The Endogenous Tryptophan‐derived Photoproduct 6‐formylindolo[3,2‐b]carbazole (FICZ) is a Nanomolar Photosensitizer that Can be Harnessed for the Photodynamic Elimination of Skin Cancer Cells in Vitro and in Vivo

UV‐chromophores contained in human skin may act as endogenous sensitizers of photooxidative stress and can be employed therapeutically for the photodynamic elimination of malignant cells. Here, we report that 6‐formylindolo[3,2‐b]carbazole (FICZ), a tryptophan‐derived photoproduct and endogenous aryl hydrocarbon receptor agonist, displays activity as a nanomolar sensitizer of photooxidative stress, causing the photodynamic elimination of human melanoma and nonmelanoma skin cancer cells in vitro and in vivo. FICZ is an efficient UVA/Visible photosensitizer having absorbance maximum at 390 nm (ε = 9180 L mol^?1 cm^?1), and fluorescence and singlet oxygen quantum yields of 0.15 and 0.5, respectively, in methanol. In a panel of cultured human squamous cell carcinoma and melanoma skin cancer cells (SCC‐25, HaCaT‐ras II‐4, A375, G361, LOX), photodynamic induction of cell death was elicited by the combined action of solar simulated UVA (6.6 J cm^?2) and FICZ (≥10 nm ), preceded by the induction of oxidative stress as substantiated by MitoSOX Red fluorescence microscopy, comet detection of Fpg‐sensitive oxidative genomic lesions and upregulated stress response gene expression (HMOX1, HSPA1A, HSPA6). In SKH1 “high‐risk” mouse skin, an experimental FICZ/UVA photodynamic treatment regimen blocked the progression of UV‐induced tumorigenesis suggesting feasibility of harnessing FICZ for the photooxidative elimination of malignant cells in vivo.     

Distinction of malignant melanoma and epidermis using IR micro-spectroscopy and statistical methods

Infrared spectroscopy is widely perceived as a future technology for cancer detection and grading. Malignant melanoma, an aggressive skin cancer, is accessible to non-invasive IR radiation based surface probes for its identification and grading. The present work examines the differences in the IR spectra of melanoma tissues and the surrounding epidermis in skin biopsies with the objective of identifying diagnostic parameters and suitable computational/statistical methods of analysis. Melanoma could be differentiated from the epidermis in biopsies of 55 patients, using parameters derived from absorbance bands originating from molecular vibrations of nucleic acids and/or their bases. Additionally, absorbances from tyrosine and phosphate that are abnormally elevated in malignant melanoma could be used as markers. Two-dimensional plots of these parameters in tandem with advanced statistical methods successfully demonstrate the potential of IR spectroscopy to distinguish between epidermal and melanoma regions with a high classification success. The work underlines the importance of developing data analysis methods in FTIR based diagnosis using melanoma as a model system.     

Skin Cancer Risks Avoided by the Montreal Protocol—Worldwide Modeling Integrating Coupled Climate‐Chemistry Models with a Risk Model for UV

The assessment model for ultraviolet radiation and risk “AMOUR” is applied to output from two chemistry‐climate models (CCMs). Results from the UK Chemistry and Aerosols CCM are used to quantify the worldwide skin cancer risk avoided by the Montreal Protocol and its amendments: by the year 2030, two million cases of skin cancer have been prevented yearly, which is 14% fewer skin cancer cases per year. In the “World Avoided,” excess skin cancer incidence will continue to grow dramatically after 2030. Results from the CCM E39C‐A are used to estimate skin cancer risk that had already been inevitably committed once ozone depletion was recognized: excess incidence will peak mid 21st century and then recover or even super‐recover at the end of the century. When compared with a “No Depletion” scenario, with ozone undepleted and cloud characteristics as in the 1960s throughout, excess incidence (extra yearly cases skin cancer per million people) of the “Full Compliance with Montreal Protocol” scenario is in the ranges: New Zealand: 100–150, Congo: ?10–0, Patagonia: 20–50, Western Europe: 30–40, China: 90–120, South‐West USA: 80–110, Mediterranean: 90–100 and North‐East Australia: 170–200. This is up to 4% of total local incidence in the Full Compliance scenario in the peak year.     

Skin Cancer Awareness Among 1 271 Black Africans in South Africa

Little is known about levels of awareness and perceptions of skin cancer among Africans living in Africa. This study assessed skin cancer awareness among 1271 deeply pigmented South Africans. Participants (n = 642 males vs n = 629 females) were aware of skin cancer (79%) with more females than males being aware of skin cancer (P = 0.02). Majority of all participants had never checked their skin for signs or symptoms of skin cancer (90%). Palms of hands and soles of feet were the least recognized anatomic sites for skin cancer development, despite these sites being the common sites for acral lentiginous melanoma in individuals with deeply pigmented skin. Results suggest a need for targeted skin cancer awareness among population groups with dark skin on identification, screening, and early detection, professional training for healthcare personnel and content on skin cancer in deeply pigmented skin in medical curricula.     

