Monika Korabiowska

Severe granulomatous allergic tissue reaction after hyaluronic acid injection in the treatment of facial lines and its surgical correction.

Hyaluronic acid is considered to be nonimmunogenic. Frequently, it is used for the correction of facial lines. It is believed that hyaluronic acid injection fillers are safe and have no occurrence of serious adverse reactions or allergic reactions. Nevertheless, recent publications have documented the rate of intermittent swelling and severe granulomatous allergic reactions that evolved into abscesses. A clinical case of a 54-year-old patient is presented. After injection of hyaluronic acid in the treatment of nasolabial folds elsewhere, she developed palpable painful erythematous nodules evolving into abscesses several month after injection. Surgical treatment and correction of these lesions after hyaluronic acid injection of the nasolabial folds and the histological findings of these erythematous nodules are described. Histological and clinical examination documented intermittent swelling and severe granulomatous allergic reactions that may render the use of hyaluronic acid unacceptable. Patients should be informed of the potential complications when treating facial lines with hyaluronic acid gel.

Disruption of the Pelota Gene Causes Early Embryonic Lethality and Defects in Cell Cycle Progression

ABSTRACT Mutations in either the Drosophila melanogaster pelota or pelo gene or the Saccharomyces cerevisiae homologous gene, DOM34, cause defects of spermatogenesis and oogenesis in Drosophila, and delay of growth and failure of sporulation in yeast. These phenotypes suggest that pelota is required for normal progression of the mitotic and meiotic cell cycle. To determine the role of the pelota in mouse development and progression of cell cycle, we have established a targeted disruption of the mouse Pelo. Heterozygous animals are variable and fertile. Genotyping of the progeny of heterozygous intercrosses shows the absence of Pelo −/− pups and suggests an embryo-lethal phenotype. Histological analyses reveal that the homozygous Pelo deficient embryos fail to develop past day 7.5 of embryogenesis (E7.5). The failure of mitotic active inner cell mass of the Pelo −/− blastocysts to expand in growth after 4 days in culture and the survival of mitotic inactive trophoplast indicate that the lethality of Pelo-null embryos is due to defects in cell proliferation. Analysis of the cellular DNA content reveals the significant increase of aneuploid cells in Pelo −/− embryos at E7.5. Therefore, the percent increase of aneuploid cells at E7.5 may be directly responsible for the arrested development and suggests that Pelo is required for the maintenance of genomic stability.

Downregulation of the Retinoblastoma Gene Expression in the Progression of Malignant Melanoma

The retinoblastoma gene is a cell cycle regulator preventing cells from entering into S-phase. An altered expression of the retinoblastoma gene has been reported in the majority of human malignancies. The main aim of this study was to investigate retinoblastoma gene expression in the full spectrum of melanoma progression from naevus to melanoma metastases by applying immunohistochemistry and RT-PCR. All naevi with and without dysplasia showed high expression of the retinoblastoma gene. In primary melanomas, Rb-positive cells were found in 82 out of 106. Loss of expression correlated with an increase in Clark level and shorter survival rates. An independent prognostic role of the retinoblastoma gene was confirmed by Cox multivariate analyses (p < 0.01). In melanoma metastases, retinoblastoma gene expression (at the RNA level) was found in 18 out of 26 melanoma lymphatic metastases, and in 2 out of 5 liver metastases. Our results indicate a downregulation of the retinoblastoma gene in the progression of melanocytic tumours.

Analysis of adenomatous polyposis coli gene expression, APC locus-microsatellite instability and APC promoter methylation in the progression of melanocytic tumours

Adenomatous polyposis coli gene (APC) defects have been demonstrated for the first time in familial adenomatous polyposis. Recent reports indicate that the APC gene is an intermediary between cell adhesion molecules and the cytoskeleton and that it may function as a gatekeeper of colonic epithelial proliferation. The objective of this study was to analyse APCs presence in lentigos, primary melanomas and melanoma metastases. By immunohistochemistry, APC was demonstrated in all lentigos, in 75 out of 88 primary melanomas and in 16 out of 28 melanoma lymphatic metastases. The percentage of immunolabelled tumour cells (APC index) in lentigos ranged between 5 and 69%, in primary melanomas between 0 and 98% and in melanoma metastases between 0 and 52%. Statistically significant differences between lentigos and primary melanomas and between lentigos and metastases in APC expression were found. In a multivariate analysis, APC showed an independent prognostic impact. Analysis of microsatellite instability in the APC locus was performed on 29 melanomas. Microsatellite instability was found in 5/29 melanomas and loss of heterozygosity in 1/29 melanomas. Promoter methylation of APC was found in 6/10 APC-negative primary melanomas and in 9/10 APC-negative melanoma lymphatic metastases investigated. We conclude about important role of APC alterations for melanoma progression.

Down-regulation of Ku 70 and Ku 80 mRNA expression in transitional cell carcinomas of the urinary bladder related to tumor progression.

