Interleukin-12 (IL-12) is a heterodimeric cytokine originally defined by its ability to induce the maturation of cytolytic lymphocytes and by its capacity to effectively synergize with IL-2 in the induction of cytolytic activity. Recent studies in mice have demonstrated the ability of IL-12 to cause tumor regression and stimulate long-term antitumor immunity in treated animals. To examine the antitumor effect of direct gene transfer of IL-12 into tumors, we have developed retroviral vectors that coordinately express both subunits of IL-12. An MFG-based retroviral vector was used to generate a recombinant retrovirus in which a long terminal repeat (LTR)-driven polycistronic transcript encodes both subunits of human IL-12: hp35 and hp40 cDNAs are linked and coexpressed using the internal ribosome entry site (IRES) from the encephalomyocarditis virus (DFG-hIL-12). In addition, two IRES sequences were used to express both subunits of IL-12 and a neomycin resistance (neoR) selectable marker gene from the same polycistronic message (TFG-hIL-12). The amphotropic DFG-hIL-12 and TFG-hIL-12 viruses were used to infect both human and murine cell lines as well as primary tumor cultures. The production of human IL-12 by the nonselected, infected cells was measured in both a PHA blast proliferation bioassay and an ELISA and ranged from 15 to 40 ng/10(6) cells per 24 hr. Following G418 selection of TFG-hIL-12-infected cells, the level of expression of IL-12 was significantly higher (up to 120 ng/10(6) cells per 24 hr). The IL-12 protein secreted by the infected cells exhibited all of the biologic activities of recombinant hIL-12: proliferation of activated natural killer (NK) and T cells, stimulation of interferon-gamma (IFN-gamma) induction by NK and T cells, and enhancement of lymphokine-activated killer (LAK) activity. These retroviral vectors expressing human IL-12 should be useful in evaluating the biological properties of IL-12 as well as for use in clinical trials for gene therapy of patients with cancer.

HAP2 forms a capping structure, which binds very tightly to the distal end of flagellar filaments and still allows insertion of flagellin subunits below the cap by an unknown mechanism. Terminal regions of HAP2 from Salmonella typhimurium were found to be quickly degraded by various proteases, indicating that HAP2 also possesses disordered terminal regions like other axial proteins of bacterial flagellum. Removal of these portions by trypsin results in a fragment of 40 kDa (HP40), which lacks 42 NH2-terminal and 51 COOH-terminal residues. HAP2 in solution readily associates into a decameric structure without any significant population of intermediate oligomeric forms. The HP40 fragments, however, do not form decamers, while they can assemble into pentamers, as revealed by chemical cross-linking and analytical ultracentrifugation. Decameric HAP2 also dissociates into pentamers and smaller oligomers upon a heat induced conformational transition around 36 degreesC. While the highly mobile terminal regions are immobilized in decameric HAP2 complexes, they are still largely disordered in the pentameric state. These results demonstrate that the intersubunit interactions within the pentamers are mainly through the HP40 portions, whereas the terminal regions are responsible for association of pentamers into decameric complexes. Several observations indicate that HAP2 performs its capping function as a pentamer. We suggest that binding of the pentameric HAP2 cap to the filament is mediated by the highly flexible terminal regions. Indeed, HP40 fragments are unable to cap the end of filaments, while removal of about 30 residues from both terminal regions of HAP2 results in a highly reduced capping ability. A model is presented to explain the molecular mechanism of capping, in which conformational entropy in the disordered terminal regions moderates the otherwise too tight HAP2-filament interactions to allow insertion of flagellin subunits below the cap.

A new geranylated xanthone derivative, fuscaxanthone I (1), along with nine xanthones (2-9 and 11), a biphenyl (10) and three biflavonoids (12-14) were isolated from the roots of Garcinia fusca Pierre. Compounds 8, 10 and 11-14 were reported from this plant species for the first time. Their structures were elucidated by spectroscopic analyses, including 1D- and 2D-NMR and MS. The isolated compounds were evaluated for antibacterial activity against Helicobacter pylori. Cowaxanthone (5) and fukugiside (14) exhibited stronger inhibitory activity against H. pylori DMST reference strain at MICs 4.6 and 10.8 μM, respectively, than that of the control metronidazole. Isojacareubin (8) displayed the most potent activity against H. pylori HP40 clinical isolate with MIC 23.9 μM, which was approximately two times greater than that of the standard drug amoxicillin.

