Ultraviolet radiation stimulates the release of arachidonic acid from mammalian cells in culture

Abstract— C3H 10T½ cells in culture were prelabelled with [³H]arachidonic acid and exposed to UVB radiation. Almost immediately after irradiation cells released labelled arachidonate metabolites into media in a dose dependent manner. This release was inhibited by removing calcium ions from the system and by the addition of dexamethasone and parabromophenacyl bromide to the system. This suggests that the UVB stimulated release of arachidonic acid from membrane phospholipids is, in part, mediated by a phospholipase A₂ enzyme system. Thin layer chromatographic examination of media extracts revealed a dose dependent UVB stimulation of prostaglandin production by cultured cells.     

Ultraviolet B radiation induces upregulation of calcitonin gene-related peptide levels in human Finn chamber skin samples

One of the neuropeptides that plays a role in UVB-induced immunomodulation is calcitonin gene-related peptide (CGRP), as demonstrated in several animal studies. CGRP can be detected in human skin, but effects of UVB exposure on CGRP levels in human skin are not known. We determined CGRP levels in human Finn chamber skin samples of 15 UVB-irradiated and 10 control volunteers. Filter samples were collected prior to and immediately after a UVB exposure protocol (5 consecutive days, with one personally determined minimal erythema dose (MED(jp)) per day). CGRP levels in filter samples were determined using a commercially available radioimmunoassay kit. CGRP could be detected in the filter samples and volunteers showed statistically significantly increased levels after UVB exposure. In addition, the CGRP levels of UVB-exposed volunteers were positively correlated with the dose of UVB in J/m(2) that they received on 5 consecutive days. In other words, higher UVB doses resulted in higher CGRP levels. In summary, CGRP, a mediator in UVB-induced immunomodulation, could be detected in human Finn chamber skin samples, and was significantly increased after UVB exposure. The CGRP level appeared to depend on the amount of UVB the volunteers received.     

Acute Ultraviolet Radiation Perturbs Epithelialization but not the Biomechanical Strength of Full‐thickness Cutaneous Wounds

We hypothesized that priming of the skin with ultraviolet radiation (UVR) before being injured would enhance wound healing. Four groups, each comprising 20 immunocompetent hairless mice, were exposed to simulated solar irradiation in escalating UVR doses; 0 standard erythema dose (SED) = control, 1 SED, 3 SED and 5 SED. Twenty‐four hours after UV irradiation, inflammation was quantified by skin reflectance (erythema) and myeloperoxidase (MPO) tissue levels, and two 6 mm full‐thickness excisional wounds and one 3 cm incisional wound were inflicted. Epidermal hyperplasia was assessed by quantitative histology. Five days after wounding, wound coverage by neoepithelium and wound width of the excisional wounds was quantified in hematoxylin–eosin sections, and breaking strength was measured in strips from incisional wounds. Erythema (P < 0.001), MPO levels (P < 0.0005) and epidermal cell layers (P < 0.001) increased dose‐dependently by UV exposure of dorsal skin. In the excisional wounds, epithelial coverage decreased (P = 0.024) by increasing the UVR dose, whereas there was no significant difference (P = 0.765) in wound MPO levels. Neither wound width (P = 0.850) nor breaking strength (P = 0.320) differed among the groups. Solar‐simulated UVR 24 h before wounding impaired epithelialization but was not detrimental for surgical incisional wound healing.     

Ultraviolet A radiation induces rapid apoptosis of human leukemia cells by Fas ligand-independent activation of the Fas death pathways

Endogenous cellular chromophores absorb ultraviolet A radiation (UVA, 290-320 nm), the major UV component of terrestrial solar radiation, leading to the formation of reactive oxidizing species that initiate apoptosis, gene expression and mutagenesis. UVA-induced apoptosis of T helper cells is believed to underlie the UVA phototherapy for atopic dermatitis and other T cell-mediated inflammatory skin diseases. We have evaluated the involvement of the Fas-Fas ligand (FasL) pathway in rapid UVA-induced apoptosis in human leukemia HL-60 cells. UVA-induced apoptosis was not inhibited by pretreatment with a neutralizing anti-Fas antibody, although the same UVA treatment initiated cleavage of caspase-8 and subsequent processing of Bid and caspase-3-like proteases. Inhibition of caspase-8 by Lle-Glu (OMe)-Thr-Asp(OMe)-fluoromethyl ketone completely blocked caspase-3 cleavage and apoptosis in UVA-treated cells, suggesting that apoptosis was initiated by the Fas pathway. This inference was supported by demonstrating that immunoprecipitates obtained from UVA-treated cells using anti-Fas antibody contained caspase-8 and Fas-associating protein with death domain (FADD). In addition, Fas clustering in response to UVA treatment was observed by immunofluorescence microscopy. These data support a mechanism for rapid, UVA-induced apoptosis in HL-60 cells involving initial formation of the Fas-FADD-caspase-8 death complex in an FasL-independent manner.     

