Quercetagetin inhibits macrophage-derived chemokine in HaCaT human keratinocytes via the regulation of signal transducer and activator of transcription 1, suppressor of cytokine signalling 1 and transforming growth factor-ß1.

BACKGROUND: Inflammatory chemokines, such as macrophage-derived chemokine (MDC/CCL22), are elevated in the serum and lesioned skin of patients with atopic dermatitis (AD), and are ligands for C-C chemokine receptor 4, which is predominantly expressed on T helper 2 lymphocytes, basophils and natural killer cells. We have previously reported that quercetagetin has an inhibitory activity on inflammatory chemokines, which is induced by interferon (IFN)-γ and tumour necrosis factor (TNF)-α, occurring via inhibition of the signal transducer and activator of transcription 1 (STAT1) signal. OBJECTIVES: To investigate the specific mechanisms of quercetagetin on the STAT1 signal. METHODS: We confirmed the inhibitory activity of quercetagetin on MDC and STAT1 in HaCaT keratinocytes. The interaction between STAT1 and IFN-γR1 was investigated using immunoprecipitation. The small interfering RNA approach was used to investigate the role of suppressor of cytokine signalling 1 (SOCS1) and transforming growth factor (TGF)-ß1 induced by quercetagetin. RESULTS: Quercetagetin inhibited the expression of MDC at both the protein and mRNA levels in IFN-γ- and TNF-α-stimulated HaCaT human keratinocytes. Moreover, quercetagetin inhibited the phosphorylation of STAT1 through upregulation of SOCS1. Increased expression of SOCS1 disrupted the binding of STAT1 to IFN-γR1. Furthermore, quercetagetin augmented the expression of TGF-ß1, which is known to modulate the immune response and inflammation. CONCLUSIONS: These results suggest that quercetagetin may be a potent inhibitor of the STAT1 signal, which could be a new molecular target for anti-inflammatory treatment, and may thus have therapeutic applications as an immune modulator in inflammatory diseases such as AD.

Oncologic outcomes of laparoscopic nephroureterectomy for pT3 upper urinary tract urothelial carcinoma.

AIM: We present the oncologic outcomes of laparoscopic nephroureterectomy management of pT3 upper urinary tract urothelial carcinoma. METHODS: Between October 2003 and January 2011, 50 patients with pT3 upper urinary tract urothelial carcinoma which had pathologically confirmed underwent laparoscopic nephroureterectomy at our institution. Demographic data, perioperative results, pathological findings and oncologic outcomes were reviewed and analyzed retrospectively. RESULTS: There were 36 patients (72%) of high grade lesion and 14 patients (28%) of low grade lesion. Lymphovascular invasion was observed in 16 patients (32%) and the surgical margin was positive in one patient. N stage was pN0 in 16 (32%), pN1 in 3 (6%), pN2 in 1 (2%) and pN3 in 1 (2%). The 5-year overall survival rate was 52.6% and the 5-year cancer-specific survival rate was 65.3%. Overall recurrence developed in 23 patients. There were 10 patients (20%) of urothelial recurrence which were all occurred in the bladder at the mean period of 13.6 months, and 7 patients of them were invasive bladder cancer. There were 16 patients (32%) of non-urothelial recurrence developed at the mean period of 9.69 months. On multivariate analyses lymphadenopathy and lymph node involvement of cancer (N+) were identified as independent predictive factors for the cancer-specific survival, and concomitant bladder tumor, grade and lymphovascular invasion were identified as independent predictive factors for the overall recurrence free survival. CONCLUSION: Laparoscopic nephroureterectomy in patients with high stage upper urinary tract urothelial carcinoma appear comparable to those of open surgery in the regard of oncologic outcomes.

Resistive switching characteristics of the Cr/ZnO/Cr structure.

