Various polyunsaturated fatty acids, especially gamma-linolenic acid (GLA), inhibit the growth of a variety of tumor cells. Some evidence indicates that polyunsaturated fatty acid can kill cells by apoptosis. In the current study, we tested the apoptotic effect of GLA on human chronic myelogenous leukemia K562 cells. GLA induced K562 cell death in a dose-dependent manner. Typical apoptotic nuclei were shown by staining of K562 cells with DNA-binding fluorochrome Hoechst 33342, characterized by chromatin condensation and nuclear fragmentation. Flow cytometric analysis also demonstrated that GLA caused dose-dependent apoptosis of K562 cells. The apoptosis could be inhibited by a pancaspase inhibitor (z-VAD-fmk), suggesting the involvement of caspases. Further, release of cytochrome c, activation of caspase-3 and cleavage of PARP were found in GLA-induced apoptosis. GLA treatment could also elevate lipid peroxidation in K562 cells, and antioxidant alpha-tocopherol could reverse the cytotoxicity of GLA. The saturated fatty acid SA, which did not exhibit significant increase in lipid peroxidation, also did not induce cytotoxicity. Intracellular GSH was also determined, and there was no marked change of GSH levels in cells after incubation with GLA compared with the control. These results demonstrate that GLA could induce apoptosis in K562 cells. Apoptosis is mediated by release of cytochrome c, activation of caspase-3. Lipid peroxidation may play a role in GLA cytotoxicity.

OBJECTIVE

This study examines the associations of Internet addiction with social anxiety, depression, and psychosocial well-being among Asian adolescents. A self-medication model conceptualizing Internet addiction as a mediating role in relating depression and social anxiety to negative psychosocial well-being was tested.

METHODS

A cross-sectional survey.

METHODS

In the Asian Adolescent Risk Behavior Survey (AARBS), 5366 adolescents aged 12-18 years from six Asian countries (China, Hong Kong, Japan, South Korea, Malaysia, and Philippines) completed a questionnaire with items of the Internet Addiction Test (IAT), Social Anxiety Scale for Adolescents (SAS-A), Center for Epidemiological Studies Depression Scale (CESD), Self-Rated Health of the Nation Outcome Scales for Children and Adolescents (HoNOSCA-SR) in the 2012-2013 school year. Structural equation modelling was used to examine the mediating role of Internet addiction in depression, social anxiety, and subjective psychosocial well-being.

RESULTS

Significant differences on the scores of IAT, SAS-A, CESD, and HoNOSCA-SR across the six countries were found. The proposed self-medication model of Internet addiction received satisfactory goodness-of-fit with data of all countries. After the path from social anxiety to Internet addiction had been discarded in the revised model, there was a significant improvement of the goodness-of-fit in the models for Japan, South Korea, and the Philippines.

CONCLUSIONS

Depression and social anxiety reciprocally influenced, whereas depression associated with poorer psychosocial well-being directly and indirectly through Internet addiction in all six countries. Internet addiction mediated the association between social anxiety and poor psychosocial well-being in China, Hong Kong, and Malaysia.

The management of symptoms experienced by patients receiving cytotoxic chemotherapy influences quality of life during treatment. Symptom management may be improved through a structured approach to symptom assessment. This paper describes the development of the Chemotherapy Symptom Assessment Scale (C-SAS), a 24-item scale designed for the routine assessment of symptoms experienced by patients receiving cytotoxic chemotherapy. The scale development process focused both upon the psychometric properties and the clinical usefulness of the scale. Patients and health professionals played a significant role in item selection and scale design in order to maximise the clinical utility of the C-SAS.

This study evaluated the psychometric properties of the Chinese version of the Social Anxiety Scale for Adolescents (SAS-A) in a sample of 296 adolescents (49% boys) in Grades 7, 8, 9, 10, and 12 with a mean age of 15.52 years. Confirmatory factor analysis replicated the three-factor structure of the SAS-A in the Chinese sample: Fear of Negative Evaluation (FNE), Social Avoidance and Distress in New Situations (SAD-New), and Social Avoidance and Distress-General (SAD-General). Internal consistency and test-retest reliability were appropriate. The results also revealed a clear and predictable pattern of relationships between the SAS-A and the Questionnaire about Interpersonal Difficulties for Adolescents and the International Personality Item Pool. Chinese boys reported greater SAD-General than Chinese girls, and this difference increased with grade. The SAS-A scores were compared to previously collected data from the USA and Spain, revealing that Chinese adolescents scored significantly higher in social anxiety than American and Spanish adolescents.

