Topical therapy for regression and melanoma prevention of congenital giant nevi.

Giant congenital melanocytic nevi are NRAS-driven proliferations that may cover up to 80% of the body surface. Their most dangerous consequence is progression to melanoma. This risk often triggers preemptive extensive surgical excisions in childhood, producing severe lifelong challenges. We have presented preclinical models, including multiple genetically engineered mice and xenografted human lesions, which enabled testing locally applied pharmacologic agents to avoid surgery. The murine models permitted the identification of proliferative versus senescent nevus phases and treatments targeting both. These nevi recapitulated the histologic and molecular features of human giant congenital nevi, including the risk of melanoma transformation. Cutaneously delivered MEK, PI3K, and c-KIT inhibitors or proinflammatory squaric acid dibutylester (SADBE) achieved major regressions. SADBE triggered innate immunity that ablated detectable nevocytes, fully prevented melanoma, and regressed human giant nevus xenografts. These findings reveal nevus mechanistic vulnerabilities and suggest opportunities for topical interventions that may alter the therapeutic options for children with congenital giant nevi.

Epitope spreading toward wild-type melanocyte-lineage antigens rescues suboptimal immune checkpoint blockade responses.

Although immune checkpoint inhibitors (ICIs), such as anti-programmed cell death protein-1 (PD-1), can deliver durable antitumor effects, most patients with cancer fail to respond. Recent studies suggest that ICI efficacy correlates with a higher load of tumor-specific neoantigens and development of vitiligo in patients with melanoma. Here, we report that patients with low melanoma neoantigen burdens who responded to ICI had tumors with higher expression of pigmentation-related genes. Moreover, expansion of peripheral blood CD8+ T cell populations specific for melanocyte antigens was observed only in patients who responded to anti-PD-1 therapy, suggesting that ICI can promote breakdown of tolerance toward tumor-lineage self-antigens. In a mouse model of poorly immunogenic melanomas, spreading of epitope recognition toward wild-type melanocyte antigens was associated with markedly improved anti-PD-1 efficacy in two independent approaches: introduction of neoantigens by ultraviolet (UV) B radiation mutagenesis or the therapeutic combination of ablative fractional photothermolysis plus imiquimod. Complete responses against UV mutation-bearing tumors after anti-PD-1 resulted in protection from subsequent engraftment of melanomas lacking any shared neoantigens, as well as pancreatic adenocarcinomas forcibly overexpressing melanocyte-lineage antigens. Our data demonstrate that somatic mutations are sufficient to provoke strong antitumor responses after checkpoint blockade, but long-term responses are not restricted to these putative neoantigens. Epitope spreading toward T cell recognition of wild-type tumor-lineage self-antigens represents a common pathway for successful response to ICI, which can be evoked in neoantigen-deficient tumors by combination therapy with ablative fractional photothermolysis and imiquimod.

Topical doxycycline foam 4% for prophylactic management of epidermal growth factor receptor inhibitor skin toxicity: an exploratory phase 2, randomized, double-blind clinical study.

PURPOSE: Acneiform rash, a common toxicity of epidermal growth factor receptor inhibitors (EGFRIs), can cause patient discomfort, warranting changes in treatment. This study investigated the safety, tolerability, and efficacy of a novel doxycycline foam, FDX104 4%, for managing EGFRI-related skin toxicity. METHODS: This was an exploratory phase 2, randomized, double-blind, placebo-controlled study. Subjects had metastatic colorectal cancer and were being treated with either cetuximab or panitumumab plus chemotherapy. Treatment (twice-daily topical FDX104 4% on one side of the face and vehicle foam on the other for 5 weeks) was initiated 7 ± 3 days prior to EGFRI therapy. Rash severity, safety, and tolerability were evaluated at 2 and 4 weeks after EGFRI start. RESULTS: The mean maximal rash grade was lower with FDX104 4% vs vehicle, and fewer subjects developed moderate-to-severe (grades 2-3) rash. On the Global Severity Score scale, a statistically significant difference favored FDX104 4% over vehicle (P = .047). Adverse events (AEs) (n = 68) occurred in 20 subjects; most were mild or moderate. The most common AEs were oral mucositis, nausea, and vomiting, common to chemotherapy and EGFRI treatment. Study-drug-related AEs were experienced by five subjects and consisted of mild, local skin reactions. No study-drug-related systemic side effects were reported. CONCLUSION: Twice-daily, topical administration of FDX104 4% as an adjunct to either cetuximab or panitumumab was safe and well tolerated, and appeared to prevent the onset of rash, especially severe rash. CLINICALTRIALS. GOV IDENTIFIER: Trial Registration NCT02239731.

