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Children are more likely to experience maltreatment and parental conflict in a pandemic context, which can exacerbate their vulnerability to psychological disorders. The purpose of the present study was to examine mental health symptoms in children aged 0 to 10 years and consider related factors from the perspectives of maltreatment and parental conflict during the COVID-19 lockdown. Participants were 1286 parents aged 18 years and over with children aged 0 to 10 years were included. Several multivariable linear regressions were used to analyze the data. The largest variance in child mental health was explained by child maltreatment, as more maltreatment predicted higher reported psychological problems (standardized beta = 0.49, P 0.001). Comparatively, parental conflict predicted less variance in mental health problems than maltreatment (standardized beta = 0.18, P 0.001). Children who experienced more maltreatment experience and exposure to COVID-19 showed elevated levels of mental health symptoms (standardized beta = 0.06, p 0.05), as did those who experienced parental conflict and pandemic exposure (standardized beta = 0.06, p 0.05). The findings highlight that tailored programs that focus on a healthy family environment and strategic parental support services may be particularly effective in reducing children's mental health problems due to COVID-19 exposure.

Background Previous studies have suggested that sensory loss is linked to depression. However, most of these studies have been conducted in developed countries and the results are mixed. Objectives The current study aims to examine the longitudinal relationship between hearing loss, vision loss, dual sensory loss, and depression among Chinese older adults over four years. Methods The data were obtained from the China Health and Retirement Longitudinal Survey (CHARLS). In total, 6353 Chinese older adults aged 60 years and over were included at baseline in this study. Self-reported hearing and vision status was used, and dual sensory loss was a combined variable of hearing loss and vision loss. Depressive symptoms were assessed by The Center for Epidemiologic Studies Depression Scale (CESD-10). The baseline relationship between sensory loss and depression was explored by logistic regression analyses. A logistic mixed model was used to assess whether baseline sensory loss was associated with incident depression for older adults with no depressive symptoms at baseline over four years. Results At baseline, vision loss (OR=1.46, 95%CI: 1.19-1.79) and dual sensory loss (OR=2.14, 95%CI: 1.77-2.58) were associated with depression, while hearing loss was not. After four years, vision loss (OR=1.60, 95%CI: 1.16-2.22) and dual sensory loss (OR=1.78, 95%CI: 1.32-2.41) were associated with incident depression. Limitations Not all potential confounding factors are measured and adjusted in the analysis. Apart from that, all measurements of interest are self-reported scales. Conclusion Vision loss and dual sensory loss are significantly associated with both onset and increased depressive symptoms over time, while hearing loss is not associated with depression. Interactions between all types of sensory loss and social activities are not significant. Our results suggest that target mental health intervention programs should be delivered to vision loss or dual sensory loss populations.

In the field of environmental science and engineering, microorganisms, enzymes and algae are promising biomass materials that can effectively degrade pollutants. However, problems such as poor environmental adaptability, recycling difficulties, and secondary pollution exist in the practical application of non-immobilized biomass materials. Biomass immobilization is a novel environmental remediation technology that can effectively solve these problems. Compared with non-immobilized biomass, immobilized biomass materials have the advantages of reusability and stability in terms of pH, temperature, handling, and storage. Many researchers have studied immobilization technology (i.e., methods, carriers, and biomass types) and its applications for removing refractory organic pollutants. Based on this, this paper reviews biomass immobilization technology, outlines the mechanisms and factors affecting the removal of refractory organic pollutants, and introduces the application of immobilized biomass materials as fillers for reactors in water purification. This review provides some practical references for the preparation and application of immobilized biomass materials and promotes further research and development to expand the application range of this material for water purification.

