Biomedical subjects

R Kodet

Publications and source records attributed to R Kodet.

At least 37 records · Page 2Linked to original sources

PubMed2003

Paget's disease of the nipple: a copy number of the genes ERBB2 and CCND1 versus expression of the proteins ERBB-2 and cyclin D1.

Paget's disease of the nipple (PDN) is characterized by Paget's cells infiltrating from an underlying breast carcinoma into the epidermis of the nipple. The tumor cells were reported to overexpress ERBB-2 protein. There is no evidence, whether the overexpression is caused by amplification of the ERBB2 gene (as is the case of invasive duct carcinomas) or by other causes. Because the Paget's cells proliferate vividly we were interested also in cyclin D1 expression and in its relation to the number of CCND1 gene copies. To address these issues, we performed a study using fluorescence in situ hybridization on interphasic nuclei (I-FISH) on formalin fixed / paraffin embedded tissue sections from twelve women with PDN. We compared immunohistochemical (IHC) expression of the ERBB-2 protein and cyclin D1 with the copy number of the gene ERBB2 and CCND1, respectively (I-FISH). In all cases of our series we found overexpression of the ERBB-2 protein (3+), and in all of them there was a strong amplification of the ERBB2 gene (more than 10 signals per a nucleus, usually about 20). Keratinocytes of the epidermis had two signals of the gene. IHC and I-FISH revealed comparable findings in successive biopsies in two patients. Cyclin D1 was positive in seven cases. We found a moderate amplification of the CCND1 gene (up to 10 signals per a nucleus) in three patients only. Any correlation between the cyclin D1 overexpression anda copy number of the CCND1 gene was observed. Combined numerical changes of chromosome 17 and of chromosome 11 were found in seven women. It is concluded that in PDN, the ERBB-2 protein overexpression is caused by amplification of the ERBB2 gene. A specific immuno-therapy with trastuzumab (Herceptin) used in patients with invasive duct carcinoma may be also effective in patients with PDN.

Adult↗

PubMed2003

[Correlation of results of flow cytometry and morphologic findings in the diagnosis of malignant B-cell lymphoma].

BACKGROUND: Immunophenotyping of malignant lymphomas becomes necessary for the correct classification, for the design of therapy and for prognosis projection (WHO). Although the spectrum of classic immunohistochemical (IHC) examinations in paraffin embedded or frozen sections has recently considerably extended, IHC should preferably be combined with flow cytometry. The main advantage of flow cytometry is a synchronous application of two or more antibodies marked with various fluorochromes in one sample. The method is limited by utilizing native material only. METHODS AND RESULTS: The flow cytometry combined with histological and IHC investigations were used in diagnosis of primary non-Hodgkin's lymphomas of B-cell origin (B-NHL) and for bone marrow staging or restaging. We studied 90 patients with confirmed or suspected B-NHL and we found a good correlation in 89% of samples of primary lymphomas and in 85% of bone marrow samples when IHC and flow cytometry results were compared. The overall efficacy of the flow cytometry determination in lymphoma infiltration of the samples was 89%. CONCLUSIONS: Immunophenotyping utilizing flow cytometry contributes to diagnosis and classification of B-cell lymphomas in the significant proportion of investigated patients. In some cases it is even unnecessary to employ IHC examination of tissue sections. The method is especially suitable for determination of monoclonal populations of B-cells by detection of cell surface markers because it is more specific and sensitive than IHC. The immunophenotyping by flow cytometry as an auxiliary method and in correlation with morphological findings it can make the diagnosis of B-cell lymphomas faster and more specific.

Adult↗

PubMed2002

[Invasive ductal carcinoma of the breast: study of the number of copies of the CCND1 gene and chromosome 11 using fluorescence in situ hybridization (FISH) in comparison with expression of cyclin D1 protein and estrogen receptors (ER alpha) with immunohistochemical detection].

