AZGP1 as a potential biomarker of IgA vasculitis with nephritis in a children­based urinary proteomics study by diaPASEF.

The present study aimed to screen for potential biomarkers in the urine of immunoglobulin A vasculitis with nephritis (IgAVN) using parallel accumulation­serial fragmentation combined with data­independent acquisition (diaPASEF) proteomic approach. Urine proteomes of eight children with IgAVN and eight healthy children were identified by diaPASEF and all differential proteins were analyzed by Gene Ontology and Kyoto Encyclopedia of Gene and Genome. Then, the specific biomarkers of urine samples from 10 children with IgAVN, 10 children with IgAV and 10 healthy children were verified by ELISA. The present study screened 254 differential proteins from the experiment, including 190 upregulated proteins and 64 downregulated proteins. The results of the ELISA showed that the concentration of urinary zinc­alpha­2­glycoprotein (AZGP1) in children with IgAVN was significantly higher compared with that in children with IgAV and healthy children. The present study provided the potential clinical application of AZGP1 as a helpful biomarker and a potential indicator for early diagnosis of the occurrence of IgAVN.

Developing a dynamic magnetic flux template to guide 1D nanomaterial growth.

A dynamic magnetic flux template (DMT) has been developed for preparing 1D nanomaterials efficiently. It can be quickly established and revoked without introducing any pollution. The DMT behaves like a template for guiding the orientation, bearing a 1D structure, activating the nucleation, and providing a driving force for 1D nanomaterial growth.

Chemokine (C-C motif) receptor 2 is associated with the pathological grade and inflammatory response in IgAN children.

BACKGROUND: Chemokine (C-C motif) receptor 2 (CCR2) is involved in important physiological and pathological processes, such as inflammation and autoimmune diseases. Abnormal immune and inflammatory responses play a critical role in the development and progression of IgA nephritis (IgAN). However, the role of CCR2 in IgAN is unknown. METHODS: Fifteen IgAN children who were diagnosed by kidney biopsy provided kidney biopsy tissue, blood and urine samples, and age-matched healthy control subjects (blood donators n = 12; tissue donators n = 8) were included. Immunohistochemical analysis was used to detect the expression of CCR2, MCP-1, IL-6, IL-17, and TNF-α in the kidney tissues. Relative optical density (OD) was calculated by Image J software, and the correlation between CCR2 expression and pathological grade in IgAN children was analyzed. RESULTS: The expression of CCR2 significantly increased in mesangial cells of children with IgAN compared to that in control group (P < 0.001), especially in IgAN patients with Lee's grade III to IV (P < 0.001). Interestingly, CCR2 expression was positively correlated with Lee's grade (r = 0.9152, P = 0.0001) in IgAN children. The expression levels of inflammatory factors were markedly increased in IgAN children, and importantly CCR2 expression was positively correlated with it's expression level. CONCLUSIONS: The results suggest that CCR2 signaling might be involved in pathological process and inflammatory responses of children IgAN, and could potentially be an intervention target in children IgAN.