Supplemental blue/red lighting accelerated fruit coloring and promoted lycopene synthesis in tomato fruits. Potassium (K) is the most enriched cation in tomato fruits, and its fertigation improved tomato yield and fruit color. However, the effects of supplemental lighting on K uptake and transport by tomatoes and whether supplemental lighting accelerates fruit coloring through enhancing K uptake and transport are still unclear. We investigated the effects of supplemental light-emitting diode (LED) lighting (SL, 100% red, 100% blue, 75% red combined 25% blue) on K uptake in roots and transport in the fruits as well as the fruit coloring of tomatoes (Micro-Tom) grown in an experimental greenhouse in hydroponics. The use of red SL or red combined blue SL enhanced K uptake and K accumulation as well as carotenoid (phytoene, lycopene, γ-carotene, and β-carotene) content in fruits by increasing photosynthesis, plant growth, and fruit weight. The genes related to ethylene signaling were upregulated by red SL. Quantitative real-time PCR (qRT-PCR) results showed that K transporter genes (SlHAKs) are differentially expressed during fruit development and ripening. The highest-expressed gene was SlHAK10 when fruit reached breaker and ripening. SlHAK3 and SlHAK19 were highly expressed at breaker, and SlHAK18 was highly expressed at ripening. These might be related to the formation of tomato fruit ripening and quality. SlHAK4, SlHAK6, SlHAK8, and SlHAK9 were significantly downregulated with fruit ripening and induced by low K. The expression level of SlHAK6, SlHAK10, SlHAK15, and SlHAK19 were significantly increased by blue SL or red combined blue SL during breaker and ripening. Blue SL or red combined blue SL increased content of phytoene, β-carotene, α-carotene, and γ-carotene and accelerated fruit coloring by enhancing K uptake in roots and transport in fruits during fruit ripening. This was consistent with the expression level of SlHAK6, SlHAK10, SlHAK15, and SlHAK19 during fruit development and ripening. The key genes of photoreceptors, light signaling transcript factors as well as abscisic acid (ABA) transduction induced by blue SL or red combined blue SL were consistent with the upregulated genes of SlHAK6, SlHAK10, SlHAK15, and SlHAK19 under blue SL and red combined blue SL. The K transport in tomato fruits might be mediated by light signaling and ABA signaling transduction. These results provide valuable information for fruit quality control and the light regulating mechanism of K transport and fruit coloring in tomatoes.

In this study, a high-performance adsorbent Co@AC was prepared by loading cobalt ions (Co2+) on activated carbon (AC) via solution impregnation and high-temperature calcination technology, and was used to remove atrazine in water. The preparation factors on the adsorbent properties were studied, and the characteristics were analyzed by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), and Fourier transform infrared spectrometer (FTIR). The results showed that Co@AC possessed the best performance when the factors were 7.0% of Co2+ (w/v), 7.0 h of immersing time, 500 &deg, C of calcination temperature and 4.0 h of calcination time. The adsorption conditions and mechanisms for atrazine removal by Co@AC were also studied scientifically. As the conditions were pH 4.0, reaction time 90 min and temperature 25 &deg, C, Co@AC had the largest adsorption capacity, which was 92.95 mg/g, and the maximum removal rate reached 94.79%. The correlation coefficient of the Freundlich isotherm was better than that of the Langmuir isotherm, and the adsorption process followed the pseudo-second-order kinetic model. Cycle experiments showed that the removal efficiency of atrazine by Co@AC remained above 85% after five repeated experiments, indicating that Co@AC showed a strong stable performance and is a promising material for pesticides removal.

Background Detection of depression gained prominence soon after this troublesome disease emerged as a serious public health concern worldwide. Objective This systematic review aims to summarize the findings of previous studies concerning applying machine learning (ML) methods to text data from social media to detect depressive symptoms and to suggest directions for future research in this area. Methods A bibliographic search was conducted for the period of January 1990 to December 2020 in Google Scholar, PubMed, Medline, ERIC, PsycINFO, and BioMed. Two reviewers retrieved and independently assessed the 418 studies consisting of 322 articles identified through database searching and 96 articles identified through other sources; 17 of the studies met the criteria for inclusion. Results Of the 17 studies, 10 had identified depression based on researcher-inferred mental status, 5 had identified it based on users’ own descriptions of their mental status, and 2 were identified based on community membership. The ML approaches of 13 of the 17 studies were supervised learning approaches, while 3 used unsupervised learning approaches; the remaining 1 study did not describe its ML approach. Challenges in areas such as sampling, optimization of approaches to prediction and their features, generalizability, privacy, and other ethical issues call for further research. Conclusions ML approaches applied to text data from users on social media can work effectively in depression detection and could serve as complementary tools in public mental health practice.

