The impact of SF3B1 mutations in CLL on the DNA-damage response.

Mutations or deletions in TP53 or ATM are well-known determinants of poor prognosis in chronic lymphocytic leukemia (CLL), but only account for approximately 40% of chemo-resistant patients. Genome-wide sequencing has uncovered novel mutations in the splicing factor sf3b1, that were in part associated with ATM aberrations, suggesting functional synergy. We first performed detailed genetic analyses in a CLL cohort (n=110) containing ATM, SF3B1 and TP53 gene defects. Next, we applied a newly developed multiplex assay for p53/ATM target gene induction and measured apoptotic responses to DNA damage. Interestingly, SF3B1 mutated samples without concurrent ATM and TP53 aberrations (sole SF3B1) displayed partially defective ATM/p53 transcriptional and apoptotic responses to various DNA-damaging regimens. In contrast, NOTCH1 or K/N-RAS mutated CLL displayed normal responses in p53/ATM target gene induction and apoptosis. In sole SF3B1 mutated cases, ATM kinase function remained intact, and γH2AX formation, a marker for DNA damage, was increased at baseline and upon irradiation. Our data demonstrate that single mutations in sf3b1 are associated with increased DNA damage and/or an aberrant response to DNA damage. Together, our observations may offer an explanation for the poor prognosis associated with SF3B1 mutations.

Interactions between the stratum corneum and topically applied products: regulatory, instrumental and formulation issues with focus on moisturizers.

Virtually everyone in Europe will use at least one cosmetic product every day. The extensive use of cosmetics and results from measurements of quality of life in patients with skin diseases demonstrate the importance of a healthy skin. The skin is not only a barrier against desiccation and intrusion of harmful materials, but also an organ of social communication, where dry, scaly, rough stratum corneum is unappealing to touch, inducing anxiety and depression. Knowledge about the skin biochemistry and the use of noninvasive instruments facilitate the development of topical products and quantification of their effects. The presentation of the products and mode of action determine the regulatory demands and the approval process, as they can fall into different regulatory entities, such as cosmetics, medicinal products, medical devices and as other chemical products. The majority of the topical products on the market are regulated as cosmetics. For example, facial skin care and daily moisturizing routines are frequently used. However, despite visible relief of dryness symptoms, some products are reported to result in deterioration of the skin barrier function. New clinical outcomes show important clinical differences between formulations and the relapse of eczema. In a worst case scenario, treatment with a moisturizing cream may increase the risks of eczema and asthma. In the present overview, product presentations and mode of actions are reflected against the regulatory demands in Europe. The regulations are continuously revisited and new guidelines are being implemented, such as the new cosmetic regulation with advice on testing and responsible marketing.

Effects of water activity and low molecular weight humectants on skin permeability and hydration dynamics - a double-blind, randomized and controlled study.

OBJECTIVES: The mammalian skin is a barrier that effectively separates the water-rich interior of the body from the normally dryer exterior. Changes in the external conditions, for example ambient humidity, have been shown to affect the skin barrier properties. The prime objective of this study was to evaluate the effect of water activity of a topical formulation on skin hydration and permeability. A second objective was to gain more understanding on how two commonly used humectants, urea and glycerol, affect skin barrier function in vivo. METHODS: Simple aqueous formulations were applied under occlusion to the volar forearm of healthy volunteers. Following 4-h exposure, skin water loss (by transepidermal water loss measurements), skin hydration (by Corneometry) and skin permeability (by time to vasodilation due to benzyl nicotinate exposure) were monitored. RESULTS: The results demonstrate that a relatively small change in the water activity of a topical formulation is sufficient to induce considerable effects on stratum corneum hydration and permeability to exogenous substances. Exposing the skin to high water activity leads to increased skin hydration and also increased permeability. Furthermore, urea and glycerol promote skin hydration and permeability even at reduced water activity of the applied formulation. CONCLUSION: These results highlight the importance of considering the water activity in topically applied formulations and the potential benefit of using humectants. The results may impact formulation optimization in how to facilitate skin hydration and to modify skin permeability by temporarily open and close the skin barrier.

