Association of MALAT1 and PVT1 Variants, Expression Profiles and Target miRNA-101 and miRNA-186 with Colorectal Cancer: Correlation with Epithelial-Mesenchymal Transition.

The influence of PVT1 and MALAT1 variants on colorectal cancer (CRC) susceptibility and their impact on PVT1/miRNA-186/epithelial-mesenchymal transition (EMT) and MALAT1/miRNA-101/EMT axes in CRC are unknown. We investigated the influence of PVT1 rs13255292 and MALAT1 rs3200401 on the risk of CRC and adenomatous polyps (AP), their impact on the long noncoding RNAs PVT1 and MALAT1 expression and their target miRNA-186, miRNA-101/E-cadherin pathways, along with their potential as early CRC biomarkers. Overall, 280 individuals were recruited: 140 patients with CRC, 40 patients with AP, and 100 healthy volunteers. Genotyping and serum expression profiles were assessed using qPCR. The EMT biomarker, E-cadherin, was measured by ELISA. rs3200401 was associated with increased CRC risk, whereas rs13255292 was protective. Serum PVT1 and MALAT1 were upregulated in CRC and AP patients versus healthy controls, whereas, miRNA-186, miRNA-101 and E-cadherin were downregulated in CRC versus non-CRC groups. MALAT1 showed superior diagnostic potential for CRC and predicted CRC risk among non-CRC groups in the multivariate logistic analysis. PVT1, MALAT1, miRNA-186 and miRNA-101 levels were correlated with E-cadherin, tumor stage, lymph node and distant metastasis. E-cadherin was lost in metastatic vs. non-metastatic CRC. rs3200401CC genotype carriers showed higher E-cadherin levels than CC + CT carriers. rs3200401 was correlated with lymph node status. For the first time, rs13255292 and rs3200401 are potential genetic CRC predisposition markers, with rs3200401 possibly impacting the EMT process. Serum PVT1, MALAT1, miRNA-186 and miRNA-101 are novel non-invasive diagnostic biomarkers that could improve the clinical outcome of CRC.

Impact of betanin against paracetamol and diclofenac induced hepato-renal damage in rats.

Context: Paracetamol (PAR) and diclofenac (DF) are the most popular consumed analgesics and anti-inflammatory medications.Objective: This study aimed to explore the protective effect of betanin (Bet) against PAR or DF induced hepato-renal damage in rats.Methods: Rats were randomly divided into five groups: Normal control (NC) group rats were given saline only. PAR group rats received PAR (400 mg/kg). PAR/Bet treated group rats administered PAR (400 mg/kg) plus Bet (25 mg/kg). DF group rats received DF (10 mg/kg). DF/Bet treated group rats administered DF (10 mg/kg) plus Bet (25 mg/kg). All drugs were given by gavage for 28 consecutive days.Results: PAR and DF administration in high dose and long-time induced liver and kidney injury, disrupted serum lipid profile, enhanced serum levels of inflammatory and oxidative stress markers, triggered DNA fragmentation and caused drastic changes in the histopathological pictures of the two organs. Bet supplementation succeeded to ameliorate most of the biochemical changes and protected DNA from damage as obtained from comet assay. Histological features in H&E taken to different groups also mirrors this findings.Conclusion: Bet exerted a potential anti-inflammatory and antioxidant effect against hepato-renal damage induced by PAR or DF overconsumption.

Association between endoglin/transforming growth factor beta receptors 1, 2 gene polymorphisms and the level of soluble endoglin with preeclampsia in Egyptian women.

