Analysis of hip joint loading during walking with different shoe types using instrumented total hip prostheses.

Hip joint loads need careful consideration during postoperative physiotherapy after joint replacement. One factor influencing joint loads is the choice of footwear, but it remains unclear which footwear is favorable. The objective of the present study was to investigate the influence of footwear on hip joint loads in vivo. Instrumented hip endoprostheses were used for in vivo load measurements. The parameters resultant contact force (Fres), bending moment (Mbend) and torsional moment (Mtors) were evaluated during treadmill walking at 4 km/h with different shoe types. In general, footwear tended to increase hip joint loading, with the barefoot shoe having the least influence. Fres and Mbend were significantly increased during heel strike for all shoe types in comparison to barefoot walking, with everyday shoe (34.6%; p = 0.028 and 47%; p = 0.028, respectively) and men's shoe (33.2%; p = 0.043 and 41.1%; p = 0.043, respectively) resulting in the highest changes. Mtors at AbsMax was increased by all shoes except for the barefoot shoe, with the highest changes for men's shoe (+ 17.6%, p = 0.043) and the shoe with stiffened sole (+ 17.5%, p = 0.08). Shoes, especially those with stiff soles or elaborate cuishing and guiding elements, increase hip joint loads during walking. The influence on peak loads is higher for Mtors than for Fres and Mbend. For patients in which a reduction of hip joints loads is desired, e.g. during physiotherapy after recent surgery or to alleviate symptoms of osteoarthritis, low profile shoes with a flexible sole may be preferred over shoes with a stiff sole or elaborate cushioning elements.

Anastomosing hemangioma of the kidney: radiologic and pathologic distinctions of a kidney cancer mimic.

Anastomosing hemangioma (ah) is a rare subtype of primary vascular tumour that can, clinically and radiologically, present similarly to malignant renal tumours such as renal cell carcinoma (rcc) and angiosarcoma. Rarely seen in the genitourinary system, the ah we report here occurred in a 40-year-old male patient diagnosed initially with rcc based on imaging and successfully treated by laparoscopic left radical nephrectomy, with adrenal sparing and perihilar lymph node dissection. The pathologic diagnosis of ah can be challenging on small biopsy specimens; we therefore opine that it is appropriate to excise these lesions to facilitate diagnosis and definitively exclude common renal cancers. However, in this review, we describe some radiologic and pathologic distinctions between ah and malignant tumours.

Underexpression of tumour suppressor LKB1 in clear cell renal cell carcinoma is common and confers growth advantage in vitro and in vivo.

BACKGROUND: Evidence suggests that dysregulation of energy-sensing pathways closely associates with renal cell carcinoma (RCC) development. The metabolic regulation is largely controlled by 5'-AMP activated protein kinase (AMPK) which is activated through phosphorylation by LKB1. METHODS: The expression of LKB1 was determined by reverse transcription-PCR using 10 clinical clear cell RCC (ccRCC) samples and their adjacent normal renal parenchyma, and by immunohistochemical staining of two tissue microarrays containing 201 ccRCC and 26 normal kidney samples. Expression of LKB1 was knocked down in human ccRCC 786-O cells (shLKB1) and compared with cells expressing scrambled control shRNA (shControl). AMPK signalling, proliferation, invasion, and VEGF secretion was measured. The cells were subcutaneously injected into mice to determine tumour growth in vivo. RESULTS: At the protein and transcript levels, a significant reduction in LKB1 expression in tumour compared with normal tissue was found. In vitro, knockdown of LKB1 resulted in reduced AMPK signalling and increased cellular proliferation, invasion, and VEGF secretion compared with shControl cells. In vivo, growth of shLKB1 ccRCC xenografts in nude mice was significantly increased compared with shControl xenografts. CONCLUSION: Collectively, our results suggest that LKB1 acts as a tumour suppressor in most sporadic cases of ccRCC and that underexpression of LKB1 is a common event in the disease.

PSA affects prostate cancer cell invasion in vitro and induces an osteoblastic phenotype in bone in vivo.

