Melanoma secretion of transforming growth factor-ß2 leads to loss of epidermal AMBRA1 threatening epidermal integrity and facilitating tumour ulceration.

BACKGROUND: For patients with early American Joint Committee on Cancer (AJCC)-stage melanoma the combined loss of the autophagy regulatory protein AMBRA1 and the terminal differentiation marker loricrin in the peritumoral epidermis is associated with a significantly increased risk of metastasis. OBJECTIVES: The aim of the present study was to evaluate the potential contribution of melanoma paracrine transforming growth factor (TGF)-ß signalling to the loss of AMBRA1 in the epidermis overlying the primary tumour and disruption of epidermal integrity. METHODS: Immunohistochemistry was used to analyse AMBRA1 and TGF-ß2 in a cohort of 109 AJCC all-stage melanomas, and TGF-ß2 and claudin-1 in a cohort of 30 or 42 AJCC stage I melanomas, respectively, with known AMBRA1 and loricrin (AMLo) expression. Evidence of pre-ulceration was analysed in a cohort of 42 melanomas, with TGF-ß2 signalling evaluated in primary keratinocytes. RESULTS: Increased tumoral TGF-ß2 was significantly associated with loss of peritumoral AMBRA1 (P < 0·05), ulceration (P < 0·001), AMLo high-risk status (P < 0·05) and metastasis (P < 0·01). TGF-ß2 treatment of keratinocytes resulted in downregulation of AMBRA1, loricrin and claudin-1, while knockdown of AMBRA1 was associated with decreased expression of claudin-1 and increased proliferation of keratinocytes (P < 0·05). Importantly, we show loss of AMBRA1 in the peritumoral epidermis was associated with decreased claudin-1 expression (P < 0·05), parakeratosis (P < 0·01) and cleft formation in the dermoepidermal junction (P < 0·05). CONCLUSIONS: Collectively, these data suggest a paracrine mechanism whereby TGF-ß2 causes loss of AMBRA1 overlying high-risk AJCC early-stage melanomas and reduced epidermal integrity, thereby facilitating erosion of the epidermis and tumour ulceration.

Effect of curing mode on the conversion and IIT-derived mechanical properties of core build-up resin composites.

The aim of the study was to evaluate the degree of conversion and the mechanical properties of five composite core build-up materials polymerized in dual-curing and self-curing modes. The materials tested were: Clearfil DC Core Plus (CF), Gradia Core (GC), Luxacore-Z Dual Smartmix (LX), Multicore Flow (MC) and Paracore (PC). Disk-shaped specimens were prepared from each material; half the specimens were light-cured, whereas the rest were only self-cured. After a 3-week storage period (dark/dry/37 °C) the Martens Hardness, Indentation Modulus, and Elastic Index were determined by instrumented indentation testing (IIT), while the degree of conversion was assessed by ATR-FTIR spectroscopy. Statistical analysis was performed by 2-way ANOVA and post-hoc testing (α = 0.05). The dual-curing mode resulted in statistically higher Martens Hardness and Elastic Index than the self-curing mode in most materials but showed insignificant differences in Indentation Modulus. MC and PC demonstrated significantly higher degree of conversion in both curing modes. Overall, the self-curing mode was inferior to the dual-curing in conversion and mechanical properties for most products, despite their differences in monomer composition and filler loading.

Epidermal autophagy and beclin 1 regulator 1 and loricrin: a paradigm shift in the prognostication and stratification of the American Joint Committee on Cancer stage I melanomas.

