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Re-excavation of the Peking man site at Zhoukoudian was initiated in 2009. A taphonomic analysis of the skeletal remains of Cervus ( Sika ) grayi , a dominant species from Layer 3 of the extant West Section of the site strongly negates a role played by hominins in the accumulation and modification of the bones. Instead, it is argued here that some individuals of this medium-sized deer species were probably trapped in the cave by accident and eventually preserved in situ in the Quaternary sediments. The virtual absence of stone artifacts and human skeletal remains from the 2009–2010 field seasons has further strengthened this conclusion. Bearing in mind evidence of a series of cave collapse episodes, we tentatively propose that the cave morphology during the formation of Layer 3 at the West Section was probably inhospitable to Homo erectus pekinensis . [ABSTRACT FROM AUTHOR]

Lead (Pb) is a toxic element which is released as a result of anthropogenic activities, and Pb stable isotope ratios provide a means to distinguish sources and transport pathways in receiving environments. In this study, isotopes of bioaccumulated Pb (204Pb, 206Pb, 207Pb, 208Pb) were examined for diverse terrestrial and aquatic biota from three areas in western Canada: (a) otter, marten, gulls, terns, and wood frogs in the Alberta Oil Sands Region (AOSR), (b) fish, plankton, and gulls of Great Slave Lake (Yellowknife, Northwest Territories), and (c) wolverine from the Yukon. Aquatic and terrestrial biota from different habitats and a broad geographic area showed a remarkable similarity in their Pb isotope composition (grand mean ± 1 standard deviation: 206Pb/207Pb = 1.189 ± 0.007, 208Pb/207Pb = 2.435 ± 0.009, n = 116). Comparisons with Pb isotope ratios of local sources and environmental receptors showed that values in biota were most similar to those of atmospheric Pb, either measured in local aerosols influenced by industrial activities in the AOSR or in lichens (an aerosol proxy) near Yellowknife and in the Yukon. Biotic Pb isotope ratios were different from those of local geogenic Pb. Although the Pb isotope measurements could not unambiguously identify the specific anthropogenic sources of atmospheric Pb in biota, initial evidence points to the importance of fossil fuels currently used in transportation and power generation. Further research should characterize bioavailable chemical species of Pb in aerosols and important emission sources in western Canada. [Display omitted] • Pb isotope ratios were strikingly similar in diverse animals from western Canada. • The bioaccumulated Pb originated primarily from atmospheric aerosols. • Natural geogenic sources contributed little to bioaccumulated Pb. • Pb isotopes suggest aerosol Pb is highly bioavailable. • Fossil fuels may be important sources of bioavailable Pb to fish and wildlife. [ABSTRACT FROM AUTHOR]

This paper proposes a new gravitational search algorithm to solve the unit commitment (UC) problem, which is integrated binary gravitational search algorithm (BGSA) with the Lambda-iteration method. The proposed method is enhanced by priority list based on the unit characteristics and heuristic search strategies to repair the spinning reserve and minimum up/down time constraints. Furthermore, local mutation strategies are applied to improve the performance of BGSA. The implementation of the proposed method for UC problem consists of three stages. Firstly, the BGSA based on priority list is applied for solution unit scheduling when neglecting minimum up/down time constraints. Secondly, heuristic search strategies are used to handle minimum up/down time constraints and decommit excess spinning reserve units. Thirdly, local mutation strategies are raised to avoid premature convergence of the algorithm and prevent it from trapping into local optima. Finally, Lambda-iteration method is adopted to solve economic load dispatch based on the obtained unit schedule. The feasibility and effectiveness of the proposed method is verified by the systems with the number of units in the range of 10–100 and the results are compared with those of other methods reported in literatures. The results clearly show that the proposed method gives better quality solutions than other methods. [ABSTRACT FROM AUTHOR]

Abstract: The Tianyuan cave near Zhoukoudian is among the most special localities in North China for its many porcupine gnawing marks on the bones, as is often the case acknowledged only in South China. But porcupines did not gnaw all the bones, that is gnawing is selective. Skulls, mandibles, tooth roots and long bones are frequently gnawed. The short bones are almost free from gnawing. The selection indicates that, for porcupines, bone gnawing is not only a necessity for attrition of incisors, but also a process of extracting nutrients from the marrow. Additionally, the porcupine gnawing marks can also be employed to determine the provenance of the fossils that were discovered by the local people before the systematic excavation. Those bearing porcupine tooth marks only appear in the upper assemblage. The other significance of the gnawing marks is to infer the habitant of the cave. The bones from the human fossil bearing layer have few tooth marks, which may represent a period of human occupation of the cave. In the upper assemblage, the gnawing marks appear frequently, it may indicate that porcupine and other rodents lived in the cave during this time span. [Copyright &y& Elsevier]

