Type Enzyme Research Articles

Liquiritigenin inhibits the migration, invasion, and EMT of prostate cancer through activating ER stress

Liquiritigenin (LQ) is a monomeric compound found in licorice, a leguminous plant, and has been reported to exhibit antitumor effects in various lines of cancer cells. However, the underlying molecular mechanisms by which LQ exerts its antitumor effects remain largely unknown. In this study, the effects of LQ on the migration, invasion, and epithelial-mesenchymal transition (EMT) of prostate cancer (PCa) cells were investigated. We found that LQ effectively inhibited the migration and invasion of PCa cells in vitro, and this effect was further confirmed in xenograft lung metastasis models. In addition, LQ was found to activate endoplasmic reticulum stress (ER stress) in PCa cells. Further studies found that LQ upregulated the expression of inositol-requiring enzyme type 1α (IRE1). When IRE1 was knocked down, we observed a weakened inhibitory effect of LQ treatment on the migration and invasion of PCa cells. This observation suggests that LQ may inhibit the migration, invasion and EMT of PCa cells through activating the IRE1 branch of ER stress. In conclusion, our research may provide a novel therapeutic strategy for PCa.

The Identification and Characterization of the Resistance Genes of Klebsiella pneumoniae Plasmids Transformed in E. coli BL21(DE3)

The emergence of multidrug-resistant Klebsiella pneumoniae constitutes a serious threat to global public health due to the limited treatment options available. This study aimed to identify some characteristics carried on the plasmids of K. pneumoniae isolated from pathological cases. The plasmid was extracted from K. pneumoniae and transformed into E. coli BL21(DE3) by the heat shock method using 0.1 M CaCl2. The results showed that transformed E. coli BL21(DE3) became resistant to four types of antibiotics which included cefepime (FEP) 10 μg, ceftazidime (CAZ) 30 μg, tetracycline (TE) 10 μg and rifampin (RA) 5 μg. In addition, transforming colonies of E. coli BL21(DE3) were able to producer biofilm as well as production of urease enzyme. The extracted plasmids were digested by 3 types of restriction enzymes (enzyme EcoRI, BamHI and Hind III) to know the restriction sites available in bacterial plasmids. The results showed the presence of some EcoR1 restriction sites by observing the change in the physical shape of the plasmid after being treated with the EcoR1. Meanwhile, no restriction sites were observed for BamH1 and HindIII.

Gaining Insight into the Catalytic Mechanism of the R132H IDH1 Mutant: A Synergistic DFT Cluster and Experimental Investigation.

Human isocitrate dehydrogenase 1 (IDH1) is an enzyme that is found in humans that plays a critical role in aerobic metabolism. As a part of the citric acid cycle, IDH1 becomes responsible for catalyzing the oxidative decarboxylation of isocitrate to form α-ketoglutarate (αKG), with nicotinamide adenine dinucleotide phosphate (NADP+) as a cofactor. Strikingly, mutations of the IDH1 enzyme have been discovered in several cancers including glioblastoma multiforme (GBM), a highly aggressive form of brain cancer. It has been experimentally determined that single-residue IDH1 mutations occur at a very high frequency in GBM. Specifically, the IDH1 R132H mutation is known to produce (D)2-hydroxyglutarate (2HG), a recognized oncometabolite. Using the previously determined catalytic mechanism of IDH1, a DFT QM model was developed to study the mechanistic properties of IDH1 R132H compared to wild type enzyme. Validating these insights, biochemical in vitro assays of metabolites produced by mutant vs wild type enzymes were measured and compared. From the results discussed herein, we discuss the mechanistic impact of mutations in IDH1 on its ability to catalyze the formation of αKG and 2HG.

Exploration of SOD3 from gene to therapeutic prospects: a brief review.