Resistive random access memory (ReRAM) with conductor-dielectric-conductor structures has attracted extensive attention for next generation nonvolatile memory devices. The resistive switching effect has been observed in various materials, such as metal oxides and chalcogenide oxides. From our findings, we advocate the resistive switching characteristics of zinc oxide thin film, due to its simple composition and ease of manipulation. In this study, we investigated the current-voltage (I-V) characteristics of the Cr/ZnO/Cr capacitor structure. The Cr electrode and ZnO thin film were deposited by radio frequency magnetron sputtering at room temperature. The top electrode layers were patterned by 100 microm x 100 microm. The fabricated devices of the Cr/ZnO/Cr structures exhibited bipolar switching behavior. In addition, using the Cr-coated AFM tip replaced with the top electrode enabled us to map the local current image and measure the current flow at each point. This gave us more information to verify the resistive switching mechanism of ZnO thin film.

Factors affecting completion of laparoscopic myomectomy.

PURPOSES: This study aimed to elucidate the factors affecting completion of laparoscopic myomectomy without unintended surgery. MATERIALS AND METHODS: The medical records of 143 patients who underwent laparoscopic myomectomy desiring to retain their uterus were retrospectively reviewed. Unintended surgery was defined as the need for conversion to other surgical methods including laparotomy or laparoscopic hysterectomy at any time during the procedures. All variables associated with completion of laparoscopic myomectomy in the univariate analysis were selected at the threshold ofp < 0.25 and then tested in a multiple-logistic regression model. RESULTS: The rate of unintended surgery was 13.3%. Univariate analysis revealed that age, previous abdomino-pelvic surgery, current medical disease, transfusion, > five myomas, myoma size > 8.2 cm, posterior wall location of myoma, intramural type of myoma, and the presence of adenomyosis were statistically significant risk factors for unintended surgery. Multivariate logistic regression analysis demonstrated that completion of laparoscopic myomectomy was significantly influenced by a history of previous abdomino-pelvic surgery (odds ratio; 6.46, 95% CI, 0.03-0.41; p value 0.04). CONCLUSION: The risk of unintended surgery during laparoscopic myomectomy is associated with a history of previous abdomino-pelvic surgery.

The effects of cranio-cervical flexion on activation of swallowing-related muscles.

We tested the effects of cranio-cervical flexion (CCF) on activation of swallowing-related muscles while swallowing liquid in a sample of 45 healthy volunteers. Activation following CCF movement was examined across two positions (supine and sitting) and, three pressure levels and two different postures were examined in each condition, respectively. When CCF was applied, activation of swallowing-related muscles was significantly increased compared to the neutral neck position, and such findings were found across both the supine and sitting positions. Also in the supine position, when the pressure level of the stabilizer was escalated, there was a significant difference in the activity of the swallowing-related muscles compared to the baseline level. In conclusion, our results suggest that CCF may be a viable method to enhance the effectiveness of swallowing-related muscles by changing neck position. When CCF is applied, the stability of the deep flexor muscles must be secured first after which superficially located muscles may better assist swallowing with less effort.

Predictive value of obstructive voiding symptoms and objective bladder emptying tests for urinary retention.

The prevalence of voiding dysfunction was 47.4% by the question of "do you usually experience a feeling of incomplete bladder emptying" from Korean version of PFDI (pelvic floor distress inventory), 33.7% by the value of less than 10th centile of peak flow rate of uroflowmetry by Liverpool nomogram, 20.2% by the cutoff values of less than 12 ml per second of maximum flow rate and greater than 25 cmH20 of detrusor pressure at maximum flow rate in the pressure flow study and 11.6% by the elevated post-void residual urine volume more than 50 ml in 95 women who visited urogynaecology clinic with lower urinary tract symptoms. Each method had low positive predictive values for high post-void residuals (subjective symptom 0.16, uroflowmetry 0.29, pressure flow study 0.28). The prevalence of voiding dysfunction is fairly high and none of the questionnaire and objective bladder emptying tests is sufficient for the diagnosis of urinary retention.

Clinical and genetic analysis of Korean patients with Marfan syndrome: possible ethnic differences in clinical manifestation.