BACKGROUND: Overexpression of oncogenic proteins may be caused by gene amplifications. Cyclin D1 participates in regulation of the cell cycle. Relations between cyclin D1 expression and amplification of CCND1 gene encoding this protein in invasive duct breast carcinomas (IDC) are not fully elucidated. An increased interest is also focused on relations to the estrogen receptor (ER alpha). METHODS AND RESULTS: We investigated copy numbers of the CCND1 gene, expression of cyclin D1 and expression of ER alpha in a group of 60 females and 1 male with IDC. The age range varied from 33 to 89 years (median 57 years). The number of CCND1 gene copies and the number of chromosome 11 was evaluated using FISH, the expression of cyclin D1 and ER alpha was investigated by IHC. We detected a strong amplification of CCND1 gene (> 10 copies per tumor cell nuclei) in 9 patients, weak amplification (< or = copies) in 16 patients. Amplification of the CCND1 gene correlated well with the overexpression of cyclin D1. We observed the overexpression of cyclin D1 also in 13 of 36 patients without the gene amplification; therefore, the mechanism of the protein overexpression is different than that caused by the gene amplification in a proportion of patients. Amplification of the CCND1 gene was associated with a high histologic grade of IDC, whereas cases with cyclin D1 overexpression only were not. 24 of 31 patients with overexpression of cyclin D1 coexpressed ER alpha. We did not find correlation between expression/amplification of cyclin D1/CCND1 gene and the size of carcinomas and with metastases to the axillary lymph nodes. CONCLUSIONS: Amplification of the CCND1 gene is associated with overexpression of cyclin D1 in a majority of IDC. Overexpression of cyclin D1 is related to an increased expression of ER alpha. Interaction between cyclin D1 and ER alpha may explain low response to anti-estrogen therapy of some patients.

Adult↗

PubMed2002

Spindle and cuboidal renal cell carcinoma, a tumour having frequent association with nephrolithiasis: report of 11 cases including a case with hybrid conventional renal cell carcinoma/ spindle and cuboidal renal cell carcinoma components.

AIMS: We present the largest series of an unclassified subtype of renal cell carcinoma, which seems to be a distinct morphological entity and which is sometimes designated as spindle and cuboidal renal cell carcinoma. METHODS AND RESULTS: Eleven cases of spindle and cuboidal renal cell carcinoma were found among 7000 primary renal cell tumours in Pilsen's routine and consultation files. The patients were five men and six women. They ranged in age from 22 to 65 years (mean 56.8). Microscopically, the tumours were composed of two main populations of cells. First, the preponderant type of cells was formed by flattened, spindle cells with sparse cytoplasm. The second cell type was a small cuboidal cell with clear to light eosinophilic cytoplasm. Spindle-shaped cells were arranged in a fascicular pattern often reminiscent of low-grade smooth muscle tumours. Solid areas of spindle cells were also present. Small cuboidal cells formed sparse tubular structures lined by a row of single cells. In addition to all previous published cases of spindle and cuboidal renal cell carcinoma we observed an association of nephrolithiasis in our cases. It was seen in 3/11 of our patients. A previously unreported feature is the occurrence of a conventional renal cell carcinoma component in one of our cases. Seven of our patients are currently well without signs of recurrence or metastasis, one had metastasis in a regional lymph node at the time of nephrectomy, one died of an unrelated condition, and two were lost to follow-up. CONCLUSIONS: We present 11 cases of spindle and cuboidal renal cell carcinoma, which is believed to be a distinctive morphological entity. Our cases were histologically, immunohistochemically and ultrastructurally similar to the previously reported cases of spindle and cuboidal renal cell carcinoma. In contrast to all previously reported cases of spindle and cuboidal renal cell carcinoma, we observed an association with nephrolithiasis in three of our cases; moreover, one of our tumours had a conventional renal cell carcinoma component and another revealed a metastatic focus in a regional lymph node. None of our patients died of the disease. This study confirms that spindle and cuboidal renal cell carcinoma has a low malignant potential.

Adult↗

PubMed2002

[Use of computer tomography for localization of a biopsy needle for tissue collection from the orbit].

In four patients with orbital expansion the authors used computed tomography (CT) to locate the bioptic needle inserted into the orbit. In all these instances it proved possible to collect a sufficient amount of pathological tissue to assess the diagnosis. Serious side-effects were observed only in one case, where the patient developed after insertion of the needle transient bradycardia. CT was also used to locate the needle for exhausting of blood from a lymphangioma of the orbit.

