Effects of LL-37 on Gingival Fibroblasts: A Role in Periodontal Tissue Remodeling?

Mounting evidence suggests that the host defence peptide, LL-37, plays a role in both inflammation and in wound healing; however, the role of this peptide in the remodeling and maintenance of oral tissues is not yet fully understood. Fibroblasts are the most abundant cell type within the periodontal tissues, and gingival fibroblasts play an important role in maintaining and repairing the gingival tissues which are constantly exposed to external insults. In this study we examined the direct effects of LL-37 treatment on gingival fibroblasts and found that LL-37 significantly increased secretion of both interleukin 8 (IL-8) and IL-6 from these cells. LL-37 tended to decrease matrix metalloproteinase (MMP) activity in gingival fibroblasts, but this decrease did not reach statistical significance. LL-37 significantly increased tissue inhibitor of metalloproteinase-1 (TIMP-1) production by gingival fibroblasts, but had no significant effect on TIMP-2 levels. LL-37 was also shown to significantly increase production of basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and keratinocyte growth factor (KGF) in gingival fibroblasts. Taken together, these results suggest an important role for the host defence peptide, LL-37, in modulating the fibroblast response to remodeling in periodontal tissues

Economic and Social Rights in Northern Ireland: Models of Enforceability

Economic, social and cultural rights (ESR) are those rights defined as such in the International Covenant on Economic, Social and Cultural Rights (1966), the Council of Europe’s Social Rights Charter, the EU’s Charter of Fundamental Rights, and other equivalent legal provisions. In this report, we outline five models for enforcement of economic and social rights (ESR). We use the term ‘model’ to describe these, not in the sense that they are ‘models of best practice’, but simply to indicate that there are various methods already developed which differ from each other in significant ways. There is already extensive, if patchy, implementation of various economic and social rights in Northern Ireland law, even if these protections are not labelled as such. In this context, we need to take into account both common law and statutory provisions regarding rights in the housing, social security, education, employment, human rights, and equality contexts. All of these go some way towards meeting some aspects of internationally-protected ESR, but taken together they still fall short of protecting all internationally-protected ESR to the degree required to satisfy international obligations, as any of the recent reports on the state of ESR in Northern Ireland by the Committee on Economic, Social, and Cultural Rights makes clear. The existing protections do mean, however, that any new initiative is not starting from scratch, which has implications for how best to proceed. The models we discuss below should be regarded as additional to the construction of complementary mechanisms, in civil society particularly, to better enable existing rights that directly or indirectly protect ESR rights, to be implemented more effectively. In particular, it will be important to consider ways in which existing rights could be better mobilised to serve the goal of securing the effective protection of ESR

Biodentine Reduces Tumor Necrosis Factor Alpha-induced TRPA1 Expression in Odontoblastlike Cells

International audienceIntroduction: The transient receptor potential (TRP) ion channels have emerged as important cellular sensors in both neuronal and non-neuronal cells, with TRPA1 playing a central role in nociception and neurogenic inflammation. The functionality of TRP channels has been shown to be modulated by inflammatory cytokines. The aim of this study was to investigate the effect of inflammation on odontoblast TRPA1 expression and to determine the effect of Biodentine (Septodent, Paris, France) on inflammatory-induced TRPA1 expression. Methods: Immunohistochemistry was used to study TRPA1 expression in pulp tissue from healthy and carious human teeth. Pulp cells were differentiated to odontoblastlike cells in the presence of 2 mmol/L beta-glycerophosphate, and these cells were used in quantitative polymerase chain reaction, Western blotting, calcium imaging, and patch clamp studies. Results: Immunofluorescent staining revealed. TRPA1 expression in odontoblast cell bodies and odontoblast processes, which was more intense in carious versus healthy teeth. TRPA1 gene expression was induced in cultured odontoblastlike cells by tumor necrosis factor alpha, and this expression was significantly reduced in the presence of Biodentine. The functionality of the TRPA1 channel was shown by calcium microfluorimetry and patch clamp recording, and our results showed a significant reduction in tumor necrosis factor alpha induced TRPA1 responses after Biodentine treatment. Conclusions: In conclusion, this study showed TRPA1 to be modulated by caries-induced inflammation and that Biodentine reduced TRPA1 expression and functional responses