The effect of a corticosteroid cream and a barrier-strengthening moisturizer in hand eczema. A double-blind, randomized, prospective, parallel group clinical trial.

BACKGROUND: Hand eczema is a common and persistent disease with a relapsing course. Clinical data suggest that once daily treatment with corticosteroids is just as effective as twice daily treatment. OBJECTIVES: The aim of this study was to compare once and twice daily applications of a strong corticosteroid cream in addition to maintenance therapy with a moisturizer in patients with a recent relapse of hand eczema. METHODS: The study was a parallel, double-blind, randomized, clinical trial on 44 patients. Twice daily application of a strong corticosteroid cream (betamethasone valerate 0.1%) was compared with once daily application, where a urea-containing moisturizer was substituted for the corticosteroid cream in the morning. The investigator scored the presence of eczema and the patients judged the health-related quality of life (HRQoL) using the Dermatology Life Quality Index (DLQI), which measures how much the patient's skin problem has affected his/her life over the past week. The patients also judged the severity of their eczema daily on a visual analogue scale. RESULTS: Both groups improved in terms of eczema and DLQI. However, the clinical scoring demonstrated that once daily application of corticosteroid was superior to twice daily application in diminishing eczema, especially in the group of patients with lower eczema scores at inclusion. CONCLUSIONS: Twice daily use of corticosteroids was not superior to once daily use in treating eczema. On the contrary, the clinical assessment showed a larger benefit from once daily treatment compared with twice daily, especially in the group of patients with a moderate eczema at inclusion.

Sunscreen use: controversies, challenges and regulatory aspects.

Mismatches between skin pigmentation and modern lifestyle continue to challenge our naked skin. One of our responses to these challenges is the development and use of sunscreens. The management of sunscreens has to balance their protective effect against erythema, photocarcinogenesis and photoageing owing to the potential toxicity of the ultraviolet (UV) filters for humans and the environment. The protection against UV radiation offered by sunscreens was recently standardized in the European Union (EU) based on international harmonization of measurement techniques. Four different categories of sun protection have been implemented along with recommendations on how to use sunscreen products in order to obtain the labelled protection. The UV filters in sunscreens have long been authorized for use by the EU authority on the basis of data from studies on acute toxicity, subchronic and chronic toxicity, reproductive toxicity, genotoxicity, photogenotoxicity, carcinogenicity, irritation, sensitization, phototoxicity and photosensitization as well as on environmental aspects. New challenges with respect to the safety of UV filters have arisen from the banning of animal experiments for the development of cosmetics. Future debates on sunscreens are likely to focus on nanoparticles and environmental issues, along with motivation campaigns to persuade consumers to protect their skin. However, more efficient sunscreen use will also continue to raise questions on the benefit in preventing vitamin D synthesis in the skin induced by sunlight.

Treatment with a barrier-strengthening moisturizing cream delays relapse of atopic dermatitis: a prospective and randomized controlled clinical trial.

BACKGROUND: Standard treatment of atopic dermatitis (AD) is based on topical glucocorticosteroids or calcineurin inhibitors to treat flares combined with moisturizer treatment to alleviate dry skin symptoms. Patients with AD have an abnormal skin barrier function, and strategies for reducing the risks for eczema would be to repair the barrier or prevent barrier dysfunction. OBJECTIVES: The objective of this study was to explore the time to relapse of eczema during a 26-week maintenance treatment with a urea containing moisturizer compared to no treatment (neither medical nor non-medicated preparations) after successful clearing of atopic lesions. The moisturizer has previously been shown to improve skin barrier function. METHODS: Patients applied betamethasone valerate (0.1%) on eczematous lesions during a 3-week period. Those with cleared eczema entered a 26-week maintenance phase, applying the moisturizer or left the previously affected area untreated. Upon eczema relapse, patients were instructed to contact the clinic and to have the relapse confirmed by the investigator. RESULTS: Fifty-five patients entered the study and 44 patients were included in the maintenance phase (22 using moisturizer twice daily and 22 using no treatment). Median time to relapse for patients treated with moisturizer was > 180 days (duration of the study) compared with 30 days for the no-treatment group. Sixty-eight per cent of the patients treated with the moisturizer and 32% of the untreated patients remained free from eczema during the observation period. CONCLUSIONS: Maintenance treatment with a barrier-improving urea moisturizer on previous eczematous areas reduced the risk of relapse to approximately one third of that of no treatment.