Beyond the BET Family: Targeting CBP/p300 with 4‐Acyl Pyrroles

Bromodomain and extra‐terminal domain (BET) inhibitors are widely used both as chemical tools to study the biological role of their targets in living organisms and as candidates for drug development against several cancer variants and human disorders. However, non‐BET bromodomains such as those in p300 and CBP are less studied. XDM‐CBP is a highly potent and selective inhibitor for the bromodomains of CBP and p300 derived from a pan‐selective BET BRD‐binding fragment. Along with X‐ray crystal‐structure analysis and thermodynamic profiling, XDM‐CBP was used in screenings of several cancer cell lines in vitro to study its inhibitory potential on cancer cell proliferation. XDM‐CBP is demonstrated to be a potent and selective CBP/p300 inhibitor that acts on specific cancer cell lines, in particular malignant melanoma, breast cancer, and leukemia.     

Induction of primary cutaneous melanomas in C3H mice by combined treatment with ultraviolet radiation, ethanol and aloe emodin

The role of ultraviolet (UV) radiation in the induction of nonmelanoma skin cancer is widely accepted, although its precise contribution to the development of primary cutaneous melanoma skin cancer requires further definition. We found that painting aloe emodin, a trihydroxyanthraquinone from Aloe barbadensis, in ethyl alcohol vehicle on the skin of mice in conjunction with exposure to UVB (280-320 nm) radiation results in the development of melanin-containing skin tumors. C3H/HeN mice were treated thrice weekly with aloe emodin in a 25% ethanol in water vehicle and exposed to 15 kJ/m2 UV radiation. Neither ethanol vehicle nor aloe emodin alone induced skin tumors in the absence of UV radiation. In two separate experiments, 20-30% of the mice treated with a combination of UV radiation and ethanol vehicle and 50-67% of the UV-irradiated animals given aloe emodin in ethanol vehicle developed primary cutaneous melanin-containing tumors. The diagnosis of melanoma was established using Fontana silver stain for melanin; these tumors were negative for vimentin and keratin. Melanin-containing melanosomes were observed by transmission electron microscopy in tumors diagnosed as melanomas. Although the mechanism of carcinogenesis in these mice is currently unknown, our findings have led to the development of the first facile murine model for the induction of primary melanoma. This model has the potential to clarify the role of UV radiation in the etiology of malignant melanoma.     

An Experimental Population Study of Nucleotide Excision Repair as a Risk Factor for UVB‐induced Melanoma

Nucleotide excision repair (NER) is the primary defense against the DNA damage implicit in skin cancer formation and is negatively affected by chronic exposure to UVB radiation. However, in situ and in vitro studies consistently yield equivocal results when addressing individual DNA repair capacity and melanoma susceptibility. The primary objective of this study was to determine if individual global NER capacity is a risk factor for melanoma formation in a prominent UVB‐inducible melanoma model, hybrid Xiphophorus fishes. After neonatal UVB irradiation, adult tumor‐bearing and tumor‐free fish were given a challenge UVB dose and (6–4) photoproduct repair was quantified in individual fish at 24 h using radioimmunoassay. Despite considerable inter‐individual variation in repair capacity, ranging from 13% to 91%, we found no difference in mean NER capacity between fish with and without melanomas, thus detaching global NER from melanomagenesis. Furthermore, despite epidemiological data indicating that sex and age are important risk factors underlying melanoma susceptibility, we found no difference in mean NER rates among the sexes or as a function of age. We conclude with a discussion of the apparent paradox of how inter‐individual variation in NER is not a risk factor given the clear evidence that DNA damage underlies melanoma susceptibility.     

123I-N-isopropyl-p-iodoamphetamine scintigraphy in patients with malignant melanoma

N-Isopropyl-p-[123I]iodoamphetamine (123I-IMP) has been widely used in patients with cerebral vascular damage and epilepsy. The potential of 123I-IMP accumulation in the melanin producing cells has been reported since early stage of its development. Some authors reported the accumulation of 123I-IMP in the malignant melanoma of human and animals. We evaluated the 123I-IMP scintigraphy in 9 patients with malignant melanoma. Patients were classified into two groups: A, 4 patients with 8 lesions; B, 5 post-operative patients without lesions. In group A, the 123I-IMP uptake was seen in 4 of 8 lesions. The smallest true positive lesion was located at skin and its size was 15 mm in diameter. Two of the visualized 4 lesions were amelanotic malignant melanomas. This fact suggests that uptake of 123I-IMP in malignant melanoma may be related to the processes of melanin synthesis. In group B, two abnormal deposits had been seen in the right thigh of a female patient. However no abnormality was seen in the following 67Ga scintigraphy, TCT, MRI, and repeated 123I-IMP scintigraphy. Therefore the abnormal deposits were considered to be the false positive lesions due to urinary contamination.     

