We also performed the following mutations for the amino acid resi

We also performed the following mutations for the amino acid residues surrounding the tryptophans. Because some of the amino acids adjacent to the three tryptophan residues carry electrical charges, we changed the charge in each amino acid residue. We changed two residues, E306 and D308, from acidic to basic amino

acids by replacement with arginine (E306R and D308R). We replaced the residue K310 with glutamic acid in order to change from basic to acidic type (K310E). We also substituted the residue V312 with alanine to maintain hydrophobicity and no electric charge (V312A). We constructed mutant toxins in which we replaced residue N302, the most amino-terminal domain side in the tryptophan-rich region, with alanine (N302A). Wild-type and mutant alpha-toxins were expressed

in E. coli BL21 and purified by affinity chromatography. SDS–PAGE detected every purified mutant toxin at the expected positions selleck chemical and each of their secondary structures was similar to that of wild-type toxin according to far-ultraviolet (190–260 nm) circular dichroism Selleck LEE011 spectral analysis (data not shown). As shown in Table 3, the cytotoxic activities (EC50) of mutant toxins were compared with that of wild-type toxin. We found that the EC50 of W307F/W309F/W311F and W307A/W309A/W311A were >640 ng/mL, indicating that the cytotoxic activity of alpha-toxin decreased remarkably to below the limit of detection. The

mutants of W307A, W309A and W311A also had marked reduction of cytotoxic activity. Although replacements of W307 and W311 with phenylalanine decreased the cytotoxic activities (207 and 113 ng/mL), they did not completely abolish them. Interestingly, replacement of W309 with phenylalanine did not greatly reduce cytotoxic activity. The mutant of W309F retained the same activity as the wild type. In the case of amino acid substitutions surrounding the three tryptophan residues, only D308R caused a decrease in cytotoxic CHIR99021 ability (127 ng/mL). The cytotoxic activities of E306R, K310E, K310R, V312A and N302A did not change in comparison with that of the wild type. To determine whether the tryptophan-rich region plays an important role in the binding of alpha-toxin to cell membranes, we used a toxin overlay assay to examine the binding activities of mutant toxins to detergent-insoluble proteins from Vero cells. After lysis with 1% Triton X-114, we separated Vero cells into detergent-soluble and -insoluble fractions by centrifugation. As shown in Figure 2a, we observed a specific band with a molecular mass of about 34 kDa in the detergent-insoluble fraction using a toxin overlay assay with wild-type alpha-toxin. In previous studies, we reported that alpha-toxin selectively binds to GPI-anchored proteins detected in the detergent-insoluble fractions from various cell lines [12, 25].

Immunoglobulin staining was carried out using Alexa 488-goat anti

Immunoglobulin staining was carried out using Alexa 488-goat anti-mouse κ light chain, FITC or rat anti-mouse IgA (BD-Pharmingen) for 45 min at 37°, then slides XAV-939 mouse were washed in PBS and stained with Dapi for 1 min, Slides or cells were washed in PBS, mounted in Moviol (Merck, Nottingham, UK) and observed on an LSM 510 confocal

microscope (Carl Zeiss, Jena, Germany). Immunohistochemistry was performed on 4-μm paraffin-embedded tissue sections. Samples were pre-treated by microwave incubation in citrate buffer pH6·0 with 0·05% Tween 20. Sections were then incubated for 2 hr at room temperature with the following antibodies: anti-mouse B220 (clone RA3 6B2; BD Biosciences) or anti-CD138 (clone 281-2; BD Biosciences), 1 : 50 in Tris-buffered saline/0·05% Tween. A secondary horseradish peroxidase-conjugated rabbit anti-rat IgG (Dako) was used to reveal primary antibodies for 45 min at room temperature. Acquisitions were carried out on a Zeiss LSM 510 microscope and then analysed with the Image J software (National Institutes of Health, Bethesda, MD) as follows: the complete tissue section surface was measured using the threshold tool; in

the same way, but using a higher threshold, positive staining (B220+ or CD138+ total surface) selleck screening library was evaluated on each section. Finally ratios of B220+ : CD138+ stained areas were calculated. Results are expressed as mean ± SEM (standard error of the mean), and overall differences between variables were evaluated by a two-tailed unpaired Student’s t-test using Prism GraphPad software (Graphpad, San Diego, CA). To block expression of mIgA in B cells, the gene portion encoding the Cα membrane anchoring domain was deleted within the IgH locus (Fig. 1). A neor cassette flanked with loxP sites was inserted as a replacement of the Cα gene membrane exon and was then removed by mating mutants with the Cre transgenic TCL mice (Fig. 1, middle and bottom). Early B-cell compartments in

