6 research outputs found

    Phytocompounds-based therapeutic approach: Investigating curcumin and green tea extracts on MCF-7 breast cancer cell line

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    Background: Breast cancer (BC) has transcended lung cancer as the most common cancer in the world. Due to the disease's aggressiveness, rapid growth, and heterogeneity, it is crucial to investigate different therapeutic approaches for treatment. According to the World Health Organization (WHO), Plant-based therapeutics continue to be utilized as safe/non-toxic complementary or alternative treatments for cancer, even in developed countries, regardless of how cutting-edge conventional therapies are. Despite their low bioavailability, curcumin (CUR) and green tea (GT) represent safer therapeutic options. Due to their potent molecular-modulating properties on various cancer-related molecules and signaling pathways, they are considered gold-standard therapeutic agents and have been incorporated into the development of one or more therapeutic strategies of BC treatment. Methods: We investigated the modulatory role of CUR and GT extracts on significant multi molecular targets in MCF-7 BC cell line to assess their potential as BC multi-targeting agents. We analyzed the phytocompounds in GT leaves using High-performance liquid chromatography (HPLC) and Gas chromatography-mass spectrometry (GC-MS) techniques. The mRNA expression levels of Raf-1, Telomerase, Tumor necrosis factor alpha (TNF-α) and Interleukin-8 (IL-8) genes in MCF-7 cells were quantified using quantitative real-time PCR (qRT-PCR). The cytotoxicity of the extracts was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the released Lactate dehydrogenase (LDH), a valuable marker for identifying the programmed necrosis (necroptosis). Additionally, the concentrations of the necroptosis-related proinflammatory cytokines (TNF-α and IL-8) were measured using enzyme-linked immunosorbent assay (ELISA). Results: In contrast to the GT, the results showed the anticancer and cytotoxic properties of CUR against MCF-7 cells, with a relatively higher level of released LDH. The CUR extract downregulated the oncogenic Raf-1, suppressed the Telomerase and upregulated the TNF-α and IL-8 genes. Results from the ELISA showed a notable increase in IL-8 and TNF-α cytokines levels after CUR treatment, which culminated after 72 h. Conclusions: Among both extracts, only CUR effectively modulated the understudy molecular targets, achieving multi-targeting anticancer activity against MCF-7 cells. Moreover, the applied dosage significantly increased levels of the proinflammatory cytokines, which represent a component of the cytokines-targeting-based therapeutic strategy. However, further investigations are recommended to validate this therapeutic approach

    Genetic Diversity of Selected Rice Genotypes under Water Stress Conditions

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    Drought is the most challenging abiotic stress for rice production in the world. Thus, developing new rice genotype tolerance to water scarcity is one of the best strategies to achieve and maximize high yield potential with water savings. The study aims to characterize 16 rice genotypes for grain and agronomic parameters under normal and drought stress conditions, and genetic differentiation, by determining specific DNA markers related to drought tolerance using Simple Sequence Repeats (SSR) markers and grouping cultivars, establishing their genetic relationship for different traits. The experiment was conducted under irrigated (normal) and water stress conditions. Mean squares due to genotype × environment interactions were highly significant for major traits. For the number of panicles/plants, the genotypes Giza179, IET1444, Hybrid1, and Hybrid2 showed the maximum mean values. The required sterility percentage values were produced by genotypes IET1444, Giza178, Hybrid2, and Giza179, while, Sakha101, Giza179, Hybrid1, and Hybrid2 achieved the highest values of grain yield/plant. The genotypes Giza178, Giza179, Hybrid1, and Hybrid2, produced maximum values for water use efficiency. The effective number of alleles per locus ranged from 1.20 alleles to 3.0 alleles with an average of 1.28 alleles, and the He values for all SSR markers used varied from 0.94 to 1.00 with an average of 0.98. The polymorphic information content (PIC) values for the SSR were varied from 0.83 to 0.99, with an average of 0.95 along with a highly significant correlation between PIC values and the number of amplified alleles detected per locus. The highest similarity coefficient between Giza181 and Giza182 (Indica type) was observed and are susceptible to drought stress. High similarity percentage between the genotypes (japonica type; Sakha104 with Sakha102 and Sakha106 (0.45), Sakha101 with Sakha102 and Sakha106 (0.40), Sakha105 with Hybrid1 (0.40), Hybrid1 with Giza178 (0.40) and GZ1368-S-5-4 with Giza181 (0.40)) was also observed, which are also susceptible to drought stress. All genotypes are grouped into two major clusters in the dendrogram at 66% similarity based on Jaccard’s similarity index. The first cluster (A) was divided into two minor groups A1 and A2, in which A1 had two groups A1-1 and A1-2, containing drought-tolerant genotypes like IET1444, GZ1386-S-5-4 and Hybrid1. On the other hand, the A1-2 cluster divided into A1-2-1 containing Hybrid2 genotype and A1-2-2 containing Giza179 and Giza178 at coefficient 0.91, showing moderate tolerance to drought stress. The genotypes GZ1368-S-5-4, IET1444, Giza 178, and Giza179, could be included as appropriate materials for developing a drought-tolerant variety breeding program. Genetic diversity to grow new rice cultivars that combine drought tolerance with high grain yields is essential to maintaining food security

