Novel RET mutations in macedonian patients with medullary thyroid carcinoma: genotype-phenotype correlations.

Medullary thyroid carcinomas (MTCs) are rare neoplasms comprising 2-10% of all thyroid malignnancies. More than 75% are sporadic tumors and the remainder is familial and MEN2 related. Both sporadic and syndromic MTCs frequently show mutations in the RET proto-oncogene. It has been noted that some MTC cases present an indolent, and some an aggressive clinical course. Ki-67 expression is generally low, with documented exceptions, whereas high expression of Bcl-2 has been reported in majority of the cases. Some studies have shown that Ki-67 and Bcl-2 expressions have prognostic value, as well as RET mutational status. We analyzed 20 unrelated MTC cases for Ki-67, Bcl-2 expression and RET mutations and tested their intercorrelations, correlations to the morphologic features and stage of the tumors, as well as their influence on survival. In 13 of the 20 analyzed cases we found 23 sequence changes distributed in exons 8, 10-13 and 16. There were 11 different missense mutations, single nucleotide deletion with frameshift, and 8 different synonymous mutations. Only 4 of the sequence changes have been previously published. Twelve patients (60%) had tumors expressing one or more missense mutations or single nucleotide deletion and 7 of them (35%) had at least one damaging or possibly damaging RET mutation. Most of the tumors had low Ki-67 expression (mean 6.48% of cells) and high Bcl-2 expression (mean 68.3%). Significantly better survival was observed in cases with low Ki-67 (< 6.5%; p < 0.05), high Bcl-2 expression (> 68.3%; p < 0.01) and younger age at diagnosis (< 51 years; p < 0.05).

Synchronous occurrence of ileal stromal tumor (GIST) and colonic adenocarcinoma: a case report.

INTRODUCTION: Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors of the digestive tract. There is an increasing number of literature reports on synchronous occurrence of gastrointestinal stromal tumors and another malignancy of distinct etiology and evolution. The most reported cases include gastric synchronous occurrence of gastrointestinal stromal tumors and adenocarcinoma and gastric gastrointestinal stromal tumors and colonic adenocarcinoma. CASE REPORT: We present a case of a 77-old female, with synchronous cecal moderately differentiated adenocarcinoma in Stage IIA according to the TNM classification and ileal spindle cell type GIST with low malignant potential, positive for c-Kit, CD34, vimentin, Actin, and negative for S100. CONCLUSION: The synchronous occurrence of small bowel gastrointestinal stromal tumors and other primary gastrointestinal malignancies has been rarely reported. There is a need of further investigations to identify the relationship between gastrointestinal stromal tumors and colorectal cancers.

Digital quantitative immunofluorescent detection of p53 protein in urinary bladder cancer tissue samples.

Bladder cancer is a significant health problem and is the fourth most common malignancy in the Western world. Histologically, transitional cell carcinoma (TCC) is the most common subtype and represents nearly 90% of all bladder cancers. Mutations of the tumour suppressor gene p53 are the most frequent genetic alteration found in human cancers, including urinary bladder carcinoma. Numerous studies have attempted to define p53 as a molecular marker with clinicopathological and predictive values. In this study we have investigated tissue samples of histopathologically confirmed TCC of the urinary bladder from 70 patients. Normal urinary bladder mucosa obtained from 40 patients with nonmalignant diseases were used as a negative control group. Immunofluorescence detection was performed using double-sandwich antibody technique, the microscope images were captured by digital camera and analyzed by ImageJ software. Corrected fluorescent intensity values corresponding to average malignant cells' nuclear p53 staining were calculated from each patient sample. We found that the mean corrected fluorescence intensity values of TCC samples grade I and II did not differ statistically significantly. On the contrary, those values were significant between the TCC grades II and III, as well as between grade I and III. In addition, a high correlation of p53 fluorescence intensity values from superficial TCC samples with the invasive TCC samples was found. However, the data showed no predictive value of p53 nuclear accumulation in TCC samples in terms of recidive, metastasis or cancer-related death, at least within the 2-years follow-up period. Given the relatively small size and heterogeneity of our patient groups, however, further studies are needed to demonstrate the clinicopathological value of digital quantitative immunofluorescent detection of nuclear p53 in TCC samples.

The occurrence of acute subdural haematoma and diffuse axonal injury as two typical acceleration injuries.

Closed head injuries have already been classified into contact injuries and acceleration-deceleration injuries. Two typical acceleration-deceleration injuries and at the same time, the two worst head injuries are acute subdural haematoma (ASDH) and diffuse axonal injury (DAI), and that is where they got their medico-legal importance. Using experiments, it has been shown that acceleration with an impact time of more than 20-25 min (which occurs in traffic accidents in real life) causes DAI, whereas an impact time of 5-10 min is more likely to produce acute subdural haematoma. The aim of this research is to show that not all, but some types of traffic accidents are more typical for the occurrence of DAI, as well as that the ASDH is not a common feature for all types of fall. The analysis conveyed covered 80 cases of closed head injuries (traffic accidents, falls and assaults) where a complete forensic medical autopsy has been undertaken, followed by a complete forensic-neuropathological examination. For the purpose of diagnosing DAI, immunohistochemistry using antibody against ß-amyloid precursor protein has been involved. Results show that ASDH is more likely to occur in cases of simple fall, assaults and cyclists and DAI is more typical for vehicular traffic accidents and cases of falling from a considerable height. The paper also comprises discussion about some open questions regarding the diagnosis of DAI in the medico-legal practice.