Seborrheic keratosis (SK) is a common benign epidermal tumor with predominance in adult patients. Whereas common SKs are more frequent in Caucasians, dermatosis papulosa nigra is more prevalent in patients with a Fitzpatrick skin type of at least 3. There seems to be a link between extrinsic skin aging and the occurrence of SK. Mutations of fibroblast growth factor receptor 3 and other signaling molecules are a frequent finding in SK lesions. However, this does not translate into any malignant potential. Viral infections are particularly common in genital lesions, although their pathogenetic relevance for SK is questionable. Different histologic and clinical subtypes have been identified. The great variability of SKs raises some difficulties in diagnosis. Dermoscopy is the preferred non-invasive diagnostic method, in particular to differentiate pigmented SKs from other pigment tumors, including cutaneous melanoma. Eruptive SKs can be a paraneoplastic condition known as the Leser-Trélat sign. New targeted cancer treatments can cause a pseudo-Leser-Trélat sign. The treatment in practice is mainly minor surgery, including cryosurgery, shave excisions, and laser-assisted removal. The medical approaches have only limited effects. Recently, two formulations for topical therapy have been evaluated: a product with 40% hydrogen peroxide (HP40) and an aqueous nitric-zinc complex. Based on clinical trials, HP40 seems to be a promising alternative to surgery, in particular for facial lesions.

BACKGROUND

Approved topical treatments for seborrheic keratoses (SKs) are an unmet need.

OBJECTIVE

To evaluate the safety and efficacy of 40% hydrogen peroxide topical solution (HP40) versus vehicle for the treatment of SKs (A-101-SEBK).

METHODS

A total of 937 patients with 4 SKs each (≥1 lesion each on the face and on the trunk and/or an extremity) were randomized 1:1 to HP40 or vehicle. At each visit, SKs were graded using the Physician's Lesion Assessment (PLA) scale (0, clear; 1, nearly clear; 2, ≤1 mm thick; and 3, >1 mm thick). After 1 treatment, SKs with a PLA score higher than 0 were re-treated 3 weeks later.

RESULTS

At day 106, significantly more patients treated with HP40 than with vehicle achieved a PLA score of 0 on all 4 SKs (study 1, 4% vs 0%; study 2, 8% vs 0% [both P < .01]) and on 3 of 4 SKs (study 1, 13% vs 0%; study 2, 23% vs 0% [both P < .0001]). A higher mean per-patient percentage of SKs were clear (study 1, 25% vs 2%; study 2, 34% vs 1%) and clear or nearly clear (study 1, 47% vs 10%; study 2, 54% vs 5%) with HP40 than with vehicle. Local skin reactions were largely mild and resolved by day 106.

CONCLUSIONS

The optimal number of treatment sessions was not evaluated.

CONCLUSIONS

Application of HP40 was well tolerated and effective in the removal of SKs.

OBJECTIVE

Seborrheic keratoses (SKs) may present in any non-glabrous skin, but data are limited on the response to treatment as based on the SK location. We aimed to understand the relationship between SK location and clearance with up to 2 treatments of 40% (w/w) hydrogen peroxide topical solution (HP40).

METHODS

We conducted a sub-analysis of data pooled from two randomized, double-blind, vehicle (VEH)-controlled clinical trials, including 937 patients, each with 4 target SKs (N=3,748 SKs), with at least 1 on the face and 1 on the trunk or extremities. Treatment response was defined as 0 or 1 on a 4-point Physician's Lesion Assessment (PLA) scale (0=clear; 1=near-clear) after up to 2 applications, 3 weeks apart, and was assessed by SK location (face, trunk, and extremity). Local skin reactions were stratified by anatomic location and categorized based on immediate and delayed post-treatment reactions. Sensitivity analysis was conducted using the mean-per-patient (MPP) percent of SKs that are clear or near-clear at day 106.

RESULTS

Treatment response was greater with HP40 versus VEH regardless of anatomic location of the SK. Clear or near-clear SKs with HP40 was observed in 65% of facial SKs (vs 10% VEH), 46% of truncal SKs (vs 5% VEH), and 38% of extremity SKs (vs 9% VEH). Facial SKs were more likely to be clear or near clear after a single treatment (43%), versus SKs on the trunk (31%) or extremities (14%). Most common immediate reactions with HP40 were erythema, stinging, and edema, which resolved to none or mostly mild within a week. Delayed reactions such as dyspigmentation and scarring occurred at low rates and were least reported for the facial SKs.

CONCLUSIONS

SK clearance with HP40 was highest among SKs on the face and lowest among SKs on the extremities. Dyspigmentation rates were lowest among SKs treated on the face. Anatomic location of SK was a predictor of both treatment response and risk of dyspigmentation with HP40 application. ClinicalTrials.gov listings: NCT02667236 and NCT02667275 J Drugs Dermatol. 2018;17(10):1092-1098.