[Electroacupuncture at "Zusanli"(ST36) protects intestinal mucosal immune barrier by suppre-ssing apoptosis of intestinal lymphocytes and regulating expression of Bcl-2 and Bax in sepsis rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zusanli" (ST36) on apoptosis of intestinal T lymphocytes, translocation of intestinal bacteria and expression of intestinal Bcl-2 and Bax proteins and intestinal mucosal immune barrier in sepsis rats, so as to explore its underlying mechanism in relieving sepsis. METHODS: SD rats were randomly divided into sham operation (n=6), model (n=15), non-meridian and non-acupoint (non-acupoint, n=15) and acupoint EA(n=15) groups by using random number table method. The sepsis model was established by using cecal ligation and perforation(CLP) method. EA (2 Hz, 2 mA) was applied to bilateral ST36 or non-acupoint for 30 min one hour after modeling, once every day for 3 days. The rats' general conditions and fatality rate in 3 days after modeling were recorded. The liver, spleen and mesenteric lymph nodes were taken for bacterial culture to detect the translocation rate of intestinal bacteria. The small intestinal tissue was taken for observing histopathological changes (Chiu's score: 0-5 points) after HE staining, and for determining the expression levels of Bcl-2 and Bax proteins using Western blot. The intestinal mucosa was sampled for detecting the apop-tosis (apoptotic index) of lymphocytes by using terminal deoxynucleoitidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling (TUNEL) assay, and the counts of CD4+ and CD8+T cells using flow cytometry. The contents of IL-4 in the small intestine and that of secretory IgA (sIgA) in the small intestinal mucus were determined by using ELISA. RESULTS: After modeling, of the 15 rats in each of the 3 groups, 7, 7 and 2 in the model, non-acupoint and EA groups were dead in the first 3 days, with the fatality rate being 46.67% (7/15), 46.67% (7/15) and 13.33% (2/15), respectively (being obviously lower in the EA group than in the former two groups, P<0.05). Compared with the sham operation group, the incidence of intestinal bacterial translocation, apoptotic index, Chiu's score, and Bax expression were significantly increased (P<0.05), and the percentages of CD4+ and CD8+T cells, IL-4 and sIgA contents and Bcl-2 expression considerably decreased (P<0.05) in the model group. In comparison with the model group, modeling-induced increase of incidence of bacterial translocation, apoptotic index and Bax expression, and decrease of percentages of CD4+ and CD8+T cells, IL-4 and sIgA contents and Bcl-2 expression were reversed (P<0.05) in the EA group. CONCLUSION: EA at ST36 can reduce death rate and intestinal bacteria translocation incidence in sepsis rats, which may be related to its functions in regulating the expression of intestinal Bcl-2 and Bax proteins and inhibiting the apoptosis of intestinal mucosal T lymphocytes, thereby protecting the immune barrier function of intestinal mucosa to reduce the intestinal permeability.

Preliminary Exploration of Economic Polypropylene-Fiber-Reinforced ECC with Superfine River Sand (SSPP-ECC) Applied to a Bridge Pavement Leveling Overlay.

In response to the current common disease of concrete leveling overlays of bridge pavement in China, the feasibility of using an economic SSPP-ECC with local waste superfine sand as an alternative material for a leveling overlay was proposed in this study. To evaluate the interface bonding property in the girder between the SSPP-ECC and concrete, a slant shear test and split tensile test were designed to study the interfacial shear and tensile properties of the ordinary concrete/ordinary concrete (OC/OC) and ordinary concrete/SSPP-ECC (OC/ECC), where the results showed that SSPP-ECC could significantly improve the interface shear stress and split tensile strength compared to ordinary concrete. Furthermore, the damage status of OC/ECC no longer involved fracturing along the interface; instead, each of the two substrates was partially destroyed, which revealed that OC/ECC had a high bonding effect. Moreover, a restrained shrinkage test was carried out to evaluate the shrinkage property of SSPP-ECC, where the result showed that the shrinkage strain of SSPP-ECC was slightly lower than concrete, where the average cracking time for SSPP-ECC was far longer than for ordinary concrete under the same ambient drying conditions; furthermore, the stress rate for SSPP-ECC revealed that it was a low-cracking-risk material. Meanwhile, the crack width of SSPP-ECC was only 0.1 mm after 35 d, which showed that SSPP-ECC had a more substantial crack width control capacity relative to concrete. The test results initially verified the feasibility and great potential of economic SSPP-ECC applied in a bridge pavement leveling overlay.

Extensive Diversity and Prevalent Fluconazole Resistance among Environmental Yeasts from Tropical China.

Yeasts play important roles in both the environment and in human welfare. While some environmental yeasts positively contribute to nutrient cycling and food production, a significant number of yeast species are opportunistic human pathogens, including several that are tolerant/resistant to commonly used antifungal drugs. At present, most of our understanding of environmental yeasts has come from a few terrestrial environments in selected geographic regions. Relatively little is known about yeast diversity in tropical environments and their potential impacts on human health. Here, we characterize culturable yeasts in 968 environmental samples from eight regions in tropical China. Among the 516 soil, 273 freshwater, and 179 seawater samples, 71.5%, 85.7%, and 43.6% contained yeasts, respectively. A total of 984 yeast isolates were analyzed for their DNA barcode sequences and their susceptibilities to fluconazole. DNA sequence comparisons revealed that the 984 yeast isolates likely belonged to 144 species, including 106 known species and 38 putative novel species. About 38% of the 984 isolates belonged to known human pathogens and the most common species was Candida tropicalis, accounting for 21% (207/984) of all isolates. Further analyses based on multi-locus sequence typing revealed that some of these environmental C. tropicalis shared identical genotypes with clinical isolates previously reported from tropical China and elsewhere. Importantly, 374 of the 984 (38%) yeast isolates showed intermediate susceptibility or resistance to fluconazole. Our results suggest that these environmental yeasts could have significant negative impacts on human health.