BACKGROUND Detection of depression gained prominence soon after this troublesome disease emerged as a serious public health concern worldwide. OBJECTIVE This systematic review aims to summarize the findings of previous studies concerning applying machine learning (ML) methods to text data from social media to detect depressive symptoms and to suggest directions for future research in this area. METHODS A bibliographic search was conducted for the period of January 1990 to December 2020 in Google Scholar, PubMed, Medline, ERIC, PsycINFO, and BioMed. Two reviewers retrieved and independently assessed the 418 studies consisting of 322 articles identified through database searching and 96 articles identified through other sources; 17 of the studies met the criteria for inclusion. RESULTS Of the 17 studies, 10 had identified depression based on researcher-inferred mental status, 5 had identified it based on users’ own descriptions of their mental status, and 2 were identified based on community membership. The ML approaches of 13 of the 17 studies were supervised learning approaches, while 3 used unsupervised learning approaches; the remaining 1 study did not describe its ML approach. Challenges in areas such as sampling, optimization of approaches to prediction and their features, generalizability, privacy, and other ethical issues call for further research. CONCLUSIONS ML approaches applied to text data from users on social media can work effectively in depression detection and could serve as complementary tools in public mental health practice.

Highlights • A larger number of ACEs predict substantial higher levels of PTSS and anxiety. • Experienced exposure and fear of exposure also predict more PTSS and anxiety. • Childhood abuse and neglect may exacerbate the impact of exposure on mental health., Background Since the COVID-19 outbreak at the end of 2019, it has evolved into a global pandemic with tremendous mental health impact besides the threats to people’s physical health. Objective The aims were to examine whether exposure to COVID-19 predicts elevated levels of anxiety and post-traumatic stress symptoms and whether pre-pandemic maltreatment experiences exacerbate this impact on mental health in adolescents. Participants and setting The survey was conducted online from February 8 st to February 27th, 2020, and the questionnaires were distributed and retrieved through a web-based platform. This study includes a total of 6196 subjects, aged range from 11 to 18 years old. Methods Several multivariable linear regressions were used to analyse the data. Results The largest variance in PTSS and anxiety problems was explained by ACEs, with more pre-pandemic maltreatment experiences predicting more PTSS (effect size beta = 0.16∼0.27), and more anxiety (effect size beta = 0.32∼0.47). Experienced or subjective fear of exposure to COVID-19 predicted statistically significant variance in PTSS and anxiety, and standardized betas ranged from 0.04 to 0.09. Participants who had adverse childhood experiences and had experienced exposure to COVID-19 showed elevated PTSS. Conclusions After pre-pandemic maltreatment experiences the impact of exposure to COVID-19 on mental health may be stronger. Scars from the past seem to be vulnerabilities during societal upheaval. We therefore suggest that when exposed to COVID-19 rural adolescents should get prioritized professional family support and mental health counseling in particular when they have experienced family abuse and neglect in childhood, even though such support is more difficult to organize in rural areas.

Internet addiction among adolescents is an emerging public health issue. The aim of this study was to examine the relationship between being bullied and internet addiction, and further to test the mediating effect of adult attachments on this relationship among Chinese rural adolescents. A total of 1,270 adolescents from three high schools in Henan province were recruited as subjects. Internet addiction was measured by Young’s internet addiction scale. This study showed that the prevalence of being bullied and addictive internet use among the recruited Chinese rural adolescents were 41.97% and 11.34%, respectively. Being bullied was significantly associated with internet addition. Adult attachment was found to mediate the association between being bullied and internet addiction. Higher anxiety attachment and lower close-dependent attachment partially explained the higher risk of internet addiction among the Chinese rural adolescents being bullied. This study suggests that the experience of being bullied can increase the risk of addictive internet use and that this risk can be mediated through adult attachments. This discovery supports the adoption of effective prevention programs to decrease school bullying and to improve adult attachments for adolescents in China and other countries.

