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Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease which is characterized by death of motor neurons, development of muscle weakness, paralyses and atrophy of skeletal muscles. The most common immediate cause of lethal outcome of ALS patients is respiratory insufficiency due to paralysis of respiratory muscles. Effectiveness of currently used medications for ALS treatment is very limited and can extend life of patient only for few months. One of perspective directions in development of ALS therapy is use of microvesicles prepared from mesenchymal stem cells (MSC). Such microvesicles have neuroprotective properties and some advantages in comparison with parental cells (lower immune rejection, no tumor risk, etc.). Earlier we found that transplantation of MSC-derived microvesicles increases survival and reduces the severity of motor disorders in mice with ALS model. In current paper we evaluated the influence of transplantation of microvesicles obtained from mesenchymal stem cells overexpressing vascular endothelial growth factor (VEGF) and angiogenin (ANG) on contractile characteristics of diaphragmatic muscle of FUS transgenic mice with ALS model. Electrically induced single (0,1 Hz stimulation) and tetanic (5-50 Hz) muscle contractions were recorded with use of standard myographic technique on diaphragmatic muscles of 3,5-month old FUS mice after single or two time transplantation of microvesicles derived from MSC overexpressing VEGF and ANG. It was found that relative forces of single and tetanic muscle contractions were significantly increased in FUS mice received single time transplantation of microvesicles carrying VEGF and ANG comparing to FUS mice without treatment. No significant effects of treatment with microvesicles carrying VEGF and ANG on the threshold of diaphragmatic muscle contraction was found. Thus, transplantation of microvesicles carrying VEGF and ANG increases the contractility of diaphragmatic muscle of FUS mice, which should be beneficial in ALS-modelled pathology and can be one of mechanisms of therapeutic effects of microvesicles carrying VEGF and ANG on FUS mice with ALS model.

Rapid analysis of milk components is often necessary in clinical settings, biotechnology, the food industry, and animal husbandry. Existing data on the possibility of analyzing milk through a correlation between lactose and mineral concentrations remain contradictory and require further verification. This study examines a possible correlation between the electromotive force (EMF) readings of ion-selective electrodes and lactose in raw cow's milk. A laboratory setup with an ion meter-conductometer-thermometer equipped with a potassium-selective electrode, a heated magnetic stirrer, and ultrasonic and viscometric milk quality analyzers was used to collect measurements of various milk parameters. Milk with the following parameters was used: fat content (from 2.59% to 4.49%), protein content (from 3.01% to 3.72%), and lactose (from 4.11% to 5.11%). As a result, it was found that changing the temperature from 30 °C to 39 °C leads to a 3-5% decrease in the generated EMF of a milk sample. The EMF decrease occurs differently for milk with different combinations of quality parameters. A statistically significant strong linear relationship of r = 0.88 at p < 0.05 was established, as well as a determination coefficient of R2 = 0.77 between lactose content and EMF in milk. This result opens the possibility of rapid analysis of milk using potentiometric methods.

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the accumulation of alpha-synuclein. Growing evidence implicates the gut-brain axis in PD pathogenesis, with dysbiosis potentially influencing alpha-synuclein homeostasis and serving as a disease biomarker. Methods: We established substrate-specific bacterial enrichment cultures from a pooled fecal inoculum of 15 treatment-naïve PD patients. 16S rRNA gene sequencing defined the taxonomic profiles of five consortia (PLactate, MTryptone, MPeptone/Sucrose, MFat, MStarch). The impact of bacterial lysates on a human neuronal model was assessed using SH-SY5Y neuroblastoma cells. Gene expression of SNCA, HSPA8, SNAP25, STX1A, and APP was quantified via qRT-PCR, and cytotoxicity was measured by MTT assay. Results: The choice of carbon source selected for communities with distinct taxonomic profiles. Lysates from the MTryptone consortium, dominated by Clostridium, Coprococcus, and Eggerthella, significantly upregulated APP, SNAP25, and HSPA8 expression and increased cytotoxicity in live co-culture. Conversely, lysates from the MPeptone/Sucrose consortium, enriched in Lactobacillus, Bifidobacterium, Peptoniphilus, Ruminococcus, and Bacteroides, downregulated SNAP25, HSPA8, and STX1A and exhibited a protective effect on viability. SNCA expression remained unchanged across all treatments. Conclusions: Our findings demonstrate that specific gut bacterial consortia derived from PD patients can directly and differentially modulate the expression of key neuronal genes linked to synaptic function and protein processing. This work provides direct in vitro evidence supporting the hypothesis that a shift in gut microbial ecology may contribute to neuronal dysfunction in PD, highlighting specific microbial guilds as potential targets for therapeutic intervention.

