Antiproliferative Properties of Triterpenoids by ECIS Method-A New Promising Approach in Anticancer Studies?

Electric cell-substrate impedance sensing is an advanced in vitro impedance measuring system which uses alternating current to determine behavior of cells in physiological conditions. In this study, we used the abovementioned method for checking the anticancer activities of betulin and betulinic acid, which are some of the most commonly found triterpenes in nature. In our experiment, the threshold concentrations of betulin required to elicit antiproliferative effects, verified by MTT and LDH release methods, were 7.8 µM for breast cancer (T47D), 9.5 µM for lung carcinoma (A549), and 21.3 µM for normal epithelial cells (Vero). The ECIS results revealed the great potential of betulin and betulinic acid's antitumor properties and their maintenance of cytotoxic substances to the breast cancer T47D line. Moreover, both substances showed a negligible toxic effect on healthy epithelial cells (Vero). Our investigation showed that the ECIS method is a proper alternative to the currently used assay for testing in vitro anticancer activity of compounds, and that it should thus be introduced in cellular routine research. It is also a valuable tool for live-monitoring changes in the morphology and physiology of cells, which translates into the accurate development of anticancer therapies.

Osteoarthritis of the knee - biochemical aspect of applied therapies: a review.

The most prevalent form of arthritis is osteoarthritis (OA) of the knee, which is characterized by a degeneration of articular cartilage resulting in the development of osteophytes, or bone spurs. Main goals of OA treatment are to reduce pain, slow the disease progression, and improve joint function and the quality of life. The purpose of this study was to verify all the therapies recommended by the European Society for Clinical and Economic Aspects of Osteoporosis and Osteoarthritis (ESCEO) from the biochemical point of view. Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit the synthesis of eicosanoids, whereas paracetamol prevents the production of prostaglandin (PG) by interacting with peroxidase (POX) site of the prostaglandin H2 synthase complex. Tramadol is an opioid that has a dual mechanism of action: it binds to the µ-opioid receptor and it inhibits serotonin and adrenaline. Corticosteroids, which are also prescribed for OA pain, inhibit the activity of phospholipase A2 and block the synthesis of arachidonate-derived eicosanoids. Symptomatic slow-acting drugs for osteoarthritis (SYSADOA) are drugs that are well tolerated by patients and help to restore proteoglycan matrix of the cartilage. These drugs include compounds that naturally build articular cartilage. The articular cartilage, as well as the bone located around the cartilage, are destroyed as osteoarthritis progresses. Thus, bisphosphonates, commonly used in the treatment of osteoporosis, were evaluated as potential therapy. However, there is no official recommendation for their use in therapy. The aim of the study was to analyze the biochemical mechanisms of principal drugs used for the treatment of knee OA. Therefore, a narrative review summarizing the current knowledge regarding the applied therapies was prepared.

Intense remodeling of extracellular matrix within the varicose vein: the role of gelatinases and vascular endothelial growth factor.

BACKGROUND: Increased blood pressure in the varicose veins (VV) can contribute to the overexpression of matrix metalloproteinases (MMPs), affecting the endothelium, smooth muscle, and extracellular matrix of the vein wall. Gelatinases (MMP-2 and MMP-9), hypoxia, and inflammation occurring in the VV wall contribute to the increased expression of vascular endothelial growth factor (VEGF). AIMS: Our objective was to analyze the concentration of gelatinases and VEGF in the great saphenous VV wall and plasma of patients. METHODS: In total, 65 patients (2nd degree according to clinical state classification, etiology, anatomy, and pathophysiology-CEAP classification) aged 22 to 70 were enrolled. Control veins (n = 10) were collected from the patients who underwent coronary artery bypass graft surgery. Control plasma (n = 20) was obtained from healthy individuals. Gelatinases and VEGF levels were measured with the usage of ELISA method. RESULTS: A significant increase in MMP-9 (11.2 vs. 9.98 ng/mg of protein) and VEGF (41.06 vs. 26 ng/g of protein) concentration in VV wall compared with control veins was observed. A positive correlation between VEGF versus MMP-2 (p = 0.03, r = 0.27) was found in the VV wall. However, no correlation was found between the concentration of VEGF and MMP-9 (p = 0.4, r = 0.11) in the VV wall. In addition, no statistical differences between MMP-9, MMP-2, and VEGF levels in plasma of VV patients compared with controls were noticed. CONCLUSIONS: The results of the present study confirm that VV's patients have altered expression of MMPs and VEGF. Overexpression of MMP-9 and VEGF in the VV wall may contribute to the spreading of inflammatory process and suggests the intense remodeling of extracellular tissue within the VV wall.