INTRODUCTION: Preeclampsia (PE) is a pregnancy-specific hypertensive disease whose etiopathogenesis remains unclear. OBJECTIVES: This study was designed to assess association between PE and 3 single nucleotide polymorphisms (SNPs): ENG(G/A) rs11792480, TGFßR1(A/C) rs10739778 and TGFßR2(G/A) rs6550005, beside circulating soluble endoglin (sENG), oxidative stress biomarkers and nitric oxide (NO) in Egyptian women. METHODS: The study included 75 preeclamptic women stratified into 4 clinical subgroups and 50 normotensive pregnant women. Genotyping was performed by real time polymerase chain reaction-Taqman allelic discrimination. RESULTS: Preeclamptic women showed significantly increased sENG and malondialdehyde (MDA), decreased total antioxidant capacity (TAC), endothelial nitric oxide synthase (eNOS) and NO, without change in transforming growth factor beta 1 (TGFß1) versus controls. Moreover, sENG was significantly higher in severe and early than mild and late PE. Higher MDA and lower TAC and NO were observed in severe than mild PE. ENG(G/A) and TGFßR2(G/A) showed no association with PE. However, CC genotype of TGFßR1(A/C) was more frequent in controls than either PE, early-onset or severe revealing a reduced PE risk in CC genotype versus AA or AA + AC. Importantly, patients carrying AA genotype had higher SBP and MDA with lower TAC, gestational age at delivery (GA) and birth weight than those carrying CC genotype. CONCLUSIONS: Excessive sENG release with decreased eNOS/NO may be involved in PE pathogenesis. Women who carry C allele or CC genotype of TGFßR1(A/C) may be less prone to develop PE.

Protective effects of betanin against paracetamol and diclofenac induced neurotoxicity and endocrine disruption in rats.

Context: Overconsumption of paracetamol (PAR) and diclofenac (DF) have been reported to induce neurotoxicity and endocrine disruption. Objective: The current study was designed to explore the protective potential of betanin against PAR and DF inducing neurotoxicity and endocrine disruption in a rat model. Material and Methods: Forty rats were equally divided into five groups: group I served as control, group II received PAR (400 mg/kg), group III received PAR plus betanin (25 mg/kg), group IV received DF (10 mg/kg) and group V received DF plus betanin orally for 28 consecutive days. Thyroid axis hormones, sex hormone, neurotransmitters, paraoxonase-1, hemeoxygenase-1 and nuclear factor-2 were measured by ELISA. While, the oxidative stress markers were colorimetrically estimated. Moreover, DNA damage and histopathological picture of the brains were investigated. Results: A marked reduction in thyroid axis hormones, brain neurotransmitters and serum testosterone as well as enhanced oxidative stress and brain DNA damage accompanied by drastic changes in the brain histopathological picture were recorded in the challenged PAR and DF groups. Betanin supplementation ameliorated most of the biochemical and histopathological changes induced by PAR or DF. Conclusion: The study suggests betanin of potential protective effects against neurotoxicity and endocrine disruption induced by PAR and DF overconsumption.

Evaluation and screening of mRNA S100A genes as serological biomarkers in different stages of bladder cancer in Egypt.

Calcium-binding proteins S100A are multifunctional proteins that show altered expression in various diseases and cancers. This study aimed at validating an easier and less time-consuming technique to evaluate the value of combined use of messenger RNA (mRNA) S100A genes in comparison and combination with voided urine cytology in detection of bladder cancer patients. Blood and urine specimens were collected from patients (n = 120) with histologically confirmed bladder carcinoma who are classified according to bladder cancer stage into four groups and from healthy volunteers (n = 30). Histopathology examination, bilharzias antibody detection, urine cytology, and mRNA expression of S100A genes were estimated for all subjects by real time polymerase chain reaction (RT-PCR). Results indicate that each of the investigated S100A genes can be used as diagnostic marker for bladder cancer. Both S100A4 and S100A6 can be used to differentiate between different stages of bladder cancer. S100A7 can be used for the diagnosis of squamous cell carcinoma. Both S100A8 and S100A9 can be used for detection of invasive bladder carcinoma while S100A11 can be used for early detection of superficial bladder carcinoma. The overall sensitivity and specificity for the studied S100A genes ranged from 73 to 90 and 84 to 92, respectively. The combined use of urine cytology with the investigated S100A genes increased sensitivity from 56 % up to a range of 87-96 %. In conclusion, serum S100A genes can be useful as potential serological biomarkers for bladder cancer, and combined use of urine cytology with S100A genes can improve the sensitivity for detection of bladder cancer.

