Phytocannabinoids CBD, CBG, and their Derivatives CBD-HQ and CBG-A Induced In Vitro Cytotoxicity in 2D and 3D Colon Cancer Cell Models.

Phytocannabinoids, compounds found in Cannabis sativa L., are used in oncology and palliative care to reduce the adverse reactions of standard therapies. Cancer patients use formulations of Cannabis sativa L. to manage the anxiety, pain, and nausea associated with cancer treatment, and there is growing evidence that some of them may exhibit anticancer properties. In this study, we tested the anticancer potential of selected cannabinoids CBD (cannabidiol) and its quinone derivative CBD-HQ (cannabidiol hydroquinone), CBG (cannabigerol) and its acid derivative CBG-A (cannabigerolic acid), as well as a combination of CBD+CBG on the colon cancer cell line SW-620. The MTT assay was used to determine the cannabinoids' ability to induce colon cancer cell death. All cannabinoids were cytotoxic at the lowest concentration (3 µg/mL). The half maximal inhibitory concentration (IC50) ranged from 3.90 to 8.24 µg/mL, depending on the substance. Cytotoxicity was confirmed in a 3D spheroidal cell culture with calcein and propidium iodide staining. The amount of intracellular reactive oxygen species (ROS) was examined using a DCF-DA assay. CBG showed the lowest antioxidant activity of all the cannabinoids tested. The level of intracellular ROS decreased only by 0.7-18%. However, CBG-A induced the strongest reduction in ROS level by 31-39%. Our results suggest that cannabinoids represent an interesting research direction with great implementation potential. These preliminary results represent the beginning of research into the potential of these substances for anticancer treatment and underscore the potential for further research.

Comparative analysis of obesity prevalence, antioxidant and oxidant status in children with Down syndrome - a sibling-controlled study.

INTRODUCTION: Down syndrome (DS), a common genetic disorder, leads to various physical, cognitive, and developmental challenges. The supplementary copy of chromosome 21 introduces an abundance of genes, which potentially can influence metabolic irregularities. The aim of the study is to conduct a comprehensive comparative assessment of oxidative stress indicators (TAS, TOS, OSI), BMI, fasting glucose, and insulin levels, HOMA-IR among children and adolescents with DS in contrast to their non-DS siblings. MATERIAL AND METHODS: and the control group (CG) comprised 20 individuals, siblings of SG (mean age 15.92 years). Anthropometric measurements were conducted. TAS, TOS, fasting glucose, and insulin were assessed. BMI, BMI SDS, OSI and HOMA-IR were calculated. RESULTS: SG vs. CG: BMI - overweight (29,19% vs. 15%), obese (19,05% vs. 5%); TAS (1.92 mmol/l vs. 1.79 mmol/l (p = 0.0015)); TOS (51.52 mmol/l vs. 33.05 mmol/l (p = 0.014)); OSI (2475.02 vs. 1949,75 (p = 0.038)); no significant differences in fasting glucose, insulin and HOMA-IR. Statistically significant correlations in SG: TOS and BMI, BMI SDS, HOMA-IR; OSI and BMI, BMI SDS, HOMA-IR; HOMA-IR and BMI SDS; fasting insulin and BMI PC; in CG: TAS and BMI; fasting glucose and fasting insulin. CONCLUSIONS: The research results indicate differences in metabolic processes between the group of individuals with DS compared to the CG, despite shared environmental conditions. The presence of an additional copy of chromosome 21 may contribute to the occurrence of metabolic disorders. These findings emphasize the need for further research that will lead to a better understanding of these relationships and contribute to the development of effective therapeutic strategies.

Predisposition to atherosclerosis in children and adults with trisomy 21: biochemical and metabolomic studies.

