New Oxazolo[5,4-d]pyrimidines as Potential Anticancer Agents: Their Design, Synthesis, and In Vitro Biological Activity Research.

Cancer is a large group of diseases in which the rapid proliferation of abnormal cells generally leads to metastasis to surrounding tissues or more distant ones through the lymphatic and blood vessels, making it the second leading cause of death worldwide. The main challenge in designing a modern anticancer therapy is to develop selective compounds that exploit specific molecular targets. In this work, novel oxazolo[5,4-d]pyrimidine derivatives were designed, synthesized, and evaluated in vitro for their cytotoxic activity against a panel of four human cancer cell lines (lung carcinoma: A549, breast adenocarcinoma: MCF7, metastatic colon adenocarcinoma: LoVo, primary colon adenocarcinoma: HT29), along with their P-glycoprotein-inhibitory ability and pro-apoptotic activity. These oxazolo[5,4-d]pyrimidine derivatives, which are structurally similar to nucleic purine bases in general, are characterized by the presence of a pharmacologically favorable isoxazole substituent at position 2 and aliphatic amino chains at position 7 of the condensed heterocyclic system. In silico analysis of the obtained compounds identified their potent inhibitory activity towards human vascular endothelial growth factor receptor-2 (VEGFR-2). Molecular docking was performed to assess the binding mode of new derivatives to the VEGFR-2 active site. Then, their physicochemical, pharmacokinetic, and pharmacological properties (i.e., ADME-administration, distribution, metabolism, and excretion) were also predicted to assess their druglikeness. In particular, compound 3g (with a 3-(N,N-dimethylamino)propyl substituent) was found to be the most potent against the HT29 cell line, with a 50% cytotoxic concentration (CC50) of 58.4 µM, exceeding the activity of fluorouracil (CC50 = 381.2 µM) and equaling the activity of cisplatin (CC50 = 47.2 µM), while being less toxic to healthy human cells (such as normal human dermal fibroblasts (NHDFs)) than these reference drugs. The results suggest that compound 3g is a potentially promising candidate for the treatment of primary colorectal cancer.

Local Effects of a 1940 nm Thulium-Doped Fiber Laser and a 1470 nm Diode Laser on the Pulmonary Parenchyma: An Experimental Study in a Pig Model.

The lungs are a common site of metastases from malignant tumors. Their removal with a minimal but safe tissue margin is essential for the long-term survival of patients. The aim of this study was to evaluate the usefulness of a 1940 nm thulium-doped fiber laser (TDFL) and a 1470 nm diode laser (DL) in a pig model of lung surgery that involved the incision and excision of lung tissue. Histopathological analysis was performed on days 0 and 7 after surgery. Neither TDFL nor DL caused significant perioperative or postoperative bleeding. Histological analysis revealed the presence of carbonized necrotic tissue, mixed fibrin-cellular exudate in the superficial zone of thermal damage and bands of deeper thermal changes. The mean total width of thermal damage on day 0 was 499.46 ± 61.44 and 937.39 ± 109.65 µm for TDFL and DL, respectively. On day 7, cell activation and repair processes were visible. The total width of thermal damage was 2615.74 ± 487.17 µm for TDFL vs. 6500.34 ±1118.02 µm for DL. The superficial zone of thermal damage was narrower for TDFL on both days 0 and 7. The results confirm the effectiveness of both types of laser in cutting and providing hemostasis in the lungs. TDFL caused less thermal damage to the lung parenchyma than DL.

Usefulness of Thulium-Doped Fiber Laser and Diode Laser in Zero Ischemia Kidney Surgery-Comparative Study in Pig Model.

BACKGROUND: The aim of this study was to evaluate the usefulness of a thulium-doped fiber laser and a diode laser in zero ischemia kidney surgery, by carrying out a comparative study in a pig model. MATERIAL AND METHODS: Research was carried out on 12 pigs weighing 30 kg each. A thulium-doped fiber laser (TDFL) and a diode laser (DL) operating at wavelengths of 1940 and 1470 nm, respectively, were used. The cut sites were assessed both macroscopically and microscopically. The zone of thermal damage visible in the histopathological preparations was divided into superficial and total areas. RESULTS: During partial nephrectomy, moderate to minimal bleeding was observed, which did not require additional hemostatic measures. All animals survived the procedure. On day 0, the total thermal damage depth was 837.8 µm for the TDFL and 1175.0 µm for the DL. On day 7, the depths were 1556.2 and 2301.7 µm, respectively. On day 14, the overall thermal damage depth for the DL was the greatest (6800 µm). The width of the superficial zone was significantly reduced on days 7 and 14 after TDFL application. CONCLUSION: Both lasers are suitable for partial wedge nephrectomy without ischemia in pigs. The TDFL produced similar or better hemostasis than the DL, with a smaller zone of thermal damage and, therefore, seems more suitable for application in human medicine.