Pathways in melanoma development.

This review aimed to summarize the current advances in melanoma research and their clinical relevance. Until recently, melanoma was considered an incurable disease. However, in recent years tremendous advances have been made in therapeutics. This is due to profound understanding of the molecular pathways involved in melanoma development. Although new targets are always emerging, understanding how to overcome resistance to current treatments is of great interest.

Temperature changes of one-piece implants during the setting of acrylic resin temporary crown. The effect of implant diameter. An in vitro study.

The purpose of this work is to evaluate changes in temperature of one-piece titanium implant surface during the setting of acrylic resin temporary crowns and to correlate thermal changes to implant diameter. Thirty-three one-piece implants (ARRP, Alpha-Biotec) were divided into 3 groups according to diameter size (G1=3 mm, G2=3.3 mm, G3=3.6 mm). Implants were mounted on an acrylic glass apparatus. Thermocouples were positioned at the most coronal thread. Lower incisor temporary polycarbonate crowns were filled with 80 µL of self-curing acrylic resin and positioned immediately on the implant abutment. Thermal changes of the implant surface were recorded continuously for 10 min. Data were statistically analyzed using one-way analysis of variance. The mean initial temperature (C0) of groups G1, G2 and G3 was similar (24.79±0.78ºC, 25.26±0.63ºC, 24.97±1.06ºC, respectively). The setting of the acrylic resin temporary crown resulted in a significant increase in the implant surface temperature of all groups. The mean thermal amplitude (ΔC) for groups G1, G2 and G3 were 6.79±1.02ºC, 6.61±0.94ºC, 6.65±1.26ºC, respectively. The mean time to maximum temperature (Tmax) for groups G1, G2 and G3 were 337.38±42.91 sec, 324.69±41.46 sec and 317.98±37.91 sec respectively (P>0.05). Direct application of auto-polymerizing resin to the titanium abutment of one-piece implants significantly increased the cervical implant surface temperature. Implant diameter did not influence the temperature changes.

Epigenomic reprogramming during pancreatic cancer progression links anabolic glucose metabolism to distant metastasis.

During the progression of pancreatic ductal adenocarcinoma (PDAC), heterogeneous subclonal populations emerge that drive primary tumor growth, regional spread, distant metastasis, and patient death. However, the genetics of metastases largely reflects that of the primary tumor in untreated patients, and PDAC driver mutations are shared by all subclones. This raises the possibility that an epigenetic process might operate during metastasis. Here we report large-scale reprogramming of chromatin modifications during the natural evolution of distant metastasis. Changes were targeted to thousands of large chromatin domains across the genome that collectively specified malignant traits, including euchromatin and large organized chromatin histone H3 lysine 9 (H3K9)-modified (LOCK) heterochromatin. Remarkably, distant metastases co-evolved a dependence on the oxidative branch of the pentose phosphate pathway (oxPPP), and oxPPP inhibition selectively reversed reprogrammed chromatin, malignant gene expression programs, and tumorigenesis. These findings suggest a model whereby linked metabolic-epigenetic programs are selected for enhanced tumorigenic fitness during the evolution of distant metastasis.

[Establishing adequate periimplant zone of keratinized mucosa to enhance periimplant health].

The presence of an adequate zone of keratinized mucosa (KM) for maintaining gingival health associated with natural teeth is usually described as the presence of 1-2 mm of attached gingiva (AG). Dental implants without KM measuring 2 mm or more, exposed to high levels of bacterial plaque, have shown higher scores of plaque accumulation, peri-implant 'mucosal inflammation', bleeding on probing, peri-implantitis, alveolar bone loss and recession. In a recent study the effects of KM width on peri-implant tissue health was assessed. Significant improvements in clinical and immunological parameters were noted after increasing KM width by the use of free gingival graft (FGG) surgery. A recent systematic review evaluated the efficacy of various techniques and biomaterials adopted in periimplant KM augmentation. Free gingival graft, connective tissue graft, acellular dermal matrix and collagen matrix were used for KM augmentation. Improvements in KM width were reported in all studies. A definitive conclusion could not be achieved owing to the lack of well-designed studies and appropriate methods of studying soft tissue. The establishment of universal surgical guidelines and measurement systems is imperative in the future. The present manuscript will describe the clinical use of FGG for establishing adequate periimplant zone of KM to enhance periimplant health.