Marfan syndrome (MFS) is an autosomal dominant disorder of the fibrous connective tissue caused by mutations in the fibrillin-1 (FBN1) gene. Although clinical and genetic analyses have been performed in various populations, there have been few studies in Korea. The aim of this study was to investigate the clinical characteristics and genetic background of Korean patients with MFS. In 39 Korean patients with MFS who met the Ghent criteria, the most common clinical finding was aortic dilatation and/or dissection (94.9%), whereas only 35.9% of patients had ectopia lentis. The majority of MFS patients had fewer than four of the skeletal findings required to fulfill the major skeletal Ghent criterion for MFS. Only 21% of Korean patients had major skeletal abnormalities and most cases showed only minor skeletal involvement. FBN1 gene mutations were detected in 35 out of 39 patients (89.7%), which is similar to rates presented in the previous reports. These results suggest that some clinical features in Korean patients with MFS differed from those reported in Western MFS patients.

Analysing malaria incidence at the small area level for developing a spatial decision support system: A case study in Kalaburagi, Karnataka, India.

Spatial decision support systems have already proved their value in helping to reduce infectious diseases but to be effective they need to be designed to reflect local circumstances and local data availability. We report the first stage of a project to develop a spatial decision support system for infectious diseases for Karnataka State in India. The focus of this paper is on malaria incidence and we draw on small area data on new cases of malaria analysed in two-monthly time intervals over the period February 2012 to January 2016 for Kalaburagi taluk, a small area in Karnataka. We report the results of data mapping and cluster detection (identifying areas of excess risk) including evaluating the temporal persistence of excess risk and the local conditions with which high counts are statistically associated. We comment on how this work might feed into a practical spatial decision support system.

Optimizing gDNA extraction from fresh frozen meningioma tissue for downstream genetic analysis.

OBJECTIVE: Meningioma is the most common brain tumor. Genetic mutations in meningioma that include deletion of the neurofibromatosis type 2 gene, (NF2), offer diagnostic information on tumor behavior, recurrence and potential response to treatment. Obtaining high-grade genetic material is critical for accurate, sensitive and robust molecular testing. Currently, no standardized procedure exists for extracting gDNA from meningioma, and this problem was addressed in this report. METHOD: This study compared the yield and quality of extracted gDNA from patient meningioma specimens using an optimized phenol chloroform method and two commercial silica column-based extractions kits and tested respective performances as template in qPCR tests and multiplex ligation-dependent probe amplification (MLPA) NF2 screening. RESULTS: Mean gDNA yields were comparable for each method tested; however, phenol chloroform extraction outperformed column-based kits in all other quality assurance metrics examined. Phenol chloroform extracted gDNA was highly pure, and of a higher fragment size species when compared to column prepared gDNA. qPCR of GAPDH, B2MG, and RPL37A housekeeping genes demonstrated variance in cycle thresholds between patient samples was much lower in the phenol chloroform group. Similarly, primer efficiencies were significantly improved in this sample group which translated to a broader qPCR linear dynamic range and much improved qPCR performance at low concentrations of template. MLPA screening identified NF2 gene deletions in 6 of 12 meningioma samples. Inconsistencies in copy number data for NF2 and reference regions of the genome were observed between gDNA sample extraction groups that included both false negative and positive errors in silica column derived gDNA samples. CONCLUSIONS: This study outlines a highly robust phenol chloroform extraction method for obtaining high-quality gDNA from frozen meningioma tissue and highlights the significance of performing adequate quality assurance when using gDNA for downstream genetic analysis. Most importantly, we demonstrate using gDNA extracted with silica column based kits can lead to diagnostic errors when screening NF2 deletions in meningiomas with MLPA.

Fission yeast hrp1, a chromodomain ATPase, is required for proper chromosome segregation and its overexpression interferes with chromatin condensation.