INTRODUCTION: Atherosclerosis, a precursor to cardiovascular disease (CVD), is deeply intertwined with lipid metabolism. The metabolic process in the Down syndrome (DS) population remain less explored. Aim of the study: This study examines the lipid profiles of DS in comparison to their siblings (CG), aiming to uncover potential atherosclerotic and CVD risks. MATERIAL AND METHODS: The study included 42 people with DS (mean age 14.17 years) and the CG - 20 individuals (mean age 15.92 years). Anthropometric measurements: BMI, BMI SDS, and TMI were calculated. Lipid profile (LP) and metabolomics were determined. RESULTS: LP: DS display significantly reduced HDL (DS vs. CG: 47±10 vs. 59 ±12 mg/dl; p = 0.0001) and elevated LDL (104 ±25 vs. 90 ±22 mg/dl; p = 0.0331). Triglycerides, APO A1, and APO B/APO A1 ratio corroborate with the elevated risk of CVD in DS. Despite no marked differences in: TCH and APO B, the DS group demonstrated a concerning BMI trend. Of 31 identified metabolites, 12 showed statistical significance (acetate, choline, creatinine, formate, glutamine, histidine, lysine, proline, pyroglutamate, threonine, tyrosine, and xanthine). However, only 8 metabolites passed the FDR validation (acetate, creatinine, formate, glutamine, lysine, proline, pyroglutamate, xanthine). CONCLUSIONS: Down syndrome individuals show distinct cardiovascular risks, with decreased HDL and increased LDL levels. Combined with metabolomic disparities and higher BMI and TMI, this suggests an increased atherosclerosis risk compared to controls.

Combination of Irinotecan and Melatonin with the Natural Compounds Wogonin and Celastrol for Colon Cancer Treatment.

Colorectal cancers are one of the leading cancers worldwide and are known for their high potential for metastasis and resistance to therapy. The aim of this study was to investigate the effect of various combination therapies of irinotecan with melatonin, wogonin, and celastrol on drug-sensitive colon cancer cells (LOVO cell line) and doxorubicin-resistant colon cancer stem-like cells (LOVO/DX cell subline). Melatonin is a hormone synthesized in the pineal gland and is responsible for circadian rhythm. Wogonin and celastrol are natural compounds previously used in traditional Chinese medicine. Selected substances have immunomodulatory properties and anti-cancer potential. First, MTT and flow cytometric annexin-V apoptosis assays were performed to determine the cytotoxic effect and the induction of apoptosis. Then, the potential to inhibit cell migration was evaluated using a scratch test, and spheroid growth was measured. The results showed important cytotoxic effects of the drug combinations on both LOVO and LOVO/DX cells. All tested substances caused an increase in the percentage of apoptotic cells in the LOVO cell line and necrotic cells in the LOVO/DX cell subline. The strongest effect on the induction of cancer cell death was observed for the combination of irinotecan with celastrol (1.25 µM) or wogonin (50 µM) and for the combination of melatonin (2000 µM) with celastrol (1.25 µM) or wogonin (50 µM). Statistically significant improvements in the effect of combined therapy were found for the irinotecan (20 µM) and celastrol (1.25 µM) combination and irinotecan (20 µM) with wogonin (25 µM) in LOVO/DX cells. Minor additive effects of combined therapy were observed in LOVO cells. Inhibition of cell migration was seen in LOVO cells for all tested compounds, while only irinotecan (20 µM) and celastrol (1.25 µM) were able to inhibit LOVO/DX cell migration. Compared with single-drug therapy, a statistically significant inhibitory effect on cell migration was found for combinations of melatonin (2000 µM) with wogonin (25 µM) in LOVO/DX cells and irinotecan (5 µM) or melatonin (2000 µM) with wogonin (25 µM) in LOVO cells. Our research shows that adding melatonin, wogonin, or celastrol to standard irinotecan therapy may potentiate the anti-cancer effects of irinotecan alone in colon cancer treatment. Celastrol seems to have the greatest supporting therapy effect, especially for the treatment of aggressive types of colon cancer, by targeting cancer stem-like cells.

miR-31-5p as a Potential Circulating Biomarker and Tracer of Clinical Improvement for Chronic Inflammatory Demyelinating Polyneuropathy.