Biomarker-guided screening of Juzen-taiho-to, an oriental herbal formulation for immunostimulation.

Juzen-taiho-to is an immunostimulatory herbal formulation that is clinically used in East Asia for cancer patients undergoing chemotherapy and radiation. The formulation stimulates various leukocytes, including T, B, and NK cells and macrophages. Although Juzen-taiho-to is known to contain numerous compounds with various pharmacological activities, it is not clear which compounds are responsible for the stimulation of individual cell types. Here, we conducted what we call "biomarker-guided screening" to purify compounds responsible for the macrophages stimulatory activity. To this end, gene expression was analyzed by a DNA array for macrophages treated with Juzen-taiho-to and DMSO (vehicle control), which identified intercellular adhesion molecule 1 as a biomarker of macrophage stimulation by Juzen-taiho-to. A quantitative reverse transcription polymerase chain reaction assay of intercellular adhesion molecule 1 was then used to guide the purification of active compounds. The screening resulted in the purification of a glycolipid mixture, containing ß-glucosylceramides. The glycolipid mixture potently stimulated intercellular adhesion molecule 1 expression in primary dendritic cells as well as in primary CD14+ (macrophages) cells. The identification of this glycolipid mixture opens up an opportunity for further studies to understand how plant-derived glycolipids stimulate macrophages and dendritic cells in a safe and effective manner as demonstrated by Juzen-taiho-to.

Immunostimulatory lipid nanoparticles from herbal medicine.

Reproducibility is an important issue in biological characterization of drug candidates and natural products. It is not uncommon to encounter cases in which supposedly the same sample exhibits very different biological activities. During our characterization of macrophage-stimulatory lipids from herbal medicine, it was found that the potency of these lipids could vary substantially from experiment to experiment. Further analysis of this reproducibility issue led to the discovery of solvent-dependent nanoparticle formation by these lipids. While larger nanoparticles (approximately 100 nm) of these lipids showed modest macrophage-stimulatory activity, smaller nanoparticles (<10 nm) of the same lipids exhibited substantially higher potency. Thus, the study revealed an unexpected link between nanoparticle formation and macrophage-stimulatory activity of plant lipids. Although nanoparticles have been extensively studied in the context of vehicles for drug delivery, our finding indicates that drugs themselves can form nanoassemblies, and their biological properties may be altered by the way they assemble.

[Animal experimentation, animal welfare and scientific research].

Hundreds of thousands of laboratory animals are being used every year for scientific experiments held in Israel, mostly mice, rats, rabbits, guinea pigs, and a few sheep, cattle, pigs, cats, dogs, and even a few dozen monkeys. In addition to the animals sacrificed to promote scientific research, millions of animals slain every year for other purposes such as meat and fine leather fashion industries. While opening a front against all is an impossible and perhaps an unjustified task, the state of Israel enacted the Animal Welfare (Animal Experimentation) Law (1994). The law aims to regulate scientific animal experiments and to find the appropriate balance between the need to continue to perform animal experiments for the advancement of research and medicine, and at the same time to avoid unnecessary trials and minimize animal suffering. Among other issues the law deals with the phylogenetic scale according to which experimental animals should be selected, experiments for teaching and practicing, and experiments for the cosmetic industry. This article discusses bioethics considerations in animal experiments as well as the criticism on the scientific validity of such experiments. It further deals with the vitality of animal studies and the moral and legal obligation to prevent suffering from laboratory animals.

[Soft tissue grafts for the treatment of mucogingival deformities and conditions on edentulous ridges].

Soft tissue grafts are commonly used to restore and/or improve mucogingival deformities and conditions around teeth or on edentulous ridges. This include vertical and/or horizontal ridge deficiency, lack of gingiva/keratinized tissue, decreased vestibular depth and abnormal color. The present article describes and illustrates different procedures using autologous soft tissue grafts including masticatory mucosal free graft, masticatory mucosal pedicle graft, subepithelial free connective tissue graft, and subepithelial dermal graft. The functional and esthetic advantages and limitations of each graft for each condition are discussed.

[Buccal bony exostoses induced by free gingival grafts].

Buccal Bony Exostoses (BBE) is a local benign osseous overgrowth continuous with the facial aspect of the jaw. Post operative BBE may be the result of dermal grafts used to restore the buccal vestibulum, of connective tissue graft placement, and of Free Gingival Grafts (FGG) procedures. In 46 patients in whom 72 FGG procedures were performed by the senior author (HT) over the past 12 years, BBE was clinically and radiographically diagnosed. In one case the tumor was surgically removed. The etiology, pathogenesis and frequency of BBE following FGG procedures was reviewed and discussed. We suggest that the BBE may develop owing to periosteal surgical trauma during FGG procedures, and suggest that this phenomenon receives further attention.