Cadmium (Cd) is a widespread toxic heavy metal that usually causes deleterious effects on plant growth and development. Salicylic acid (SA), a naturally existing phenolic compound, is involved in specific responses to various environmental stresses. To explore the role of SA in the tolerance of melon (Cucumis melo L.) to Cd stress, the influence of SA application on the growth and physiological processes was compared in the two melon cultivars Hamilv (Cd-tolerant) and Xiulv (Cd-sensitive) under Cd stress. Under 400-μM Cd treatment, Hamilv showed a higher biomass accumulation, more chlorophyll (Chl), greater photosynthesis, and less oxidative damage compared to Xiulv. Foliar spraying of 0.1 mM SA dramatically alleviated Cd-induced growth inhibition in the two melon genotypes. Simultaneously, SA pretreatment attenuated the decrease in Chl content, photosynthetic capacity, and PSII photochemistry efficiency in Cd-stressed plants. Furthermore, exogenous SA significantly reduced superoxide anion production and lipid peroxidation, followed by increase in the activities of antioxidant enzyme superoxide dismutase, guaiacol peroxidase, catalase, and ascorbate peroxidase, and content of soluble protein and free proline in both the genotypes under Cd stress. The effect of SA was more conspicuous in Xiulv than Hamilv, reflected in the biomass, photosynthetic pigments, stomatal conductance, water use efficiency, and antioxidant enzymes. These results suggest that exogenous spray of SA can alleviate the adverse effects of Cd on the growth and photosynthesis of both the melon cultivars, mostly through promoting antioxidant defense capacity. It also indicates that SA-included protection against Cd damage is to a greater extent more pronounced in Cd-sensitive genotype than Cd-tolerant genotype.

Age-related osteoporosis is characterized by the deterioration in bone volume and strength, partly due to the dysfunction of bone marrow mesenchymal stromal/stem cells (MSCs) during aging. Alpha-ketoglutarate (αKG) is an essential intermediate in the tricarboxylic acid (TCA) cycle. Studies have revealed that αKG extends the lifespan of worms and maintains the pluripotency of embryonic stem cells (ESCs). Here, we show that the administration of αKG increases the bone mass of aged mice, attenuates age-related bone loss, and accelerates bone regeneration of aged rodents. αKG ameliorates the senescence-associated (SA) phenotypes of bone marrow MSCs derived from aged mice, as well as promoting their proliferation, colony formation, migration, and osteogenic potential. Mechanistically, αKG decreases the accumulations of H3K9me3 and H3K27me3, and subsequently upregulates BMP signaling and Nanog expression. Collectively, our findings illuminate the role of αKG in rejuvenating MSCs and ameliorating age-related osteoporosis, with a promising therapeutic potential in age-related diseases.

Alpha-globin gene cluster haplotypes were determined in Southern African San and negroid populations. Significant differences (P less than .01) between the two groups were found at three of the nine loci in the cluster. The most striking difference, however, was the relatively low level of variation found in the San (alpha alpha)-associated haplotypes and the high level in the SA blacks. This trend was also observed for the 3' hyper-variable region. Nineteen different haplotypes were identified among the 36 haplotypes studied in the black population, but only seven different ones were found among the 37 haplotypes in the San; five were common to both populations. The common San haplotype, (+--MPZ+---), had a frequency of .57 in the San and .11 in the black population; the common SA black haplotype, (---MZ----), occurred at a frequency of .17 but was absent in the San. In the SA black population significant linkage disequilibrium is present between five of the RFLP loci, including the extreme 5' and 3' markers, confirming the absence of a recombination hot spot in the alpha-globin gene cluster.

The role of MHC class II antigens was investigated in the process of antigen binding by T8+ cells and monocytes (Mo) and in the functions of helper factor (HF) and suppressor factor (SF). Monoclonal antibodies (MoAbs) to HLA-DR, DC and SB determinants were used in immunofluorescence, inhibition of antigen binding and affinity chromatography of HF and SF. Indirect immunofluorescence studies suggest that T lymphocytes from peripheral blood of healthy subjects have a small proportion of cells expressing HLA-DR, beta chain determinants (1.4-3.8%). These belong predominantly to the T8+ subset of cells (4.6-8.8%), with only a very small proportion in the T4+ cells (0.1-1.8%). However, DC1 on DRw6+ T cells and SB2,3 on any HLA typed cells were found in significantly greater proportion than the DR antigens in both T8+ and T4+ cells, though this was again greater on T8+ (30 and 25%) than T4+ (8.3 and 14.4%) cells. Although Mo had a greatly increased proportion of cells with DR-beta chain determinants (27-45%) than the T8+ cells, the converse was found with DC1 and SB2,3 determinants (13.9 and 11.4%). Inhibition of 125I-streptococcal antigen (SA) binding to T8+ cells and to Mo by MoAbs to the class II antigens showed that DR-beta chain monomorphic or polymorphic antibodies and DC1 antibodies inhibited binding to both cell types by 66-94%. However, MoAbs to DR-alpha chains or to the SB2,3 determinant failed to yield significant inhibition. Affinity chromatography studies of HF and SF revealed that the DR-beta chain monomorphic and DC1 antibodies bound HF and SF activities and that this was not found with the DR-beta chain polymorphic or SB2,3 antibodies. The results of inhibition of 125I-SA binding to T8+ cells and Mo, and absorption of HF and SF by affinity chromatography with MoAbs suggest four categories of recognition of human MHC class II antigenic determinants. (1) Class II determinants shared by the T8+ cells, Mo, HF and SF and recognized by MoAbs to monomorphic beta chains (DA6.231) and to DC1. (2) Class II determinants shared only by the SA binding T8+ cells and Mo and recognized by the MoAbs to a polymorphic beta chain (DA6.164) and to a monomorphic DR determinant (OK.Ial). (3) Class II determinants shared only by the HF and SF and recognized by the MoAbs to one of the alpha chains (TAL.1B5). (4) Class II determinants not detected on the two cells or the two T cell factors.

