Effects of virtual exposure to urban greenways on mental health.

Urban greenways (UGW) are increasingly recognized as vital components of urban green infrastructure (UGI). While existing research has provided empirical evidence on the positive impacts of UGW on physical health, studies focusing on the effects on mental health remain limited. Moreover, previous investigations predominantly compare UGW as a whole with other built environments, neglecting the influence of specific vegetation designs along UGW on mental health. To address this research gap, we conducted a randomized controlled experiment to examine the impact of vegetation design along UGW on stress reduction and attention restoration. A total of 94 participants were randomly assigned to one of four UGW conditions: grassland, shrubs, grassland and trees, or shrubs and trees. Utilizing immersive virtual reality (VR) technology, participants experienced UGW through a 5-min video presentation. We measured participants' subjective and objective stress levels and attentional functioning at three time-points: baseline, pre-video watching, and post-video watching. The experimental procedure lasted approximately 40 minutes. Results of the repeated-measures ANOVA revealed that participants experienced increased stress and mental fatigue after the stressor and decreased levels following the UGW intervention. Furthermore, between-group analyses demonstrated that the shrubs group and the grassland and trees group exhibited significantly greater stress reduction than the grassland group. However, there are no significant differences in attention restoration effects between the four groups. In conclusion, virtual exposure to UGW featuring vegetation on both sides positively affected stress reduction and attention restoration. It is recommended that future UGW construction incorporates diverse vegetation designs, including shrubs or trees, instead of solely relying on grassland. More research is needed to explore the combined effects of shrubs and trees on mental health outcomes.

Impacts of urban green infrastructure on attentional functioning: insights from an fMRI study.

Multiple studies using various measures, technologies, and participant groups have found that exposure to urban green infrastructure can help alleviate the daily attentional fatigue that human experience. Although we have made significant progress in understanding the effects of exposure to urban green infrastructure on attention restoration, two important gaps in our knowledge remain. First, we do not fully understand the neural processes underlying attention restoration that exposure to urban green infrastructure elicits. Second, we are largely unaware of how typical patterns of urban green infrastructure, such as combinations of trees and bioswales, affect recovery from attentional fatigue. This knowledge is crucial to guide the design and management of urban landscapes that effectively facilitate attention restoration. To address these gaps in our knowledge, we conducted a controlled experiment in which 43 participants were randomly assigned to one of three video treatment categories: no green infrastructure (No GI), trees, or trees and bioswales. We assessed attentional functioning using functional Magnetic Resonance Imaging (fMRI) and the Sustained Attention Response Task (SART). Participants exposed to urban settings with trees exhibited improved top-down attentional functioning, as evidenced by both fMRI and SART results. Those exposed to urban settings with trees and bioswales demonstrated some attention-restorative neural activity, but without significant improvements in SART performance. Conversely, participants exposed to videos of urban environments without green infrastructure displayed increased neural vigilance, suggesting a lack of attention restoration, accompanied by reduced SART performance. These consistent findings offer empirical support for the Attention Restoration Theory, highlighting the effectiveness of tree exposure in enhancing attentional functioning. Future research should investigate the potential impact of bioswales on attention restoration.

Connections-between Daily Greenness Exposure and Health Outcomes.

A compelling body of research demonstrates that exposure to nature, especially trees, is beneficial to human health. We know little, however, about the extent to which understory vegetation that does not reach the height of trees, impacts human health. An additional gap in our knowledge concerns the extent to which daily variations in exposure to various forms of vegetation are related to human health outcomes. Many previous findings describing such connections were achieved in laboratory settings or through semi-controlled experiments, which do not reflect the dynamic variations of people's daily exposure to nature. Thus, we conducted an online survey to address these questions. We used the National Land Cover Dataset 2011 and Google Street View images to estimate participants' daily exposure to nature, and two standard questionnaires (General Health SF-12 and the Perceived Stress Scale) to assess health. Results show that greater exposure to trees in daily life is associated with better health outcomes. Specifically, higher neighborhood concentrations of tree canopy are related to better physical health, overall health and an increased capacity to control stress. In contrast, the results exploring the health associations of understory vegetation were inconsistent. In most cases, understory vegetation had a negative relationship with stress and mental health measures.

