Human serum promotes the proliferation but not the stemness genes expression of human adipose-derived stem cells

Recently human adipose-derived stem cells (ASCs) have shown much therapeutic potential in regenerative medicine. However, fetal bovine serum (FBS) used in culturing human cells may give risk to viral and prion transmission as well as immune rejection. Human serum (HS) is a safer growth supplement in human cell culture but its effects have not been well established. Therefore the objectives of this study were to compare the effects of HS versus FBS on the proliferation and stemness gene expression of ASCs. ASCs were cultured for 5 passages in medium supplemented with either 10% HS or 10% FBS. ASCs proliferation rate and viability were determined at every passage. Total RNA was extracted at passage 5 (P5) and quantitative PCR was carried out to determine the stemness gene expression level of SOX-2, Nanog3, BST-1, REX-1, ABCG2 and FGF-4. The results showed ASC cultured in 10% HS scored greater proliferation rates and viability compared to 10% FBS. ASCs proliferated significantly faster in 10% HS compared to 10% FBS at P2, P3, and P4 (p < 0.05). In quantitative gene expression analysis, ASCs cultured in 10% FBS showed a significant increase of BST-1, REX-1 and ABCG2 expression compared to 10% HS. In conclusion, HS promotes ASCs proliferation and viability but its ability to support the stemness property of ASCs was inferior to FBS.     

Identification of troponin I and actin, alpha cardiac muscle 1 as potential biomarkers for hearts of electrically stimulated chickens

Methods  

In this study, proteomics methods have been used to study the effects of different currents and voltages used to stun chickens. Protein profiles of chicken hearts were constructed to detect differences in protein expression and modification. The different voltages studied were 10 V, 40 V and 70 V, while the currents examined were 0.25 A, 0.5 A, and 0.75 A. The profiles obtained from these stunning conditions were compared to the non-stunned (0 A, 0 V) sample.     

Simultaneous determination of amlodipine and metoprolol in their combined dosage form using a synchronous fluorescence spectrofluorimetric method

Highly sensitive, rapid, accurate and precise synchronous fluorescence spectrofluorimetric method has been developed for simultaneous analysis of a mixture of amlodipine (AMD) and metoprolol (MET). The method relies on measuring the relative synchronous fluorescence intensity of both drugs at Δλ of 90 nm in acetate buffer solution at pH 5. The experimental parameters influencing the developed method were investigated and optimized. The method was linear over the ranges 0.2–2 μg/ml and 0.5–10 μg/ml for AMD and MET, respectively. The limits of detection were 50 ng/ml for AMD and 130 ng/ml for MET while the limits of quantitation were 150 ng/ml for AMD and 390 ng/ml for MET. The developed method was applied successfully for the determination of the two drugs in their co‐formulated tablet. The mean percent recoveries were found to be 100.51 and 99.57 for AMD and MET, respectively.     

MMP‐3 (–1171 5A/6A; Lys45Glu) variants affect serum levels of matrix metalloproteinase (MMP)‐3 and correlate with severity of COPD: A study of MMP‐3, MMP‐7 and MMP‐12 in a Tunisian population

Background

The present study aimed to examine the role of matrix metalloproteinase (MMP)‐3 [(–1171) 5A/6A; Lys45Glu (A/G)], MMP‐7 [(–181) A/G] and MMP‐12 [(–82) A/G; Asn357Ser (A/G)] variants in the development and severity of chronic obstructive pulmonary disease (COPD) in Tunisians.

Methods

Plethysmography was performed in all participants to measure forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC) and FEV1/FVC parameters. Genotyping of MMP‐3, MMP‐7 and MMP‐12 polymorphisms was carried out in 138 patients with COPD and 216 healthy controls using a polymerase chain reaction–restriction fragment length polymorphism. Serum levels of MMPs and cytokines (interleukin‐6, tumor necrosis factor‐α) were determined by an enzyme‐linked immunosorbent assay.