Enhancement of bioavailability by lowering of fat content in topical formulations.

BACKGROUND: The cosmetic properties of topical formulations are important parameters for the adherence to treatment, where modern oil-in-water emulsions are considered more acceptable compared with ointments. After application of an emulsion to the skin, the concentration of active ingredients in the formulation residue on the skin will increase, due to evaporation of volatile ingredients. OBJECTIVES: The aim of the present study was to investigate the effect of changes in vehicle fatty content on the skin penetration of two active ingredients: benzyl nicotinate (BN) and betamethasone valerate (BV). METHODS: Formulations containing 0.5% BN and 0.3% BV in vehicles with different lipid content (10-80%) were applied in a randomized and double-blind manner to the forearm of healthy volunteers. The changes in skin colour (erythema and blanching) were then monitored visually and with a new noninvasive instrument. RESULTS: The BN formulation containing 10% fat induced erythema more rapidly and with higher intensity than the formulations with higher fat content. Increased efficacy was also observed from the low-fat content formulation of BV, which gave more blanching than the formulations with high fat content. CONCLUSIONS: The rate of penetration of the active ingredients was inversely related to the lipid content, i.e. simple changes of the cosmetic properties by modifications of the lipid content may affect the efficacy of a formulation.

Artificial reduction in transepidermal water loss improves skin barrier function.

BACKGROUND: Artificial reduction of abnormal transepidermal water loss (TEWL) is considered to improve skin diseases associated with a defective barrier function. Treatment of the skin with moisturizers is also known to influence skin barrier function. Whether or not differences in occlusion between creams contribute to their effects on the skin barrier function is unknown. OBJECTIVES: To investigate the long-term effects of a semipermeable membrane on the skin barrier function in normal skin. In addition, the occlusive properties of two creams were studied. METHODS: The study was randomized, controlled and evaluator-blind using measurement of TEWL and skin susceptibility to sodium lauryl sulphate as indicators of skin barrier function. RESULTS: Coating of the skin with a silicone membrane for 23 h per day for 3 weeks improved skin barrier function, whereas no significant changes were found after using the membrane for 8 h per day. CONCLUSIONS: Differences between creams in terms of their effect on skin barrier function cannot be solely explained by their occlusive properties.

Changes in skin barrier function following long-term treatment with moisturizers, a randomized controlled trial.

BACKGROUND: Moisturizers are commonly used by patients with dry skin conditions as well as people with healthy skin. Previous studies on short-term treatment have shown that moisturizers can weaken or strengthen skin barrier function and also influence skin barrier recovery. However, knowledge of the effects on skin barrier function of long-term treatment with moisturizers is still scarce. OBJECTIVES: To investigate the impact of long-term treatment with moisturizers on the barrier function of normal skin, as measured by transepidermal water loss (TEWL) and susceptibility to an irritant, and to relate those effects to the composition of the designed experimental moisturizers. METHODS: Volunteers (n = 78) were randomized into five groups. Each group treated one volar forearm for 7 weeks with one of the following preparations: (i) one of three simplified creams, containing only a few ingredients in order to minimize the complexity of the system; (ii) a lipid-free gel; (iii) one ordinary cream, containing 5% urea, which has previously been shown to decrease TEWL. The lipids in the simplified creams were either hydrocarbons or vegetable triglyceride oil, and one of them also contained 5% urea. After 7 weeks, treated and control forearms were exposed for 24 h to sodium lauryl sulfate (SLS) using a patch test. TEWL, blood flow and skin capacitance of both SLS-exposed and undamaged skin were evaluated 24 h after removal of patches. Additionally, a 24-h irritancy patch test of all test preparations was performed on 11 volunteers in order to check their possible acute irritancy potential. RESULTS: Changes were found in the barrier function of normal skin after 7 weeks of treatment with the test preparations. The simplified creams and the lipid-free gel increased TEWL and skin response to SLS, while the ordinary cream had the opposite effect. One of the simplified creams also decreased skin capacitance. All test preparations were shown to be nonirritant, both by short-term irritancy patch test and by measurement of blood flow after long-term treatment. CONCLUSIONS: Moisturizers influence the skin barrier function of normal skin, as measured by TEWL and susceptibility to SLS. Moreover, the effect on skin barrier function is determined by the composition of the moisturizer. The ingredients which influence the skin barrier function need to be identified, and the mechanism clarified at the molecular level.