Nanomaterial-based drugs are currently being actively developed for tumor diagnostics and therapy, and some are already in clinical use. However, systemic administration of nanomedicines often results in their rapid removal from the bloodstream by cells of the mononuclear phagocytic system and accumulation in the liver and spleen. For abdominal and pelvic tumors, intraperitoneal administration may be an option, overcoming the inefficiency of passive delivery. Peritoneal immune cells, which are tropic to tumor nodes, can be used to transport nanoagents to tumor sites. In this study, biocompatible complexes based on upconversion nanoparticles (UCNP) were synthesized and their interaction with mouse peritoneal macrophages was studied. UCNP-based particles were shown to be capable of being taken up by macrophages and maintain their integrity for an extended period. The obtained results allow us to further consider the use of peritoneal macrophages as a system for delivering nanopreparations to tumor foci.

Various polysaccharide-based compositions containing xanthan and alginate were developed to produce spherical gel particles with immobilized probiotic microorganisms, Lactobacillus plantarum, and the bacteriocins produced by this strain. The conditions for the formation of spherical gel particles were optimized, and their stability was evaluated under different pH values and temperature conditions. It was demonstrated that at pH 2.4 the gel particles did not undergo degradation; thus, under conditions simulating gastric juice, lactic acid bacteria retained their viability. At pH values close to 7.5–8.0, visual observation showed dissolution of the gel particles, indicating that under conditions mimicking the colon environment, immobilized microorganisms were released from the matrix. In addition, L. plantarum actively synthesized bacteriocins exhibiting inhibitory activity against test pathogenic microorganisms. Thus, the developed gel particles containing probiotic cultures can effectively deliver viable probiotics to the colon without degradation in the stomach.

This study compared the effects of citric acid (CA) derived from microbial fermentation using Aspergillus niger (400 mg kg-1) and natural extraction from Citrus limon (400 mg kg1) in albino mice. Parameters assessed included cytogenetic damage via the micronucleus (MN) assay, immunological response through immunoglobulin (IgA, IgG, IgM) titers, and hepatoprotective activity based on liver enzyme levels (AST, ALT, ALP) and histopathological investigation. Mice were divided into six groups (n = 10/group) as follows: (1) negative control, (2) carbon tetrachloride (CCl₄ 0.02%), (3) lemon CA, (4) A. niger, (5) CCl₄ + lemon CA, and (6) CCl₄ + A. niger CA. The CCl4 group exhibited a significantly higher MN frequency (0.06 ± 0.008) compared to the control (0.01 ± 0.001). Co-treatment with lemon CA and A. niger CA significantly reduced MN frequencies to 0.02 ± 0.005 and 0.03 ± 0.001, respectively. Immunologically, lemon CA significantly increased IgG and IgM titers but decreased IgA compared to the control. Both CA sources demonstrated hepatoprotective effects, significantly attenuating the CCl4-induced elevation of AST, ALT, and ALP and reducing histopathological damage. The results indicate that while both CAs offer protective benefits, lemon-derived CA was more effective at the tested doses, particularly in mitigating genotoxicity and modulating the immune response. 