Biopsy, Needle↗

PubMed2002

[Metaplastic spindle-cell (fibromatosis-like) carcinoma of the breast--report of 4 cases].

The authors report about clinico-morphological features in four cases of spindle-cell metaplastic carcinoma of the breast, closely mimicking benign soft tissue fibromatosis or nodular fasciitis. All patients were females aged 54-72 years with a palpable nodule within the breast, 20-35 mm in size. Histologically, the tumor infiltrated surrounding tissue; it consisted of spindle cells with relatively bland morphology, arranged in a fascicular, storiform or random fashion. Mitotic activity was low, focally the tumors revealed regressive changes (hyalinization, myxoid degeneration). Cytokeratin expression was proven in all four cases. In two patients the tumor metastasized and the patients died of it; this fact confirms recently published data that despite its deceptively benign appearance, fibromatosis--like carcinoma of the breast is undoubtedly a malignant process with potentially aggressive behaviour. Differential diagnosis of this unusual variant of metaplastic breast carcinoma is discussed together with related therapeutic issues.

Aged↗

PubMed2002

[Indications for treatment in invasive ductal carcinoma of the breast with Herceptin from the aspect of laboratory diagnosis--study of the ERBB-2 protein and determination of the number of copies of the ERBB2 gene. Review].

Overexpression of the ERBB-2 protein has become a target of the anti-neoplastic treatment in patients with invasive duct carcinomas of the breast with a monoclonal antibody trastuzumab (Herceptin, Genentech). From this reason the immunohistochemical (IHC) evaluation of ERBB-2 protein expression is crucial. However, there are patients in whom the IHC investigation is biased by a subjective evaluation among cases considered negative (in the range of 1+) and positive (2+), and among cases considered positive 2+ and 3+. In such cases it is advisable to complement the IHC investigation by detection of copy numbers of the ERBB2 gene with fluorescence in situ hybridization (FISH). The overexpression of the protein is caused by the gene amplification in a majority of cases. The patients, whose carcinomas show overexpression of the ERBB-2 protein and the overexpression is caused by a significant gene amplification, profit from the Herceptin therapy. In this review we focus also on the methodology of FISH in paraffin sections and tissue imprints with respect to the detection of copy numbers of chromosome 17 and the ERBB2 gene.

Antibodies, Monoclonal↗

PubMed2002

[Immunohistochemical detection of HER2 protein and its evaluation in breast carcinoma: comparative study of two methods (HercepTest DAKO versus the routine immunohistochemical approach with the DAKO polyclonal antibody) and the results from two laboratories].

Routine assessing of HER2 status is necessary for successful treatment of carcinoma of breast by Herceptin. Our results prove comparability of well standardized semiquantitative method by Hercep Test DAKO: both negative findings (73.4% in laboratory A, and 77.4% in laboratory B) and positive findings (26.6% in laboratory A, and 22.6% in laboratory B) corresponded both with each other and with the DAKO's reference data. The study also testifies to problems with interpretation of the IHC data (primary antibody c-erbB-2-oncoprotein DAKO). The problems concern particularly the processing standards, the dilution of antibodies, the application of IHC automatic machine, and probably also the detection system. It is suggested that each laboratory develops a standardized IHC protocol, in coordination with other laboratories. Thus, standardization of both processing and evaluation of the results will be achieved. As the indication for treatment by Herceptin is connected with HER2 overexpression, it is suggested to follow the algorhythm of HER2 examination presented by Nenutil at the ROCHE Satellite symposium.

Antibodies, Monoclonal↗

PubMed2001

Methylation changes in promoter and enhancer regions of the WT1 gene in Wilms' tumours.