mutant mice were analysed by flow cytometry. In comparison with wt mice, early B-cell maturation appeared normal in αΔtail+/+ mice. The total number of bone marrow lineage B220+ cells was similar to that in wt controls (26·67 ± 5·085, n = 3 for wt, 21·13 ± 3·839, n = 3 for αΔtail+/+), IgM/IgD expressing cells in αΔtail+/+ bone marrow was similar to that in wt (Fig. 2a) and showed normal absolute values for the CD117+/B220+ pro-B compartment, the CD43+/B220+ pro-B/early pre-B compartment and the B220+/CD25+ pre-B compartment (data not shown). In the periphery, the B220+ cells were similar to wt controls (62·90 ± 0·8591, n = 6 for wt, 67·46 ± 2·152, n = 5 for αΔtail+/+) and the homozygous mutation did not affect the number of surface IgM/IgD expressing cells in the spleen (Fig. 2b).

Tumor microenvironments are disturbed with abnormal growth and re

Tumor microenvironments are disturbed with abnormal growth and remodeling of blood and lymphatic vessels. More effective targeting strategies for delivering anti-angiogenic and cytotoxic agents are being developed through advances in intravital imaging. Blood flow control requires both vasodilation and vasoconstriction

to be coordinated along and among arterioles and feed arteries. Evolving insights into signaling pathways between smooth muscle cells and endothelial cells illuminate how such processes can be affected in vasculopathies. These timely reviews provide a novel reference for advancing research frontiers in microcirculation. “
“The pulmonary circulation is a low-pressure, low-resistance vascular bed with little to no resting tone under normal conditions. IWR-1 mw An increase in the [Ca2+]i in PASMCs is an important determinant of contraction, migration, and proliferation. Both Ca2+ influx through plasma membrane Ca2+ channels and Ca2+ release from the SR contribute to a rise in [Ca2+]i. Additionally important in the pulmonary circulation are several kinase-mediated signaling pathways that act to increase the sensitivity of the contractile apparatus to [Ca2+]i. Similarly, cytoskeletal processes resulting in dynamic remodeling of the actin cytoskeleton can further contribute to contractility in the pulmonary circulation. In addition to endocrine, paracrine, and autocrine factors,

alveolar hypoxia is an important stimulus for pulmonary vasoconstriction. However, prolonged hypoxia is a critical pathological stimulus associated with the development of pulmonary hypertension and cor see more pulmonale. In this review, we will discuss recent advances in our understanding of how Ca2+ homeostasis and sensitization regulate PASMC contractility under both physiological and pathophysiological conditions. “
“Please cite this paper as: Drummond GB, Vowler SL. Variation: use it or misuse it

– replication and its variants. Microcirculation 19: 468–471, 2012. “
“The cerebral blood supply is delivered by a surface network of pial arteries and arterioles from which arise (parenchymal) arterioles that penetrate into the cortex and terminate in a rich capillary bed. The critical regulation of CBF, locally and globally, requires precise vasomotor regulation of the intracerebral microvasculature. This selleck vascular region is anatomically unique as illustrated by the presence of astrocytic processes that envelope almost the entire basolateral surface of PAs. There are, moreover, notable functional differences between pial arteries and PAs. For example, in pial VSMCs, local calcium release events (“calcium sparks”) through ryanodine receptor (RyR) channels in SR membrane activate large conductance, calcium-sensitive potassium channels to modulate vascular diameter. In contrast, VSMCs in PAs express functional RyR and BK channels, but under physiological conditions, these channels do not oppose pressure-induced vasoconstriction.

