Jacek Gawrychowski

Prognosis of the surgical treatment of patients with non-small cell lung cancer (NSCLC) – relation to DNA ploidy

OBJECTIVE The aim of this study was to evaluate prognostic importance of cell ploidy and proliferation activity in non-small cell lung cancers. Survivals were compared according to the following factors: sex, age, histology, grading, DNA ploidy, tumour size, T factor, N factor and operative procedure. METHODS In a group of 191 patients in whom cytofluorometric examinations had been performed on archival tumour specimens, postoperative recurrences were observed. RESULTS Postoperative recurrence was observed in 64 (64.6%) of 99 patients with aneuploid tumours and in 35 (38.0%) of 92 with diploid tumours (P<0.001). Overall survival (OS) rates for the group of 92 patients operated for diploid non-small cell lung cancer (NSCLC) at 5 and 10 years were 62 and 51.1%, whereas of other 99, operated for aneuploid tumours 33.3 and 25.9%, respectively (P<0.001). In the former group of patients disease-free survival (DFS) rates at 5 and 10 years were 58.7 and 51.4% but in the latter 29.3 and 26%, respectively (P=0.00014). Significant differences dependent on cell ploidy were also observed in OS and DFS rates of patients operated respectively for SCLC (P=0.0029; P=0.00318) and adenocarcinoma (AC; P=0.0241; P=0.02109). In general, the mean percentage of S-phase cells in non-small cell lung cancers was 14.0% (SD=13.1) in patients who survived 5 years, and 22.4% (SD=15.7) in those who had a recurrence or died (P<0.001). CONCLUSIONS In our opinion the most important finding of our work is that determination of cell ploidy in NSCLC provides a valuable supplement to the TNM stage when evaluating late results of the surgical treatment. However, the paper demonstrates that aneuploidy, although unfavourable, is not an independent prognostic factor in the group of patients with NSCLC and in the subgroups - both with squamous cell carcinoma and adenocarcinoma. Our results show also that the percentage of S-phase cells is an independent, unfavourable prognostic factor in patients treated surgically for non-small cell lung cancer and in the subgroup with squamous cell lung carcinoma.

Molecular profiling in primary hyperparathyroidism

Primary hyperparathyroidism (HPT) is one of the most common endocrine disorders, defined by hypersecretion of parathormone. Primary HPT can be caused by adenoma, hyperplasia, and carcinoma. A great amount of mechanisms contribute to the pathogenesis of this disease, such as genetic predispositions because of the germline‐inactivating mutations in the multiple endocrine neoplasia type 1 (MEN1) and HRPT2 tumor suppressor genes. Somatic mutations in these genes were found also in sporadic parathyroid neoplasias. Cell cycle regulators, growth factors, apoptosis‐inducing ligands, death receptors, and other transmitter substances have also been implicated in the etiology of primary HPT. Parathyroid carcinoma is often misdiagnosed as parathyroid adenoma and long‐term survival is conditioned by the extent of the primary surgical resection, therefore, of great interest is the discovery of definitive diagnostic markers for carcinoma. This article presents current state of knowledge of the molecular pathogenesis of primary HPT.

Imaging diagnostics for primary hyperparathyroidism

Primary hyperparathyroidism (PHP) is a benign condition characterised by malignant potential. Even in specialist wards, 5-10% of operations for PHP are unsuccessful. The main reasons seem to be ectopy of the parathyroid gland, numerous adenomas, multiglandular parathyroid hyperplasia, and intrathyroid location of the parathyroid. The last three decades have witnessed a rapid progression in imaging diagnostics.