THE EVOLUTION OF TREATMENT AT THE ORTHOPEDICS AND REHABILITATION CLINIC.

OBJECTIVE: Introduction: The last 20 years have been a period of huge changes in Polish healthcare, both in terms of medical progress as well as organization and financing. These changes, and especially the newly introduced queuing systems significantly influenced the changes in the profile of hospital admission. The aim: To analyze changes in the patient profile, causes and time of hospitalization, as well as the waiting time for hospitalization, in 1996-2019 Lublin SPSK4 Orthopedics and Rehabilitation Clinic. PATIENTS AND METHODS: Material and methods: Patients' medical records were analyzed, including qualification cards. The waiting time for admission to the hospital, the time of hospitalization, the cause of hospitalization, gender and the patient's place of residence were analyzed. RESULTS: Results and conclusions: 1. During the 13 years analyzed, the number of hospitalizations increased from 452 to 1387 patients a year. 2. The waiting time for hospitalization increased from an average of 2 months in 1996 to even 2 years (in the case of patients with chronic category) in 2020. 3. The average length of hospitalization changed compared to the level of 1996 (8 weeks) in the case of patients with the early neurological category increased slightly (9 weeks), in the case of other patients - it decreased to 4 weeks, respectively - chronically ill patients, 3 weeks - patients from the systemic and weekly rehabilitation category - orthopedic patients. 4. The scope of the reasons for hospitalization, but also methods of treatment has expanded significantly. 5. Patient demographic profile in the examined aspects (origin, gender) - remained similar.

FAT TISSUE AND SYNTHESIZED BY HER ADIPOKINES AS MARKERS INDICATING THE DEVELOPMENT AND PROGRESS OF OSTEOARTHRITIS.

Arthrosis as osteoarthritis is a global problem that affects more and more people and is associated with severe chronic pain, reduced mobility and, consequently, disability. The etiology of degenerative disease is complex and depends on many factors. However, its course was not fully understood. One of the factors affecting the development of arthrosis is obesity. Obesity is a growing problem. Over the past 30 years, the number of overweight people has almost doubled. In people suffering from obesity, whose body mass index is above 30kg/m2, the risk of developing degenerative changes in articular cartilage is six times higher than the risk of developing this disease in people with normal body weight. Osteoarthritis is detected when the symptoms get worse where the changes are already at some stage. Therefore, a lot of research is currently underway to find suitable biomarkers, which would indicate the potential development of degenerative changes in the future and at the same time the possibility of inhibiting their activity. One of them may be adipokines, which are synthesized by adipose tissue and affect cartilage. In obese people, adipokines may contribute to the inflammation of the low charterer, whichaccompanies both obesity and arthrosis. These compounds can be specific biomarkers to assess the degree of progression and severity of osteoarthritis. The aim: To assess the importance of obesity and adipokines produced by adipose tissue as specific markers of arthrosis.

Decreased nitric oxide serum level after pituitary adenoma resection.

BACKGROUND: Nitric oxide (NO) is synthesized by the conversion of Arginine (Arg) into the NO and Citrulline (Cit). Although the NO is involved into the pathogenesis of several physiological and pathological processes, the role of NO in pituitary adenomas (PA) progression is not determined. Our purpose was to evaluate the relationship between NO and PA as well as the effect of tumor resection on NO metabolites level in serum. METHODS: The study group consisted of 21 patients with PA, 18 patients with macroadenomas and 3 with microadenomas. Venous blood samples were collected at two time-points; 1) before the surgery and 2) 3-5 days after PA resection. Arg and Cit levels were determined by the automated ion-exchange chromatography with usage of Amino Acids Analyser (AAA 400). Commercially available kit for the evaluation of nitrate/nitrite serum levels was applied for indirect assessment of serum NO level. RESULTS: Significant decrease in NO concentration after the surgery was observed in comparison with the time-point 1. Arg level did not significantly change during the study period. Cit level was ranged below the detection limit of applied method. CONCLUSIONS: The decrease of NO level after the pituitary adenoma resection indicates the relationship between NO synthesis and PA occurrence.