Human interleukin-12(hIL-12) is a herterodimer cytokine, which consists of two disulfide-linked subunits, p40 and p35. This paper reports the expression of hIL-12 using the Baculovirus Expression System in insect cells. First, we constructed two expression vectors pVL1392-hp40 and pVL1393-hp35, and then they were used to co-transfect the insect cells (Sf9) separately with linearized polyhedrosis virus genomic DNA. Two kinds of recombinant viruses AcNPV-hp40 and AcNPV-hp35 were visually screened out. Biological activity of the recombinant hIL-12 (rhIL-12) was detected in the conditioned medium using proliferation assay of PHA-activated human lymphocytes and the expression level was about 1.5 approximately 2 microg/10(6) cells. The results of real-time Biomolecular Interaction Analysis (BIA) and Northern blot demonstrated that the subunits of rhIL-12, hp35, and hp40 were expressed successfully in the insect cells. The apparent molecular weights of rhIL-12 and hp40 homodimer were 76 kDa and 92 kDa under non-reducing conditions of Western blot, respectively. The recombinant hp40 can significantly inhibit the biological activity of hIL-12.

Seborrheic keratosis (SK) is an extremely common benign cutaneous lesion that often appears on individuals older than the age of 50. SK lesions can appear in numerous clinical variations, but typically, SKs present with a well-demarcated, “stuck-on” appearance having a waxy or keratotic surface. Even though SK lesions are usually benign, many individuals elect for SK treatment or removal due to cosmetic concerns. One study regarding SK appearance reported that 61% of women tried to conceal the appearance of the SKs by using certain hairstyles, make-up, and clothes. Conventional treatment for SK lesions ranges from cryosurgery to curettage. Although these removal methods are effective for SK treatment, each method contains a potential for side effects such as pigmentation changes, scarring, and discomfort before and after the procedure. Due to many of these concerns, patients prefer topical treatments for SKs, which has led to a new emerging topical containing hydrogen peroxide topical solution 40% (HP40; Eskata™). Here, we report the methods and results from an HP40 treatment for two individuals, each over 70 years old. Application of the topical solution was performed on each target SK, up to four times, with a 20-second application and 1 minute between applications. The two participants reported moderate success of HP40 in the removal of their SKs with minimal adverse effects.J Drugs Dermatol. 2019;18(7 Suppl):s178-182.

Objective: Hydrogen peroxide 40% (HP40) was approved by the US Food and Drug Administration for topical treatment of seborrheic keratosis (SK) in December 2017. This article will review phase II and III clinical trials to assess the drug's efficacy, safety, and clinical application. Data Sources: A systematic literature review was performed using the terms "Eskata AND seborrheic keratosis," and "hydrogen peroxide AND seborrheic keratosis" in the OVID MEDLINE, PubMed, Cochrane Library, EMBASE, and Web of Science databases. ClinicalTrials.gov was searched to identify ongoing or nonpublished studies. Study Selection and Data Abstraction: Articles written in English between January 2000 and mid-June 2020 discussing phase II and phase III clinical trials were evaluated. Data Synthesis: In 2 phase III clinical trials, 4% and 8% of patients treated with HP40 had a Physician Lesion Assessment score of zero for all 4 SKs, respectively, compared with 0% in both vehicle groups at the primary end point of day 106 (P < 0.01; P < 0.0001). Relevance to Patient Care and Clinical Practice: HP40, although less effective, has a better safety profile than other treatment options. It should be especially considered for treatment of facial SKs, where it is most efficacious and where other treatment modalities, such as cryotherapy, are more challenging. Conclusions: HP40 is a new, safe alternative treatment for SKs, although it is expensive and only modestly effective, both of which somewhat limit its overall utility. HP40 is a promising topical alternative, particularly for cosmetically sensitive locations, such as the face.

Keywords: adverse drug reactions; aging; clinical trials; cost benefit; dermatology; drug safety.

HP40 (Eskata™) is a stabilized, topical solution of 40% hydrogen peroxide (H2O2) packaged in an applicator pen that isUS FDA-approved to treat seborrheic keratoses (SKs). By harnessing the oxidative capabilities of H2O2 , 1-2 treatments with HP40 produced a higher rate of clearance of four SKs per patient compared to vehicle in two phase 3 trials. The clearance rate was higher for the face than the trunk and extremities. Similarly, the risks of pigmentary changes and scarring from HP40 were lower for the face than other locations. Further, based on an ex vivo study, HP40 may be less cytotoxic to melanocytes than cryotherapy, but clinical trials comparing these therapies are needed. Limitations of HP40 are its low efficacy and requirement of multiple treatments, which can result in elevated costs. The application can also be time-consuming, though extenders or even staff members can apply it. Therefore, HP40 may be better reserved for the treatment of facial SKs.

