TROP2 Expressed in the Trunk of the Ureteric Duct Regulates Branching Morphogenesis during Kidney Development

TROP2, a cell surface protein structurally related to EpCAM, is expressed in various carcinomas, though its function remains largely unknown. We examined the expression of TROP2 and EpCAM in fetal mouse tissues, and found distinct patterns in the ureteric bud of the fetal kidney, which forms a tree-like structure. The tip cells in the ureteric bud proliferate to form branches, whereas the trunk cells differentiate to form a polarized ductal structure. EpCAM was expressed throughout the ureteric bud, whereas TROP2 expression was strongest at the trunk but diminished towards the tips, indicating the distinct cell populations in the ureteric bud. The cells highly expressing TROP2 (TROP2high) were negative for Ki67, a proliferating cell marker, and TROP2 and collagen-I were co-localized to the basal membrane of the trunk cells. TROP2high cells isolated from the fetal kidney failed to attach and spread on collagen-coated plates. Using MDCK cells, a well-established model for studying the branching morphogenesis of the ureteric bud, TROP2 was shown to inhibit cell spreading and motility on collagen-coated plates, and also branching in collagen-gel cultures, which mimic the ureteric bud's microenvironment. These results together suggest that TROP2 modulates the interaction between the cells and matrix and regulates the formation of the ureteric duct by suppressing branching from the trunk during kidney development

تاثيرات الجوسيبول و ابوجوسيبول و جوسيبولون علي خصوبة ذكور الفئران وحركة الحيوانات المنوية خارج الجسم

The effect of the naturally occurring polyphenolic pigment, gossypol, and some of its metabolites, apogossypol and gossypolone, on the fertility of male rats has been studied in vivo. The normal growth of the rats was significantly retarded in gossypol and gossypolone treated groups compared to either the controls or to apogossypol group. The in vivo effect of these compounds on sperm mobility was also examined. Gossypol acetic acid and gossypolone injection (5 mg/Kg body weight) decreased significantly the sperm count and sperm motility. Whereas apogossypol (5 mg/Kg body weight) showed insignificant effects on the sperm count and sperm motility. Gossypol and gossypolone decrease significantly the sperm mobility in vitro at different concentrations studied (5 up to 40 mg/ml saline). Apogossypol, only at high concentration (40 mg/ml) indicate inhibitory effect on sperm motility. Gossypol showed intermediate effect between the highly toxic compound, gossypolone, and the less toxic one, apogossypol.تمت دراسة التأثيرات - داخل الجسم الحي - التي يحدثها الجوسيبول - وهي صبغة عديدة الفينولات من النواتج الطبيعية - ومشتقاتها ابوجوسيبول وجوسيبولون على الخصوبة في ذكور الفئران ، كما درس أيضاً تأثير المركبات الثلاثة على حركة الحيوانات المنوية خارج الجسم الحي . تأخر النمو الطبيعي للفئران المعالجة بواسطة جوسيبول وجوسيبولون بفروق احصائية واضحة مقارنة بالفئران المعالجة بواسطة ابوجوسيبول أو بحيوانات المجموعة الضابطة . وقد أدى حقن الفئران بالجوسيبولون - 5ملليجرام / كيلو جرام من وزن الجسم - إلي حدوث نقص احصائي واضح لكل من عدد و حركة الحيوانات المنوية . في حين لم يظهر حقن ابوجوسيبول أي تأثير على الخصوية ، إلا أنه أظهر تأثيرا على حركة الحيوانات المنوية خارج الجسم الحي فقط عند تركيز 40 ملليجرام / ملليلتر مذيب . ويمكن اعتبار ابوجوسيبول مانع للخصوبة فقط في التركيزات المرتفعة ، وقد أظهر الجوسيبول تأثيرا متوسطاً بين الجوسيبولون - المركب عالي السمية - و ابوجوسيبول - الأقل في درجة السمية