Synthesis of novel estrogen receptor antagonists using metal-catalyzed coupling reactions and characterization of their biological activity.

Estrogen receptor (ER) antagonists are valuable in the treatment of ER-positive human breast cancer. In this study, we designed and synthesized nine new derivatives of 17ß-estradiol (E2) with a bulky side chain attached to its C-7α position, and determined their ER antagonistic activity using in vitro bioassays. Four of the derivatives showed a strong inhibition of ERα transactivation activity in a luciferase reporter assay and blocked ERα interactions with coactivators. Similarly, these derivatives also strongly inhibited the growth of the ERα-positive human breast cancer cells. Computational docking analysis was conducted to model the interaction of these antagonists with the human ERα and showed that they could tightly bind to the ERα in a manner similar to that of ICI-182,780, a pure ER antagonist. These results provide an example that attachment of a bulky side chain to the C-7α position of E2 can produce ER antagonists with ER affinity comparable to that of ICI-182,780.

[Investigation on health workforce in county anti-schistosomiasis institutions of Jiangxi Province, 2010].

OBJECTIVE: To understand and assess changes of health workforce in county anti-schistosomiasis institutions of Jiangxi Province, so as to provide the evidence for improving the health workforce construction. METHODS: The status of health workforce of county anti-schistosomiasis institutions was surveyed with self-designed questionnaire, and the results were compared with the baseline survey results in 2007. RESULTS: The total number of employees was 1 384 in 2010 with the decrease of 71 persons compared with that in 2007. The average number of people per institution was 40.71, with the decrease of 0.86 per institution, compared with that of 2007. Compared with 2007, the employees with bachelor degree increased by 6.17%, those with college degree increased by 6.8%, and the attending rate of training and further study also increased. CONCLUSIONS: The health workforce has improved in county anti-schistosomiasis institutions of Jiangxi Province. The professional workers should be trained continually to improve their diathesis.

Chemical synthesis and biochemical characterization of a biotinylated derivative of 17beta-estradiol with a long side chain covalently attached to its C-7alpha position.

High-affinity biotinylated derivatives of 17beta-estradiol (E2) are of value for isolation of various estrogen-binding proteins (including estrogen receptors) and also for studying protein-protein interactions involving these proteins. In this study, we developed a simplified route for the chemical synthesis of a biotinylated derivative of E2 (compound 7) with a side chain attached to its C-7alpha position. Compound 7, i.e., 7alpha-{7-[8-(biotinamido)-octanamido]-heptyl}-estradiol, could be readily synthesized from 6-keto-estradiol-3,17beta-di-tetrahydropyranyl ether (compound 2, which can be prepared from E2), with a final yield of 36%. In vitro receptor-binding assay confirmed that the synthesized affinity ligand has a high binding affinity for both human estrogen receptor alpha and beta. When the affinity ligand (compound 7) was immobilized with avidin on an affinity column, it effectively bound human estrogen receptor alpha, and the receptor protein could be selectively eluted with a biotin-containing buffer. Using the same affinity ligand, prolyl 4-hydroxylase beta-subunit (also known as protein disulfide isomerase) was identified as one of the high-affinity E2-binding proteins in the whole cytosolic protein mixture prepared from MCF-7 human breast cancer cells. Computational molecular modeling analysis showed that compound 7 can bind to human estrogen receptor alpha in a similar manner as ICI-182,780 and raloxifene, and their binding energy values are also similar.

Chemical synthesis of six novel 17beta-estradiol and estrone dimers and study of their formation catalyzed by human cytochrome P450 isoforms.

Our earlier studies have shown that over 20 nonpolar 17beta-estradiol metabolite peaks were detected following incubations of radioactive 17beta-estradiol with human liver microsomes or recombinant human cytochrome P450 isoforms in the presence of NADPH as a cofactor. The structures of two representative nonpolar metabolites were identified earlier as dimers of 17beta-estradiol linked through a diaryl ether bond between the C-3 phenolic oxygen of one molecule and the C-2 or C-4 aromatic carbon of another. Six additional putative dimers between estrone and 17beta-estradiol with structures similar to the two identified ones were synthesized in this study. Using these newly synthesized estrogen dimers as reference standards, we demonstrated that incubations of human liver microsomes or various human cytochrome P450 isoforms with estrone or 17beta-estradiol alone or two estrogens in combination in the presence of NADPH as a cofactor resulted in the formation of all eight estrogen dimers in varying quantities.

