Professor Witold Nowicki - a greatly spirited pathologist.

This paper presents a complete overview of the scientific, professional and social activity of a great Polish pathologist, Witold Nowicki (1878-1941), from mainly Polish-written, original sources with a major impact on mostly his own publications. The biographical commemoration of this eminent professor is not only due to the fact that he provided a profound microscopic characterization of pneumatosis cystoides in 1909 and 1924. Nowicki greatly influenced the development of anatomical pathology in Poland, having authored over 82 publications, with special reference to tuberculosis, lung cancer, sarcomatous carcinomas, scleroma and others. However, the first of all his merits for the readership of Polish pathologists was his textbook titled Anatomical Pathology, which was a basic pathology manual in pre-war Poland. Witold Nowicki - as the head of the academic pathological anatomy department and former dean of the medical faculty - was shot with other professors by Nazi Germans in the Wuleckie hills in Lvov during World War Two. Professor Nowicki was described as being "small in size but great in spirit" by one of his associates, and remains an outstanding example of a meticulous pathologist, a patient tutor and a great social activist to follow.

Coexistence of mature cystic teratoma and adenocarcinoma in situ within atypical proliferative mucinous tumour of ovary--a case report of 35-year-old woman.

Combined ovarian tumors are found in common pathologic practice due to amazing potential of ovarian tissue to copy almost every tissue of human body and imitate many neoplasms of various other organs in a very flexible way. A multicystic tumor is presented in this case report of 35-year-old woman. It consisted of a cyst with sebum and hair and cavities with papillomatous projections and mucus. The ovarian tumor was diagnosed a mature cystic teratoma presenting mainly as dermoid cyst and mucinous adenocarcinoma in situ, arising within atypical proliferative mucinous tumor. This report demonstrates how histoformative properties are reflected in ovarian tumorigenesis. Such a stunning histoformativity makes ovaries the possible site of primary origin for malignant tumors that mimic extra ovarian differentiation. In the authors' point of view, the diagnosis of primary ovarian mucinous tumor within cystic teratoma is firm, whenever simultaneous extraovarian involvement by mucinous neoplasm is excluded.

Acardius acephalus fetus--report of distinctive anatomical anomalies with regards to pathophysiology of TRAP sequence.

Acardiac fetuses are consequences of twin reversed arterial perfusion (TRAP). Here the authors present a case of 40-year-old gravida IX who gave birth to a healthy, 2,900 g female child by a cesarean section. Additionally amorphic 1,020 g maldeveloped fetus was removed. There was a diamnion monochorionic type of twin placenta with incorrect single umbilical arteries (SUA) both in umbilical cord of healthy fetus and in atrophic second umbilical cord. A malformed fetus developed a rather well formed lower leg with four digital foot and oval shape amorphous body mass with omphalocele and eventration of the intestines. X-ray picture showed well visible metatarsal and femur bone and anatomically undefined bones cluster in the central part. A cavity of fetal body contained intestines--the only one well-formed organ, nests of heterotopic pilosebaceous residues, remnants of adrenal glands, well-formed ganglia, and nests of neural tissue covered by neuroepithelium.

The Bak protein expression in germinal centers of hypertrophied adenoids in children.

The aim of this study was to evaluate pro-apoptotic Bak expression in the germinal centers of adenoid in children on the assumption of the potential usefulness of Bak as adenoid function marker. The study involved 95 children undergoing adenoidectomy; divided into three age groups: aged up to 5 years (25 children), 5-10 years (54 children) and over 10 years (16 children). The analyzed material was adenoids removed on the ground of hypertrophy. Immunohistochemical analyses were carried out using goat polyclonal Bak antibodies (DAKO) directed against human Bak protein. The presence of Bak positive lymphocytes within germinal centers and Bak immunostaining were scored. The immunohistochemical staining showed the Bak positive lymphocytes mainly within the germinal centers of the lymphoid follicles. The Bak reactivity was also present in hyperplastic lymphoid tissue within the subepithelial B lymphocytes. We have not found statistically significant correlation between Bak expression and clinical status and change in Bak expression level according to age. The apoptotic presence within the germinal centers are the manifestation of which is Bak expression and its lack in the mantle zone, what we confirmed in our former study by describing Bcl-2 expression, seems to be a proper B cells maturation marker within lymphoid follicles. Our finding shows that these processes are not influenced by age and supports our thesis that adenoid involution is rather the effect of changes in the number of lymphoid follicles that changes in them.

