The development of new methodology for determination of vincristine (VCR) in human serum using LC-MS/MS-based method for medical diagnostics

In this article, we have presented the development and validation of a rapid and sensitive reversed-phase liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the determination of vincristine (VCR) in patient serum samples. Chromatographic separation was achieved on a Kinetex(®) (Singapore) column using a mobile phase consisting of 25 mM acetic acid and 0.3% formic acid (A) and methanol (B) in a gradient elution mode at a flow rate of 0.3 mL/min. The VCR and internal standard (vinblastine) were monitored using the multiple reaction monitoring mode under positive electrospray ionization. The lower limit of quantification (LLOQ) was 0.67 ng/mL, and the upper limit of quantification (ULOQ) was 250 ng/mL for VCR. The calculated values of LOD and LOQ for VCR were 0.075 and 0.228 ng/mL, respectively. The calibration curve was linear over the VCR concentration range of 1.0–250 ng/mL in serum. The intra- and inter-day precision and precision were within the generally accepted criteria for the bioanalytical method (<15%). The method was successfully applied to the analysis of serum samples in clinical practice

The correlation of mutations and expressions of genes within the PI3K/Akt/mTOR pathway in breast cancer : a preliminary study

There is an urgent need to seek new molecular biomarkers helpful in diagnosing and treating breast cancer. In this elaboration, we performed a molecular analysis of mutations and expression of genes within the PI3K/Akt/mTOR pathway in patients with ductal breast cancer of various malignancy levels. We recognized significant correlations between the expression levels of the studied genes. We also performed a bioinformatics analysis of the data available on the international database TCGA and compared them with our own research. Studies on mutations and expression of genes were conducted using High-Resolution Melt PCR (HRM-PCR), Allele-Specific-quantitative PCR (ASP-qPCR), Real-Time PCR molecular methods in a group of women with ductal breast cancer. Bioinformatics analysis was carried out using web source Ualcan and bc-GenExMiner. In the studied group of women, it was observed that the prevalence of mutations in the studied PIK3CA and AKT1 genes was 29.63%. It was stated that the average expression level of the PIK3CA, PIK3R1, PTEN genes in the group of breast cancer patients is lower in comparison to the control group, while the average expression level of the AKT1 and mTOR genes in the studied group was higher in comparison to the control group. It was also indicated that in the group of patients with mutations in the area of the PIK3CA and AKT1 genes, the PIK3CA gene expression level is statistically significantly lower than in the group without mutations. According to our knowledge, we demonstrate, for the first time, that there is a very strong positive correlation between the levels of AKT1 and mTOR gene expression in the case of patients with mutations and without mutations

Advances in Management of Bladder Cancer&mdash;The Role of Photodynamic Therapy

Photodynamic therapy (PDT) is a non-invasive and modern form of therapy. It is used in the treatment of non-oncological diseases and more and more often in the treatment of various types of neoplasms in various locations including bladder cancer. The PDT method consists of local or systemic application of a photosensitizer, i.e., a photosensitive compound that accumulates in pathological tissue. Light of appropriate wavelength is absorbed by the photosensitizer molecules, which in turn transfers energy to oxygen or initiates radical processes that leads to selective destruction of diseased cells. The technique enables the selective destruction of malignant cells, as the photocytotoxicity reactions induced by the photosensitizer take place strictly within the pathological tissue. PDT is known to be well tolerated in a clinical setting in patients. In cited papers herein no new safety issues were identified. The development of anti-cancer PDT therapies has greatly accelerated over the last decade. There was no evidence of increased or cumulative toxic effects with each PDT treatment. Many modifications have been made to enhance the effects. Clinically, bladder cancer remains one of the deadliest urological diseases of the urinary system. The subject of this review is the anti-cancer use of PDT, its benefits and possible modifications that may lead to more effective treatments for bladder cancer. Bladder cancer, if localized, would seem to be a good candidate for PDT therapy since this does not involve the toxicity of systemic chemotherapy and can spare normal tissues from damage if properly carried out. It is clear that PDT deserves more investment in clinical research, especially for plant-based photosensitizers. Natural PS isolated from plants and other biological sources can be considered a green approach to PDT in cancer therapy. Currently, PDT is widely used in the treatment of skin cancer, but numerous studies show the advantages of related therapeutic strategies that can help eliminate various types of cancer, including bladder cancer. PDT for bladder cancer in which photosensitizer is locally activated and generates cytotoxic reactive oxygen species and causing cell death, is a modern treatment. Moreover, PDT is an innovative technique in oncologic urology

Natural and Synthetic Coumarins and their Pharmacological Activity

Coumarins are a structurally diverse group of natural substances derives from plants that display a host of bioactivities. In this paper, we will introduce the reader to coumarins and their applications as medicinal substances. The great diversity in courmarin structure will be discussed along with their extensive use as pharmaceutical agents. Coumarins display a wide range of antimicrobial activity and applications of coumarins as antifungal and antiviral agents will be addressed. Other properties of coumarins such as their role in neuroprotection, anticancer, and as antioxidants will also be reviewed