Ultraviolet radiation as a limiting factor in leaf expansion and development

Reductions in leaf growth are a commonly observed response to ultraviolet radiation, but the underlying mechanisms remain poorly defined. This study examined the response of leaves exposed to a UV environment across a range of organizational scales, including leaf expansion rate, epidermal cell size and number, biomechanical properties, leaf–water relations and activity of cell-wall peroxidases. Two experimental approaches were used; Lettuce ( Lactuca sativa L.) plants were propagated under (a) supplementary UV-B (9 kJ m⁻² day⁻¹) in controlled environment (CE) conditions, and (b) field conditions, where plants were placed under three horticultural films with differing UV transmissions. In both experiments, UV-B caused the greatest reductions in leaf expansion and final leaf size, with some reductions attributable to UV-A wavelengths. In supplementary UV-B conditions, adaxial cell size was reduced, while in field plants, both cell size and cell number were lower in an increased UV environment, as was the case with abaxial cells in CE plants. Although leaf turgor and leaf extensibility were not affected by UV wavelengths, breaking strain of leaf tissue was decreased under supplementary UV-B. Cell-wall peroxidase activity was increased in both supplementary UV conditions and in the field, where only a zero UV environment showed no upregulation of cell-wall peroxidase.     

Singlet oxygen, but not oxidizing radicals, induces apoptosis in HL-60 cells

Oxidizing species (OS), produced by photosensitization or derived from cytotoxic agents, activate apoptotic pathways. We investigated whether two different OS, formed at the same subcellular sites, have equivalent ability to initiate apoptosis in HL-60 cells. Our previous work showed that absorption of visible light by rose bengal (RB) produces singlet oxygen exclusively, whereas absorption of ultraviolet A produces RB-derived radicals in addition to singlet oxygen. Singlet oxygen, but not the RB-derived radicals, induced nuclear condensation and DNA fragmentation into nucleosome-size fragments in a dose dependent manner. In contrast, the RB-derived radicals caused greater lipid oxidation than singlet oxygen. These results indicate that different OS, produced at the same subcellular sites, do not have the same ability to induce apoptosis and that the ability of an OS to initiate lipid oxidation does not necessarily correlate with its ability to induce apoptosis.     

Antimony by ICP-MS as a marker for sentinel lymph nodes in melanoma patients

A sensitive, accurate and specific method for the analysis of antimony by ICP-MS is presented as a marker of the sentinel lymph node in melanoma patients.     

Ultraviolet radiation exposure of children and adolescents in Durban,South Africa

The solar ultraviolet radiation (UVR) exposure of 30 children and adolescents in three age groups (4-6 years, 7-9 years and 13-14 years) was measured for 1 week in late summer (February-March) in Durban, South Africa, using UVR-sensitive polysulfone film badges (PSFB) attached to the lapel region of the body. The mean and median values for all ages over the study period were 2.0 and 1.2 standard erythemal dose (SED) units, respectively, where 1 SED = 100 J x m(-2). Individual PSFB doses were analyzed as a function of age, gender and behavior. No significant statistical differences were found between different age groups; however, there was a statistical difference between males and females, with males generally receiving higher PSFB doses. Subjects completed UVR exposure journals documenting their time outdoors, shade versus sun conditions, nature of their activities, clothing worn and their use of sunscreen for each day of the study. Activity patterns were noted as the most important factor influencing individual UVR dose. Ambient erythemal UVR was measured by a Yankee Environmental Systems UVB pyranometer, and a relationship between ambient UVR and individual UVR dose was derived. On average, subjects received a dose of 4.6% of the total daily erythemal UVR. Based on this factor, the potential dose of an individual over a full annual cycle was estimated. Accordingly, there were 139 days during the year when an individual with skin type I (light skin) would be likely to experience minimal erythema and 97 and 32 days for individuals with skin types II and III, respectively.     