The Monodelphis melanoma model: initial report on large ultraviolet A exposures of suckling young

The objective of this study was to determine whether exposure of early suckling young of the opossum Monodelphis domestica to ultraviolet A (UVA) radiation (320-400 nm) can lead to the development of melanocytic lesions similar to those induced after exposure to ultraviolet B (UVB) radiation (280-320 nm) to total doses as low as 380 J/m2. A total of 576 sucklings received nine exposures of 0.6, 2.6 or 15.5 kJ/m2 per dose (total doses approximately 6, 23 and 140 kJ/m2, respectively) from a Blak Ray lamp source with a narrow range emission at 365 nm. A further 280 sucklings were exposed in the same way to doses of 2.6 kJ/m2 per dose (total approximately 23 kJ/m2) broad-band UVA with visible wavelengths from a Dermalight lamp. Frequency of litter loss following all of the UVA-exposure protocols was similar to that within the same stocks in the colony at large. Only one of the 856 UVA-exposed individuals possessed a melanocytic lesion at the 5 month assessment point. No radiation-induced lesions of any type were evident on the skin of the other animals exposed as sucklings. The affected male was from a group of 70 individuals exposed to the highest total dose (140 kJ/m2) from the Blak Ray light source. The melanocytic hyperplasia was provisionally identified as a potential melanoma but it slowly regressed as the animal aged. We conclude that in the opossum suckling exposure system, the potency of UVA for melanoma induction is extremely low compared with that of UVB. Possible explanations, amenable to further investigations, are given for the low UVA sensitivity of the suckling model compared to the adult exposure model of Ley (Ley, R. D. [1997] Cancer Res. 57, 3682-3684).     

In Vivo Optical Properties of Melanocytic Skin Lesions: Common Nevi, Dysplastic Nevi and Malignant Melanoma

We present an in vivo study of the optical properties of common nevi, dysplastic nevi and malignant melanoma skin lesions in human subjects. Reflectance spectra were measured on 1379 skin lesions, in the visible and near-infrared spectral regions, using a spectral imaging system, in a clinical setting. Analysis of the data using a reflectance model revealed differences between the optical properties of melanin present in nevi and melanoma lesions. These differences, which are in agreement with our previous observations on average reflectance spectra, may be potentially useful for the noninvasive characterization of pigmented skin lesions and the early diagnosis of melanoma.     

In vivo SPECTROPHOTOMETRIC EVALUATION OF NEOPLASTIC AND NON-NEOPLASTIC SKIN PIGMENTED LESIONS. II: DISCRIMINANT ANALYSIS BETWEEN NEVUS AND MELANOMA

Reflectance spectrophotometry from 420 to 780 nm on 31 primary melanoma and 31 benign nevi has been performed by using an external integrating sphere coupled to a spectrophotometer. Measurements show that reflectance spectra of melanoma and nevi manifest dissimilar patterns. From these spectra four variables, whose physical and/or physiological meanings remain to be investigated, have been derived. All of them are significantly different when compared between melanoma and nevi. A discriminant function between the two groups of lesions has been determined by using a stepwise discriminant analysis, resulting in a test with a sensitivity of 90.3% and a specificity of 77.4%. This method of discrimination between melanoma and nevi seems to have a discriminating power almost equal to that of a clinical judgement from a specialized medical doctor, thus suggesting a new method for screening skin pigmented lesions.     

Distinct Role of Sesn2 in Response to UVB‐Induced DNA Damage and UVA‐Induced Oxidative Stress in Melanocytes

Ultraviolet (UV) radiation, including both UVB and UVA irradiation, is the major risk factor for causing skin cancer including melanoma. Recently, we have shown that Sesn2, a member of the evolutionarily conserved stress‐inducible protein family Sestrins (Sesn), is upregulated in human melanomas as compared to melanocytes in normal human skin, suggesting an oncogenic role of Sesn2. However, the role of Sesn2 in UVB and UVA response is unknown. Here, we demonstrated that both UVB and UVA induce Sesn2 upregulation in melanocytes and melanoma cells. UVB induces Sesn2 expression through the p53 and AKT3 pathways. Sesn2 negatively regulates UVB‐induced DNA damage repair. In comparison, UVA induces Sesn2 upregulation through mitochondria but not Nrf2. Sesn2 ablation increased UVA‐induced Nrf2 induction and inhibits UVA‐induced ROS production, indicating that Sesn2 acts as an upstream regulator of Nrf2. These findings suggest previously unrecognized mechanisms in melanocyte response to UVB and UVA irradiation and potentially in melanoma formation.     