Serum erythropoietin and angiogenetic factors in human colorectal cancer.

AIMS AND BACKGROUND: Erythropoietin, VEGF, VE-cadherin are involved in angiogenesis. Besides that erythropoietin stimulates erythropoiesis and increases haemoglobin and hematocrit levels as well. Moreover, erythropoietin could directly stimulate colorectal cancer cell growth due to the presence of both erythropoietin receptor and erythropoietin production in malignant cells of this neoplasm. Therefore we aimed at measurement and comparison of serum erythropoietin with VEGF, VE-cadherin levels, blood haemoglobin and hematocrit in colorectal cancer patients of different clinicopathological profiles. METHODS: We applied ELISA kits to evaluate preoperative serum levels of endogenous erythropoietin, VEGF and VE-cadherin in samples from 92 colorectal cancer patients and control group of 16 healthy volunteers. RESULTS: Endogenous erythropoietin was significantly elevated in preoperative sera in colorectal cancer patients (p = 0.013) compared with healthy volunteers, however, erythropoietin levels were not significantly higher with the advancement of colorectal cancer. There were significantly higher levels of erythropoietin in the group of anaemic men in comparison to men with normal haemoglobin levels (p < 0.0001). VEGF and VE-cadherin did not correlate with erythropoietin. Erythropoietin levels negatively correlated with haemoglobin and hematocrit levels in all cancer patients; particularly in node positive cancers (N+), moderately differentiated tumours (G2) and deeply invading neoplasms (pT3+pT4). CONCLUSIONS: Erythropoietin levels increase in colorectal cancer but circulating erythropoietin does not associate with progression of the disease. Thus, the use of recombinant erythropoietin seems to be safe. Our results suggest that negative feedback regulation persists between haemoglobin and erythropoietin in colorectal cancer. Production of erythropoietin remains therefore anaemia-associated, hypoxia-dependent and doesn't seem to be autonomic despite abundant expression of erythropoietin by colorectal cancers.

Increased expression of connexins 26 and 43 in lymph node metastases of breast cancer.

BACKGROUND: Gap junctions are intercellular channels composed of connexins, which mediate the direct passage of small molecules between neighbouring cells. They are involved in regulation of cell cycle, cell signalling, and differentiation, and probably invasion and metastasis. The role of connexins in the metastatic process is controversial, because some studies indicate that connexin expression is inversely correlated with metastatic capacity. In contrast, others demonstrate that connexins may be involved in metastasis. In addition, connexin status in breast cancer metastasis has not been widely studied. METHODS: We evaluated by immunohistochemistry the expression of connexin 26 (Cx26) and connexin 43 (Cx43) in primary breast tumours (PTs) and matched paired metastases to lymph nodes (MLNs). RESULTS: In PTs, we observed predominantly cytoplasmic localisation of evaluated connexins, indicating alterations in connexin expression in breast cancer cells. We demonstrated that expression of Cx26 and Cx43 was increased in MLNs compared with PTs (p<0.00001 and p<0.001, for CX26 and Cx43, respectively). In addition, Cx26 and Cx43 negative PTs developed Cx26 and Cx43 positive MLNs. Furthermore, besides increased cytoplasmic staining, enhanced membranous localisation of Cx43, typical of normal cells, was found in MLNs. Additionally, membranous Cx26 expression appeared only in metastatic breast cancer cells. CONCLUSIONS: These findings suggest that connexins may contribute to the efficient metastasising of breast cancer to the lymph nodes.

P53 correlates positively with VEGF in preoperative sera of colorectal cancer patients.