Despite reassurances about the benign nature of seborrheic keratoses (SKs), patients often request treatment due to cosmetic concerns or for symptomatic relief when SKs become irritated or pruritic. Treatment options include cryotherapy, surgical techniques, and topical therapies. In this study, we present two patients with SKs located on their face and neck who received in-office treatment with 40% Hydrogen Peroxide Topical Solution (Eskata™, HP40), a new FDA-approved topical therapy that has demonstrated efficacy in phase 3 trials. Compared to non-topical, more invasive techniques, HP40 may lead to less pigmentary changes, and may be more efficacious for SKs on the face and neck. Both patients received two treatment courses of HP40, which resulted in positive therapeutic outcomes, including the absence of scarring and pigmentary changes. In addition to the case presentations, we will discuss considerations for appropriate administration of HP40 to maximize clinical outcomes.J Drugs Dermatol. 2019;18(7 Suppl):s173-177.

Objectives: Assess participants&rsquo; satisfaction following treatment with a proprietary hydrogen peroxide topical solution 40%, w/w (HP40) for raised seborrheic keratoses (SKs).

Methods: In this Phase 4, open-label study, eligible participants aged 30&ndash;75 years had clinically typical raised SKs including 2 target SKs (Physician&rsquo;s Lesion Assessment&trade; [PLA] grade of &ge;2 [0 = clear; 1 = near clear; 2 = thin (&le;1 mm); 3 = thick (&gt;1 mm)]; 5&ndash;15 mm diameter) on the face and 1 target SK on the neck or d&eacute;colletage. SKs received HP40 treatment on day 1. All SKs with PLA grade &ge;1 were retreated on days 15 and 29. Endpoints included patients&rsquo; satisfaction with their skin&rsquo;s appearance at day 113, relationships between patients&rsquo; satisfaction and lesion PLA grade (evaluated by chi-square test), and patients&rsquo; satisfaction with their treatment experience.

Results: Forty-one patients (mean [range] age, 62.4 [46&ndash;73] years) completed the study. 95% of patients were at least moderately satisfied with their skin&rsquo;s appearance and 90.2% of target lesions were clear. A statistically significant association was observed between the number of target lesions achieving clearance and patients&rsquo; satisfaction with skin appearance level (&chi;2=22.03; P=0.001). 93% of patients were at least moderately satisfied with their HP40 treatment experience. Eight patients experienced treatment-emergent adverse events (TEAEs), most of which were mild or moderate; 4 experienced TEAEs considered treatment-related.

Conclusions: Most patients with SKs on the face, neck, and d&eacute;colletage were satisfied or very satisfied with both their skin&rsquo;s appearance and their treatment experience following HP40 treatment. These results support the use of HP40 for raised SKs. J Drugs Dermatol. 2020;19(12): doi:10.36849/JDD.2020.4974.

Objectives: The aims of this in vitro study was to investigate the effects of bleaching agents commonly used in micromorphology of the enamel surface and to assess the effect of concentration and of adding fluoride in the bleaching agents.

Methods: Sixty freshly extracted intact teeth were stored in distilled water. One half of each tooth was served as control, the other part was treated with bleaching agent. Samples were randomly divided into six groups of ten, according to the bleaching agents: G1- at-home-CP10; G2- at-home-CP16; G3- at-home-CP22; G4- in-office-CP35; G5- in-office-HP40 with fluoride; G6- in-office-HP40 without fluoride. Enamel specimens for each group were then submitted to a quantitative scanning electron microscopy. Number of pores and their diameter were measured to assess porosity of enamel surface.

Results: SEM analysis revealed enamel surface porosity after bleaching. Significant increase in number and major diameter of pores in bleached samples (p<0.001) were observed. The comparison between samples treated with 10% PC and samples treated with 22% PC showed significant increase in number of pores (p=0.006) and major diameter (p=0.001) from samples treated with 22% PC. Statistical analyses showed significant increase in the number of pores (p=0.006) from samples treated with 40% HP without fluoride compared to samples treated with 40% HP containing fluoride.

Conclusions: Bleaching products with low concentration cause less porosity at surface of the enamel compared to concentrated products. Adding fluoride in the bleaching agent appears to reduce porosity of enamel surface.

Keywords: Enamel; in vitro study; scanning electron microscopy; tooth morphology; vital bleaching.