Hrp1 of Schizosaccharomyces pombe is a member of the CHD protein family, characterized by a chromodomain, a Myb-like telobox-related DNA-binding domain and a SNF2-related helicase/ATPase domain. CHD proteins are thought to be required for modification of the chromatin structure in transcription, but the exact roles of CHD proteins are not known. Here we examine the sub-cellular localization and biochemical activity of Hrp1 and the phenotypes of hrp1 Delta and Hrp1-overexpressing strains. Fluorescence microscopy revealed that Hrp1 protein is targeted to the nucleus. We found that Hrp1 exhibited DNA-dependent ATPase activity, stimulated by both single- and double-stranded DNA. Overexpression of Hrp1 caused slow cell growth accompanied by defective chromosome condensation in anaphase resulting in a 'cut' (celluntimelytorn) phenotype and chromosome loss. The hrp1 Delta mutation also caused abnormal anaphase and mini-chromosome loss phenotypes. Electron micrographs demonstrated that aberrantly shaped nucleoli appeared in Hrp1-overexpressing cells. Therefore, these results suggest that Hrp1 may play a role in mitotic chromosome segregation and maintenance of chromatin structure by utilizing the energy from ATP hydrolysis.

The effect of physician staffing model on patient outcomes in a medical progressive care unit.

PURPOSE: Although evidence supports the impact of intensivist physician staffing in improving intensive care unit (ICU) outcomes, the optimal coverage for progressive care units (PCU) is unknown. We sought to determine how physician staffing models influence outcomes for intermediate care patients. MATERIALS AND METHODS: We conducted a retrospective observational comparison of patients admitted to the medical PCU of an academic hospital during 12-month periods of high-intensity and low-intensity staffing. RESULTS: A total of 318 PCU patients were eligible for inclusion (143 high-intensity and 175 low-intensity). We found that low-intensity patients were more often stepped up from the emergency department and floor, whereas high-intensity patients were ICU transfers (61% vs 42%, P = .001). However, Mortality Probability Model scoring was similar between the 2 groups. In adjusted analysis, there was no association between intensity of staffing and hospital mortality (odds ratio, 0.84; 95% confidence interval, 0.36-1.99; P = .69) or PCU mortality (odds ratio, 0.96; 95% confidence interval, 0.38-2.45; P = .69). There was also no difference in subsequent ICU admission rates or in PCU length of stay. CONCLUSIONS: We found no evidence that high-intensity intensivist physician staffing improves outcomes for intermediate care patients. In a strained critical care system, our study raises questions about the role of the intensivist in the graded care options between intensive and conventional ward care.

Removal ratio of gaseous toluene and xylene transported from air to root zone via the stem by indoor plants.

This work was designed to investigate the removal efficiency as well as the ratios of toluene and xylene transported from air to root zone via the stem and by direct diffusion from the air into the medium. Indoor plants (Schefflera actinophylla and Ficus benghalensis) were placed in a sealed test chamber. Shoot or root zone were sealed with a Teflon bag, and gaseous toluene and xylene were exposed. Removal efficiency of toluene and total xylene (m, p, o) was 13.3 and 7.0 µg·m(-3)·m(-2) leaf area over a 24-h period in S. actinophylla, and was 13.0 and 7.3 µg·m(-3)·m(-2) leaf area in F. benghalensis. Gaseous toluene and xylene in a chamber were absorbed through leaf and transported via the stem, and finally reached to root zone, and also transported by direct diffusion from the air into the medium. Toluene and xylene transported via the stem was decreased with time after exposure. Xylene transported via the stem was higher than that by direct diffusion from the air into the medium over a 24-h period. The ratios of toluene transported via the stem versus direct diffusion from the air into the medium were 46.3 and 53.7% in S. actinophylla, and 46.9 and 53.1% in F. benghalensis, for an average of 47 and 53% for both species. The ratios of m,p-xylene transported over 3 to 9 h via the stem versus direct diffusion from the air into the medium was 58.5 and 41.5% in S. actinophylla, and 60.7 and 39.3% in F. benghalensis, for an average of 60 and 40% for both species, whereas the ratios of o-xylene transported via the stem versus direct diffusion from the air into the medium were 61 and 39%. Both S. actinophylla and F. benghalensis removed toluene and xylene from the air. The ratios of toluene and xylene transported from air to root zone via the stem were 47 and 60 %, respectively. This result suggests that root zone is a significant contributor to gaseous toluene and xylene removal, and transported via the stem plays an important role in this process.