Superoxide dismutase 3 (SOD3) is a type of antioxidant enzyme, which plays an important role in converting superoxide anion into hydrogen peroxide through its extracellular activity. This enzyme has been widely studied and evaluated from various points of view, including maintaining cellular redox balance, protecting against oxidative damage, and enhancing overall cellular resilience. The current paper focuses on SOD3 expression from a functional perspective. In addition to a detailed examination of the gene and protein structure, we found ample evidence indicating that the expression level of SOD3 undergoes alterations in response to various transcription factors, signaling pathways, and diverse conditions. These fluctuations, by disrupting the homeostasis of SOD3, can serve as crucial indicators of the onset or exacerbation of specific diseases. In this regard, significant efforts have been dedicated in recent years to the treatment of diseases through the regulation of SOD3 expression. The ultimate goal of this review is to extensively highlight and demonstrate the immense potential of SOD3 as a therapeutic target, emphasizing its profound impact on health outcomes.

Sequence analysis, homology modeling, tissue expression, and potential functions of seven putative acetylcholinesterases in the spider Cupiennius salei.

Acetylcholine esterases (AChEs) are essential enzymes in cholinergic synapses, terminating neurotransmission by hydrolysing acetylcholine. While membrane bound AChEs at synaptic clefts efficiently perform this task, soluble AChEs are less stable and effective, but function over broader areas. In vertebrates, a single gene produces alternatively spliced forms of AChE, whereas invertebrates often have multiple genes, producing both enzyme types. Despite their significance as pesticide targets, the physiological roles of invertebrate AChEs remain unclear. Here, we characterized seven putative AChEs in the wandering spider, Cupiennius salei, a model species for neurophysiological studies. Sequence analyses and homology modeling predicted CsAChE7 as the sole stable, membrane-bound enzyme functioning at synaptic clefts, while the others are likely soluble enzymes. In situ hybridization of sections from the spider's nervous system revealed CsAChE7 transcripts co-localizing with choline acetyltransferase in cells that also exhibited AChE activity. CsAChE7 transcripts were also found in rapidly adapting mechanosensory neurons, suggesting a role in precise and transient activation of postsynaptic cells, contrasting with slowly adapting, also cholinergic, neurons expressing only soluble AChEs, which allow prolonged activation of postsynaptic cells. These findings suggest that cholinergic transmission is influenced not only by postsynaptic receptors but also by the enzymatic properties regulating acetylcholine clearance. We also show that acetylcholine is a crucial neurotransmitter in the spider's visual system and sensory and motor pathways, but absent in excitatory motor neurons at neuromuscular junctions, consistent with other arthropods. Our findings on sequence structures may have implications for the development of neurological drugs and pesticides.

Saccharification of Different Delignified Sawdust Masses from Various Trees Along the Lagos Lagoon in Nigeria

Sawdust, a major waste product of the forestry industry, is accumulating along the Lagos Lagoon in Lagos, Nigeria, without it being effectively managed. Besides its use in The saccharification of sawdust could contribute to the development of renewable energy sources and feedstock for bioproduct development. The process is, however, not that straightforward as variables such as the type of cellulase enzyme, pretreatment of the cellulose substrate, and optimizing of cellulase to cellulose ratio are a few that need to be optimized for the process to be effective in terms of glucose production.manufacturing sound-absorbing boards to reinforce concrete beams and for energy purposes, its potential as a renewable energy source and feedstock for bio-product development has not yet been realized. Cellulose, a glucose biopolymer and structural component of cellulose can be hydrolyzed by a hydrolytic enzyme known as cellulase. During the process, the enzyme breaks the B-1,4-glucosidic bond, which keeps the glucose units together, and by acting on this bond, numerous glucose units are released. As part of sawdust, the cellulose molecule is not freely available for the degradation action of the cellulase enzyme as it is strongly associated with lignin, which acts as bio-glue, keeping cellulose and hemicellulose together. Delignification is an effective technique that was used to make the sawdust from ten different trees along the Lagos Lagoon in Nigeria more susceptible to saccharification by cellulase isolated from the fungus Aspergillus niger. Delignified and non-delignified sawdust masses between 2 mg and 10 mg were incubated with the A. niger cellulase solution (2 mg.mL-1), whereafter, the amount of sugar produced by the cellulase action was determined. The percentage saccharification of each sawdust material was also linked with the amount of sugar produced during cellulase action. From these investigations was concluded that delignification increased sugar production when almost all the masses of different sawdust materials were degraded. It was also observed that the ratio of sawdust mass to enzyme concentration is an important variable that influences the effectiveness of the saccharification process. The percentage saccharification of the various sawdust materials was also determined, and it indicated that the highest percentage of saccharification was not obtained when the highest amount of sawdust was degraded, producing the highest amount of sugar.