Morphology of renal interstitial fibroblasts.

INTRODUCTION: Renal fibrogenesis is a process common to all progressive kidney diseases. The main executive cell in this process is the fibroblast, by secreting and remodelling the extracellular matrix. The number of fibroblasts is minor in a healthy kidney interstitium, but it increases during the process of fibrosis. Their morphology and immunophenotype vary due to different intrinsic and extrinsic factors which makes their identification and visualization, as well as determination of their origin, very difficult. MATERIAL AND METHODS: We performed morphological and immunohistochemical analyses on kidney biopsies with interstitial fibrosis, using the following antibodies: Vimentin, α-SMA, S100A4, Cadherin 9 and CD34. We also did light-microscopy analyses of semithin sections of tissue embedded in epoxy resin and stained with Toluidine blue. RESULTS: Our observations show that different cells in the fibroblastic population show positivity for different markers, thus contributing to the theory that there are different subpopulations of fibroblasts, with different origins, that take part in renal fibrogenesis.

Dilemmas concerning the diffuse axonal injury as a clinicopathological entity in forensic medical practice.

Dilemmas and discussions concerning the diffuse axonal injury (DAI) and still existing in forensic medical practice are as it follows: 1. Whether the occurrence of DAI can indicate the type of traumatic event that has caused the head trauma, 2. Whether the presence of axonal damage in cases of hypoxia, ischaemia and other pathological conditions casts a shadow on the post-mortem pathological diagnosis of DAI and totally negates it, or there are certain clues in the findings that can point to the aetiology of the axonal damage. This paper discusses our findings based on neuropathological examination of 60 forensic cases of closed head injury. The neuropathological examination included: a macroscopic examination of the coronal sections and a microscopic examination involving an immunohistochemical method with antibody against ß-amyloid precursor protein. Our findings indicate that DAI, as a clinicopathological entity, is undoubtedly an acceleration-deceleration injury, predominant in road traffic accidents as it is classically outlined, and cases of falling from a considerable height. Our findings point to a certain difference between the features of traumatic and ischaemic axonal damage. In this paper we also investigate the correlation between pathological grades of DAI and the impairment of the brain function before death.

The importance of the detail forensic-neuropathological examination in the determination of the diffuse brain injuries.

UNLABELLED: According to the contemporary classification, traumatic brain damage is divided on focal and diffuse brain injuries, and primary and secondary brain damage. The aim of this paper is to emphasize the necessity of the forensic-neuropathological examination in the determination of the diffuse brain injuries. In those injuries frequently neither the most sophisticated clinical-investigation techniques like CT and MRI, nor the routine post-mortem forensic pathological examination, give any results with discovering an intracranial mass lesion, despite the fact that patients had manifested a serious brain failure. In a series of 80 cases with closed head injuries where forensic-neuropathological examination has been undertaken (examination of a fixed brain tissue and immunohistochemistry using monoclonal antibodies against ß-amyloid precursor protein), the occurrence of the diffuse brain injuries in the absence of any other massive intracranial lesion has been established in 14 (17,7%) of the cases. Hence, forensic-neuropathological examination has been the only way to establish the diagnosis of the brain injury that caused a serious brain failure and in most of them occurred as a concrete cause of death. This method has already been affirmed in the forensic medicine science and has been implemented in a Recommendation No 99 of the Council of Europe where medico-legal autopsy rules are given, thus, establishing it as an unavoidable part of the daily forensic medicine practice. KEYWORDS: diffuse axonal injury - diffuse vascular injury - closed head injuries - traumatic brain damage - diffuse brain damage.

Variations of the histomorphological characteristics of human skin of different body regions in subjects of different age.