Decrease in langerhans cells and increase in lymph node dendritic cells following chronic exposure of mice to suberythemal doses of solar simulated radiation

Exposure of certain strains of mice to ultraviolet radiation (UVR) causes suppression of some innate and adaptive immune responses. One such consequence of acute UVB exposure is a reduction in the number of Langerhans cells (LC) in the epidermis and an increase in dendritic cells (DC) in lymph nodes draining the irradiated skin sites. Exposure to chronic UVB irradiation also has effects on the immune system, but it is unknown what effects are caused by repeated doses of solar simulated radiation (SSR). Consequently, the main aims of the present study were to determine whether repeated exposure to low doses of SSR would lead to similar changes in these cell populations and whether chronic doses of SSR activate a protective photoadaptation mechanism. Groups of C3H/HeN mice were irradiated daily with 3.7 J/cm(2) SSR from Cleo Natural lamps for 2, 10, 20, 30 or 60 days. Further groups of mice received an additional dose of 7.4 J/cm(2) SSR on days 2, 10, 30 or 60 to test for photoadaptation. The numbers of LC in the epidermis and DC in the lymph nodes draining irradiated skin sites were counted 24 h after the final irradiation. With the exception of mice irradiated for only 2 days, LC were significantly reduced throughout the chronic irradiation protocol, and no recovery occurred. DC numbers were significantly increased in the draining lymph nodes of mice irradiated for 20 days and 60 days.     

A self-assembled framework that interpenetrates in crystal but does not interpenetrate in solution

A porous supramolecular framework has been for the first time revealed to undergo interpenetration in crystal and noninterpenetration in solution. A new supramolecular organic framework Bu-SOF has been constructed from the co-assembly of a tetracationic tetrahedral monomer and cucurbit[8]uril(CB[8]) in water through the encapsulation of two anti-parallel n-butyl chains by CB[8]. X-ray diffraction analysis reveals that Bu-SOF forms 3-fold interpenetrated networks in crystals grown by evaporation of its solution in water.1 H NMR, dynamic light scattering and isothermal titration calorimetric experiments confirm that Bu-SOF is also formed in water. Solid samples, prepared by lyophilizing the aqueous solution of Bu-SOF, can adsorb nanoscaled organic dyes, supporting the porosity of the framework and thus non-interpenetration in solution. The avoidance of interpenetration of Bu-SOF in solution has been attributed to the filling of water inside the porous framework as well as the electrostatic repulsion of the appended bipyridinium units of the tetrahedral building block.     

A compartment of effector helper and suppressor T cells in the normal mouse thymus

We have recently described an autonomously activated set of T cells in the spleens of normal and ⪡antigen-free⪢ mice which display effector T helper (T_H) or T suppressor (T_S) activities; we describe here an intrathymic effector T-cell compartment which directly helps or suppresses B-cell responses and appears to be distinct from the peripheral set of effector cells. Splenic effector T cells do not represent recent thymic migrants (because adult thymectomized mice have unaltered levels of effector T_H and T_S cells in the spleen), nor do intrathymic effector T cells represent circulating peripheral T cells (since thymic effector T cells are B2A2⁺, while splenic effector T cells are B2A2⁻). Furthermore, effector T_H cells within the two compartments exert differential effector activities: splenic effector T_H cells induce B cells to both proliferation and maturation, while thymic effector T_H cells are defective in activating B-cell maturation. The present findings extend our studies on ⪡natural⪢ lymphocyte activities in the normal immune system, revealing the existence of two apparently distinct effector T-cell compartments. The potential significance of the intrathymic set of effector cells in repertoire selection is considered.     

Chronic mild stress decreases survival, but not proliferation, of new-born cells in adult rat hippocampus

New-born cells continue to proliferate and survive to become mature granule cells in adult rat hippocampus. Although this process, known as neurogenesis, is inhibited by acute stress, it is not clear whether chronic stress affects neurogenesis. To determine whether chronic mild stress (CMS) influences neurogenesis in the adult rat hippocampus, male Sprague-Dawley rats were exposed to CMS and administered bromodeoxyuridine (BrdU) before or after CMS to observe the survival/differentiation or proliferation of new-born cells, respectively. In addition, we measured brain-derived neurotrophic factor (BDNF) mRNA in the granule cell layer (GCL) of the hippocampus, because BDNF is known to play an important role in the survival of new-born cells. CMS significantly decreased the survival of new-born cells in the GCL, but did not influence the proliferation or differentiation of new-born cells. CMS did not affect the proliferation and survival of new-born cells in the hilus. In addition, CMS did not change BDNF mRNA levels in the GCL. These results demonstrate that CMS reduces the survival of new-born cells but not of their proliferation, suggesting that repeated mild stress could influence a part of neurogenesis, but not the whole part of neurogenesis. These results raise the possibility that the survival of new-born cells may be suppressed in the presence of normal BDNF mRNA levels in GCL.     