Although the WT1 gene has been implicated in the aetiology of Wilms' tumour, mutations in WT1 are found only in minority of the tumours. DNA methylation of regulatory elements represents another possibility of modulation of gene expression. We studied methylation in the promoter and enhancer regions of the WT1 gene in 34 Wilms' tumour patients by the polymerase chain reaction on HpaII-digested DNA and by the bisulphite method. No methylation was detected in the promoter region in either tumour or normal kidney or blood DNA samples. In contrast, a HpaII site in the enhancer region was at least partially methylated in normal kidney and blood DNA samples and in about one-third of the tumours, while the majority of tumours showed no methylation. The differential methylation in the enhancer region of the WT1 gene may indicate that methylation of this element can play a role in the regulation of this gene.

Adolescent↗

PubMed2001

Idiopathic fibrosing pancreatitis presenting with obstructive jaundice in a child.

Idiopathic fibrosing pancreatitis (IFP) is a rare cause of obstructive jaundice in children. The obstruction is caused by circumferential compression of the intrapancreatic portion of the common bile duct (CBD). Various forms of biliary decompression, including surgical sphincteroplasty, choledochoduodenostomy, and Roux-en-Y choledochojejunostomy are employed as a treatment. We report a 14-year-old boy with IFP who was successfully treated by a temporary drainage of compressed CBD with the help of a stent placed by endoscopic retrograde cholangiopancreatography (ERCP). The diagnosis of IFP was confirmed histologically after open surgery. The patient remains well after 3.5 years of follow-up with no evidence of pancreatic insufficiency. This report offers an additional case of IFP causing obstructive jaundice treated by temporary internal drainage of CBD, which is safer and less invasive than biliodigestive surgical decompression.

Adolescent↗

PubMed2001

Detection of minimal bone marrow infiltration in patients with localized and metastatic Ewing sarcoma using RT-PCR.

Ewing sarcoma and related neoplasias are characterized by the presence of specific chromosomal translocations resulting in EWS/ETS gene rearrangements. Created EWS/ETS-oncogene fusion transcripts can be detected in up to 98% of ESFT and provide tumour-specific markers useful in diagnostics. Using RT-PCR for detection of this aberration we can reveal minimal amounts of tumour cells contaminating BM, blood or apheresis products. We have examined BM samples from 22 patients (21 newly diagnosed and one recurrent disease) with histologically confirmed ESFT for the presence of contaminating tumour cells in BM at the time of diagnosis. Sixteen patients presented with localized disease, six had distant metastases at the first presentation. Ewing sarcoma cells were detected in the BM of 5/16 (31%) patients with localized disease and 3/6 (50%) with clinically detectable metastases at diagnosis. BM smears prepared from the same aspirates evaluated by light microscopy were all negative, even in two patients with multiple bone disease. We have confirmed the high sensitivity of the RT-PCR assay for detection of minimal BM infiltration in localized and metastatic ESFT. We have found that more than a quarter of patients with localized ESFT have minimal BM infiltration. Although the clinical significance of the minimal disease detected at the molecular level remains unknown, RT-PCR evaluation may enable better stratification of patients into risk groups in the future.

Adolescent↗

PubMed2001

Expression of galectin-3-reactive ligands in squamous cancer and normal epithelial cells as a marker of differentiation.

The definition of biological markers for oropharynx and larynx cancer is essential to predict their clinical behavior. Since cellular glycans play an important role in biological information transfer, we have employed an endogenous lectin, galectin-3, to examine in primary squamous carcinomas, lymph node metastases, and physiological squamous epithelia whether glycans recognized by this lectin are altered in relation to the state of differentiation. The expression of galectin-3 was concomitantly evaluated by immunohistochemistry using the A1D6 monoclonal antibody. In addition, other antibodies were used for the detection of cytokeratins and desmosomal proteins (desmoplakin-1 and desmoglein). The results show the expression of galectin-3-reactive ligands in moderately/highly differentiated carcinomas only in areas exhibiting a high level of keratinization. Except for one patient out of 14, metastatic cells in lymph nodes expressed no accessible binding sites for galectin-3. No galectin-3-reactivity was detected in the basal cell layer of all studied normal epithelia (which contains the proliferating cells). The suprabasal layers were positive in epidermis and epithelium of tongue and cornea and negative in epithelium of palatine tonsil. The tumor cells expressed galectin-3 with an intensity positively correlated with tumor differentiation. The position of galectin-3-reactive sites colocalized with the two tested desmosomal proteins. However, presence of these proteins was also detected in areas of tumor and suprabasal layers of tonsil epithelium where no binding reactivity for galectin-3 was found. The present study showed that expression of galectin-3-reactive glycoligands is differentiation-dependent in normal as well as malignant squamous cells. Colocalization of galectin-3-reactive sites with desmosomal proteins (desmoplakin-1 and desmoglein) suggests an association of the galectin-3 ligand(s) with the cell surface, pointing to a potential participation of galectin-3 in mediation of intercellular contacts in these tumor types.