Establishment of Complex Modulus Master Curves Based on Generalized Sigmoidal Model for Freeze-Thaw Resistance Evaluation of Basalt Fiber-Modified Asphalt Mixtures.

This study aims to study the freeze-thaw (F-T) resistance of asphalt mixture incorporating styrene-butadiene-styrene (SBS) polymer and basalt fiber by using the established complex master curves of the generalized Sigmoidal model. Asphalt mixture samples incorporating styrene-butadiene-styrene (SBS) polymer and basalt fiber were manufactured following the Superpave gyratory compaction (SGC) method and coring as well as sawing. After 0-21 F-T cycles processing, a complex modulus test asphalt mixture specimen was performed to evaluate the influence of the F-T cycle. Besides, according to the time-temperature superposition principle, the master curves of a complex modulus were constructed to reflect the dynamic mechanical response in an extended range of reduced frequency at an arbitrary temperature. The results indicated that the elastic and viscous portions of asphalt mixture incorporating SBS and basalt fiber have decreased overall. It could be observed from the dynamic modulus ratio that the dynamic modulus ratios of specimens were more affected by the F-T cycle at low frequency or high temperature. Thus, in the process of asphalt pavement design and maintenance, attention should be paid to seasonal frozen asphalt pavement under low frequency and high temperature.

Master Curve Establishment and Complex Modulus Evaluation of SBS-Modified Asphalt Mixture Reinforced with Basalt Fiber Based on Generalized Sigmoidal Model.

Basalt fiber has been proved to be a good modified material for asphalt mixture. The performance of basalt fiber modified asphalt mixture has been widely investigated by extensive researches. However, most studies focused on ordinary static load tests, and less attention was paid to the dynamic mechanical response of asphalt mixture incorporating with basalt fiber. This paper aims to establish the master curve of complex modulus of asphalt mixture incorporating of styrene-butadiene-styrene (SBS) polymer and basalt fiber using the generalized Sigmoidal model. Both loading frequency and temperature were investigated for dynamic mechanical response of asphalt mixture with basalt fiber. In addition, based on the time-temperature superposition principle, the master curves of complex modulus were constructed to reflect the dynamic mechanical response at an extended reduced frequency range at an arbitrary temperature. Results indicated that the generalized Sigmoidal model in this paper could better reflect the dynamic mechanical response accurately with correlation coefficients above 0.97, which is utilized to predict the dynamic mechanical performances accurately. Simultaneously, the modulus values exhibit an increasing trend with loading frequency and decrease versus temperature. However, the phase angle values showed different trends with frequency and temperature.

Preliminary Analysis of the Ductility and Crack-Control Ability of Engineered Cementitious Composite with Superfine Sand and Polypropylene Fiber (SSPP-ECC).

Engineered cementitious composite (ECC) is a potential cement-based material with the abilities of large deformation and crack width control. However, ECC is difficult to popularize in many developing countries because the costs of silica sand and polyvinyl alcohol (PVA) fiber with a surface coating are too high for practical engineering. Therefore, we proposed an economical ECC with superfine river sand and polypropylene (PP) fiber (SSPP-ECC) to replace PVA fiber and silica sand. The SSPP-ECC proposed in this paper is a sustainable material using local material ingredients, which has considerable adaptability for large-scale engineering applications. The 16 groups of specimens were prepared through a factorial design method, curing for four-point bending tests. The bending strength, deflection, flexural modulus of elasticity, and crack width were measured and calculated during the test. The factor analysis of the test results shows that the contents of fiber and fly ash had significant effects on the ductility of SSPP-ECC with an extra combined effect at the same time, and a response surface model with high accuracy was fitted to predict the yield length of SSPP-ECC. The ductility of SSPP-ECC was positively related to its crack-control ability and it was shown that the crack width of SSPP-ECC increased significantly with a high content of superfine sand. This paper proposed a reasonable way to utilize superfine sand and provided the mix proportion of SSPP-ECC with characteristics of deformation hardening and multi-cracking, which may cater to the demands of many concrete components on ductility and crack resistance.