Diversity of P53 impact on tumor angiogenesis is due to the fact that wild-type P53 decreases expression of vascular endothelial growth factor (VEGF), but mutant P53 upregulates it. Therefore, we aimed at uncovering relations between preoperative serum levels of VEGF and P53 in colorectal cancer (CRC) patients. Preoperative blood samples of 125 CRC patients and 16 control healthy volunteers were examined with an ELISA-kit for serum P53 levels and VEGF. P53 did not correlate with VEGF in the whole group of CRC patients. However, P53 associated with VEGF in case of colorectal cancer patients, whose serum values of VEGF were higher than in controls (VEGF{H} >5.9333 pg/ml) (r=0.274, p<0.009). We revealed a positive correlation between P53 and VEGF{H} in subsets of poorly differentiated (G3) cancers (p<0.02), lymph node positive (p<0.007), pT3 or pT4 patients (p<0.004) without analogous relation in moderately differentiated (G2) tumors, node negative patients or pT1 or pT2 patients. P53 and IGF-I negatively correlated in all CRC patients (p<0.04) and VEGF{H} individuals of pT3 or pT4 (p<0.05) without any significant linkage in tumors of pT1 or pT2. The positive correlation between serum P53 and VEGF points at mutation of P53 and is a highly probable sign of poor prognosis in colorectal cancer. For now it can not be excluded that the binary analysis of serum P53 and VEGF could help select CRC patients endangered by rapid growth and lymph node metastases.

To give or not to give recombinant EPO to anemia endangered cancer patients.

EPO is known as an inducer of maturation and proliferation of erythrocytes. Moreover, it favours angiogenesis. In several studies it was encountered that EPO is a trophic agent that mediates survival and inhibits apoptosis of hypoxia affected cells, particularly those which build masses of irregularly vascularized cancers. The main task concerning EPO for oncologists is the choice to give or not to give recombinant EPO to anemia endangered cancer patients. EPO can do the quality of life better and cause recovery from anemia post chemotherapy and radiation of cancer patients. Nevertheless, EPO therapy shortens survival of patients in some cancers, in which antiapoptotic effect of EPO predominates directly in malignant cells. Thus, separately in every type of cancer, therapeutic use of recombinant EPO calls for prior investigations, if EPO signaling causes proliferation of cancer cells by direct stimulation of EPOR positive malignant cells. Unless the proliferative effect of EPO on cancer cells is excluded, its use in the therapy of anemia in cancer patients is not quite safe.

Expression of insulin receptor substrate 1 in primary breast cancer and lymph node metastases.

BACKGROUND: Insulin receptor substrate 1 (IRS-1) transmits signals from the insulin-like growth factor I receptor (IGF-IR) and insulin receptor (IR) and has been associated with the pathogenesis of cancer. IRS-1 downregulation has been suggested to play a role in breast cancer progression, but no simultaneous assessments of IRS-1 expression in primary breast cancer and metastases have been performed. AIMS: To assess IRS-1 expression in primary and metastatic breast cancer. METHODS: IRS-1 expression was analysed by means of immunohistochemistry in 109 samples of primary breast cancer and in 42 matched primary and metastatic tumours. In addition, IRS-1 expression was correlated with selected clinicopathological features, including oestrogen receptor alpha (ERalpha) and proliferation marker Ki-67 status. RESULTS: Positive cytoplasmic IRS-1 immunostaining was found in 69.7% (76 of 109) and 76.2% (32 of 42) of the primary and metastatic tumours, respectively. Both IRS-1 positive and IRS-1 negative primary tumours produced IRS-1 positive and IRS-1 negative metastases. IRS-1 expression in primary tumours correlated with poorly differentiated (G3) breast cancer (p < 0.005) and with lymph node involvement (p <0.05). In the subgroup of ERalpha positive primary tumours, IRS-1 expression positively correlated with Ki-67 (p < 0.02, r = 0.351), but in the subgroup of ERalpha negative primary tumours there was a negative correlation (p < 0.03, r = -0.509). IRS-1 expression in lymph node metastases correlated with neither ERalpha nor Ki-67. CONCLUSIONS: IRS-1 might be involved in breast cancer progression. Knowledge about differences between primary and metastatic tumours might help to understand mechanisms of breast cancer progression and lead to the development of more effective anticancer drugs.

Expression of the Insulin Receptor Substrate 1 in primary tumors and lymph node metastases in breast cancer: correlations with Bcl-xL and Bax proteins.