Oxidative stress and angiogenesis in primary hyperparathyroidism

SummaryBackgroundThe inappropriate elevation of parathormone (PTH), which regulates the process of angiogenesis in parathyroid tissue, causes the changes of activity of enzymes responsible for the removal of free radicals. Parathyroidectomy (PTX) in patients with primary hyperparathyroidism (PHPT) lowers the level of PTH and leads to the reduction of risk of cardiovascular and all-cause mortality by normalization of the antioxidant status. Therefore, the aims of the study were to assess the activity of antioxidant enzymes and free radical reaction products in patients after parathyroidectomy, and to evaluate the correlation between the systemic oxidative stress and angiogenic parameters.Materials and methodsPatients with PHPT treated surgically were enrolled into the study. Total antioxidant capacity (TAC), total oxidative status (TOS), oxidative stress index (OSI), superoxide dismutase (SOD), ceruloplasmin (CER), lipid hydroperoxides (LHP) and malondialdehyde (MDA) were measured before and after parathyroidectomy. The immunohistological expression of angiogenic factors in parathyroid specimens was assessed by the BrightVision method from ImmunoLogic using murine monoclonal anti-human: anti-VEGF, anti-CD31 and anti-CD106 antibodies.ResultsThe significant increase of TAC, CER, reduction of TOS, MDA, SOD, especially for cytoplasmic form, and significant decrease of OSI, LHP were observed after PTX. There was no significant correlation between changes of oxidative stress markers and angiogenic parameters: VEGF, CD-31, CD-106 in parathyroid tissue. The correlation level was low and medium.ConclusionsParathyroidectomy causes down-regulation of lipid peroxidation processes and leads to reduction of oxidative stress in patients with PHPT. The decrease in the OSI is the results of down-regulation of oxidative stress in the postoperative period. The change of the antioxidant status has no impact on angiogenesis processes in parathyroid tissue.

Expression of TRAIL and Fas in Primary Hyperparathyroidism

ABSTRACT Aim: Differentiating between parathyroid lesions is still difficult and ambiguous. In cases of primary hyperparathyroidism, appropriate and prompt diagnosis is of great importance for effective treatment and follow-up. A great amount of mechanisms contribute to the pathogenesis of primary hyperparathyroidism, such as disturbance in balance between pro- and anti-apoptotic factors. Therefore, we examined whether immunohistochemical expression of apoptotic factors, TNF-related apoptosis-inducing ligand (TRAIL) and Fas, could have clinical utility as a marker of proliferative lesions of parathyroid gland. Materials and methods: Parathyroid specimens of 58 consecutive patients who had undertaken surgery due to primary hyperparathyroidism were incubated with purified mouse monoclonal antihuman antibodies: anti-TRAIL and anti-Fas. Staining was considered positive when at least 5% of the cells showed immunoreactivity. Results: The percentage of cells which were positively stained for TRAIL in parathyroid hyperplasia was 9.65%, in parathyroid adenoma 8.31%, and in normal controls 2.24%. Immunoreactivity for TRAIL was detected in 91.89% of parathyroid hyperplasias, 85.71% of parathyroid adenomas, and none in healthy glands. The percentage of cells with a positive reaction to Fas in parathyroid hyperplasia was 8.92%, in parathyroid adenoma 8.09%, and in normal tissue 1.9%. The expression of Fas was found in 94.59% of parathyroid hyperplasias, 90.48% of parathyroid adenomas, and none in healthy glands. Conclusions: In our study, hyperplasias demonstrated the highest expression of TRAIL and Fas, whereas in adenomas it was increased compared to normal tissue, but lower than in hyperplasias. These factors could be an additive tool in the differential diagnosis of parathyroid lesions.

Non-Recurrent Laryngeal Nerve.

