobular or periportal fibrosis or cirrhosis Ito cells localized inside the fibrotic tissue and along the border between the connective tissue and hepatocytes. These interstitial Ito cells contained few lipid, abundant rough endoplasmic reticulum with dilated tubules filled by a flocculant material, well developed Golgi complexes and often bundles of 5 nm thick filaments with densities. These cells also known as activated Ito cells were in places surrounded by immature collagen fibrils and basement membrane fragments. There was a close contact between activated Ito cells and lymphocytes. The ultrastructure of Ito cells localizing in non-fibrotic areas did not differ from that seen in normal liver lobules. These observations suggest that Ito cells are related to fibroblasts and myofibroblasts and play an important role in the pathogenesis of liver fibrosis in humans.

A case of liver cirrhosis with alpha-1-antitrypsin globules and hepatitis B surface antigen seropositivity.

PAS staining, immunohistochemical examination and electron microscopy revealed presence of alpha-1-antitrypsin (AAT) globules in the hepatocytes of a HBsAg and anti-HBc seropositive female patient diseased of liver cirrhosis. The possible causes of cirrhosis are briefly analysed and the diagnostic importance of PAS-positive, amylase-resistant hepatocellular inclusions is discussed. Apart from the case reported, only two of 509 cirrhotic livers of adults, examined either by biopsy or post mortem, demonstrated similar characteristic PAS-positive globules. This indicates that in the population group (135,000 persons) referred for health care to the hospital where the examinations were done, AAT deficiency has played a negligible role in the development of liver cirrhosis in adults.

Investigations on blood and cerebrospinal fluid lymphocytes in patients suffering from tick-borne encephalitis.

This paper deals with the immuno-pathological characteristics of tick-borne encephalitis. With the appearance of neurological signs, there is a strong T cell reaction in th peripheral blood. The following seven days are characterized by the appearance of specific cell-mediated reaction in the peripheral blood; in the cell-mediated reaction in the peripheral blood; in the cerebrospinal fluid, an increasing number of B cells and specific antibodies can be detected. After a week, the percent positivity of cell-mediated immune reactions is higher and there is a T cell dominance in the cerebrospinal fluid. Intact cellular immunity against specific antigen is required for the management of this disease. There are major differences in the results obtained from patients suffering from meningitis alone and those suffering from meningoencephalitis.

Changes in transcription of endogenous type-C virus genome during mouse liver regeneration.

In the present study the extent of endogenous, type-C virus genome transcription in normal and regenerating mouse liver was analysed by using the technique of nucleic acid hybridization. The RNA preparations from regenerating liver tissues collected at various intervals following partial hepatectomy, and from normal liver samples of BALB/c mice, were hybridized to 3H-DNA complementary to 60 to 70S RNA of an endogenous, N-tropic virus, released spontaneously from BALB/c mouse cells in culture. Although partial transcription of the endogenous virus genome can be clearly detected in normal liver, a significant increase in the level of virus-specific RNA synthesis in the regenerating liver, in comparison to normal liver, is apparent, following partial hepatectomy. This increase in virus-specific RNA synthesis attains its highest level just before the level of DNA synthesis in the regenerating liver reaches its maximum. These observations may indicate a qualitative or quantitative change in the endogenous type-C virus genome transcription pattern in hepatocytes, in response to partial hepatectomy and suggest that this change in the transcription pattern and the initiation of cell proliferation, in regenerating livers, are probably sequential and related events.

Multiple structures of adeno-associated virus DNA: analysis of terminally labeled molecules with endonuclease R-Hae III.

The double-stranded form of adeno-associated virus (AAV) DNA has about 20 sites sensitive to endonuclease R.Hae III from Haemophilus aegypitus; the fragments produced fall into about 13 size classes, 8 of which contain single fragments. The location of the Hae III-produced AAV fragments relative to the three EcoR1 fragments was determined. Using revised figures for the molecular weights of the Hae III cleavage products of phiX174 replicative form DNA, we calculated that AAV DNA contains about 4,000 nucleotides. After Hae III digestiion of duplex DNA terminally labeled with 32P using polynucleotide kinase, the majority of fragments containing a 5' 32P label were about 40 nucleotides in length, and fragments of similar size were generated from each end, suggesting that the Hae site closest to the end is within the terminal repetition. Two more-slowly-migrating cleavage products also bore 5' 32P end label. These three terminally labeled species were also generated from single-stranded AAV DNA by digestion with Hae III, and evidence that one may have a nonlinear ("rabbit-ear") structure is presented. The predominant 5' terminal base was identified as thymine for both the plus and minus strands of AAV. Single-stranded AAV molecules could not be efficiently covalently circularized by incubation with polynucleotide ligase or ligase plus T4 DNA polymerase.

Site of cleavage of superhelical phiX174 replicative form DNA by the single strand-specific Neurospora crassa endonuclease.

Experiments with the Neurospora crassa single strand-specific endonuclease have provided evidence for the existence of regions of partially single-stranded character in covalently closed superhelical replicative form DNA of phiX174. The nuclease converts the superhelical molecules to either singly hit relaxed circular or doubly hit linear molecules. We show that the initial cleavage of phiX174 superhelical DNA is a "nick" bounded by a 5'-phosphate and a 3'-hydroxyl; no nucleotides are excised as evidenced by the ability of T4-polynucleotide ligase to reform the phosphodiester bond. The nick can be found in either strand of the double-stranded DNA and is either randomly distributed or at least can be found at any one of many possible locations in the genome. Thus, the regions in phiX174 superhelical molecules that are sensitive to the N. crassa nuclease do not occur at highly specific sites in the genome.

Mechanism of replication of single-stranded PhiX174 DNA. VII. Circularization of the progeny viral strand.

Linear phiX174 single-stranded DNA can be isolated from phiX phage particles produced under various conditions. About half of the linear strands have a dGMP residue at the 5' end, the remaining have roughly comparable amounts of dCMP, dTMP, and dAMP. The linear strands can be converted to covalently closed circular molecules by polynucleotide ligase, but only after they have been incubated with T4 DNA polymerase and deoxynucleoside triphosphates. Experiments with endonuclease R, the restriction enzyme from Haemophilus influenzae, indicated that the nucleotides incorporated into the DNA during this reaction were found predominantly in a limited region of the genome. The results suggest that the normal intermediate in single-stranded phiX174 DNA synthesis may be a single-stranded linear molecule which is shorter than unit length and is intrinsically capable of circularization.