Background: MicroRNAs are endogenous, small noncoding RNA molecules that play a pivotal role in the regulation of gene expression. MicroRNAs are involved in many biological processes such as proliferation, cell differentiation, neovascularization, and apoptosis. Studies on microRNA expression may contribute to a better understanding of the pathomechanism of chronic inflammatory demyelinating polyneuropathy (CIDP) and consequently enable the development of new therapeutic measures using antisense miRNAs (antagomirs). In this study, we evaluated the level of miR-31-5p in the serum of patients with CIDP and its correlation with the miR-31-5p level and clinical presentation and electrophysiological and biochemical parameters. Methods: The study group consisted of 48 patients, mean age 61.60 ± 11.76, who fulfilled the diagnostic criteria of a typical variant of CIDP. The expression of miR-31-5p in patient serum probes was investigated by droplet digital PCR. The results were correlated with neurophysiological findings and the patient's clinical and biochemical parameters. Results: The mean copy number of miRNA-31 in 100 µl serum was 1288.64 ± 2001.02 in the CIDP group of patients, while in the control group, it was 3743.09 ± 4026.90. There was a significant positive correlation (0.426) between IgIV treatment duration and miR-31-5p expression. Patients without IgIV treatment showed significantly lower levels of miR-31 compared to the treated group (259.44 ± 304.02 vs. 1559.48 ± 2168.45; p = 0.002). The group of patients with body weight > 80 kg showed statistically significantly lower levels of miRNA-31-5p than the patients with lower body weight (934.37 ± 1739.66 vs. 1784.62 ± 2271.62, respectively; p = 0.014). Similarly, the patients with elevated cerebrospinal fluid (CSF) protein levels had significantly higher miRNA-31-5p expression than those with normal protein levels (1393.93 ± 1932.27 vs. 987.38 ± 2364.10, respectively; p = 0.044). Conclusion: The results may support the hypothesis that miR-31-5p is strongly involved in the autoimmune process in CIDP. The positive correlation between higher miR-31-5p levels and duration of IVIg treatment may be an additional factor explaining the efficacy of prolonged IVIg therapy in CIDP.

Pediatric Population with Down Syndrome: Obesity and the Risk of Cardiovascular Disease and Their Assessment Using Omics Techniques-Review.

People with Down syndrome (PWDS) are more at risk for developing obesity, oxidative stress disorders, metabolic disorders, and lipid and carbohydrate profile disorders than the general population. The presence of an additional copy of genes on chromosome 21 (i.e., the superoxide dismutase 1 gene (SOD1) and gene coding for the cystathionine ß-synthase (CBS) enzyme) raises the risk for cardiovascular disease (CVD). As a result of disorders in metabolic processes and biochemical pathways, theoretically protective factors (low homocysteine level, high SOD1 level) do not fulfil their original functions. Overexpression of the CBS gene leads to the accumulation of homocysteine-a CVD risk factor. An excessive amount of protective SOD1, in the case of a lack of compensatory increase in the activity of catalase and peroxidase, leads to intensifying free radical processes. The occurrence of metabolic disorders and the amplified effect of oxidative stress carries higher risk of exposure of people with DS to CVD. At present, classic predispositions are known, but it is necessary to identify early risk factors in order to be able to employ CVD and obesity prophylaxis. Detailed determination of the metabolic and lipid profile may provide insight into the molecular mechanisms underlying CVD.

Targeting CD38 in Neoplasms and Non-Cancer Diseases.