Phenols are used to make detergents, some prescription drugs, pesticides, and other industrially manufactured products. Consuming a range of phenolic compounds found in food reduces the risk of health issues because they act as antioxidants. Paclitaxel is a commonly used treatment for a variety of malignancies. It has been shown that higher doses of paclitaxel are more successful in treating a variety of cancer types. Nevertheless, in addition to inhibiting the growth of cancerous cells, chemotherapy medications have some unfavorable side effects. Twenty-four albino male rats were divided into three groups, and each group consisted of eight rats. Group I: negative control, injected with normal saline. Group II: positive control, received paclitaxel for 4 doses at 7 mg/kg over 30 days without phenol treatment. Group III: rats were treated with crude phenol extract at 10 mg/kg administered orally, along with paclitaxel, and samples were collected after 30 days. The results showed that the crude phenol extract reduced creatinine, urea, uric acid, AST, ALT, and ALP activity, while it increased SOD levels. The rats treated with crude phenol extract showed improvement in liver and kidney histopathological alterations. In conclusion, we investigated the possible advantages of crude phenol extract from Cinnamomum cassia L. in reducing the side effects of paclitaxel, a chemotherapy drug.

The development of photopolymer 3D printing materials for biomedical applications requires the creation of a new class of polymer resins, that combine high strength, antibacterial activity, and low toxicity against eukaryotic cells. In this paper, a new material for 3D printing based on polymethyl methacrylate (PMMA) and tellurium nanoparticles (NPs) was developed. NPs Te were synthesized by laser ablation in water, transferred in acetone, and then included in a MMA solution. NPs concentrations Te were 0.001, 0.01, and 0.1% by volume. The introduction of Te NPs did not alter the energy required for polymerization or the mechanical properties of the finished polymer; however, it increased the degree of final polymerization as indicated by FTIR data. Additionally, it added significant bacteriostatic properties without increasing toxicity against eukaryotic cells. The mechanism of antibacterial action can be mediated through the induction of oxidative stress in bacteria (increased generation of 8-oxoGua and long-lived reactive forms of proteins in aqueous solutions). The severity of the antibacterial action is determined by the dose of the introduced NPs Te.

Acute limb ischemia (ALI) is characterized by a sudden reduction in arterial blood flow and may result in gangrene, systemic complications, and death. Urgent revascularization can be limb- and life-saving; however, it is associated with a risk of early postoperative complications. Both perioperative and intraoperative risk factors contribute to these outcomes and are influenced by the selected method of revascularization. The aim of this study was to assess the impact of intraoperative risk factors on postoperative outcomes according to the chosen treatment modality for acute limb ischemia, namely open or endovascular revascularization. Patients with Rutherford class II ALI from a single population were divided into two groups comparable in clinical and demographic characteristics: Group I – open surgical treatment (n = 50) and Group II – endovascular revascularization (n = 50). The influence of identified intraoperative risk factors on the development of complications differed between the two revascularization approaches. An operative time exceeding 85 minutes in the open surgery group (2.73) and exceeding 92 minutes in the endovascular group (1.52) was associated with an increased risk of adverse postoperative outcomes. The urgent nature of the procedure was also an independent predictor of adverse events in both groups, with ORs of 12.11 and 9.13 for Groups I and II, respectively. Outcomes of ALI treatment and the early postoperative course in the overall population depend on the selection of the revascularization method in accordance with each patient’s individual perioperative risk profile. Comparison of endovascular and open approaches demonstrated differences in outcomes favoring endovascular revascularization only in a specific subgroup of patients with a less severe functional class of limb ischemia.

