Experimental identification of cancer driver alterations in the era of pan‐cancer genomics

Rapidly accumulating data from large‐scale cancer genomics studies have been generating important information about genes and their somatic alterations underlying cell transformation, cancer onset and tumor progression. However, these events are usually defined by using computational techniques, whereas the understanding of their actual functional roles and impact typically warrants validation by experimental means. Critical information has been obtained from targeted genetic perturbation (gene knockout) studies conducted in animals, yet these investigations are cost‐prohibitive and time‐consuming. In addition, the 3R principles (replacement, reduction, refinement) have been set in place to reduce animal use burden and are increasingly observed in many areas of biomedical research. Consequently, the focus has shifted to new designs of innovative cell‐based experimental models of cell immortalization and transformation in which the critical cancer driver events can be introduced by mutagenic insult and studied functionally, at the level of critical phenotypic readouts. From these efforts, primary cell‐based selective barrier‐bypass models of cell immortalization have emerged as an attractive system that allows studies of the functional relevance of acquired mutations as well as their role as candidate cancer driver events. In this review, we provide an overview of various experimental systems linking carcinogen exposure‐driven cell transformation with the study of cancer driver events. We further describe the advantages and disadvantages of the currently available cell‐based models while outlining future directions for in vitro modeling and functional testing of cancer driver events.     

Fatal neonatal nephrocutaneous syndrome in 18 Roma children with EGFR deficiency

Epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein with tyrosine-kinase signaling activity, involved in many cellular functions including cell growth and differentiation. Germ line loss-of-function mutations in EGFR lead to a severe neonatal skin disorder (Online Mendelian Inheritance in Man #131550). We report 18 premature Roma children from 16 families with birthweights ranging 440–1470 g and multisystem diseases due to the homozygous mutation c.1283G˃A (p.Gly428Asp) in EGFR. They presented with thin, translucent, fragile skin (14/15), skin desquamation (10/17), ichthyosis (9/17), recurrent skin infections and sepsis (9/12), nephromegaly (10/16) and congenital heart defects (7/17). Their prognosis was poor, and all died before the age of 6 months except one 13-year-old boy with a severe skin disorder, dentinogenesis imperfecta, Fanconi-like syndrome and secondary hyperaldosteronism. Management of ion and water imbalances and extremely demanding skin care may improve the unfavorable outcome of such patients.     

A Comparison of the Reactivating and Therapeutic Efficacy of Two Newly Developed Oximes (K727 and K733) with Oxime K203 and Trimedoxime in Tabun‐Poisoned Rats and Mice

The reactivating and therapeutic efficacy of three original bispyridinium oximes (K727, K733 and K203) and one currently available oxime (trimedoxime) was evaluated in tabun‐poisoned rats and mice. The oxime‐induced reactivation of tabun‐inhibited acetylcholinesterase was measured in diaphragm and brain of tabun‐poisoned rats. The results showed that the reactivating efficacy of two recently developed oximes (K727 and K733) does not achieve the level of the reactivation of tabun‐inhibited acetylcholinesterase induced by oxime K203 and trimedoxime. While all oximes studied were able to increase the activity of tabun‐inhibited acetylcholinesterase in diaphragm, oxime K733 was not able to reactivate tabun‐inhibited acetylcholinesterase in the brain. The therapeutic efficacy of all oximes studied roughly corresponds to their reactivating efficacy. While both recently developed oximes were able to reduce acute toxicity of tabun less than 1.5‐fold, another original oxime K203 and commonly used trimedoxime reduced the acute toxicity of tabun almost two times. In conclusion, the reactivating and therapeutic potency of both newly developed oximes does not prevail the effectiveness of oxime K203 and trimedoxime, and therefore, they are not suitable for their replacement of commonly used oximes for the antidotal treatment of acute tabun poisoning.     