Innovation in Bromelain Enzyme Production from Pineapple and Honey and Its Characteristics

Bromelain enzyme is one type of protease enzyme that has the ability to hydrolyze protein peptide bonds into amino acids. Bromelain enzyme can be found in pineapple (Ananas comosus) which consists of 95% cysteine protease mixture. Bromelain is often used by athletes to treat physical injuries or minor wounds. In obtaining the bromelain enzyme found in pineapple, an isolation process is required, namely by extracting it extracellularly. The method in this study does not go through the cell breakdown process so that it is easier to separate from other impurities and does not mix with other cell materials. This study aims to develop an innovative method in the production of bromelain enzyme using pineapple and honey as raw materials and to evaluate the organoleptic properties, pH, and the presence of lactic acid bacteria. This study is a descriptive study presented in the form of graphs and images. From the research that has been done, it can be concluded that the manufacture of bromelain enzyme using pineapple and honey has succeeded in producing a product that has an organoleptic value of liking (60%), pH 3 (acid) with the number of lactic acid bacteria 2.43 cells/ml.

Plant P450 forms indigo and indirubin when expressed in Escherichia coli

Indigo and indirubin are derived from indoxyl molecules, which generally occur as indoxyl glycosides in woad (Isatis tinctoria L.) and other indigo-producing plants. Indoxyl glycosides are biosynthesized from indole via 3-hydroxylation to form indoxyl, followed by one or more glycosylations. Enzymes that attach and remove sugars to and from indoxyl have already been isolated and characterized, while enzymes that convert indole into indoxyl in plants have remained elusive, until the identification of P450s and flavin-containing monooxygenases that hydroxylate indole. A P450 gene from woad (named CYP71B102) was heterologously expressed in E. coli, resulting in the formation of indigo and indirubin, as well as isatin and 2-oxindole, which along with indoxyl are putative precursors of indirubin. The addition of either isatin or 2-oxindole to the recombinant E. coli reduced the levels of indigo and increased the amount of indirubin, whereas coexpression of CYP71B102 with isatin hydroxylase (which degrades isatin) increased the levels of indigo and decreased the amount of indirubin, albeit slightly. The results suggest that CYP71B102 hydroxylates indole at both the 2- and 3- positions to produce 2-oxindole and indoxyl, respectively, and that the coupling of indoxyl with either 2-oxindole or isatin forms indirubin, while dimerization of indoxyl forms indigo. This P450 gene is thus likely involved in the biosynthesis of indirubin in woad, as well as the formation of indigo and its glycosidic precursors, even if other types of enzymes, such as flavin-containing monooxygenases, may be involved in indole hydroxylation in other indigo-producing plants.

Production of Citric Acid from Different Aspergillus species Obtained from Soil

The genus Aspergillus is considered highly important in the production of various types of enzymes and organic acids. Aspergillus species produce organic acids such as citric acid, itaconic acid, and malic acid, which are one of the most important alternate techniques for chemical processes. Citric acid is an essential primary agricultural product that is extensively used in the world. It acts as an intermediate in the tricarboxylic acid cycle (citric acid cycle) during the oxidation of carbohydrates to carbon dioxide. Citric acid has an important role in industries involving the production of food products, beverages, pharmaceutical and agricultural products, detergents, and cosmetics. The production of citric acid, a crucial agricultural commodity with widespread applications in various industries including food, beverages, pharmaceuticals, agriculture, detergents, and cosmetics, heavily relies on microbial fermentation. The great demand for citric acid due to its wide industrial applications and less toxicity. Citric acid can be produced using less expensive substrates that are renewable too. In this work, Aspergillus species was isolated from the soil source, collected in different localities of Chhatrapati Sambhajinagar. The total 90 different isolates are isolated among these the 20 different Aspergillus species are isolated and pure culture by using potato dextrose agar (PDA) Czapek dox agar medium and study the effect of different media on growth and sporulation of different Aspergillus species and the quantitative analysis of citric acid production was done by titration method with help of NaOH and phenolphthalein indicator. The main reason for the study increases is the large number of applications that can be found for citric acid, primarily in the food and pharmaceutical industries

Factors Affecting Cell Viability during the Enzymatic Dissociation of Human Endocrine Tumor Tissues.