CD38 is a myeloid antigen present both on the cell membrane and in the intracellular compartment of the cell. Its occurrence is often enhanced in cancer cells, thus making it a potential target in anticancer therapy. Daratumumab and isatuximab already received FDA approval, and novel agents such as MOR202, TAK079 and TNB-738 undergo clinical trials. Also, novel therapeutics such as SAR442085 aim to outrank the older antibodies against CD38. Multiple myeloma and immunoglobulin light-chain amyloidosis may be effectively treated with anti-CD38 immunotherapy. Its role in other hematological malignancies is also important concerning both diagnostic process and potential treatment in the future. Aside from the hematological malignancies, CD38 remains a potential target in gastrointestinal, neurological and pulmonary system disorders. Due to the strong interaction of CD38 with TCR and CD16 on T cells, it may also serve as the biomarker in transplant rejection in renal transplant patients. Besides, CD38 finds its role outside oncology in systemic lupus erythematosus and collagen-induced arthritis. CD38 plays an important role in viral infections, including AIDS and COVID-19. Most of the undergoing clinical trials focus on the use of anti-CD38 antibodies in the therapy of multiple myeloma, CD19- B-cell malignancies, and NK cell lymphomas. This review focuses on targeting CD38 in cancer and non-cancerous diseases using antibodies, cell-based therapies and CD38 inhibitors. We also provide a summary of current clinical trials targeting CD38.

Leukocyte-Rich Platelet-Rich Plasma as an Effective Source of Molecules That Modulate Local Immune and Inflammatory Cell Responses.

Autologous platelet-rich plasma (PRP) injection is a safe biological method used to treat various musculoskeletal diseases. By downregulation of inflammatory cytokines and stimulation of synovial fibroblasts, PRP injection is a promising adjunctive treatment for patients with chronic autoimmune inflammatory diseases such as rheumatoid arthritis. A major problem in comparing the results of clinical trials in this area is the considerable variability in the cytokine content of PRP. We presented the profile of selected growth factors and inflammatory cytokines in the obtained PRP samples and compared them with baseline serum levels to assess the efficacy of PRP as a source of those paracrine molecules. Additionally, we wanted to determine whether the difference is only quantitative, which would suggest the use of a cheaper alternative by injecting a large amount of autologous serum. For this purpose, we analyzed whole blood and PRP samples prepared using the Mini GPS III Platelet Concentration System (Biomet Inc., USA) in 31 subjects aged 35-60 years. Cellular content, seven selected growth factors, and 13 human inflammatory cytokines were evaluated. Multiplex bead immunoassays that use fluorescence-encoded beads LEGENDplex™ (BioLegend, USA) and flow cytometer measurements were used. As a result, we found a statistically significant increase in four of the growth factors tested and eight of the inflammatory cytokines tested in PRP compared to blood serum. The difference is not only quantitative but also in the composition of paracrine molecules. In conclusion, the study confirmed that PRP is an efficient source of several growth factors and some inflammatory cytokines. These data provide additional insight into the potential mechanisms of PRP's effects on cellular metabolism and inflammatory response and may contribute to a better understanding of its clinical efficacy.

The best tool for the assessment of developmental disorders in children with down syndrome: comparison of standard and specialized growth charts - cross sectional study.

Down Syndrome (DS) is a chromosomal abnormality associated with a spectrum of cognitive and physical disabilities. Children with DS are exposed to both lower and excess body weight and follow distinct growth-curve patterns that deviate significantly from those of children without chromosomal defects. Anthropometric parameters are assessed in the pediatric population with the use of growth charts. The study is based on data from 411 children and adults with DS from Poland. Detailed information concerning children and online survey results were also analyzed. Centiles and standard deviation scores (SDS) of obtained anthropometric parameters were aligned with the data using the LMS method. The study aims to identify which type of growth chart (standard vs specialized) is a leading tool for earlier detection of developmental disorders in DS. The results obtained in the two types of growth charts differed. The advantage of the specialized growth charts over the standard ones cannot be unequivocally determined. Only the combination of both tools allows to detect the development disorders early in the broadest possible way.

Anticancer and Antioxidant Activities in Ganoderma lucidum Wild Mushrooms in Poland, as Well as Their Phenolic and Triterpenoid Compounds.

