Human-Derived Physiological Heat Shock Protein 27 Complex Protects Brain after Focal Cerebral Ischemia in Mice

Although challenging, neuroprotective therapies for ischemic stroke remain an interesting strategy for countering ischemic injury and suppressing brain tissue damage. Among potential neuroprotective molecules, heat shock protein 27 (HSP27) is a strong cell death suppressor. To assess the neuroprotective effects of HSP27 in a mouse model of transient middle cerebral artery occlusion, we purified a “physiological” HSP27 (hHSP27) from normal human lymphocytes. hHSP27 differed from recombinant HSP27 in that it formed dimeric, tetrameric, and multimeric complexes, was phosphorylated, and contained small amounts of αβ-crystallin and HSP20. Mice received intravenous injections of hHSP27 following focal cerebral ischemia. Infarct volume, neurological deficit scores, physiological parameters, and immunohistochemical analyses were evaluated 24 h after reperfusion. Intravenous injections of hHSP27 1 h after reperfusion significantly reduced infarct size and improved neurological deficits. Injected hHSP27 was localized in neurons on the ischemic side of the brain. hHSP27 suppressed neuronal cell death resulting from cytochrome c-mediated caspase activation, oxidative stress, and inflammatory responses. Recombinant HSP27 (rHSP27), which was artificially expressed and purified from Escherichia coli, and dephosphorylated hHSP27 did not have brain protective effects, suggesting that the phosphorylation of hHSP27 may be important for neuroprotection after ischemic insults. The present study suggests that hHSP27 with posttranslational modifications provided neuroprotection against ischemia/reperfusion injury and that the protection was mediated through the inhibition of apoptosis, oxidative stress, and inflammation. Intravenously injected human HSP27 should be explored for the treatment of acute ischemic strokes.     

Generation of Rejuvenated Antigen-Specific T Cells by Reprogramming to Pluripotency and Redifferentiation

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Highlights? Reprogramming of antigen-specific T cells to generate iPSCs (T-iPSCs) ? Redifferentiation of CD8⁺ T cells, with original antigen specificity, from T-iPSCs ? Newly differentiated T cells show high proliferation and elongated telomeres ? T cell antigen-specific cytotoxicity is maintained     

Oligodendrocyte Precursor Cells Support Blood-Brain Barrier Integrity via TGF-β Signaling

Trophic coupling between cerebral endothelium and their neighboring cells is required for the development and maintenance of blood-brain barrier (BBB) function. Here we report that oligodendrocyte precursor cells (OPCs) secrete soluble factor TGF-β1 to support BBB integrity. Firstly, we prepared conditioned media from OPC cultures and added them to cerebral endothelial cultures. Our pharmacological experiments showed that OPC-conditioned media increased expressions of tight-junction proteins and decreased in vitro BBB permeability by activating TGB-β-receptor-MEK/ERK signaling pathway. Secondly, our immuno-electron microscopic observation revealed that in neonatal mouse brains, OPCs attach to cerebral endothelial cells via basal lamina. And finally, we developed a novel transgenic mouse line that TGF-β1 is knocked down specifically in OPCs. Neonates of these OPC-specific TGF-β1 deficient mice (OPC-specific TGF-β1 partial KO mice: Pdgfra^Cre/Tgfb1^flox/wt mice or OPC-specific TGF-β1 total KO mice: Pdgfra^Cre/Tgfb1^flox/flox mice) exhibited cerebral hemorrhage and loss of BBB function. Taken together, our current study demonstrates that OPCs increase BBB tightness by upregulating tight junction proteins via TGF-β signaling. Although astrocytes and pericytes are well-known regulators of BBB maturation and maintenance, these findings indicate that OPCs also play a pivotal role in promoting BBB integrity.     

Limitations on tree seedling establishment across ecotones between abandoned fields and adjacent broad-leaved forests in eastern Japan

Field abandonment has increased over several decades under a long-term recession in Japanese agriculture. To support effective farmland management or reforestation, the patterns of tree recruitment in abandoned fields, which influence management costs, need to be clarified. We investigated tree seedling establishment and microsite variables along forest–field transects in 11 abandoned fields adjacent to secondary broad-leaved forests in the eastern Kanto region of Japan. Generalized linear mixed model analysis indicated that tree seedling establishment in abandoned fields was not correlated with the period of abandonment. Instead, it was positively correlated with vegetation openness. The dominance of a woody vine (Pueraria lobata), an annual liana (Humulus japonicus), and a dwarf bamboo (Pleioblastus chino) mainly explained low vegetation openness within the fields. In addition, the establishment of evergreen tree seedlings and tree seedlings dispersed by hoarding was negatively correlated with the distance from the forest edge. In the forest interior, taller understory vegetation at the edge correlated with lower seedling establishment. These results suggest that seedling establishment across forest–field ecotones is restricted by the dominance of competitive natives within abandoned fields and distance-dependent limitations in both abandoned fields and adjacent forest.     

Allelopathic interference of sweet potato with cogongrass and relevant species

Cogongrass (Imperata cylindria) is an invasive weed and harmful to ecological systems and agricultural production in many countries. It was found that plant extracts and root exudates of sweet potato (Ipomoea batatas) exhibit allelopathic potential and inhibit the growth of cogongrass to a greater extent than either barnyardgrass (Echinochloa crus-galli), Indian goose-grass (Eleushine indica), or lettuce (Lactuca sativa) in bioassays. Greenhouse trials indicated that sweet potato soil reduced the emergence of the noxious weed by 50 %, yet exhibited either weaker inhibition or the promotion of barnyardgrass, Bidens (Bidens pilosa), and Leucaena (Leucaena leucocephala), while the desired growth of upland rice (Oryza sativa) was not affected. In cogongrass fields, the incorporation of 1–2 tons aboveground parts and cultivation of sweet potato provided 80–85 % weed control. On the other hand, the reduction of congograss in fields may be offset by the alternate invasion of B. pilosa which multiplied its biomass by 2–6 times with sweet potato amended soils. The findings of this study indicate that sweet potato is an effective crop in the biologic management of the invasive cogongrass in agricultural fields, thus the interactive mechanism between sweet potato and the invasive weed demands further investigation. Ecologically, this study highlights the specificity of allelopathic interactions between cogongrass and sweet potato that is helpful to minimize the disturbance from infestation of this invasive weed against native species and crops.     