The aim of this paper was to create a reference model for the qualitative and quantitative characteristics of healthy human skin in different body regions and different life periods. For this purpose we have taken skin biopsy specimens from 15 different body regions: capillitium, forehead, cheeks, anterior neck, thorax, axilla, abdomen, back, gluteus, anterior arm, anterior forearm, palm, anterior leg, anterior lower leg and sole. The biopsies were histologically elaborated according to a standard paraffin technique, and the obtained histological slides were qualitatively and quantitatively analysed with the use of a computer system for image processing and analysis (Lucia M, Version 3, System for Image Processing and Analysis). The examinees were divided by age into five groups: from full-term infants up to the age of 1 year; from the age of 2 up to the age of 12 years; from the age of 13 up to the age of 22; from the age of 23 up to the age of 55; from the age of 56 up to the age of 73. In each exemplar were determined: the total skin thickness in each region at each age group, total thickness of the epidermis, total thickness of the corium, thickness of the papillary and reticular layers of the corium. In this period the thickening is about 4-4.5 times. The growth of the thickness comes as a result of the growth of the thickness of the reticular corium, values of which grow by 4-5 times. The height of the epidermis in newborns shows higher values than the second group (childhood). In the third and fourth group the values of the epidermis are from 1.5 to 2.5 times higher on those parts of the body which are uncovered and exposed to externalities. The essence of the changes that happen to the skin is structural final formation, which is turbulent and targeted in youth (in order to harmonize structural and functional abilities of the human organism) and in mature age to synchronize the function of the skin with the other systems of the organism.

Evaluation of telomerase activity in patients with chronic B lymphocytic leukemia versus age matched controls. Correlation between telomerase activity and bone marrow infiltration.

Telomerase is a ribonucleoproteic enzyme associated with cellular immortality and malignancy. This enzyme, besides the catalytic subunit bearing reverse transctiptase activity, contains an RNA template complementary to TTAGGG telomeric repeats, thus permitting de novo synthesis of telomeric DNA onto chromosomal telomeric ends. Increased telomerase activity has been reported in Chronic Lymphocytic Leukemia (CLL) by many authors. In order to investigate the telomerase activity in patients with CLL and its correlation to commonly used morphologic prognostic markers, 38 frozen blood lymphocyte samples from patients with CLL and 47 age-matched controls were investigated for telomerase activity using the Telomerase PCR ELISA-plus kit from Roche. Trepanobiopsies from the same patients were analysed for the type of bone marrow infiltration as well. Analysis showed highly variable Relative Telomerase Activity (RTA) in B-CLL patients, ranging from comparable or even lower than the mean RTA of controls (in Binet A stage patients) to manifold increase in the majority of patients with advanced stage disease. The sex and age of the patients showed no influence on RTA in CLL patients, in contrast to the control group, where the age influenced telomerase activity. We found a positive correlation between the RTA and disease stages (Binet), as well as between RTA and the type of BM infiltration.

Imaging diagnosis of musculoskeletal hydatid disease.

Hydatid disease has a worldwide distribution and causes health problems in endemic countries. Musculoskeletal hydatid disease may be a difficult differential diagnosis. The aim of this retrospective study is to present some different imaging features of musculoskeletal hydatid disease. We evaluated imaging findings of 15 patients with musculoskeletal hydatid disease over a 15-year period. Ten (67%) were men and 5 (33%) women, ages ranging between 12 and 69 years. All underwent radiographic evaluation. CT was done in 9 patients and MRI in 3 patients with soft tissue involvement. All underwent surgery, followed by histological examination. Bone involvement occurred in 11 patients and soft-tissue involvement in 4 patients respectively. Bone lesions in the spine were found in 4 (36%), in the pelvis 2 (18%), in the proximal femur 2 (18%), in the tibia 2 (18%) and 1 (9%) in the first metatarsal. On plain film we identified three different patterns of presentation of bone HD, defined as "typical" osteolytic lesion (33%), "tumour-like" features (41.6%) and lesions resembling infection (25%). Soft-tissue hydatidosis was localized on the distal part of the thigh, paravertebral and deltoid muscle. MR imaging showed a cystic mass, containing multiple vesicles, and in one of them the pathognomonic for hydatid disease, the "water lily" sign, was present. We present three cases of primary musculoskeletal hydatid disease localized on metatarsal bone, proximal femur and deltoid muscle. Musculoskeletal hydatid disease, presenting with a variety of patterns, may resemble a tumour or an inflammatory process. Because of the rare presentation of the disease it should be kept in mind in the diagnostic work-up of musculoskeletal tumours.

High-grade surface osteosarcoma - chondroblastic type.

We present the case of an 8-year-old boy with pain in the distal part of the right thigh that occurred after a ball injury, without haematoma or oedema. The plain film showed exostosis of the distal part of the right femur. In another institution the "exostosis-like" formation was surgically removed and the histological finding was an aggressive type of chondroblastoma. A few months later, on the site of the surgical intervention, a significantly enlarged painful solid mass appeared. Radiography showed a large dense mass at the level of distal femoral metadiaphysis with tumour matrix mineralization, and Codman's sign. Computed tomography showed a large soft tissue mass, which arose from the surface of the underlying cortical bone and signs of high-grade malignancy. CT is a valuable method in predicting high-grade malignancy. Arteriography was also done. After operative treatment - seq. Campanacci, histology showed high-grade surface osteosarcoma of chondroblastic type. Pre- and post-operative chemotherapy was carried out. The boy died 2 years after diagnosis because of recurrent disease. High-grade surface osteosarcoma has the worst prognosis, compared with other surface osteosarcomas. Therefore, timely diagnosis is indispensable.