Abstract: A fusion between gene encoding fluoresce-enhanced green fluorescent protein variant (EGFP) and soluble domain of human B-cell-activating factor of the TNF family (sBAFF) was constructed and expressed in Escherichia coli. The EGFP/sBAFF had an apparent molecular weight of 45 kDa and was detected with anti-hsBAFF and anti-His6 monoclonal antibodies. After being purified by immobilized metal affinity chromatography (IMAC), the fusion protein retained similar fluorescence spectra to those of EGFP. Biological activity assays showed the EGFP/sBAFF as well as sBAFF could co-stimulated human B lymphocyte proliferation in vitro. In addition, EGFP/sBAFF has shown specific binding to BAFF receptors positive-cells and the stained cells could be analyzed with flow cytometry. Thus, the fusion protein represents a readily obtainable source of biologically active sBAFF that may prove useful in further studies on BAFF and its receptors. [Copyright &y& Elsevier]

• Rapamycin attenuation of BAFF-elevated [Ca2+] i links to its inhibition of B-cell proliferation/survival. • Rapamyicn blunts mTORC1/2-mediated [Ca2+] i elevations induced by BAFF. • Rapamyicn inhibits BAFF-stimulated B-cell proliferation/survival by blocking Ca2+-CaMKII-dependent PTEN/Akt-Erk1/2 signaling. • Rapamycin may be an effective agent for prevention of BAFF-induced aggressive or neoplastic B-cell disorders. B-cell activating factor (BAFF) is a crucial survival factor for B cells, and excess BAFF contributes to development of autoimmune diseases. Recent studies have shown that rapamycin can prevent BAFF-induced B-cell proliferation and survival, but the underlying mechanism remains to be elucidated. Here we found that rapamycin inhibited human soluble BAFF (hsBAFF)-stimulated cell proliferation by inducing G 1 -cell cycle arrest, which was through downregulating the protein levels of CDK2, CDK4, CDK6, cyclin A, cyclin D1, and cyclin E. Rapamycin reduced hsBAFF-stimulated cell survival by downregulating the levels of anti-apoptotic proteins (Mcl-1, Bcl-2, Bcl-xL and survivin) and meanwhile upregulating the levels of pro-apoptotic proteins (BAK and BAX). The cytostatic and cytotoxic effects of rapamycin linked to its attenuation of hsBAFF-elevated intracellular free Ca2+ ([Ca2+] i). In addition, rapamycin blocked hsBAFF-stimulated B-cell proliferation and survival by preventing hsBAFF from inactivating PTEN and activating the Akt-Erk1/2 pathway. Overexpression of wild type PTEN or ectopic expression of dominant negative Akt potentiated rapamycin's suppression of hsBAFF-induced Erk1/2 activation and proliferation/viability in Raji cells. Interestingly, PP242 (mTORC1/2 inhibitor) or Akt inhibitor X, like rapamycin (mTORC1 inhibitor), reduced the basal or hsBAFF-induced [Ca2+] i elevations. Chelating [Ca2+] i with BAPTA/AM, preventing [Ca2+] i elevation using EGTA, 2-APB or verapamil, inhibiting CaMKII with KN93, or silencing CaMKII strengthened rapamycin's inhibitory effects. The results indicate that rapamycin inhibits BAFF-stimulated B-cell proliferation and survival by blunting mTORC1/2-mediated [Ca2+] i elevations and suppressing Ca2+-CaMKII-dependent PTEN/Akt-Erk1/2 signaling pathway. Our finding underscores that rapamycin may be exploited for prevention of excessive BAFF-induced aggressive B-cell malignancies and autoimmune diseases. [ABSTRACT FROM AUTHOR]