Novel, objective, multivariate biomarkers composed of plasma amino acid profiles for the diagnosis and assessment of inflammatory bowel disease

Background

Inflammatory bowel disease (IBD) is a chronic intestinal disorder that is associated with a limited number of clinical biomarkers. In order to facilitate the diagnosis of IBD and assess its disease activity, we investigated the potential of novel multivariate indexes using statistical modeling of plasma amino acid concentrations (aminogram).

Methodology and Principal Findings

We measured fasting plasma aminograms in 387 IBD patients (Crohn''s disease (CD), n = 165; ulcerative colitis (UC), n = 222) and 210 healthy controls. Based on Fisher linear classifiers, multivariate indexes were developed from the aminogram in discovery samples (CD, n = 102; UC, n = 102; age and sex-matched healthy controls, n = 102) and internally validated. The indexes were used to discriminate between CD or UC patients and healthy controls, as well as between patients with active disease and those in remission. We assessed index performances using the area under the curve of the receiver operating characteristic (ROC AUC). We observed significant alterations to the plasma aminogram, including histidine and tryptophan. The multivariate indexes established from plasma aminograms were able to distinguish CD or UC patients from healthy controls with ROC AUCs of 0.940 (95% confidence interval (CI): 0.898–0.983) and 0.894 (95%CI: 0.853–0.935), respectively in validation samples (CD, n = 63; UC, n = 120; healthy controls, n = 108). In addition, other indexes appeared to be a measure of disease activity. These indexes distinguished active CD or UC patients from each remission patients with ROC AUCs of 0.894 (95%CI: 0.853–0.935) and 0.849 (95%CI: 0.770–0.928), and correlated with clinical disease activity indexes for CD (r_s = 0.592, 95%CI: 0.385–0.742, p<0.001) or UC (r_s = 0.598, 95%CI: 0.452–0.713, p<0.001), respectively.

Conclusions and Significance

In this study, we demonstrated that established multivariate indexes composed of plasma amino acid profiles can serve as novel, non-invasive, objective biomarkers for the diagnosis and monitoring of IBD, providing us with new insights into the pathophysiology of the disease.     

Dynamic Changes in the Spatiotemporal Localization of Rab21 in Live RAW264 Cells during Macropinocytosis

Rab21, a member of the Rab GTPase family, is known to be involved in membrane trafficking, but its implication in macropinocytosis is unclear. We analyzed the spatiotemporal localization of Rab21 in M-CSF-stimulated RAW264 macrophages by the live-cell imaging of fluorescent protein-fused Rab21. It was demonstrated that wild-type Rab21 was transiently associated with macropinosomes. Rab21 was recruited to the macropinosomes after a decrease in PI(4,5)P₂ and PI(3,4,5)P₃ levels. Although Rab21 was largely colocalized with Rab5, the recruitment of Rab21 to the macropinosomes lagged a minute behind that of Rab5, and preceded that of Rab7. Then, Rab21 was dissociated from the macropinosomes prior to the accumulation of Lamp1, a late endosomal/lysosomal marker. Our analysis of Rab21 mutants revealed that the GTP-bound mutant, Rab21-Q78L, was recruited to the macropinosomes, similarly to wild-type Rab21. However, the GDP-bound mutant, Rab21-T33N, did not localize on the formed macropinosomes, suggesting that the binding of GTP to Rab21 is required for the proper recruitment of Rab21 onto the macropinosomes. However, neither mutation of Rab21 significantly affected the rate of macropinosome formation. These data indicate that Rab21 is a transient component of early and intermediate stages of macropinocytosis, and probably functions in macropinosome maturation before fusing with lysosomal compartments.     

State and management of wetlands in Bangladesh

Wetlands are a vital link between land and water in Bangladesh. A majority of the people of Bangladesh are critically dependent on wetlands. In this paper, the values of wetlands, causes and effects of wetlands degradation, as well as the present wetlands management approach, are analyzed and recommendations for wetlands management are suggested based on participatory rural appraisal (PRA), field visit, personal experience, and existing literature and information. Wetlands play a crucial role in maintaining the ecological balance of ecosystems, but wetlands habitat of Bangladesh is under constant threat due to increase of population, intensive agriculture, overfishing, siltation, pollution, ill-planned infrastructures, lack of institutional coordination, lack of awareness, etc. As a result biodiversity is reducing, many species of flora and fauna are threatened, wetlands-based ecosystem is degenerating, and the living conditions of local people are deteriorating as livelihoods, socioeconomic institutions, and cultural values are affected. Wetlands management is not addressed separately in water management activities of Bangladesh. In order to balance human needs and wetlands conservation, a mainly community-based wetlands management approach has been taken in Bangladesh, but this is not enough to prevent the degradation of wetlands. Therefore, Bangladesh now needs a comprehensive strategy combining political, economic, social, and technological approaches to stop further degradation of wetlands. Therefore, wetlands management should be incorporated into a system of integrated land and water use and, indeed, into the socioeconomic system of the country. Policies, strategies, and management plans for sustainable use and conservation of wetlands of Bangladesh must be based on solid knowledge and understanding of their ecological and socioeconomic functions and processes.