Combined effect of flavonoid compounds and cytostatics in cancer treatment

Aim. The aim of the study was to review the literature on the combination of cytostatics with flavonoids as a promising way to improve the cancer therapy. Material and methods. A review of Polish and foreign literature was performed. The following databases were searched: PubMed, Scopus, Science Direct, and Polish Medical Bibliography. Literature analysis. Effective strategies to inhibit the progression of cancer are needed. Compounds of natural origin, including plant polyphenols, are a part of our diet. Due to their availability, and antioxidant properties, they may serve as efficacious adjuvants in cancer therapy, enhancing the effectiveness of chemotherapeutics. Epidemiological studies have shown an inverse relationship between diets rich in fruits, vegetables, and supplements, and the risk of all causes of death from cancer. Based on their diverse biological activity, flavonoids may be potential adjuvant therapeutic agents that act synergistically with cytostatics for treatment of many types of cancer. This review of the results is a summary the research on anticancer activity of flavonoids and may also raise consciousness of consumers, who will be able to compose their diet armed with the knowledge of preventive and therapeutic anticancer properties of food ingredients. There is need for further research on polyphenols of plant origin, including interactions among food components that coexist. Another important aspect is to understand how the activity of phytocompounds depends on concentration and the presence of additional factors (e.g. microflore, metal ions), which could possibly make a compound harmful, instead of having positive theraputics effect. Elucidation of the mechanisms involved in biological activity of the described phytocompounds is essential for a better understanding of their influence on an organism

Neural tube defects: risk factors and prevention

Neural tube defects are abnormalities that can occur in the brain (anencephaly, encephalocele), spine (spina bifida, myelomeningocele, myelodysplasia), both brain and spine (craniorachischisis) or spinal column of a developing embryo that are present at birth. They arise when the neural tube, the embryonic precursor of the brain and spinal cord, fails to close during neurulation. Many cases of neural tube defects occur worldwide each year in more than 300,000 newborn babies and are a significant cause of infant death and lifelong disability. Most neural tube defects are preventable. The prevalence of these abnormalities has decreased in the past 20 to 30 years due to periconceptional folate supplementation, food fortification and decreased exposure to environmental factors. Women who are planning to conceive should be informed about the importance of folic acid in fetal development and encouraged to take 400 μg/day of folic acid supplements. Numerous research studies have shown that taking this dosage of folic acid before and during early pregnancy significantly reduces the risk of neural tube defects. For that reason it is important to increase the awareness of women in childbearing age about the necessity of primary prevention and folate intake which is a strong factor that has wide implications in public health in reducing the mortality and morbidity of offspring

The significance of NGAL and KIM-1 proteins for diagnosis of acute kidney injury (AKI) in clinical practice

Introduction. Despite advances in medical care AKI (acute kidney injury) is associated with high morbidity and mortality. The lack of adequate early renal injury biomarkers is often a problem for an early AKI diagnosis. In recent years, numerous scientific studies have been carried out which reveal new urine and serum markers to assess the period of the kidney injury before revealing its late clinical effects. In most clinical settings, AKI is due to acute renal tubular necrosis which results in protein accumulation in urine. Determination of the concentrations of proteins such as NGAL (neutrophil gelatinase-associated lipocalin) and KIM-1 (kidney injury molecule-1) are of great significance in the diagnosis of AKI. Aim. The purpose of the study was to review the literature about significance of NGAL and KIM-1 proteins for diagnosis of acute kidney injury (AKI) in clinical practice. Materials and method. Analysis of Polish and foreign literature

Imaging Flow Cytometric Analysis of Stilbene-Dependent Apoptosis in Drug Resistant Human Leukemic Cell Lines

Background: The natural compounds have been researched extensively as an alternative to the conventional chemotherapy and radiation. Stilbene derivatives appear as a group of therapeutics which deserves special attention. The present study was designed to analyze the effects of stilbene derivatives on drug resistant human leukemic cells. The aim of this work was to evaluate the apoptotic effect of stilbene derivatives in various concentrations on leukemic cells (LC) with and without resistant phenotype. Methods: Human acute promyelocytic leukemia (APL) cell lines (HL60, HL60/MX1, HL60/MX2) and acute lymphoblastic leukemia (ALL) cell lines (CEM/C1, CCRF-CEM) were studied. T-resveratrol, piceatannol, rhaponticin, deoxyrhaponticin, pterostilbene were used to stimulate apoptosis. Mitoxantrone (MIT) was applied to induce drug resistance. Results: t-Resveratrol (RES), deoxyrhaponticin (D-RHAP), rhaponticin (RHAP), pterostilbene (PTER), and piceatannol (PIC) influenced viability and induced apoptosis in all investigated cell lines. Conclusions: Our results confirmed that RES, PIC, RHAP, D-RHAP, and PTER are essential therapeutic compounds with anticancer activity exhibited by induction of apoptosis in leukemic cells with and without resistant phenotype. Stilbene-induced apoptosis in HL60/MX1, HL60/MX2, CEM/C1, and CCRF-CEM leukemia cell lines have been presented in very few studies so far and our research is an important contribution to the investigation of these substances