CCL24 regulates biliary inflammation and fibrosis in primary sclerosing cholangitis

ˆCCL24 is a pro-fibrotic, pro-inflammatory chemokine expressed in several chronic fibrotic diseases. In the liver, CCL24 plays a role in fibrosis and inflammation, and blocking CCL24 led to reduced liver injury in experimental models. We studied the role of CCL24 in primary sclerosing cholangitis (PSC) and evaluated the potential therapeutic effect of blocking CCL24 in this disease. Multidrug resistance gene 2-knockout (Mdr2-/-) mice demonstrated CCL24 expression in liver macrophages and were used as a relevant experimental PSC model. CCL24-neutralizing monoclonal antibody, CM-101, significantly improved inflammation, fibrosis, and cholestasis-related markers in the biliary area. Moreover, using spatial transcriptomics, we observed reduced proliferation and senescence of cholangiocytes following CCL24 neutralization. Next, we demonstrated that CCL24 expression was elevated under pro-fibrotic conditions in primary human cholangiocytes and macrophages, and it induced proliferation of primary human hepatic stellate cells and cholangiocytes, which was attenuated following CCL24 inhibition. Correspondingly, CCL24 was found to be highly expressed in liver biopsies of patients with PSC. CCL24 serum levels correlated with Enhanced Liver Fibrosis score, most notably in patients with high alkaline phosphatase levels. These results suggest that blocking CCL24 may have a therapeutic effect in patients with PSC by reducing liver inflammation, fibrosis, and cholestasis

دراسة تحليلية لاستخدام المنتجات الجيوديزية الدقيقة على دقة شبكات الاستناد الجيوديزية لمراقبة الانزياحات المقيّسة بنظم الملاحة العالمية GNSS

ترتبط المعالجة الدقيقة لشبكات مراقبة الانزياحات التي تستخدم نظم الملاحة العالمية بالأقمار الصناعية (GNSS) بشروط القياس الصارمة أولاً، وعلى مقيدات المعالجة اللاحقة ثانياً، تلك المعالجة التي تحتاج إلى توافر المنتجات الجيوديزية الدقيقة وأهمها تصحيحات المدارات التي تقدمها الخدمة الدولية لنظم الملاحة (IGS)، سواء تم استخدمها كتصحيحات للرسالة الملاحية القياسية أو كبديل كلي عنها. يهدف البحث إلى دراسة أثر استخدام منتجات المدارات الدقيقة من (IGS) على دقة الموقع ضمن تحليل شبكات مراقبة الانزياحات، ضمن معيارين، الأول يستخدم ملفات المدارات الدقيقة (*.sp3)، والثاني يدمج معها ملفات الرسالة الملاحية القياسية (*.yyp)؛ ضمن مستويين: الأول يعالج المحطات المقاسة محلياً، والثاني يعالج تلك المحطات المحلية بدعم من محطات (IGS) المحيطة بمنطقة الدراسة. أظهرت نتائج تحليل شبكة الاستناد لمراقبة انزياحات كلية الهندسة المدنية، المقاسة بالطريقة الساكنة التقليدية لنظم (GNSS)، أن دقة الشبكة التي تتكون من المحطات المحلية، باعتماد دمج الرسالة الملاحية الدقيقة من (IGS) مع الرسالة الملاحية القياسية قد بلغت (±3 mm) أفقياً و(±5 mm) شاقولياً، بمقابل (±5 mm) أفقياً و(±9 mm) شاقولياً عند استخدام الرسالة الملاحية الدقيقة من (IGS)، أما عند دعم المعالجة بمحطات (IGS) فقد بلغت الدقة بمعيار دمج (*.yyp) مع (*.sp3) (±3 mm) أفقياً و(±5 mm) شاقولياً، وهذا يؤكد ضرورة وجود الرسالة الملاحية (*.yyp) مع (*.sp3) كشرط لازم لتحسين الدقة ضمن الشبكات الجيوديزية

The Immune Interplay between Thyroid Papillary Carcinoma and Hepatic Fibrosis.