In our previous investigation Insulin Receptor Substrate 1 (IRS-1) correlated with proliferation marker Ki-67 in human breast cancer. The aim of the present study was to assess relationships between IRS-1 expression and anti-apoptotic Bcl-xL as well as proapoptotic Bax proteins, assessed by immunohistochemistry, in primary tumors and lymph node metastases of breast cancer. IRS-1 is positively associated with both Bcl-xL and Bax in primary and metastatic tumors. Thus, our results could suggest that IRS-1 might affect turnover of cancer cells and breast cancer progression through activation of mitogenesis and participation in the regulation of the balance between anti- and proapoptotic pathways.

The activity of cancer procoagulant in cases of uterine leiomyomas.

PURPOSE: It is currently believed that cancer procoagulant (CP), an enzymatic protein, is a product of malignant neoplastic cells. The present study was designed to test whether it is also synthesized by benign neoplastic cells, namely uterine leiomyomas. MATERIALS AND METHODS: We determined the activity of CP in the blood serum of women with uterine leiomyomas (N = 24), normal women (N = 15), and genital cancer patients (N = 6) by the coagulative method according to Gordon and Benson. Also, the CP activity in 10% tissue homogenates of uterine leiomyomas, normal uterine muscle and tissues of cervical and endometrial carcinoma was determined by the chromogenic method according to Colucci et al. RESULTS: The mean CP activity in the sera of women with uterine leiomyomas was 181.1 seconds (s) +/- 19.9 s, in healthy women--293.2 s +/- 33.8 s, and in genital cancer patients--78.8 +/- 18.5 s (all differences: p < 0.001). Similarly, in homogenates of uterine leiomyomas the CP activity was 19.6 +/- 3.8 nmoles pNa/ml, in normal uterine muscle it was 13.2 +/- 2.2 nmoles pNa/ml, and in cancerous tissue--28.0 +/- 6.6 nmol pNa/ml (all values being significantly different from each other). There was a strong correlation (r = -0.8122; p < 0.001) between the CP activity in uterine leiomyomas and serum activity, suggesting that the source of the serum CP activity was from the leiomyoma. The coagulation time of 120 to 240 s by the Gordon and Benson method supported the diagnosis of uterine leiomyoma, and a value below 120 s--the suspicion of genital cancer. CONCLUSIONS: Uterine leiomyomas, representing benign genital neoplasia, synthesize CP and are the likely origin of CP activity in blood, as has been described for malignant tumors, but to a lesser degree. There may be a role for CP as a tumor marker of genital neoplasia.

The contribution of type II pneumocytes and alveolar macrophages to fibroplasia processes in the course of enzymatic lung injury.

The aim of the paper was to evaluate mutual relations in the system of alveolar macrophage (AM) - type II pneumocyte (PII) - interstitium of alveolar septa, in the course of experimental lung emphysema in rats subjected to BCG vaccine effect. Administration of BCG vaccine resulted in the cumulation of AM within pulmonary alveoli. These cells exhibited morphological features of increased activity. Intratracheal papain injection induced intralobular emphysema changes, partly generalized, in the animal lungs. The emphysematous changes, with domination of interalveolar septum atrophy, were accompanied by focal accumulation of collagen and elastin. Fibroplasia processes were strongly pronounced in BCG- and papain-treated animals. The areas of connective tissue fibres cumulation revealed indistinctness of the boundary line between PII and the interstitium in some places. Anchorage of collagen fibres and microfibrillary structures were observed in the cytoplasm of PII. The morphological examinations of AM - fibroblasts co-cultures as well as the evaluation of the uptake of 3H-thymidine did not show any significant differences between respective co-cultures of fibroblasts and AM isolated both from the lungs of control and experimental animals (treated with BCG or papain, and BCG+papain). However, a significant growth was noted in 3H-thymidine uptake between fibroblast cultures realized with or without cells isolated from the lungs. The results obtained suggest the possibility of active participation of PII and AM in fibroplasia processes in the course of lung rebuilding after papain administration and in pathological states of the pulmonary tissue, particularly when they are accompanied by increased activity of alveolar macrophages. They also support the inflammatory-repair hypothesis in the development of emphysematous changes.

Alveolar cells in cyclophosphamide-induced lung injury. II. Pathogenesis of experimental endogenous lipid pneumonia.