The process of the Middle to Upper Palaeolithic transition has been hotly debated for many years, but East Asia has not been a significant part of that debate until recently. The Shuidonggou (SDG) site complex, yielding Initial Upper Paleolithic (IUP) blade-rich assemblages, has been given more and more importance since its discovery and first excavation in the 1920s. This paper presents a specific study of the archaeological remains yielded by three years of systematic, modern excavations at Shuidonggou locality 7 (SDG7), an important locality within the SDG site cluster. Although the archaeological deposits of SDG7 had undergone some hydraulic disturbances, detailed lithic analyses identified two distinct technological systems. One is termed autochthonous and is a predominantly flake-tool technology, and the other is an allochthonous Levallois-like technology which coexisted within the same Lower Concentrated Layer for some time before disappearing, implying the potential existence of technological diffusion and interaction between the two different technologies. The technological variability and change suggested by the evidence from SDG7 reveal a kind of cultural mosaic, rather than the replacement of one lithic technology by another. We believe the new round of excavations and research in the SDG site will provide more evidence for answering key questions about the process of the Middle–Upper Paleolithic transition in North China. [ABSTRACT FROM AUTHOR]

Highlights: [•] Dynamic load economic dispatch with wind power (DLEDW) model is established. [•] Markov chains combined with scenario analysis method are used to predict wind power. [•] Chance constrained technique is used to simulate the impacts of wind forecast error. [•] Improved artificial physical optimization algorithm is proposed to solve DLEDW. [•] Heuristic search strategies are applied to handle the constraints of DLEDW. [Copyright &y& Elsevier]

Highlights: [•] Second-order cone programming (SOCP) is proposed to solve unit commitment. [•] Unit commitment is transformed to SOCP model with tight lower bounds. [•] Conic quadratic constraints representation of UC problem is suggested. [•] Effectiveness of the proposed method is verified with different test systems. [•] SOCP is expanded to solve large-scale mixed integer nonlinear optimization problem. [ABSTRACT FROM AUTHOR]

Substantial quantities of sediments are known to enter the deep lithosphere at subduction zones, but the extent to which sediments melt and the process involved in sediment contribution to the deep lithosphere are inadequately understood. Vigorous debate continues about whether the subducted sediment component is terrigenous or pelagic and transported as a hydrous melt, an aqueous fluid, or bulk sediment. In this contribution, we conduct an integrated study on a variety of deep-seated xenoliths in the Neogene Hannuoba basalts from the northern margin of the North China Craton. Among these xenoliths, clinopyroxenite xenoliths are compositionally and isotopically distinct. Mineral chemistry shows that the clinopyroxenite xenoliths come from a depth near the MOHO, rather than from the mantle as suggested previously. The clinopyroxenite xenoliths have extremely evolved Sr-Nd-Hf isotopic compositions and are interpreted to have a late Archean protolith age. The extremely low contents of Cr and Ni for the clinopyroxenite xenoliths preclude a magmatic origin. Instead, a metasomatic origin is suggested, which is strongly supported for the clinopyroxenites by the occurrence of hydrous minerals and high contents of large-ion-lithophile elements (K, Rb, Ba, Th and Sr) and light rare earth elements, as well as elevated δ18O (9.9–11.3‰) and light δ26Mg (−1.04‰ to −1.42‰) isotopic compositions. Furthermore, their high high-field-strength element (Nb, Ta, Zr and Hf) contents indicate that the metasomatic agent is a hydrous melt, rather than an aqueous fluid. The metasomatic melts are considered to be derived from a mixed source of sedimentary carbonates and ancient, felsic continental materials. A combination of zircon ages and oxygen isotope data for the clinopyroxenite xenoliths further restricts the timing of metasomatism to the late Paleozoic. Considering the regional tectonic setting, the sediments most likely came from the subducted Paleo-Asian oceanic slab. Thus the Hannuoba clinopyroxenite xenoliths provide direct evidence for melting of the subducted Paleo-Asian oceanic slab sediment and its interaction with the deep lithosphere. The data show that melting of subducted sediments can take place at a much shallower depth than commonly thought and place an independent constraint on future models of slab geotherms. [ABSTRACT FROM AUTHOR]