Factors predicting outcomes of penile rehabilitation with udenafil 50 mg following radical prostatectomy.

Udenafil is a selective phosphodiesterase type 5 inhibitor made available in recent years for the treatment of erectile dysfunction. Herein, we evaluated independent predictors of potency recovery in radical prostatectomy (RP) patients who underwent penile rehabilitation with udenafil 50 mg. One hundred and forty-three men who underwent RP were enrolled in a penile rehabilitation program using udenafil 50 mg every other day. The rate of regained potency in the study group was significantly higher compared with the recovery rate seen in patients who were not part of the penile rehabilitation program (41.3% vs 13.0%; P<0.001). On the multivariate Cox analyses, preoperative International Index of Erectile Function-5 scores (hazard ratio (HR), 1.049; P=0.040), alcohol consumption (HR, 2.043; P=0.020) and Gleason biopsy score (HR, 0.368; P=0.024) were independent preoperative predictors for potency recovery. Among post-RP variables, the use of robotic procedures (HR, 2.287; P=0.030) and pathologic stage (HR, 0.506; P=0.038) were significantly associated with potency recovery. This study identified predictive factors for the recovery of potency in patients undergoing penile rehabilitation with udenafil following RP. Our results could provide physicians with useful information for counseling RP patients and selecting optimal candidates for penile rehabilitation.

Photoprotective Effect of Carpomitra costata Extract against Ultraviolet B-Induced Oxidative Damage in Human Keratinocytes.

Natural marine products show various biological properties such as antiphotoaging, antioxidant, anticancer, and anti-inflammation. This study evaluated the protective effects of the brown alga Carpomitra costata (Stackhouse) Batters (Sporochnaceae) against ultraviolet B (UVB)-provoked damage in human HaCaT keratinocytes. C. costata extract (CCE) effectively reduced superoxide anion, hydroxyl radical, and UVB-stimulated intracellular reactive oxygen species (ROS) levels. CCE also restored the expression and activity of UVB-suppressed antioxidant enzymes. Furthermore, CCE decreased UVB-triggered oxidative damage to cellular components including DNA, protein, and lipid and defended the cells against mitochondrial membrane depolarization-medicated apoptosis. The results of this study indicate that CCE can safeguard human keratinocytes against UVB-induced cellular damage via a potent antioxidant mechanism. CCE may find utility as part of a therapeutic arsenal against the damaging effects of UVB radiation on the skin.

A prospective randomized study on the mobilization of CD34+ cells comparing continuous intravenous vs subcutaneous administration of rhG-CSF in normal donors.

The efficacy of mobilizing peripheral blood progenitor cells (PBPC) with continuous intravenous (c.i.v.) administration of rhG-CSF was randomly compared to subcutaneous (s.c.) administration, in 15 normal donors in each arm of the study for 6 days. The percentage and absolute numbers of CD34+ cells in the c.i.v. and s.c. groups increased maximally at day 3 and 5, respectively, when compared with the steady-state (day 0) level. Peak CD34+ cell levels were achieved on day 3 in the c.i.v. group, with more rapid results than in the s.c. group (49.3/microl vs 35.9/microl, P=0.043). Plasma rhG-CSF levels declined progressively during mobilization in each group as the WBC increased. The serum level of rhG-CSF did not correlate with CD34+ cell counts in the peripheral blood. Toxicity profiles in the c.i.v. and s.c. groups were similar. Each regimen was effective in successfully mobilizing the target CD34 cell number.

Expression of vascular endothelial growth factor and microvessel density in head and neck tumorigenesis.

