The neutralising and stimulatory effects of antimicrobial peptide LL-37 in human gingival fibroblasts.

OBJECTIVES: To investigate the effects of LL-37, a broad spectrum antimicrobial peptide expressed in periodontal tissues, on human gingival fibroblast responsiveness to microbial challenge and to explore the direct effects of LL-37 on human gingival fibroblasts. DESIGN: The effect of LL-37 on bacterial lipopolysaccharide-induced expression of Interleukin (IL-6) and chemokine C-X-C motif ligand (CXCL) 8 was determined by enzyme linked immunosorbent assay (ELISA). LL-37's influence on bacterial lipopolysaccharide-induced IκBα degradation was investigated by western blot. DNA microarray analysis initially determined the direct effects of LL-37 on gene expression, these findings were subsequently confirmed by quantitative polymerase chain reaction and ELISA analysis of selected genes. RESULTS: Bacterial lipopolysaccharide-induced IL-6 and CXCL8 production by human gingival fibroblasts was significantly reduced in the presence of LL-37 at concentrations in the range of 1-10 µg/ml. LL-37 led to a reduction in lipopolysaccharide-induced IκBα degradation by Escherichia coli lipopolysaccharide and Porphyromonas gingivalis lipopolysaccharide (10 µg/ml). LL-37 (50 µg/ml) significantly altered the gene expression of 367 genes in human gingival fibroblasts by at least 2-fold. CXCL1, CXCL2, CXCL3, Interleukin-24 (IL-24), CXCL8, Chemokine (C-C motif) Ligand 2, and Suppressor of Cytokine Signalling 3 mRNA were significantly upregulated by LL-37. LL-37 also significantly stimulated expression of CXCL8, hepatocyte growth factor and CXCL1 at the protein level. CONCLUSION: LL-37 plays an important regulatory role in the immunomodulatory activity of gingival fibroblasts by inhibiting lipopolysaccharide -induced expression of inflammatory cytokines and directly stimulating the expression of an array of bioactive molecules involved in inflammation and repair.

Odontoblast cell death induces NLRP3 inflammasome-dependent sterile inflammation and regulates dental pulp cell migration, proliferation and differentiation.

AIM: To investigate the ability of dead odontoblasts to initiate NLRP3 inflammasome-dependent sterile inflammation and to explore the effect on dental pulp cell (DPCs) migration, proliferation and odontogenic differentiation. METHODS: Odontoblast-like cells were subjected to freezing-thawing cycles to produce odontoblast necrotic cell lysate (ONCL). DPCs were treated with ONCL to assess proliferation and migration. THP-1 differentiated macrophages stimulated with ONCL and live cell imaging and western blotting were used to assess NLRP3 inflammasome activation. Cytokines were measured with multiplex arrays and ELISA. qPCR, alkaline phosphatase and Alizarin red assays were used to assess odontogenic differentiation of DPCs. Data were analysed using the t-test or anova followed by a Bonferroni post hoc test with the level of significance set at P ≤ 0.05. RESULTS: ONCL induced migration and proliferation of DPCs. Treatment of THP-1 macrophages with ONCL resulted in the release of the inflammatory cytokines IL-1ß, IL-6, IL-8, TNFα, IFN-γ, CCL2 and angiogenic growth factors, angiogenin and angiopoietin. This inflammatory response was associated with activation of NFκB, p38MAPK and NLRP3 inflammasome. To confirm that ONCL induced inflammatory response is NLRP3 inflammasome-dependent, treatment with a caspase-1 inhibitor and a specific NLRP3 inhibitor significantly reduced IL-1ß release in THP-1 macrophages (P = 0.01 and 0.001). Inflammasome activation product, IL-1ß, induced odontogenic differentiation of DPCS as evident by the increase in odontogenic genes expression DMP-1, RUNX-2, DSPP and SPP, alkaline phosphatase activity and mineralization. CONCLUSION: Dead odontoblasts induced NLRP3 inflammasome-dependent sterile inflammation and activated the migration, proliferation and differentiation of DPCs.

A connectivity mapping approach predicted acetylsalicylic acid (aspirin) to induce osteo/odontogenic differentiation of dental pulp cells.