Antigens, Differentiation↗

PubMed2001

[Regression of hCG in various types of molar pregnancies--clinical course and prognosis].

OBJECTIVE: To evaluate spontaneous regression curves of hCG serum positivity in patients with surgically treated molar pregnancies. Comparison of complete, partial and invasive mole. The study should result in optimalisation of follow up criteria of molar pregnancies in respect to their potential malignant change. DESIGN: Retrospective comparative clinical study. SETTING: Obst. Gyn. Dpt., Oncogynecology div., 2nd Medical Faculty, FNM, Charles University Prague, Pathology Dpt., 2nd Medical Faculty, Institute of Biology and Medical Genetics. METHODS: Evaluation of spontaneous regression curves of serum hCG levels in 104 molar pregnancies. 46 patients with partial hydatiform mole, 48 patients with complete hydatiform mole, 10 patients with invasive mole. Serum hCG levels were detected by radioimunoassay (RIA) in the first period and imunochemoluminisent assay (LIA) in the second period. Regression curves of hCG positivity in particular moles were statistically evaluated by Fischer test and t-test. RESULTS: There is statistically significant difference in spontaneous regression of hCG positivity in different types of molar pregnancies. Recommended criteria for gestational trofoblastic disease (GTD) diagnosis and follow up are fully applicable in clinical practice. There is exception in partial hydatiform moles, where plateau in hCG regression does not necessarily implicate chemotherapy in patient with good compliance. CONCLUSION: Early diagnosis of GTD predominantly due to the widespread use of ultrasonography changes classical clinical features of molar pregnancies. Spontaneous regression in hCG positivity in serum is more rapid in patients with partial hydatiform mole, slower in complete hydatiform mole and invasive mole. There is no significant change in malignant potential regarding early detection and treatment.

Chorionic Gonadotropin↗

PubMed2001

[Mediastinal teratomas in children].

The objective of the work is a clinical and histopathological retrospective study of rare mediastinal teratomas in children. The authors present their experience with the treatment of eight children within the range from neonates to the age of 15 years with a histologically confirmed diagnosis of mediastinal teratoma. In two neonates the first symptom was postnatal dypnoea, teratomas in older children were diagnosed along with symptoms of acute respiratory infection. Complete resection of the tumor was performed in all patients, in neonates emergency operations were involved. Four tumors were histologically classified as mature teratomas, three as differentiated teratomas with an immature component and one as a malignant teratoma. The malignant teratoma contained mature tissues as well as tissues of embryonic carcinoma and a yolk sac tumor. Seven children with a mature or differentiated teratoma are without signs of disease 5-15 years after surgery. During the follow-up of the 15-year-old boy with the malignant teratoma the elevated alpha-fetoprotein serum level was evidence of a relapse of the disease after termination of comprehensive treatment. From the results ensues that radical extirpation is sufficient treatment for mature and differentiated teratomas. Malignant teratomas are indicated for primary resection and postoperative chemotherapy. Long-term follow up of alpha-fetoprotein and human beta-choriogonadotropin levels, clinical controls and controls by imaging methods are indicated in children with an immature or malignant component of the tumor.

Adolescent↗

PubMed2000

[High-dose chemotherapy in the treatment of children with malignant lymphoma].