BACKGROUND: The updated American Joint Committee on Cancer (AJCC) staging criteria for melanoma remain unable to identify high-risk stage I tumour subsets. OBJECTIVES: To determine the utility of epidermal autophagy and beclin 1 regulator 1 (AMBRA1)/loricrin (AMLo) expression as a prognostic biomarker for AJCC stage I cutaneous melanoma. METHODS: Peritumoral AMBRA1 expression was evaluated in a retrospective discovery cohort of 76 AJCC stage I melanomas. AMLo expression was correlated with clinical outcomes up to 12 years in two independent powered, retrospective validation and qualification cohorts comprising 379 AJCC stage I melanomas. RESULTS: Decreased AMBRA1 expression in the epidermis overlying primary melanomas in a discovery cohort of 76 AJCC stage I tumours was associated with a 7-year disease-free survival (DFS) rate of 81·5% vs. 100% survival with maintained AMBRA1 (P < 0·081). Following an immunohistochemistry protocol for semi-quantitative analysis of AMLo, analysis was undertaken in validation (n = 218) and qualification cohorts (n = 161) of AJCC stage I melanomas. Combined cohort analysis revealed a DFS rate of 98·3% in the AMLo low-risk group (n = 239) vs. 85·4% in the AMLo high-risk cohort (n = 140; P < 0·001). Subcohort multivariate analysis revealed that an AMLo hazard ratio (HR) of 4·04 [95% confidence interval (CI) 1·69-9·66; P = 0·002] is a stronger predictor of DFS than Breslow depth (HR 2·97, 95% CI 0·93-9·56; P = 0·068) in stage IB patients. CONCLUSIONS: Loss of AMLo expression in the epidermis overlying primary AJCC stage I melanomas identifies high-risk tumour subsets independently of Breslow depth. What's already known about this topic? There is an unmet clinical need for biomarkers of early-stage melanoma. Autophagy and beclin 1 regulator 1 (AMBRA1) is a proautophagy regulatory protein with known roles in cell proliferation and differentiation, and is a known tumour suppressor. Loricrin is a marker of epidermal terminal differentiation. What does this study add? AMBRA1 has a functional role in keratinocyte/epidermal proliferation and differentiation. The combined decrease/loss of peritumoral AMBRA1 and loricrin is associated with a significantly increased risk of metastatic spread in American Joint Committee on Cancer (AJCC) stage I tumours vs. melanomas, in which peritumoral AMBRA1 and loricrin are maintained, independently of Breslow depth. What is the translational message? The integration of peritumoral epidermal AMBRA1/loricrin biomarker expression into melanoma care guidelines will facilitate more accurate, personalized risk stratification for patients with AJCC stage I melanomas, thereby facilitating stratification for appropriate follow-up and informing postdiagnostic investigations, including sentinel lymph node biopsy, ultimately resulting in improved disease outcomes and rationalization of healthcare costs.

Biocompatibility of resin composites subcutaneously implanted in rats with experimentally induced arthritis.

OBJECTIVE: To evaluate the biocompatibility of resin composite specimens with different curing efficiency, subcutaneously implanted in rats with experimentally induced arthritis. METHODS: The amount of remaining CC bonds (%RDB) of hybrid resin composite specimens photopolymerized for 10s and 40s exposure time (n=3) was measured by micro-attenuated total reflectance Fourier transform infrared spectroscopy. Male Wistar rats (n=36) were classified in two groups (n=18) of healthy animals and of animals with experimentally induced arthritis. Resin composite specimens irradiated for 10s and 40s and calcium hydroxide control specimens were implanted subcutaneously in each animals' dorsum. Following 2-, 4- and 9-week periods the animals were sacrificed. The development of arthritis was defined by biochemical analysis and the changes in the relative weight of animals' organs (spleen, thymus, adrenals). Tissue reactions were examined histologically. RESULTS: %RDB per site and exposure time showed statistically significant differences. Lowest %RDB values were recorded on 40s exposed specimens. Biochemical indices and relative organ weights demonstrated statistically significant differences between healthy animals and animals with arthritis. The health status of the animals and the materials used did not influence tissue response. First and second periods of sacrifice showed reduced propensity of connective tissue development in comparison to the third period. The same applied for the second period regarding the presence of giant cells. SIGNIFICANCE: The materials tested and the animals' health status did not result in altered tissue response compared to control group. The period of sacrifice was associated with different tissue responses.

Impact of experimental trauma and niflumic acid administration on antimicrobials' concentration in serum and mandible of rats.