Ultraviolet radiation attenuates thrombospondin 1 expression via PI3K-Akt activation in human keratinocytes

Thrombospondin 1 (TSP1) is an extracellular glycoprotein and a recognized inhibitor of angiogenesis. Recent studies have demonstrated that UV radiation induces an angiogenic switch, by which it alters the balance between pro- and anti-angiogenic factors in the skin. Here we describe the effects of acute UV exposure on TSP1 expression in human skin epidermis, primary keratinocytes and the epidermal cell line HaCaT. We found that protein and mRNA expressions of TSP1 are significantly reduced in human skin in vivo and in keratinocytes in vitro by a single UV exposure. In human skin and keratinocytes, UV exposure induced the phosphorylation of Akt, a downstream target of the PI3K pathways. Specific inhibitors of PI3K, wortmannin and LY294002, completely blocked Akt activation and UV-induced TSP1 downregulation in keratinocytes. We showed that a specific Akt phosphorylation inhibitor and small interfering RNA-mediated Akt depletion were also blocked by UV-induced TSP1 downregulation in keratinocytes. In conclusion, our findings demonstrate that acute UV exposure downregulates TSP1 expression via PI3K-Akt activation in human keratinocytes. These novel findings may help us understand the regulatory mechanisms of UV-induced skin angiogenesis.     

Fumonisin B1 does not prevent apoptosis in A431 human epidermoid carcinoma cells after photosensitization with a silicon phthalocyanine

Photodynamic therapy with the phthalocyanine photosensitizer Pc 4 (Pc 4-PDT), an apoptosis inducer, is associated with accumulation of ceramide in various cell lines. The role of ceramide in Pc 4-PDT-induced apoptosis was investigated in A431 cells. Caspase-3 (casp-3) was activated and TUNEL positive cells began to appear 30 and 60 min post-Pc 4-PDT, respectively. A rapid increase (10 min) in cellular ceramide levels was observed after Pc 4-PDT. Induced ceramide accumulation was maintained over 60 min, Acid sphingomyelinase, a ceramide-generating enzyme, was inhibited after photosensitization with Pc 4, suggesting that the enzyme was not required for stimulated ceramide accumulation. Co-treatment of A431 cells with fumonisin B1, a ceramide synthase inhibitor, and Pc 4-PDT led to a decrease in ceramide levels without any effect on induced casp-3 activity or apoptosis. In the presence of zVAD, a pan-caspase inhibitor, apoptosis was abolished, while ceramide levels remained elevated after Pc 4-PDT. Exposure of A431 cells to exogenous C6-ceramide for 22 h, led to induction of apoptosis, and the process was abrogated by zVAD. In conclusion, C6-ceramide-, like Pc 4-PDT-induced apoptosis, is zVAD-sensitive. Furthermore, Pc 4 photosensitization can lead to apoptosis without FB-sensitive elevation in ceramide levels upstream of caspases.     

Photodynamic therapy resistant human colon carcinoma HT29 cells show cross-resistance to UVA but not UVC light

The isolation of photodynamic therapy (PDT)-resistant HT29 human colon adenocarcinoma cells has been reported previously. These PDT-resistant variants show increased expression of the Hsp27 and BNip3 proteins and a decreased expression of mutant p53 protein compared with parental HT29 cells. Because mutant p53 and increased expression of Hsp27 have been associated with resistance to various chemotherapeutic agents, whereas BNip3 is a potent inducer of apoptosis, we were interested in determining whether these PDT-resistant cells were cross-resistant to other cytotoxic agents. In the present report, we examined the colony survival of the PDT-resistant HT29 variants and several other clonal variants of HT29 cells to ultraviolet light (UV) treatment. The HT29 PDT-resistant variants showed cross-resistance to long-wavelength UVA (320-400 nm) but not to short-wavelength UVC (200-280 nm) light. Cell sensitivity to UVA or UVC was then correlated with Hsp27, BNip3 and mutant p53 protein levels in the PDT-resistant variants as well as in several clonal variants of HT29 cells that express different levels of Hsp27, BNip3 and mutant p53. We show that increased expression of Hsp27 and BNip3 and decreased expression of mutant p53 correlated with increased resistance to UVA. In contrast, increased expression of Hsp27 and BNip3 correlated with increased sensitivity to UVC, whereas increased expression of mutant p53 showed no significant correlation with sensitivity to UVC. These results suggest that the PDT-resistant HT29 cell variants are differentially sensitized to UVA compared with UVC due, in part at least, through the altered expression levels of BNip3, Hsp27 and mutant p53.     