تأثير جوسيبول وأبوجوسيبول وجوسيبولون على نمط الأحماض الدهنية لأقسام الليبيدات في مصل الدم والسائل المنوي للفأر

he fatty acids pattern of the total lipid fractions of male rat serum and seminal fluids were studied under the effect of gossypol and some of its metabolites (Apogossypol and gossypolone). The applied dose of gossypol and its derivatives exhibits some metabolic effects such as significant decrease of the saturated fatty acids and a significant increase of polyunsaturated fatty acids. In the serum, eicosapentaenoic acid (C20:5) in sterylester fraction was significantly increased under the effect of apogossypol. While in the free fatty acids fraction, palmitic acid (C16:0) content was significantly decreased, meanwhile significant increase of polyunsaturated fatty acid (C22:5) with apogossypol and gossypolone treatment. Stearic acid (C18:0) was significantly decreased in the total phospholipids fraction due to the treatment with gossypol acetic acid, apogossypol and gossypolone. In the seminal fluid, also eicosapentaenoic acid (C20:5), in sterylester fraction, was significantly increased with apogossypol treatment. In triglycerides, free fatty acids and phospholipids fractions, mono-unsaturated fatty acids were significantly decreased after gossypol and gossypolone treatment. Also, linoleic acid was significantly increased in the main fractions (triglycerides) under the effect of gossypol and gossypolone. Gossypol and gossypolone which induced sterility, may have significant effect on NAD and NADP-dependent enzymes, ATPase and/or have local effect on the germinal cells. Therefore, apogossypol which did not cause sterility, has no effect on the process of spermatogenesis as well as the content of seminal monounsaturated fatty acids.دُرست أنماط الأحماض الدهنية لأقسام الليبيدات الكلية في مصل الدم وسائل منوي من ذكر الفأر وذلك تحت تأثير جوسيبول وبعض مشتقات الأيض الخاصة به (أبوجوسيبول وجوسيبولون ) . أظهرت النتائج أن الجرعة المستخدمة من جوسيبول وأبوجوسيبول وجوسيبولون تؤثر على أيض بعض مركبات مثل خفض الأحماض الدهنية المشبعة وزيادة معنوية للأحماض الدهنية عديدة الروابط الغير مشبعة . في مصل الدم زاد معنوياً حمض (ك 20 :5 ) في قسم استرالاستيرولات وذلك تحت تأثير أبوجوسيبول . وبينما نقص معنوياً حمض البالميتيك من قسم الأحماض الدهنية الحرة وظهر في نفس الوقت زيادة معنوية للحمض الدهني خماسي الروابط الغير مشبعة (ك 22 :5 ) بالمعالجة بأبوجوسيبول وجوسيبولون . وفي قسم الفسفوليبيدات الكلية انخفض معنوياً حمض إستياريك (ك 18 : 0 )نتكليجة للمعالجة بواسطة خلات الجوسيبول وجوسيبولون وأبوجوسيبول . في السائل المنوي زاد أيضاً معنوياً محتوى استرات ستيرولات من حمض (ك 20 :5 ) بالمعالجة بأبوجوسيبول . وفي أقسام الجلسريدات الثلاثية وقسم الأحماض الدهنية الحرة وأقسام الليبيدات الفسفاتية انخفض معنوياً محتواها من الأحماض الدهنية وحيدة الرابطة الغير مشبعة ، وذلك لمعالجتها بجوسيبول وجوسيبولون حيث ظهرت زيادة معنوية واضحة لحمض اللينوليك في القسم الرئيسي للدهون (جلسريدات ثلاثية) ، ويعطي هذا احتمالية أن مسببات العقم من جوسيبول وجوسيبولون لها تأثير معنوي على ن اد وإنزيمات تعتمد على ن اد الذي لا يسبب العقم ليس له تأثير على إنتاجية الحيوانات المنوية ولا بيئتها من الأحماض الدهنية (أحادية الروابط الغير مشبعة)

One-pot synthesis, computational chemical study, molecular docking, biological study, and in silico prediction ADME/pharmacokinetics properties of 5-substituted 1H-tetrazole derivatives