The enzymatic dissociation of human solid tissues is a critical process for disaggregating extracellular matrix and the isolation of individual cells for various applications, including the immortalizing primary cells, creating novel cell lines, and performing flow cytometry and its specialized type, FACS, as well as conducting scRNA-seq studies. Tissue dissociation procedures should yield intact, highly viable single cells that preserve morphology and cell surface markers. However, endocrine tissues, such as adrenal gland tumors, thyroid carcinomas, and pituitary neuroendocrine tumors, present unique challenges due to their complex tissue organization and morphological features. Our study conducted a morphological examination of these tissues, highlighting the intricate structures and secondary degenerative changes that complicate the dissociation process. We investigated the effects of various dissociation parameters, including the types of enzymes, incubation duration, and post-dissociation purification procedures, such as debris removal and nontarget blood cell lysis, on the viability of cells derived from different tumor types. The findings emphasize the importance of optimizing tissue digestion protocols to preserve cell viability and integrity, ensuring reliable outcomes for downstream analyses.

Trichoderma role as an essential plant growth promotion-a review

In the world, pest infestation is responsible for 70% or more of per cent losses in agricultural productivity, and mycopesticide can provide an alternative option to pest control option as an alternative to synthetic pesticides. Trichoderma is a genus of fungal species, and Trichoderma viride is one of the known fungal species in categories that exhibit biocontrol mechanisms since it is well-identified for 100% effectiveness in pest control. The mycelium of T. viride has shown its capabilities for the production of various types of enzymes, including cellulases and chitinase, with degradation capability for celluloses and chitin polymers, respectively. It is ubiquitous in nature. It is widely used for the management of crop diseases and is basically soil-borne, but it is also foliar. Trichoderma has shown its potential due to diversity, ecology, and application. The mechanism of action includes competition, production of different metabolites, induced resistance and enzymes, etc. There are several commercial formulations available all over the world that are used against crop diseases. Trichoderma, due to its potential for degradation capability, can be utilised from waste to wealth. Later on, with the development of more research on Trichoderma, various utilities were found. This includes its effective control against harmful insects in agricultural as well as health sectors. It also can detoxify the heavy metals. In the health sector, continuous exploration of its benefits for drug discovery is being done. As a whole, it can be considered soil gold and a promising future beneficial microbe.

The genome of Eleocharis vivipara elucidates the genetics of C3-C4 photosynthetic plasticity and karyotype evolution in the Cyperaceae.

Eleocharis vivipara, an amphibious sedge in the Cyperaceae family, has several remarkable properties, most notably its alternate use of C3 photosynthesis underwater and C4 photosynthesis on land. However, the absence of genomic data has hindered its utility for evolutionary and genetic research. Here, we present a high-quality genome for E. vivipara, representing the first chromosome-level genome for the Eleocharis genus, with an approximate size of 965.22 Mb mainly distributed across 10 chromosomes. Its Hi-C pattern, chromosome clustering results, and one-to-one genome synteny across two subgroups indicates a tetraploid structure with chromosome count 2n = 4x = 20. Phylogenetic analysis suggests that E. vivipara diverged from Cyperus esculentus approximately 32.96 million years ago (Mya), and underwent a whole-genome duplication (WGD) about 3.5 Mya. Numerous fusion and fission events were identified between the chromosomes of E. vivipara and its close relatives. We demonstrate that E. vivipara has holocentromeres, a chromosomal feature which can maintain the stability of such chromosomal rearrangements. Experimental transplantation and cross-section studies showed its terrestrial culms developed C4 Kranz anatomy with increased number of chloroplasts in the bundle sheath (BS) cells. Gene expression and weighted gene co-expression network analysis (WGCNA) showed overall elevated expression of core genes associated with the C4 pathway, and significant enrichment of genes related to modified culm anatomy and photosynthesis efficiency. We found evidence of mixed nicotinamide adenine dinucleotide - malic enzyme and phosphoenolpyruvate carboxykinase type C4 photosynthesis in E. vivipara, and hypothesize that the evolution of C4 photosynthesis predates the WGD event. The mixed type is dominated by subgenome A and supplemented by subgenome B. Collectively, our findings not only shed light on the evolution of E. vivipara and karyotype within the Cyperaceae family, but also provide valuable insights into the transition between C3 and C4 photosynthesis, offering promising avenues for crop improvement and breeding.