Reducing Energy Penalty of CO2 Capture Using Fe Promoted SO42-/ZrO2/MCM-41 Catalyst.

The high energy consumption of CO2-loaded solvent regeneration is the biggest impediment for the real application of the amine-based CO2 capture process. To lower the energy requirement, three Fe promoted SO42-/ZrO2 supported on MCM-41 (SZMF) catalysts with different iron oxide content (5%, 10%, and 15%) were synthesized and applied for the rich monoethanolamine solution regeneration process at 98 °C. Results reveal that the use of SZMF hugely enhanced the CO2 desorption performances (i.e., desorption factor) by 260-388% and reduced the heat duty by about 28-40%, which is better than most of the reported catalysts for this purpose. The eminent catalytic activities of SZMF are related to their enhanced ratio of Brønsted to Lewis acid sites, weak acid sites, basic sites, and high dispersed Fe3+ species. Meanwhile, the addition of SZMF for CO2 desorption shows a promotional effect on its CO2 absorption performance, and SZMF presents an excellent cyclic stability. A possible mechanism is suggested for the SZMF catalyzed CO2 desorption process. Results of this work may provide direction for future research and rational design of more efficient catalysts for this potential catalyst-aided CO2 desorption technology.

Gene expression in chronic granulomatous disease and interferon-γ receptor-deficient cells treated in vitro with interferon-γ.

Interferon-γ (IFN-γ) plays an important role in innate and adaptive immunity against intracellular infections and is used clinically for the prevention and control of infections in chronic granulomatous disease (CGD) and inborn defects in the IFN-γ/interleukin (IL)-12 axis. Using transcriptome profiling (RNA-seq), we sought to identify differentially expressed genes, transcripts and exons in Epstein-Barr virus-transformed B lymphocytes (B-EBV) cells from CGD patients, IFN-γ receptor deficiency patients, and normal controls, treated in vitro with IFN-γ for 48 hours. Our results show that IFN-γ increased the expression of a diverse array of genes related to different cellular programs. In cells from normal controls and CGD patients, IFN-γ-induced expression of genes relevant to oxidative killing, nitric oxide synthase pathway, proteasome-mediated degradation, antigen presentation, chemoattraction, and cell adhesion. IFN-γ also upregulated genes involved in diverse stages of messenger RNA (mRNA) processing including pre-mRNA splicing, as well as others implicated in the folding, transport, and assembly of proteins. In particular, differential exon expression of WARS (encoding tryptophanyl-transfer RNA synthetase, which has an essential function in protein synthesis) induced by IFN-γ in normal and CGD cells suggests that this gene may have an important contribution to the benefits of IFN-γ treatment for CGD. Upregulation of mRNA and protein processing related genes in CGD and IFNRD cells could mediate some of the effects of IFN-γ treatment. These data support the concept that IFN-γ treatment may contribute to increased immune responses against pathogens through regulation of genes important for mRNA and protein processing.

Dual-cutoff of sFlt-1/PlGF ratio in the stratification of preeclampsia: a systematic review and meta-analysis.

PURPOSE: To systematically review the approach of using two independent sFlt-1/PlGF cutoffs that has better sensitivity (cutoff-sen) and specificity (cutoff-spe) separately for risk stratification in the detection of preeclampsia. METHODS: PubMed and Embase databases and reference lists were searched up to June 2016. Inclusion criteria were blood samples for sFlt-1/PlGF with separate cutoffs (cutoff-sen and cutoff-spe) provided. Six relevant studies were identified. Pooling of results was done based on three studies and a systematic review was performed based on all six. RESULTS: The strategy of using a cutoff of ≤33 and ≥85 for early onset preeclampsia, and ≤33 and ≥110 for the late onset preeclampsia was proposed and examined. The pooled sensitivity for cutoff-sen was: 95.3% (90.6-98.1%) and 88.6% (82.9-92.9%) for early and late onset preeclampsia, respectively. The pooled specificity for cutoff-spe was: 97.6% (95.2-98.9%) and 94.2% (91.4-96.3%) for early and late onset preeclampsia respectively. The pooled estimation of the early onset pre-eclamptic pregnancies and control normal pregnancies classified in the equivocal zone was 4.9% (2.0-8.8%) and 32.4% (25.7-39.5%), respectively, and 26.8% (10.3-47.6%) and 8.7% (3.0-17.6%) for late onset patients. CONCLUSION: The new dual-cutoff diagnostic system optimizes the predictive performance of the single cutoff system. Further studies are required to assess the performance of this system and to define the approach and frequency at which subjects in the equivocal zone should be screened.