In addition to CD8+ IELs, the gut also hosts γδTCR T cells, NKT c

In addition to CD8+ IELs, the gut also hosts γδTCR T cells, NKT cells, and classical CD4+ T cells with αβTCR. The exact immune function of all these cells is unknown. The general tendency of these lymphocytes is to generate a tolerogenic immune response to antigens encountered in the gut lumen (20, 21). Other cellular types also participate in mounting an immune response. The most important for promoting oral tolerance are dendritic cells in the lamina propria, which infiltrate the area between

the latero-basal sides of the enterocytes and reach into the intestinal lumen with their projections, taking up antigens which are afterwards processed and presented into the mesenteric lymph nodes (22). Another important cell PARP inhibitor cancer is the so-called M cell, placed as a hood over the luminal region of the PP. These M cells are in contact with PS341 the gut content at their upper pole, allowing them to capture antigens and pass them over to the

immune milieu of the PP, where they are processed by other dendritic cells and then presented to lymphocytes in the local lymph nodes (23). It has been proved that a large proportion of intestinal dendritic cells express an enzyme called retinal dehydrogenase, (responsible for vitamin A metabolism), which produces a shift toward a tolerogenic phenotype in the case of the T helper cells that interact with these dendritic cells (24, 25). All these particularities of the enteric immune system result in generation, at the intestinal level, of Th regulatory cells, also known as iTreg, Tr1, Th3 and Th2 (26). Although intestinal T regulatory cells Ribonucleotide reductase are classical CD4+CD25+FoxP3+ regulatory cells, they appear in

the intestine, and not in the thymus (27). Tr1 (CD4+ IL-10+ FoxP3-) are regulatory cells which exert their function especially through the synthesis of IL-10, while Th3 (CD4+ TGF-β+ FoxP3+) rely on the release of TGF-β for the down regulation of immune responses. These regulatory subpopulations present numerous interconnections in vivo, probably leading to the existence of intermediate cellular types (28). All these characteristics make the gut a predominantly tolerogenic immune environment. The oral administration of any peptide can have three consequences: the secretion of anti-peptide IgA; a systemic immune response with the appearance of serum antibodies and cell-mediated immunity; or a state of anergy, local and/or general tolerance, which prevents an unwanted immune response when re-encountering an innocuous antigen. The first two situations are encountered in the case of pathogens with invasive potential, while the third possibility applies to commensal bacteria and dietary antigens, which do not cause local injuries or systemic immune responses (29).

Cardiac ultrasound and electrocardiography (ECG) should be perfor

Cardiac ultrasound and electrocardiography (ECG) should be performed accordingly, as late-onset cardiotoxicity is described [143]. Thorough monitoring and vigilance is especially relevant for TRAL, as secondary leukaemia is potentially curable if diagnosed early and treated adequately [144], but is associated with potentially fatal complications [145-147] if overlooked. Discussions about SADR incidence, especially TRAL and cardiotoxicity [36, 37, 137, 138, 142, 148-152],

have led to reassessment of the proper risk–benefit profile of MX. TRAL incidences BAY 80-6946 chemical structure vary from 0·07% [149] to 2·82% [138] and are subject to methodological difficulties (e.g. reporting bias especially for meta-analyses [36, 149] and largely lacking prospective data). Interestingly, there seem to be regional differences of TRAL incidence with similar German and French estimates [37, 142], but higher Italian and Spanish rates [137, 138]. Estimates of the incidence

of cardiotoxicity are complicated by different definitions of an adverse cardiac event [reporting of clinical events versus paraclinical abnormalities in ECG, transthoracal echocardiography (TTE) [153] and radionuclide ventriculography [143, 150, 154]]. Subclinical decrease of left ventricular ejection fraction (LVEF) in TTE may be a dose-dependent effect [153]; however, this has not been confirmed by a study with 14% incidence of LVEF decrease in radionuclide ventriculography without dose-dependency [150]. Data on recovery and prognosis of cardiac events are inconsistent [143, 150, 151, 153, 155, 156]. Clinical and paraclinical parameters selleck chemical for the prediction of MX response have been

established [157]. SADR development might be associated with pronounced or lasting leucopenia before TRAL onset [37] and increased brain natriuretic peptide (BNP) in subclinical myocardial injury [158]. In addition to treatment-related factors, genetic factors (genes involved in detoxification: CYP3A4; cellular drug efflux: ABCB1, ABCG2; DNA repair: BRCA2, XRCC5) may influence susceptibility for SADRs [139, 155, 159]. Pharmacogenetic Carnitine palmitoyltransferase II approaches may help early identification of patients at higher risk for side effects or even individualized treatment schemes. The growing spectrum of treatment options for neuroimmunological diseases confronts us with complex risk–benefit considerations and treatment decisions. Whereas established first-line DMDs such as interferon-beta formulations and glatirameracetate are generally safe, newly emerging DMDs with higher efficacy often carry a higher potential of adverse effects with thorough therapy monitoring requirements. Long monitoring intervals, even after cessation of therapy, also pose new challenges for adherence to respective protocols. If not in the clinical trial setting (FTY, alemtuzumab), post-marketing experience (NAT) has revealed relevant or even completely new safety issues not anticipated previously.