Identification and pharmaceutical evaluation of novel frog skin-derived serine proteinase inhibitor peptide–PE-BBI (Pelophylax esculentus Bowman-Birk inhibitor) for the potential treatment of cancer

Abstract Amphibian venom-derived peptides have high potential in the field of anticancer drug discovery. We have isolated a novel Bowman-Birk proteinase inhibitor (BBI)-type peptide from the skin secretion of Pelophylax esculentus (PE) named PE-BBI, and evaluated its bio-functions and anti-cancer activity in vitro. PE-BBI is a heptadecapeptide with C-terminal amidation. The mRNA sequence and primary structure of PE-BBI were identified using RT-PCR and LC/MS, respectively. A trypsin inhibitory assay was used to characterize the serine proteinase inhibitory activity of synthetic PE-BBI. PE-BBI’s myotropic activity was analyzed using isolated rat bladder and rat-tail artery smooth muscle tissues, and the anti-cancer ability of PE-BBI using human colorectal cancer cells. PE-BBI’s mechanism of action was investigated using Discovery studio software. PE-BBI showed trypsin inhibitory activity (Ki = 310 ± 72 nM), strong myotropic activity, and cytotoxicity that were specific to cancer cells, and no side effect to normal epithelial cells. The docking stimulation showed that PE-BBI had high affinity to several members of human kallikrein related peptidase (KLK) family. This finding helps to enrich our understanding of BBI peptides’ mode of action. Moreover, the data presented here validates frog secretions as sources of potential novel proteinase inhibitors for cancer treatment

Vasoactivity of Rucaparib, a PARP-1 Inhibitor, is a Complex Process that Involves Myosin Light Chain Kinase, P2 Receptors, and PARP Itself

Therapeutic inhibition of poly(ADP-ribose) polymerase (PARP), as monotherapy or to supplement the potencies of other agents, is a promising strategy in cancer treatment. We previously reported that the first PARP inhibitor to enter clinical trial, rucaparib (AG014699), induced vasodilation in vivo in xenografts, potentiating response to temozolomide. We now report that rucaparib inhibits the activity of the muscle contraction mediator myosin light chain kinase (MLCK) 10-fold more potently than its commercially available inhibitor ML-9. Moreover, rucaparib produces additive relaxation above the maximal degree achievable with ML-9, suggesting that MLCK inhibition is not solely responsible for dilation. Inhibition of nitric oxide synthesis using L-NMMA also failed to impact rucaparib's activity. Rucaparib contains the nicotinamide pharmacophore, suggesting it may inhibit other NAD+-dependent processes. NAD+ exerts P2 purinergic receptor-dependent inhibition of smooth muscle contraction. Indiscriminate blockade of the P2 purinergic receptors with suramin abrogated rucaparib-induced vasodilation in rat arterial tissue without affecting ML-9-evoked dilation, although the specific receptor subtypes responsible have not been unequivocally identified. Furthermore, dorsal window chamber and real time tumor vessel perfusion analyses in PARP-1-/- mice indicate a potential role for PARP in dilation of tumor-recruited vessels. Finally, rucaparib provoked relaxation in 70% of patient-derived tumor-associated vessels. These data provide tantalising evidence of the complexity of the mechanism underlying rucaparib-mediated vasodilation

Microneedle-mediated vaccine delivery:harnessing cutaneous immunobiology to improve efficacy