• Cat (Felis catus) BAFF and its receptor BAFF-R were cloned. • csBAFF, EGFP/csBAFF, and cBAFF-R were expressed in E. coli and purified using a Ni2+ column. • csBAFF cellular localization was assessed. • csBAFF promoted B-cell survival in vitro. • Ligand–receptor interaction was assessed. B-cell survival depends on signals induced by binding of B-cell activating factor (BAFF) to its receptor (BAFF-R). In this study, the full-length cDNAs of cat BAFF (cBAFF) and BAFF-R (cBAFF-R) were amplified from the spleen by reverse transcription PCR. The open reading frame of cBAFF cDNA encodes a protein of 285 amino acids containing a predicted transmembrane domain and a furin protease cleavage site, similar to mammalian, avian, and reptile BAFFs. The cBAFF-R gene encodes a 189 amino acid protein. Real-time quantitative PCR analyses revealed that the two genes are predominantly expressed in the spleen. csBAFF, EGFP/csBAFF, and cBAFF-R were efficiently expressed in Escherichia coli BL21 (DE3), as confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting analyses. After purification, the EGFP/csBAFF fusion protein showed a fluorescence spectrum similar to that of EGFP. Confocal laser scanning microscopy showed that EGFP/csBAFF bound to its receptor. In vitro , csBAFF promoted the survival of cat and mouse splenic B cells with/without a priming agent (Staphylococcus aureus Cowan 1, SAC) or anti-mouse IgM. Furthermore, it stimulated the survival of mouse B cells, similar to msBAFF. Recombinant cBAFF-R blocked the function of sBAFF in vitro. These findings indicate that csBAFF plays an important role in the survival of cat B cells and has functional cross reactivity between cats and other mammals, and suggest a role for the BAFF–BAFF-R system in regulating B-cell survival. Therefore, BAFF and BAFF-R show promise for enhancing the immune systems of animals. [ABSTRACT FROM AUTHOR]

Abstract B cell activating factor from the TNF family (BAFF) is implicated in not only the physiology of normal B cells, but also the pathophysiology of aggressive B cells related to malignant and autoimmune diseases. Autophagy plays a crucial role in balancing the beneficial and detrimental effects of immunity and inflammation. However, little is known about whether and how excessive BAFF mediates autophagy contributing to B-cell proliferation and survival. Here, we show that excessive human soluble BAFF (hsBAFF) inhibited autophagy with a concomitant reduction of LC3-II in normal and B-lymphoid (Raji) cells. Knockdown of LC3 not only potentiated hsBAFF inhibition of autophagy, but also attenuated hsBAFF activation of Akt/mTOR pathway, thereby diminishing hsBAFF-induced B-cell proliferation/viability. Further, we found that hsBAFF inhibition of autophagy was Akt/mTOR-dependent. This is supported by the findings that hsBAFF increased mTORC1-mediated phosphorylation of ULK1 (Ser757); Akt inhibitor X, mTORC1 inhibitor rapamycin, mTORC1/2 inhibitor PP242, expression of dominant negative Akt, or knockdown of mTOR attenuated hsBAFF-induced phosphorylation of ULK1, decrease of LC3-II level, and increase of cell proliferation/viability. Chelating intracellular free Ca2+ ([Ca2+] i) with BAPTA/AM or preventing [Ca2+] i elevation using EGTA or 2-APB profoundly blocked hsBAFF-induced activation of Akt/mTOR, phosphorylation of ULK1 and decrease of LC3-II, as well as increase of cell proliferation/viability. Similar effects were observed in the cells where CaMKII was inhibited by KN93 or knocked down by CaMKII shRNA. Collectively, these results indicate that hsBAFF inhibits autophagy promoting cell proliferation and survival through activating Ca2+-CaMKII-dependent Akt/mTOR signaling pathway in normal and neoplastic B-lymphoid cells. Our findings suggest that manipulation of intracellular Ca2+ level or CaMKII, Akt, or mTOR activity to promote autophagy may be exploited for prevention of excessive BAFF-induced aggressive B lymphocyte disorders and autoimmune diseases. Graphical abstract Unlabelled Image Highlights • hsBAFF promotes B-cell proliferation and survival by inhibiting autophagy. • Downregulation of LC3 protein level is critical for hsBAFF inhibition in B cells. • hsBAFF-activated Akt/mTOR signaling represses autophagy via phosphorylating ULK1 (Ser757) in B cells. • hsBAFF inhibits autophagy by activating Ca2+-CaMKII-dependent Akt/mTOR signaling pathway in B cells. [ABSTRACT FROM AUTHOR]