The goal of this study was to the assess anti-cancer and antioxidant properties of the Ganoderma lucidum fruiting body, and to identify bioactive compounds found in their extracts. Significant antiproliferative activity was observed against MCF-7, MCF-7/DX, LOVO, LOVO/DX, MDA-MB 231, SW 620, and NHDF cell lines. With IC50 values of 25.38 µg/mL and 47.90 µg/mL, respectively, the extract was most effective against MDA-MB 231 and SW 620 cell lines. The bioactive compounds were identified using an ACQUITY UPLC-PDA-MS system. The extracts contained 13 triterpenoids and 28 polyphenols from the flavonols, phenolic acids, flavones, flavan-3-ols, and stilbenes families. Ganoderic acid derivative was found to be the most abundant triterpenoid (162.4 mg/g DW), followed by ganoderic acid B (145.6 mg/g DW). Resveratrol was the most abundant phenolic in the extract (5155.7 mg/100 g DM). The findings could explain why G. lucidum extracts are used in folk medicine.

Effectiveness of Lateral Elbow Tendinopathy Treatment Depends on the Content of Biologically Active Compounds in Autologous Platelet-Rich Plasma.

Autologous platelet-rich plasma (PRP) injection is an alternative treatment option for patients with lateral elbow tendinopathy. The treatment is supposed to accelerate tissue regeneration by providing high concentrations of growth factors derived from platelets. The aim of the study was to assess the relationship between the content of biologically active compounds in PRP and the clinical effect of the treatment. Thirty patients with lateral elbow tendinopathy treated with a single PRP injection, were evaluated. The pain intensity (measured by a visual analogue scale (VAS)), the pressure pain threshold (PPT), the grip strength and strength of the main arm and forearm muscle groups, and the functional outcome (measured by the Disability of Arm, Shoulder and Hand (DASH) and Patient-Rated Tennis Elbow Evaluation (PRTEE) questionnaires), were assessed before PRP injection and at one- and three-months follow-up. Flow cytometry measurements of the growth factors and inflammatory cytokines in PRP were performed, and the results were used to establish the relationship between those molecules and the clinical outcome. After three months from the intervention, the minimal clinically important difference in pain reduction and functional improvement was observed in 67% and 83% of patients, respectively. Positive correlations were found between the extent of pain reduction after three months and concentrations in the PRP of platelets, epidermal growth factor (EGF), vascular endothelial growth factor, and platelet-derived growth factors. The concentration of EGF in the PRP significantly correlated with an improvement in grip strength, strength of wrist extensors, and the size of functional improvement measured by the PRTEE. The local injection of PRP is a safe and effective treatment option for lateral elbow tendinopathy, and the clinical outcome is correlated with concentrations of its biologically active compounds.

Juvenile obesity in terms of various evaluation methods.

INTRODUCTION: Obesity is a civilization disease of the 21st century. The prevalence of obesity and overweight among children and adolescents is constantly increasing. BMI (body mass index) and WHR (waist to hip ratio) are methods of obesity assessment recommended by the WHO. Also, the WtHR (waist to height ratio), which takes into account height, is one of the most popular methods of diagnosing childhood obesity. A more recent diagnostic indicator is the FMI (fat mass index), which considers the percentage of the patient's body fat. THE AIM OF THE STUDY: was to compare the methods of assessing obesity in children and adolescents using the following indicators: BMI, WHR, WtHR, and FMI and to determine the consistency of the results obtained with them. MATERIAL AND METHODS: The study included 195 children aged 11-18 years, from whom the following data were collected: height, weight, waist circumference, and percentage of body fat. The calculated indices (BMI, WHR, WtHR, FMI), expressed in SDS, were compared using the Bland-Altman test, Passing-Bablok regression, and the slope chart. RESULTS: The fewest diagnoses of obesity were shown by FMI SDS (15.9%) and the highest by WHR SDS (28.7%). WHR SDS showed the least consistent results with BMI SDS. Significant statistical differences were found between BMI SDS and both FMI SDS and WtHR SDS. CONCLUSIONS: BMI, as the most acceptable obesity indicator, can be used as a screening method for assessing obesity. However, patients with boundary BMI values should be examined more precisely, using more than one index. FMI is recommended.

Familial hypercholesterolemia - treatment update in children, systematic review.