超声造影和MRI在胃癌新辅助治疗后再分期的一致性

将2017年1月至2018年12月于西宁市第二人民医院行新辅助治疗的72例胃癌患者纳入研究,同时给予患者超声造影(contrast-enhanced ultrasonography,CEUS)检查及磁共振成像(magnetic resonance imaging, MRI)增强扫描,比较并分析二者对治疗后分期及肿瘤周围侵犯情况的检查结果,以探究CEUS在评估胃癌新辅助治疗后分期及侵犯情况方面与MRI的一致性。研究结果显示,MRI和CEUS评估胃癌T分期的诊断一致率为91.67%,具有较高一致性;CEUS和MRI评估病灶的左右径、纵轴直径和前后径具有较好的一致性,且两者无明显差异;CEUS与MRI评估其侵袭横结肠及其系膜、肝/脾、十二指肠/胰腺的诊断,具有较高的一致性。本研究证实,CEUS与MRI在评估胃癌T分期、病灶大小及周围组织侵犯情况时结果基本一致。     

A validated high‐performance thin‐layer chromatography method for the identification and simultaneous quantification of six markers from Platanus orientalis and their cytotoxic profiles against skin cancer cell lines

Betulinic acid ( 1 ), betulinic acid‐3‐acetate ( 2 ), 3‐acetylbetulinaldehyde ( 3 ), oleanolic acid‐3‐acetate ( 4 ), 3‐β‐hydroxy‐28,19‐β‐olenolide ( 5 ), and β‐sitosterol ( 6 ) were isolated from Platanus orientalis and a high‐performance thin‐layer chromatography method was developed for their simultaneous quantification. The markers were first derivatized on the chromatogram with ceric ammonium sulfate and then high‐performance thin‐layer chromatography densitometry was carried out. Chromatographic separation of these markers was carried out on silica gel 60 plates using a ternary solvent system n‐hexane/toluene/acetone (6:3.5:1 v/v/v) as a mobile phase. For marker 1 , a deuterium (D₂) lamp and wavelength of 420 nm was used. A tungsten (W) lamp was used for markers 2 and 3 at 550 nm and for 4 – 6 at 500 nm. The method was validated for accuracy, precision, LOD, and LOQ. All calibration curves showed a good linear relationship (r > 0.9919). The precision evaluated by an intra‐ and interday study showed RSDs < 2.51% and accuracy validation recovery between 95.54 and 99.33% with RSDs < 1.55%. The successful application of the validated method showed 1 as the most abundant component (4.63%) and 5 (0.017%) the least. The markers displayed a significant cytotoxic effect against human keratinocyte, mouse melanoma, and human skin epithelial carcinoma cancer cells by using a 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay.     

Hydrogel‐Forming and Dissolving Microneedles for Enhanced Delivery of Photosensitizers and Precursors

We present “one‐step application” dissolving and hydrogel‐forming microneedle arrays (MN) for enhanced delivery of photosensitizers/precursors. MN (280 μm) prepared from 20% w/w poly(methylvinylether/maelic acid) and cross‐linked with glycerol by esterification to form hydrogels upon skin insertion, or allowed to dissolve rapidly in skin, were combined with patches containing 19 mg cm^?2 of 5‐aminolevulinic acid (ALA) or meso‐tetra (N‐methyl‐4‐pyridyl) porphine tetra tosylate (TMP) for drug delivery. Both MN types were mechanically robust, with compression forces of 20.0 N only causing height reductions of 14%. Application forces as low as 8.0 N per array allowed >95% of the MN in each array type to penetrate excised porcine skin, with the MN penetrating to approximately 220 μm. MN significantly enhanced transdermal delivery of ALA and TMP in vitro, with the hydrogel‐forming system comparable with the dissolving system for ALA delivery (approximately 3000 nmol cm^?2 over 6 h), but superior for delivery of the much larger TMP molecule (approximately 14 nmol cm^?2 over 24 h, compared to 0.15 nmol cm^?2). As this technology clearly has potential in enhanced photodynamic therapy of neoplastic skin lesions, we are currently planning animal studies, to be followed by preliminary human evaluations. GMP manufacturing scale‐up is ongoing.     