Copy number variations (CNVs) are involved in eukaryotic genomes evolution. CNVs also the causes of multifactorial and monogenic diseases. Retroelements can be causes of CNVs emergence and evolution. Retroelements occupy 42.5% of the human genome and are located mainly in intergenic, intronic and regulatory regions. The mechanisms of CNVs formation are due to non-allelic homologous recombination between retroelements due to their distribution across all chromosomes and the presence of homologous sequences. Role of recombinations between retroelements as causes of neurofibromatosis type 1, Cowden syndrome, Peutz-Jeghers syndrome, Langer-Gideon syndrome, Fanconi anemia, amelogenesis imperfecta, and spastic paraplegia type 4 and other monogenic diseases is described. Mechanisms of CNVs formation using retroelements suggest the role for retroelements as causes of trinucleotide repeat expansion diseases. The influence of LINE1 in Huntington's chorea caused by CAG expansion is described. In fragile X syndrome, CGG expansion occurs in the FMR1 gene, which homologue in Drosophila is involved in retroelements inhibition. In humans, CGG-binding protein influence on Alu and LINE1 described. Role for Alu in GAA expansion identified in Friedreich's ataxia. These relationships suggest the possible retroelements role in triplet code development during the origin of life. At the human population level, the underlying mechanisms of evolution through the influence of retroelements are the cause of numerous diseases. However, in the wild, the same mechanisms serve as substrates for the natural selection of more adaptive traits and eukaryotes evolution.

Gamma-wave stimulation represents one of the most promising non-pharmacological approaches for enhancing cognitive function, which declines with aging and in neurodegenerative diseases. The aim of the present study was to identify optimal protocols for maximizing the effectiveness of gamma-wave stimulation. We investigated the effects of various combinations of physical and cognitive loads during visual gamma-wave stimulation at a frequency of 40 Hz using light of different wavelengths on the amplitude of gamma rhythms in the human brain. The effects were evaluated both at the level of individual EEG leads and across global brain electrical activity. The strongest response to visual gamma stimulation was observed in the occipital region. This effect exhibited a cumulative component, reflected by an increase in baseline activity at the stimulation frequency. The cumulative effect was most pronounced in the occipital and temporal regions. The magnitude of the gamma-stimulation response depended on the wavelength of the light stimulus and increased in the order of white, pink, and red. The most robust effect was observed with red-light stimulation combined with physical exercise. The influence of cognitive load was more prominent in the frontal and parietal regions of the brain.

Polycystic ovary syndrome (PCOS) is a multifactorial endocrine disorder with significant genetic components. Vitamin D binding protein (VDBP) regulates vitamin D bioavailability, and its gene polymorphisms may influence PCOS susceptibility. This study investigated the correlation of VDBP SNPs rs4588 and rs7041 with the risk of developing PCOS in Iraqi women. A case-control study involving 60 PCOS patients and 30 healthy controls was conducted. Serum vitamin D and VDBP levels were measured, and VDBP SNPs (rs4588, rs7041) were genotyped by polymerase chain reaction (PCR). The results showed that the levels of VDBP were significantly lower in individuals with PCOS compared to healthy individuals, with a statistically significant difference (p < 0.05). The frequencies of A and G alleles were 0.46 and 0.54 in PCOS while 0.47 and 0.53 in the control, respectively. The homozygous GG genotype and the heterozygous AG genotype were not statistically significant in PCOS patients compared to the controls. Furthermore, the frequencies of the A and C alleles for the rs7041 SNP were 0.47 and 0.53 in PCOS, respectively, while they were 0.42 and 0.58 in the control group. When compared to the control group, PCOS patients' homozygous AA and heterozygous AC genotypes showed statistically significant differences (p < 0.05). Conversely, there are no appreciable variations in the prevalence of homozygous CC genotypes between PCOS patients and controls. The findings of the study revealed that the rs7041 polymorphisms in the VDBP gene may contribute to increasing the risk of PCOS.

