Formulation of quinoa oil-alginate loaded nanoemulsion and its anticancer efficacy as a therapy for chemically induced breast cancer.

BACKGROUND: Quinoa seeds (Chenopodium quinoa Willd.) have gained interest due to their naturally occurring phytochemicals and antioxidants. They possess potent anticancer properties against human colorectal cancer. METHODS AND RESULTS: Fatty acids in quinoa oil were studied using gas chromatography-mass spectrometry. Rats were used to test the acute oral toxicity of the nanoemulsion loaded with sodium alginate. The DPPH radical scavenging method was employed to assess the nanoemulsion's ability to scavenge free radicals. It was examined the in vivo anticancer potential of quinoa oil nanoemulsion on rats with breast cancer induced by 7, 12-dimethylbenz (a) anthracene (DMBA). DMBA-breast cancer models received daily quinoa oil nanoemulsions for 30 days. The anticancer effect of the nanoemulsion was assessed by measuring ROS, protein carbonyl, gene expression of anti-oncogenes, and histopathological analysis. Supplying quinoa oil nanoemulsion significantly reduced the increase in serum ROS and PC levels induced in breast cancer tissue. The expression levels of antioncogenes in breast cancer tissue were decreased by the quinoa oil nanoemulsion. Nanoemulsions also improved the cellular morphology of breast tumors. CONCLUSION: The study results indicate that quinoa oil nanoemulsion has anticancer activity against breast cancer, effectively modulating oxidative stress markers, anti-oncogene expressions, and tissue architecture. It can be inferred from the results that quinoa oil nanoemulsion is a chemoprotective medication that may hinder breast cancer progression in rats.

Genetic similarity and diversity among three camel populations reared in Egypt.

BACKGROUND: Molecular genetics has been extremely useful in determining the relation between animal populations and documenting the degrees of genetic variation found within them. The present study was undertaken to evaluate genetic diversity and the relationships between the three camel populations reared in Egypt: Maghrabi, Sudani, and Baladi using mitochondrial 16S sequences and other breeds of camels in the world. METHODS: Blood samples were collected from camels belonging to these three populations. Genomic DNA was extracted from the collected blood samples and subjected to PCR using specific primers for mitochondrial 16S region. The amplified products were purified using DNA purification kit to remove residual primers and dNTPs. Sequencing was performed in the Macrogen Incorporation. The amplified products were submitted to GenBank/NCBI under accession numbers OM 278349 and OM 278350 RESULTS: Sequencing was done on the partial mitochondrial 16S amplified fragments at 530 bp. This amplified area had two haplotypes. There was one substitution (G/A) at nucleotide 309 of the amplified segment. The nucleotide (π) and Hd stand for haplotype diversity, respectively, at 0.00008 and 0.042, and the average number of pairwise nucleotide differences, k, is 0.042, according to Fu's Fs statistic and Tajima's D, which is -1.10686. Genetic distance percentages between the three populations under study range from 0.000 to 0.0312. A phylogenetic analysis of Egyptian camel populations and other Camelus dromedarius populations revealed a strong relationship between them. CONCLUSIONS: This study suggests that the 16S rRNA sequencing in mitochondria plays a critical role in genetic variation studies and analysis of phylogeny between camel populations and breeds.

Evaluation of the effectiveness of two new strains of Lactobacillus on obesity-induced kidney diseases in BALB/c mice.

BACKGROUND: Kidney disease (KD) is a public health problem worldwide and is an important factor in peripheral vascular disease, arrhythmias, heart failure, acute myocardial infarction, stroke, and angina. Obesity has been indicated as an effective cause of kidney diseases. So, this study aims to use two new strains of Lactobacillus to reduce the metabolic disorders and kidney insufficiency associated with obesity. METHODS: Fifty BALB/c male mice were divided into five groups (control, obesity, obesity pro1, obesity pro2, and obesity mix). The bodyweight, cholesterol profile, urea, and creatinine levels in urine and serum were all measured. Histopathological analysis and expression of Opn, Vim, Ngal, Kim-1, and αKlotho genes for kidney tissues were performed. RESULTS: The results indicated that body weight, cholesterol profile, urea, and creatinine levels in serum and urine had the lowest significance (P ˂ 0.05) in the obesity mix group and the highest significance in the obesity group. HDL had the highest significance (P ˂ 0.05) in the obesity mix group and the lowest significance (P ˂ 0.05) in the obesity group. Expression of Opn, Vim, Ngal, and Kim-1 genes was the most upregulated in the obesity group compared with the other groups, and there were nonsignificant differences (P > 0.05) between the obesity pro1 and obesity mix groups and the control group. Expression of αKlotho gene was significantly reduced (P ˂ 0.05) in the obesity group compared with the control group. CONCLUSION: This study demonstrated that the combination of pro1 and pro2 strains could reduce kidney inflammation and necrosis.