The increased acceptance of the elderly with comorbidities, nursi

The increased acceptance of the elderly with comorbidities, nursing home Cetuximab patients with their inherent poor outcomes emphasizes the importance of supporting end-of-life

decisions with palliative care. There should be an associated focus on adequate symptom control, which has been poorly attended to in ESKD as evidenced from some studies. The strong emotional influence, including grief and loss, apparent in the literature for patients, family and health professionals, suggests that there is a real need for education and support in relation to palliative care planning for each of these groups. To do this effectively further rigorous studies are needed to provide a stronger evidence base upon which to advise patients and their families when faced with impending RG7422 dialysis. Some

countries such as the UK, USA, Italy and Canada are well advanced in providing treatment guidelines and resources once dialysis withdrawal is planned but a greater focus on the pre-dialysis phase is required. Multidisciplinary nephrology teams must ensure that patients and their families are accurately informed so they can choose between dialysis and conservative treatment supported by palliative care. The inclusion of palliative care guidelines for Australian nephrology through the CARI guidelines should be considered. The National Health and Medical Research Council is the funder of this study through Grant B0016419. “
“Physical inactivity is a modifiable risk factor for cardiovascular disease. However, the relationship between physical activity and Methocarbamol risk of end-stage kidney disease (ESKD) is not clear. We analyzed

data on a prospective cohort of 59,552 Chinese adults aged 45-74 years enrolled in the Singapore Chinese Health Study. Information on physical activity was collected with a structured questionnaire. Physically active individuals were defined as those who engaged in any moderate activities for 2 hours or more per week, and any strenuous activities 30 minutes or more per week. Incident ESKD was identified via record linkage with the Singapore Registry of Birth and Death and Singapore Renal Registry. Cox proportional hazards regression method was used for analysis for risk of incident ESKD alone or ESKD plus death associated with physical activity. Multivariable models were used to account for the potential confounding effect of sociodemographic, life style factors, and known co-morbidites on the physical activity-ESKD risk association. During a median follow-up of 15.3 years, a total of 642 incident ESKD occurred, and 9808 study participants died. A 24% lower adjusted risk of ESKD [hazard ratio (HR): 0.76; 95% confidence interval (CI): 0.62-0.93] was associated with moderate or strenuous physical activities compared to no regular physical activity. This association appeared to be dose dependent with the lowest risk for subjects at highest intensity of physical activity (p trend <0.003).

6) Interestingly, high levels of IL-22 were also detected

6). Interestingly, high levels of IL-22 were also detected

in the Ivacaftor cell line serum samples of individuals with latent (P = 0·002) and active TB infection (P = 0·003) compared to healthy controls (Fig. 6). IL-1β concentrations in serum of individuals with latent TB infection were increased significantly compared to healthy individuals (P = 0·02). The levels of IL-1β were also higher in individuals with active TB infection but were not statistically significant. Significantly elevated levels of IL-8 were detected in the serum of individuals with latent TB infection only. Mean IL-8 concentrations were significantly higher in latent TB group compared to healthy controls (P < 0·0001). However, the levels of IL-8 were higher but not statistically significant in individuals with active TB infection when compared to healthy individuals (Fig. 6); there was

no difference in the circulating levels of IL-17, IFN-γ (Fig. 6), IL-12p70, IL-2 and TNF-β (data not shown) in serum samples of healthy, latent and active TB subjects. The mean levels of IL-4 in serum of individuals with latent and active TB infection were significantly higher (P = 0·02) than the levels found in healthy subjects (Fig. 6). Levels of IL-5 and IL-10 cytokines were below the detection limit in both antigen-stimulated PBMC culture supernatants as well as in serum samples in all three groups of individuals (data not shown). The present study demonstrates find more differential induction of IFN-γ-, IL-17- and IL-22-expressing CD4+ T cells in Parvulin circulation and following specific stimulation with mycobacterial antigens in TST-negative healthy controls, TST-positive latent and active TB subjects. While the expression of IFN-γ and other cytokines has been analysed in human plasma and PBMC supernatants ex vivo[32,33], the levels of IL-17- and IL-22-expressing CD4+ T cells