The lncRNAs UCA1 and CRNDE target miR-145/TLR4/NF-қB/TNF-α axis in acetic acid-induced ulcerative colitis model: The beneficial role of 3,3-Diindolylmethane.

INTRODUCTION: Ulcerative colitis (UC) is a chronic disease that alters the colonic and rectal mucosa. The high prevalence rates of UC make it a worldwide healthcare problem. However, its underlying molecular mechanisms remain vague. AIM OF THE STUDY: To investigate the molecular mechanisms underlying UC and to study the cross-talk among the regulatory role of the lncRNAs UCA1, CRNDE, and miR-145 on TLR4/NF-κB/TNF-α signaling pathway. Moreover, the study was extended to examine the beneficial effects of 3,3-Diindolylmethane (DIM) on relieving UC. METHODS: UC was induced in rats by injecting 2 ml of 4% acetic acid (AA) solution transrectally. After 24 h, rats were treated with either DIM (20 mg/kg) or sulphasalazine (SSZ) (500 mg/kg) orally for 7 days. RESULTS: The present study revealed that the gene expression of the lncRNAs UCA1 and CRNDE were significantly upregulated in the AA-induced UC model compared with the control group, whereas miR-145 was significantly downregulated. There was a significant association between the expression of these non-coding RNAs and TLR4/ NF-κB/TNF-α axis as well as malondialdehyde and glutathione levels. Favorably, the DIM-treated group showed significant downregulation of the lncRNAs UCA1 and CRNDE along with upregulated miR-145 compared with the AA-induced UC model. Furthermore, DIM showed remarkable inhibition of the TLR4/ NF-κB /TNF-α cascade compared with non-treated UC rats. CONCLUSIONS: The present study is the first to document the interrelated role of the lncRNAs UCA1 and CRNDE in UC via orchestrating miR-145/TLR4/ NF-κB /TNF-α inflammatory cascade. Furthermore, the study demonstrated a new molecular basis for the pleiotropic activities of DIM in relieving UC.

MAGI2-AS3 and miR-374b-5p as Putative Regulators of Multiple Sclerosis via Modulating the PTEN/AKT/IRF-3/IFN-ß Axis: New Clinical Insights.

Multiple sclerosis (MS) is a chronic disease and one of the leading causes of disability in young adults. The current study aims to investigate the pathogenesis of MS via studying the regulatory role of novel lncRNA MAGI2-AS3 in miR-374b-5p and their downstream targets PTEN/AKT/IRF-3/IFN-ß and the relationship of this pathway with disease severity. Moreover, it aims to assess the role of MAGI2-AS3/miR-374b-5p as diagnostic and/or prognostic biomarkers for MS. Overall, 150 contributors were recruited: 100 patients with MS and 50 healthy volunteers. Gene expression of MAGI2-AS3, miR-374b-5p, PTEN, AKT, and IRF-3 were assessed using RT-qPCR, and IFN-ß was measured by ELISA. Compared with the healthy control group, serum MAGI2-AS3 and PTEN were downregulated in MS patients, whereas miR-374b-5p, PI3K, AKT, IRF-3, and IFN-ß were upregulated in MS patients. Furthermore, MAGI2-AS3 was downregulated, while miR-374b-5p was upregulated in MS patients with an expanded disability status scale (EDSS) ≥3.5, compared to patients with an EDSS <3.5. Receiver-operating-characteristic curve analysis revealed that MAGI2-AS3 and miR-374b-5p can be used in the diagnosis of MS. Remarkably, multivariate logistic analysis revealed that MAGI2-AS3, miR-374b-5p, PTEN, and AKT act as independent variables in MS. Moreover, MAGI2-AS3 was directly correlated with PTEN and inversely correlated with miR-374b-5p, AKT, and EDSS. Regarding miR-374b-5p, it was positively correlated with AKT and EDSS. In conclusion, the study showed for the first time that the crosstalk between MAGI2-AS3 and miR-374b-5p could affect the AKT/IRF3/IFN-ß axis in MS. Interestingly, MAGI2-AS3 and miR-374b-5p could be genetic noninvasive biomarkers for MS.