BACKGROUND: Patients with advanced prostate cancer frequently have a poor prognosis as a result of metastasis and present with high serum PSA levels. There is evidence suggesting that the serine protease activity of PSA could be involved in the invasion and metastasis of prostate cancer. In this study, we determined the effects of PSA and its precursor, pro-PSA, on invasion and the type of bone metastasis. METHODS: We stably transfected prostate adenocarcinoma cells, human DU-145 and rat MatLyLu, with either the full-length prepro-PSA sequence or pre-PSA DNA, to generate subclones of cells that secrete pro-PSA or free PSA, respectively. Secretion of PSA was measured by western blot analysis and enzyme-linked immunosorbent assay (ELISA). The invasive and migratory properties of the cells were determined using a basement membrane extract and were compared with corresponding empty vector control cells. Twelve days after injection of PSA-secreting MatLyLu cells into the femora of nude mice, bone tumor burden and histomorphometry were determined using a stereological technique. RESULTS: The transfected cells secreted 0.15-2.23 ng PSA/10(6) cells/day. Pro-PSA-secreting subclones increased invasion and migration by 24-263%. Conversely, the PSA-secreting subclones significantly reduced both invasion and migration by 59-70%. The divergent effects on invasion and migration observed in pro-PSA- and PSA-secreting subclones indicate that different forms of PSA may have different functions. Intrafemoral injections with PSA-secreting MatLyLu cells resulted in an increase in osteoblastic parameters when compared with non-PSA-secreting subclones as measured by bone histomorphometry. Concomitantly, a decrease in osteoclasts and eroded surface was observed. CONCLUSIONS: Our in vitro data suggest that PSA, dependent on the predominant form secreted, may decrease or increase invasive properties of prostate cancer cells. The in vivo results indicate that PSA in the bone microenvironment may contribute to the osteoblastic phenotype of bone metastasis frequently observed in prostate cancer.

Expression analysis of genes involved in epigenetic regulation and apoptosis in human malignant haematopoietic cell lines treated with 5-azacytidine.

PURPOSE: The aim of this study was to investigate the modulation of the expression status of 10 different genes involved in epigenetic regulation and apoptosis by the DNA methyltransferase (DNMT) inhibitor 5-azacytidine (5-Aza), as markers of response to treatment, in two different human malignant haematopoietic cell lines. METHODS: In our analysis we used the SybrGreen technology and gene-specific primers for the qRT-PCR analysis of 10 genes, in cDNA of PC-MDS and K562 cell lines, treated by 1 micromole of 5-Aza for 24h. RESULTS: DNMT1 and DNMT3A showed statistically significant decrease of expression in 5-Aza-treated PC-MDS cells, whereas DNMT3B showed significantly decreased expression in 5-Aza-treated K562 cells. The members of the Bcl- 2 family of apoptosis-regulating genes Bcl-2 and Bax showed statistically significant differences in expression, in comparison with non-treated PC-MDS cells. Our most interesting result was the significant upregulation (re-expression) of p15, in 5-Aza-treated PC-MDS cells. CONCLUSION: The re-expression of p15 in PC-MDS cell line evaluated by qRT-PCR makes this novel cell line a suitable model for the studies of pharmacologic demethylation as a plausible mechanism resulting in hematologic response in myelodysplastic syndrome (MDS).

Cardiac autonomic control in patients with myasthenia gravis and thymoma.