The suppressive role of nanoencapsulated chia oil against DMBA-induced breast cancer through oxidative stress repression and tumor genes expression modulation in rats.

BACKGROUND: Chia oil is high in omega-3 fatty acids, which have been linked to a lower risk of many diseases, including cancer. Oil encapsulation is a method that holds promise for maintaining oil content while enhancing solubility and stability. The purpose of this study is to prepare nanoencapsulated Chia oil and investigate its suppressive effects on rat chemically induced breast cancer. METHODS: The oil was extracted from commercial Chia seeds and their fatty acids were analyzed using Gas Chromatography-mass spectrometry (GC/MS). Sodium alginate was used as a loading agent to create the Chia oil nanocapsules. The DPPH assay was used to assess the oil nanocapsules' capacity to scavenge free radicals. Breast cancer induction was done by single dose subcutaneously administration of 80 mg/kg dimethylbenz (a) anthracene (DMBA). Models of breast cancer were given Chia oil nanocapsules orally for one month at doses of 100 and 200 mg/kg. Through measuring intracellular reactive oxygen species (ROS) and protein carbonyl, assessing the gene expression of tumor suppressor genes (BRCA 1 & 2, TP53), and conducting histopathological analysis, the suppressive effect of Chia oil nanocapsules was examined. RESULTS: The increase in ROS and PC levels brought on by DMBA was significantly decreased by the administration of Chia oil nanocapsules. In tumor tissue from rats given Chia oil nanocapsules, the mRNA expression levels of BRCA1, BRCA2, and TP53 were controlled Histopathological analysis clarified that the tissue architecture of breast tumors was improved by nanocapsules management. CONCLUSIONS: These findings demonstrate the ability of Chia oil nanocapsules to inhibit cancer cells in the rat breast.

Topiramate potential neurotoxicity and mitigating role of ginger oil in mice brain.

Topiramate has multiple pharmacological mechanisms that are efficient in treating epilepsy and migraine. Ginger has been established to have gingerols and shogaols that cause migraine relief. Moreover, Topiramate has many off-label uses. Thus, it was necessary to explore the possible neurotoxicity of Topiramate and the role of ginger oil in attenuating the Topiramate neurotoxicity. Male albino mice were orally gavaged with Topiramate, ginger oil (400 mg/kg), and Topiramate plus ginger oil with the same pattern for 28 days. Oxidative stress markers, acetylcholinesterase (AchE), gamma-aminobutyric acid (GABA), and tumor necrosis factor-alpha (TNF-α) were examined. Histopathological examination, immunohistochemical glial fibrillary acidic protein (GFAP), and Bax expression analysis were detected. The GABAAR subunits, Gabra1, Gabra3, and Gabra5 expression, were assessed by RT-qPCR. The investigation showed that Topiramate raised oxidative stress markers levels, neurotransmitters, TNF-α, and diminished glutathione (GSH). In addition, Topiramate exhibited various neuropathological alterations, strong Bax, and GFAP immune-reactivity in the cerebral cortex. At the same time, the results indicated that ginger oil had no neurotoxicity. The effect of Topiramate plus ginger oil alleviated the changes induced by Topiramate in the tested parameters. Both Topiramate and ginger oil upregulated the mRNA expression of gabra1 and gabra3, while their interaction markedly downregulated them. Therefore, it could be concluded that the Topiramate overdose could cause neurotoxicity, but the interaction with ginger oil may reduce Topiramate-induced neurotoxicity and should be taken in parallel.

Genetic structure of some candidate genes of repeat breeder syndrome in Egyptian buffaloes.