An ultrastructural and histological study was made to analyse the structural and cellular features of the pulmonary lesions produced in Wistar rats by intraperitoneal (i.p.) administration of cyclophosphamide (two i.p. doses of 150 mg CP/1 kg bw/1 ml PBS). Rats exposed to cyclophosphamide (CP) developed a condition whose morphological picture corresponded to endogenous lipid pneumonia and/or pulmonary alveolar proteinosis-like changes. Damage to the endothelium and neutrophil accumulation in lung vascular bed were found to be potential initiators of endogenous lipid pneumonia-type changes. The possibility of the evolution of the acute lung injury into endogenous lipid pneumonia-type changes and into alveolar proteinosis-like changes was demonstrated. The results of the study supplement the existing theories of pulmonary alveolar proteinosis pathogenesis.

Alveolar cells in cyclophosphamide-induced lung injury. An ultrastructural analysis of type II alveolar epithelial cells in situ.

Recent studies have brought rich evidence in favour of the significant contribution of the surfactant system-forming structures to morphogenesis of many pulmonary disorders. The aim of this study was to evaluate the effect of intraperitoneal cyclophosphamide administration on changes within this system. The experiments used 40 Wistar rats, of 170 g body weight. The animals were divided into two experimental groups. Group I animals were given cyclophosphamide (Endoxan-ASTA) in a single intraperitoneal dose of 150 mg/1 kg b.w./1 ml PBS/. Group II (control) received 1 ml PBS. All the animals were sacrificed after 1, 3, 7 and 28 days following intraperitoneal cyclophosphamide or PBS administration. Morphological examinations of pulmonary tissue were based on ultrastructural analysis in the transmission electron microscope. The study revealed that a single intraperitoneal cyclophosphamide injection caused damage to all elements forming the surfactant system, particularly to type II alveolar epithelial cells. Rebuilding processes in pulmonary tissue, coexisting with destructive changes, occurred with a significant contribution of type II alveolar epithelial cells. These cells are likely to take an active part in pulmonary fibrosis processes observed after the action of cyclophosphamide.

The effects of cyclophosphamide on pulmonary thrombopoiesis in rats.

Cyclophosphamide (CP), an antineoplastic and immunosuppressive agent, even when administered in large doses, slightly affects the quantity of blood platelets. The aim of the present study was to analyse the effect of single intraperitoneal administration of CP (150 mg/kg b.w.) on the quantitative changes in platelets obtained from the left and right ventricle of the heart, as well as to evaluate the occurrence of megakaryocytes in lung tissue depending on the period of time that passed from CP administration. In control subgroups, fewer platelets were found in the blood collected from the RV compared with the left ventricle at all time intervals. After 1 and 3 days following i.p. administration of CP, a decrease was observed in the number of platelets both in the blood from the right ventricle and left ventricle when compared with control. However, after 14 days, the number of platelets in the blood from the left ventricle was higher, compared with the left ventricle and right ventricle of control animals, and significantly higher (p < 0.001747), compared with their number obtained from the right ventricle of CP-receiving animals. Simultaneous ultrastructural examinations with transmission electron microscopy revealed the increased number of platelets in the lung vascular bed of CP-receiving rats at all time intervals. However, megakaryocytes were found 7 and 14 days after administration of CP. The findings clearly indicate that the lungs could be a major place of thrombopoiesis following therapy with a single large dose of CP.

Type II alveolar epithelial cells and free alveolar cells after intratumor TNF-alpha administration.

The experiment used Morris hepatoma 5123 series growing in muscles of the Buffalo rats. A suspension of 3 x 10(6) neoplastic cells was injected into the right hind leg of the animals. After fourteen days, TNF-alpha was administered into the tumour in a dose of 1.5 x 10(4) U/24 hours in 0.5 ml PBS solution. The group I animals were injected for 4 days and group II for 8 days. Control groups consisted of rats with injected Morris hepatoma which were given PBS solution instead of TNF-alpha (group III A and B) and animals without the hepatoma, given 4 or 8 TNF-alpha, respectively (groups IV A and B). In the present study, we have explored the effect of intratumor TNF-alpha administration on the composition of cells isolated from the lungs through multiple bronchoalveolar lavages (BAL). Ultrastructural evaluation of the pulmonary tissue was done using a transmission electron microscope (TEM), with special attention paid to type II alveolar epithelial cells and free alveolar cells. Examinations in TEM in groups I, II and IV (A and B) found, in the lumen of alveoli, an increase in the number of alveolar macrophages (AM) with morphological features of intensified activity and AM with numerous secondary lysosomes containing material of phospholipid structure. Also, numerous type II alveolar epithelial cells with emptied lamellar bodies were observed. The above mentioned changes were especially marked after eightfold TNF-alpha administration. In groups I, II and IV (A and B), compared with group III, a significant increase was found in the total number of cells isolated by BAL as well as in the number of cells with positive reaction in staining according to Beckstead's method. It may indicate that the changes in the parameters mentioned above are related to TNF-alpha action. The results obtained indicate the possibility of systemic effect of TNF-alpha after its administration into the experimental Morris hepatoma.