Increasing quality of life by improving the quality of skin in patients with atopic dermatitis.

Atopic dermatitis is a chronic relapsing inflammatory skin disease which usually starts during the first years of life. In patients with the disease, the quality of skin is severely affected, and this is closely linked to a reduced quality of life. An increasing prevalence of the disease has also been observed during recent years, which has been attributed to potential provocation factors in the environment. The environmental influence of the disease is complex, but the role of stratum corneum as a biosensor regulating the response to a variety of insults has been suggested as one crucial factor. Therefore, our daily hygiene and treatment of dryness are necessary measures to improve the quality of life and possibly reduce the frequency of the disease. Soaps as well as moisturizers show important differences in their impact on barrier function.

A double-blind and controlled study on the influence of the vehicle on the skin susceptibility to stinging from lactic acid.

Synopsis For patients with skin diseases, the process of treating the skin with topical medications adds to the burden of having the disease. Inconvenient skin reactions can make the treatment troublesome and lower the compliance. Moreover, epidemiological surveys indicate that 50% or more of female consumers believe they have sensitive skin. In the present study, the influence of the vehicle on the adverse skin reaction to lactic acid was judged by the test subjects after application of the test formulations to the facial skin. The results showed a water-in-oil (w/o) emulsion to induce less stinging than an ordinary oil-in-water (o/w) emulsion. Increasing the mineral oil content in the o/w emulsion from 10% to 50% tended (P = 0.077) to decrease the stinging potential of the formulation. An o/w emulsion free from lactic acid but with pH adjusted to 3 using hydrochloric acid induced significantly less stinging than the corresponding lactic acid formulation at pH 3. In conclusion, the present study gives new insights into the influence of vehicle on the stinging capacity of lactic acid, which may be related to its possible penetration via appendages. Hence, encapsulation of the stinging substances in the inner water phase of an emulsion may be a possible option to reduce adverse skin reactions and to increase compliance to water-soluble substances.

The clinical benefit of moisturizers.

UNLABELLED: Moisturizing creams marketed to consumers often contain trendy ingredients and are accompanied by exciting names and attractive claims. Moisturizers are also an important part of the dermatologist's armamentarium to treat dry skin conditions and maintain healthy skin. The products can be regarded as cosmetics, but may also be regulated as medicinal products if they are marketed against dry skin diseases, such as atopic dermatitis and ichthyosis. When moisturizers are used on the so-called dry skin, many distinct disorders that manifest themselves with the generally recognized symptoms of dryness are treated. Dryness is not a single entity, but is characterized by differences in chemistry and morphology in the epidermis depending on the internal and external stressors of the skin. Patients and the society expect dermatologists and pharmacists to be able to recommend treatment for various dry skin conditions upon evidence-based medicine. LEARNING OBJECTIVE: Upon completing this paper, the reader should be aware of different types of moisturizers and their major constituents. Furthermore, s/he will know more about the relief of dryness symptoms and the functional changes of the skin induced by moisturizers.

Irritation potential of bath and shower oils before and after use: a double-blind randomized study.