BACKGROUND: This study aimed to explore the association between polymorphisms in three genes: leptin (LEP), leptin receptor (LEPR), and BMP4, and incidence of repeat breeding in Egyptian buffaloes. METHODS: DNA was extracted from 160 female buffaloes, involving 108 fertile and 52 repeat breeders. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Sequence analysis and alignment were performed by employing NCBI/BLAST/blastn suite, to identify SNPs among different patterns and alleles. We utilized PredictSNP software to predict the non-synonymous SNPs influences on protein function. Moreover, the conservation score of the amino acids within the target proteins was computed by ConSurf server. RESULTS: The genotyping results showed that LEP and BMP4 genes were monomorphic (CC, GG) in all tested fertile and repeat breeder buffaloes. Leptin gene sequencing showed a non-synonymous C73T SNP, replacing R to C at position 25 within the leptin polypeptide (position 4 in the mature form; R4C) which is a neutral mutation, not affecting function or structure of LEP protein. For LEPR, one synonymous SNP (T102C) and two non-synonymous SNPs (A106G and C146A), triggering V967A and G954C replacements, respectively in LEPR protein. Moreover, they are neutral mutations. Sequencing results of BMP4 showed HinfI restriction site indicate fixed GG genotype (CC genotype in the anti-sense strand) in all sequenced samples. No SNPs were observed within the amplified region. CONCLUSION: Genotyping and sequencing results of the surveyed three genes revealed that there is no association between these genes mutations and the incidence of repeat breeding in Egyptian buffaloes.

Genotoxic, biochemical and histopathological studies to assessment the topiramate hepatorenal toxicity in mice.

Liver and kidney role in detoxification and drug metabolism increases the risk of their poisonous injury. Topiramate (TMP) is an effective popular migraine prophylaxis that is accepted for utilize in adults and teenagers. Therefore, the target of this research is to estimate the potential toxic effects of TMP on liver and kidney in male mice. Thirty-two adult albino male mice were divided into four groups (n = 8 mice). Group I of animals was given saline solution and used as negative control. The other three groups were administrated TPM at doses (100, 200 and 400 mg/kg) for 28 days. Genotoxicity was evaluated by comet assay and DNA fragmentation by Diphenyleamine. Biochemical investigation was achieved by estimating liver enzymes (AST, ALT), alkaline phosphatase (ALP) creatinine and uric acid. In addition, measurement of the antioxidant enzymes, malondialdehyde and nitric oxide were performed in both two tissues of liver and kidney. Microscopic examination of hematoxyline and eosin (H&E), tumor necrosis factor (TNF-α) and caspase3 stained sections were done to explore the effect of topiramate on mice tissues of liver and kidney. The data revealed that TPM showed dose dependent toxicity that represented in: DNA damage in tested cells and increased level of liver enzymes, creatinine and uric acid as markers of toxicity. Topiramate significantly diminished antioxidant enzymes activities and elevated the level of malondialdehyde and nitric oxide. In addition, TPM caused histopathological alterations and dose dependent positive immune reaction for TNF--α and caspase 3 in kidney and liver tissues. The results showed that Topiramate has marked toxicity in liver and kidney of mice.

Molecular characterization and immunohistochemical localization of tilapia piscidin 3 in response to Aeromonas hydrophila infection in Nile tilapia.

The antimicrobial activity of tilapia piscidin 3 (TP3) was determined in vitro against a locally isolated Aeromonas hydrophila. A 388 bp fragment was amplified from the TP3 cDNA and sequenced. The coding sequence (CDS) of TP3 was estimated to be 231 bp codes for 76 amino acids long and stop codon. In silico analysis was performed to detect both the signal peptide and the prodomain cleavage sites to follow the amino acids number 22 and 70, respectively. Based on this, a peptide 23 amino acids long with a remarkably high computed antimicrobial probability was synthesized and used in the subsequent experiments. The antimicrobial activity of TP3 was determined with minimum inhibitory concentration (MIC) and minim um bactericidal concentration (MBC) methods. TP3 exhibited relatively weak antimicrobial activities against the tested bacteria. A challenge experiment was then performed in Nile tilapia with low and high doses of A. hydrophila, followed by timely recognition; after 3, 6, 24 h, and 7 days of the specific TP3 gene expression, immunohistochemical localization was also performed. Histopathological examination revealed provoked inflammatory responses and congestion in the same organs of TP3 expression. Immunohistochemical localization showed that A. hydrophila induced tilapia fish to express TP3 after 24 h within the gills, intestine, hepatopancreas, spleen, and posterior kidney. In quantitative real time (RT)-polymerase chain reaction analysis, the high dose showed higher mRNA expression levels than the low dose, and its expression levels increased in the A. hydrophila-infected fish. It was therefore concluded that TP3 plays an essential role in fish immunity.