Wheat straw (WS) was thermally pretreated at 200, 300 and 400 °C in a muffle furnace, respectively, before co-gasified with Shengli lignite (SL). The results from thermogravimetric analyzer (TGA) experiments showed that interaction of solid phases between SL and the pretreated WSs occurred at the gasification stage rather than the pyrolysis stage and the synergy varied with the biomass pretreatment temperature. Co-gasification of WS-200/SL had an inhibitory effect, and there was no synergy effect during WS-300/SL co-gasification. In contrast, co-gasification of WS-400/SL had a positive effect in the gasification stage. The investigation of co-gasification in a fixed-bed reactor, however, found synergies in product gas yields and carbon conversion for both WS-200/SL and WS-400/SL. The (H 2 +CO) yield and carbon conversion increased by 3.95% and 18.32%, respectively, for co-gasification of WS-400/SL blends compared with the calculated values based on individual materials. From the results of both the TGA and fixed-bed experiments, it is concluded that 400 °C of pyrolysis temperature for WS is the best to produce hydrogen for co-gasification with SL. The maximum synergetic effect was explained by the relatively high surface area and high content of alkali metals in WS-400. [ABSTRACT FROM AUTHOR]

Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF superfamily and plays multifunctional roles in the immune system. In the present study, a homolog of TRAIL from the Mongolian horse (named ecTRAIL) was identified and characterized. The 870-bp open reading frame encodes a polypeptide of 289 amino acid residues with a predicted molecular weight of 33.47 kDa and p I of 8.47. The genomic structure of ecTRAIL shares a five-exon/six-intron arrangement similar to its orthologs. Multiple alignments show that ecTRAIL is a type II transmembrane protein with a typical transmembrane region, three conserved cysteine residues (Cys56, Cys77, Cys238) and a TNF family signature sequence ([LV]-x-[LIVM]-x(3)-G-[LIVMF]-Y-[LIVMFY](2)-x(2)-[QEKHL]-[LIVMGT]-x-[LIVMFY]). Three-dimensional structure prediction based on the same template revealed that the positional arrangement of the key amino acid residues, Cys238 and Cys230 in ecTRAIL and human TRAIL, respectively, is significantly conserved. Evolutionary analysis suggests that ecTRAIL is most closely related to its ortholog from pigs, with an identity of 83.99%. The solubilizing small ubiquitin-related modifier (SUMO) tag fused recombinant protein SUMO-ecsTRAIL was successfully expressed in E. coli and exhibited binding activity and cytotoxicity to HeLa cells in a cross-species manner in vitro . These results provide a better understanding of TRAILs in mammals and indicate that ecTRAIL may play an important role in the immune response in horses. [ABSTRACT FROM AUTHOR]

[Display omitted] Therapeutically targeting B cells has received great attention in the treatment of B-cell malignancies and autoimmune diseases. The B-cell activating factor (BAFF) is critical to the survival of normal and neoplastic B cells, and excess production of BAFF contributes to autoimmune diseases. Resveratrol, a natural polyphenolic compound, has a positive effect on the treatment of autoimmune diseases. However, how resveratrol affects BAFF-stimulated B-cell proliferation and survival is poorly understood. Here, we show that resveratrol increased autophagosome formation and ATG5/LC3-II levels and decreased p62 level, promoting autophagic flux/autophagy and thereby suppressing the basal or human soluble BAFF (hsBAFF)-stimulated proliferation and survival of normal and B-lymphoid (Raji) cells. This is supported by the findings that inhibition of autophagy with 3-methyladenine (3-MA, an inhibitor of Vps34) or ATG5 shRNA attenuates resveratrol-induced autophagy and -reduced proliferation/viability in B-cells. Inhibition of mTOR with rapamycin or knockdown of mTOR potentiated resveratrol-induced autophagy and inhibition of hsBAFF-stimulated B-cell proliferation/viability, while overexpression of wild-type mTOR conferred resistance to the actions of resveratrol. Similarly, inhibition of Akt with Akt inhibitor X or ectopic expression of dominant negative Akt reinforced resveratrol-induced autophagy and inhibition of hsBAFF-stimulated B-cell proliferation/viability, whereas expression of constitutively active Akt conferred resistance to the actions of resveratrol. Taken together, these results indicate that resveratrol induces autophagy impeding BAFF-stimulated proliferation and survival via blocking the Akt/mTOR signaling pathway in normal and neoplastic B cells. Our findings highlight that resveratrol has a great potential for prevention and treatment of excessive BAFF-elicited aggressive B-cell disorders and autoimmune diseases. [ABSTRACT FROM AUTHOR]