AIM: To use connectivity mapping, a bioinformatics approach, to identify compounds that could induce odontogenic differentiation of dental pulp cells (DPCs) and to experimentally validate this effect. A subsidiary aim was to investigate the anti-inflammatory effect of any identified compound. METHODOLOGY: The Gene Expression Omnibus (GEO) database was searched for microarray data sets assessing odontogenic differentiation of human DPCs. An odontogenic gene expression signature was generated by differential expression analysis. The statistical significant connectivity map (ssCMap) method was used to identify compounds with a highly correlating gene expression pattern. DPCs were treated with the compound identified, and osteo/odontogenic differentiation was assessed by Alizarin red staining, alkaline phosphatase activity and expression of osteo/odontogenic genes ALPL, RUNX2, COL1A1, DSPP, DMP1 and SPP1 by RT-PCR. The anti-inflammatory effect of the compound was assessed using an ex vivo pulpitis model, and cytokine levels were measured with multiplex assay. Means were compared using the t-test or ANOVA followed by a Bonferroni post hoc test with the level of significance set at P ≤ 0.05. RESULTS: The GEO database search identified a specific gene expression signature for osteo/odontogenic differentiation. Analysis using ssCMap found that acetylsalicylic acid [(ASA)/aspirin] was the drug with the strongest correlation with that gene signature. The treatment of DPCs with 0.05 mmol L-1 ASA showed increased alkaline phosphatase activity (P < 0.001), mineralization (P < 0.05), and increased the expression of the osteo/odontogenic genes, DMP1 and DSPP (P < 0.05). Low concentration (0.05 mmol L-1 ) ASA reduced inflammatory cytokines IL-6 (P < 0.001), CCL21 (P < 0.05) and MMP-9 (P < 0.05) in an ex vivo pulpitis model. CONCLUSIONS: Connectivity mapping, a web-based informatics method, was successfully used to identify aspirin as a candidate drug that could modulate the differentiation of DPCs. Aspirin was shown to induce odontogenic differentiation in DPCs in vitro and this, together with its anti-inflammatory effects, makes it a potential candidate for vital pulp therapies.

Abdominal Drains Retrieved Laparoscopically 15 Years Post Laparotomy.

A retained intra-abdominal foreign body is a common occurrence that is frequently underreported due to its medicolegal implications. Sponges, gauzes, surgical instruments, abdominal drains, etc. have been reported in the literature. The most common presentation for a retained intra-abdominal foreign body is postoperative abdominal infections or bleeding, frequently seen in the immediate postoperative period. Most of these foreign bodies are removed by exploratory laparotomy owing to recent abdominal surgeries or presentation as complicated abdominal masses. Here, we report a case with retained intra-abdominal drains for 15 years with minimal symptoms presenting as an intermittent abdominal pain; the drains were removed using laparoscopic intervention.

Hernioscopy in Incarcerated Inguinal Hernia Spontaneously Reduced after General Anesthesia Induction.

Hernioscopy is essentially hernia sac laparoscopy. Hernia repair has evolved over the years with better outcomes; however, strangulated inguinal hernias are acute surgical emergencies which require emergent operative intervention. During anesthesia induction and/or after incision, hernia self-reduction is possible, with or without compromised bowel, back into the abdominal cavity. It is pivotal to examine the bowel to decide on further operative course. A simple alternative to unnecessary laparotomy or standard laparoscopy is hernioscopy, which is quite uncommon. We present a case of an acute symptomatic strangulated left-sided inguinal hernia which got self-reduced during anesthesia induction and was successfully repaired after hernioscopy was used to evaluate the incarcerated hernia content. We provide a brief review of literature about hernioscopy and an algorithm to guide surgeons in emergent cases.

Matrix metalloproteinase-8 activity in gingival crevicular fluid: development of a novel assay.

BACKGROUND AND OBJECTIVE: The matrix metalloproteinases (MMPs) play a role in regulating turnover and metabolism of connective tissues in health but they have also been implicated in a wide variety of pathological conditions, including periodontal disease. MMP-8 has been extensively studied in periodontal health and disease using ELISA, although this technique is limited by its inability to determine enzyme activity. The aim was to develop an assay specifically to measure MMP-8 activity and to demonstrate its use in the analysis of gingival crevicular fluid samples. MATERIAL AND METHODS: A specific antibody was used to coat black 96-well microtitre plates to capture MMP-8 selectively. The activity of bound MMP-8 was measured using a fluorogenic substrate. Gingival crevicular fluid samples, from healthy and periodontally diseased sites, were collected using PerioPaper strips and tested for MMP-8 activity. RESULTS: Significantly higher MMP-8 activity was demonstrated in gingival crevicular fluid from periodontally diseased sites compared with healthy sites that exhibited basal or no MMP-8 activity. No cross-reactivity with other MMPs was noted. CONCLUSION: We show, for the first time, that MMP-8 activity can be specifically detected and quantified in gingival crevicular fluid samples. Measurement of MMP-8 activity could prove to be useful in monitoring periodontal disease progression.

p53, Cyclin D1, p21 (WAF1) and Ki-67 (MIB1) Expression at Invasive Tumour Fronts of Oral Squamous Cell Carcinomas and Development of Local Recurrence.