Piscidin 4: Genetic expression and comparative immunolocalization in Nile tilapia (Oreochromis niloticus) following challenge using different local bacterial strains.

The antimicrobial activity of tilapia piscidin 4 (TP4) was determined in vitro against four bacterial strains, Aeromonas hydrophilla, Pseudomonas fluorescens, Streptococcus iniae and Vibrio anguillarum. Nile tilapia were infected with low and high doses of the tested pathogens; after 3, 6, 24 h and 7 days of the specific TP4 gene expression, tissue immunolocalization was also performed. Histopathological examination revealed septicaemia and necrosis of hemopoietic tissue for all of the tested bacteria. Immunolocalization showed abundance in S. iniae-infected fish tissues. Quantitative RT-PCR analysis revealed that high doses raised mRNA expression levels compared to low doses and expression levels increased in the infected fish, particularly after 24 h, indicating that TP4 exerts potent bactericidal activity against some fish pathogens and plays an essential role in fish immunity.

Efficiency of turnip bioactive lipids in treating osteoporosis through activation of Osterix and suppression of Cathepsin K and TNF-α signaling in rats.

Vegetable oils are characterized by their bioactive phytochemicals including fatty acids, tocols, and phenolic compounds. In the current study, turnip (Brassica rapa) oil was evaluated for its fatty acid profiles, tocol composition, and total phenolic content. The radical scavenging properties of oil against DPPH· and galvinoxyl radicals were also evaluated. Turnip oil efficiency in treating osteoporosis was tested in rats. Fifty adult female Sprague-Dawley albino rats were divided to five groups (n = 10/group). An osteoporotic rat model was prepared by two separate 5-day (5 days on/9 days off) courses of methotrexate subcutaneous injection. Osteoporotic rats were orally gavaged with turnip oil (200 and 400 mg/kg/day) for 28 days. Turnip oil efficiency in treating osteoporosis was studied by evaluation of Osterix, Cath K, and TNF-α transcript expression levels that involved in bone remodeling in femoral bones. Minerals and vitamin D were estimated in blood serum. Femoral bone histological and morphometric analyses were investigated in osteoporotic and turnip oil-treated rats. In vitro assays revealed strong antiradical potential of turnip oil. Treatment with turnip oil regulated the levels of Osterix, Cath K, and TNF-α mRNA that was accompanied with elevating the levels of calcium, phosphorous, bone alkaline phosphatase (BALP), and vitamin D in osteoporotic rats. The histological and morphometric inspection revealed that turnip oil displayed progress in the osteoporotic rat bone formation that was clear in the enhancement of thickness of femur shaft cortical bone and femur head trabecular bone. Above-mentioned findings indicated that turnip oil has the potential to share in the treatment of osteoporosis.

Effect of antiepileptic drug (Topiramate) and cold pressed ginger oil on testicular genes expression, sexual hormones and histopathological alterations in mice.