A high prevalence of thyroid papillary cancer was reported in hepatitis-C-virus (HCV) positive patients. However, the mechanistic role of hepatic-fibrosis in thyroid malignancy progressions is still unclear.We aimed to study the immune-modulatory interactions between thyroid papillary carcinoma and hepatic-fibrosis.Hepatic-fibrosis was induced in nude-nu-male mice by intra-peritoneal administration of carbon-tetrachloride. To induce thyroid-tumor, a thyroid papillary carcinoma cell line (NPA) was injected subcutaneously in the backs. Fibrotic profile was estimated by α-smooth-muscle-actin (αSMA) expression in liver tissue extracts using western-blots and RT-PCR. Intra-hepatic NK cells were isolated and stained for NK activity (CD107a) by flow cytometry. Liver histopathology (H&E staining), thyroid tumor mass and serum alanine aminotransferase (ALT), serum vascular endothelial growth factor (VEGF) and free-T4 levels were also assessed.Ex-vivo: NPA cells were co-cultured with intra-hepatic NK cells isolated from fibrotic mice with/without the tumor were analyzed for CFSE-proliferations. Both tumor groups (with/without hepatic-fibrosis) excreted higher serum free T4 levels. Hepatic-fibrosis increased tumor weight and size and serum free-T4 levels. In addition, tumor induction increased liver injury (both hepatic-fibrosis, necro-inflammation and serum ALT levels). In addition, tumor-bearing animals with hepatic-fibrosis had increased NK activity. NPA tumor-bearing animals increased fibrosis in spite of increased NK activity; probably due to a direct effect through increased serum free-T4 excretions. Serum VEGF levels were significantly increased in the fibrotic- bearing tumor groups compared to the non-fibrotic groups. In-vitro, NK cells from fibrotic tumor-bearing animals reduced proliferation of NPA cells. This decrease is attributed to increase NK cells activity in the fibrotic animals with the NPA tumors.Our results propose that NK cells although were stimulated in advanced fibrosis with tumor, they lost their anti-tumor and anti-fibrotic activity probably due to secretions of T4 and VEFG and may explain increased risk of thyroid tumors in chronic HCV patients

NPA Tumor inductions increase the severity of CCl4 related hepatic injury.

<p>CCl4-hepatic injury was evaluated by hematoxylin and eosin (H&E) staining of necro-inflammatory liver lesions and ALT serum levels. Immunohistochemical staining with H&E (5X magnification) for the four major animal groups showed necro-inflammatory lesions and cell infiltrations that were increased in the fibrotic mice receiving the NPA-tumor cells (D) as compared to fibrotic alone (C). Arrows indicate the area with lymphocyte infiltrations. No inflammatory infiltrates were seen in H&E staining of (A) naïve WT and (B) naïve mice receiving the NPA-tumor cells. (E) Serum ALT levels were in line with histological findings and showed increase from (60±25/L) in fibrotic animals without tumor to (85.5 ± 20.5 U/L) in animals with tumor and hepatic fibrosis; p-value = 0.021.</p

VEGF serum levels.

<p>Quantikine Mouse VEGF immunoassay serum levels was significantly increased in the fibrotic group bearing tumor as compared to tumor alone (p = 0.01) or fibrosis alone groups (p = 0.04). No statistical significant differences were found in VEGF serum levels between the fibrotic and non-fibrotic group with tumor induction.</p

In vitro co-culture of lymphocytes with NPA cell line.

<p>Adhered-NPA cells post co-culture with NK cells from different animal groups were analyzed for proliferation by CFSE using flow cytometry. (A) Direct co-culture of NPA cells with spleen NKs from fibrotic mice with tumor significantly decreased NPA tumor cell proliferation compared to the fibrotic mice without tumor, indicating highly stimulated NK cells effects; p-value = 0.001. (B) A representative histogram of the NPA cells following incubations with NK cells of fibrosis and tumor mice. The histogram shows CSFE-proliferations changes in day 3 and day 5 as compared to day 0 of CSFE staining. Proliferations fold changes were calculated by divided day 0 to day 5.</p

Effect of exogenous methyl jasmonate on growth, gas exchange and chlorophyll contents of soybean subjected to drought

Drought is considered as one of the major constraints to crop production worldwide. Methyl jasmonate (MJ) is a plant-signaling molecule that elicits a wide variety of plant responses ranging from morphological to molecular level. A pot-culture study was undertaken to investigate the possible role of MJ-treatment on growth, gas exchange and chlorophyll contents in soybean (Glycine max L. Merrill.) plants subjected to water stress. The soybean plants were grown under normal water supply conditions till blooming and were then subjected to moisture stress by withholding water followed by foliar application of MJ at the rate of 50 mu M. Drought stress severely hampered the growth, leaf gas-exchange attributes as well as the photosynthetic pigment contents. It was evident from the experimental results that, MJ-treatment led to further impairment in growth by inhibiting the leaf gas exchange attributes and chlorophyll contents. It is worth noted that, MJ-treatment also hampered the performance of soybean crop under well-watered conditions. In all, MJ-treatment appeared to arrest the growth, impaired leaf gas-exchange attributes and caused the loss of chlorophyll contents of soybean plants under water deficit conditions

Hepatic fibrosis increase NPA tumor weight and size.

<p>In vivo S.C injection of NPA cells model was performed as described in M&M. In this model; S.C NPA-cells tumor was induced in fibrotic and naïve (no fibrosis) animals. S.C tumors were explanted at the end of 6 weeks post S.C injection, and evaluated for tumor weight and volume. (A) and (B) show the external appearance of the tumor in the animal’s back. Tumor weight (C) and volume (D) was increased significantly from (0.13±0.06gr) and (0.28±0.18ml) in the fibrotic group to (0.05±0.025 gr) and (0.09±0.01ml) in the non-fibrotic group; p-value = 0.02 and 0.04, respectively.</p