Quantitative structure-activity relationship of various endogenous estrogen metabolites for human estrogen receptor alpha and beta subtypes: Insights into the structural determinants favoring a differential subtype binding.

To search for endogenous estrogens that may have preferential binding affinity for human estrogen receptor (ER) alpha or beta subtype and also to gain insights into the structural determinants favoring differential subtype binding, we studied the binding affinities of 74 natural or synthetic estrogens, including more than 50 steroidal analogs of estradiol-17beta (E2) and estrone (E1) for human ER alpha and ER beta. Many of the endogenous estrogen metabolites retained varying degrees of similar binding affinity for ER alpha and ER beta, but some of them retained differential binding affinity for the two subtypes. For instance, several of the D-ring metabolites, such as 16 alpha-hydroxyestradiol (estriol), 16 beta-hydroxyestradiol-17 alpha, and 16-ketoestrone, had distinct preferential binding affinity for human ER beta over ER alpha (difference up to 18-fold). Notably, although E2 has nearly the highest and equal binding affinity for ER alpha and ER beta, E1 and 2-hydroxyestrone (two quantitatively predominant endogenous estrogens in nonpregnant woman) have preferential binding affinity for ER alpha over ER beta, whereas 16 alpha-hydroxyestradiol (estriol) and other D-ring metabolites (quantitatively predominant endogenous estrogens formed during pregnancy) have preferential binding affinity for ER beta over ER alpha. Hence, facile metabolic conversion of parent hormone E2 to various metabolites under different physiological conditions may serve unique functions by providing differential activation of the ER alpha or ER beta signaling system. Lastly, our computational three-dimensional quantitative structure-activity relationship/comparative molecular field analysis of 47 steroidal estrogen analogs for human ER alpha and ER beta yielded useful information on the structural features that determine the preferential activation of the ER alpha and ER beta subtypes, which may aid in the rational design of selective ligands for each human ER subtype.

Synthesis of 7alpha-substituted derivatives of 17beta-estradiol.

Estrogen receptor (ER) pure antagonists such as ICI-182,780 (fulvestrant) are effective alternatives to tamoxifen (an ER antagonist/weak partial agonist) in the treatment of postmenopausal, receptor-positive human breast cancers. Structurally, these pure antagonists contain the basic core structure of 17beta-estradiol (E(2)) with a long side chain attached to its C-7alpha position. We explored and compared in this study various synthetic routes for preparing a number of C-7alpha-substituted derivatives of E(2), which are highly useful for the design and synthesis of high-affinity ER antagonists, ER-based imaging ligands, and other ER-based multi-functional agents. Using E(2) as the starting material and 1-iodo-6-benzyloxyhexane as a precursor for the C-7alpha side chain, a seven-step synthetic procedure afforded 3,17beta-bis(acetoxy)-7alpha-(6-hydroxyhexanyl)-estra-1,3,5(10)-triene (one of the derivatives prepared) in an overall yield of approximately 45% as compared to other known procedures that afforded substantially lower overall yield (8-27%). The synthetic steps for this representative compound include: (1) protection of the C-3 and C-17beta hydroxyls of E(2) using methoxymethyl groups; (2) hydroxylation of the C-6 position of the bismethoxymethyl ether of E(2); (3) Swern oxidation of the C-6 hydroxy to the ketone group; (4) C-7alpha alkylation of the C-6 ketone derivative of E(2); (5) deprotection of the two methoxymethyl groups; (6) reprotection of the C-3 and C-6 free hydroxyls with acetyl groups; (7) removal of the C-6 ketone and the benzyl group on the side chain by catalytic hydrogenation in acetic acid. As predicted, two of the representative C-7alpha-substituted derivatives of E(2) synthesized in the present study retained strong binding affinities (close to those of E(2) and ICI-182,780) for the human ERalpha and ERbeta subtypes as determined using the radioligand-receptor binding assays.

Reactions of the putative neurotoxin tryptamine-4,5-dione with L-cysteine and other thiols.