The mental health problems might have been increased owing to the COVID-19 pandemic with the commencement of the year 2020, therefore, an epidemiological survey appraising the burden of mental health issues among the general population is imperative. This cross-sectional study attempts to reveal the underlying mental health conditions, such as Post-Traumatic Stress Symptoms (PTSS), depression, and insomnia, relating to the pandemic situation, and to further examine the combined effects of gender and age on the COVID-19 related mental health consequences. An online survey was conducted among 2,992 adults in China from February 1st 2020 to February 10th 2020. The study uses binary logistic regression to analyze the potential factors associated with PTSD, depression, and insomnia. The results indicate that the prevalence of PTSS, depression, and insomnia are 19.5, 26.9, and 19.6% respectively during the COVID-19. Men and women show different rates of PTSS and depression, whereas no insomnia is found in both males and females. The females above 50 years of age have a lower level of depressive symptoms (OR = 0.448, 95%CI: 0.220–0.911, Cohen's d = −0.443) as compared with females aged 18–25; while the highest effect sizes for PTSS (OR = 2.846, 95%CI: 1.725–4.695, Cohen's d = 0.537) and the depression (OR = 2.024, 95%CI: 1.317–3.111, Cohen's d = 0.314) are seen in males aged 26 to 30. Besides gender, education, living conditions, direct exposure to COVID-19, the post mental and the physical health condition is related to PTSS, depression, and insomnia. Our study suggests that high-risk groups, especially those having two or more related factors and young men, should be the focus of mental health intervention.

The specific impacts of attitudes toward aging on depressive symptoms have not been widely reported in previous studies in China.The aim is to examine the associations between attitudes toward aging, perceived social support, and depressive symptoms among older adults stratified by rural and urban dwelling.This study used a cross-sectional data including 7209 participants, among which 64.6% were urban adults and 35.4% were rural adults. Several multiple liner regression models were used to analysis the data. Three social support types were analyzed as moderators of the relationship between the attitudes toward aging and depressive symptoms.Positive attitudes toward aging (β=-0.139, P0.001), negative attitudes toward aging (β=0.284, P0.001) were significantly associated with lower depressive symptoms among older Chinese adults. Support from family (β=-0.087, P0.001), friends (β=-0.047, P0.01) and the government (β=-0.035, P0.01) were all significantly associated with urban older adults' levels of depressive symptoms. Only family support (β=-0.109, P0.001) was associated with lower depressive symptoms among rural older adults'. In addition, family support buffered the effect of negative attitudes toward aging on depressive symptoms for all the older adults, while the moderation effects of support from friends and government only worked for urban elderly.A cross-sectional design is limited to establish causal associations.Addressing depression among older adults should focus on improving attitudes toward aging and expanding the availability of social support. Moreover, deeper reforms are needed to address inequalities between urban and rural areas in China.

The kynurenine (Kyn) pathway is the major route for tryptophan (Trp) metabolism, and it contributes to several fundamental biological processes. Trp is constitutively oxidized by tryptophan 2, 3-dioxygenase in liver cells. In other cell types, it is catalyzed by an alternative inducible indoleamine-pyrrole 2, 3-dioxygenase (IDO) under certain pathophysiological conditions, which consequently increases the formation of Kyn metabolites. IDO is up-regulated in response to inflammatory conditions as a novel marker of immune activation in early atherosclerosis. Besides, IDO and the IDO-related pathway are important mediators of the immunoinflammatory responses in advanced atherosclerosis. In particular, Kyn, 3-hydroxykynurenine, and quinolinic acid are positively associated with inflammation, oxidative stress (SOX), endothelial dysfunction, and carotid artery intima-media thickness values in end-stage renal disease patients. Moreover, IDO is a potential novel contributor to vessel relaxation and metabolism in systemic infections, which is also activated in acute severe heart attacks. The Kyn pathway plays a key role in the increased prevalence of cardiovascular disease by regulating inflammation, SOX, and immune activation.