The present investigation examined the effects of placebo (P), low dose (LD) and high dose (HD) ethanol on EEG activity in two groups of males. One group consisted of individuals at high risk for the development of alcoholism (HR, N = 21) while the other consisted of matched, low risk (LR, N = 21) controls. Only one condition (P, LD or HD) was presented each day and condition order was randomized. For each subject, both blood alcohol level(s) (BAL) measured via breathalyzer and EEG activity, using the entire 10/20 international system, were recorded prior to and at intervals of 35, 70, 105 and 140 min after P, LD or HD administration. The Fast Fourier Transform (FFT) was used to calculate power spectral densities (PSD). Measures of relative area under the power spectral curve were obtained for each of the following frequency bands: slow alpha (SA, 7.5-10 Hz), fast alpha (FA, 10.5-13.0 Hz), slow beta (SB, 13.5-19.5 Hz) and fast beta (FB, 20-26 Hz) at electrodes: F3, F4, C3, C4, P3, P4, O1 and O2. The results of repeated measures MANOVA conducted on the normalized values of relative areas revealed that at each electrode examined, ethanol elicited significant changes only in SA activity. Risk group differences in SA activity were observed only at electrodes F3, F4 and P4. These differences were the consequence of differential ethanol effects rather than differences in baseline SA levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Profiling of carbohydrate structures on cell membranes has been difficult to perform because of the complexity and the variations of such structures on cell surface glycans. This study presents a novel method for rapid profiling of cell surface glycans for terminal N-acetyllactosamines (Galbeta1-(3)4GlcNAc-R) that are uncapped, capped with sialic acid as SA-Galbeta1-(3)4GlcNAc-R, or with alphaalpha-gal epitope- Galalphaalpha-gal epitopes are synthesized on the exposed N-acetyllactosamines by alphaalpha-gal epitopes on cells are quantified by a modification of radioimmunoassay designated as "ELISA inhibition assay," which measures binding of the monoclonal anti-Gal antibody M86 to alpha-gal epitopes. This binding is proportional to the number of cell surface alpha-gal epitopes. The amount of free M86 antibody molecules remaining in the solution is determined by ELISA using synthetic alpha-gal epitopes linked to albumin as solid phase antigen. The number of alpha-gal epitopes on cells is estimated by comparing binding curves of M86 incubated with the assayed cells, at various concentrations of the cells, with the binding of M86 to rabbit red cells expressing 2 x 10(6) alpha-gal epitopes/cell. We could demonstrate large variations in the number of sialic acid capped N-acetyllactosamines, alpha-gal epitopes and uncapped N-acetyllactosamines on different mammalian red blood cells, and on nucleated cells originating from a given tissue in various species. This method may be useful for rapid identification of changes in glycosylation patterns in cells subjected to various treatments, or in various states of differentiation.

Several studies have been performed to examine the problem of diagnosing gastroduodenal reflux (GDR). No single method is widely accepted. The aim of this work was to evaluate the diagnostic value of gastric pHmetry in this regard. A gastric aspiration probe attached to a combined glass electrode was placed in the stomach of 24 patients, with its distal tip located between 9 and 12 cm below the cardia. One ml samples of gastric juice were taken from 8 of the patients every 30 min for 15 h and as well as, every time a spontaneous alkalinization (SA) (defined by a pH greater than or equal to 4 for at least 1 min) was observed. The pH of each sample was measured by colorimetry whereas the concentration of total biliary acids (CTBA) was evaluated by the fluorimetric method (Kit Sterognost 3 alpha Flu); pH value measured via the intragastric electrode during aspiration was also recorded (protocol A). Continuous gastric aspiration was carried out in the remaining 16 patients for the entire duration of the test (6 h) which was divided into periods of 20 min. Apart from the parameters evaluated during protocol A, the percentage of time during which the stomach had a pH greater than or equal to 4 was recorded, as well as the quantity of total biliary acids collected over the 20 min periods (protocol B). Correlation studies were carried out using the Kendall tau and Spearman tests. Percentages were compared using the chi 2 test.(ABSTRACT TRUNCATED AT 250 WORDS)