Regression of Morris hepatoma in response to intralesional treatment with tumor necrosis factor muteins.

We examined the antitumor effects of human recombinant tumor necrosis factor alpha (rhTNF-alpha) and its muteins with the N-terminal amino acid sequence altered by point mutations against transplantable Morris hepatoma 5123 in rats. In vivo studies showed antiproliferative activity of the drugs in the dose range tested. For in vivo studies rhTNF-alpha and muteins were administered intratumorly (i.t.). The preparations were given at a dose of 10 microg/rat, once daily for eight days. Although the therapy was significantly effective in inhibiting tumor growth, complete growth inhibition could not be achieved. Nevertheless, there was a significant increase in survival time of tumor-bearing rats.

Alveolar cells in cyclophosphamide-induced lung injury. The effect of pentoxifylline on BAL-isolated cells.

The present study provides the evaluation of pentoxifylline (PTXF) effect on quantitative and morphological changes of cells isolated from the lungs by BAL method in animals given cyclophosphamide (CP). CP was administered once, intraperitoneally, on the 7th day of the experiment, in a dose of 150 mg/kg b.w. PTXF was administered in drinking water in a dose of 30 mg/kg b.w./24 h for 14 days of the experiment. Morphological examinations of the isolated cells were performed using the scanning electron microscope (SEM). An attempt to identify type II alveolar epithelial cells was also undertaken. A single CP dose caused a significant rise in the total number of cells isolated from the lungs of the animals 7 days following CP administration. At the same time, a fall was noted in the percentage of neutrophilic granulocytes. No significant quantitative changes were observed in the population of cells isolated after 1, 3 and 28 days following CP administration. PTXF, in doses applied, did not significantly affect quantitative changes and morphological picture of the isolated cells. Our observations suggest that none of the methods applied for identification of type II pneumocytes are completely specific. At the same time they stain type II cells and subpopulations of alveolar macrophages heavily loaded with phagocytized surfactant. However, those methods are simple and enable an approximate evaluation of damage and regeneration processes in the extracellular alveolar lining layer and epithelium.

Ultrastructural study of endogenous lipid pneumonia and pulmonary alveolar proteinosis coexistent with lung cancer.

The aim of the study was the ultrastructural analysis of endogenous lipid pneumonia (ELP)-type changes localized in the vicinity of primary neoplasms of the lungs. The analysis was carried out on the material obtained from 30 patients with non-small cell lung carcinoma (NSCLC). Routine histopathological examination was performed in light microscope, focusing mainly on the presence or lack of ELP-type changes. Ultrastructural examinations in transmission electron microscope used 12 cases classified among early (4 cases) or advanced (8 cases) ELP forms. Results of the study reveal a likely coexistence of ELP- and PAP (pulmonary alveolar proteinosis)-type changes in the vicinity of NSCLC and suggest a correlation between the processes leading to the development of ELP or/and PAP. Ethiopathogenesis of the changes observed and the role of granulocytes, macrophages and type II alveolar epithelial cells are discussed.

Stimulation of alveolar macrophages by BCG vaccine enhances the process of lung fibrosis induced by bleomycin.

It was found that the BCG vaccine injected subcutaneously to the rats enhances the process of lung fibrosis induced by bleomycin. Pretreatment of rats with this vaccine results in accumulation of activated macrophages in lung interstitium and in the bronchoalveolar spaces. It may be suggested that the activated macrophages release various cytokines which may stimulate the proliferation of fibroblasts and biosynthesis of extracellular matrix components.