Functional Peptides from Yak Milk Casein: Biological Activities and Structural Characteristics.

The average content of casein in yak milk is 40.2 g/L. Casein can be degraded by enzymatic digestion or food processing to produce abundant degradation peptides. International researchers have studied the degradation peptides of yak milk casein by using multiple techniques and methods, such as in vitro activity tests, cellular experiments, proteomics, bioinformatics, etc., and found that the degradation peptides have a wide range of functional activities that are beneficial to the human body, such as angiotensin-converting enzyme (ACE) inhibitory, antioxidant, anti-inflammatory, antidiabetic, antimicrobial, anticancer, and immunomodulatory activities, etc., and it has been proved that the types and strengths of functional activities are closely related to the structural characteristics of the peptides. This paper describes the characteristics of yak milk proteins, the functional activities, and mechanism of action of degraded peptides. Based on the types of functional activities of yak milk casein degradation peptides, we classified and elucidated the effects of structural factors, such as peptide molecular weight, peptide length, amino acid sequence, physicochemical properties, electrical charge, hydrophobicity, spatial conformation, chain length, and the type of enzyme on these activities. It reveals the great potential of yak milk casein degradation peptides as functional active peptide resources and as auxiliary treatments for diseases. It also provides important insights for analyzing yak casein degradation peptide activity and exploring high-value utilization.

Exploration and application prospect of the advanced technology of time–temperature indicators

The time–temperature indicator (TTI) is a kind of indicator that can monitor the safety of food, drugs and other products. It produces the cumulative effect of time–temperature through physical and mechanical deformation, chemical or biological reaction, and makes an irreversible and obvious color change, so as to record the temperature change course experienced by products. This review details the selection and characteristics of TTI materials in the last 15 years. The classification of TTIs, such as enzyme type, chemical type, microbial type, diffusion type, nanomaterial type and Maillard type, self-healing type and electronic type, are described in detail. Finally, the application of TTIs in various kinds of food packaging, such as meat products, aquatic products, fruits and vegetables and dairy products, is emphatically studied. This review provides ideas for the further improvement and research of TTIs, and provides a reference for the performance of TTIs in other commercial fields.

Dual β-lactams for the treatment of Mycobacterium abscessus: a review of the evidence and a call to act against an antibiotic nightmare.

Mycobacterium abscessus complex is a group of rapidly growing non-tuberculous mycobacteria (NTM), increasingly emerging as opportunistic pathogens. Current treatment options for these microorganisms are limited and associated with a high rate of treatment failure, toxicity and recurrence. In search of new therapeutic strategies, interest has grown in dual β-lactam (DBL) therapy, as research recently discovered that M. abscessus cell wall synthesis is mainly regulated by two types of enzymes (d,d-transpeptidases and l,d-transpeptidases) differently susceptible to inhibition by distinct β-lactams. In vitro studies testing several DBL combinations have shown synergy in extracellular broth cultures as well as in the intracellular setting: cefoxitin/imipenem, ceftaroline/imipenem, ceftazidime/ceftaroline and ceftazidime/imipenem. The addition of specific β-lactamase inhibitors (BLIs) targeting M. abscessus β-lactamase did not significantly enhance the activity of DBL combinations. However, in vivo data are lacking. We reviewed the literature on DBL/DBL-BLI-based therapies for M. abscessus infections to raise greater attention on this promising yet overlooked treatment option and to guide future preclinical and clinical studies.