BACKGROUND: Difficulties in avoiding weak irritants may contribute to chronic contact dermatitis. A large variety of shower and bath oils are claimed to be suitable for use on dry skin because of their mildness and because they deposit a protective oil film on the skin. OBJECTIVES: The aim of the present study was to investigate possible differences in the irritation potential of eight shower or bath oils and to investigate whether surfactant residues may form a reservoir of irritant substance on the skin. PATIENTS AND METHODS: The study was double-blind and randomized using healthy human volunteers. The inherent capacity of the products to induce irritation was determined using conventional patch test techniques. Detection of potentially irritant residues was done by occlusion of the treated and rinsed skin area, followed by evaluation of the biological response. Instrumental measurements of transepidermal water loss and superficial skin blood flow served as indicators of the injurious effects of the products. RESULTS AND CONCLUSIONS: The results showed large differences between the products in irritant potential. Some did not irritate skin more than water, whereas others demonstrated considerably damaging effects. Moreover, the study proved the presence of barrier-impairing residues on the skin after rinsing with water. Thus, instead of protecting the skin, some formulations may induce subclinical injuries and delay skin barrier function recovery with prolonged risk for patients with eczema.

Do moisturizers work?

Moisturizers are used on large body surfaces to maintain the smoothness of the skin and to break the dry-skin cycle. Many healthcare professionals and patients overlook the importance of moisturizers and do not consider them to be 'active' treatments. However, evidence from clinical and experimental studies shows that moisturizers enhance both the smoothness and hydration of skin. Different moisturizers have different ingredients, and each may have a different mode of action. Some smooth the skin, others affect barrier function. Some enhance barrier function in both diseased and normal skin. Others impair barrier function in both diseased and normal skin. Defective barrier function may trigger the development of eczema. The composition of a particular moisturizer should reflect its desired therapeutic effect, i.e. a moisturizer to diminish dryness may need different ingredients from those required to improve barrier function. The content of excipients, such as emulsifiers, chelating agents and antioxidants, may have greater impact than is commonly believed. Greater tailoring of moisturizers will improve their efficacy. Confidence in the therapeutic effects of moisturizers will be enhanced by well-designed randomized controlled trials.

Mascaras may cause irritant contact dermatitis.

The majority of adverse effects of cosmetics have been attributed to soaps in Dutch and English studies, but to eye makeup in a recent Swedish study. The reactions may be caused by irritants or by sensitizing substances. The aim of the present study was to evaluate the irritation potential of commercially available mascaras. The mascaras were exposed to the skin in aluminium chambers. The skin reaction was evaluated using both visual assessments of erythema and non-invasive measurements of the skin reaction. Seven mascaras were tested on 15 healthy individuals in a randomized and blinded fashion. Two of the seven tested mascaras induced pronounced skin inflammation, when applied to normal skin under occlusion. These two mascaras were based on volatile petroleum distillate, in contrast to the other five mascaras that were conventional emulsions with stearate as the main emulsifier. The findings suggest that solvent-based mascaras might induce contact dermatitis due to its content of irritating substances.

Instrumental and dermatologist evaluation of the effect of glycerine and urea on dry skin in atopic dermatitis.

BACKGROUND/AIMS: Moisturising creams are useful treatment adjuncts in inflammatory dermatoses and have beneficial effects in the treatment of dry, scaly skin. The effects on dryness and skin permeability of a new moisturising cream with 20% glycerine was compared with its placebo and with a medicinally authorised cream with 4% urea (combined with 4% sodium chloride) in the treatment of dry skin. METHODS: Patients (n=109) with atopic dermatitis were treated for 30 days with a moisturiser in a randomised, parallel and double-blind fashion. Transepidermal water loss (TEWL) and skin capacitance were assessed instrumentally, and changes in the dryness of the skin were assessed by the dermatologist. RESULTS: No difference in TEWL was found between glycerine treatment and its placebo, whereas a lower value was found in the urea-treated area compared to the glycerine-treated area. No difference in skin capacitance was found. The clinical assessment of dryness showed urea to be superior to glycerine in treating the condition. CONCLUSIONS: Moisturising creams are different, not only with respect to composition but also with respect to their influence on skin as a barrier to water in patients with atopic dermatitis.

Strategy to decrease the risk of adverse effects of fragrance ingredients in cosmetic products.