BACKGROUND: Expression of p53, cyclin D1, p21 (WAF1) and Ki-67 (MIB1) was evaluated in oral squamous cell carcinoma (OSCC) to test whether levels of these markers at invasive tumour fronts (ITFs) could predict the development of local recurrence. MATERIALS AND METHODS: Archived paraffin-embedded specimens from 51 patients with T1/T2 tumours were stained immunohistochemically and analysed quantitatively. Local recurrence-free survival was tested with Kaplan-Meier survival plots (log-rank test) using median values to define low and high expression groups and with a Cox's proportional hazards model in which the expression scores were entered as continuous variables. RESULTS: The assessment of expression of all markers was highly reliable, univariate analysis showing that patients with clear surgical margins, with low cyclin D1 and high p21 expression at the ITF had the best local recurrence-free survival. Multivariate analysis showed that these three parameters were independent prognostic factors but that neither p53 nor MIB1 expression were of prognostic value. CONCLUSIONS: Assessment of p53, cyclin D1, p21 (WAF1), and Ki-67 (MIB1) at the ITF could help to predict local recurrence in early stage oral squamous cell carcinoma cases.

Dietary patterns, nutrition knowledge and lifestyle: associations with blood pressure in a sample of Australian adults (the Food BP study).

This study examined the association between dietary patterns, nutrition knowledge and lifestyle with blood pressure (BP) in a sample of Australian adults. Adults with normal and high BP were included in a cross-sectional study. Dietary intake data was collected using a Food Frequency Questionnaire. Nutrition knowledge and lifestyle surveys were included in the questionnaire. Dietary patterns were extracted using factor analysis followed by cluster analysis. Associations were analysed using logistic regression. Four hundred and seven participants were included. Three dietary patterns were identified: Western; Snack and alcohol; and Balanced. Participants with high BP had a higher intake of Western and a lower intake of Balanced dietary pattern. A significant and higher frequency of discretionary foods and oils consumption, as well as lower nutrition knowledge score and activity frequency, were observed in the high BP group. Regression analysis indicated that the intake of Western and Snack and alcohol dietary patterns increases the likelihood of having high BP by 2.40 (95% confidence interval (CI): 1.28-4.49) and 2.76 (95% CI: 1.52-5.00), respectively, when nutrition knowledge and lifestyle were controlled for as moderator variables. The likelihood of high BP was not associated with nutrition knowledge, but increased with physical inactivity. This study indicates that poor dietary patterns and inactivity are associated with increases in the likelihood of high BP, and the association is not influenced by nutrition knowledge. These findings indicate the importance of developing public health strategies with an emphasis on improving the dietary patterns of individuals to prevent and control high BP in Australian adults.

Interferon-γ stimulates CD14, TLR2 and TLR4 mRNA expression in gingival fibroblasts increasing responsiveness to bacterial challenge.

OBJECTIVE: To investigate the potential effects of IFN-03A5 on the responsiveness of human gingival fibroblasts to bacterial challenge. DESIGN: mRNA and protein expression of CD14, TLR2 and TLR4 in human gingival fibroblasts was detected by quantitative polymerase chain reaction (Q-PCR) and flow cytometry. The effect of preincubation with IFN-03A5 on subsequent bacterial LPS-induced expression of IL-6 and IL-8 by gingival fibroblasts was determined by ELISA. Bacterial LPS-induced IκBα degradation in human gingival fibroblasts was investigated by western blot. RESULTS: Human gingival fibroblasts express CD14, TLR2 and TLR4 mRNAs. IFN-03A5, but not IL-103B2, induced mRNA expression of all three receptors and the expression of membrane bound CD14 protein. Pre-incubation of fibroblasts with IFN-03A5 and subsequent stimulation with Escherichia coli LPS or Porphyromonas gingivalis LPS led to increased production of IL-6 and IL-8. LPS-induced pro-inflammatory cytokine production was abrogated by a blocking antibody to CD14. Both E. coli LPS and P. gingivalis LPS induced IκBα degradation in human gingival fibroblasts. CONCLUSION: Our data indicate that IFN-03A5 primes human gingival fibroblasts, through the upregulation of CD14 expression, which results in increased responsiveness to bacterial LPS challenge, as determined by pro-inflammatory cytokine production.