Tryptamine-4,5-dione (1) is formed by oxidation of 5-hydroxytryptamine by reactive oxygen and reactive nitrogen species. Dione 1 is a powerful electrophile that can covalently modify cysteinyl residues of proteins and deactivate key enzymes. Thus, 1 has been suggested to play a role in the degeneration of serotonergic neurons in brain disorders such as Alzheimer's disease or evoked by amphetamine drugs. However, if formed in the brain, it is also likely that 1 would react with low molecular weight thiols such as cysteine (CySH) and glutathione (GSH). The resulting metabolites might not only contribute to the degeneration of serotonergic neurons but also, perhaps, serve as biomarkers of such neurodegeneration. In this investigation, it is shown that in oxygenated buffer at pH 7.4 dione 1 reacts with CySH and other low molecular weight sulfhydryls such as GSH, N-acetylcysteine, and cysteamine to form, first, the corresponding 7-S-thioethers of the dione. However, unlike the glutathionyl and N-acetylcysteinyl conjugates of 1, the 7-S-cysteinyl conjugate is very unstable at pH 7.4 forming a number of novel products, the nature of which are dependent on the relative concentrations of 1 and CySH. These products have been isolated, and spectroscopic and other evidence is provided in support of their proposed chemical structures.

Stability of the putative neurotoxin tryptamine-4,5-dione.

Tryptamine-4,5-dione (1) is formed by oxidation of the neurotransmitter 5-hydroxytryptamine by reactive oxygen and reactive nitrogen species, and on the basis of in vitro and in vivo studies, it has been proposed to be a neurotoxin that may contribute to the selective neurodegeneration in Alzheimer's disease and the serotonergic neurotoxicity of methamphetamine. Several investigators have noted that under the conditions employed in the past to synthesize 1 and explore its in vitro and in vivo biological properties, the dione is somewhat unstable. In the present study, the stability of 1 has been investigated in a number of media employed in previous investigations to synthesize the dione and evaluate its biological properties. At low concentrations (< or =200 micro M), 1 is most stable in artificial cerebrospinal fluid (aCSF, pH 6-6.5) in which it decomposes < or =10% over 24 h forming primarily 3-(2-aminoethyl)-6-[3'-(2-aminoethyl)-indol-4',5'-dione-7'-yl]-5-hydroxyindole-4,7-dione (10). In phosphate buffer or 0.5 M NH(4)Cl solutions at pH 7.4 and in acidic solution (e.g., 0.01 M HCl), such low concentrations of 1 also decompose to 10 although somewhat more rapidly than in aCSF. As the concentration of 1 is increased in all of these media, its decomposition becomes more rapid and shifts toward formation of 7,7'-bi-(5-hydroxytryptamine-4-one) (9) and its autoxidation product 7,7'-bitryptamine-4,5-dione (11). At 20 mM concentrations in aCSF or at pH 7.4, 1 rapidly decomposes to a dark, uncharacterized, presumably polymeric precipitate. However, in 0.01 M HCl solution >/=20 mM, 1 rapidly and almost quantitatively dimerizes to 9. The initial reaction of 1, which leads to the ultimate formation of 9 or 11 and 10, is the nucleophilic addition of water to the C(7) position of the dione to form 4,5,7-trihydroxytryptamine (2). Oxidation of 2 by 1 and/or molecular oxygen forms radical species, the predominant form of which has been detected by electron spin resonance spectroscopy using a spin stabilization method. Subsequent reactions of radical intermediates lead to the formation of 9 or 11 and 10. The results of this investigation are discussed in terms of previous in vitro and in vivo biological properties of 1 and its possible role in the serotonergic neurotoxicity of methamphetamine and neurodegenerative diseases.

Structural characterization of a case-implicated contaminant, "Peak X," in commercial preparations of 5-hydroxytryptophan.

