Improved stability of methanolic Wright's stain solution with dual additives

Degradation of methanolic Wright's stain solutions was greatly diminished with the addition of diethylamine hydrochloride and dimethylamine hydrochloride as costabilizers. Precipitation problems were eliminated by the dual additives. The stabilized stain solutions demonstrated good staining performance on blood smears. Methods for predicting the shelf life using calculated analytical parameters are described. Using these methods, the shelf life of a control stain solution was predicted to be 0.7 years; predicted shelf life was more than tripled with the addition of diethylamine hydrochloride and was increased approximately 27 times with the addition of both diethylamine hydrochloride and dimethylamine hydrochloride.     

Metabolic engineering of Clostridium tyrobutyricum for enhanced butyric acid production with high butyrate/acetate ratio

Applied Microbiology and Biotechnology - Butyric acid fermentation by Clostridium couples with the synthesis of acetic acid. But the presence of acetic acid reduces butyric acid yield and increases...     

Different Glycosylation in Acetylcholinesterases from Mammalian Brain and Erythrocytes

Acetylcholinesterases (EC 3.1.1.7, AChE) have varying amounts of carbohydrates attached to the core protein. Sequence analysis of the known primary structures gives evidence for several asparagine-linked carbohydrates. From the differences in molecular mass determined on sodium dodecyl sulfate-polyacrylamide gel before and after deglycosylation with N-glycosidase F (EC 3.2.2.18), it is seen that dimeric AChE from red cell membranes is more heavily glycosylated than the tetrameric brain enzyme. Furthermore, dimeric and tetrameric forms of bovine AChE are more heavily glycosylated than the corresponding human enzymes. Monoclonal antibodies 2E6, 1H11, and 2G8 raised against detergent-soluble AChE from electric organs of Torpedo nacline timilei as well as Elec-39 raised against AChE from Electrophorus electricus cross-reacted with AChE from bovine and human brain but not with AChE from erythrocytes. Treatment of the enzyme with N-glycosidase F abolished binding of monoclonal antibodies, suggesting that the epitope, or part of it, consists of N-linked carbohydrates. Analysis of N-acetylglucosamine sugars revealed the presence of N-acetylglucosamine in all forms of cholinesterases investigated, giving evidence for N-linked glycosylation. On the other hand, N-acetylgalactosamine was not found in AChE from human and bovine brain or in butyrylcholinesterase (EC 3.1.1.8) from human serum, indicating that these forms of cholinesterase did not contain O-linked carbohydrates. Despite the notion that within one species, the different forms of AChE arise from one gene by different splicing, our present results show that dimeric erythrocyte and tetrameric brain AChE must undergo different postsynthetic modifications leading to differences in their glycosylation patterns.     

Effects of warming and nutrient fluctuation on invader Chromolaena odorata and natives in artificial communities

Plant Ecology - There is increasing evidence that climate change and nutrient fluctuations can affect the invasion of alien plants. However, most studies have been performed in pairwise experiments...     

Seasonal dynamics of soil fauna in the subalpine and alpine forests of west Sichuan at different altitudes

The climate (especially temperature) often plays an important role in the structure, function as well as composition of soil organisms in different latitudes and altitudes. As one of the essential components of soil ecosystem, soil faunal community not only lays their roles as soil engineer in material cycling and energy flow, but also acts as the sensitive bio-indicator to environmental change. However, little information has been available on the responses of soil faunal community to the changed environment at different altitudes and seasons. In order to understand the seasonal dynamics of soil faunal diversity under different forests with varying altitudes, three fir (Abies faxoniana) forests were selected covering a 600 m vertical transition zone. The primary fir forest at 3600 m (A1) of altitude, mixed fir and birch forest at 3300 m (A2) of altitude, and secondary fir forest at 3000 m (A3) of altitude are representative forests in the subalpine and alpine region of west Sichuan. A 2 years study was conducted in the three subalpine and alpine forests from May in 2009 until October in 2010. Soil samples were collected in both the soil organic layer and mineral soil layer. Soil macro-fauna were picked up by hand in the fields. Meso/micro-fauna and damp living fauna were separated and collected from the soil samples by Baermann and Tullgren methods in laboratory, respectively. A total of 74,827 individuals were collected in the 2 years, belonging to seven phyla, 16 classes, 31 orders and 125 families by preliminary identification. Similar dominant groups were detected in different forests at different altitudes, consisting of Spirostreptida, Formicidae, Staphylinidae, Hesperinidae, Onychiuridae, Isotomidae, Oribatuloidae, Alicoragiidae, Secernentea, and Adenophorea. In contrast, the ordinary species of macro-fauna and the ratios of Acarina to Collembolan were obviously different. For instance, the ordinary species were dominated by Cydmaenidae and Mycetophilidae at the A1, Scaphidiidae and Helicinidae at the A2, and Lumbricida and Agelenidae at the A3, respectively. Both the individual density and the number of soil faunal groups were significantly higher in soil organic layer than those in mineral soil layer. The density and group of macro-, meso- and micro-fauna in different forests showed the order as A2 > A1 > A3, but the density of damp living fauna showed the order as A1 > A2 > A3. The functional groups of macro-fauna were mainly dominated by saprozoic. The highest density and group of macro-fauna was observed in August, while the highest value of meso/micro-fauna was detected in October. In addition, the Jacard similarity indices showed that the composition and structure of soil fauna were similar in the different forests varied with altitudes, but the Shannon–Wiener indices were significantly different. The highest values of Shannon–Wiener indices were observed in October at both the A1 and A3, and in August at the A2. The results suggested that soil faunal community kept a high diversity in the subalpine and alpine forests of west Sichuan, and their structures were significantly affected by the variation of altitudes, which provided important scientific evidences for understanding the ecological processes in the subalpine and alpine coniferous forests.     