Efficacy of an AAV vector encoding a thermostable form of glucocerebrosidase in alleviating symptoms in a Gaucher disease mouse model

Almost all attempts to date at gene therapy approaches for monogenetic disease have used the amino acid sequences of the natural protein. In the current study, we use a designed, thermostable form of glucocerebrosidase (GCase), the enzyme defective in Gaucher disease (GD), to attempt to alleviate neurological symptoms in a GD mouse that models type 3 disease, i.e. the chronic neuronopathic juvenile subtype. Upon injection of an AAVrh10 (adeno-associated virus, serotype rh10) vector containing the designed GCase (dGCase) into the left lateral ventricle of Gba−/−;Gbatg mice, a significant improvement in body weight and life-span was observed, compared to injection of the same mouse with the wild type enzyme (wtGCase). Moreover, a reduction in levels of glucosylceramide (GlcCer), and an increase in levels of GCase activity were seen in the right hemisphere of Gba−/−;Gbatg mice, concomitantly with a significant improvement in motor function, reduction of neuroinflammation and a reduction in mRNA levels of various genes shown previously to be elevated in the brain of mouse models of neurological forms of GD. Together, these data pave the way for the possible use of modified proteins in gene therapy for lysosomal storage diseases and other monogenetic disorders.

Workflow for development of CAPS markers with one type of restriction enzyme to identify citrus cultivars

Given the ease of propagating fruit tree species through cloning, the economic viability of their breeding programs hinges on protecting breeders' rights. This necessitates the development of highly accurate DNA markers for cultivar identification. Here, we present a methodology for the rapid design of cleaved amplified polymorphic sequence (CAPS) markers to discriminate newly bred Japanese citrus cultivars from genetically related cultivars. We first compared the performance of ddRAD-seq and MIG-seq in citrus germplasm. The ddRAD-seq libraries generated using EcoRI and HindIII restriction enzymes yielded the highest number of polymorphisms. Subsequently, ddRAD-seq with EcoRI and HindIII was employed to analyze 29 citrus cultivars and thus identify 331,801 genome-wide polymorphisms. A semi-automated bioinformatics pipeline was then utilized to identify candidate CAPS markers, resulting in the discovery of 14,072 potential markers. Of these candidates, 52 were chosen for validation based on their recognition by the PstI restriction enzyme. This evaluation resulted in the development of 11 highly discriminative CAPS markers. Remarkably, a combination of only six such markers was sufficient to differentiate newly bred cultivars from their genetically related parents. The single restriction enzyme employed for these markers facilitates straightforward multiplexing. Finally, a combination of one multiplex marker testing two loci and four singleplex markers was successfully selected that completely discriminated the cultivars other than the bud sports used in this study. The pipeline established here extends beyond citrus and has the potential to simplify marker development and cultivar protection in various plant species.

A study of the correlation between phospholipase A2 enzyme activity and anti-complement antibodies in patients with systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is an autoimmune disease that affects multiple organs or organ systems. SLE and one of its most common complications, lupus nephritis (LN), are characterized by chronic inflammation due to the secretion of pro-inflammatory molecules and a tissue deposition of immune complexes formed by autoantibodies and their target antigens, such as double-stranded DNA, other nuclear antigens, and complement proteins. Increased activity of phospholipase A2 enzymes (PLA2) like calcium-independent PLA2 (iPLA2), secretory PLA2 (sPLA2), and cellular PLA2 (cPLA2) has a crucial role in the maintenance of the inflammatory process during the course of SLE. This study aimed to evaluate PLA2 activity and identify the dominant type of PLA2 enzymes in a cohort of Bulgarian patients with SLE and LN. Additionally, we investigated whether the increased PLA2 activity was correlated with the presence of autoantibodies specific to the complement proteins C3 and factor H.