INTRODUCTION: We describe the use of microneedle arrays for delivery to targets within the skin itself. Breaching the skin’s stratum corneum barrier raises the possibility of administration of vaccines, gene vectors, antibodies and even nanoparticles, all of which have at least their initial effect on populations of skin cells. AREAS COVERED: Intradermal vaccine delivery, in particular, holds enormous potential for improved therapeutic outcomes for patients, particularly those in the developing world. Various vaccine-delivery strategies have been employed and here we discuss each one in turn. We also describe the importance of cutaneous immunobiology on the effect produced by microneedle-mediated intradermal vaccination. EXPERT OPINION: Microneedle-mediated vaccine holds enormous potential for patient benefit. In order for microneedle vaccine strategies to fulfil their potential, however, the proportion of an immune response that is due to local action of delivered vaccines on skin antigen presenting cells and what is due to a systemic effect from vaccine reaching the systemic circulation must be determined. Moreover, industry will need to invest significantly in new equipment and instrumentation in order to mass produce microneedle vaccines consistently. Finally, microneedles will need to demonstrate consistent dose delivery across patient groups and match this to reliable immune responses before they will replace tried- and-tested needle-and-syringe based-approaches

Vasoactivity of AG014699, a clinically active small molecule inhibitor of poly(ADP-ribose) polymerase: a contributory factor to chemopotentiation in vivo?

PURPOSE: Poly(ADP-ribose) polymerase plays an important role in DNA repair and PARP inhibitors can enhance the activity of DNA damaging agents in vitro and in vivo. AG014699 is a potent PARP inhibitor in Phase II clinical development. However the range of therapeutics with which AG014699 could interact via a DNA-repair based mechanism is limited. We aimed to investigate a novel, vascular-based activity of AG014699, underlying in vivo chemosensitisation, that could widen its clinical application. EXPERIMENTAL DESIGN: Temozolomide response was analysed in vitro and in vivo. Vessel dynamics were monitored using “mismatch” following the administration of perfusion markers and real-time analysis of fluorescently-labeled albumin uptake in to tumours established in dorsal window chambers. Further mechanistic investigations employed ex vivo assays of vascular smooth muscle relaxation, gut motility and myosin light chain kinase inhibition. RESULTS: AG014699 failed to sensitise SW620 cells to temozolomide in vitro but induced pronounced enhancement in vivo. AG014699 (1mg/kg) improved tumour perfusion comparably with the control agents nicotinamide (1g/kg) and AG14361 (fore-runner to AG014699; 10mg/kg). AG014699 and AG14361 relaxed pre-constricted vascular smooth muscle more potently than the standard agent, hydralazine, with no impact on gut motility. AG014699 inhibited myosin light chain kinase at concentrations that relaxed isolated arteries, whereas AG14361 had no effect. CONCLUSION: Increased vessel perfusion elicited by AG014699 could increase tumour drug accumulation and therapeutic response. Vasoactive concentrations of AG014699 do not cause detrimental side-effects to gut motility and may increase the range of therapeutics with which AG014699 could be combined with for clinical benefit

Effects of rucaparib on tumor vessel perfusion may be dependent on PARP.

<p>Panels A and B; B16 tumours were established in PARP WT or KO female mice. The extent of vessel ‘mismatch’ following the administration of the perfusion markers Hoechst 33342 and carbocyanin was reduced by rucaparib (1 mg/kg) in tumors established in WT but not PARP-1^-/- mice. Panel C; fold change in intratumoral fluorescence above that seen following initial plateau (20 min) in dorsal window chambers implanted with B16 tumors in WT and PARP-1^-/- mice and treated with 1 mg/kg rucaparib. Panel D; representative real time analysis of the accumulation of BSA-647 (administered via iv injection at time 0) in B16 tumors established in dorsal window chambers in PARP WT (closed symbols) and PARP-1^-/- (open symbols) mice. Arrow indicates the administration of rucaparib (10 mg/kg). NS—p >0.05, *p<0.05, **p<0.01 as compared with relevant control. N = 3 mice per condition.</p