Drug addiction is associated with impaired judgment in unstructured situations in which success depends on self-regulation of behavior according to internal goals (adaptive decision-making). However most executive measures are aimed at assessing decision-making in structured scenarios, in which success is determined by external criteria inherent to the situation (veridical decision-making). The aim of this study was to examine the performance of Substance Abusers (SA, n = 97) and Healthy Comparison participants (HC, n = 81) in two behavioral tasks that mimic the uncertainty inherent in real-life decision-making: the Cognitive Bias Task (CB) and the Iowa Gambling Task (IGT) (administered only to SA). A related goal was to study the interdependence between performances on both tasks. We conducted univariate analyses of variance (ANOVAs) to contrast the decision-making performance of both groups; and used correlation analyses to study the relationship between both tasks. SA showed a marked context-independent decision-making strategy on the CB's adaptive condition, but no differences were found on the veridical conditions in a subsample of SA (n = 34) and HC (n = 22). A high percentage of SA (75%) also showed impaired performance on the IGT. Both tasks were only correlated when no impaired participants were selected. Results indicate that SA show abnormal decision-making performance in unstructured situations, but not in veridical situations.

OBJECTIVE

To explore ambient PM2.5 the influence of the inflammation injury and the on immune function.

METHODS

The model rats were administered with PM2.5 by the intratracheal instillation. The pathological varieties of rat's lungs and other important organs were observed by light microscope. The protein and SA levels in bronchoalveolar lavage fluids (BALF) were detected by relevant kits. The TNF-alpha, and IL-6 levels in the BALF were detected by ELISA, and the mRNA expression levels in the lung tissue were observed by RT-PCR. The AM were collected to detect the phogacytic function. The ConA-stimulated T lymphocyte proliferation tests were performed to research the proliferation of spleen.

RESULTS

Foreign-body granulomas were observed in the lungs of exposed rats. Monocytes-macrophages assembling in blood sinus of liver and the trend to form the granulomas were observed. Alveolar macrophages containing PM2.5 and dissociative PM2.5 were obviously observed in lung visceral pleural lymphatics and the blood vessels in lung and kidney. The number of the granulomas in the lungs of the rats become more and more as times goes on. The concentrations of total protein and SA in BALF were increased with the dose and time of exposure. TNF-alpha levels in the BALF increased by the dose and exposure time during 3 months, but TNF-alpha levels in the BALF decreased significantly on the 6th month. The expression levels of IL-6 in the BALF increased by the dose. It showed the dose-response relationship. The highest expression level of IL-6 was detected on the 3rd month. The expression level of IL-6 decreased on the 6th month. Phagocytiosis functions of AM were impaired by PM2.5 , which may in turn impair the nonspecific defenses function of airway. But the splenic lymphocyte proliferation were not obviously changed.

CONCLUSIONS

The persistent inflammatory injury was induced by the subchronic exposure to PM2.5. The injury of immunological system were increased with the dose and the time of the exposure. The cytokine net was disordered by PM2.5, which worsen the injury. The phagocytiosis function of AM was impaired by PM2.5, which may be the mechanism of chronic pulmonary diseases.

The controlled release of salicylic acid (SA), a key phytohormone, was mediated by using a novel decanethiol gatekeeper system grafted onto mesoporous silica nanoparticles (MSNs). The decanethiol was conjugated only to the external surfaces of the MSNs through glutathione (GSH)-cleavable disulfide linkages and the introduction of a process to assemble gatekeepers only on the outer surface so that the mesopore area can be maintained for high cargo loading. Raman and nitrogen sorption isotherm analyses confirmed the successful linkage of decanethiol to the surface of MSNs. The in vitro release of SA from decanethiol gated MSNs indicated that the release rate of SA in an environment with a certain amount of GSH was significantly higher than that without GSH. More importantly, in planta experiments showed the release of SA from decanethiol gated MSNs by GSH induced sustained expression of the plant defense gene PR-1 up to 7 days after introduction, while free SA caused an early peak in PR-1 expression which steadily decreased after 3 days. This study demonstrates the redox-responsive release of a phytohormone in vitro and also indicates the potential use of MSNs in planta as a controlled agrochemical delivery system.