Antioxidative mechanism in the course of varicose veins.

Objective Our objective was to evaluate the state of oxidative stress in the great saphenous varicose vein wall and blood of varicose vein patients taken from the antecubital vein. Methods The superoxide dismutase, reduced glutathione (GSH) and total antioxidant status were measured with commercially available colorimetric kits in erythrocytes, plasma and varicose vein wall of 65 patients (second degree of clinical state classification, etiology, anatomy and pathophysiology) aged 22-70 (49 women, 16 men) in comparison to normal great saphenous vein walls collected from 10 patients who underwent coronary artery bypass graft and blood collected from 20 healthy individuals. Results A statistically significant decrease (p < 0.001) in superoxide dismutase activity in erythrocytes and the increase (p < 0.05) in superoxide dismutase activity in varicose vein has been observed. There have been no significant changes in the concentration of GSH in plasma and in varicose vein. The decreased concentration of total antioxidant status in plasma (p < 0.001) and in varicose vein wall (p < 0.05) in comparison to the control has been noticed. Conclusion The varicose vein patients are affected by oxidative stress. Our results indicate impaired antioxidant defense mechanism in the blood of varicose vein patients. In contrast to the blood, an increased process of antioxidant defense in the varicose vein wall was noticed.

Different patterns of gelatinolytic activity in pituitary macro- and microadenomas.

OBJECTIVE: Gelatinases, Matrix MetalloProteinase(MMP)-2 and MMP-9, belong to zinc-dependent endopeptidases involved in several physiological and pathological processes including inflammation and tumor development. Because the information about the involvement of gelatinases in pituitary adenoma (PA) development are scant, our objective was the analysis of MMP-2 and MMP-9 activity in serum and tumor tissue of PA patients. PATIENTS AND METHODS: Twenty one patients with PA (macroadenoma n=18, microadenoma n=3), qualified to the endoscopic resection of tumors were enrolled. Venous blood samples were collected before the surgery and PA tissue was collected during the surgery. Tissue material was homogenized in a buffer containing 0.1M Tris-HCl pH 7.4 and centrifuged. The supernatant was set to the equal protein content 18µg/sample. Protein level in tissue samples was estimated with Bradford method. MMP-2 and MMP-9 analysis in serum and tissue was performed with gelatin zymography. RESULTS: The proteolytically activated forms of MMPs were not observed in the analyzed sera. Serum activities of MMP-2 and MMP-9 did not statistically differ between patients with micro and macroadenomas. The analysis of material obtained from tissue of microadenomas showed slightly lower activities of both forms of MMP-9 (pro-MMP-9 and MMP-9/lipokalin heterodimer). Simultaneously the increased activity of pro-MMP-2 in comparison to macroadenomas was observed. Although differences observed did not reach statistical significance, only in the case of microadenomas the presence of the active form of MMP-2 (molecular weight 65kDa band) was observed. CONCLUSION: In the course of PA growth the change the biochemical profile of the gelatinolytic activity within the tumor tissue is observed. Initially, the higher activity of MMP-2 in microadenomas and elevated activity of MMP-9 in macroadenomas were detected.

Actylise treatment does not influence nitric oxide metabolites serum level.

BACKGROUND: Nitric oxide (NO) is synthesized by Nitric Oxide Synthases (NOS), the family of enzymes capable to conduct the conversion of Arginine (Arg) into the NO and Citrulline (Cit). Currently, only the administration of recombinant tissue plasminogen activator (rtPA) is recommended for acute ischemic stroke (AIS) treatment. To allow solubility of rtPA, Arg is added as a constituent of the drug. Our purpose was to check the effect of alteplase administration on NO metabolites concentration in the blood. METHODS: Eighteen AIS patients were selected into the study. Nine of them received thrombolytic therapy (rtPA group). The serum samples were obtained at 3 time-points for rtPA group (time-point 0: 1st-4th hour of stroke; time-point 1: immediately after rtPA administration; time-point 3: on day 5-7 from stroke onset). Remaining patients (non-rtPA group) had blood collection at two time-points: time-point 1: 1st-10th hour of stroke and time-point 2: on day 5-7 of stroke. Arg and Cit were determined by the automated ion-exchange chromatography using Amino Acids Analyzer. NO serum level was indirectly evaluated with the usage of commercially available kits that measuring the nitrate/nitrite level. RESULTS: Significant increase of Arg serum level was noticed at time-point 1, directly after the iv thrombolysis in comparison to non-rtPA group. However, the products of the reaction catalyzed by NOS (NO and Cit) did not rise after the thrombolysis. CONCLUSIONS: Current study showed that Arg administration simultaneously with rtPA, as a constituent of Actylise, does not affect serum NO metabolites level.