LncRNA NEAT1 and MALAT1 are involved in polycystic ovary syndrome pathogenesis by functioning as competing endogenous RNAs to control the expression of PCOS-related target genes.

Accumulating evidence has shown an abnormal expression of several non-coding RNAs in ovarian tissues which might be closely linked with the pathogenesis of PCOS. The aim of this study was to identify competing endogenous (ce) RNA network: long non-coding RNA (lncRNA), microRNA (miRNA) and their target genes: androgen receptor (AR), follistatin (FST) and insulin receptor substrate-2 (IRS-2), which are relevant to PCOS, to underline the molecular pathogenesis of PCOS and assist in early diagnosis and treatment. Bioinformatic analysis was performed to retrieve a ceRNA network: [lncRNA (NEAT1 and MALAT1) - miRNA (miR-30a-5p and miR-30d-5p) - mRNA (AR, FST and IRS-2)] linked to PCOS. Expression of the selected RNAs was examined by qPCR in peripheral blood leukocytes obtained from 73 PCOS patients (41 obese and 32 non-obese) and 31 healthy controls. PCOS patients showed significantly higher expression levels of NEAT1, miR-30a-5p, AR, FST and IRS-2, with significantly lower expression levels of MALAT1 and miR-30d-5p relative to controls especially in obese versus non-obese patients. Receiver operating characteristic (ROC) curve analysis indicated that most of the selected RNAs could serve as potential early diagnostic markers for PCOS with the highest efficiency obtained upon combining NEAT1 and miR-30d-5p or MALAT1 and miR-30a-5p with either of PCOS target genes. Moreover, all addressed RNAs had been proved as potential predictors of PCOS. The obtained data of ceRNA network raised the possibility that NEAT1 overexpression may increase the expression levels of AR, FST and IRS-2 by sponging miR-30d-5p, while low expression of MALAT1 may allow higher expression of the above genes via increasing miR-30a-5p, suggesting their involvement in PCOS pathogenesis and promising role for future diagnosis and targeted therapy.

Protective effect of ellagic acid and pumpkin seed oil against methotrexate-induced small intestine damage in rats.

Gastrointestinal toxicity is one of the most serious side effects in the methotrexate (MTX) treatment. This study was designed to investigate whether ellagic acid (EA) and/or pumpkin seed oil (PSO) had a protective effect on MTX-induced small intestine damage. Forty albino rats were randomized into five groups of 8 rats each. Group I served as a normal control group. In Group II, MTX was administered as a single dose (20 mg/kg) intraperitoneally. Groups III, IV and V were pre-treated respectively with either PSO (40 mg/kg), EA (10 mg/kg) or 0.2% DMSO (vehicle control) orally every day by gavage for 5 days and then they received MTX. All animals were sacrificed 5 days after the intraperitoneal injection of MTX for histopathological examination, estimation of serum prostaglandin E2 (PGE2) level, assay of tissue malondialdehyde (MDA), reduced glutathione (GSH) and nitric oxide (NO) levels and myloperoxidase (MPO), xanthine oxidase (XO) and adenosine deaminase (AD) activities. Administration of EA and/or PSO decreased the intestinal damage, PGE2, MDA and NO levels and MPO, XO and AD activities and increased GSH level. These results suggest that EA and PSO protect the small intestine of rats from MTX-induced damage through their antioxidant and anti-inflammatory effects and thus have potential as a promising drug in the prevention of undesired side effects of MTX.

Omeprazole and Spirulina Platensis Ameliorate Steatohepatitis in Experimental Nonalcoholic Fatty Liver Disease.