This study explores the ability of a catalytic antioxidant, Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP), to protect against neuronal and glial oxidative stress and death after spinal cord injury (SCI). Nine different doses of MnTBAP were administered into the intrathecal space of the rat spinal cord immediately following moderate SCI to establish dose - response curves for prevention of lipid peroxidation and neuron death. An optimal dose was determined by comparing the effectiveness of MnTBAP protection among doses. The optimal dose was then administered and the cords were removed 24 h post-administration and processed for staining. The cells in the cord sections at different distances from the epicenter were counted to obtain the spatial profiles of MnTBAP protection. Comparison of the counts between MnTBAP- and vehicle-treated groups in the sections double immuno-fluorescence-stained with oxidative and cellular markers demonstrated that MnTBAP significantly reduced numbers of nitrotyrosine- and DNP-positive (stained with an antibody against 2,4-dinitrophenyl hydrazine (DNPH)-labeled protein carbonyls) neurons, astrocytes, and oligodendrocytes. Comparison of the counts between the two treatments in the sections immuno-stained with cellular markers revealed that MnTBAP significantly increased numbers of neurons, motoneurons, astrocytes, and oligodendrocytes. MnTBAP more effectively reduced neuronal than glial cell death. Post-injury treatment with the optimal dose of MnTBAP at 6, 12, 24, 48, and 72 h post-SCI demonstrated that the effective time window for reducing protein nitration and neuron death was at least 12 h. Our results demonstrated that MnTBAP combats oxidative stress, thereby attenuating all types of cell death after SCI.

This study measured the time courses of concentration changes following administration of the catalytic antioxidants Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) and Mn (III) 3-methoxy N, N' bis (salicyclidene) ethylenediamine chloride (EUK-134) in blood and cerebrospinal fluid (CSF) of rats with a spinal cord injury (SCI) and sham controls. Parallel measurements were made for methylprednisolone, the only drug presently used clinically for treating SCI. The time courses kinetically characterized the agents in their stability, disposition, and ability to penetrate the blood–spinal cord barrier (BSB). In both the SCI and control groups, MnTBAP was stable in CSF and in blood across the collection periods (10 h and 24 h, respectively) following administration. In the blood, [EUK-134] and [methylprednisolone] rapidly declined to near basal concentrations at 4 h and 2 h, respectively, post-administration. Therefore the order of stability in CSF and blood was MnTBAP >> EUK-134 > methylprednisolone. The maximum CSF/blood concentration ratios for EUK-134, methylprednisolone and MnTBAP post-administration were: 32 ± 3.1%, 19.2 ± 6.4%, and 4.42 ± 0.73% in the injured rats, and 22 ± 6.5%, 17.8 ± 2.9%, and 1.0 ± 0.5% in the sham control animals. This suggests an order of BSB penetration of EUK-134 > methylprednisolone >> MnTBAP. Despite much lower penetration by MnTBAP compared with EUK-134 and methylprednisolone, a lower dose of MnTBAP because of its stability provided a higher concentration in CSF than did the other agents given at higher doses. This finding supports further exploration of MnTBAP as a potential treatment for SCI.

The purpose of this study was to investigate the changes in the gene expression of Mitofusion (Mfn) 1 and 2 and Fission 1 (Fis1) and mitochondrial energy metabolism in response to altered energy demand during prolonged exercise in rat skeletal muscle. Male Sprague-Dawley rats were subjected to an acute bout of treadmill running at various durations and killed immediately or during recovery. Mfn1/2 and Fis1 mRNA and protein contents, reactive oxygen species (ROS) generation, state 3 and state 4 respiration rates, trans-innermembrane potential and ATP synthase activity were measured in isolated muscle mitochondria. We found that (1) Mfn1/2 mRNA contents were progressively decreased during 150 min of exercise, along with decreased Mfn 1 protein levels. Fis1 mRNA and protein contents showed significant increases after 120-150 min of exercise. These changes persisted through the recovery period up to 24 h. (2) Mitochondrial ROS generation and state 4 respiration showed progressive increases up to 120 min, but dropped at 150 min of exercise. (3) State 3 respiration rate and respiratory control index were unchanged initially but decreased at 150 and 120 min of exercise, respectively, whereas ATP synthase activity was elevated at 45 min and returned to resting level thereafter. Our data suggested that the gene expression of mitochondrial fusion and fission proteins in skeletal muscle can respond rapidly to increased metabolic demand during prolonged exercise, which could significantly affect the efficiency of oxidative phosphorylation.