and granulocytes in the whole blood of TB patients is not well reported. Herein, we show that the percentage of individuals with active TB expressing IL-17-, IL-22- and IFN-γ-producing CD4+ T cells were decreased significantly compared to the individuals with latent TB infection and healthy controls (Fig. 1). However, such differences were not found in CD8+ T cells (data not shown). The reasons for the decreased IFN-γ-, IL-17- and IL-22-expressing CD4+ T cells in the circulation remain unclear. The differential expression of cytokines in circulation and in affected tissues such as lungs, spleen and lymph nodes have been described in tuberculosis [23,34]. It is possible that antigen-specific IFN-γ-, IL-17- and IL-22-producing CD4+ T cells are recruited to the affected tissues by chemokines released by infected resident macrophages and dendritic cells.

It has become clear that plasma cells are not all alike Plasma c

It has become clear that plasma cells are not all alike. Plasma cells differ in their lifespan, differentiation route, the nature of the produced Ig and their anatomical location [1]. The exact pathways that result in different types of plasma cells are not fully understood, but are suggested to depend on which B cell subset the plasma cells are derived from and which

type of signals are needed to stimulate their differentiation [1, 2]. The B1 cells, marginal zone B cells and follicular B cells can all give rise to plasma cells when activated. The differentiation Trametinib of these cells is a complex process and involves integration of extracellular stimuli to the highly interacting network of transcription factors. The differentiation of B2 cells into antibody-secreting plasma cells can occur via two prominent routes. The cells either differentiate along extrafollicular pathway, creating short-lived plasma cells that produce low-affinity antibodies or proceed to the follicular pathway to generate germinal centres (GCs) that support the maturation of antibody affinity and Ig class switching and long-lived plasma cells (Fig. 1). The type of antigen, the cellular niche and the affinity of BCR towards an antigen determine which differentiation GDC-0980 molecular weight route is chosen with

higher-affinity antigen recognition giving rise to extrafollicular pathway and B cells with lower affinity start to form GCs [3]. Type Thiamine-diphosphate kinase II antigens, which usually contain repeating antigen determinants on a large polysaccharide backbone, can initiate the extrafollicular pathway. The plasma cells from the extrafollicular pathway are sustained in regions such as splenic extrafollicular foci and lymph node medullary chords where CD11chigh dendritic cells provide a proliferation-induced ligand (APRIL) and B cell activating factor (BAFF) [4]. Depending on the subtype, these plasma cells have a half-life ranging from hours to days and usually secrete IgM class antibody and to a lower extent

other Ig classes. The follicular pathway is related to GCs, a specialized structure to support affinity maturation and class switching of Ig. This follicular pathway is known to produce long-lived high-affinity plasma cells that find their survival niches in the bone marrow where they can survive for longer periods [5]. The response to extracellular stimuli and the ability to undergo differentiation are ultimately dictated by transcription factors. The differentiation of B cells into plasma cells involves a substantial change of the gene expression programme, including the repression of B cell transcription factors and other B cell properties [15] as well as induction of plasma cell transcription factors responsible for properties such as active Ig secretion and cessation of cell cycle.

Despite the large geographic distance between Angola and the othe

Despite the large geographic distance between Angola and the other known locations of MVD, phylogenetic analysis using the complete viral genome sequences put Angolan strains within the same clade as the majority of east African isolates [22]. Whereas CFR for MVD are variable (Table 2), the MARV-Angola strain is thought to be more pathogenic than other MARV strains such as the Musoke strain [23-25]. There has been an increase in EVD outbreaks in Africa, probably as result of increased contact between humans and wildlife because of extensive deforestation, hunting and mining [14]. Ebolavirus species have complete genome sequence divergence of 30–45% [7]. The