Abstract An efficient synthesis of 5-substituted 1H-tetrazoles was successfully achieved through one-pot multi-component condensation reactions of some aromatic aldehydes or indolin-2,3-dione with malononitrile and sodium azide using diverse reaction conditions to obtain considerable product yields. Furthermore, it has been achieved for the first time to construct desired products under neat condition. Molecular docking studies with CSNK2A1 receptor disclosed the lowest binding energy displayed by the dimethoxyphenyl derivative 4c with − 6.8687 kcal/mol. The synthesized tetrazoles were screened for their in-vitro cytotoxic activity against epidermoid cancer cell line (A431) and colon cancer line (HCT116) with respect to normal skin fibroblast cell line (BJ-1) using MTT assay, and antimicrobial activity against the bacteria: K. pneumonia, S. aureus, and the fungi: Candida albicans, as well as their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl assay. In addition, the toxicity of tetrazole derivative was assessed by determination of their approximate lethal dose fifty (LD50), calculated via an oral administration to rats, through measurement of ALT and bilirubin levels in serum. The antitumor results can suggest that the potent tetrazole derivative namely, 3-(3,4-dimethoxyphenyl)-2-(1H-tetrazol-5-yl)acrylonitrile (4c) could be a potential drug against epidermoid carcinoma. The antioxidant results indicated to tetrazoles exhibited great antioxidant properties even at very low doses. A molecular dynamics simulation was performed for the synthesized compounds (ligands) to investigate their tendency for binding with the active sites of protein

تأثير جوسيبول وجوسيبولون وأبوجوسيبول على الليبيدات في مصل الدم والسائل المنوي في ذكور الفئران

The neutral and polar lipids of serum and seminal fluid of male rats were investigated under the effect of gossypol acetic acid, apogossypol and gossypolone. The total phospholipids and the neutral serum lipids showed positive and negative significant correlation with the duration of gossypol acetic acid treatment, respectively. At the same time, free fatty acids content significantly increased with apogossypol and gossypol acetic acid. In seminal fluids, the total lipid fraction showed insignificant variation in spite of the correlation between triglycerides and sterylester content. In control animals, triglycerides and sterylester contents showed positive and negative correlation, respectively. Only sterylester showed a negative significant correlation with the duration of apogossypol treatment. This result may be related to the negative effect of apogossypol on fertility. The effect of gossypol acetic acid, apogossypol and gossypolone on the total phospholipids constituents of male rat serum and seminal fluid were studied. Phosphatidyl inositol was significantly decreased under the effect of gossypol acetic acid and gossypolone in the serum, and negatively significant correlated with the duration of gossypol acetic acid and gossypolone treatment in the seminal fluid. The results indicate that infertility dose of gossypol and gossypolone exhibit some metabolic effect by elevation of lipolysis and perhaps depression of phospatidyl inositol biosynthesis.تمت دراسة تأثيرات خلات الجوسيبول وأبوجوسيبول وجوسيبولون على الدهون المتعادلة والدهون القطبية في مصل السم وسائل المني في ذ كور الفئران . أظهرت مجموعة الفسفوليبيدات ومجموع الليبيدإِت المتعادلة في المصل علاقة تزايدية معنوية ويقابلها علاقة تناقصية معنوية على الترتيب مع زمن المعالجة بخلات الجوسيبول في نفس الوضآ زاد معثوياً محتوى الأحماخى الدهنية الحرة تحت تاثير الجوسيبول والأبوجوسيبول . في السائل المنل ي - أظهر مجموع الليبيدات الكلية تغي!راً غير معنوي بالنخكم من وجود علاقة معنوية بين محتواها من الجلسريدات الثلاثية سأسترات الأستينيل . في المجموعة القياسية - أمكن تحديد علاقة تزايدية معنوية بين كمية الجلسريدات الثلاثية ورْمن الدراسة يقابلها علاقة تناقصية في كمية أسترات ستيريل لنفس المجموعة . فقط تحت تا اثيرات أبوجوسيبولى أظهر علامة تثاقصية بين أسترات ستيريل وزمن المعالجة . هذه النتيجة يمكن ربطها بالتأثير السلبي للأبوجوسيبول على الخصوبة . بدراسة تأثير خلات الجوسيبول وأبوجوسيبول وجوسيبولوق على مكونات الفسفوليبيدات إِلكلية في مصل لمحم وسائل المثي لشكور الفئران وجد أن فوسفاتيهديل اللإنيزيتول بالمصل انخفضت كميتها بدرجة معنوية تحت تأثير خلات جوسيبول والجوسيبولون وكانت العلاقة تناقصية معثوية بسرمجة كبيرة بين كمية فوسفاتيديل الإنيزيتول في السائل الملنوي وزمن المعالجة بخلات الجوسيبول والجوسيبولت . تشير الثتائج إلى أن الجرعات المسببة للعقنم من جوسيبول وجوسيبولون تعطي بعض الث ثيرات على الأيض من رفع لمعدل تحلل الدهون وبالتالي خفض في معدل بناء فوسفاتيديل أثيروتول