Administration of antibiotics and analgesics in surgery or trauma is of great importance for an effective treatment. Trauma, as stress stimulus, causes alterations in various functions of the organism as well as in drug pharmacokinetics. The aim of this study was to determine the effect of trauma upon the serum and bone levels of the antimicrobial ampicillin and cefapirin, with and without co-administration of a non-steroidal anti-inflammatory analgesic (NSAIDs). Fifty-six male Wistar rats were divided into two groups A (control) and B (experimental). Each group consisted of 4 subgroups (n=7) receiving ampicillin, ampicillin with niflunic acid, cefapirin, and cefapirin with niflunic acid. In group B traumatic injury was performed by incision (7 mm length) in the right cheek. The levels of the antibiotics were estimated by the inhibition zone of B. subtilis. An increase in antibiotic levels was observed in group B, being statistically significant only for cefapirin level in the mandible. Upon niflumic acid co-administration a statistically significant rise in serum ampicillin and mandible cefapirin levels was observed in both control and experimental groups (student t-test). It can be concluded that the combination of antibiotics and non-steroid antiinflammatory drugs (NSAIDs) may enhance the antibacterial drug concentration.

Light protection of fiber-reinforced strip using aluminum foil for the direct splinting technique.

The critical point of the clinical procedure of direct splinting is the placement of the fiber-reinforced strip, which has to be well adapted to tooth surfaces. This article describes the use of aluminum foil to cover and protect the fiber-reinforced strip from light polymerization during the direct splinting technique. The advantage of using aluminum foil is to prevent premature curing of the fiber-reinforced strip, which would necessitate repetition of the procedure. With this technique, the strip is gradually exposed and adapted to the lingual/palatal surfaces of the teeth and light cured, allowing for ample working time.

[Bacteriological, clinical, and pharmacokinetic studies of perioperative antibiotic prophylaxis in head and neck surgery]. / Bakteriologische, klinische und pharmakokinetische Untersuchungen zur perioperativen Antibiotikaprophylaxe in der Kopf-Hals-Chirurgie.

The point of this study was to analyze the possible benefits of perioperative antibiotic prophylaxis in patients from whom oral cavity or throat tumors are removed. The criteria used to judge the efficacy of each treatment included the clinical course of the treatment, the bacterial colonization of the surgical area as well as the growth of bacteria during the postoperative phase. 50 patients were chosen and grouped according to their surgical treatment: laryngectomy (n = 20), partial laryngectomy (n = 22) or tongue, floor of the mouth, soft palate, gum or base of the tongue partial resection (n = 8). Within each surgical group, patients were randomly chosen for antibiotic prophylaxis; others constituted the untreated control group. The antibiotic prophylaxis consisted of 5 g Mezlocillin administered at the time of narcosis for 20 min followed by 0.5 g Metronidazol for 10 min. These medications were given in 8-hour intervals for three days following surgery. Investigation of the first 20 patients (prophylaxis group n = 7, control group n = 13) revealed that the combination of Mezlocillin and Metronidazol positively influenced post-operative recovery (no complications) while the patients without prophylactic antibiotic treatment suffered general or local complication leading to, in 10 cases, the necessity of postoperative therapy. On the basis of these results, the random grouping of the patients was ended and all 30 remaining patients were given the antibiotic prophylaxis. Regardless of antibiotic treatment, the great majority of microbes isolated from throat swabs and tracheal secretions were gram-negative, aerobic bacteria. A prerequisite for efficacious prophylaxis is that the antibiotics be applied before the operation, so that a sufficient concentration is present at the time of pharyngotomy. On the basis of pharmacokinetic investigations, administration of the antibiotic 30 min preoperatively fulfills this requirement. Further, our recommendation, based on our measurement of the spectrum of bacteria present and their growth is that the antibiotics be applied over a period of three days postoperatively. This recommendation is also based on the fact that some patients (those having undergone partial laryngectomy or tongue, floor of the mouth, base of the tongue partial resections) have suffered loss of the swallowing reflex so that there exists a continuous contamination of the surgical area with pathogens or facultative pathogens coming from the nasal or oral cavities.