Resveratrol is a phytoalexin naturally produced by several plants in response to injury or pathogenic attack, such as bacterial or fungal infections. In this study, approximately 0.5 kg of fresh black grape skin cultivated in Iraq was macerated in 80% ethanol for three days. Hydrolysis was performed using 10% HCl at 60 °C, followed by filtration. The filtrate was then extracted with chloroform. The organic layer was evaporated to dryness, and the residue was subjected to further purification using silica gel G60 in an open column with a mobile phase consisting of benzene:methanol:acetic acid (20:4:1). Final purification was carried out using preparative layer chromatography (PLC). The resulting pure resveratrol (Res) was dried and stored in a cool, dark place for further analysis using UV absorption, TLC, HPLC, and FTIR spectroscopy. Five groups of albino mice (n = 5 per group) were used in the study. Four groups were rendered hyperglycemic by administration of alloxan. The animals were then treated intraperitoneally for two weeks as follows: Group 2 received Res at a dose of 5 mg/day; Group 3 received Res at 0.25 mg/day; Group 4 received glibenclamide at 600 µg/kg/day; Group 1 consisted of normal non-diabetic animals, and Group 5 represented untreated diabetic controls. The effects of resveratrol on liver function were evaluated by measuring blood glucose levels, serum malondialdehyde (MDA), and superoxide dismutase (SOD) as indicators of antioxidant activity. Additionally, in vitro kinetic assays were conducted to determine serum levels of aspartate aminotransferase (AST/SGOT) and alanine aminotransferase (ALT/SGPT).

The hydroperoxyl radical (•OOH), a reactive oxygen species (ROS), plays a critical role in regulating physiological processes. Its involvement in cellular redox pathways and association with pathological processes make it a key molecule for understanding how excessive free radicals modulate cellular metabolism. This study investigated the effects of exogenous •OOH on the redox metabolism of mononuclear cells during phagocytosis. Rat-derived mononuclear cells were treated with different concentrations of •OOH (generated via spark-discharge non-coherent radiation), and phagocytosis was induced using latex particles. The effect of •OOH on cell membrane integrity was assessed by trypan-blue-staining, while redox metabolites were quantified by spectrofluorimetry and spectrophotometry at 1-, 30-, and 60-minutes post-phagocytosis induction. In unexposed cells, free FAD and NADH levels increased during phagocytosis, tryptophan-containing proteins remained stable, and glycation end-products (AGEs) rose significantly by 60 minutes post-phagocytosis induction. •OOH at concentrations of (3.6 ± 0.9) × 10⁻⁵ and (1.8 ± 0.45) × 10⁻⁴ mol/L did not alter redox metabolism. However, exposure to higher concentrations ((5.4 ± 1.35) × 10⁻⁴ mol/L) induced significant cytotoxic effects. At the metabolic level, this dose triggered a marked redox imbalance, evidenced by a 1.4-fold increase in FAD, altered coenzyme ratios (NADH/FAD and NADH/NAD), and a significant decrease in tryptophan-related protein fluorescence, indicating extensive macromolecular damage. All •OOH concentrations tested suppressed the formation of AGEs 60 minutes after phagocytosis induction. We propose that this inhibition is not due to a protective effect, but to fragmentation of the protein backbone or modifications in the side chains induced by radicals.

Noroviruses are the leading cause of outbreaks of nonbacterial gastroenteritis and the second most common cause of all viral intestinal infections. An effective norovirus vaccine is expected to help reduce the incidence of intestinal infections, but intensive efforts to develop such a vaccine have so far been unsuccessful. Failures in vaccine development may be due to the high heterogeneity of noroviruses and/or the hypothetical low protective efficacy of the immune response to the most common virus variants, such as GII.4 Sydney 2012. The subject of the study was potential vaccine components – virus-like particles (VLPs), formed from VP1 of norovirus GII.4 Sydney 2012 (VP1N) and VLPs from a fragment of this protein containing the shell domain and hinge region (SN). We investigated the effect of VLPs on the ability of human dendritic cells (DCs) to recruit T cells into the immune response in vitro. VLP-treated DCs were cultured with pure CD4⁺ T cells, and then T cell maturity and cytokine production were assessed. It has been shown that VP1N-treated DCs, but not SN-treated DCs, have an increased ability to shift the ratio of T cell from naïve T cells to more mature central memory T cells and stimulate IL-17 production. Intracellular cytokine assay revealed no differences in T-helper type 1 (Th1), Th17 and Th1/17 levels between mixed cultures with VP1N-treated DCs and control DCs. Apparently, the increase in IL-17 production occurs due to an increase in the activity of mature Th17, and not the maturation of new producer cells.