Phytochemical Characterization,Antibacterial, Acetylcholinesterase Inhibitory and Cytotoxic Properties of Cryptostephanus vansonii,an Endemic Amaryllid

Cryptostephanus vansonii I. Verd., an endemic Amaryllidaceae species from Zimbabwe, was evaluated for its acetylcholinesterase (AChE) inhibitory and cytotoxicity properties using Ellman's colorimetric method and the tetrazolium‐based colorimetric assay against Vero monkey kidney cells, respectively. The plant extracts were also evaluated for their antibacterial activity against five bacteria. Furthermore, phytochemical profiles of the extracts were determined using ultra‐high performance liquid chromatography coupled with tandem mass spectrometry analysis. A plant part‐dependent AChE inhibitory activity was observed, in the order, root > rhizome > basal leaf > leaf. Overall, C. vansonii extracts exhibited better antibacterial activity against Gram‐negative compared with Gram‐positive bacteria. Cytotoxic effects were not detected in Vero monkey kidney cell lines suggesting the possible absence of toxophores in C. vansonii extracts. Similar to the trend in biological activity, a distinct plant part‐dependent variation in hydroxybenzoates, hydroxycinnamates and flavonoids was observed in the plant extracts. In addition, 5‐hydroxymetylfurfural and eucomic acid were detected in the different plant parts of C. vansonii. The results of the present study provide valuable AChE inhibition activity, toxicological and phytochemical profiles of C. vansonii. Further studies on isolation of bioactive compounds and their subsequent evaluation in other pharmacological and toxicological model systems are required. Copyright © 2017 John Wiley & Sons, Ltd.     

Effect of luting agents and reconstruction techniques on the fracture resistance of pre-fabricated post systems

Fracture resistance and fracture modes of endodontically treated maxillary central incisors restored with different post-and-core systems covered with all-ceramic copings were evaluated. Ten samples were prepared for each group. Groups 1, 2 and 3 consisted of tooth-coloured post-and-core, zirconia post (Cosmopost) with a composite core (Tetric Ceram), zirconia post (Cosmopost) with a custom made ceramic core (Cosmo Ingot), glass fibre-reinforced post (FRC Postec) with a composite core (Tetric Ceram), respectively. Group 4 consisted of a titanium post (ERpost) with a composite core (Tetric Ceram). The control group (group 5) consisted of root-filled incisors without posts. Tooth-coloured posts were cemented in the roots using Variolink-2, while titanium posts were cemented in the roots using Harvard cement. The all-ceramic copings were cemented using Variolink-2. Static load was applied to 2 mm below the incisal edge on the palatinal surface of each sample until they were fractured. Fracture data were obtained and statistically analysed with One-way anova and a Tukey's test. The results of the means and standard deviations of the fracture resistance during static loading were: 497.5 +/- 61.94 (1), 474.61 +/- 96.84 (2), 494.61 +/- 104.67 (3), 581.34 +/- 105.36 (4), 420.42 +/- 127.48 (5). There were statistically significant differences between groups 4 and 5. Glass fibre-reinforced posts and composite cores (group 3) showed the most catastrophic failure. Consequently, zirconia ceramic posts can be used in clinical practice.     