<Abstr<em>a</em>ctText>Crosst<em>a</em>lk between c<em>a</em>ncer cells <em>a</em>nd c<em>a</em>rcinom<em>a</em>-<em>a</em>ssoci<em>a</em>ted fibrobl<em>a</em>sts (CAFs) is known to be involved in v<em>a</em>rious <em>a</em>spects of tumor biology, including during inv<em>a</em>sion. Using or<em>a</em>l squ<em>a</em>mous cell c<em>a</em>rcinom<em>a</em> (OSCC) cells <em>a</em>s <em>a</em> model, we ex<em>a</em>mined whether <em>a</em>nd how CAFs respond to infl<em>a</em>mm<em>a</em>tory sign<em>a</em>ls to influence c<em>a</em>ncer cell migr<em>a</em>tion <em>a</em>nd inv<em>a</em>sion.</Abstr<em>a</em>ctText><Abstr<em>a</em>ctText>Chemokine sign<em>a</em>tures within the hum<em>a</em>n HNSCC d<em>a</em>t<em>a</em>sets from The C<em>a</em>ncer Genome Atl<em>a</em>s (TCGA) were <em>a</em>n<em>a</em>lyzed together with tissue <em>a</em>ssessment using immunohistochemic<em>a</em>l st<em>a</em>ining (IHC) <em>a</em>nd re<em>a</em>l-time PCR. A co-culture system w<em>a</em>s used to identify reciproc<em>a</em>l effects exerted by CAFs <em>a</em>nd c<em>a</em>ncer cells upon one <em>a</em>nother. Recombin<em>a</em>nt CXCL1, CXCL1 neutr<em>a</em>lizing <em>a</em>ntibodies, <em>a</em>nd CXCR2 <em>a</em>nt<em>a</em>gonist were used to confirm CXCL1/CXCR2 <em>a</em>xis-medi<em>a</em>ted cell beh<em>a</em>viors.</Abstr<em>a</em>ctText><Abstr<em>a</em>ctText>An<em>a</em>lysis of the TCGA d<em>a</em>t<em>a</em>set reve<em>a</em>led th<em>a</em>t CXCL1 is <em>a</em>ssoci<em>a</em>ted with poor surviv<em>a</em>l, <em>a</em>nd IHC demonstr<em>a</em>ted CXCL1 is highly expressed in OSCC strom<em>a</em>l cells. Moreover, re<em>a</em>l-time PCR showed th<em>a</em>t in <em>a</em>ddition to CXCL1, IL-1β <em>a</em>nd CXCR2 <em>a</em>re <em>a</em>lso highly expressed in OSCC <em>a</em>nd IL-1β mRNA levels positively correl<em>a</em>te with CXCL1 expression. Furthermore, CAFs co-cultured with <em>SAS</em>, <em>a</em> poorly differenti<em>a</em>ted OSCC cell line, or stimul<em>a</em>ted with IL-1β exhibit incre<em>a</em>sed CXCL1 secretion in <em>a</em>n NF-κB-dependent m<em>a</em>nner. Tre<em>a</em>tment of <em>SAS</em> cells with CAF-conditioned medium or CXCL1 incre<em>a</em>sed their inv<em>a</em>sion <em>a</em>nd migr<em>a</em>tion c<em>a</em>p<em>a</em>bilities, indic<em>a</em>ting <em>a</em> reciproc<em>a</em>l <em>a</em>ctiv<em>a</em>tion between CAFs <em>a</em>nd c<em>a</em>ncer cells. Moreover, CXCL-1 upregul<em>a</em>ted m<em>a</em>trix met<em>a</em>lloprote<em>a</em>se-1 (MMP-1) expression <em>a</em>nd <em>a</em>ctivity in CAFs.</Abstr<em>a</em>ctText><Abstr<em>a</em>ctText>The induction of IL-1β following CXCL1 stimul<em>a</em>tion of CAFs medi<em>a</em>tes c<em>a</em>ncer cell inv<em>a</em>sion, <em>a</em>nd there is <em>a</em> reciproc<em>a</em>l dependency between CAFs <em>a</em>nd c<em>a</em>ncer cells in the OSCC microenvironment.</Abstr<em>a</em>ctText>