“The effects of non-surgical periodontal treatment plus zinc and magnesium supplementation on oxidative stress and antioxidants enzymes in type 2 diabetes patients: a quasi-experimental study”

BackgroundPeriodontal Disease (PD) associated with Type 2 Diabetes Mellitus (T2DM) is a chronic condition that affects the oral cavity of people living with T2DM. The mechanisms of the interaction between type 2 Diabetes Mellitus and Periodontal diseases are complex and involve multiple pathophysiological pathways related to the systemic inflammatory process and oxidative stress. Non-surgical periodontal treatment (NSTP) is considered the standard for the management of this disease; however, patients with systemic conditions such as type 2 Diabetes Mellitus do not seem to respond adequately. For this reason, the use of complementary treatments has been suggested to support non-surgical periodontal treatment to reduce the clinical consequences of the disease and improve the systemic conditions of the patient. The use of zinc gluconate and magnesium oxide as an adjunct to non-surgical periodontal treatment and its effects on periodontal clinical features and oxidative stress in patients with Periodontal diseases -type 2 Diabetes Mellitus is poorly understood.MethodsA quasi-experimental study was performed in patients with periodontal diseases associated with T2DM. Initially, 45 subjects who met the selection criteria were included. 19 were assigned to a control group [non-surgical periodontal treatment] and 20 to the experimental group (non-surgical periodontal treatment + 500 mg of magnesium oxide and 50 mg of zinc gluconate for oral supplementation for 30 days) and the data of 6 patients were eliminated. Sociodemographic characteristics, physiological factors, biochemical parameters, and clinical features of periodontal diseases were assessed.ResultsIn this research a change in periodontal clinical characteristics was observed, which has been associated with disease remission. Additionally, a shift in MDA levels was presented for both groups. Furthermore, the supplementation group showed an increase in antioxidant enzymes when compared to the group that only received NSPT.ConclusionThe use of Zinc gluconate and magnesium oxide can serve as a complementary treatment to non-surgical periodontal treatment, that supports the remission of PD as a result of regulation-reduction of oxidative biomarkers and increase in antioxidant enzymes activity.Trial Registrationhttps://www.isrctn.com ISRCTN 14,092,381. September 13º 2023. Retrospective Registration.Graphical

Differential effects of montelukast and zafirlukast on MDA‑MB‑231 triple‑negative breast cancer cells: Cell cycle regulation, apoptosis, autophagy, DNA damage and endoplasmic reticulum stress.

Montelukast and zafirlukast, cysteinyl leukotriene receptor antagonists (LTRAs), trigger apoptosis and inhibit cell proliferation of triple‑negative breast cancer MDA‑MB‑231 cells. By contrast, only zafirlukast induces G0/G1 cell cycle arrest. The present study compared the effects of these drugs on proteins regulating cell proliferation, apoptosis, autophagy, and endoplasmic reticulum (ER) and oxidative stress using reverse transcription‑quantitative PCR, western blotting and flow cytometry. The expression of proliferating markers, Ki‑67 and proliferating cell nuclear antigen, was decreased by both drugs. Zafirlukast, but not montelukast, decreased the expression of cyclin D1 and CDK4, disrupting progression from G1 to S phase. Zafirlukast also increased the expression of p27, a cell cycle inhibitor. Both drugs decreased the expression of anti‑apoptotic protein Bcl‑2 and ERK1/2 phosphorylation, and increased levels of the autophagy marker LC3‑II and DNA damage markers, including cleaved PARP‑1, phosphorylated (p)‑ATM and p‑histone H2AX. The number of caspase 3/7‑positive cells was greater in montelukast‑treated cells compared with zafirlukast‑treated cells. Montelukast induced higher levels of the ER stress marker CHOP compared with zafirlukast. Montelukast activated PERK, activating transcription factor 6 (ATF6) and inositol‑requiring enzyme type 1 (IRE1) pathways, while zafirlukast only stimulated ATF6 and IRE1 pathways. GSK2606414, a PERK inhibitor, decreased apoptosis mediated by montelukast, but did not affect zafirlukast‑induced cell death. The knockdown of CHOP by small interfering RNA reduced apoptosis triggered by montelukast and zafirlukast. In conclusion, the effects on cell cycle regulator proteins may contribute to cell cycle arrest caused by zafirlukast. The greater apoptotic effects of montelukast may be caused by the higher levels of activated caspase enzymes and the activation of three pathways of ER stress: PERK, ATF6, and IRE1.