OBJECTIVE: To evaluate cardiac autonomic control in patients with myasthenia gravis (MG) and thymoma. MATERIALS AND METHODS: The study was performed on 21 patients with MG and thymoma and the same number of matched healthy volunteers. Standard cardiovascular reflex tests according to Ewing and baroreflex sensitivity (BRS) at rest was applied. Spectral analysis of heart rate variability (HRV) at rest was assessed using a 20-minute ECG recording (normalized low- and high-frequency bands-LFnu-RRI, HFnu-RRI and LF/HF-RRI) Time-domain analysis of HRV was derived from 24-hour ECG monitoring. RESULTS: Overall autonomic score according to Ewing was significantly increased in patients with MG and thymoma (p<0.05), mostly due to parasympathetic dysfunction. Time-domain parameters representing the overall and long-term sympathetic activity of HRV did not differ significantly between the two groups (p>0.05), but there was a significant decrease in measures of the short-term vagal variations in HRV (p<0.01). HFnu-RRI was lower, while LFnu-RRI and LF/HF-RRI were higher in patients with MG and thymoma in comparison to healthy controls but these differences were not of statistical significance (p>0.05). BRS at rest was highly significantly reduced in patients group (p<0.01). CONCLUSIONS: Our results showed mainly parasympathetic cardiac impairment in patients with myasthenia gravis and thymoma. Since autonomic dysfunction may lead to cardiac conduction abnormalities and sudden death, the investigation of autonomic nervous system function in these patients may be significant in everyday clinical practice.

Quantitative real time-polymerase chain reaction method in Bcr-Abl translocation diagnostics.

PURPOSE: The quantitative real-time polymerase chain reaction (qRT-PCR) is used in the detection of molecular events involved in leukemogenesis, such as the Bcr-Abl gene translocation, the most important factor in the pathogenesis of chronic myeloid leukaemia (CML). The main aim of our study was to test the reproducibility, specificity and sensitivity of the qRT-PCR in the detection of Bcr-Abl gene translocation. METHODS: In complementary (c)DNA, isolated from K562 Bcr-Abl positive cell line, we performed qRT-PCR analysis with Bcr-Abl specific primers. For qRT-PCR analysis, we used serial dilutions of the newly synthesized cDNA in order to establish the detection threshold of this method. RESULTS: Using the specific primers for the Bcr-Abl translocation, we obtained the specific translocation product in cDNA sample of K562 human erythroid leukemia cell line. qRT- PCR showed significant sensitivity with the detection threshold for the Bcr-Abl fluorescent signal, which enabled the precise detection that was accurate within a 10-fold dilution range, and a dynamic range of 5 orders of magnitude. CONCLUSION: The results of our study showed that the application of the qRT-PCR is the optimal method for the detection of Bcr-Abl gene translocation, characterized by high reproducibility, specificity and sensitivity.

Antioxidant enzymes activities and plasma levels of oxidative stress markers in B-chronic lymphocytic leukemia patients.

PURPOSE: Overproduction of reactive oxygen species (ROS) intermediates above the functional capability of cellular antioxidants may result in instability of important macromolecules and represents the molecular basis of many diseases including inflammation processes, cardiovascular alterations, cancer etc. The purpose of this study was to determine plasma level of superoxide anion, hydrogen-peroxide and malondialdehyde (MDA) as markers of oxidative stress and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) as antioxidant enzymes in B-chronic lymphocytic leukemia (B-CLL) patients. METHODS: The study included 29 untreated B-CLL patients in stage A, and 21 in stages B and C, classified according to the Binet system; 31 healthy volunteers formed the control group. After centrifugation of heparinized peripheral blood, plasma levels of all investigated parameters were determined using spectrophotometric methods. RESULTS: Plasma CAT activity was increased in B-CLL patients compared with control subjects; also, progression of disease was related with significantly higher plasma activity of CAT. Also, B-CLL patients showed significantly higher plasma concentration of MDA compared with controls. No statistically significant differences of superoxide anion and hydrogen peroxide as well as plasma activity of SOD and GPx between the tested groups were noted. CONCLUSION: Increase of CAT activity in B-CLL patients indicates that there is stimulation of the antioxidant enzyme system, while the increase of MDA concentration shows increased lipid peroxidation level. According to these results it could be concluded that an imbalance exists between oxidants and antioxidants in the plasma of B-CLL patients.

In vitro induction of apoptotic cell death in chronic lymphocytic leukemia by two natural products: preliminary study.