Docosahexaenoic (DHA; C22:6 n-3), eicosapentaenoic (EPA; C20:5 n-3), palmitic (PA; C16:0), and stearic (SA; C18:0) acids decrease lymphocyte proliferation in concentrations of >50 muM, as observed in our previous study. However, oleic acid (OA; C18:1 n-9) and linoleic acid (LA; C18:2 n-6) increase lymphocyte proliferation at 25 muM. In this study, the effect of these FAs on the interleukin-2 (IL-2) signaling pathway in human lymphocytes was investigated. Cells were isolated from heparinized venous blood of healthy human donors by density-gradient sedimentation. Cells were stimulated with 5 mug/ml concanavalin A and treated with FAs in the absence or presence of IL-2 for 1 hour. CD25-alpha externalization was analyzed by flow cytometry, and Janus kinase 1 (JAK1), JAK3, signal transducer and activator of transcription (STAT) 5, extracellular signal-regulated kinases (ERKs) 1 and 2, Akt, and protein kinase C (PKC)-zeta phosphorylation were analyzed by Western blotting. The expression of CD25-alpha at the cell surface was increased by DHA, SA, and PA but was unaffected by EPA, OA, and LA. PA, SA, DHA, and EPA decreased JAK1, JAK3, STAT5, and Akt phosphorylation induced by IL-2, but OA and LA did not cause any effect. OA and LA increased ERK1/2 phosphorylation, whereas the other FAs caused a marked decrease. PKC-zeta phosphorylation was decreased by OA and LA and was not altered by the remaining FAs. In conclusion, the inhibitory effect of PA, SA, DHA, and EPA on lymphocyte proliferation observed in our previous study was attributable to a decrease in JAK/STAT, ERK, and Akt pathways activated by IL-2. Probably, OA and LA stimulated lymphocyte proliferation by increasing ERK1/2 phosphorylation through PKC-zeta activation. The inhibition of JAK1, JAK3, STAT5, ERK1/2, and Akt phosphorylation caused by DHA, SA, and PA is associated with an alteration of CD25 expression at the cell surface.

OBJECTIVE

Retinoic acid (RA) and benzoyl peroxide (BP) were studied, comparing their keratolytic efficacy and water barrier disruption to that of salicylic acid (SA), a well-established keratolytic, under similar conditions.

METHODS

Six volunteers were included in this blinded study. Eleven randomized test sites were marked on the volar forearms, containing sites for untreated skin at time zero, unoccluded, occlusion, and vehicle controls for 3 and 6 h, and each of BP, RA, and SA solutions for 3 and 6 h. At each time point, occlusion at 5 of the test sites was removed, and chromameter measurements were performed over 30 min. Each site then underwent 25 stratum corneum (SC) tape strippings. At 1, 5, and 30 min after the last stripping at each site, TEWL measurements were performed. Quantitative protein analysis of the SC from the tapes was then performed.

CONCLUSIONS

after 3 h, bp was significantly more effective in disrupting sc cohesion than sa and ra, indicating bp is a moderate keratolytic agent in addition to its antimicrobial properties. After 6 h, all three agents were similarly effective in keratolysis. Barrier disruption, as measured by TEWL, paralleled depth of SC removal. SA tended to exhibit the greatest keratolytic efficacy superficially, hence its clinical effectiveness in superficial conditions such as comedonal acne, whereas BP was more effective at deeper levels, complimenting its antimicrobial effects and enabling it to treat deeper, more inflammatory lesions. None of the agents significantly affected skin erythema. These techniques provide a robust and rapid assay for in vivo keratolytic demonstration.

We investigated the antidepressant effects of bilateral intra-the ventrolateral orbital cortex (VLO) administration of sanguinarine (SA), a selective mitogen-activated protein kinase phosphatase-1 (Mkp-1) inhibitor, in rats that had been subjected to a forced swimming test (FST) which is a classic animal model of depression. The expression of Mkp-1 and activation of ERK (ratio of phosphor-ERK to ERK) were also examined by immunoblotting. A single bilateral intra-VLO infusion of SA (2.5, 5 or 10 μg/0.5 μl per side) significantly reduced immobility time in the FST in dose-dependent fashion, as compared to vehicle-treated controls. A similar antidepressant effect was also observed in rats systemically administered fluoxetine, a classic antidepressant. The effects observed in the FST could not be attributed to non-specific increases in activity as neither microinjection of SA into the VLO nor fluoxetine treatment altered the behavior of the rats during the locomotion test. In addition, a decrease in the expression of Mkp-1 and a correlative increase in ERK activation were involved in the antidepressant effects of the bilateral SA administration into the VLO. The results indicated that Mkp-1 within the VLO is involved in the process of depression and may be a potential target for therapeutic action of antidepressant treatment.

Reactive oxygen species (ROS) act as signaling molecules for regulating plant responses to abiotic and biotic stress and there exist source- and kind-specific pathways for ROS signaling. Recently, we created a novel system for producing H 2O 2 in Arabidopsis chloroplasts by chemical-dependent thylakoid membrane-bound ascorbate peroxidase (tAPX) silencing using an estrogen-inducible RNAi method. Microarray analysis revealed that the expression of a large set of genes was altered in response to tAPX silencing, some of which are known to be involved in pathogen response/resistance. Furthermore, we found that tAPX silencing enhances the levels of salicylic acid (SA) and the response to SA, a central regulator for biotic stress response. In this addendum, we describe the relationship between chloroplastic H 2O 2 and SA in stress response, and discuss the function of the kind- and source-specific ROS signaling in SA-mediated stress response.