    MOLECULAR EPIDEMIOLOGICAL STUDY ON PESTE DES PETITS RUMINANTS IN EGYPT 2015

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    Peste des petits ruminants (PPR) is one of highly contagious viral diseases of small ruminants with high economic losses due to the high morbidity and mortality. In Egypt, PPR in the last 10 years re-emerged again with high mortality in sheep flocks. There is no much data regarding the circulating Peste des petits ruminants’ virus and the epidemiological distribution in small ruminants. The study was carried out on a sheep flock of 50 rams aged of 9-12 months with clinical signs suggestive to PPR infection (fever, erosions of the buccal mucosa, pneumonia, diarrhea high morbidity and mortality). Buffy coat and lymph nodes from diseased and dead animals were sent for diagnosis and molecular diagnosis was confirmed using RT-PCR with specific primers targeting three genes; nucleo-capsid (N), fusion (F), and hemagglutinin (H). Sequencing and phylogenetic analysis was carried out on the PCR products and revealed that, the circulating virus is belonged to lineage IV along with Ethiopian strain.Key words: peste des petits ruminants; molecular; epidemiological; diagnosis; Egypt MOLEKULARNA EPIDEMIOLOŠKA ŠTUDIJA KUGE DROBNICE V EGIPTU LETA 2015Povzetek: Kuga drobnice (PPR; iz angl. peste des petits) je ena izmed zelo nalezljivih virusnih bolezni malih prežvekovalcev, ki zaradi visoke obolevnosti in umrljivosti povzroča visoke gospodarske izgube. V Egiptu se je v zadnjih desetih letih PPR znova pojavila z visoko smrtnostjo pri ovcah. O kugi drobnice in epidemiologiji te bolezni pri malih prežvekovalcih v literaturi ne najdemo veliko podatkov. Opisana raziskava je bila opravljena v čredi 50 ovnov, starih od 9 do 12 mesecev, s kliničnimi znaki, ki so kazali na prisotnost bolezni PPR (zvišana telesna temperatura, erozije ustne sluznice, pljučnica, driska, visoka obolevnost in smrtnost). Za natančno diagnosticiranje bolezni je bil uporabljen del krvi, pridobljen s centrifugiranjem, ki vsebuje veliko levkocitov in trombocitov (angl. Buffy coat) in bezgavke obolelih in mrtvih ovnov. Z uporabo specifičnih oligonukleotidov za določanje prisotnosti nukleotidne (N), fuzijske (F) in hemaglutininske (H). beljakovine v reakciji RT PCR smo na molekularnem nivoju potrdili prisotnost okužbe s povzročiteljem PPR. Analiza zaporedja genov in filogenetske analize so bile izvedene na produktih analize PCR, pri tem pa smo ugotovili, da virus pripada liniji IV skupaj z etiopijskim sevom.Ključne besede: kuga drobnice; molekularna diagnostika; epidemiologija; Egip