PURPOSE: B cell chronic lymphocytic leukemia (B-CLL) is an neoplastic disorder characterized by alterations in the pathways of programmed cell death (apoptosis). Deregulation of apoptosis pathways also contributes to chemoresistance of B-CLL cells. Therefore, it is not surprising that induction and acceleration of apoptosis represent key point in novel B-CLL therapeutic protocols. The present study was designed to investigate the effects of two natural products, Immunarc forte and Korbazol on the in vitro survival of leukemic cells. METHODS: peripheral blood mononuclear cells (PBMC) from 20 B-CLL patients and 20 healthy donors were used for cytotoxicity studies. Cytotoxic activity of the tested products were assessed by the MTT colorimetric assay and the type of cell death was determined by flow cytometry. RESULTS: we found that Korbazol was selectively cytotoxic against B-CLL cells, but the cytotoxic activity of Immunarc forte was much weaker. Of note, synergy was shown between these two drugs, and this effect was also selective, without affecting the normal mononuclear cells. According to Annexin-V binding, Korbazol and Immunarc forte induced apoptotic type of cell death in B-CLL cells. Moreover, treatment with Korbazol, but not with Immunarc forte, decreased spontaneous apoptosis in cultured normal polymorphonuclear cells. CONCLUSION: our findings imply that Korbazol is as potential therapeutic agent that induces apoptosis of B-CLL cells. The resistance of normal mononuclear cells and anti-apoptotic effects on normal polymorphonuclear cells, as well as its ability to synergize with Immunarc forte, warrants further investigation and supports their therapeutic application in the treatment of B-CLL.

Endoplasmic reticulum stress associated with caspases-4 and -2 mediates korbazol-induced B-chronic lymphocytic leukemia cell apoptosis.

PURPOSE: B-cell chronic lymphocytic leukemia (B-CLL) is an incurable disease that rapidly develops drug resistance. Therefore there is a need for identifying new agents that will improve the therapeutic outcome. Korbazol is a natural product known to exert cytotoxic effect on the in vitro survival of leukemic cells. The aim of this study was to investigate the mechanism of korbazol-induced apoptosis in B-CLL leukemic cells. METHODS: peripheral blood mononuclear cells from 10 B-CLL patients were used for assessing the effect of caspase inhibitors and chelator of intracellular Ca(2)+. RESULTS: cell death rate induced by the tested compound was decreased with the caspase-3 inhibitor Ac-DEVD-CHO, and the inhibitors of caspase-2 (Z-VDVAD-FMK) and -4 (ZYVAD- FMK), but not with the caspase-9 inhibitor z-LEHD-FMK and caspase-8 inhibitor z-IETD-FMK. No significant release of cytochrome C (cyt C) from mitochondria to the cytosol of B-CLL cells treated with korbazol was observed. Moreover, chelating of intracellular Ca(2)+ with BAPTA-AM almost completely abolished the cytotoxic effect of korbazol. CONCLUSION: engagement of caspases-2 and -4 and mobilization of intracellular Ca(2)+ indicate involvement of endoplasmic reticulum (ER) stress in apoptosis induced by korbazol.

Oxidative stress accelerates spontaneous apoptosis of B-chronic lymphocytic leukemia lymphocytes.

PURPOSE: B-chronic lymphocytic leukemia (B-CLL) is characterized by the progressive accumulation of small immature B lymphocytes which do not undergo apoptosis due to an underlying defect. One potential mechanism of defective apoptosis could be irregular oxidative stress. The goal of our investigation was to determine in vitro production of oxidative stress markers by lymphocytes of B-CLL patients. PATIENTS AND METHODS: 30 untreated stage A B-CLL patients, as well as 20 stage B and C patients and 30 healthy volunteers as a control group were examined. Nitric oxide (NO), superoxide anion, hydrogen peroxide and malondialdehyde (MDA) were measured by spectrophotometry in supernatants of lymphocytes cultures of all 3 investigational groups. The method applied for detecting apoptosis was fluorescence microscopic analysis using acridine orange/ethidium bromide (AO/EB) double staining. RESULTS: In vitro lymphocyte production of superoxide anion, hydrogen peroxide and MDA was increased in B-CLL patients, while there were no statistical significantly differences of NO production among the tested groups. Compared with the spontaneous apoptosis observed in control subjects lymphocytes, B-CLL lymphocytes showed increased percentages of apoptotic cells after incubation for 24 h. Disease progression was not followed with significant differences in spontaneous apoptosis of B-CLL lymphocytes. CONCLUSION: This intensive oxidative stress markers production in cultures of B-CLL lymphocytes could be one of the potential mechanisms in the pathogenesis of abnormal apoptosis.