UNLABELLED The aim of the study was to assess the frequency of non-recurrent laryngeal nerves (Non-RLN). MATERIAL AND METHODS A total of 6110 patients were operated in our hospital between 1 January 2005 and 31 December 2013 for various goiters (5429) and various types of hyperparathyroidism (618). Laryngeal nerve was exposed during operation in 1700 patients from superior aperture of the chest to superior aperture of the larynx. Identification process of RLN was started with dissecting inferior thyroid artery (ITA) and its junction with the nerve. Then main trunk of the nerve was exposed backwards till the region of superior aperture of the chest together with the end portion till the nerve outlet to the larynx. RESULTS In the group of 1700 patients, RLN was exposed bilaterally in 1400 (82.4%) and unilaterally in 300 (17.6%). In the group of 3100 dissected RLNs the course of RLN was observed on the right side in 1710 patients and on the left in 1390. Irreversible nature RLN was shown in four cases (0.1%) - four women (02%) aged 42-55 (mean 49.3) - three operated for non-toxic nodular goiter and one for primary hyperparathyroidism. Each time the Non-RLN was seen on the right side. The other patients manifested recurrent character RLN. Moreover, interstitial course of RLN was found on the left side in one man. CONCLUSION Non recurrent laryngeal nerve is a rare anatomical variation, occurring more frequently on the right side. Surgeon during surgery of the thyroid and parathyroid glands should be aware of its existence to avoid damage.

Tissue hemostasis and chronic inflammation in colon biopsies of patients with inflammatory bowel disease.

Inflammatory bowel disease (IBD) is characterized by a chronic inflammation accompanied by procoagulation settings. However, tissue hemostasis in IBD patients was only incidentally reported. Accordingly, the current study characterizes changes in tissue hemostasis components in a colon inflammatory setting. Serial cryostat sections of endoscopic mucosal biopsy specimens taken from 26 consecutive IBD patients diagnosed de novo and normal colon resection specimens taken from 6 patients were immunohistochemically stained with monoclonal anti-human tissue factor (TF), tissue factor pathway inhibitor (TFPI), thrombomodulin (TM), as well as CD3 and CD68 positive cells. The hemostatic components studied differed significantly from the control subjects. Up-regulation predominated in the case of TF while down-regulation was mainly found in TM and TFPI in IBD. In the control sections, TF was observed in a few fibroblast-shaped cells in the lamina propria, while in the majority of IBD sections, TF positively stained small microvessels, infiltrating mononuclear cells and fibroblast-shaped cells tightly surrounding the colon crypts. Thrombomodulin intensively stained the endothelium of the small capillary vessels in the control, whereas such staining mainly accompanied infiltrating mononuclear cells of the IBD subjects. Tissue factor pathway inhibitor positively stained the endothelium of the small capillary vessels in the control group, whereas in the IBD group endothelial cells presented only weak TFPI staining. The mean number of CD3-positive lymphocytes in IBD was 23.3 ± 14.3, but the mean number of CD68-positive cells was 114.5 ± 55.8. In the control sections, it was 4.1 ± 2.4 and 39.6 ± 17.9, respectively. There was no relationship between CD3 and CD68 (+) cells and the hemostasis markers studied. The results of the current study indicate a shift of tissue hemostasis toward the procoagulant state irrespective of the severity of inflammatory infiltration. In addition, TF distribution in the colon sections of IBD patients may indicate a role in the restoration of the barrier function in injured intestinal mucosa.

A follow-up study of patients with MEN syndromes — five case reports

THE AIM OF THE STUDY Evaluation of the results of surgical treatment in patients with hyperparathyroidism related to MEN syndrome MATERIALS AND METHODS: The group of three women and two men aged between 17 and 72 years ( av. 38.6) with MEN- related hyperparathyroidism within 419 patients were operated between 2010-2016. MEN1 syndrome was diagnosed in four patients and MEN2 only once. Pre- and postoperative results of PTH levels and serum calcium values were compared. RESULTS The mean preoperative serum PTH level was 215,56 pg/ml, whereas calcium concentration was 1.40 mmol/l. Three Patients with MEN1 syndrome had three and two thirds of parathyroid gland taken, and one had a single parathyroid gland excised with biopsy of the rest glands performed. A patient with MEN2 syndrome had one parathyroid gland excised. Postoperatively, we reported decreased secretion of both PTH (a mean serum PTH concentration 6.72 pg/ml), and serum calcium concentration 1.11 mmol/l. One patient (25%) with MEN 1 developed recurrent hyperparathyroidism after five years from the surgery. The patient was reoperated. The rest of the parathyroid gland was removed with an implantation into separated muscle pockets in the anterior forearm muscles. CONCLUSIONS 1. MEN syndrome is a rare cause of primary hyperparathyroidism. 2. Subtotal parathyroidectomy helps to bring back a normal calcium metabolism. 3. Recurrence of hyperparathyroidism in patients with MEN syndrome requires total parathyroidectomy with an autotransplantation into the anterior forearm muscles.