    Genetic diversity and antibiotic resistance of Mycobacterium bovis in bovines in the Delta area of Egypt

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    IntroductionMycobacterium bovis (M. bovis) causes significant financial harm to the cattle industry through decreased productivity and trade limitations, while also posing a risk to human health through zoonotic transmission, which is primarily from unpasteurized milk or close animal contact.MethodsSingle intradermal tuberculin was used to test 2400 cases (1000 Holstein Friesian cattle and 1400 native breed buffaloes) during the national control program from Cairo, El-Buhaira, Dakahlia, Gharbia, Menoufia, and Sharkia districts located at the northern areas of Egypt. Tuberculin-positive cases were slaughtered and subjected to postmortem examination and isolation of M. bovis was performed. IS6110 primer was used in PCR test to confirm the existence of genus mycobacterium and regions of difference-based differentiation was used to detect M. bovis on the species level, phenotypic and genotypic antimicrobial resistance, as well as mycobacterial interspersed repetitive unit-variable number tandem repeat analysis (MIRU-VNTR) were performed.ResultsA total of 65 out of 2400 (2.7%) cases were single intradermal tuberculin test positive, 40 out of 65 (61.53%) were M. bovis positive on PCR, and the 40 isolates exhibited susceptibility to ethambutol, rifampicin, streptomycin, ciprofloxacin, levofloxacin, ofloxacin, and sparfloxacin. From them, 32 (80%) were susceptible to isoniazid, and 8 (20%) were resistant. These eight isolates contained three distinct katG mutations at codons 315, 463, and 506 with rates of 2/8 (25%), 3/8 (37.5%), and 3/8 (37.5%), respectively each representing a unique, single-codon mutation. MIRU-VNTR analysis enabled the identification of 35 distinct genotypes, with genotypes 26, 27, and 28 showing high prevalence. The nine highly discriminatory loci MIRU10, QUB11b, MIRU26, QUB26, QUB4156, MIRU04 ETRD, ETRA, Mtub30, and Mtub39 with a discriminating index of 0.9676 were suitable for the preliminary genotyping of M. bovis isolates from animals. M. bovis, ID: 7540/01, Lineage: Bovis and ID: 951/01, Lineage: Bovis from Germany were the closest lineages to our genotypes using the MIRU-VNTR plus database.ConclusionM. bovis isolated from cattle and buffaloes of some Delta area districts expressed high diversity and some isolates showed resistance to isoniazid with katG mutations. Continuous implementation of MIRU-VNTR analysis will help to trace the origin and similarities among animal and human isolates within the Delta area

    Phloretin-nanospanlastics for targeting the Akt/PI3K signaling pathways in dimethylhydrazine-induced colon cancer in mice