Interactions between the Microtubule Binding Protein EB1 and F-Actin.

Many cellular processes including cell division and cell migration require coordination between the actin and microtubule (MT) cytoskeletons. This coordination is as-yet poorly understood, but proteins such as formins and IQGAP1 are known to be involved. We show that the MT binding protein EB1 (end-binding protein 1), a key regulator of MT dynamics, can bind directly to filamentous actin (F-actin) F-actin. We determined that the EB1:F-actin interaction is salt sensitive and weak under physiological salt concentrations but might be relevant in contexts where the local concentration of actin is high. Using bioinformatics and mutagenesis, we found that the EB1:F-actin binding site partially overlaps the well-characterized EB1:MT binding interface. Congruently, competition experiments indicate that EB1 can bind to F-actin or MTs but not both simultaneously. These observations suggest that EB1:F-actin interactions may negatively regulate EB1:MT interactions, and we speculate that this interaction may assist cells in differentially regulating MT stability in the actin-rich cortex as opposed to the cell interior.

Regulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF-κB Binding Site.

The human CYBB gene encodes the gp91-phox component of the phagocyte oxidase enzyme complex, which is responsible for generating superoxide and other downstream reactive oxygen species essential to microbial killing. In the present study, we have identified by sequence analysis a putative NF-κB binding site in a DNase I hypersensitive site, termed HS-II, located in the distant 5' flanking region of the CYBB gene. Electrophoretic mobility assays showed binding of the sequence element by recombinant NF-κB protein p50 and by proteins in nuclear extract from the HL-60 myeloid leukemia cell line corresponding to p50 and to p50/p65 heterodimers. Chromatin immunoprecipitation demonstrated NF-κB binding to the site in intact HL-60 cells. Chromosome conformation capture (3C) assays demonstrated physical interaction between the NF-κB binding site and the CYBB promoter region. Inhibition of NF-κB activity by salicylate reduced CYBB expression in peripheral blood neutrophils and differentiated U937 monocytic leukemia cells. U937 cells transfected with a mutant inhibitor of κB "super-repressor" showed markedly diminished CYBB expression. Luciferase reporter analysis of the NF-κB site linked to the CYBB 5' flanking promoter region revealed enhanced expression, augmented by treatment with interferon-γ. These studies indicate a role for this distant, 15 kb upstream, binding site in NF-κB regulation of the CYBB gene, an essential component of phagocyte-mediated host defense.

Dicer cooperates with p53 to suppress DNA damage and skin carcinogenesis in mice.

Dicer is required for the maturation of microRNA, and loss of Dicer and miRNA processing has been found to alter numerous biological events during embryogenesis, including the development of mammalian skin and hair. We have previously examined the role of miRNA biogenesis in mouse embryonic fibroblasts and found that deletion of Dicer induces cell senescence regulated, in part, by the p53 tumor suppressor. Although Dicer and miRNA molecules are thought to have either oncogenic or tumor suppressing roles in various types of cancer, a role for Dicer and miRNAs in skin carcinogenesis has not been established. Here we show that perinatal ablation of Dicer in the skin of mice leads to loss of fur in adult mice, increased epidermal cell proliferation and apoptosis, and the accumulation of widespread DNA damage in epidermal cells. Co-ablation of Dicer and p53 did not alter the timing or extent of fur loss, but greatly reduced survival of Dicer-skin ablated mice, as these mice developed multiple and highly aggressive skin carcinomas. Our results describe a new mouse model for spontaneous basal and squamous cell tumorigenesis. Furthermore, our findings reveal that loss of Dicer in the epidermis induces extensive DNA damage, activation of the DNA damage response and p53-dependent apoptosis, and that Dicer and p53 cooperate to suppress mammalian skin carcinogenesis.

Investigation of monoclonal antibody fragmentation artifacts in non-reducing SDS-PAGE.