Hyperosmolarity enhanced susceptibility to renal tubular fibrosis by modulating catabolism of type I transforming growth factor‐β receptors

Hyperosmolarity plays an essential role in the pathogenesis of diabetic tubular fibrosis. However, the mechanism of the involvement of hyperosmolarity remains unclear. In this study, mannitol was used to evaluate the effects of hyperosmolarity on a renal distal tubule cell line (MDCK). We investigated transforming growth factor‐β receptors and their downstream fibrogenic signal proteins. We show that hyperosmolarity significantly enhances the susceptibility to exogenous transforming growth factor (TGF)‐β1, as mannitol (27.5 mM) significantly enhanced the TGF‐β1‐induced increase in fibronectin levels compared with control experiments (5.5 mM). Specifically, hyperosmolarity induced tyrosine phosphorylation on TGF‐β RII at 336 residues in a time (0–24 h) and dose (5.5–38.5 mM) dependent manner. In addition, hyperosmolarity increased the level of TGF‐β RI in a dose‐ and time‐course dependent manner. These observations may be closely related to decreased catabolism of TGF‐β RI. Hyperosmolarity significantly downregulated the expression of an inhibitory Smad (Smad7), decreased the level of Smurf 1, and reduced ubiquitination of TGF‐β RI. In addition, through the use of cycloheximide and the proteasome inhibitor MG132, we showed that hyperosmolarity significantly increased the half‐life and inhibited the protein level of TGF‐β RI by polyubiquitination and proteasomal degradation. Taken together, our data suggest that hyperosmolarity enhances cellular susceptibility to renal tubular fibrosis by activating the Smad7 pathway and increasing the stability of type I TGF‐β receptors by retarding proteasomal degradation of TGF‐β RI. This study clarifies the mechanism underlying hyperosmotic‐induced renal fibrosis in renal distal tubule cells. J. Cell. Biochem. 109: 663–671, 2010. © 2010 Wiley‐Liss, Inc.     

Cardiac telocytes exist in the adult Xenopus tropicalis heart

Recent research has revealed that cardiac telocytes (CTs) play an important role in cardiac physiopathology and the regeneration of injured myocardium. Recently, we reported that the adult Xenopus tropicalis heart can regenerate perfectly in a nearly scar‐free manner after injury via apical resection. However, whether telocytes exist in the X tropicalis heart and are affected in the regeneration of injured X tropicalis myocardium is still unknown. The present ultrastructural and immunofluorescent double staining results clearly showed that CTs exist in the X tropicalis myocardium. CTs in the X tropicalis myocardium were mainly twined around the surface of cardiomyocyte trabeculae and linked via nanocontacts between the ends of the telopodes, forming a three‐dimensional network. CTs might play a role in the regeneration of injured myocardium.     

Gastrodin relieves inflammation injury induced by lipopolysaccharides in MRC‐5 cells by up‐regulation of miR‐103

The beneficial function of gastrodin towards many inflammatory diseases has been identified. This study designed to see the influence of gastrodin in a cell model of chronic obstructive pulmonary disease (COPD). MRC‐5 cells were treated by LPS, before which gastrodin was administrated. The effects of gastrodin were evaluated by conducting CCK‐8, FITC‐PI double staining, Western blot, qRT‐PCR and ELISA. Besides this, the downstream effector and signalling were studied to decode how gastrodin exerted its function. And dual‐luciferase assay was used to detect the targeting link between miR‐103 and lipoprotein receptor‐related protein 1 (LRP1). LPS induced apoptosis and the release of MCP‐1, IL‐6 and TNF‐α in MRC‐5 cells. Pre‐treating MRC‐5 cells with gastrodin attenuated LPS‐induced cell damage. Meanwhile, p38/JNK and NF‐κB pathways induced by LPS were repressed by gastrodin. miR‐103 expression was elevated by gastrodin. Further, the protective functions of gastrodin were attenuated by miR‐103 silencing. And LRP1 was a target of miR‐103 and negatively regulated by miR‐103. The in vitro data illustrated the protective function of gastrodin in LPS‐injured MRC‐5 cells. Gastrodin exerted its function possibly by up‐regulating miR‐103 and modulating p38/JNK and NF‐κB pathways.     

A stroma-related lncRNA panel for predicting recurrence and adjuvant chemotherapy benefit in patients with early-stage colon cancer

The heterogeneity in prognoses and chemotherapeutic responses of colon cancer patients with similar clinical features emphasized the necessity for new biomarkers that help to improve the survival prediction and tailor therapies more rationally and precisely. In the present study, we established a s troma-related l ncRNA s ignature (SLS) based on 52 lncRNAs to comprehensively predict clinical outcome. The SLS model could not only distinguish patients with different recurrence and mortality risks through univariate analysis, but also served as an independent factor for relapse-free and overall survival. Compared with the conventionally used TNM stage system, the SLS model clearly possessed higher predictive accuracy. Moreover, the SLS model also effectively screened chemotherapy-responsive patients, as only patients in the low-SLS group could benefit from adjuvant chemotherapy. The following cell infiltration and competing endogenous RNA (ceRNA) network functional analyses further confirmed the association between the SLS model and stromal activation-related biological processes. Additionally, this study also identified three phenotypically distinct colon cancer subtypes that varied in clinical outcome and chemotherapy benefits. In conclusion, our SLS model may be a significant determinant of survival and chemotherapeutic decision-making in colon cancer and may have a strong clinical transformation value.