Pluripotent stem cells isolated from umbilical cord form embryonic like bodies in a mesenchymal layer culture.

Recently the matrix of umbilical cord began to use as an alternative source of stem cells additionally to the blood of umbilical cord. Umbilical cord has been used mainly for mesenchymal stem cell banking. The immunological characteristics of mesenchymal stem cells in combination with their ability to avoid rejection make them an attractive biological material for transplantations. In this study the isolation of small in size pluripotent stem cells from umbilical cord expressing early transcription factors with characteristics that resemble to embryonic stem cells is investigated. Pluripotent stem cells were isolated from human umbilical cords, by a new strategy method based on unique characteristics such as the small size and the positivity on early transcription factors OCT and Nanog. An enriched population of CXCR4(+) OCT(+) Nanog(+) CD45(-) small stem cells from the cord was isolated. This fraction was able to create alkaline phosphatase positive like spheres forms in a mesenchymal layer with multilineage differentiation capacity. Our results were assessed by RT PCR and electophoresis for the pluripotent genes. These data suggest that umbilical cord provides an attractive source not only of mesenchymal stem cells but moreover of pluripotent stem cells. The method described herein should be applied in the field of stem cell banking in addition to the classical umbilical cord harvesting method. Isolation of a population of cells with pluripotent characteristics from umbilical cord. Adoption of a second centrifugation step for the pluripotent stem isolation. Increasing the value of the cord and explaining the pluripotency. This work will enhance the value of umbilical cord harvesting.

Placenta perfusion has hematopoietic and mesenchymal progenitor stem cell potential.

BACKGROUND: Placenta is a valuable source of stem cells for cell therapy and future application in the field of regenerative medicine. This is due to the plasticity and the immunomodulatory effects of the stem cells that it contains. In this study we present a totally closed method for hematopoietic and nonhematopoietic stem cell isolation from human term placenta. STUDY DESIGN AND METHODS: Sixty-eight placenta units were collected and manipulated for the residual fetal blood drainage. After delivery, placenta flushing with citrate-phosphate-dextrose-adenine was evaluated. RESULTS: Placenta flushing using a totally closed system led to a significant amount of hematopoietic progenitor cells and multipotent mesenchymal stem cells (MSCs) without additional microbial risk, free of maternal cell contamination. CONCLUSION: Traditionally discarded after childbirth, the term placenta now appears to be an easily accessible and abundant source of diverse origin stem cells suitable for banking strategies and for future clinical applications, including adult therapy.

Oral calcium supplementation ambulatory blood pressure and relation to changes in intracellular ions and sodium-hydrogen exchange.

BACKGROUND: Calcium (Ca2+) supplementation has been shown paradoxically to reduce intracellular Ca2+ and induce vascular relaxation. The aim of the study was to assess 24-h blood pressure (BP) change after Ca2+ supplementation and to investigate its relation to changes in intracellular ions and the activity of the first isoform of sodium-hydrogen exchange (NHE-1) in subjects with hypertension and type 2 diabetes. METHODS: This parallel, randomized controlled, single-blinded trial, consisted of 31 patients with type 2 diabetes, and hypertension who were allocated to receive 1,500 mg of Ca2+ per day (n = 15) or no treatment (n = 16) for 8 weeks. RESULTS: In the Ca2+ group a decrease of 1.7 +/- 2.7 mm Hg (mean +/- SE) P = 0.52 for mean 24-h systolic BP (SBP) and 2.1 +/- 1.5 mm Hg, P = 0.19 for mean 24-h diastolic BP (DBP) was recorded. Whereas in the control group an increase of 1.4 +/- 2.7 mm Hg, P = 0.59 for mean 24-h SBP and 1.2 +/- 2.8 mm Hg, P = 0.83 for mean 24-h DBP was observed. Intraplatelet Ca2+ decreased whereas intraplatelet magnesium (Mg2+) and erythrocyte K+ increased in the intervention group. Change in mean 24-h SBP in the pooled group correlated with both change in intraplatelet Ca2+ (r = 0.49, P < 0.05) and NHE-1 activity (r = 0.6, P < 0.001). The contribution of intraplatelet Ca2+ was attenuated when both parameters were entered in a multivariate regression model. CONCLUSIONS: The present study shows a weak, statistically nonsignificant trend towards association of Ca2+ supplementation on 24-h BP in hypertensive subjects with type 2 diabetes. However, our results indicated an interrelation of [Ca2+]i levels and NHE-1 activity on BP in patients with hypertension and type 2 diabetes.

Further examination of antiradical properties of Crocus sativus stigmas extract rich in crocins.