OBJECTIVE

Circumstantial evidence indicates that, in the presence of a suitable substratum, sudden, behaviorally induced increases in sympathetic drive to the cardiovascular system might play an important physiopathological role in various conditions, ranging from arterial hypertension to sudden coronary death. Accordingly, it might be useful to study the effects of behavioral interventions, such as mental relaxation, that might be capable of blunting excitatory autonomic responses. It would also be preferable to study healthy subjects in whom autonomic control is not modified by the presence of disease, and to use noninvasive approaches to minimize the possible emotional impact produced by invasive recordings.

METHODS

We examined healthy subjects who were either subjected to relaxation training (N = 13) or sham relaxation (N = 12). An additional group, treated with beta-adrenergic blockade (N = 12), was also examined. Spectral and cross-spectral analysis of RR interval and systolic arterial pressure (SAP) variabilities provided quantitative markers of sympathovagal balance modulating the sinoatrial (SA) node, of sympathetic vasomotor modulation, and of the gain of the arterial pressure/heart period baroreflex (index alpha). Subjects were studied at rest, during standing, and during mental arithmetic.

RESULTS

Data indicate that both beta-adrenergic blockade and relaxation training significantly blunted the excitatory autonomic responses to standing and to mental arithmetic. Indices of sympathetic modulation also seemed reduced by beta blockade at rest. No changes were observed with sham training.

CONCLUSIONS

Frequency domain analysis of cardiovascular variabilities, using a totally noninvasive approach, indicates that relaxation training significantly blunts the excitatory autonomic changes produced by standardized behavioral laboratory stimuli.

The molecular nature of tissue-specific gene regulation by androgens has not been well defined, partly as a result of the variable expression and incomplete regulation of currently available gene models. We have therefore aimed to establish more informative models by identifying alternative genes whose expression is tightly and coordinately regulated by androgens. Female C57BL/6 mice were dosed with dihydrotestosterone- or sham-treated for 8 days, after which kidneys were removed and complementary DNA (cDNA) prepared. We then applied the subtractive hybridization techniques of random arbitrarily primed-PCR and PCR-coupled subtractive hybridization method of cDNA representational difference analysis to the isolated cDNA. In addition to well characterized androgen-regulated genes [e.g. KAP (kidney androgen-regulated protein)], we demonstrate the differential expression of six genes previously not known to be under androgen control. RNA levels of SA, Cytochrome P450 4B1, IL-6ST (interleukin-6 signal transducer), OATP (organic anion transporter), and a newly identified gene, MJAM, were up-regulated by androgen, while 16-alpha-hydroxylase was decreased. Expression of these transcripts was inhibited in dihydrotestosterone-treated females by flutamide and in males by castration, confirming their dependence on androgens. Although all the genes demonstrate tissue-specific regulation by androgen, SA showed both kidney specificity and absolute requirement for androgen for its expression. These newly identified androgen-regulated genes will constitute very useful models for studying the nature of tissue-specific gene regulation by androgens.

We compared physiological and ultrastructural indices of acute lung injury (ALI) during septic shock caused by taxonomically diverse pathogens to distinguish ALI due to endogenous inflammatory mediators vs. microbial exotoxins or other factors. Conscious rats were infected i.v. with gram-negative Escherichia coli (EC, serotype 055:B5), exotoxin-C producing gram-positive Staphylococcus aureus (SA), or yeast-phase Candida albicans (CA, a clinical isolate). Viable inocula of 10(10) EC, 10(10) SA, or 10(9) CA caused lethal shock in < 24 h, but distinct types of ALI were noted after bacteria vs. fungi. Within 0.5 h of EC infection, leukocytes marginated in the lung vasculature; by death at 6-14 h, animals were hyperoxemic but not acidemic, and showed slight interstitial edema with increased wet/dry weight ratios (W/D = 5.22 +/- 0.10, mean +/- SE, vs. 4.86 +/- 0.07 in controls, P < 0.05). Similarly mild ALI occurred after 10(10) SA. In contrast, within 0.5 h of CA infection, yeast were visible within lung intravascular leukocytes. By death at 6-12 h, CA animals showed hyperoxic acidemia and moderate ALI with capillary obstruction, interstitial hemorrhage, and elevated lung W/D (5.52 +/- 0.13, P < 0.01 vs. controls) associated with yeast-mycelial transformation. Prior neutropenia accelerated mortality and worsened ALI after CA, with hypoxemic acidemia, increased lung W/D (7.23 +/- 0.34, P < 0.05 vs. other groups), capillary occlusion, perivascular and alveolar hemorrhage, and septal disruption by mycelia. Bacteremia induced large increases in serum tumor necrosis factor-alpha (TNF) and interleukin-1 alpha within 1.5 h, but these cytokines remained low in CA animals, even at death. Neither survival nor ALI after EC or CA was altered by pentoxifylline, which attentuated TNF production, or by cyclooxygenase inhibition with ibuprofen. Thus, overall ALI severity correlated with physiological indices of pulmonary function, but ultrastructural changes correlated better with pathogen type than circulating cytokine or eicosanoid mediators. Whereas lethal bacteremia induced early cytokinemia and mild ALI with or without bacterial exotoxins, moderate ALI apparently was mediated by fungal exotoxins during lethal candidemia, which worsened during neutropenia due to enhanced mycelial proliferation.

