CRAF R391W is a melanoma driver oncogene.

Approximately 75% of melanomas have known driver oncogenic mutations in BRAF, NRAS, GNA11 or GNAQ, while the mutations providing constitutive oncogenic signaling in the remaining melanomas are not known. We established a melanoma cell line from a tumor with none of the common driver mutations. This cell line demonstrated a signaling profile similar to BRAF-mutants, but lacked sensitivity to the BRAF inhibitor vemurafenib. RNA-seq mutation data implicated CRAF R391W as the alternative driver mutation of this melanoma. CRAF R391W was homozygous and over expressed. These melanoma cells were highly sensitive to CRAF, but not BRAF knockdown. In reconstitution experiments, CRAF R391W, but not CRAF WT, transformed NIH3T3 cells in soft-agar colony formation assays, increased kinase activity in vitro, induced MAP kinase signaling and conferred vemurafenib resistance. MAP kinase inducing activity was dependent on CRAF dimerization. Thus, CRAF is a bona fide alternative oncogene for BRAF/NRAS/GNAQ/GNA11 wild type melanomas

Using the Hair Removal Laser in the Axillary Region and its Effect on Normal Microbial Flora

Introduction: The axillary hair removal laser is one of the most often used procedures to treat unwanted hairs in that region. Employing this technology can be helpful in decreasing the bromhidrosis.Methods: In the present research, a clinical trial study over the effect of the hair removal laser on normal microbial flora at the axillary region is presented. The intervention group consisted of 30 women referred to the dermatologic clinic for the purpose of removing axillary hair by the alexandrite 755 nm laser and the control group consisted of 30 women referred to the same clinic for any other reasons. Both groups were evaluated for the type of bacterial strains on the first visit and after three and six months.Results: The results showed that the sense of sweat smell improved by about 63% after the last laser session. The frequency of all bacterial strains decreased in the intervention group except Staphylococcus epidermidis which was significant. In the control group, there was no significant decrement in any bacterial strains and even the prevalence of more strains including Staphylococcus aureus and S. epidermidis increased. Counting the mean bacterial colon showed a slight decrement of the bacterial count following the laser.Conclusion: The use of laser radiation, even with the aim of hair removal, can alter the microbial flora, and it can be accompanied by the improvement of the smell of sweat. The effect of the laser on different bacterial strains is quite different, which can depend on the amount of energy, the wavelength, the characteristics of the area under the laser, and also the structural properties of the membrane of the microorganism itself

Effects of AKT inhibitor therapy in response and resistance to BRAF inhibition in melanoma

BackgroundThe clinical use of BRAF inhibitors for treatment of metastatic melanoma is limited by the development of drug resistance. In this study we investigated whether co-targeting the MAPK and the PI3K-AKT pathway can prevent emergence of resistance or provide additional growth inhibitory effects in vitro.MethodsAnti-tumor effects of the combination of the BRAF inhibitor (BRAFi) dabrafenib and GSK2141795B (AKTi) in a panel of 23 BRAF mutated melanoma cell lines were evaluated on growth inhibition by an ATP-based luminescent assay, on cell cycle and apoptosis by flow cytometry and on cell signaling by western blot. Moreover, we investigated the possibilities of delaying or reversing resistance or achieving further growth inhibition by combining AKTi with dabrafenib and/or the MEK inhibitor (MEKi) trametinib by using long term cultures.ResultsMore than 40% of the cell lines, including PTEN-/- and AKT mutants showed sensitivity to AKTi (IC50 < 1.5 μM). The combination of dabrafenib and AKTi synergistically potentiated growth inhibition in the majority of cell lines with IC50 > 5 nM dabrafenib. Combinatorial treatment induced apoptosis only in cell lines sensitive to AKTi. In long term cultures of a PTEN-/- cell line, combinatorial treatment with the MAPK inhibitors, dabrafenib and trametinib, and AKTi markedly delayed the emergence of drug resistance. Moreover, combining AKTi with the MAPK inhibitors from the beginning provided superior growth inhibitory effects compared to addition of AKTi upon development of resistance to MAPK inhibitors in this particular cell line.ConclusionsAKTi combined with BRAFi-based therapy may benefit patients with tumors harboring BRAF mutations and particularly PTEN deletions or AKT mutations

Antitumor activity of the ERK inhibitor SCH772984 [corrected] against BRAF mutant, NRAS mutant and wild-type melanoma.