Cardiovascular diseases (CVDs) are the leading cause of mortality worldwide. Heart failure (HF), a major pathology of the myocardium, is characterized by impaired cardiac function that leads to an abnormal enlargement of the heart, known as hypertrophy. In the study of the molecular mechanisms of HF pathogenesis, animal models play a crucial role. To characterize induced HF in animal models, biochemical approaches, such as quantifying the concentration of biomarkers in blood serum, are extremely important. Here we report a new immunochemical test system based on the measurement of the concentration of the B-type natriuretic peptide, protein biomarker of HF and hypertrophy, that can be utilized for characterization of HF development in rats and serve as a tool for further BNP concentration analysis.

Alzheimer’s disease (AD) is a multifactorial neurodegenerative disorder in which neuroinflammation plays a major role in its pathogenesis, alongside the formation of amyloid plaques and neurofibrillary tangles. Necroptosis, a recently discovered regulated form of cell death mediated by the kinases RIPK1 and RIPK3, is considered one of the mechanisms contributing to neuroinflammation and neuronal death in AD. In this study, we evaluated the effect of chronic necroptosis inhibition using Necrostatin-1, a RIPK1 blocker, on the progression of neurodegeneration in aged 5xFAD mice, a model of the familial form of AD. Over a 12-week treatment period, the animals’ neurological status was assessed, followed by evaluation of long-term memory using the Morris water maze test, histological analysis of the prefrontal cortex and hippocampus, and RT-PCR analysis of the expression of key genes associated with necroptosis and inflammation. Chronic administration of Nec-1 significantly slowed the progression of neurological deficits in both male and female 5xFAD mice. Inhibition of necroptosis prevented the loss of normal neurons, reduced the number of hyperchromic cells, and decreased the severity of pericellular and perivascular edema in the examined brain regions. However, in the Morris water maze test, learning and memory improved only partially in males, but not in female 5xFAD mice. This may be attributed to the increased expression of the anti-inflammatory cytokine IL-10 in the cortex and hippocampus of males. The results obtained indicate that inhibition of necroptosis by Necrostatin-1 represents a promising therapeutic approach for correcting neurological impairments and mitigating morphological brain alterations in Alzheimer’s disease.

Primary lactose non-persistence (LNP, hypolactasia) represents an autosomal-recessive condition, which is mainly attributed to the presence of the ancestral C-allele in the MCM6 -13910C>T (rs4988235) variant regulating the expression of the lactase (LCT) gene in individuals of European ancestry. Since the studies linking lactose intolerance C-allele and caloric accumulation, as well as gastrointestinal symptoms remain ambiguous to date, we decided to examine for a possible relation between the rs4988235 lactose intolerance allele and certain health parameters (body mass index, waist circumference, breastfeeding duration, and existing gastrointestinal tract diseases) in individuals (N = 2912) from four regions of the Volga-Ural region (VUR), i.e., Sverdlovsk Oblast, Republic of Bashkortostan, Chelaybinsk Oblast, and Udmurt Republic. In addition, we sought to clarify the genotype and allele frequencies of the MCM6 rs4988235 in the large sample from the VUR based on territorial and ethnic specificity. While examining a relation between several health conditions and the MCM6 variant, we determined a link between the rs4988235 CC genotype and prolonged breastfeeding duration (more than 1 year) in the total sample (OR = 1.63, 95%CI 2.32–5.88, p = 0.005) and in Russians (OR = 1.75, 95%CI 2.85–4.16, p = 0.024), which became more significant in individuals with a full-term period of gestation (more than 36 weeks) (OR = 1.67, 95%CI 1.13–2.45, p = 0.009) and was positively associated with higher birth weight (β = 3.11, p = 1.8x10^-3). Findings obtained point to a compensatory effect of prolonged breastfeeding on diminishing manifestation of genetically predisposed primary hypolactasia.