Rediscovery of natural products using genomic tools.

A screening methodology called 'genomic screening' was established to identify natural products that can regulate cellular gene expression. Application of genomic screening to Keishi-bukuryo-gan (KBG), a Japanese herbal medicine formulation, identified a previously unnoticed transcriptional effect by linoleic acid, a known KBG component. The approach opens up a possibility to develop cell-permeable molecular tools for functional genomics research and sets a stage to evaluate the potential of natural products for transcription therapies.

Gene expression profiling of intrinsic thermotolerance in Escherichia coli.

DNA microarrays were employed to compare gene expression in a thermotolerant, nalidixic acid-resistant mutant of Escherichia coli with that of the parental strain. When grown at 37 degrees C, up-regulated genes in the mutant included those coding for multiple antibiotic resistance proteins and enzymes for the degradation of small molecules, whereas among the down-regulated genes were those coding for fimbrial, flagellar, and outer membrane proteins as well as sigma 38. When the mutant grown at 42 degrees C was compared to the mutant grown at 37 degrees C, enhanced expression of several genes coding for flagellar proteins was detected. Reverse transcriptase-polymerase chain reaction analysis of selected genes confirmed results obtained with microarrays.

[Implants placement using a novel computerized tactile imaging navigation system].

An implant locating system has been developed. The system contains a stent that maps the jaw bone at the implantation site through the tissue, sends a signal to a computer with a CT scan of the jaw on it, and superimposes the position of the stent on the jaw so that a dental surgeon could angle the implant for placement without requiring to raise a flap to expose the bone. Using a novel tactile technology the system allows safe, accurate, and simple implant placement and design. The ILS software allows: 1. Importing of CT data. 2. Marking a dental arc on the computerized jaw image. 3. Planning of implant location and position on a 3-D view. 4. Affixation of the ILS to the jaw, followed by registration of the stent. 5. Navigated osteotomy and implant placement.

[Healing of osseous defects by guided bone regeneration using ribose cross linked collagen membranes].

The ultimate goal of periodontal therapy has long been the complete regeneration of the periodontal attachment apparatus. Guided Tissue Regeneration (GTR) and Guided Bone Regeneration (GBR) are two regenerative procedures which converted this goal from a dream to reality. In search of a biocompatible resorbable tissue barrier, collagen, being a natural protein and a weak antigen, has attracted much interest and became the focus of much intention during the 80's and the 90's. The understanding that cross linking of collagen with aldehyde sugars, especially ribose, produces collagen which is highly resistant to resorption in vivo led to the development of a "natural" Crossed-Linked Collagen Barrier (CB-SX). Animal and Human studies have shown that the newly developed membrane is biocompatible, remains intact in the tissues 6 months and more, and results in impressive guided tissue/bone regeneration. Spontaneous early exposure of the membrane is common but the healing potential of the resulted tissue dehiscence is favorable with no tendency for bacterial infection. The commercial version of the CB-SX is especially suitable for GBR procedures; it is highly recommended that the gingival flaps involved will properly be released, will lack tension, and be thoroughly sutured.

Rectal cancer: the impact of a colorectal unit on the preservation of the anal sphincter.

BACKGROUND: Mid- and lower rectum cancer is a technical challenge to the surgeon aiming to preserve the anal sphincter. The choice between abdominoperineal resection and anterior resection is often related to surgical skills. OBJECTIVES: To evaluate the role of a specialized colorectal unit in preserving the anal sphincter mechanism in the treatment of rectal cancer. METHODS: Between 1991 and 1996, 75 patients with rectal cancer up to 12 cm from the anal verge were operated at the Sheba Medical Center. Among them, 21 patients (group 1) underwent surgery in the colorectal unit and 54 patients (group 2) in the other two surgical departments. All patients had a complete preoperative investigation and were followed for 12-90 months. RESULTS: Background and tumor parameters were similar for both groups. In group 1, 20 patients (95%) had low anterior resection and 1 patient (5%) had abdominoperineal resection as compared to 20 patients (37%) and 34 patients (63%), respectively, in group 2 (P < 0.005). There was no statistical difference in the systemic recurrence rate. Local recurrence was more frequent in group 2 (P < 0.05). CONCLUSIONS: Special training in colorectal surgery enables the surgeon, in keeping with the principles of oncologic surgery, to preserve the anal sphincter mechanism in most patients with adenocarcinoma located in the mid- and lower third of the rectum.