Noninvasive Optical Imaging of UV‐Induced Squamous Cell Carcinoma in Murine Skin: Studies of Early Tumor Development and Vitamin D Enhancement of Protoporphyrin IX Production

Better noninvasive techniques are needed to monitor protoporphyrin IX (PpIX) levels before and during photodynamic therapy (PDT) of squamous cell carcinoma (SCC) of the skin. Our aim was to evaluate (1) multispectral fluorescent imaging of ultraviolet light (UV)‐induced cancer and precancer in a mouse model of SCC and (2) multispectral imaging and probe‐based fluorescence detection as a tool to study vitamin D (VD) effects on aminolevulinic acid (ALA)‐induced PpIX synthesis. Dorsal skin of hairless mice was imaged weekly during a 24‐week UV carcinogenesis protocol. Hot spots of PpIX fluorescence were detectable by multispectral imaging beginning at 14 weeks of UV exposure. Many hot spots disappeared after cessation of UV at week 20, but others persisted or became visible after week 20, and corresponded to tumors that eventually became visible by eye. In SCC‐bearing mice pretreated with topical VD before ALA application, our optical techniques confirmed that VD preconditioning induces a tumor‐selective increase in PpIX levels. Fluorescence‐based optical imaging of PpIX is a promising tool for detecting early SCC lesions of the skin. Pretreatment with VD can increase the ability to detect early tumors, providing a potential new way to improve efficacy of ALA‐PDT.     

Application of probabilistic neural network in the clinical diagnosis of cancers based on clinical chemistry data

As a recently developed and powerful classification tool, probabilistic neural network was used to distinguish cancer patients from healthy persons according to the levels of nucleosides in human urine. Two datasets (containing 32 and 50 patterns, respectively) were investigated and the total consistency rate obtained was 100% for dataset 1 and 94% for dataset 2. To evaluate the performance of probabilistic neural network, linear discriminant analysis and learning vector quantization network were also applied to the classification problem. The results showed that the predictive ability of the probabilistic neural network is stronger than the others in this study. Moreover, the recognition rate for dataset 2 can achieve to 100% if combining these three methods together, which indicated the promising potential of clinical diagnosis by combining different methods.     

Detection of non‐small cell lung cancer cells based on microfluidic polarization microscopic image analysis

In early diagnosis of lung cancer, a polarization microscopy is a powerful tool to obtain the optical information of biological tissues. In this paper, a new microfluidic polarization imaging and analysis method was proposed for the detection and classification of cancer‐associated fibroblasts and the two kinds of non‐small cell lung cancer cells, A549 and H322. A polarizing microscopy system was constructed based on a commercial microscope to obtain 3*3 Mueller matrix of cells. Based on the Muller matrix decomposition algorithm and analysis in spatial domain and frequency domain, appropriate classification parameters were selected for the characterization of different polarization characteristics of cells. Finally, the logistic regression models based on machine learning were applied to determine optimal feature parameters and classify cells. This method integrated the morphological information of the cells, and the polarization characteristics of the cells in different polarization states. It is for the first time that the polarization microscopic image analysis method has been applied to the detection and classification of non‐small cell lung cancer cells. The results show that the presented microfluidic polarization microscopic image analysis method could classify cells effectively. Compared with the Muller matrix measurement and calculation methods, the method proposed in this paper was greatly simplified in both the acquisition of polarized images and the analysis and processing of polarized images.     

Characteristics of Xeroderma Pigmentosum in Japan: Lessons From Two Clinical Surveys and Measures for Patient Care

Xeroderma pigmentosum (XP) is a rare autosomal recessive hereditary disease caused by deficiency in repair of DNA lesions generated by ultraviolet radiation and other compounds. Patients with XP display pigmentary change and numerous skin cancers in sun‐exposed sites, and some patients show exaggerated severe sunburns even upon minimum sun exposure as well as neurological symptoms. We conducted a nationwide survey for XP since 1980. In Japan, the frequency of the XP complementation group A is the highest, followed by the variant type; while in the Western countries, those of groups C or D are the highest. Regarding skin cancers in XP, basal cell carcinoma was the most frequent cancer that afflicted patients with XP, followed by squamous cell carcinoma, and malignant melanoma. The frequency of these skin cancers in patients with XP has decreased in these 20 years, and the age of onset of developing skin cancers is higher than those previously observed, owing to early diagnosis and education to patients and care takers on strict prevention from sunlight for patients with XP. On the other hand, the effective therapy for neurological XP has not been established yet, and this needs to be done urgently.