Mladen Petrovečki

Prognostic Parameters for Survival of Patients with Malignant Mesenchymal Tumors of the Uterus

Malignant mesenchymal uterine neoplasms are the most aggressive type of primary uterine tumors, with most patients dying within a few years of diagnosis. Thus, it would be very important to define prognostic factors for predicting the malignancy potential of at least some of their subtypes.

Indirect demonstration of the lifetime function of human thymus

The aim of this study was to test the hypothesis that human thymus maintains its function as the site of early T cell development throughout life, but to a progressively diminishing extent. Mononuclear cell suspensions prepared from the samples of 39 human thymuses were analysed for the total number of cells per gram of thymus tissue, percentage of single marker‐positive CD2, CD4 and CD8 cells, percentages of double‐positive CD4 CD8 and CD2 CD8 cells, double‐negative CD4 CD8 cells, absolute numbers of these cells per gram of tissue, and extent of the in vitro proliferation upon stimulation with concanavalin A (Con A), phytohaemagglutinin (PHA) and pokeweed mitogen (PWM) mitogens. The main outcome measures were flow cytometric data on thymus lymphoid cell composition (according to CD classification), expressed as percentages and numbers of cells per gram of thymus tissue. The total number of mononuclear cells expressed per gram of thymus tissue exponentially decreased with age. The slope of none of the analysed cell subpopulations differed from the slope of the line constructed for age‐related decline of the total number of mononuclear cells (−0.024 on a semilogarithmic scale). The thymuses of all ages contained all analysed cell subpopulations in approximately the same proportions: percentages of these cell subpopulations did not change with age, except for all CD4+ (P = 0.017) and double‐positive CD4+ CD8+ (P = 0.016) cells, which tended to decrease with age. The extent of proliferation of thymus cells upon stimulation with T and B cell mitogens was unrelated to age. We conclude that the thymus retains its function as the site of differentiation of T lymphocytes throughout life. With respect to the number of involved lymphoid cells, the function exponentially decreases with age.

Age‐dependent haematological disturbances in anaesthetic personnel chronically exposed to high occupational concentrations of halothane and nitrous oxide

Anaesthetic staff chronically exposed to high occupational concentrations of halothane and nitrous oxide were tested for numerous haematological and cellular function parameters at the peak of the working season and after 3 weeks vacation. The analysis of data was performed to compare differences in subjects younger and older than the age of 40 years, respectively when compared with normal controls. The analysis revealed a higher recovery of erythrocyte’count in the blood of older staff, and stronger disturbance of leucocyte formation in younger staff. In contrast, monocytes appeared to be more stable in the younger staff as were the T and B lymphocyte counts. After stimulation with PHA, Con A and PWM mitogens, lymphocytes from the older age group incorporated a significantly higher amount of tritiated thymidine, but stimulation indices did not differ. Natural killer cell numbers appeared equally affected; natural killer cell activity was unaffected, but there was an increase in activity in the younger staff after the vacation. Serum immunoglobulin concentrations tended to be more affected in older individuals at the peak of the working season.

Hypersensitivity to Parietaria officinalis pollen in newcomers to the area with the plant

Hypersensitivity to Parietaria officinalis (wall pellitory) pollen and other environmental allergens was studied in pollinosis patients allergic to P. officinalis pollen who were born in areas without P. officinalis and later moved to the city of Split, where P. officinalis is responsible for some 65% of pollinosis cases. Highly significant positive correlations were found for both the intensity of skin test reaction and concentration of specific serum IgE with the length of residence in the area. In contrast, the respective data on subjects hypersensitive to P. officinalis pollen allergen, but born and living in the area of Split, revealed a tendency to negative correlation between age and intensity of hypersensitivity to P. officinalis. A number of patients from both groups were tested for presence of serum IgE antibodies specific for 14 common environmental allergens. Hypersensitivity to P. officinalis pollen was associated with hypersensitivity to olive, mugwort, and birch pollen in newcomers; hypersensitivity to birch and, to some extent, olive pollen was significantly more frequent in newcomers than in autochthonous patients who were allergic to P. officinalis pollen. Regardless of whether the patients were autochthons or newcomers to the area with P. officinalis, hypersensitivity to P. officinalis mostly excluded hypersensitivity to Dermatophagoides farinae and D. pteronyssinus, and vice versa.

Correlation of two methods for determination of cathepsin D in breast carcinoma (immunohistochemistry and ELISA in cytosol)

We have studied the correlation of two methods(immunohistology and ELISA in cytosol) of cathepsin D(CD) determination in breast carcinoma patients. Fifty sixspecimens of tumor tissue were collected consecutively, andCD expression in tumor tissue and tissue macrophageswas determined by standard immunohistochemistry using the aNCL-CDmanti-cathepsin D mouse monoclonal antibody (Novocastra Laboratories Ltd.,Newcastle, UK). Additionally, CD concentration was determined byELISA in cytosol of the same breast carcinomaspecimens. CD positivity was correlated with tumor size,histological grade of tumor, and the cytosol progesteroneand estrogen receptor concentrations. There was no statisticallysignificant correlation between examined parameters and either CDpositivity by immunohistochemistry or cytosol CD concentration. Thecorrelation between CD expression in tumor cells ofbreast carcinoma by immunohistochemistry and cytosol CD positivitywas not found either. However, there was asignificant association between abundance of CD positive stromalmacrophages and cytosol CD concentration in all histologicaltumor types (p < 0.05). CD positive macrophageswere abundant in most of cytosol CD positivespecimens.These results suggest that breast cancer cytosol CDconcentration is the cumulative result of CD contentin both carcinoma cells and stromal macrophages.