Study of Flebogrif®-A New Tool for Mechanical Sclerotherapy-Effectiveness Assessment Based on Animal Model.

Sclerotherapy is the chemical occlusion of vessels using an intravenous injection of a liquid or foamed sclerosing agent that is used in the therapy of blood and lymphatic vessels malformations in the young, and for spider veins, smaller varicose veins, hemorrhoids and hydroceles in adults. This study aimed to assess the effectiveness of mechanosclerotherapy of venous veins with a new device-Flebogrif®-based on an animal model. The experiment was performed on nine Polish Merino sheep weighing 40-50 kilograms. The animals were anesthetized intravenously. The material was divided into three groups: two experimental (1 and 2) and control (3) group. The first experimental group was treated with the use of Flebogrif® and a sclerosant simultaneously, while only Flebogrif® was used in the second experimental group. Flebogrif® was applied into the lateral saphenous vein of both pelvic limbs. The vessel wall thickness was estimated at four points of the histological image in mm (V1, V2, V3, V4). For one month, the animals were euthanized, and the occlusion rate of the treated veins and changes in the vein wall were determined. Histological slides were analyzed under a light microscope and histometry of the vein wall was performed. The Shapiro-Wilk test and the quantity of the investigated parameter groups allowed for using a non-parametric method at four points to compare thickness measurements (the Mann-Whitney test), with p < 0.05. The Mann-Whitney test indicated statistically significant differences between both experimental groups. The results obtained from morphometrical and histological analysis showed better results in the first experimental group than those of the second experimental group. Finally, statistical analysis revealed significant differences between the both the experimental group and control group in morphological analysis. The achieved results allowed us to conclude that the simultaneous use of Flebogrif® and a sclerosant yielded better results of vein lumen reduction than the use of Flebogrif® alone.

Comparison of A 1940 nm Thulium-Doped Fiber Laser and A 1470 nm Diode Laser for Cutting Efficacy and Hemostasis in A Pig Model of Spleen Surgery.

Partial and total splenectomies are associated with a high risk of substantial blood loss. Lasers operating at wavelengths strongly absorbed by water have the potential to improve hemostasis and cut while providing a narrow zone of thermal damage. The aim of this study is to compare a thulium-doped fiber laser (TDFL) emitting a wavelength of 1940 nm and a diode laser (DL) operating at 1470 nm for spleen surgery in a pig model. A partial splenectomy and spleen incisions were made in 12 animals using the two laser devices. The hemostasis was evaluated visually during surgeries. Post-mortem and histopathological evaluations were done on days 0, 7, and 14 following surgery. Neither TDFL nor DL caused bleeding on day 0 or delayed bleeding. On day 14, pale streaks at the site of incision were slightly wider after cutting with DL than with TDFL. Histological analysis revealed a carbonized zone with exudation and a deeper zone of thermal tissue damage on day 0. The width of the thermal changes was 655.26 ± 107.70 µm for TDFL and 1413.37 ± 111.85 µm for DL. On day 7, a proliferation of fibroblasts and splenocytes was visible, as well as a formation of multinucleated giant cells adjacent to the residues of carbonization. The zone of thermal damage was broader for DL (1157.5 ± 262.77 µm) than for TDFL (682.22 ± 116.58 µm). On day 14, cutting sites were filled with connective and granulation tissues with the residues of carbonization. The zone of thermal damage was narrower for TDFL (761.65 ± 34.3 µm) than for DL (1609.82 ± 202.22 µm). Thus, both lasers are efficient in spleen surgery, providing good hemostasis. However, TDFL produces a narrower zone of thermal damage, which suggests its better efficiency for spleen surgery, especially when performing more precise procedures.

Low-level light therapy reduces platelet destruction during extracorporeal circulation.

Extracorporeal circulation causes many deleterious effects on blood cells. Low-level light therapy (LLLT) in the red/near-infrared spectral range is known for its cytoprotective properties but its use during cardiopulmonary bypass (CPB) has not yet been studied. We aimed to assess whether LLLT protects platelets during CPB. 24 pigs were connected to 1-hour-CPB and observed for the next 23 hours. In 12 animals, blood circulating through the oxygenator was treated with LLLT. Platelet count and function were monitored throughout the experiment. The decrease in platelet count was greater in the control group, especially during CPB and after 24 hours. In LLLT group CD62P expression remained quite stable up to the 12th hour of the experiment, whereas in the control group it continuously decreased till the end of observation. Platelets in the control group were more prone to aggregation in the postoperative period than at the beginning of the experiment, whereas platelets in the LLLT group aggregated similarly or less intense. Limitation of platelet loss, pattern of aggregation and CD62P expression suggest that LLLT may stabilize platelet function during CPB and diminish the negative effects associated with the interaction of cells with an artificial surface.