Fragmentation of monoclonal antibodies has been routinely observed in non-reducing SDS-PAGE, mainly due to disulfide-bond scrambling catalyzed by free sulfhydryl groups, resulting in a method induced artifact. To minimize this artifact, alkylating agents like iodoacetamide (IAM) and N-ethylmaleimide (NEM) were commonly included in SDS sample buffer to block free sulfhydryls. However, the selection of agents and the applied concentrations differ from study to study. In addition, there is no direct comparison of these agents thus far, resulting in difficulties in selecting the suitable agent. To address these questions, we have tested the activities of IAM and NEM in inhibiting the fragment-band artifact of IgG4 monoclonal antibodies. Our data suggest that the inhibition activity of both agents is concentration dependent. Interestingly, 5mM NEM can achieve the same inhibition effect as 40 mM IAM. In addition, NEM still retained strong activity after prolonged sample heating, whereas IAM lost most of its activity. Overall, NEM appears to have a better inhibition effect than IAM on all tested IgG4 proteins, either with SDS-PAGE or CE-SDS methods. These observations demonstrate that NEM has stronger fragmentation inhibition activity than IAM, and thus is a more suitable alkylating agent for both SDS-PAGE and CE-SDS method to reduce this fragmentation artifact.

Syntheses and biological evaluation of ring-C modified colchicine analogs.

Ring-C modified alkaloids were synthesized from colchicine using iminonitroso Diels-Alder reactions in a highly regio- and stereoselective fashion. Several analogs exhibited cytotoxic activity similar to that of colchicine itself against PC-3 and MCF-7 cancer cell lines, by serving as prodrugs of colchicine through retro Diels-Alder reactions under the assayed conditions. In vitro microtubule polymerization assays indicated that these modifications affected their interaction with tubulin.

Cyclic neutropenia and severe congenital neutropenia in patients with a shared ELANE mutation and paternal haplotype: evidence for phenotype determination by modifying genes.

BACKGROUND: Cyclic neutropenia (CN) and severe congenital neutropenia (SCN) are disorders of neutrophil production that differ markedly in disease severity. Mutations of the ELANE gene (the symbol recently replacing ELA2) are considered largely responsible for most cases of CN and SCN, but specific mutations are typically associated with one or the other. PROCEDURE: We performed ELANE genotyping on all individuals and paternal sperm in an SCN kindred with eight SCN progeny of a sperm donor and six different mothers. RESULTS: One patient with CN had the same S97L ELANE mutation as seven patients with the SCN phenotype. The mutant allele was detected in the donor's spermatozoa, representing 18% of the ELANE gene pool, but not in DNA from his lymphocytes, neutrophils, or buccal mucosa, indicating gonadal mosaicism. CONCLUSIONS: The coexistence of CN and SCN phenotypes in this kindred with a shared paternal haplotype strongly suggests both a role for modifying genes in determination of congenital neutropenia disease phenotypes, and the classification of CN and SCN within a spectrum of phenotypes expressing varying degrees of the same disease process.

Probing interactions between CLIP-170, EB1, and microtubules.

Cytoplasmic linker protein 170 (CLIP-170) is a microtubule (MT) plus-end tracking protein (+TIP) that dynamically localizes to the MT plus end and regulates MT dynamics. The mechanisms of these activities remain unclear because the CLIP-170-MT interaction is poorly understood, and even less is known about how CLIP-170 and other +TIPs act together as a network. CLIP-170 binds to the acidic C-terminal tail of alpha-tubulin. However, the observation that CLIP-170 has two CAP-Gly (cytoskeleton-associated protein glycine-rich) motifs and multiple serine-rich regions suggests that a single CLIP-170 molecule has multiple tubulin binding sites, and that these sites might bind to multiple parts of the tubulin dimer. Using a combination of chemical cross-linking and mass spectrometry, we find that CLIP-170 binds to both alpha-tubulin and beta-tubulin, and that binding is not limited to the acidic C-terminal tails. We provide evidence that these additional binding sites include the H12 helices of both alpha-tubulin and beta-tubulin and are significant for CLIP-170 activity. Previous work has shown that CLIP-170 binds to end-binding protein 1 (EB1) via the EB1 C-terminus, which mimics the acidic C-terminal tail of tubulin. We find that CLIP-170 can utilize its multiple tubulin binding sites to bind to EB1 and MT simultaneously. These observations help to explain how CLIP-170 can nucleate MTs and alter MT dynamics, and they contribute to understanding the significance and properties of the +TIP network.