BACKGROUND: Children with primary refractory or recurrent malignant lymphoma have usually poor prognosis. Less than 10% of those, who were treated with conventional-dose regimens had survived for 2 years. In an attempt to improve the outcome for these patients, we explored the role of consolidation high-dose chemotherapy with autografting. METHODS AND RESULTS: Forty-five patients with poor-prognosis lymphoma, of whom 27 were males, underwent megatherapy between January 1992 and December 1999. High-dose chemotherapy was indicated in patients with poor initial response to first-line chemotherapy (14 cases) or in the relapse (31 cases). The group consisted of 27 patients with Hodgkin's disease and 18 patients with non-Hodgkin's lymphoma. The median age was 14.7 years. The conditioning for Hodgkin's disease patients contained cyclophosphamide, etoposide and busulfan or carmustine. Patients with non-Hodgkin's lymphomas received cyclophosphamide, etoposide and busulfan or total body irradiation. Bone marrow was used as the source of haemopoietic stem cells in ten patients, peripheral blood in twenty-eight, and both sources were used in seven patients. After the median follow-up of 47 months, the final survival was 61%. Eleven patients died of the disease progression, four of the infectious complications, one at a car accident. Median time to relapse after the transplantation was 7.5 months. CONCLUSIONS: Further improvement of these results will require earlier transplantation, improved preparative regiments or early posttransplant immunotherapy.

Adolescent↗

PubMed2000

Endogenous lectins (galectins-1 and -3) as probes to detect differentiation-dependent alterations in human squamous cell carcinomas of the oropharynx and larynx.

Expression of glycan determinants for in situ binding is the prerequisite for a productive protein (lectin)-carbohydrate recognition. Labeled tissue lectins as tools are preferable to plant lectins to assess this parameter, because plant and animal lectins with identical saccharide specification can well differ in their profiles of oligosaccharide binding pattern. Due to their relevance in growth control and matrix adhesion the family of galectins (galactoside-binding metal ion independent animal lectins) is receiving increasing utilization in human biology. Employing biotinylated galectin-1 and galectin-3 we studied the expression of binding sites for these galectins in normal human squamous epithelium and human carcinomas from the oropharyngeal region and larynx in relation to the expression of LP-34+ cytokeratins by the procedure of double labeling. Tissue sites accessible for galectin-1 were located in all layers of normal epithelium and in tumor cells. In contrast, galectin-3 binding was suprabasal in the normal epithelium and in tumor cells exhibiting signs of keratinization. These results reveal differences in the localization of accessible sites for the two galectins. Relating to cell development galectin-3 appeared to display affinity to areas with increased extent of differentiation.

Antigens, Differentiation↗

PubMed2000

[Cerebrospinal fluid cytology in retinoblastoma--possibility of early detection of tumor dissemination into the CNS].

Retinoblastoma is a malignant tumourous which occurs in childhood. The most important factors which influence possible cure of patients with a retinoblastoma are early detection of the disease, correct diagnosis and adequate therapy. In the submitted case-record the authors provide evidence of the importance of cytological examination of cerebrospinal fluid which is important in retinoblastoma in particular for the follow-up of treatment. Cytological examination of cerebrospinal fluid in this type of tumour is very reliable and can reveal infiltration of tumour cells into the CNS much sooner than imaging methods.

Cerebrospinal Fluid↗

PubMed2000

[Abdominal lymphangiomas in childhood].

The authors evaluate their experience with the diagnosis and treatment of abdominal lymphangiomas during the period of 1995-1999. During the above period 6 girls and 4 boys with abdominal lymphangiomas were operated. The mean age at the time of operation was 5 years and 8 months. Ultrasonographic examination was made in all 10 patients, CT examination in 8. The surgical finding was lymphangioma of the mesenterium 4x, of the omentum 2x, of the adrenals 2x of the retroperitoneum 1x and intestinal lymphangiomatosis 1x. Macroscopically complete extirpation of the lymphangioma was possible in 9 patients and called for resection of the gut 3x and for adrenalectomy 2x. In one patient subtotal extirpation was performed. Postoperative follow-up did not reveal a relapse in any of the children. Abdominal cystic lymphangiomas are rare benign malformations of the lymphatic system. For preoperative differential diagnosis in typical cases USG is sufficient. The therapeutic method is complete surgical extirpation which is associated with a low incidence of relapses.

Abdomen↗