Bacteria of the species Streptococcus pneumoniae, dominant in the etiological structure of community-acquired pneumonia in children, are represent a serious problem in the field of human infectious pathology. Detailed molecular and genetic characteristics of eleven S. pneumoniae strains isolated from sputum samples of children with community-acquired pneumonia were obtained using whole genome sequencing and bioinformatic analysis. Based on the analysis of the nucleotide sequences of seven housekeeping genes (aroE, gdh, gki, recP, spi, xpt, ddl), it was found that S. pneumoniae strains included in the study belong to six sequence types: ST1367, ST819, ST1262, ST180, ST15069, ST66. Using SeroBA algorithm S. pneumoniae strains were assigned to five serotypes:11C, 22F, 15C, 9N, 3. Genome annotation using ResFinder and CARD databases allowed us to identify determinants of resistance to macrolides (ermB, RImA), fluoroquinolones (patA, patB, pmrA), lincosamides (RImA), aminoglycosides (aph(3')-Ia) and tetracyclines (tetM and tet32). A high frequency of detection of pathogenicity genes encoding choline-binding proteins, fibronectin-binding proteins, pneumolysin, autolysin, hyaluronidase, neuraminidase, capsule proteins, zinc metalloproteinase in the genome of S. pneumoniae strains was noted.  Phylogenetic analysis of the nucleotide sequences of the genome of Nishny Novgorod strains and strains deposited in GenBank/NCBI showed a high level of genetic variability of pneumococci circulating both in Russia and abroad.

Vitex negundo is a medicinal plant renowned for its wide spectrum of therapeutic properties, including significant anticancer activity. This study explores the inhibitory effects of V. negundo methanolic extract and essential oil on α-amylase purified from the serum of Iraqi lung cancer patients, as well as their cytotoxic effects on the A549 human lung cancer cell line. α-Amylase, an enzyme involved in carbohydrate metabolism, exhibits altered activity under cancerous conditions, making it a potential therapeutic target. The enzyme was purified through a multi-step procedure comprising ammonium sulfate precipitation, dialysis, ion-exchange chromatography, and gel filtration chromatography. This process resulted in an increase in specific activity from 0.94 to 17.0 U/mg protein. Both extracts of V. negundo were assessed for their capacity to inhibit α-amylase activity and reduce the viability of lung cancer cells. The methanolic extract demonstrated a more pronounced inhibitory effect on α-amylase (88.4% at 10 µg/mL) compared to the essential oil (77.0% at 10 µg/mL). Moreover, MTT assay results indicated concentration -dependent cytotoxicity against A549 cells, with IC₅₀ values of 68.79 µg/mL for the methanolic extract and 81.14 µg/mL for the essential oil. These findings underscore the potential of V. negundo—particularly its methanolic extract—as a promising natural anticancer agent, capable of targeting cancer-related metabolic pathways and suppressing tumor cell viability. The results warrant further investigation into the development of V. negundo-based therapeutics for lung cancer treatment.

Chronic lymphocytic leukaemia (CLL) is distinguished from other lymphoid tumours due to the fact that its biology and clinical symptoms are very different from those of other lymphoid cancers. In the past ten years, there has been significant progress made in the understanding of the pathogenesis of the disease. In the field of cancer research, there have been numerous significant areas that have been investigated. The mechanisms of genetic vulnerability, the role of immunogenetic factors in the development of disease, genomic changes, epigenetic subtypes, epigenomic reprogramming of tumour cells, the control of interactions between tumour cells and their environment, and the dynamics of clonal evolution from monoclonal B cell lymphocytosis to diffuse large B-cell lymphoma are some of the topics that are investigated. As a result of the accumulation of information, new targeted drugs and management strategies have been developed, which has resulted in the opening of new therapy avenues. The purpose of this review is to examine the patterns of DNA methylation, immunological markers, and molecular and immunological characteristics that are present in patients who have chronic lymphocytic leukaemia.