Angiogenesis is a fundamental process in tumor growth and metastasis, and its significance and that of vascular endothelial growth factor (VEGF) expression as prognostic indicators have been documented for various types of human tumors. However, the mechanisms responsible for angiogenesis in head and neck squamous cell carcinoma are not well defined. To examine the relationship between angiogenesis and the phenotypic progressions of head and neck tumorigenesis, we used immunohistochemistry to analyze VEGF expression and microvessel density in 70 paraffin-embedded specimens that contained adjacent normal epithelium, premalignant lesions, or both from 57 patients with head and neck squamous cell carcinoma. Ten samples of normal oral mucosa were obtained from people who did not smoke or drink alcohol and included in the analysis as normal controls. Microvessel density was evaluated by averaging 10 microscopic fields (x400) in a defined area of each specimen. The degree of VEGF expression was assessed on a cell-by-cell basis in 10 microscopic fields (x200) in a defined area on a scale ranging from 0 (no expression) to 3+ (highest level of expression). In addition, the weighted mean index of VEGF expression was calculated. The mean +/- SD weighted mean index of VEGF expression in normal control epithelium (1.10 +/- 0.38, n = 10) was higher than it was in adjacent normal epithelium (0.82 +/- 0.27, n = 13; P = 0.04). VEGF expression decreased as samples ranged from normal adjacent epithelium to hyperplasia (0.78 +/- 0.28, n = 21), mild dysplasia (0.70 +/- 0.29, n = 28), moderate dysplasia (0.67 +/- 0.29, n = 11), severe dysplasia (0.51 +/- 0.39, n = 6), and squamous cell carcinoma (0.20 +/- 0.27, n = 70; overall P = 0.0001). VEGF expression was two times lower in cases with nodal disease (0.17 +/- 0.26, n = 29) than it was in nonnodal disease (0.32 +/- 0.29, n = 16; P = 0.02). Microvessel density showed no significant difference from adjacent normal epithelium premalignant lesions to cancer. In tumor, no correlation was seen between VEGF expression or microvessel density and differentiation, primary tumor site, T stage, or smoking status. These findings indicate that VEGF expression is down-regulated during head and neck tumorigenesis. However, further studies are required to better understand the mechanism of VEGF down-regulation in head and neck tumorigenesis.

Influence of Incorporated Pt-Fe2O3 Core-Shell Nanoparticles on the Resistive Switching Characteristics of ZnO Thin Film.

The resistance-switching characteristics of metal oxides have attracted great interest for the non-volatile memory applications such as resistive random access memory. A basic resistive random access memory device has a metal/insulator/metal structure, and its memory effect is achieved by applying voltage to change the resistance of the insulating layer. One of the promising candidates for explaining the resistance-switching mechanism is the formation and rupture of nanoscale conductive filaments. However, this model has an issue that needs to be addressed: the wide distribution of switching voltage due to randomly formed filaments. Therefore, some researchers have reported a decrease in switching voltage distribution and an increase in switching stability by incorporating nanoparticles into the insulating layer. In this study, we investigated influence of incorporated Pt-Fe2O3 core-shell nanoparticles on the resistive switching characteristics of ZnO thin films. Devices were fabricated on SiO2 wafers. A 100-nm-thick Cr layer was used as the bottom electrode. A 50-nm-thick ZnO layer was deposited using the sputtering method, and Pt-Fe2O3 nanoparticles were deposited on it by the dip coating method. A 50-nm-thick ZnO layer was then deposited again. A top Cr electrode (size: 100 µm x 100 µm) was deposited using a shadow mask and sputtering system. All the devices showed bipolar resistance-switching behavior that is observed in Cr/ZnO/Cr structures. However, the on/off voltage was dramatically lowered by incorporating nanoparticles into the insulating layer when compared with that of the devices without nanoparticles. In addition, the switching stability of the devices was improved upon the incorporation of nanoparticles. On the basis of these results, we can conclude that Pt-Fe2O3 nanoparticles may be used to enhance the resistance switching properties of ZnO thin films by incorporating them into the films.