Results

No significant correlations were observed between genetic variations in MMP‐3, MMP‐7 and MMP‐12 and the risk of development of COPD. Additionally, no impact of MMP‐7 (–181) A/G and MMP‐12 [(–82) A/G; Asn357Ser (A/G)] polymorphisms was observed on the respective protein levels and clinical parameters of the disease. Interestingly, both MMP‐3 (–1171) 5A/6A and Lys45Glu (A/G) variants were associated with respiratory function, as well as with serum levels of MMP‐3 in COPD patients. A relationship was found between the (–1171) 6A and 45Glu (G) alleles of the MMP‐3 gene and enhanced airflow limitation among COPD patients. Additionally, carriers of the 6A6A and 45 GG genotypes present higher MMP‐3 levels than noncarriers.

Conclusions

MMP‐3 (–1171) 5A/6A and Lys45Glu (A/G) polymorphisms were associated with the decline of lung function among COPD patients. These results could be linked to the upregulation of MMP‐3 in serum from COPD patients carrying the (–1171) 6A and 45 G homozygous genotypes.     

Demyelination-Induced Inflammation Attracts Newly Born Neurons to the White Matter

There is compelling evidence that microglial activation negatively impacts neurogenesis. However, microglia have also been shown to promote recruitment of newly born neurons to injured areas of the gray matter. In the present study, we explored whether demyelination-triggered inflammation alters the process of neurogenesis in the white matter. A 2-μl solution of 0.04 % ethidium bromide was stereotaxically injected into the corpus callosum of adult male rats. Brain inflammation was dampened by daily injections of progesterone (5 mg/kg, s.c.) for 14 days. Control rats received oil (s.c.). Newly born neurons (DCX and Tbr2), microglia (Iba-1), astrocytes (vimentin or GFAP), oligodendrocyte progenitor cells (OPCs; NG2), and mature oligodendrocytes (CC-1) were monitored in the vicinity of demyelination site using immunofluorescent staining. Western blot was used to explore microglial polarization using M1 (iNOS) and M2 (arginase-1) markers. Focal demyelination elicited strong microglial and astroglial activation and reduced the number of OPCs at the site of demyelination. This inflammatory response was associated with enhanced number of newly born neurons in the white matter and the subventricular zone (SVZ). A proportion of newly born neurons within the white matter showed features of OPCs. Interestingly, blunting brain inflammation led to reduced neurogenesis around the demyelination area and in the SVZ. These data suggest that the white matter inflammation creates a conducive environment for the recruitment of newly born neurons. The fact that a sizable fraction of these newly born neurons adopt OPC features suggests that they could contribute to the remyelination process.     

Effects of aqueous cinnamon extract on chemically-induced carcinoma of hamster cheek pouch mucosa

This study aimed to investigate the effects of aqueous cinnamon extract (ACE) on 7, 12-Dimethylbenz[a]anthracene (DMBA)-induced oral carcinogenesis in hamster cheek pouch (HCP) mucosa. Sixty male Syrian hamsters were randomly divided into six equal groups. The hamsters of groups I, II and III received no treatment, DMBA and ACE respectively, for 16 weeks. Groups IV and V were handled as group II and concomitantly treated with ACE for the same period and additionally group V received ACE for other 16 weeks after the stoppage of DMBA application. Group VI hamsters were handled as group III and additionally received DMBA for other 16 weeks after the stoppage of ACE supplementation. Hamsters of each group were euthanized according to the experimental schedule. The buccal pouches were and prepared for H&E stain, PAS reagent, CD3 and PDGF immunohistochemical reactivity. All groups showed dysplastic changes with varying degrees except groups I and III. Deep invasive carcinomas were recorded in 90% of the samples of group II, 60% of group IV, 50% of group V and 40% of group VI. From the previous results, it can be concluded that ACE has the potentiality preventing oral cancer initiation better than inhibiting oral cancer progression.