OBJECTIVE: To determine the chemical structure of a contaminant, X1, previously found in eosinophilia myalgia syndrome case-implicated 5-hydroxytryptophan (5-OHTrp), and also present in over-the-counter (OTC) commercially available 5-OHTrp. METHODS: Case-implicated 5-OHTrp as well as 6 OTC samples were subjected to accurate mass HPLC-mass spectrometry and HPLC-electrochemical detection, and reacted with reduced glutathione. Peak X1 was subsequently subjected to HPLC-tandem mass spectrometry (MS/MS), as well as the resulting nucleophilic glutathione product. All these data were compared with analysis carried out under identical conditions on authentic 4,5-tryptophan-dione (Trp-4,5D). RESULTS: Based on accurate mass, tandem mass spectrometric analysis, and comparision with authentic standard compound analysis, X1 was determined to be 4,5-tryptophan-dione, a putative neurotoxin. The presence of X1 in OTC samples varied from 0.5 to 10.3% of the amount of Trp-4,5D present in case-implicated 5-OHTrp. CONCLUSION: Peak X1 was identified as the putative neurotoxin Trp-4,5D. It was found in case-implicated 5-OHTrp as well as 6 OTC samples. This gives some cause for concern in terms of the safety of such commercial preparations of 5-OHTrp.

Inhibition of the alpha-ketoglutarate dehydrogenase and pyruvate dehydrogenase complexes by a putative aberrant metabolite of serotonin, tryptamine-4,5-dione.

A transient energy impairment with resultant release and subsequent reuptake of 5-hydroxytryptamine (5-HT) and NMDA receptor activation with consequent cytoplasmic superoxide (O(2)(-)(*)), nitric oxide (NO(*)), and peroxynitrite (ONOO(-)) generation have all been implicated in a neurotoxic cascade which ultimately leads to the degeneration of serotonergic neurons evoked by methamphetamine (MA) and 3,4-methylenedioxymethamphetamine (MDMA). Such observations raise the possibility that the O(2)(-)(*)/NO(*)/ONOO(-)-mediated oxidation of 5-HT, as it returns via the plasma membrane transporter to the cytoplasm of serotonergic neurons when the MA/MDMA-induced energy impairment begins to subside, may generate an endogenous neurotoxin. In vitro the O(2)(-)(*)/NO(*)/ONOO(-)-mediated oxidation of 5-HT forms tryptamine-4,5-dione (T-4,5-D). When incubated with intact rat brain mitochondria, T-4,5-D strongly inhibits state 3 respiration with pyruvate or alpha-ketoglutarate as substrates at concentrations which do not affect succinate-supported (complex II) respiration. Experiments with freeze-thawed rat brain mitochondria reveal that T-4,5-D inhibits the pyruvate dehydrogenase and alpha-ketoglutarate dehydrogenase complexes. These and other properties of T-4,5-D raise the possibility that it may be an endogenously formed intraneuronal metabolite of 5-HT that contributes to the serotonergic neurotoxicity of MA and MDMA.

Seek protein which can interact with hepatitis B virus X protein from human liver cDNA library by yeast two-hybrid system.

AIM: To seek the X associated protein (XAP) with the constructed bait vector pAS2-1X from normal human liver cDNA library. METHODS: The X region of the HBV gene was amplied by PCR and cloned into the eukaryotic expression vector pAS2-1. The reconstituted plasmid pAS2-1X was transformed into the yeast cells and the expression of X protein (pX) was confirmed by Western blot analysis. Yeast cells were cotransformed with pAS2-1X and the normal human liver cDNA library and were grown in selective SC/-trp-leu-his-ade medium, the second screen was performed with the LacZ report gene. Furthermore, segregation analysis and mating experiment were performed to eliminate the false positive and the true positive clones were selected for PCR and sequencing. RESULTS: Reconstituted plasmid pAS2-1X including the anticipated fragment of X gene was proved by auto-sequencing assay. Western blot analysis showed that reconstituted plasmid pAS2-1X expressed BD:X fusion protein in yeast cells. Of 5 x 10(6) transformed colonies screened,65 grew in the selective SC/-trp-leu-his-ade medium, 5 scored positive for beta-gal activity, and only 2 remaining clones passed through the segregation analysis and mating experiment. Sequence analysis identified that two clones contained similar cDNA fragment:GAACTTGCG. CONCLUSION: The short peptide(glutacid-leucine-alanine)is a possible required site for XAP binding to pX. Normal human liver cDNA library has difficulties in expressing the integrated XAP on yeast cells.