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    Objectives: Colorectal cancer is the third most common cancer worldwide, accounting for approximately 10 % of all cancer cases. It is also the second leading cause of cancer-related deaths globally. Phloretin is a natural compound found in apples and other fruits. It has been studied for its potential health benefits, including antioxidant and anti-inflammatory properties. However, more research is needed to fully understand its impact on cancer prevention or treatment. This article aimed to prepare phloretin-nanospanlastics (Ph-NSLs) to evaluate their effects on dimethylhydrazine (DMH)-induced colon cancer in mice. Methods: Morphology, Particle size, zeta potential, UV–vis, entrapment efficiency, polydispersity index, FT-IR spectra, and drug release of phloretin and Ph-NSLs at pH 6.8.were described. Ph-NSLs were also tested for their anti-cancer properties in DMH-induced colon cancer in mice. A 36 mice were divided into 6 groups; Normal control, DMH (20 mg/k.g.b.w.), DMH + Ph-NSLs (25 mg/k.g.b.w.), DMH + Ph-NSLs (50 mg/k.g.b.w.), DMH + 5-FU(20 mg/k.g.b.w.), DMH + Ph-NSLs (50 mg), 5-FU (20 mg). Ph-NSLs were tested for their anticancer properties in DMH-treated mice by evaluating the IC50, viability and inhibitory values of Ph-NSLs against Caco-2. Also, the effect of Ph-NSLs administration on number of surviving mice, number of tumors/mice, average of tumor size, Hb, RBCs, WBCs, C19–9, MDA, GSH, SOD, IL-2, TNF-α, TGF-β1, CEA, and P53 levels in mice treated DMH were estimated. Results: The synthesized Ph-NSLs were uniform, spherically shaped, and well dispersed, with a size, entrapment efficiency, and polydispersity index of approximately 114.06 ± 8.35 nm, 78.60 %, and 0.05, respectively. The zeta potential value of Ph-NSLs was measured at −21.5 ± 1.47 mV. Zeta potential reflects the surface charge of nanoparticles and affects their stability and interactions. UV spectra of phloretin and Ph-NSLs showed strong absorption peaks at 225 and 285 nm. These peaks correspond to specific wavelengths where the compound absorbs light. The percentage of Ph- NSLs release was found to be 56.87 ± 2.45 %. IC50 of Ph-NSLs was recorded 15.76 ± 0.42 μg/ml and the viability and inhibitory values of Ph-NSLs against Caco-2 cell lines was resorded 2.39, and 97.61 %, respectively at 100 μg/ml as well as 10.3, and 89.7 %, respectively at 50 μg/ml.Moreover, The combination of 5-FU and Ph-NSLs resulted in a moderate increase in survival and significantly reduces tumor size and number, showing enhanced anticancer efficacy compared to individual treatments as well as attenuated levels of hemoglobin (Hb), red blood cells (RBCs), and white blood cells (WBCs). Reduced plasma cancer antigen 19–9 (CA19–9) levels as well as improved of colon malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), interleukine-2 (IL-2), tumor necrosis factor-alpha (TNF-α), tumor growth factor-beta1 (TGF-β1), carcinoembryonic antigen (CEA), and tumor protein (P53) levels. Also, Ph-NSLs and 5FU, either alone or together, decreased the expression of the Akt and PI3K genes in the colon. The combination of Ph-NSLs and 5FU showed more pronounced anticancer activity than Ph-NSLs administered individually. Conclusion: The combination of 5-FU and Ph-NSLs significantly enhances anticancer efficacy, reducing both the number of tumors and average tumor size more effectively than either treatment alone. This synergistic effect leverages 5-FU's inhibition of DNA synthesis and phloretin's induction of apoptosis and inhibition of cell proliferation, offering a promising approach for improved cancer treatment outcomes

    Exploring agro-morphological and fiber traits diversity in cotton (G. barbadense L.)

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    Abstract Cotton (Gossypium barbadense L.) is a leading fiber and oilseed crop globally, but genetic diversity among breeding materials is often limited. This study analyzed genetic variability in 14 cotton genotypes from Egypt and other countries, including both cultivated varieties and wild types, using agro-morphological traits and genomic SSR markers. Field experiments were conducted over two seasons to evaluate 12 key traits related to plant growth, yield components, and fiber quality. Molecular diversity analysis utilized 10 SSR primers to generate DNA profiles. The Molecular diversity analysis utilized 10 SSR primers to generate DNA profiles. Data showed wide variation for the morphological traits, with Egyptian genotypes generally exhibiting higher means for vegetative growth and yield parameters. The top-performing genotypes for yield were Giza 96, Giza 94, and Big Black Boll genotypes, while Giza 96, Giza 92, and Giza 70 ranked highest for fiber length, strength, and fineness. In contrast, molecular profiles were highly polymorphic across all genotypes, including 82.5% polymorphic bands out of 212. Polymorphism information content was high for the SSR markers, ranging from 0.76 to 0.86. Genetic similarity coefficients based on the SSR data varied extensively from 0.58 to 0.91, and cluster analysis separated genotypes into two major groups according to geographical origin. The cotton genotypes displayed high diversity in morphology and genetics, indicating sufficient variability in the germplasm. The combined use of physical traits and molecular markers gave a thorough understanding of the genetic diversity and relationships between Egyptian and global cotton varieties. The SSR markers effectively profiled the genotypes and can help select ideal parents for enhancing cotton through hybridization and marker-assisted breeding
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