Discordance between receptor status in primary and metastatic breast cancer: an exploratory study of bone and bone marrow biopsies.

AIMS: The treatment of bone metastases in breast cancer is traditionally based upon the receptor status of the primary tumour. However, retrospective studies have shown significant discordance in receptor expression between primary and metastatic tumours. Therefore, the aim of this study was to prospectively assess the incidence of discordant receptor status in primary and metastatic disease and evaluate the role of bone marrow biopsies for the reassessment of receptor status. MATERIALS AND METHODS: Nine patients with known bone metastases were assessed with both a radiologically guided bone biopsy and a bone marrow aspirate and trephine. The oestrogen receptor and progesterone receptor status of these samples was assessed and compared with the primary breast cancer. Bone and bone marrow samples were also evaluated for HER2/neu status and compared with the status of the primary tumour if available. RESULTS: Tumour cells were found in six of the nine bone metastasis specimens and five of the nine bone marrow samples. A discordance rate for the oestrogen receptor was seen in five of nine patients (56%) and for the progesterone receptor in four patients (44%). There seemed to be a correlation between bone and bone marrow biopsies. CONCLUSION: The receptor discordance rate in this study was similar to previous retrospective studies. It seems that bone marrow biopsy may be a simple, safe and well-tolerated way to obtain tissue to reassess the receptor status of metastatic breast cancer.

[Carotid artery plaque in patients with disorders of glucose regulation].

PURPOSE: The aim of this study was measurement of artery intima media thickness (IMT) and plaques as an early indicator of atherosclerosis in diabetics comparing with other risk factors of carotid artery. METHODS AND MATERIALS: 110 pts: 50 with Diabetes Mellitus, type 1 (25) and type 2 (25), 20 pts with glucose intolerance, 20 pts with type 2 de novo and 20 pts obese without diabetes. Ultrasound examination (using 7.5 MHz sound on Toshiba SSA-270A) end measurement of intima-media ticknes (IMT) were performed on Carotis communis (CCA), bifurcation and distal from bifurcation to a.carotis intern (ACI), expressed in mm. Plaques were correlated with other common factors age, BP, lipid parameters (Chol, HDL, LDL, Triglycerides), smoks, alcoholism and obese (BMI). The authors used 2 test and Spearman's correlation. RESULTS: The lowest percent of plaques was found in group with type DM 1. The highest percent of plaques was found in type DM 2. Statistically there is highly significant difference between plaques founded on type 2 DM and types 2 DM de novo and on other types. CONCLUSION: DM is not an independent risk factor for developing of macroangiopathic changes an arterial walls, but their appirience are more presenting in diabetic patients. The highest number of plaques are presenting DM type 2 (29.6%), and after type 2 de novo (26.8%), the next highest position of plaques were in patients with obese but without DM and intolerantio glucosae (IFG+IGT) (17.1%) and type 1 DM (9.8%). Risk factors were presented in following percentage: Obese 80.5% pts; hyperlipidema 53.7% pts; HTA 51.3%; smoking 51.2% pts and alchocholism 2.4% pts. According to these results, all risk factors were included in patophysiology of plack forming except alcoholism. Influences of these risk faktors are very importance and their synergic action lids to their rapid appirience and clinical manifestations. DM has specific position in patophisiology of atherosclerosis.

[Correlation of metabolic parameters and changes in the intima-media complex in non-diabetic obesity and patients with various disorders of glucose regulation].