Simvastatin Effect on Calcium and Silicon Plasma Levels in Postmenopausal Women with Osteoarthritis.

Postmenopausal women more often suffered from knee osteoarthritis and its pathogenesis still remains unclear. Calcium and silicon are significant elements involved in bone and joint metabolism, especially in older people. Cardiovascular diseases are common worldwide and simvastatin is the most prescribed drug in such population of patients. The purpose of this study was to evaluate the effect of simvastatin administration on calcium and silicon concentration in the plasma of postmenopausal women with osteoarthritis. Sixty postmenopausal mild hypercholesterolemic women (mean age 61.4 years, range 54-68) were enrolled. Thirty patients received simvastatin (20 or 40 mg/day) for at least 1 year before being enrolled (simvastatin "+" group). Control group consists of remaining 30 women (simvastatin "-"group). Silicon and calcium concentrations were measured spectrophotometrically. Plasma simvastatin level was determined 3 h after the drug administration using HPLC-UV-Vis. Calcium but not silicon level was significantly lower in patients receiving simvastatin in comparison with non-statin group (1.91 ± 0.32 vs. 2.33 ± 0.19 mmol/l, p < 0.05). A weak but significant positive correlation between plasma silicon and simvastatin levels (r = 0.3, p < 0.05) was observed; this may be due to the fact that simvastatin contains silicon dioxide as an inactive ingredient. The mean simvastatin concentration was 9.02 ng/ml. All hypotheses were verified at the significance level of p < 0.05. A statistically significant decrease in the plasma calcium concentration of postmenopausal women, treated with simvastatin suggests that simvastatin may play a role in calcium metabolism in postmenopausal women with osteoarthritis. Positive correlation of simvastatin concentration with silicon level in the plasma suggests that both might prompt the positive effect of osteoarthritis treatment.

Characteristics of hematopoietic stem cells of umbilical cord blood.

Umbilical cord blood collected from the postpartum placenta and cord is a rich source of hematopoietic stem cells (HSCs) and is an alternative to bone marrow transplantation. In this review we wanted to describe the differences (in phenotype, cytokine production, quantity and quality of cells) between stem cells from umbilical cord blood, bone marrow and peripheral blood. HSCs present in cord blood are more primitive than their counterparts in bone marrow or peripheral blood, and have several advantages including high proliferation. With using proper cytokine combination, HSCs can be effectively developed into different cell lines. This process is used in medicine, especially in hematology.

Antioxidants in patients receiving total parenteral nutrition after gastrointestinal cancer surgery.

Total parenteral nutrition (TPN) is essential for patients with postoperative impairing gastrointestinal function who are unable to receive and absorb oral/enteral feeding for at least 7 days. Oxidative stress plays a major role in the ethiopathogenesis of cancers. In this study, total antioxidant status (TAS), glutathione peroxidase (GPx), superoxide dismutase, malondialdehyde and ascorbic acid were studied in patients operated because of small intestine, colorectal or pancreatic cancer and subsequently receiving TPN in comparison with patients receiving standard nutrition after the operation. TAS level and GPx activity were decreased in patients with small intestine cancer but did not differ in patients with colorectal and pancreatic cancer before and after surgery. In all patient groups receiving TPN, superoxide dismutase activity after the surgery was kept at the same level as before. On the fifth day after the surgery, malondialdehyde concentration in each group was restored to the value observed before surgery. On the fifth day of TPN treatment, ascorbic acid concentration was increased in every group of patients. TPN applied during the postoperative period alleviates oxidative stress resulting from surgery. In the case of small intestine cancer, the addition of vitamins and antioxidants to the nutrition mixture seems to result in depletion of antioxidant enzymes' activities.