Familial hypercholesterolaemia is one of the most common genetic diseases, and its first symptoms occur in childhood. Proper diagnosis and treatment prevent young patients from severe consequences in their future. The treatment of this dyslipidaemia is still evolving, and new promising agents are being discovered. In this review we summarize the old and new treatment methods of familial hypercholesterolaemia, giving an update estimated on the latest publications.

Fractal Dimension and Texture Analysis in the Assessment of Experimental Laser-Induced Periodic Surface Structures (LIPSS) Dental Implant Surface-In Vitro Study Preliminary Report.

Laser-induced periodic surface structures (LIPSS) are the sub-wavelength periodic nanostructures generated by the femtosecond laser. Implant topography and its nanostructural changes can be important for biomedical applications. In order to compare the surface topography of different implants, appropriate mathematical and physical descriptive methods should be provided. The aim of the study was to evaluate the experimental LIPSS-based-Low Spatial Frequency LIPSS (LSFL) dental implant surfaces. Novel methods of surface analysis, such as Fractal Dimension Analysis and Texture Analysis, were compared to the standard surface roughness evaluation. Secondary, cell viability, and attachment tests were applied in order to evaluate the biological properties of the new titanium surface and to compare their correlation with the physical properties of the new surfaces. A Normal Human Dermal Fibroblast (NHDF) cytotoxicity test did not show an impact on the vitality of the cells. Our study has shown that the laser LIPSS implant surface modifications significantly improved the cell adhesion to the tested surfaces. We observed a strong correlation of adhesion and the growth of cells on the tested surface, with an increase in implant surface roughness with the best results for the moderately rough (2 µm) surfaces. Texture and fractal dimension analyses are promising methods to evaluate dental implants with complex geometry.

Celastrol and Resveratrol Modulate SIRT Genes Expression and Exert Anticancer Activity in Colon Cancer Cells and Cancer Stem-like Cells.

Metastatic colorectal cancer (CRC) remains a hard-to-cure neoplasm worldwide. Its curability declines with successive lines of treatment due to the development of various cancer resistance mechanisms and the presence of colorectal cancer stem cells (CSCs). Celastrol and resveratrol are very promising phytochemicals for colon cancer therapy, owing to their pleiotropic activity that enables them to interact with various biological targets. In the present study, the anticancer activities of both compounds were investigated in metastatic colon cancer cells (LoVo cells) and cancer stem-like cells (LoVo/DX). We showed that celastrol is a very potent anti-tumor compound against metastatic colon cancer, capable of attenuating CSC-like cells at the molecular and cellular levels. In contrast, resveratrol has a much greater effect on colon cancer cells that are expressing standard sensitivity to anticancer drugs, than on CSC-like cells. In addition, both polyphenols have different influences on the expression of SIRT genes, which seems to be at least partly related to their anti-tumor activity.

S16020 Pyridocarbazole Derivatives Display High Activity to Lung Cancer Cells.

BACKGROUND: Despite the dynamic development of medicine, globally cancer diseases remain the second leading cause of death. Therefore, there is a strong necessity to improve chemotherapy regimens and search for new anticancer agents. Pyridocarbazoles are compounds with confirmed antitumor properties based on multimodal mechanisms, i.e. DNA intercalation and topoisomerase II-DNA complex inhibition. One of them, S16020, displayed a wide spectrum of activity. OBJECTIVE: The aim of the study was to investigate the antitumor potency of six S16020 derivatives, synthesized according to the SAR (structure-activity relationship) method. METHODS: The biological evaluation included influence on cancer cell viability, proliferation, and migration, as well as P-glycoprotein activity. NHDF, A549, MCF-7, LoVo, and LoVo/DX cell lines were used in the study. RESULTS: All derivatives displayed low toxicity to normal (NHDF) cells at 1 and 2 µM (≤ 20% of cell growth inhibition). The highest reduction in cell viability was noted in A549 cells, which was accompanied by significant disruption of cells proliferation and motility. Compound 1 exhibited the strongest cytotoxic, antiproliferative, and antimigratory effects, higher than the reference olivacine. A significant reduction in P-glycoprotein activity was found for derivatives 6 and 1. CONCLUSION: S16020 derivatives could be considered as potential candidates for new anticancer drugs.