Targeting the glucose-regulated protein-78 abrogates Pten-null driven AKT activation and endometrioid tumorigenesis.

Rates of the most common gynecologic cancer, endometrioid adenocarcinoma (EAC), continue to rise, mirroring the global epidemic of obesity, a well-known EAC risk factor. Thus, identifying novel molecular targets to prevent and/or mitigate EAC is imperative. The prevalent Type 1 EAC commonly harbors loss of the tumor suppressor, Pten, leading to AKT activation. The major endoplasmic reticulum (ER) chaperone, GRP78, is a potent pro-survival protein to maintain ER homeostasis, and as a cell surface protein, is known to regulate the phosphatidylinositol 3-kinase (PI3K)/AKT pathway. To determine whether targeting GRP78 could suppress EAC development, we created a conditional knockout mouse model using progesterone receptor-Cre-recombinase to achieve Pten and Grp78 (cPten(f/f)Grp78(f/f)) deletion in the endometrial epithelium. Mice with a single Pten (cPten(f/f)) deletion developed well-differentiated EAC by 4 weeks. In contrast, no cPten(f/f)Grp78(f/f) mice developed EAC, even after more than 8 months of observation. Histologic examination of uteri from cPten(f/f)Grp78(f/f) mice also revealed no complex atypical hyperplasia, a well-established EAC precursor. These histologic observations among the cPten(f/f)Grp78(f/f) murine uteri also corresponded to abrogation of AKT activation within the endometrium. We further observed that GRP78 co-localized with activated AKT on the surface of EAC, thus providing an opportunity for therapeutic targeting. Consistent with previous findings that cell surface GRP78 is an upstream regulator of PI3K/AKT signaling, we show here that in vivo short-term systemic treatment with a highly specific monoclonal antibody against GRP78 suppressed AKT activation and increased apoptosis in the cPten(f/f) tumors. Collectively, these findings present GRP78-targeting therapy as an efficacious therapeutic option for EAC.

Apoptosis induction of ultraviolet light A and photochemotherapy in cutaneous T-cell Lymphoma: relevance to mechanism of therapeutic action.

The anti-tumor action of many chemotherapeutic agents has recently been attributed to the induction of apoptosis in the malignant cell population. In this study, we investigated the ability of extracorporeal photopheresis (ExP) and in vitro PUVA (8-methoxy-psoralen + ultraviolet A) therapy to induce apoptosis in peripheral blood mononuclear cells from Sezary syndrome patients and normal controls. Flow cytometric analysis of ExP- or PUVA-treated peripheral blood lymphocytes demonstrated two distinct cell populations within 24 h of treatment. One population was similar to untreated controls with the other exhibiting characteristics of apoptotic cell death, i.e., a loss of cell volume and an accompanying increase in cell density. This latter population was comprised of cells with DNA strand breaks as determined by the Tdt-mediated deoxyuridine triphosphate-biotin nick end labeling assay. Apoptosis was also confirmed morphologically by fluorescent and electron microscopy as well as by demonstration of characteristic DNA strand breaks (laddering) using gel electrophoresis. Apoptosis was not observed with 8-methoxypsoralen (< or = 300 ng per ml) alone; however, ultraviolet A alone at doses > or = 2 J per cm2 induced apoptosis in lymphocytes. Peripheral blood T-cell subpopulations of Sezary syndrome patients, including the malignant clone, were equally susceptible to apoptosis subsequent to either photopheresis or PUVA treatment. In contrast, monocytes (CD14+/CD45+) appear to be resistant to apoptosis induction by ExP or PUVA treatment. Moreover, ExP-treated and untreated monocytes phagocytized apoptotic, but not untreated, peripheral blood mononuclear cells. ExP and PUVA have been shown to be efficacious and well-tolerated therapies in the treatment of dermatologic diseases and transplant rejection. These data suggest that induction of apoptosis may be an important event for therapeutic efficacy.