Telomerase activity in non-small cell lung cancer.

Introduction High telomerase activity has been detected in the majority of malignant neoplasms including lung cancer. The purpose of the study was to attempt to use telomerase activity as a prognostic factor in patients with non-small cell lung cancer (NSCLC). Material and methods Telomerase activity was analyzed in 47 tissue specimens taken from patients with NSCLC. The control group consisted of 30 specimens of non-cancerous lung parenchyma. Telomerase activity was measured by means of the telomeric repeat amplification protocol (TRAP). Results Telomerase activity in the neoplastic tissue was significantly higher than in the lung parenchyma that was free from neoplastic infiltration. There was no significant association between telomerase activity and age, gender, tobacco smoking, histological type of the tumor, or staging (pTNM). No association was found between the level of telomerase activity in NSCLC specimens and the two-year survival rate of patients (p = 0.326). A higher level of telomerase activity in poorly differentiated tumors (G3) as compared to moderately differentiated tumors (G2) was detected (p = 0.008). A positive association was identified between telomerase activity in pulmonary parenchyma free from tumor infiltration and the presence of leukocyte infiltration (p = 0.0001). Conclusions No association was found between the level of telomerase activity in NSCLC specimens and the two-year survival rate of patients. The study has revealed a positive association between telomerase activity and the grade of differentiation (G) in NSCLC.

Assessment of operative treatment of patients with tertiary hyperparathyroidism after kidney transplantation

INTRODUCTION The purpose of the study was to assess the results of operative treatment of patients with tertiary hyperparathyroidism (tHPT) after kidney transplantation. MATERIAL AND METHODS The study included 30 patients in whom kidney transplantation was performed between 2006 and 2013, and in whom parathyroidectomy had to be performed at a later time because of tHPT. There were 17 (56.7%) women and 13 (43,3%) men in the group, aged 18-64, mean 46.1 years. In order to locate the lesion before the operation, all patients had to undergo USG, and 14 had scintigraphy MIBI in addition. Serum levels of PTH, ionised calcium, and creatinine were determined together with glomerular filtration rate (GFR). The results of control tests were compared with those performed one day before parathyroidectomy (PTX) and three days after the operation. RESULTS Among 30 patients, 19 (63.3%) underwent total resection of three parathyroid glands and 3/4 of the fourth. Two parathyroid glands were resected in eight (26.7%) patients, and one in the remaining three (10%) patients. Histopathological examination showed one parathyroid adenoma in six (20%) patients, and one parathyroid adenoma and hyperplasia of the remaining glands in one (3.3%) patient. Five (16.7%) patients had hyperplasia of two parathyroid glands whereas no changes were observed in two patients. On the other hand, hyperplasia of all glands was noted in 18 (60%) patients. Serum PTH level was significantly lower compared to the level before operation (p < 0.001), being 5.5-58.5 pg/mL, on day 3 postoperatively. Differences in the levels of serum-ionised calcium were also significant (p < 0.0001) after eight months. CONCLUSIONS Surgical resection of parathyroid glands is a management of choice in patients after kidney transplantation accompanied by hypercalcaemia lasting longer than one year. Resection of 3 3/4 parathyroid glands because of hyperplasia in patients with hyperparathyroidism after kidney transplantation enables restoration of normal calcium metabolism. Moreover, resection of 3 3/4 parathyroid glands can allow avoidance of autotransplantation, which is necessary in cases of total resection of parathyroid glands.