Hepatocellular Carcinoma cells: activity of Amygdalin and Sorafenib in Targeting AMPK /mTOR and BCL-2 for anti-angiogenesis and apoptosis cell death

Abstract Background Sorafenib (Sor) is the only approved multikinase inhibitor indicated for the treatment of HCC. Previous studies have shown that amygdalin (Amy) possesses anticancer activities against several cancer cell lines; we suggested that these compounds might disrupt AMPK/mTOR and BCL-2. Therefore, the current study used integrated in vitro and in silico approaches to figure out Amy and Sor’s possible synergistic activity in targeting AMPK/mTOR and BCL-2 for anti-angiogenesis and apoptosis cell death in HepG2 cells. Results Notably, Amy demonstrated exceptional cytotoxic selectivity against HepG2 cells in comparison to normal WI-38 cells (IC50 = 5.21 mg/ml; 141.25 mg/ml), respectively. In contrast, WI-38 cells were far more sensitive to the toxicity of Sor. A substantial synergistic interaction between Amy and Sor was observed (CI50 = 0.56), which was connected to cell cycle arrest at the S and G2/M stages and increased apoptosis and potential necroptosis. Amy and Sor cotreatment resulted in the highest glutathione levels and induction of pro-autophagic genes AMPK, HGMB1, ATG5, Beclin 1, and LC3, suppressed the mTOR and BCL2 anti-apoptotic gene. Finally, the docking studies proposed that Amy binds to the active site of the AMPK enzyme, thus inhibiting its activity. This inhibition of AMPK ultimately leads to inhibition of mTOR and thus induces apoptosis in the HepG2 cells. Conclusion Although more in vivo research using animal models is needed to confirm the findings, our findings contribute to the evidence supporting Amy’s potential anticancer effectiveness as an alternative therapeutic option for HCC

Expression status of circ-SMARCA5, circ-NOL10, circ-LDLRAD3, and circ-RHOT1 in patients with colorectal cancer

Abstract Colorectal cancer (CRC) poses a significant burden on both the healthcare systems as well as individuals. The high mortality rate of CRC may be attributed to its metastatic potential, heterogeneity, and delayed diagnosis. CircRNAs are an essential class of regulatory RNAs that play significant roles in cancers. This study aimed to detect the expression status of circ-SMARCA5, circ-NOL10, circ-LDLRAD3, and circ-RHOT1 in patients with CRC. This study included 50 CRC patients, 30 individuals with colorectal diseases (non-cancer), and 20 healthy volunteers. By using real-time PCR, the relative expression of circ-SMARCA5, circ-NOL10, circ-LDLRAD3, and circ-RHOT1 was determined in the collected blood samples. In addition, ECLIA was used to quantify carcinoembryonic antigen (CEA) level. All circRNAs expression and CEA levels were significantly up-regulated in cancer patients (CRC, colon, rectum) as compared to healthy controls, except circ-SMARCA5. Moreover, there was a significant up-regulation of circRNAs in most non-cancer patients (UC, polyp, piles). Insignificant upregulation was observed in circRNAs and CEA when comparing cancer with non-cancer patients. No correlations were found between the studied parameters and most clinicopathological characteristics of cancer and non-cancer patients. Circ-SMARCA5, circ-NOL10, circ-LDLRAD3, and circ-RHOT1 were differentially expressed in patients with CRC as well as in non-cancer patients. Circ-SMARCA5 and circ-NOL10 may act as tumor suppressors, while circ-LDLRAD3 and circ-RHOT1 may be oncogenes. Circ-SMARCA5, circ-NOL10, circ-LDLRAD3, and circ-RHOT1 could be promising markers for the early detection of CRC