Background: Nonalcoholic fatty liver disease (NAFLD) is a leading cause of liver damage, and it affects about 24% of the population worldwide. This study aimed to investigate the effect of omeprazole and Spirulina platensis on hepatic and serum biochemical alterations in NAFLD induced by high-fat diet (HFD). Methods: Male Wistar rats were divided into four groups; one served as a normal control. The other groups received HFD and subdivided into three subgroups; one was left untreated and the other two groups were treated orally with either omeprazole (10 mg/kg) or Spirulina (1000 mg/kg) for 30 consecutive days. Results: Omeprazole successfully decreased elevated serum pentraxin-3 (PTX-3) and cytokeratin-18 (CK-18) levels and hepatic sterol regulatory element binding protein-1c (SREBP-1c) expression, while Spirulina had better impact on decreasing liver function enzymes, lipid profile, and nuclear factor erythroid 2-related factor 2 (Nrf-2) levels compared to omeprazole. Both treatments had similar effects on normalizing glucose homeostasis, decreasing insulin resistance, and improving adipocytokine levels. Conclusion: Effects of omeprazole and Spirulina on markers of NAFLD appeared to be mediated by regulation of inflammatory, apoptotic, and oxidative mediators. Thus, omeprazole and Spirulina may find use as promising adjuvant therapy to ameliorate NAFLD. Research ethical committee of the faculty of pharmacy, Cairo University approved the research (BC 1479).

Bidirectional Association Between Psoriasis and Obesity: Benefits and Risks.

Psoriasis is a chronic, immune-mediated inflammatory skin disease that is associated with several comorbidities such as obesity. This study was designed to estimate the possibility of utilizing psoriasin, nestin, keratin-16 (Krt16), and interleukin-21 (IL-21) as biochemical markers of psoriasis, to correlate these candidate psoriatic markers with biomarkers of obesity [body mass index (BMI), leptin, and resistin], and to elucidate the bidirectional association between obesity and psoriasis. Blood samples were collected from all participants (n = 108) who were classified according to their BMI into 4 groups: healthy control, obese, psoriatic, and obese psoriatic group. Plasma psoriasin, nestin, Krt16, IL-21, leptin, and resistin were estimated for all subjects. Psoriasin, nestin, Krt16, IL-21, leptin, and resistin were significantly elevated in psoriatic and obese psoriatic groups. However, only leptin, resistin, IL-21, and Krt16 were significantly increased in the obese group compared with the control group. Leptin and resistin showed significant positive correlations with psoriasis area and severity index score, psoriasin, nestin, Krt16, and IL-21. Cutoff values for psoriasin, nestin, Krt16, and IL-21 were 187.5 ng/mL, 1825 pg/mL, 33.1 ng/mL, and 128.6 ng/L, respectively. In conclusion, psoriasin, nestin, Krt16, and IL-21 can be utilized as biochemical markers of psoriasis; these psoriatic markers are significantly positively correlated with obesity biomarkers, and obesity can be considered a risk factor and/or consequence of psoriasis.

Resveratrol and Montelukast Alleviate Paraquat-Induced Hepatic Injury in Mice: Modulation of Oxidative Stress, Inflammation, and Apoptosis.

Paraquat (PQ) is one of the most used herbicide worldwide. Its cytotoxicity is attributed to reactive radical generation. Resveratrol (Res) and montelukast (MK) have anti-inflammatory and antioxidant properties. The protective effects of Res, MK, or their combination against PQ-induced acute liver injury have not been investigated before. Therefore, we explored the protective potential of Res and/or MK against PQ hepatic toxicity in a mouse model. Mice were randomly assigned to five groups: group I served as the normal control and group II received a single dose of PQ (50 mg/kg, i.p.). Groups III, IV, and V received PQ plus oral Res (5 mg/kg/day), MK (10 mg/kg/day), and Res/MK combination, respectively. Res and/or MK reduced PQ-induced liver injury, evidenced by normalization of serum total protein, ALT, and AST. Res and/or MK significantly reversed PQ-induced oxidative stress markers glutathione and malondialdehyde. Res and/or MK significantly reduced PQ-induced inflammation reflected in TNF-α levels. Furthermore, Res and/or MK reversed PQ-induced apoptosis assessed by differential expression of p53, Bax, and Bcl-2. Histopathologic examination supported the biochemical findings. Although Res and MK displayed antioxidative, anti-inflammatory, and antiapoptotic activities, their combination was not always synergistic.