Long-term follow up of paediatric liver transplant recipients: outcomes following transfer to adult healthcare in New Zealand.

Poor outcomes are reported in young people with chronic health conditions. We performed a retrospective notes review of New Zealand paediatric liver transplant recipients transferred to adult services. Two patients were lost to follow up. Out of 20, 12 were non-adherent, and out of 12, 7 developed rejection. Other risk behaviours were common in the non-adherent group. We conclude that dedicated services for these young people may be needed to optimise outcomes.

The wish to be hated: Richard III's villainy in pursuit of the bad-enough object.

Richard III's legendary villainy, in addition to its obvious role in furthering his ambition for power, arguably had deeper psychological roots: motives embedded in the developmental unfolding of his complex relationship with a severely traumatized and rejecting mother. The theoretical formulations of Ronald Fairbairn are used to show that Richard's villainy preserved and perpetuated a specific object-tie to his mother, that of being her demonized and despised "child of hate," and that many of his victims, especially women, served as maternal surrogates in his lifelong quest to replicate his maternal "bad-enough object." Similar patterns of severe relational disturbance pervade many of the mother-son dyads portrayed in Shakespeare's other plays, a repetitive theme that may well be linked to the playwright's own mother's history of severe loss and depression during his infancy and its impact on her ability to provide her son with bountiful emotional nurturance. A collateral issue is also addressed-the narcissistically embedded bonding difficulties between parents and their disabled children, whose visible impairment is said to give rise to greater adjustment problems than does the medical severity of the condition itself.

Development and application of a multi-axis dynamometer for measuring grip force.

OBJECTIVE: This paper describes the development and application of a novel multi-axis hand dynamometer for quantifying 2D grip force magnitude and direction in the flexion-extension plane of the fingers. METHODS: A three-beam reconfigurable form dynamometer, containing two active beams for measuring orthogonal forces and moments regardless of point of force application, was designed, fabricated and tested. Maximum grip exertions were evaluated for 16 subjects gripping cylindrical handles varying in diameter. RESULTS: Mean grip force magnitudes were 231 N (SD = 67.7 N), 236 N (72.9 N), 208 N (72.5 N) and 158 N (45.7 N) for 3.81 cm, 5.08 cm, 6.35 cm and 7.62 cm diameter handles, respectively. Grip force direction rotated clockwise and the centre of pressure moved upward along the handle as handle diameter increased. CONCLUSIONS: Given that the multi-axis dynamometer simultaneously measures planar grip force magnitude and direction, and centre of pressure along the handle, this novel sensor design provides more grip force characteristics than current sensor designs that would improve evaluation of grip characteristics and model-driven calculations of musculoskeletal forces from dynamometer data.

Illustrated Glossary of Radiology Anatomy, Examinations, and Procedures

Glossário ilustrado e multimídia de radiologia: anatomia, exames e outros procedimentos radiológicos. A descrição dos termos oferecem acesso à imagens descritivas e estudos de casos da base de dados MedPix.

The effect of topical phenytoin on healing in diabetic foot ulcers: a randomized controlled trial.

AIM: The aim of the study was to evaluate the effect of topical phenytoin on healing in diabetic foot ulcers. A randomized, controlled, double-blind, clinical trial was conducted. METHODS: A phenytoin dressing and a control dressing were manufactured. Individuals who were ≥ 18 years of age with peripheral neuropathy, an ankle brachial pressure index > 0.5 and a diabetic foot ulcer ≥ 4 weeks' duration were independently randomized to the phenytoin group (31 participants) or the control group (34 participants). Participants with renal disease, ankle brachial pressure index < 0.5, necrosis or osteomyelitis were excluded. Subjects received standard wound care and dressing application. Primary endpoint analysis (diabetic foot ulcer closed or not at 16 weeks) was calculated by survival analysis. RESULTS: Participants (n = 65, 52 with Type 2 diabetes) were treated for a maximum of 16 weeks. Sixty per cent of the diabetic foot ulcers closed overall (18 in the phenytoin group, 20 in the control group) with no statistically significant differences in complete healing or in diabetic foot ulcer area over time between the two groups. At 24-weeks follow-up, one diabetic foot ulcer had recurred. CONCLUSIONS: There were no differences in diabetic foot ulcer closure rates or in diabetic foot ulcer area over time between the two groups. This study does not support the use of phenytoin in the treatment of diabetic foot ulcers.