In spite of extensive self-regulation of the fragrance industry, fragrance ingredients are still major causes of allergic contact dermatitis. There are indications that the problem is increasing in some countries, and that many nonregulated compounds are involved in the development of allergies. The use of essential oils in fragrance compounds might add both allergenic and carcinogenic compounds to a product and the exact composition of such ingredients is difficult to control. Herein, we propose a simple strategy to decrease the risk of adverse effects of fragrance ingredients in cosmetic products. This strategy consists of four major steps: (1) limit the concentration of fragrance compound in the products, (2) follow legislation and guidelines, (3) limit the concentration of a number of well-known sensitizing fragrance chemicals, and (4) limit the concentration of essential oils and materials with unknown composition. The strategy is discussed as an alternative to animal testing and in relation to other more resource-demanding approaches to the same problem.

The influence of a cream containing 20% glycerin and its vehicle on skin barrier properties.

Glycerin is widely used in cosmetics and well as in pharmaceutical formulations, mainly as humectant. In vitro studies have shown glycerin to prevent crystallization of stratum corneum model lipid mixture at low room humidity. Whether this may affect the skin barrier function during repeated application of glycerin in a cream base to normal skin is not known. Therefore, the influence of a cream containing 20% glycerin was compared with its placebo cream in a bilateral, double-blind study on 17 healthy volunteers. The effect was evaluated as influence on hydration with a corneometer and on skin barrier function. Skin barrier function was assessed as permeability to water with an evaporimeter (transepidermal water loss; TEWL) and as sensitivity to an irritating surfactant by measuring the biological response (measured as TEWL and skin blood flow). Ten days treatment of normal skin with 20% glycerin significantly increased skin corneometer values, indicating an increased hydration. However, our study failed to show an influence of glycerin on human skin, in terms of TEWL and skin sensitivity to SLS-induced irritation.

Content of fragrance mix ingredients and customer complaints of cosmetic products.

BACKGROUND: In relation to the wide use of cosmetics, serious adverse effects are rare. Occasionally, unwanted effects such as contact dermatitis are reported. Allergic reactions to cosmetics are often caused by fragrances. OBJECTIVE: The aim was to investigate the content of fragrance mix (FM) ingredients in cosmetic products of the brand ACO HUD (Stockholm, Sweden) and the frequency of customer skin complaints about fragranced and unfragranced products over 4.5 years. METHOD: Content of FM ingredients in the fragrances used was both analyzed and requested from the suppliers. Customer complaints were those reported to the company. RESULTS: Between 1 and 7 of FM ingredients were present in levels of less than 0.1 to 770 ppm. The ingredients, in order of frequency, were geraniol, eugenol, hydroxycitronellal, alpha-amyl cinnamic aldehyde, isoeugenol, cinnamic alcohol, and oak moss. Cinnamic aldehyde was not found. No significant difference was found either between the frequency of complaints about products with and without fragrance (P = .21) or in a paired comparison of 17 formulas marketed with and without fragrance (P = .24). CONCLUSION: The study suggests that the investigated fragranced products had a low content of FM ingredients, which might explain the absence of a higher frequency of adversities. Furthermore, it appears that under such circumstances fragrances may be used without introducing an increased rate of spontaneous complaints of skin reactions.

Long-term cultured IL-2-dependent T cell lines demonstrate p16(INK4a) overexpression, normal pRb/p53, and upregulation of cyclins E or D2.

Acquisition of an immortal phenotype by circumvention of the normal senescence program can be an important step in tumor development and progression. The regulation of life-span checkpoints is complex and abrogation of these processes can occur at different levels. To better understand these mechanisms in long-term cultured lymphocytes we have characterized two human long-term cultured IL-2-dependent T cell lines regarding telomere length, telomerase activity, and the expression of selected cell cycle regulators (pRb, p53, cyclin E, cyclin D1, cyclin D2, cyclin D3, cdk4, p16(INK4a), p21(WAF1), p27(KIP1), c-myc, bcl-2, and NPAT). We compared these cell lines with a primary T lymphoblast population with a limited life span from the same donor. Both T cell lines with extraordinary growth capacity showed telomere length stabilization, high telomerase activity and demonstrated wild-type pattern of pRb and p53 but strong p16(INK4a) protein expression. The growth inhibitory activity of p16(INK4a) seemed to be abrogated by enhanced expression of cyclin D2, cdk4, and c-myc in one T cell line and overexpression of cyclin E in the second T cell line.