UNLABELLED: Measurement of intima-media thickness (IMT) of carotid artery by ultrasound, is well known method. Using this noninvasive method in various risck factors such as diabetes mellitus, dislipidemia, hypertension, smoking, age, obesity could be very usefull in pathophisiology studies of atherosklerosis. PURPOSE: The aim of this study was to measure intima media thickness (IMT) of carotid artery, as an early indicator for development of atherosclerosis, and too estemate if there is any significant differences between investigated groups of pts (diabetics pts, pts with glucose intolerance and obese pts without diabetes an older than 45 years). METHODS AND MATERIALS: 110 pts were devided in five groups: three groups of diabetics pts: type 1 DM (25) pts, type 2 (25) pts and type 2 DM de novo (20), and (20) pts with glucose intolerance, and 20 pts obese without diabetes, older than 45 years of age. Correlation of metabolic parameters (body mass index (BMI), hip circumference (OS), lipid status) with IMT was performed as well as hipertension, age, sex, smoking and alcohol abuse. STATISTICS: The authors used 2 test and Spearman's correlation. RESULTS: Intima media ticknes (IMT) was highly statisticly significant in groups of DM type 2 and DM tipe 2 de novo. IMT was stat. sig. in pts with high values of BMI and hip circumference according to those pts with normal values. It was also stat. sig. in pts with smoking hiperlipoproteinemia and hipertension. There was no stat. sig. correlation of IMT with sex, age and alcohol abuse. CONCLUSION: This study showed that diabetics pts have frequent patological changes on arteri vessels and these changes are difuse and frequently with complicated wall leasens. The influence of these risck factors is very important because their sinergetic effects may augment clinical manifestation of atherosclerosis.

Effect of zoledronic acid on the doxycycline-induced decrease in tumour burden in a bone metastasis model of human breast cancer.

Bone is one of the most frequent sites for metastasis in breast cancer patients often resulting in significant clinical morbidity and mortality. Bisphosphonates are currently the standard of care for breast cancer patients with bone metastasis. We have shown previously that doxycycline, a member of the tetracycline family of antibiotics, reduces total tumour burden in an experimental bone metastasis mouse model of human breast cancer. In this study, we combined doxycycline treatment together with zoledronic acid, the most potent bisphosphonate. Drug administration started 3 days before the injection of the MDA-MB-231 cells. When mice were administered zoledronic acid alone, the total tumour burden decreased by 43% compared to placebo treatment. Administration of a combination of zoledronic acid and doxycycline resulted in a 74% decrease in total tumour burden compared to untreated mice. In doxycycline- and zoledronate-treated mice bone formation was significantly enhanced as determined by increased numbers of osteoblasts, osteoid surface and volume, whereas a decrease in bone resorption was also observed. Doxycycline greatly reduced tumour burden and could also compensate for the increased bone resorption. The addition of zoledronate to the regimen further decreased tumour burden, caused an extensive decrease in bone-associated soft tissue tumour burden (93%), and sustained the bone volume, which could result in a smaller fracture risk. Treatment with zoledronic acid in combination with doxycycline may be very beneficial for breast cancer patients at risk for osteolytic bone metastasis.

Relationship of serum levels of tumor markers with tissue expression of gene products in ovarian carcinoma.

