Given recent advances in automated oligosaccharide synthesis, analytical techniques that can be coupled to a synthetic framework are needed to not just identify but also purify to homogeneity ...protected carbohydrate compounds at levels of ≥99.5% purity. Herein, an alternate-pump recycling high-performance liquid chromatography (R-HPLC) method has been developed to allow purification of protected carbohydrates at levels of ≥99.5% purity.
CDK5 plays an indispensable role in the central nervous system, and its deregulation is involved in neurodegeneration. We report the crystal structure of a complex between CDK5 and p25, a fragment of ...the p35 activator. Despite its partial structural similarity with the cyclins, p25 displays an unprecedented mechanism for the regulation of a cyclin-dependent kinase. p25 tethers the unphosphorylated T loop of CDK5 in the active conformation. Residue Ser159, equivalent to Thr160 on CDK2, contributes to the specificity of the CDK5-p35 interaction. Its substitution with threonine prevents p35 binding, while the presence of alanine affects neither binding nor kinase activity. Finally, we provide evidence that the CDK5-p25 complex employs a distinct mechanism from the phospho-CDK2-cyclin A complex to establish substrate specificity.
Morphological and ecophysiological traits showed by male and female
Populus tomentosa
Carr. trees were studied under various degrees of water and alkaline stresses. The results showed that different ...adaptations to drought and alkaline stresses were adopted by each gender; males possessed a much higher tolerance to both stresses compared to females. In contrast to females, the males exhibited a lower inhibition in total biomass, total leaf area, net photosynthetic rate, stomatal conductance, leaf carbon and nitrogen concentrations as well as water- and nitrogen-use efficiency in response to drought and alkaline stresses. Nevertheless, compared to the males, the females showed a higher plasticity in root biomass/shoot biomass ratio, fine root/coarse root ratio, and intrinsic water-use efficiency, indicating that the males and females differed in some of trade-offs between growth and stress defence to maximize water and nitrogen gains under both stress conditions.
We developed a path imbalance measuring system using a reference interferometer with alterable optical path difference (OPD), aiming to eliminate the uncertainties due to synthetic wavelength ...measurement and remove the requirement of a known and stable reference OPD in frequency scanning interferometry. The path imbalance can be solved by using the phase ratios between the two interferometers produced before and after altering the OPD in the reference interferometer. The results have shown that the measurement uncertainty and the path imbalance are linearly related and a combined relative uncertainty of 4.9 × 10−6 (1σ) in path imbalance measurement over a range from 0.5 m to 50 m is achieved. Besides, we analyzed the contributions to the uncertainty that limit the performance of the system, and we discussed how to obtain a better measurement uncertainty.
Peng L, Ren LB, Dong G, Wang CL, Xu P, Ye L, Zhou XD. Wnt5a promotes differentiation of human dental papilla cells. International Endodontic Journal, 43, 404–412, 2010.
Aim To investigate the role ...of Wnt5a in the process of differentiation of human dental papilla cells (HDPCs).
Methodology Recombinant adenovirus encoding full‐length Wnt5a cDNA was constructed to investigate the biological role of Wnt5a on the differentiation of HDPCs. The effect of Wnt5a on HDPCs differentiation was determined by ALP activity assay, ALP staining and mineral induction assay. Mineralization‐related gene expressions were assessed by RT‐PCR.
Results Immunostaining revealed Wnt5a expression in the odontoblast layer and dental papilla tissue. Over‐expression of Wnt5a by transfecting HDPCs with an Wnt5a‐carrying construct increased ALPase activity and the formation of mineralized nodules of HDPCs. RT‐PCR analysis showed that the expressions of mineralization‐related genes, such as bone sialoprotein, collagen type I, osteonectin, osteopontin (OCN), dentine matrix protein‐1 were up‐regulated by Wnt5a.
Conclusions Wnt5a promoted differentiation of HDPCs.
Rocio virus (ROCV) is a highly neuropathogenic mosquito-transmitted flavivirus responsible for an unprecedented outbreak of human encephalitis during 1975-1976 in Sao Paulo State, Brazil. Previous ...studies have shown an increased number of inflammatory macrophages in the central nervous system (CNS) of ROCV-infected mice, implying a role for macrophages in the pathogenesis of ROCV. Here, we show that ROCV infection results in increased expression of CCL2 in the blood and in infiltration of macrophages into the brain. Moreover, we show, using CCR2 knockout mice, that CCR2 expression is essential for macrophage infiltration in the brain during ROCV infection and that the lack of CCR2 results in increased disease severity and mortality. Thus, our findings show the protective role of CCR2-mediated infiltration of macrophages in the brain during ROCV infection.