Red/NIR emission carbonized polymer dots based on citric acid-benzoylurea and their application in lymph nodes imaging

Identification of lymph nodes (LNs) is critical for studies of the structure, the role in disease development, and the efficacy of disease treatment. Carbonized polymer dots (CPDs) are expected to be potential LNs-targeted imaging agents due to their excellent properties with special structure, better photoluminescence (PL) and great biocompatibility. Herein, a red/near infrared (NIR) emission CPDs (RCPDs) with one and two-photon bioimaging based on citric acid (CA) and benzoylurea (BU) are prepared. Notably, the RCPDs are capable of targeting LNs for imaging. Lymphocyte homing has been demonstrated to be the cellular mechanism of RCPDs target LNs imaging. This work has developed a new nanomaterial for targeted imaging of LNs, while the biological applications of CPDs have been expanded and deepened.     

Histopathological Changes in Liver and Heart Tissue Associated with Experimental Ultraviolet Radiation A and B Exposure on Wistar Albino Rats

This study aims to evaluate the influences of ultraviolet radiation A and B ( UVA + B) exposure on the liver and heart organs of albino rats. Female Wistar Albino rats, whose hair of the dorsal skin was shaved, were exposed to a combined UVA + B radiation for 2 h/day, for 4 weeks in order to be compared with the control group. Histopathological findings in vital organs (liver and heart) were evaluated. Tissues were fixed in 10% buffered formalin (pH = 7.2) and embedded in paraffin. The histopathological findings were examined on the H&E stained sections with light microscopy. The results show that the liver and the heart were injured in the UVA + B group. Liver tissue in the UVA + B group showed minimal vacuolation, enlargement of hepatocytes and bile duct proliferation, and the heart tissue showed hibernomas; uniform large cells resembling brown fat with coarsely granular to multivacuolated cytoplasm that is eosinophilic or pale with a small central nucleus. The number of hibernoma cases was significantly higher in the UVA + B group compared with the control group (P = 0.021). The control group showed normal liver and heart histology with normal adipose tissue in the pericardium. As a result, UVA + B exposure has toxic effects, especially on the liver and the heart of Wistar albino rats. UV radiation may cause such adverse effects in humans. Therefore, protection against the harmful effects of UV radiation is of significant importance for skin and organs.     

Why tert-butyl alcohol associates in aqueous solution but trimethylamine-N-oxide does not

In dilute aqueous solution, tert-butyl alcohol (TBA) tends to aggregate but trimethylamine-N-oxide (TMAO) does not. Given that both molecules have very similar geometry with hydrophobic and hydrophilic groups, it is interesting to ask why they behave so differently in aqueous solution. To explore this question, we use molecular dynamics simulations to study two models representing TBA and TMAO that differ essentially only in their electrostatic properties. It is shown that this difference is sufficient to give the different solution behavior. Furthermore, the principal difference identified is that the hydrophilic group of TMAO (the oxygen) interacts on average much more strongly with water than the corresponding group (the hydroxyl) of TBA. A hydrogen-bond analysis shows that water-TBA and water-TMAO hydrogen bonds are similar in number, but that the hydrogen-bond energy is much more negative for water-TMAO. In all likelihood, this accounts for the different behavior in dilute aqueous solution.     

The sulphate radical is not involved in aqueous radiation oxidation processes

The H₂O₂/persulphate systems are of enormous environmental and commercial importance with the sulphate radical (SO₄⁻) being assumed as the oxidizing/bleaching species. We show that under normal conditions (air-saturated) no SO₄⁻ is produced and, most likely, a much longer-lived species, the adduct of O₂ and the persulphate radical, is formed.