Sexual dysfunction in the epileptic patient is difficult to confirm whether it is ailment or therapy related. Antiepileptic drugs often use in reproductive age, through reproductive progress and maturation. On the other side, cold-pressed oils are rich in bioactive phytochemicals with health-promoting traits. The target of this work was to appraise the sexual dysfunction of antiepileptic Topiramate (TPM) and cold pressed ginger oil (CPGO) as antiepileptic alternative medicine in male mice. Fifty-four adult male albino mice were divided into nine groups (n = 6 mice). One group given saline and used as negative control; another one was given corn oil as vehicle. Six groups administered orally with TPM or CPGO at 100, 200 and 400 mg/kg. Moreover, group of animals co-administrated orally CPGO with TPM (400 mg/kg) to study their interaction. Fatty acid profile and tocols composition of CPGO were determined. in vitro assays were undertaken to evaluate radical scavenging traits of CPGO utilizing sable 1,1-diphenyl-2-picrylhydrazyl (DPPH·) and galvinoxyl radicals. The study investigated antioxidant and oxidative stress markers, sexual hormones levels, mRNA levels of vascular endothelial growth factor (Vegfa), synaptonemal complex protein (Sycp3), Wilms tumor gene (Wt1) as well as histopathological and immunohistochemical examination. Strong radical scavenging potential of CPGO against stable DPPH· and galvinoxyl radicals was recorded. The results revealed that TPM caused a dose-dependent reduction in the antioxidant activities and testosterone content, while, malonaldehyde (MDA) and nitric oxide (NO) as oxidative stress markers were elevated. Vegfa and Sycp3 mRNA expression down-regulated at all Topiramate tested doses, but Wt1 up-regulated at 400 mg/kg. TPM (400 mg/kg) revealed histological alterations associated with strong positive Bax immune reactive spermatogoneal and Leydig cells. Ginger oil elevated the CAT and SOD (antioxidant enzymes), serum testosterone and diminished the oxidative stress, up regulated the expression of Vegfa and Sycp3 and down-regulated the Wt1 expression. Meanwhile, CPGO revealed no histopathological alterations and no Bax immune-reactive cells. CPGO co-administration with TPM (400 mg/kg) attenuated the TPM toxicity. High doses of TPM may exhibit sexual dysfunction but CPGO is safe and has androgenic property. CPGO co-administration could protect the antiepileptic patient from the TPM sexual dysfunction.

Serological and molecular diagnostic surveys combined with examining hematological profiles suggests increased levels of infection and hematological response of cattle to babesiosis infections compared to native buffaloes in Egypt.

BACKGROUND: Babesiosis threatens the development of the cattle and buffaloes industries in Egypt and improved control is needed. The main objectives of this study are surveying the presence of bovine babesiosis in distinct selected bovine and buffalo populations in Egypt using novel molecular and previously validated serological methods, while also comparing the occurrence of hematological alterations among Babesia infected cattle and buffalos. METHODS: A total of 253 and 81 blood samples from apparently healthy cattle and buffaloes, respectively, were randomly collected from diverse locations in Egypt. All samples were tested for Babesia bovis and B. bigemina infection using blood film examination, competitive ELISA (cELISA) and PCR. Novel semi-nested and nested PCR assays for the detection of B. bovis and B. bigemina respectively, were developed and used to analyze DNA extracted from bovine and buffalo samples. Hematological profiles were studied using a hematological analyzer. RESULTS: Blood films examination revealed 13.8% and 7.4% Babesia infection rates in cattle and buffaloes, respectively. However, in cattle, the cELISA detected 32.8%, 21.3% and 10.7% infection rates with B. bigemina, B. bovis and mixed infection, respectively. In addition, cELISA identified 22.2%, 22.2% and 6.2% infection rates with B. bigemina, B. bovis and mixed infection, respectively in buffaloes. The semi-nested PCR assay showed that 15% of the tested samples were positive for B. bovis in cattle, but just 3% in buffaloes. Infections with B. bigemina were also found in cattle (32.4%), but not in buffaloes upon nested PCR analysis. Sequencing analysis confirmed the identity of the PCR amplicons and showed that Egyptian genotypes of B. bigemina and B. bovis highly resemble sequences previously deposited in GenBank. Hemograms performed on the sampled animals revealed macrocytic hypochromic anemia associated with reduced platelet counts in infected cattle with babesiosis. In addition, marked increases in total leukocyte and granulocytic counts and decreases in lymphocytic counts were found in infected cattle. In contrast, no such hematological anomalies were found in presumably Babesia-infected buffaloes. CONCLUSIONS: Frequent occurrence of babesiosis among apparently healthy bovines in Egypt, suggests the need for appropriately designed prevalence studies in this country. Infected bovine, but not buffalo, populations often present hematological disorders compatible with intravascular hemolysis and thrombocytopenia.