PURPOSE: The aim of this prospective study was to determine serum levels and tissue expression of CA125, CA 15-3, p53, HER-2 and nm23 tumor markers, which are used in the detection and follow up of patients with ovarian carcinoma. PATIENTS AND METHODS: 19 patients with malignant and benign ovarian tumors were included in this study. Serum levels of CA125, CA 15-3 and p53 tumor markers were detected in preoperative and postoperative blood samples using ELISA technique. Tissue expression of p53, HER-2 and nm23 were examined using immunohistochemistry. RESULTS: All serum tumor markers were elevated in patients with ovarian carcinoma. Serum level of CA 15-3 was increased in patients with ovarian carcinoma (median 48.33 U/ml, normal range 0-36), while it was normal in patients with benign ovarian tumors (median 20.67 U/ml; p >0.05). CA125 serum values were strikingly increased in ovarian carcinoma (median 264.16 IU/ml, normal range 0-35) and benign ovarian tumors (median 119.59 IU/ml; p <0.05). Serum levels of p53 in patients with ovarian carcinoma were increased (median 0.69 U/ml, normal range 0-0.50) compared to patients with benign tumors (0.32 U/ml; p <0.05). Histological HER-2 overexpression was detected in 7 cases, including 4 with strong (score 3+ and 2+) and 3 with weak or no HER-2 expression (score 1+ and 0) in ovarian carcinoma tissue; in benign tumors HER-2 overexpression was detected in 1 case (p >0.05). Strong overexpression of p53 was detected in 3 cases with malignant and none with benign tumors (p >0.05); and strong overexpression of nm23 was detected in 5 cases with malignant and 2 with benign tumors (p >0.05). CONCLUSION: Serum levels of CA125, CA 15-3 and p53 are strikingly increased, as well as the expression of HER-2 and p53 in carcinomatous tissue. Detection and analysis of multiple tumor-specific markers in serum and tissue can give useful clinical information for the management of ovarian carcinoma and can also improve the sensitivity and specificity of these markers.

In vitro assay for the quantitative measurement of apoptotic lymphocytes phagocytosis by peripheral blood monocytes.

Currently used assays for the quantification of apoptotic cells uptake by phagocytes have several methodological problems. Our assay overcomes some of these problems. As a source of apoptotic cells we used peripheral blood lymphocytes obtained from the patients with chronic lymphoblast leukaemia. Apoptosis was induced by incubating cells with cycloheximide for up to 24 h. The assay was performed in suspension of peripheral blood mononuclear cells. For the visualisation of the phagocytes and phagocyted cells and discrimination of phagocyted from bound apoptotic cells we used Acridine orange/Ethidium bromide double staining. Here we offer a simple test which enables reliable measurement and it can show the difference of phagocytic potential between different individuals.

Preliminary study of mononuclear phagocytosis during breast cancer therapy.

PURPOSE: To evaluate the eventual changes in the number and phagocytic functions of blood monocytes in breast cancer patients during surgical treatment and chemotherapy. MATERIALS AND METHODS: The absolute and relative number of peripheral blood leukocytes and monocyte phagocytic functions were determined at the time of diagnosis (I), following surgery (II), during (III) and after chemotherapy (IV) in 30 patients diagnosed with breast cancer. The control group consisted of 30 age-matched healthy women. RESULTS: The mean number of monocytes was significantly lower in cancer patients at diagnosis, while they increased following surgery reaching the control values. There were no postchemotherapy changes in the number of monocytes. Monocyte phagocytic activity was decreased at the time of diagnosis. Following surgery, the capacity of phagocytosis (CP) recovered to normal values, but the index of phagocytosis (IP) remained decreased. During and after chemotherapy, as well as one year after surgery, the IP still remained decreased. CONCLUSION: Our results showed that some properties of monocytes' phagocytic activity in cancer patients were decreased at diagnosis, returning back to normal range after surgical therapy. However, time is needed to confirm whether the alteration of IP may provide additional information when monitoring breast cancer patients.

Morphological, histomorphometric, and microstructural alterations in human bone metastasis from breast carcinoma.