Effects of Isolation and Inbreeding on Human Quantitative Traits: An Example of Biochemical Markers of Hemostasis and Inflammation

Abstract Isolation is a known force in evolutionary biology and one of the main factors in speciation. One of the main consequences of severe isolation is reduced mate choice, which results in the occurrence of inbreeding as a result of isolation. We investigated the effects of individual genome-wide heterozygosity measured as the multilocus heterozygosity (MLH) on biochemical markers of hemostasis and inflammation in 1,041 individuals from the island of Vis, Croatia, where inbreeding is prevalent and a wide range of variation in the genome-wide heterozygosity is expected. Assessment of individual genome-wide heterozygosity was based on genome-wide scans using 800 microsatellite (STR) and 317,503 single nucleotide (SNP) polymorphic markers in each examinee. In addition, for each examinee we defined a personal genetic history (PGH) based on genealogical records. The association between PGH and MLH and fibrinogen, D-dimer (Dd), von Willebrand factor (vWF), tissue plasminogen activator (tPA), and C-reactive protein (CRP) was performed with a mixed model, controlling for possible confounding effects. PGH was a significant predictor only for tPA (P < 0.001), whereas neither of the two MLH measures exhibited significant association with any of the investigated traits. The effects of individual genome-wide heterozygosity are most likely expressed in highly polygenically determined traits or in traits that are mediated by rare and recessive genetic variants. Weak associations between PGH and MLH and markers of hemostasis and inflammation suggest that their genetic control may not be highly polygenic and that they could be promising targets for genetic association studies.

Prognostic significance of cytochemical analysis of leukemic M2 blasts

Cytochemical analysis of leukemic blasts from 46 patients with acute myeloblastic M2 leukemia (according to the FAB classification) was performed before and after cytostatic therapy, and compared with findings obtained in 20 age- and sex-matched control subjects. Cytochemical findings for myeloperoxidase (MPO), Sudan black B, acid phosphatase and alpha-naphthyl-acetate esterase (ANAE) were related to the achievement of the first complete remission (CR),i.e. data were compared after the patients had been divided into CR and non-CR groups. The analysis clearly showed that a high proportion of myeloperoxidase- and, to a lesser extent, Sudan black B-positive blasts before treatment may have constituted a significantly unfavourable prognostic factor.

Prognostic significance of cell cycle parameters in infiltrative ductal breast carcinoma

Flow‐cytometric DNA analysis was performed retrospectively from paraffin‐embedded blocks in 158 consecutive ductal infiltrative breast carcinoma patients grades I–III. Normal breast tissue was used as control. Tumor proliferative activity, cell ploidy, and DNA index (DI) were related to age of patients, histological grade of tumor, tumor size, axillary lymph node status, estrogen and progesterone receptor status, menopausal status, TNM clinical classification, and survival. There was a significant association between DNA aneuploidy and a high cellular proliferative activity, increased DI, poor differentiation of tumor, primary tumor size, number of positive lymph nodes, and postmenopausal state. Increased proportion of cells in S‐phase was associated with positive lymph node status and higher number of positive lymph nodes. The cell cycle parameters had no prognostic value either for overall survival of disease‐free survival of the patients. J. Clin. Lab. Anal. 12:131–136, 1998.

Granulocyte-monocyte colony forming unit content of autologous bone marrow transplants in patients with haematological malignancy.

Cell viability and number of granulocyte-monocyte colony forming units (CFU-GM) were systematically assessed in 57 patients who had undergone transplantation of the autologous bone marrow for treatment of haematologic malignancies. Bone marrow cell cultivation in agarose with feeder layers appeared inferior to that performed in agarose with recombinant human granulocyte-monocyte colony stimulating factor and methylcellulose with phytohaemaglutinin leukocyte-conditioned medium. Since the transplant cells were frozen in liquid nitrogen between harvesting and reinfusion, the following samples were tested: buffy coat cells, buffy coat cells immediately after addition of dimethylsulphoxide, cell sample that had been frozen for 24 hours, and frozen transplant cells at the time of thawing and transplantation. Each procedural step decreased both cell viability and the number of CFU-GM, but since the lymphohaematologic recovery in all patients followed the pattern reported in the literature for high-quality transplants, we concluded that our transplants retained the necessary number of progenitor cells. It appears that the best strategy for dynamic assessment of the transplant quality would be to perform tests after every step of the transplant processing. Cell viability and number of progenitors per body weight in transplants were also found to be associated with probability of neutrophil reconstitution after bone marrow reinfusion.