The effect of sodium arsenite (SA) on LPS-induced NO production in RAW 267.4 murine macrophage cells was studied. SA pretreatment of LPS-stimulated RAW cells resulted in a striking reduction in NO production. No significant difference in LPS binding was observed between RAW cells pretreated with SA and control untreated RAW cells, suggesting that SA might impair the intracellular signal pathway for NO production. SA inhibited LPS-induced NF-kappaB activation by preventing loss of IkappaB-alpha and -beta. Furthermore, SA blocked phosphorylation of extracellular signal-regulated kinase 1/2 (Erk1/2), but not phosphorylation of p38 and c-Jun N-terminal kinase. SA treatment resulted in the disappearance of Raf-1, suggesting that it might cause the inhibition of the Erk1/2 mitogen-activated protein (MAP) kinase pathway. The SA-mediated loss of Raf-1 also abolished LPS-induced NF-kappaB activation as well as the Erk1/2 pathway. The dominant negative mutant of MAP kinase kinase 1 inhibited both NO production and NF-kappaB activation in LPS-stimulated RAW cells. Taken together, these results indicate that the inhibitory action of SA on NO production in LPS-stimulated macrophages might be due to abrogation of inducible NO synthase induction, and it might be closely related to inactivation of the NF-kappaB and Erk1/2 MAP kinase pathways through loss of Raf-1.

1. The aim of this study was to test whether parabrachial area (PBA) stimulation exerts inhibitory influences on the spontaneous activity and responses evoked by skin and deep afferent inputs in trigeminal subnucleus caudalis (Vc) neurons, and to compare these effects with those of nucleus raphe magnus (NRM) stimulation. A total of 92 nonnociceptive and nociceptive Vc neurons was recorded in urethan/alpha-chloralose-anesthetized rats. Each neuron was functionally classified as low-threshold mechanoceptive (LTM), wide dynamic range (WDR), nociceptive-specific (NS), nociceptive convergent with both skin and deep inputs (S+D), or deep nociceptive (D); the LTM neurons could be subdivided as rapidly adapting (RA) or slowly adapting (SA). Conditioning stimulation was applied to histologically verified sites in PBA and NRM. 2. The spontaneous or evoked activity of all classes of neurons could be inhibited by PBA as well as by NRM stimulation, but generally the incidence and magnitude of inhibition were lower for the LTM neurons. Occasionally, facilitation of neuronal activity was also produced by PBA and NRM stimulation. 3. The spontaneous activity of 11 LTM neurons (6 RA, 5 SA), 13 nociceptive neurons (6 WDR, 7 NS), and 5 D neurons was tested with stimulation of PBA or NRM or both. LTM spontaneous activity was more significantly inhibited by NRM stimulation than by PBA stimulation, whereas both NRM and PBA stimulation had similar and significant inhibitory effects on NS, WDR, and D neurons. 4. The evoked nonnociceptive responses of 28 LTM neurons (16 RA, 12 SA) and of 6 WDR neurons were also tested with stimulation of PBA or NRM or both. The magnitudes of inhibition of the responses produced by PBA conditioning stimulation were statistically significantly less than those induced by NRM conditioning stimulation. 5. The cutaneous and deep nociceptive responses of cutaneous nociceptive neurons (9 NS, 19 WDR) and seven D neurons, respectively, were also tested with PBA and NRM stimulation. There was a significant difference in potency between PBA- and NRM-induced inhibition, but no difference in the magnitude of inhibitory effects among NS, WDR, and D neurons. For both PBA and NRM conditioning stimulation, graded increases in intensities of stimulation produced linear increases in inhibitory effects on nociceptive responses; an increase in stimulation frequency from 5 to 400 Hz also produced increases in inhibition of the nociceptive responses. 6. In five S+D nociceptive convergent neurons, the responses elicited by deep inputs were more powerfully inhibited by PBA stimulation than those elicited by cutaneous inputs.(ABSTRACT TRUNCATED AT 400 WORDS)