DNA-dependent protein kinase (DNA-PK) containing the regulatory subunits Ku 70 and Ku 80 plays a prominent role in the repair of double-stranded DNA breaks by a nonhomologous end-joining pathway maintaining genomic stability. In an attempt to elucidate the significance of the DNA-PK complex for human urothelial carcinogenesis, the expression of Ku 70 and Ku 80 was studied in 71 transitional cell carcinomas (TCC) of the urinary bladder of various grades and stages, and in relation to lifestyle and occupational bladder cancer risk factors. To analyse the mRNA expression of Ku 70 and Ku 80, real-time quantitative reverse transcription-polymerase chain reaction was used and the protein expression assessed by immunohistochemistry. Advanced high-grade, high-stage TCC expressed the mRNA of Ku 70 and Ku 80 at a lower level than superficial low-grade, low-stage carcinomas, suggesting down-regulation of the Ku system to be associated with progression of bladder cancer from a low to a high malignant potential. The protein expression of Ku 70 and Ku 80 was closely related and decreased consistently with increasing grades and stages, paralleling the expression of the mRNA. Among hazardous environmental bladder cancer risk factors, heavy consumption of coffee was associated with a twofold decreased Ku 70 and Ku 80 mRNA expression, whereas tobacco smoke did not substantially affect the activity of the Ku system, except for a trend towards a dose-response relationship in the expression of Ku 70 mRNA. There is some evidence that exposure to polycyclic hydrocarbons, paints and lacquer, and stone dust may modify the expression of Ku 70 mRNA. Although the underlying molecular genetic pathways are not yet clearly understood, our data indicate that down-regulation of the Ku system promotes progression of urothelial carcinogenesis to a more malignant and aggressive clinical behavior, presumably as a result of an impaired capacity for DNA repair.

Borderline Tumors of the Ovary : A Clinico-Pathologic and Immunohistochemical Study of 54 Cases

Objective:To study EGF‐R, HER‐2/neu (p185), p53, Mib‐1 (Ki‐67), Bax, Bcl‐2, ras expression and ploidy in borderline tumors of the ovary by assessing their frequency, and relationship to histologic type, tumor recurrence and survival.

Relation between DNA ploidy status and the expression of the DNA‐mismatch repair genes MLH1 and MSH2 in cytological specimens of melanoma lymph node and liver metastases

DNA‐mismatch repair is essential for preventing genetic instability, and its important protective role has been demonstrated in several tumors. The main aim of this study was to investigate the expression of MLH1 and MSH2 (on the RNA level) in melanoma liver and lymph node metastases, and to define the relation between DNA ploidy status and mismatch repair gene expression.

Expression rate of vinculin isoforms in human aortocoronary saphenous vein grafts

Aortocoronary bypass conduits derived from saphenous veins usually develop diffuse intimal thickening, one of the major causes of haemodynamically relevant graft stenosis. To elucidate the role of smooth muscle cell proliferation in late graft failure, specimens from highly stenotic or occluded vein grafts implanted into the arterial circulation for more than 5 years were tested for their expression rate of meta-vinculin. Since the cytoskeletal protein meta-vinculin is present exclusively in contractile smooth muscle cells, the determination of the relative amounts of meta-vinculin (150 kDa) and its low-molecular weight isoform vinculin (130 kDa) allows characterization of the phenotypic status of smooth muscle cells. Using immunoblotting techniques, the quantitative relation of meta-vinculin in tissue samples obtained from autoptic vein grafts (n=10) was measured and compared with those of native saphenous veins (n=6). In vein grafts, the fractional meta-vinculin content of the total vinculin immunoreactivity ranged from 32%-46% (mean 39.6%), whereas the range was 39%-53% (mean 46.7%) in native veins. By applying Students t-test, a statistical significance was not demonstrated suggesting that the majority of smooth muscle cells in intimal thickenings consisted of a contractile phenotype. Immunohistochemically, the vinculin immunoreactivity in the intimal layer of vein grafts was reduced as compared to native saphenous veins. The distribution of vinculin in grafted veins closely resembled that in arteriosclerotic coronary arteries with intimal thickening. Hence, our biochemical data demonstrate parallels between the pathogenesis of late vein graft stenosis and degenerative arteriosclerotic lesions.

Relationship between DNA ploidy-related parameters and the deletions in mismatch repair genes MLH1 and MSH2 in lentigo maligna and malignant melanomas

Abstract Defects in DNA mismatch repair genes MLH1 and MSH2, first described in hereditary nonpolyposis colon cancer (HNPCC), have been postulated to be responsible for malignant transformation in several tumours. To date there are no data on cutaneous tumours. Using a PCR assay it was possible to identify deletions in MSH2 (exonic regions 12 and 13) in 16 of 47 lentigos maligna and in 10 of 36 malignant melanomas. Deletions in MLH1 (exonic regions 15 and 16) were found in 11 of 47 lentigos and in 15 of 36 melanomas. Comparison of DNA ploidy-related parameters between lentigos with and without exonic deletions in MSH2 and MLH1 did not show any significant differences. In contrast, melanomas positive and negative for exons 12 and 13 (MSH2) (26/36 and 10/36, respectively) differed significantly with respect to the percentages of diploid cells ( P = 0.0179) and tetraploid cells ( P = 0.0042). Comparison of melanomas positive and negative for exons 15 and 16 (MLH1) (21/36 and 15/36, respectively) showed significant differences in the percentage of aneuploid cells between 2c and 4c ( P = 0.0141) and tetraploid cells ( P = 0.0404). In summary, deletions in DNA mismatch repair proteins MSH2 and MLH1 were present both in lentigo maligna and in melanomas and correlated with DNA ploidy-related parameters in malignant melanomas.

An application of MIB antibody to the retrospective study of melanomas of oral mucosa and facial skin

A new monoclonal antibody prepared against a fragment of Ki-67 antigen MIB, from Dianova, was applied for investigation of malignant melanomas of facial skin (25 cases) and the oral cavity (25 cases), which were routinely embedded in paraffin. The values of the Ki-67 index (expressed as a percentage of positive nuclei) were correlated with TNM characteristics of tumors and patient survival. Significant correlation was found between the Ki-67 index and the level of lymph node involvement (N value), the presence of distant metastases and the time of patient survival. A positive relationship between the Ki-67 value and tumor size was also observed although it lacked statistical significance.