Data for antiradical properties of saffron extract and its bioactive constituents (crocins, crocetin) are limited and poorly discussed in comparison with those of extracts containing potent scavengers. Further examination was sought using the Folin-Ciocalteu (F-C) reagent and various free radical species produced in cell-free or cell model systems. Oregano and turmeric methanol extracts, rich in well established scavengers, and also crocetin, rosmarinic acid, and curcumin, representing the major types of constituents in the three studied extracts, were used as "reference". On the same weight basis, saffron extract activity was found to be rather negligible in all cell-free systems with regard to that found for reference ones. On the contrary, in the human monocyte system, saffron extracts or free crocetin were found to reduce ROS production as effectively as the phenolic antioxidants. Our findings point out that saffron extracts exhibit a remarkable intracellular antioxidant activity that cannot be revealed using assays repeatedly applied to the evaluation of phenolic-type antioxidants.

Prooxidant-antioxidant balance, peroxide and catalase activity in the aqueous humour and serum of patients with exfoliation syndrome or exfoliative glaucoma.

BACKGROUND: Oxidative stress plays an important role in the pathobiology of exfoliation syndrome (XFS) and exfoliative glaucoma (XFG). METHODS: We investigated the prooxidant-antioxidant balance (PAB) in aqueous humour and serum samples of 20 consecutive cases of XFS, 20 of XFG, and 20 age-matched controls, employing a recently described novel assay. The activity of catalase and the levels of (hydrogen) peroxide were also measured in these samples. RESULTS: There was no significant difference between the PAB in the aqueous humour of the XFS group (82.5 +/- 10 AU) and age-matched control patients (78.9 +/- 13.4 AU; p > 0.05). A significant shift of the PAB balance in favour of oxidants was detected in the XFG group (90.2 +/- 7.6 AU) compared with controls (p < 0.001). In the serum of patients with XFS (138.8 +/- 13.2 AU) and XFG (124.08 +/- 13.50 AU), PAB was significantly altered in favour of oxidants as compared to age-matched controls (114.9 +/- 9.91 AU); p < 0.001). Catalase activity in the aqueous from XFS (10.1 +/- 4.5 U/ml) and XFG (12.2 +/- 6 U/ml) patients was significantly lower than that measured in the normal aqueous (14.6 +/- 1.9 U/ml). Similarly, a significantly lower catalase activity was found in XFS (103 +/- 21.4 U/ml) and XFG (116 +/- 38 U/ml) serum samples compared with controls (189.6 +/- 84.3 U/ml). Finally, (hydrogen) peroxide concentration in aqueous and serum samples from patients with XFS (aqueous: 26.9 +/- 6.6 microM; serum: 41 +/- 10 microM) and XFG (aqueous: 21.7 +/- 7 microM; serum: 32 +/- 4 microM) were significantly higher than that of the controls (aqueous: 9.6 +/- 5.8 microM; serum: 24 +/- 9 microM; p < 0.001). CONCLUSIONS: These findings suggest that in XFS oxidative stress is counterbalanced in the aqueous, whereas the development of XFG is accompanied by a disruption of this balance in favour of oxidants.

Serum ghrelin, leptin and adiponectin levels before and after weight loss: comparison of three methods of treatment--a prospective study.

BACKGROUND: Ghrelin is a peptide hormone with orexigenic properties, primarily produced by the stomach. Leptin and adiponectin are the two adiposity products that participate in body weight control. Leptin always decreases and adiponectin increases after weight loss. Different changes in fasting ghrelin levels have been reported following bariatric surgery. In this study, we compare the changes in fasting ghrelin, leptin and adiponectin levels in 3 groups of patients who achieved weight loss by either diet, MacLean vertical banded gastroplasty (VBG) or biliopancreatic diversion with duodenal switch (BPD-DS). METHODS: Serum fasting ghrelin, leptin and adiponectin concentration was measured in 40 obese patients who achieved weight loss by either diet (n=14), VBG (n=13) or BPD-DS (n=13), before and after weight loss. The follow-up period was 18 months for BPD-DS and VBG and 6 months for diet. Serum ghrelin level was measured by ELISA. RESULTS: BMI was significantly decreased in all 3 groups: 9.2+/-2.4% (P<0.01) following diet, 38.47+/-7.26% (P<0.01) after VBG, and 42.88+/-9.09% after BPD-DS (P<0.01). Serum fasting ghrelin level increased after diet (110.45+/-117.84%, P=0.002) and VBG (65.48+/-92.93%, P=0.001),but decreased after BPD-DS (-21.63+/-28.63%, P=0.019). Leptin concentration decreased and adiponectin increased in all groups. CONCLUSIONS: Unlike after diet or gastric restrictive surgery, BPD-DS is associated with markedly suppressed ghrelin levels, possibly contributing to the weight-reducing effect of this operation. Sleeve gastrectomy seems to be the main cause of this reduction.

