Assessment of the relationship between serum soluble Klotho and carotid intima-media thickness and left ventricular dysfunction in hemodialysis patients

BACKGROUND: The aim of our study was to assess the relationship between soluble Klotho (s-Klotho) and carotid intima-media thickness (CIMT) and left ventricular (LV) dysfunction in hemodialysis (HD) patients. METHODS: This is a cross-sectional study conducted on 88 patients with end-stage renal disease on regular HD. Serum levels of calcium, phosphorus, parathyroid hormone, and C-reactive protein were measured. The serum levels of s-Klotho and fibroblast growth factor-23 (FGF-23) were measured using an Enzyme linked immunosorbent assay (ELISA) kit. Echocardiography and measurement of CIMT were also conducted. The studied patients were divided according to the median s-Klotho level into 2 groups: patients with low s-Klotho (Group I) and patients with high s-Klotho (Group II). RESULTS: Mean value of s-Klotho was significantly low in HD patients compared to controls (P = 0.001), and mean value of FGF-23 was significantly high in HD patients compared to controls (P = 0.001). The mean values of parathyroid hormone, FGF-23, and phosphorus were significantly high in Group I compared to Group II, whereas the mean value of serum calcium was significantly low in Group I compared to Group II. The mean values of CIMT, LV mass (LVM), LVM index, and LV ejection fraction (LVEF) were high in Group I compared to Group II. Patients with low s-Klotho had significantly more coronary artery disease (CAD). In a regression analysis of s-Klotho with different markers of cardiovascular diseases, s-Klotho showed significant association with CIMT, LVEF, and CAD, but not with LVM and LVM index. CONCLUSION: The present study showed that patients with a low s-Klotho were more often associated with increased CIMT, LV dysfunction, and CAD, and it seems that there was independent association between s-Klotho and CIMT, LVEF, and CAD.

In vitro differentiation of mice bone marrow cells into hepatocyte-lik cells

Recent publications showed that highly selected hone marrow derived stem cells can differentiate into hepatocyte like cells. Bone marrow, considered as a rich source of adult stem cells, contains not only hematopoietic and endothelial stem cells, but also precursors of other tissues of mesenchymal origin. The different growth factors used in the culture media are critical factors directly affecting the hepatocytic differentiation and expansion process. Differentiation of adult bone marrow stem cells [BMC] into hepatocyte-like cells is commonly performed by continuous exposure to a cytokines-cocklail. Here, we investigated whether the in vitro differentiation efficacy can be enhanced by sequential addition of liver-specific factors in a time-dependent order that closely resembles their secretion pattern during in vivo liver embryogenesis. Fibroblast growth factors [FGFa and b], Leukeamia inhibitory factor [L1F] Stem cell factor [SCF], Hepatocyte growth factor [HGF] and Oncostatin M [OSM] were used as differentiation factors. Mice bone marrow cells were cultured for one week in a primary culture media and the adherent cells were then subcultured in the directed differentiation media including the above mentioned growth factors added either as a cocktail or in a sequential manner for comparison. In the course of cell differentiation, cell morphology was observed, and the expression of albumin by the transdif Terentiated cells was examined by immunofluorescence and Western blot analysis. Some epithelial-like cells or polygonal hepatocyte like cells appeared in the directed differentiation medium within 12 days, and the number and size of colonies of epithehial-like cells or polygonal cells increased in the course of the cell directed differentiation. The sequential addition of growth factors setup featured relatively earlier and more intense differentiation to hepatocyte like cells. By immunofluoresence analysis albumin expressions appeared, lasted and increased throughout the later directed differentiation. Albumin was found in the cell membrane and scattered in the cytoplasm by immunofluorescent staining. On day 21, the ratio of albumin-positive cells was 85% in the sequentially exposed cells in contrast to the cocktail treatment where the ratio of albumin expressing cells was 65%. We propose that the addition of the hepatocyte growth factor to the culture media is the pivotal step in the trans-differentiation protocol. Moreover the sequential induction of the differentiation process, analogous to in vivo liver development, is crucial for in vitro differentiation of adult mice BMC into functional hepatocyte-like cells

Estimation of protease activity in Schistosoma mansoni life cycle stages and Biomphalaria Alexandria after cystein protease inhibitor

The efficacy of cysteine inhibitor on murine Schistosom mansoni was studied when the injection of inhibitor to infected mice for 10 successive days was either starting on day 1 [group 1] or on day 21 [group2] or on day 45 [group3] post infection. Eight weeks post infection animals were sacrificed and subjected for parasitological, histological and physiological assessments. Then levels of proteases activity were assayed in normal, infected and infected treated mice in serum, livers, intestine, as well as, in S. mansoni worms and ova collected from different groups under study. Also, B. alexandrina snails were exposed to LC5, LC10 and LC25 of each of bayluscide and A. arvensis then their hemolemph protease activity were assessed after 1 day, 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks of exposure and compared with that assessed in hemolemph of control snails. Both the parasitological and physiological studies gave rise to the same conclusion denoting that treatment with cysteine protease inhibitor, phenyl vinyl sulfone, at 45 day post infection could reduce the worm burden, egg tissue load, granuloma diameter, ova hatchability, proteases activity in S. mansoni worms and ova. This treatment at the same time helps mice to attain approximately normal physiological state as expressed by their serum proteases activity. The findings representing toxicity and physiology studies gave rise to the same conclusion, emphasizing that the chemical molluscicides seriously affects snail physiology exerting irreversible alterations, while plant molluscicides has effect under which snails can acclimatize their physiology to survive