Few sites in China have yielded definite evidence for a complex bone technology. In this paper, we present a detailed description and an integrated techno-functional study of seven bone needles from Shuidonggou locality 12 dated to 12.2–11.0 ka BP in North China. Tool production processes conforming to the classic descriptions of formal techniques, including scraping, grinding, perforating, polishing and occasionally heat-treating were streamlined to process osseous materials at this site. In Shuidonggou locality 12, the bone needles manufactured accordingly were used to produce sophisticated and fitted clothing, which helps to counter winter subsistence risk, until they were very blunt or broken or no more appropriate for use, and then were discarded in situ. [ABSTRACT FROM AUTHOR]

The mysterious loss of five calvaria of Homo erectus and three skulls of Homo sapiens sapiens , unearthed from Zhoukoudian, a world cultural heritage site, during World War II is a great loss for paleoanthropological research. Although frontal and occipital bones of Homo erectus were unearthed in 1966, additional important human fossils cannot be found due to the fact that most of the fossil-bearing deposits were removed in early diggings at ZKD Locality 1, the so-called Peking Man Site. From 1995 to 2004, a series of in situ geological and geophysical investigations using electromagnetic, electric panel, magnetic, microgravity, electric tomograms, surface seismic waves, ground penetrating radar, and test drilling methods, were conducted on Dragon Bone Hill, where the Peking Man Site is located. Here, we report that some geophysical anomalies have been clearly detected and there are still significant deposits suitable for archaeological excavations in hidden cavities in the hill, which demonstrate how some civil engineering methods can be employed for archaeological purposes. [ABSTRACT FROM AUTHOR]

Protostane triterpenes from Alisma orientale (Sam.) Juz. have exhibited distinct pharmacological properties that are currently in high demand. 3-Hydroxy-3-methylglutaryl-CoA reductase (HMGR) is considered the first rate-limiting enzyme in isoprenoid biosynthesis via the mevalonic acid (MVA) pathway. In this study, we cloned a full-length cDNA of A. orientale (Sam.) Juz. HMGR (AoHMGR; 2252 bp; GenBank accession no. KP342318) with an open reading frame (ORF) of 1809 bp. The deduced protein sequence contained four conserved motifs and exhibited homology with HMGR proteins from other plants. We next expressed the cloned gene in Escherichia coli BL21 (Rosetta) cells, collected the expressed products, and incubated those with 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) to determine enzymatic activity. GC/MS analysis revealed that the products were able to catalyze HMG-CoA and NADPH to form MVA. The purified protein was used to immunize New Zealand rabbits and prepare an antibody against AoHMGR. Western blot results demonstrated that the antibodies specifically recognized AoHMGR protein in A. orientale (Sam.) Juz. We then established a rapid test to detect AoHMGR protein in the plant, and found the tuber to be the most AoHMGR protein-abundant organ in A. orientale (Sam.) Juz. Furthermore, we detected the expression level of AoHMGR and contents of the main active component, Alisol B 23-acetate, at different growth phases of A. orientale (Sam.) Juz. A significant positive correlation was identified, indicating that AoHMGR represents a key enzyme in the synthetic pathway of protostane triterpenes. [ABSTRACT FROM AUTHOR]

Abstract: B cell activating factor (BAFF) and its receptors were regarded as elements of the immune system, regulating the fate of B cell. In recent years, these molecules were identified in a number of normal and pathological tissues, expanding their potential functions beyond the immune system. In this study, on the basis of molecular clone and prokaryotic expression of equine BAFF, we reported that equine adipose-derived stem cell (ASC) expressed BAFF and its receptors, which exhibited the increased expression during ASC adipogenic differentiation in vitro. Moreover, with the addition of recombinant protein His6-sBAFF, an increased differentiation of equine ASC towards adipocyte was detected. These results suggested that BAFF and its receptors might be associated with the differentiation process of ASC towards adipocyte in horse. [Copyright &y& Elsevier]