Bone is one of the most common sites of breast cancer metastasis. Metastases are often associated with bone destruction and are a major cause of morbidity. We examined structural bone changes induced by metastatic tumor in bone biopsies from 33 patients with metastatic breast carcinoma (20 from patients with pathological femoral fracture and 13 with no fracture) and 20 normal controls. In all metastatic biopsies bone remodeling was shown to be tumor volume-dependent. Bone resorption and bone formation were biphasic with both increasing at earlier stages of metastatic bone disease and decreasing later on. A comparison of patients with fracture and no fracture did not reveal statistically significant differences in the extent of bone destruction or trabecular thinning. Bone histomorphometry showed limited ability to explain the higher bone volume loss in fracture patients (decreases of 42% and 25%, respectively, in fracture and nonfracture patients compared with controls). However, changes in bone quality, including increased disconnectivity and decreased connectivity, as evaluated by node-strut analysis, suggested that there were more structural changes in the fracture compared with the nonfracture group. The nonfracture group included six patients with no radiological evidence of bone metastasis (occult metastasis). They showed a higher tumor volume and a twofold lower eroded surface compared with the rest of the group. The decrease in bone volume (14% lower than controls) was below the limit of X-ray detection. Because we observed no increase in osteoclast-related parameters and no correlation between osteoclast surface and eroded surface, we believe that, in occult metastasis, osteoclastic bone resorption is not an important factor in overall bone resorption. Quantitatively, the eroded surface in direct contact with tumor cells was threefold higher than the osteoclast surface in occult metastasis, whereas the rest of the metastatic group (27 of 33) showed predominantly osteoclast-mediated eroded surface. Node-strut analysis on occult metastasis revealed a significant increase in disconnectivity without a concomitant significant decrease in bone volume and trabecular thinning. We conclude that, in occult metastasis, bone resorption may be more osteoclast-independent and other mechanisms involving the tumor cells may be more prevalent.

[Association between HLA antigens and leukemia in the population of Vojvodina]. / Ispitivanje asocijacije HLA antigena i leukemija u populaciji Vojvodine.

INTRODUCTION: One of the most fascinating areas of research within the field of histocompatibility at present time concerns an observation that a major human histocompatibility system, HLA, is deeply involved in development of a great number of diseases. MATERIALS AND METHODS: HLA class I antigens were investigated in 225 cases with various kinds of leukemia: 112 patients with acute myeloblastic leukemia (AML), 31 with acute lymphoblastic leukemia (ALL), 44 with chronic myelocytic leukemia (CML) and 38 with chronic lymphocytic leukemia (CLL). Applying method of microlymphocytotoxicity, 13, 19 and 8 antisera were used for A, B and C loci respectively. Control group comprised 300 unrelated persons, whose phenotypic frequencies were used to calculate the relative risk (RR), while for values of RR greater than 1, we calculated etiologic fraction (EF), and for negative RR values we calculated preventive fraction (PF). RESULTS: Results of investigation showed that RR in AML was: for A2 = 1.144, for A3 = 1.038, for A29 = 1.814, for A34 = 2.69, for B7 = 1.06, for B14 = 1.74, for B17 = 1.65 and for B21 = 2.49 and B35 = 1.77 with value of chi 2 test of 4.62 and 4.63; that RR in ALL was: for A1 = 1.61, for A2 = 1.1, for A10 = 1.23, for A11 = 1.57, for A30 = 1.4, for A32 = 2.2, for B7 = 2.81 with value of chi 2 test 4.39; that RR in CML was: for A2 = 1.21, for A32 = 1.89, for B7 = 1.52, for B12 = 1.2 and for B15 = 3.28 with value of chi 2 test of 5.89; and that RR in CLL was: for A1 = 1.35 with value of chi 2 test of 3.973, RR for A2 = 1.02, for A28 = 1.97, for A32 = 1.25, for B5 = 1.44, for B8 = 1.27, for B13 = 1.91 without statistically significant differences of frequencies except for A1. Investigation of differences between haplotype frequencies among controls and patients showed statistically significant difference of A10 B40 haplotype in CML with RR value 7.24, while there were no statistically significant differences between controls and other leukemias. DISCUSSION: Our results of investigation showed statistically significant differences between HLA frequencies in the control group and investigated diseases, and that the relative risk is under 1, and values of chi 2 test under borderline values for B21 and B35 in AML, for B7 in ALL, for B15 in CML and for A1 in CLL. CONCLUSION: Results of this investigation point to an association between HLA system and leukemias. This association is of great importance because it provides a new tool for investigation of genetics and etiology of abovementioned diseases.