Influence of luting agent on the microleakage of all-ceramic crowns

PURPOSE: In this in-vitro study, microleakage of all-ceramic crowns was evaluated at enamel and dentin margins. MATERIALS AND METHODS: Forty maxillary central incisors were randomly divided into 4 groups (n = 10). While buccal and palatal margins were placed on enamel, mesial and distal margins were placed below the cementoenamel junction. In groups 1 to 3, IPS Empress 2 crowns were luted with Variolink 2/Syntac Classic (group 1), Bifix DC/Solobond Plus (group 2) and Calibra/Prime & Bond NT combinations (group 3), respectively. In the control group (group 4), porcelain-fused-to-metal crowns were luted with a zinc-phosphate cement. All specimens were subjected to 5000 thermocycles (at 5 degrees C to 55 degrees C; 30-s dwell time). After immersion in India ink for 48 h at 37 degrees C, the specimens were sectioned both buccolingually and mesiodistally. Each section was evaluated for microleakage under a stereomicroscope at 24X magnification. RESULTS: According to the Krukal-Wallis test, in all groups, there were significant differences in microleakage at the enamel margins (p = 0.001). Nevertheless, the margins finished in dentin showed no significant differences (p = 0.163). According to the Mann-Whitney U-test, statistically significant differences were observed in microleakage between groups 1 and 3 (p = 0.049), groups 1 and 4 (p = 0.001), groups 2 and 4 (p = 0.002), and between groups 3 and 4 (p = 0.045) at the enamel margin. In group 1, significantly greater microleakage was observed at the dentin margin compared to the enamel margin (p = 0.007). CONCLUSION: The adhesive luting technique demonstrated an excellent ability to minimize microleakage of all-ceramic crowns at the enamel margins. Water-based dentin bonding systems showed less microleakage than the water-free acetone-based dentin bonding system at the enamel margin.     

Steroid-sparing maintenance immunosuppression is safe and effective after simultaneous liver-kidney transplantation

Optimization of maintenance immunosuppression (mIS) regimens in the transplant recipient requires a balance between sufficient potency to prevent rejection and avoidance of excessive immunosuppression to prevent toxicities and complications. The optimal regimen after simultaneous liver-kidney (SLK) transplantation remains unclear, but small single-center reports have shown success with steroid-sparing regimens. We studied 4184 adult SLK recipients using the Scientific Registry of Transplant Recipients, from March 1, 2002, to February 28, 2017, on tacrolimus-based regimens at 1 year post-transplant. We determined the association between mIS regimen and mortality and graft failure using Cox proportional hazard models. The use of steroid-sparing regimens increased post-transplant, from 16.1% at discharge to 88.0% at 5 years. Using multi-level logistic regression modeling, we found center-level variation to be the major contributor to choice of mIS regimen (ICC 44.5%; 95% CI: 36.2%-53.0%). In multivariate analysis, use of a steroid-sparing regimen at 1 year was associated with a 21% decreased risk of mortality compared to steroid-containing regimens (aHR 0.79, P = .01) and 20% decreased risk of liver graft failure (aHR 0.80, P = .01), without differences in kidney graft loss risk (aHR 0.92, P = .6). Among SLK recipients, the use of a steroid-sparing regimen appears to be safe and effective without adverse effects on patient or graft survival.     

Treatment of gastrointestinal hemorrhage

Background We assessed the value of selective arteriography in the diagnosis and management of acute gastrointestinal hemorrhage. Methods We reviewed the records of 107 consecutive patients who had gastrointestinal hemorrhage and underwent selective arteriography between January 1992 and October 2003: 10 had upper gastrointestinal bleeding, 79 had lower gastrointestinal bleeding, and 18 had varicose bleeding with portal hypertension. Selective embolization was attempted in 15 patients to obtain hemostasis. Angiographic findings were reviewed and prospective reports were compared with the final diagnosis and outcome. Results Of 129 angiographic studies, 36 correctly revealed the bleeding site and 93 were negative. Extravasation was seen in 24 cases at the level of stomach (n = 2), duodenum (n = 1), small bowel (n = 5), or colon (n = 16). Indirect signs of bleeding sources were identified in 12 patients (stomach in one, small bowel in four, large bowel in four, liver in three). Transcatheter embolization induced definitive hemostasis in 11 of 15 patients (73%), namely in the stomach (n = 2), small bowel (n = 3), colon (n = 7), and liver (n = 3). Three patients required surgery after embolization. Conclusion Abdominal arteriography may localize gastrointestinal bleeding sources in approximately one-third of cases. Selective embolization may provide definitive hemostasis in most instances. P. Charbonnet and J. Toman contributed equally to this work.