To ascertain the ability of adrenomedullin (ADM) and proadrenomedullin (proADM) to predict mortality in sepsis patients.
A systematic literature search was made of the PubMed, EMBASE, Cochrane and ...China National Knowledge Infrastructure (CNKI) databases before May 2017, supplemented by manual searches of references. A meta-analysis of high-quality clinical studies was subsequently performed to assess the association between ADM/proADM and mortality risk among patients with sepsis.
Thirteen studies involving 2556 patients were included in the study.
Two reviewers independently identified articles, extracted data, assessed quality and cross-checked the results. The predictive values of ADM and proADM referred to mortality were assessed by relative risk (RR). The overall diagnostic accuracy of ADM and proADM in application to sepsis was pooled according to a bivariate model. Publication bias was assessed using Deek's funnel plot asymmetry test.
Elevated ADM or proADM levels were associated with increased mortality (pooled RR=3.31; 95%CI 2.31-4.75). Subgroup analyses indicated the pooled RRs were 3.12 (95%CI 1.75-5.56) and 3.43 (95%CI 2.21-5.31) for ADM and proADM, respectively. The pooled sensitivity and specificity were 0.72 (95%CI 0.64-0.78) and 0.77 (95%CI 0.69-0.83), respectively. The overall area under the summary receiver operating characteristic (SROC) curve was 0.80 (95%CI 0.77-0.84). Publication bias was not statistically significant.
Both ADM and proADM might serve as useful markers for predicting the prognosis of sepsis.
Proteins that communicate signals from the cytoskeleton to the nucleus are prime targets for effectors of metastasis as they often transduce signals regulating adhesion, motility, and invasiveness. ...LIM domain proteins shuttle between the cytoplasm and the nucleus, and bind to partners in both compartments, often coupling changes in gene expression to extracellular cues. In this work, we characterize LIMD2, a mechanistically undefined LIM-only protein originally found to be overexpressed in metastatic lesions but absent in the matched primary tumor. LIMD2 levels in fresh and archival tumors positively correlate with cell motility, metastatic potential, and grade, including bladder, melanoma, breast, and thyroid tumors. LIMD2 directly contributes to these cellular phenotypes as shown by overexpression, knockdown, and reconstitution experiments in cell culture models. The solution structure of LIMD2 that was determined using nuclear magnetic resonance revealed a classic LIM-domain structure that was highly related to LIM1 of PINCH1, a core component of the integrin-linked kinase-parvin-pinch complex. Structural and biochemical analyses revealed that LIMD2 bound directly to the kinase domain of integrin-linked kinase (ILK) near the active site and strongly activated ILK kinase activity. Cells that were null for ILK failed to respond to the induction of invasion by LIMD2. This strongly suggests that LIMD2 potentiates its biologic effects through direct interactions with ILK, a signal transduction pathway firmly linked to cell motility and invasion. In summary, LIMD2 is a new component of the signal transduction cascade that links integrin-mediated signaling to cell motility/metastatic behavior and may be a promising target for controlling tumor spread.
To investigate whether brassinosteroids (BR) affects cytokinin (CK)-induced anthocyanin biosynthesis, seedlings of the
Arabidopsis dwarf4
(
dwf4
) mutants including
partially suppressing coi1
(
psc1
...) and
dwf4-102
, which are defective in the BR biosynthesis, and the
brassinosteroid-insensitive 1–4
(
bri1-4
) mutant defective in BR signalling were used for the analysis of CK-induced anthocyanin accumulation and the expression of anthocyanin biosynthetic genes and WD-repeat/Myb/bHLH transcription factors. The results show that the CK-induced anthocyanin accumulation was remarkably reduced in
dwf4
and
bri1-4
mutants, but distinctly increased in the wild type (WT) treated with BR. Moreover, the CK-induced expressions of the late anthocyanin biosynthetic genes including
dihydroflavonol reductase
,
leucoanthocyanidin dioxygenase
, and
UDP-glucose: flavonoid-3-O-glucosyl transferase
were significantly reduced in
bri1-4
and
dwf4-102
mutants compared to WT. In addition, the expressions of transcription factors
production of anthocyanin pigment 1
(
PAP1
),
glabra 3
(
GL3
), and
enhancer of glabra 3
(
EGL3
) were induced by CK in WT but not in the
bri1-4
and
dwf4-102
mutants. These results indicate that BR enhanced the CK-induced anthocyanin biosynthesis by up-regulating the late anthocyanin biosynthetic genes and this regulation might be mediated by the transcription factors
PAP1
,
GL3
, and
EGL3
.
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