Latanoprost therapy reduces the levels of TGF beta 1 and gelatinases in the aqueous humour of patients with exfoliative glaucoma.

The aim of the present study was to evaluate the effect of latanoprost monotherapy on the aqueous humour concentrations of TGF-beta1, MMP-2, TIMP-2, MMP-9 and gelatinolytic activity in patients treated for exfoliative glaucoma (XFG). Aqueous samples from 50 XFG patients treated with latanoprost and 50 age-matched XFG patients treated with timolol were collected during phacoemulsification cataract surgery. The concentrations of TGF-beta1, MMP-2, TIMP-2, MMP-9 and gelatinase activity were determined by commercial immunoassays. The mean active TGF-beta1 concentration in the aqueous was significantly lower in XFG patients treated with latanoprost compared with those treated with timolol (3.1 +/- 0.65 vs 13.4 +/- 1.5 pg ml(-1)); (P = 0.0014). The mean total MMP-2 concentration was lower in latanoprost treated patients (31.75 +/- 3.8 vs 81.5 +/- 7.2 ng ml(-1)); (P < 0.0001). The TIMP-2 concentration was also lower in XFG-latanoprost treated patients (73.8 +/- 6.81 vs 101.28 +/- 7.29 ng ml(-1)); (P = 0.0096). Latanoprost monotherapy has a marked effect on the aqueous concentration of TGF-beta1, MMP-2 and TIMP-2 in XFG patients. A better understanding of its effect on the pathobiology of the disease may lead to its earlier use in the disease process to prevent progression from XFS to XFG.

Stimulation of Na+/H+ antiport and pyruvate kinase activities by high glucose concentration in human erythrocytes.

This study aims to demonstrate the effect of high glucose concentrations on NHE-1 and PK activities and investigate the implicated signal transduction pathways. Erythrocytes drawn from healthy volunteers were incubated in the presence of 5 or 50 mM of glucose, fructose, galactose or mannitol. When appropriate, specific inhibitors of NHE-1, PKC or p42/44 MAPK were used. Erythrocyte NHE-1 activity has been estimated by fluorometrical determination of the intracellular pH and quantification of sodium uptake using 22Na. Pyruvate kinase activity was measured by a NADH-lactate dehydrogenase enzymatic assay. p42/44 MAPK activity was assessed with a specific enzyme linked immunosorbent assay (ELISA). Increased concentrations of glucose but not galactose, fructose or mannitol enhanced erythrocyte NHE-1, PK and p42/44 MAPK activity. Inhibition of PKC, counteracted these effects of glucose. Similarly, inhibition of NHE 1 abolished the effect of high glucose on PK and p42/44 MAPK as well. Finally, inhibition of p42/44 MAPK also hindered the effect of glucose on NHE-1 and PK activities. The data of the present study indicate an acute effect of glucose on signal transduction pathways in human erythrocytes. This pathway involves NHE-1, PKC, and p42/44 MAPK. A positive feedback between NHE 1 and p42/44 MAPK is suggested.

Ascorbic acid concentration is reduced in the aqueous humor of patients with exfoliation syndrome.

PURPOSE: To investigate whether there is a role for ascorbic acid in the development of exfoliation syndrome (XFS). DESIGN: A case-control study was undertaken that included consecutive patients with and without XFS in whom cataract surgery was indicated. Patients with ophthalmic conditions other than XFS and conditions that may influence ascorbic acid levels were excluded. METHODS: A prospective institutional study was undertaken. A small volume of aqueous humor was aspirated at the beginning of phacoemulsification cataract surgery. Eighty aqueous samples, 40 samples from 40 eyes of 40 cataract patients with XFS and 40 samples from 40 eyes of 40 age matched cataract patients without XFS, were collected and analyzed. Ascorbic acid concentration was evaluated in the aqueous samples with a microplate assay method. RESULTS: The mean +/- SD concentration of ascorbic acid in the aqueous from patients with XFS (0.86 +/- 0.43 mM; range, 0.12 to 1.7 mM) was significantly lower than the concentration of ascorbic acid found in the aqueous of age-matched control patients (1.15 +/- 0.50 mM; range 0.42 to 3.1 mM; P =.0068). Total mean protein concentration was found to be significantly higher in the XFS group (481.1 +/- 196.8 pg/dl versus 336.3 +/- 86.4 pg/dl in the controls; P <.0001). Nevertheless, no correlation could be established between ascorbic level and protein concentration. CONCLUSIONS: A significantly reduced mean level of ascorbic acid was observed in the aqueous humor of patients with XFS. In view of the fact that ascorbic acid is a major protective factor against free radical action, a role for free radical action is possible in the pathobiology of XFS.