Unraveling the role of Breg cells in digestive tract cancer and infectious immunity.

Over the past two decades, regulatory B cells (Breg cells or Bregs) have emerged as an immunosuppressive subset of B lymphocytes playing a key role in inflammation, infection, allergy, transplantation, and cancer. However, the involvement of Bregs in various pathological conditions of the gastrointestinal tract is not fully understood and is the subject of much recent research. In this review, we aimed to summarize the current state of knowledge about the origin, phenotype, and suppressive mechanisms of Bregs. The relationship between the host gut microbiota and the function of Bregs in the context of the disturbance of mucosal immune homeostasis is also discussed. Moreover, we focused our attention on the role of Bregs in certain diseases and pathological conditions related to the digestive tract, especially Helicobacter pylori infection, parasitic diseases (leishmaniasis and schistosomiasis), and gastrointestinal neoplasms. Increasing evidence points to a relationship between the presence and number of Bregs and the severity and progression of these pathologies. As the number of cases is increasing year by year, also among young people, it is extremely important to understand the role of these cells in the digestive tract.

Occult Autoimmune Background for Epilepsy-The Preliminary Study on Antibodies Against Neuronal Surface Antigens.

Objective: The objective of the study was to determine the incidence of antibodies against neuronal surface antigens (NSA-ab) in patients with different types of epilepsy, in comparison with the subjects diagnosed with immune-mediated disorders. Methods: Forty patients with drug-resistant epilepsy (DRE) of unknown origin, 16 with post-stroke epilepsy, and 23 with systemic autoimmune disorders (SAD) with CNS involvement were included. NSA-ab were sought in serum using indirect immunofluorescence method. Relationships were analyzed between presence of NSA-ab and clinical presentation. Results: NSA-ab was detected in the sera from five patients: anti-DPPX in one patient, anti-AMPAR1/R2 in two, anti-LGI1 in one and, in one case, both anti-CASPR2 and DPPX IgG. Out of these five patients, three represented the SAD subgroup and two the DRE subgroup. None of the patients with post-stroke epilepsy was positive for NSA-ab. Significance: Autoimmune etiology is worth considering in patients with drug-resistant epilepsy of unknown origin. The presence of NSA-ab in patients with systemic autoimmune disorders may be caused by unspecifically enhanced autoimmune reactivity. NSA-ab seem not to be related to epilepsy resulting from ischemic brain injury.

Three dimensional in vitro culture systems in anticancer drug discovery targeted on cancer stem cells.

Worldwide, tumors are one of the most common causes of death. Every year 3.7 million new cases occur in Europe and more than 1.9 million patients die (WHO data). Most of the fields of research are focused on developing new therapeutic strategies that will be effective in eliminating the tumor, preventing its remission, and avoiding or reducing the side effects of therapy. In the past, generally classical 2D cell cultures or immunodeficient animal models had been used to cultivate and test drugs on human cancer cell lines. Nowadays, there are increasing interests in three-dimensional (3D) cell cultures, a method with significant differences from flat cultured cells, both considering gene expressions and cell-cell interactions. Various evidence suggests that high tumorigenic properties might be dependent on the occurrence of a small cell population, pointed out to be responsible for metastasis and recurrence. This population is called cancer stem cells (CSCs), hinted to have a lot of similarities with normal stem cells. CSCs are the main reason for chemotherapy failure as well as multi-drug resistance (MDR). CSCs can also interact through the cytokine network, with other cells like the macrophages of the inflammatory system. The big advantage of a 3D culture is the possibility to isolate and investigate the CSCs population surrounded by its environment. This article aims to sum up known 3D cell cultures, especially in the field of CSCs research due to the importance of the tumor's environment on stem cell's markers expression and their development.