CFRs of the different ebolavirus species causing these EVD outbreaks have www.selleckchem.com/products/sch772984.html also varied (Table 3). Ebola virus representing the species Zaire ebolavirus can cause sporadic infections in humans, usually resulting in self-limiting outbreaks [26]. The genetic diversity between EBOV strains so far isolated is low [27]. For instance, two separate outbreaks caused by EBOV occurred in Luebo in the DRC in 2007 and 2008: the sequences of the viruses in these two outbreaks were almost identical and related to previously isolated strains, including the one causing the first reported outbreak in Yambuku in the DRC in 1976 [28]. Most recently, there was an outbreak of hemorrhagic fever

caused Tyrosine Kinase Inhibitor Library concentration by EBOV in the West African countries of Guinea, Liberia and Sierra Leone. Full genome sequences of EBOV from three patients showed 97% nucleotide

sequence identity to DRC and Gabon strains of EBOV [29, 30]. TAFV, an ebolavirus belonging to a different species (namely, Taï Forest ebolavirus) Glycogen branching enzyme has been found in the Taï Forest, Côte d’Ivoire [6]; however, the outbreak in West Africa was the first ever reported incidence of EBOV infection in this region [31]. In the 2001–2004 EVD outbreaks in the RC and Gabon, nonhuman primates were also affected by EBOV infections, a large decline occurring in their populations just before and during the outbreaks in humans in the same area [10, 32]. A large serological survey during the 2001–2002 outbreak in Gabon found that dogs might be asymptomatically infected with EBOV, probably as a result of eating infected carcasses or licking body fluids from infected patients, and might potentially transmit EBOV infections [33]. As opposed to EBOV, SUDV, representing the species Sudan ebolavirus, is much more confined geographically, all outbreaks having occurred within a 640 km range [27]. Genetic diversity between the different SUDV strains is very low [27]. In 2011, 7 years after its last appearance, there was a fatal case of SUDV infection in Uganda; the full-length genome sequence of the isolate showed 99.3% identity to the one that caused the Gulu outbreak in 2000 [34].

The differentiating

The differentiating click here step between synthesis of chondroitin suphate and dermatan sulphate GAGs is the epimerization of GlcA to its stereoisomer iduronic acid, whereby the presence of iduronic acid confers synthesis of dermatan sulphate. If alternatively spliced variants of a protein possess GAG initiation sites, they may be referred to as ‘part-time’ proteoglycans [43]. GAG chains are additionally variably sulphated by sulpherotransferase enzymes. CSPGs will be

described in more detail due to their particular relevance to CNS plasticity and repair. CSPGs are a well-studied family of CNS ECM molecules. They are known to play an important role in preventing nerve growth and restricting plasticity following CNS injury and, as such, have been widely targeted in experimental strategies to promote repair in a number of experimental models [44–46]. Members of the CSPG family that are implicated in the response to CNS injury include the lecticans, NG2, phosphacan and the small leucine-rich proteoglycans decorin and biglycan. CS-GAG chains are sulphated at particular residues to form distinct motifs (see Figure 1B). In mammals GalNAc may be mono- or disulphated at C4 and/or C6 to produce chondroitin sulphate-A

selleckchem (CS-A; GlcA-4SGalNAc), chondroitin sulphate-C (CS-C; GlcA-6SGalNAc) and chondroitin sulphate-E (CS-E; GlcA-4S,6SGalNAc) or disulphated at C2 of GlcA and C6 of GalNAc to produce chondroitin sulphate-D (CS-D; GlcA-2S, 6SGalNAc). Chondroitn sulphate-B (CS-B) is dermatan sulphate [47,48]. Sulphation motifs bestow distinct interactive properties upon CSPGs and within PNNs, for example CS-E is specifically thought to provide an attachment site for the guidance cue semaphorin 3A [49,50]. The lecticans next (also known as hyalectan) are the most abundant family of CSPGs within the CNS, comprising aggrecan, versican, neurocan and brevican. Lectican core proteins range in size from 145 kDa to over 300 kDa. They all possess an N-terminal G1 domain and C-terminal G3 domain (see Figure 1C). The G1 domain contains a HA-binding region and immunoglobulin-like

loop, interacting with HA and link protein to form stable ternary complexes in the ECM. The G3 domain comprises EGF repeats (both an EGF module and a calcium-binding EGF module), a C-type lectin domain (CLD) and a complement binding protein-like motif. The CLD has conserved expression across all lecticans and is involved in mediating interactions with other matrix components. This includes ligands with multimeric affinity to CLD such as tenascins, thus thought to enable assembly of cross-linked matrices [51]. Affinity of such interactions may also be regulated by alternative splicing of other G3 domains [52]. Aggrecan additionally includes a G2 domain which is of similar composition to the tandem repeats within G1, but not thought to impart additional interaction with HA [53,54].