BackgroundIn melanoma, dysregulation of the MAPK pathway, usually via BRAF(V600) or NRAS(Q61) somatic mutations, leads to constitutive ERK signaling. While BRAF inhibitors are initially effective for BRAF-mutant melanoma, no FDA-approved targeted therapies exist for BRAF-inhibitor-resistant BRAF(V600), NRAS mutant, or wild-type melanoma.MethodsThe 50% inhibitory concentration (IC50) of SCH772984, a novel inhibitor of ERK1/2, was determined in a panel of 50 melanoma cell lines. Effects on MAPK and AKT signaling by western blotting and cell cycle by flow cytometry were determined.ResultsSensitivity fell into three groups: sensitive, 50% inhibitory concentration (IC50) < 1 μM; intermediately sensitive, IC50 1-2 μM; and resistant, >2 μM. Fifteen of 21 (71%) BRAF mutants, including 4 with innate vemurafenib resistance, were sensitive to SCH772984. All three (100%) BRAF/NRAS double mutants, 11 of 14 (78%) NRAS mutants and 5 of 7 (71%) wild-type melanomas were sensitive. Among BRAF(V600) mutants with in vitro acquired resistance to vemurafenib, those with MAPK pathway reactivation as the mechanism of resistance were sensitive to SCH772984. SCH772984 caused G1 arrest and induced apoptosis.ConclusionsCombining vemurafenib and SCH722984 in BRAF mutant melanoma was synergistic in a majority of cell lines and significantly delayed the onset of acquired resistance in long term in vitro assays. Therefore, SCH772984 may be clinically applicable as a treatment for non-BRAF mutant melanoma or in BRAF-mutant melanoma with innate or acquired resistance, alone or in combination with BRAF inhibitors

JUN dependency in distinct early and late BRAF inhibition adaptation states of melanoma.

A prominent mechanism of acquired resistance to BRAF inhibitors in BRAF (V600) -mutant melanoma is associated with the upregulation of receptor tyrosine kinases. Evidences suggested that this resistance mechanism is part of a more complex cellular adaptation process. Using an integrative strategy, we found this mechanism to invoke extensive transcriptomic, (phospho-) proteomic and phenotypic alterations that accompany a cellular transition to a de-differentiated, mesenchymal and invasive state. Even short-term BRAF-inhibitor exposure leads to an early adaptive, differentiation state change-characterized by a slow-cycling, persistent state. The early persistent state is distinct from the late proliferative, resistant state. However, both differentiation states share common signaling alterations including JUN upregulation. Motivated by the similarities, we found that co-targeting of BRAF and JUN is synergistic in killing fully resistant cells; and when used up-front, co-targeting substantially impairs the formation of the persistent subpopulation. We confirmed that JUN upregulation is a common response to BRAF inhibitor treatment in clinically treated patient tumors. Our findings demonstrate that events shared between early- and late-adaptation states provide candidate up-front co-treatment targets

Lactate, a Neglected Factor for Diabetes and Cancer Interaction

Increasing body of evidence suggests that there exists a connection between diabetes and cancer. Nevertheless, to date, the potential reasons for this association are still poorly understood and currently there is no clinical evidence available to direct the proper management of patients presenting with these two diseases concomitantly. Both cancer and diabetes have been associated with abnormal lactate metabolism and high level of lactate production is the key biological property of these diseases. Conversely, high lactate contribute to a higher insulin resistant status and a more malignant phenotype of cancer cells, promoting diabetes and cancer development and progression. In view of associations between diabetes and cancers, the role of high lactate production in diabetes and cancer interaction should not be neglected. Here, we review the available evidence of lactate's role in different biological characteristics of diabetes and cancer and interactive relationship between them. Understanding the molecular mechanisms behind metabolic remodeling of diabetes- and cancer-related signaling would endow novel preventive and therapeutic approaches for diabetes and cancer treatment

Therapeutic effects of <i>Rosa Canina, Urtica Dioica</i> and <i>Tanacetum Vulgare</i> Herbal Combination in Treatment of Tinnitus Symptoms; A Double-blind Randomized Clinical Trial

BackgroundTinnitus is defined as the perception of sound in the ear or head in the absence of an external stimulus for which we have no definite treatment. Neurotec® is a medication of herbal origin with IFDA approval. Previous studies showed the neuroprotective effect of Neurotec®. In this study we evaluated the effectiveness of Neurotec in improving tinnitus symptoms.MethodsThis double-blind randomized clinical trial was performed on patients with tinnitus. Patients received Neurotec 100 mg capsules (BID) or placebo for three months. Pure tone audiometry (PTA) was measured at 0.5, 1, 2, 4 and 6 KHz frequencies. Using a Tinnitus Handicap Inventory (THI) questionnaire, tinnitus loudness, daily annoyance, daily life or sleep disturbance, daily perception and mood alteration were evaluated.ResultsFinally, 103 (69 male and 34 female) patients with a mean age of 51.33±13.91 years were analyzed. There was no significant difference between the intervention (n=53) and the control group (n=50) regarding baseline symptoms before and one month after the intervention (P>0.05). While, they were significantly different three months after the intervention (P0.05). The mean pure tone air conduction was not significantly different between the two groups before and three months after the intervention at 6 kHz (P>0.05).ConclusionA three-month treatment with Neurotec Capsules beside patient education can effectively control symptoms of patients with tinnitus