Abstract: B-cell activating factor (BAFF) is involved in not only the physiology of normal B cells, but also the pathophysiology of aggressive B cells related to malignant and autoimmune diseases. However, how excessive BAFF promotes aggressive B-cell proliferation and survival is not well understood. Here we show that excessive human soluble BAFF (hsBAFF) enhanced cell proliferation and survival in normal and B-lymphoid (Raji) cells, which was associated with suppression of PP2A, resulting in activation of Erk1/2. This is supported by the findings that pretreatment with U0126 or PD98059, expression of dominant negative MKK1, or overexpression of PP2A prevented hsBAFF-induced activation of Erk1/2 and cell proliferation/viability in the cells. It appears that hsBAFF-mediated PP2A-Erk1/2 pathway and B-cell proliferation/viability was Ca2+-dependent, as pretreatment with BAPTA/AM, EGTA or 2-APB significantly attenuated these events. Furthermore, we found that inhibiting CaMKII with KN93 or silencing CaMKII also attenuated hsBAFF-mediated PP2A-Erk1/2 signaling and B-cell proliferation/viability. The results indicate that BAFF activates Erk1/2, in part through Ca2+-CaMKII-dependent inhibition of PP2A, increasing cell proliferation/viability in normal and neoplastic B-lymphoid cells. Our data suggest that inhibitors of CaMKII and Erk1/2, activator of PP2A or manipulation of intracellular Ca2+ may be exploited for prevention of excessive BAFF-induced aggressive B-cell malignancies and autoimmune diseases. [Copyright &y& Elsevier]

Abstract: B-cell survival depends on signals induced by B-cell activating factor (BAFF) that binds to the BAFF receptor (BAFF-R). Herein, a BAFF-R homolog was identified in a horse (Equus caballus). The horse BAFF-R gene, located on chromosome 28, spans 1444 base pairs and encodes a 183-amino acid protein. The protein is structurally conserved, in which the DxL motif plays an important role in binding to BAFF. Furthermore, the horse BAFF-R extracellular domain was expressed and purified, which specifically bound to His6-sBAFF and had the capability of blocking the function of His6-sBAFF in vitro. Finally, evolutionary analyses indicated that some codon sites of BAFF-R evolve with positive selection and that the genetic relationship among a horse, Chiroptera, and Caniformia are the closest. [Copyright &y& Elsevier]

Highlights: [•] An GILT homologue has been cloned and identified from Mefugu. [•] The expression pattern of GILT gene is investigated in various tissues. [•] The mGILT expression is obviously up-regulated in spleen and kidney after immunization with LPS. [•] We also describe the expression and bioactivity analysis of GILT protein. [ABSTRACT FROM AUTHOR]

Abstract: LIGHT (lymphotoxin-related inducible ligand that competes with herpes simplex virus (HSV) glycoprotein D for herpesvirus entry mediator on T cells) is a member of the tumor necrosis factor (TNF) ligand superfamily, which plays important roles in inflammatory and immune responses. In the present study, the cDNAs of guinea pig (Cavia porcellus) LIGHT (designated as gpLIGHT) and its receptor herpes virus entry mediator (designated as gpHVEM) were amplified from spleen by reverse transcription polymerase chain reaction (RT-PCR). The ORFs of gpLIGHT and gpHVEM cover 726 and 861bp, encoding predicted proteins with 241 and 286 aas, respectively. The three-dimensional (3D) structure, phylogenetic relationships, and characterization of both genes were also analyzed. We also generated a 3D model to verify interaction between the two proteins. Real-time quantitative PCR (qPCR) analysis revealed that both LIGHT and HVEM are constitutively expressed in guinea pig various tissues. A fusion protein SUMO (Small Ubiquitin-like Modifier)-gpsLIGHT (the soluble mature part of gpLIGHT) was efficiently expressed in Escherichia coli BL21 (DE3) and purified using metal chelate affinity chromatography (Ni-NTA). Laser scanning confocal microscopy (LSCM) showed that gpsLIGHT can bind its receptors on T cells. The LIGHT-HVEM signaling pathway plays an important role in the immune system, and our results might provide a platform for further research into the effects of LIGHT and HVEM. [Copyright &y& Elsevier]

Abstract: Oryctolagus cuniculus is an important animal model which is widely used for vaccine development, and investigations of human infectious and autoimmune diseases. TNFSF14 is one of the most important TNF superfamily cytokines playing roles in lymphocyte homeostasis by enhancing effector cell activation and survival, or by cellular elimination through apoptosis. For these applications, we cloned full-length cDNA encoding TNFSF14 and its receptor LTBR from O. cuniculus. Then we analyzed the genes structure, function, evolutionary relationships, and predicted three-dimensional structure of proteins and proteins’ interaction model. Recombinant O. cuniculus soluble TNFSF14 (OcsTNFSF14) and extracellular LTBR (OceLTBR) proteins were efficiently produced in Escherichia coli BL21 (DE3) and purified by Ni-NTA affinity chromatography, then confirmed by SDS–PAGE and Western Blotting analysis. Confocal laser microscopy assays revealed that OcsTNFSF14 and OceLTBR could bind splenic T cells in vitro, suggesting a series of signal pathway activated inside cells in response to OcsTNFSF14 and OceLTBR. The obtained results will lay the foundation for future study of human TNFSF14/LTBR signal pathway and relative diseases in O. cuniculus model. [Copyright &y& Elsevier]