Preliminary Evaluation of Thulium Doped Fiber Laser in Pig Model of Liver Surgery.

Partial liver resection is a treatment of choice for liver tumors; the range of parenchyma excision varies from a small part of the tissue surrounding the neoplasm up to 70% of the organ. One of the major concerns during liver resection is blood loss. Thulium lasers which are characterized by the length of emission wave corresponding to a peak absorption of water create a new possibility of cutting tissues efficiently with minimal thermal damage and concurrently providing a good hemostasis control. The aim of our study was to evaluate an impact of liver transection with thulium doped fiber laser on an intraoperative bleeding and histopathological changes during postoperative period in swine model. Ten animals were subjected to open surgery partial liver resection and an incision of liver tissue with an all-fiber, diode-pumped, and continuous-wave Tm3+-doped fiber laser emitting 37.4 W of output power at ~1.94 µm wavelength. The macroscopic and histopathological evaluation was performed intraoperatively as well as 7 and 14 days after surgery. Macroscopically almost no bleeding was observed during surgery and no signs of bleeding were stated after 7 and 14 days. Histopathological analysis of the transection margin revealed a thermal damage area ranging in depth from 620.23 ± 23.82 µm on the day of surgery to 1817.70 ± 211.98 µm after 7 days. In the samples taken intraoperatively and after 7 days a superficial zone of carbonization was visibly separated from the deeper changes. After 14 days one 765.35 µm deep zone characterized by a granulation was present. In conclusion, thulium doped fiber laser is efficacious in cutting with a narrow zone of thermal injury and provides a good hemostasis during liver transection, thus being a potential tool for oncotic liver surgery.

Low-Level Light Therapy Protects Red Blood Cells Against Oxidative Stress and Hemolysis During Extracorporeal Circulation.

Aim: An activation of non-specific inflammatory response, coagulation disorder, and blood morphotic elements damage are the main side effects of the extracorporeal circulation (ECC). Red-to-near-infrared radiation (R/NIR) is thought to be capable of stabilizing red blood cell (RBC) membrane through increasing its resistance to destructive factors. We focused on the development of a method using low-level light therapy (LLLT) in the spectral range of R/NIR which could reduce blood trauma caused by the heart-lung machine during surgery. Methods: R/NIR emitter was adjusted in terms of geometry and optics to ECC circuit. The method of extracorporeal blood photobiomodulation was tested during in vivo experiments in an animal, porcine model (1 h of ECC plus 23 h of animal observation). A total of 24 sows weighing 90-100 kg were divided into two equal groups: control one and LLLT. Blood samples were taken during the experiment to determine changes in blood morphology [RBC and white blood cell (WBC) counts, hemoglobin (Hgb)], indicators of hemolysis [plasma-free hemoglobin (PFHgb), serum bilirubin concentration, serum lactate dehydrogenase (LDH) activity], and oxidative stress markers [thiobarbituric acid reactive substances (TBARS) concentration, total antioxidant capacity (TAC)]. Results: In the control group, a rapid systemic decrease in WBC count during ECC was accompanied by a significant increase in RBC membrane lipids peroxidation, while in the LLLT group the number of WBC and TBARS concentration both remained relatively constant, indicating limitation of the inflammatory process. These results were consistent with the change in the hemolysis markers like PFHgb, LDH, and serum bilirubin concentration, which were significantly reduced in LLLT group. No differences in TAC, RBC count, and Hgb concentration were detected. Conclusion: We presented the applicability of the LLLT with R/NIR radiation to blood trauma reduction during ECC.

[Minimally invasive access to the abdominal aorta in pigs]. / Malo inwazyjne dojscie operacyjne do aorty brzusznej u swin.

Experiments were conducted on 10 sows weighing 50-60 kg. Animals were randomly divided into two experimental groups comprising 5 individuals each. Surgical procedure of reaching the abdominal aorta below the renal arteries was performed using two different methods. In animals from the first group the procedure involved cutting the linea alba in order to obtain access to the abdominal aorta through the abdominal cavity. In the second group the access to the abdominal aorta was obtained by retroperitoneal approach. The approach was made in lower lateral one-third of the abdomen cutting through the muscles and without opening the peritoneal cavity. The study showed superiority of the second surgical method over the median approach along the linea alba in the form of: better animal survival (100% of animals), shorter procedure time, lack of postoperative complications, shorter recovery period and better general postoperative condition