Long-term follow-up and factors influencing success of silastic ring vertical gastroplasty.

BACKGROUND: Careful selection of patients for the operation plays a major role in long-term results of silastic ring vertical gastroplasty. The objective of the current study is to identify predictive factors for the objective (excess weight loss) and subjective (satisfaction) success of silastic ring vertical gastroplasty (SRVG) for morbid obesity. DESIGN: Retrospective cohort study. SETTING: University hospital. SUBJECTS: 300 patients of 450 who were operated on between 1984 and 1997. Follow-up time was 4.4 +/- 2.3 years. INTERVENTION: SRVG. STATISTICAL METHODS: Correlations, multi-linear regression model. RESULTS: Average BMI (body mass index) loss: 13.6 +/- 7.4 kg/m(2), average excess weight loss was 67.4 +/- 33.0%. Satisfaction rate: 81.3%. Correlation was found between objective parameters of successes and the satisfaction of the patient (p < 0.001). Excess weight loss was correlated to younger age (p < 0.005), pre-operative weight and BMI (p < 0.005, p < 0.01, respectively), and shorter follow-up (p < 0.001). Multiple linear regression model revealed that age and preoperative weight were independent variables and correlated to the excess weight loss after SRVG (R(2) = 0.303, p < 0.01; R(2) = 0.026, p < 0.05). Social support was correlated to satisfaction (p < 0.05). CONCLUSIONS: SRVG is an operation with high rates of objective and subjective success rate. Younger and heavier subjects will mostly enjoy SRVG in terms of excess weight loss. Patients who have social support have the most satisfactory emotional outcome.

Pre-operative localization of parathyroid adenoma by Tc-99m-sestamibi scintigraphy (MIBI)

The use of pre-operative imaging for localization of primary parathyroid adenoma may influence the duration and results of parathyroidectomy. The current study was aimed to evaluate the efficiency of localization of parathyroid adenoma by Tc-99m-sestamibi (MIBI) scintigraphy and compare the results with those achieved by the use of preoperative ultrasound. Seventy five patients, aged 25 to 83 years with primary hyperparathyroidism were operated due to primary adenoma in Rabin Medical Center from January 1995 to April 1997. Fifty of them had a preoperative MIBI scintigraphy and ultrasound for localization of parathyroid adenoma, while 25 had a preoperative ultrasound alone. Ultrasound identified correctly the adenoma in 84 percent of the cases, as compared to 96 percent identified by MIBI scintigraphy (p<0.01). MIBI scintigraphy shortened operation length from 120ñ20 min to 80ñ15 min (p<0.05) and reduced the number of frozen sections from 2.2ñ0.4 to 1.1ñ0.3 (p<0.001). MIBI scintigraphy is the most efficient modality for preoperative localization of parathyroid adenoma as compared to other imaging procedures, and can shorten operative time

Long-term follow-up in managing anaplastic astrocytoma by multimodality approach with surgery followed by postoperative radiotherapy and PCV-chemotherapy: phase II trial.

Overall survival and progression-free survival after 5 and 10 years of 31 patients with malignant glioma treated by a combination of surgery, postoperative radiotherapy, and chemotherapy with a PCV regimen (procarbazine, CCNU [lomustine] and vincristine) is described. Parameters were evaluated by age at diagnosis, gender, ethnic origin, pre- and postsurgery Karnofsky Performance Status (KPS) score, limit and amount of surgical resection, histopathologic type, number of chemotherapy courses, time between surgery and radiotherapy, response to combined therapy, and dosage and type of radiotherapy. Progression-free survival was 29% at 24 months and 22% at 60 and 120 months. Overall survival was 47%, 36%, and 36% after 24, 60, and 120 months, respectively. Favorable prognostic factors for survival in univariate analysis were pre- and postoperative KPS (> or =70; p = 0.015; p = 0.0025, respectively), age of patients (<40; p = 0.01), number of chemotherapy cycles (> or =6; p = 0.02), and radiation dose (> or =60 Gy; p = 0.0015). The only significant prognostic factors for overall survival in a stepwise multivariate analysis were irradiation dose (p = 0.0001), number of chemotherapy cycles (p = 0.001), and preoperative KPS (p = 0.05); for progression-free survival it was number of chemotherapy cycles (p = 0.004). Survival was not affected by excision size, radiation method, histopathologic type of tumor, gender, ethnic origin, or time lapsed between surgery and irradiation.