Abstract: B cell activating factor (BAFF), a member of the tumor necrosis factor family, is critical to B cell survival, proliferation, maturation, and immunoglobulin secretion and to T cell activation. In the present study, the full-length cDNA of BAFF from the South African clawed frog (Xenopus laevis, designated xlBAFF) was cloned using rapid amplification of cDNA ends (RACE) techniques and RT-PCR. The full-length cDNA of xlBAFF consists of 1204 bases including an open reading frame (ORF) of 801 nucleotides that are translated into a predicted 266 amino acid protein. Sequence comparison indicated that the amino acids of xlBAFF possessed the TNF signature, including a transmembrane domain, a putative furin protease cleavage site and three cysteine residues. The predicted three-dimensional (3D) structure of the xlBAFF monomer revealed that it was very similar to its counterparts. Real-time quantitative PCR analysis revealed that xlBAFF could be detected in various tissues and predominantly expressed in the spleen and other lymphoid tissue. The soluble xlBAFF had been cloned into a pET28a vector to express the recombinant protein. The His6-xlBAFF was efficiently expressed in Escherichia coli. BL21 (DE3) and its expressions were confirmed by SDS-PAGE and Western blotting analysis. After purification, laser scanning confocal microscopy analysis showed that xlBAFF could bind to its receptors on B cells. CCK-8 assays revealed that xlBAFF is not only able to promote survival/proliferation of South African clawed frog lymphocytes but also able to stimulate survival/proliferation of mouse B cells. These results will allow for further investigation the use of X. laevis as an in vivo model for related studies. [Copyright &y& Elsevier]

Abstract: B cell activating factor (BAFF), a member of the TNF family, is a critical cytokine for the survival, proliferation, maturation, and differentiation of B cells. In the present study, Père David''s deer BAFF (miBAFF) was amplified from Elaphurus davidianus using RT-PCR. This is the first BAFF cloned from a member of Cervidae family. The open reading frame (ORF) of the miBAFF cDNA consists of 843 bases that encode a 280-amino acid protein bearing typical TNF homology domain. Sequence alignment shows that miBAFF shares 39.3%–97% sequence homology with the BAFF sequences of other mammals. Comparative protein modeling predicted that the 3D structure of the soluble mature portion of miBAFF (misBAFF) is very similar to that of human BAFF (hsBAFF). Recombinant misBAFF fused to a SUMO-tag was efficiently expressed in Escherichia coli BL21 (DE3) cells. The protein molecular weight of ~36KDa was determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. In vitro, purified misBAFF was shown to promote the survival and proliferation of Père David''s deer peripheral blood lymphocytes and mouse B cells. These results indicate that miBAFF plays an important role in the survival/proliferation of mouse B cells and, shows highly conserved evolutionarily, leading to functional cross-reactivity that exists between mouse and Père David''s deer BAFF. [Copyright &y& Elsevier]

Abstract: Here we describe the identification of a Danio rerio homologue of a proliferation-inducing ligand (APRIL) of the TNF family (designated zAPRIL). Sequence analysis showed that the open reading frame of zAPRIL consists of 600 bases encoding a protein of 199-amino acids. Recombinant soluble APRIL (zsAPRIL) was constructed consisting of fluorescence-enhanced green fluorescent protein (EGFP) and cloned into a pET28a vector. SDS–PAGE and western blotting analysis indicated a high-level expression of soluble EGFP/zsAPRIL protein in Escherichia coli BL21 (DE3). Observation by confocal microscopy demonstrated that EGFP/zsAPRIL could successfully bind to the surface receptors of zebrafish lymphocytes. In vitro survival analysis revealed that purified EGFP/zsAPRIL was able to promote the survival of zebrafish lymphocytes in a dose-dependent manner. The biological role of APRIL does not seem to be restricted to proliferation induction. Zebrafish may could served as a model organism for further study of APRIL. [Copyright &y& Elsevier]