This study presents quantitative temporal and spatial profiles of neuronal loss and apoptosis following a contusion spinal cord injury (50 g · cm). The profiles were evaluated by counting the cresol violet–stained surviving cells and the total number of TUNEL-positive cells and of TUNEL-positive neurons in sections 0– 4 mm from the epicenter and 1, 6, 12, 24, 48, and 72 hr and 1 week postinjury. We demonstrated that neurons continue to disappear over 1 week postinjury and that neuronal loss shifts to areas longer distances from the epicenter over time. TUNEL-positive cells in both gray and white matter appeared after 6 hr, gradually increased to a peak level after 48 hr, and declined by 72 hr postinjury. TUNEL-positive neurons peaked earlier and were present for 1 week, although the total number of neurons was reduced significantly by the end of the week. The neuronal loss and apoptosis were partially prevented by a metalloporphyrin [Mn(III) tetrakis (4-benzoic acid) porphyrin (MnTBAP)]. We demonstrated that MnTBAP (10 and 50 mg/kg, given intraperitoneally) significantly reduced neuronal death in the sections 1–2.5 mm rostral and 1 mm caudal from the epicenter compared with that in the vehicle-treated group, suggesting MnTBAP is more effective in the sections rostral than in those caudal to the epicenter. MnTBAP (10 mg/kg) significantly reduced the number of TUNEL-positive neurons in the sections 1 mm caudal from the epicenter. Our profiles provide a database for pharmacological intervention, and our results on MnTBAP treatment support an important role for antioxidant therapy in spinal cord injury. © 2007 Wiley-Liss, Inc.

The goal of the present study was to determine in vivo whether peroxynitrite, at the concentration and duration produced by SCI, contributes to membrane lipid peroxidation (MLP) after traumatic spinal cord injury (SCI) and the capability of a broad spectrum scavenger of reactive species, Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP), to reduce MLP. This was accomplished by administering a peroxynitrite donor 3-morpholinosydnonimine (SIN-1) into the gray matter of an uninjured rat spinal cord through a microdialysis fiber to generate ONOO at the SCI-elevated levels. The resulting MLP was characterized by measuring the productions of extracellular malondialdehyde and of intracellular 4-hydroxynonenal. We demonstrated that extracellular SIN- 1 administration significantly increased the concentration of malondialdehyde (p0.001) and the numbers of hydroxynonenal-positive cells (p0.001) as compared to a control group in which ACSF was administered. Simultaneous administration of MnTBAP through a second microdialysis fiber significantly reduced SIN-1-induced malondialdehyde production (p0.001) and the numbers of HNE-positive cells (p0.001). There was no significant difference between MnTBAP-treated and ACSF-controls (p = 0.3). These results demonstrate in vivo that (1) SCI-produced levels of peroxynitrite sufficient to cause MLP, and therefore that peroxynitrite is an agent of secondary damage after acute SCI; (2) MnTBAP can efficiently reduce SIN-1-induced MLP.

This study explores whether the hydroxyl radical (*OH)-one of the most destructive reactive oxygen species-plays a role in secondary spinal cord injury (SCI). First, we measured the time course of *OH formation in rat spinal tissue after impact SCI by administering salicylate as a trapping agent into the intrathecal space of the cord and measuring the hydroxylation products of salicylate, 2,3- and 2,5-dihydroxybenzoic acid (2,3- and 2,5-DHBA) by HPLC. The 2,3-DHBA concentration was significantly higher in injured spinal tissue than in sham controls at 5 min, 1 and 3 h, but not at 5 h post-injury. Second, we generated *OH by administering H(2)O(2) and FeCl(2)/EDTA (Fenton's reagents) at the concentrations produced by SCI into the gray matter of the cord for 4 h and found that it induced significant cell loss at 24 h post-*OH exposure. Mn (III) tetrakis (4-benzoic acid) porphyrin(MnTBAP)-a broad spectrum reactive species scavenger-significantly reduced *OH-induced cell death. Finally, we generated superoxide and administered FeCl(3)/EDTA in the intrathecal space of the cord at the concentration produced by SCI and measured extracellular *OH formation in the gray matter of the cord by microdialysis sampling. We found that the levels of *OH significantly increased compared to the pre-administration level, indicating that *OH can be produced in vivo by the iron-catalyzed Haber-Weiss reaction. All together, we demonstrated that *OH is an endogenous secondary damaging agent following SCI and the metal-catalyzed Haber-Weiss reaction may contribute to early *OH formation after SCI.

We previously measured the time courses of hydrogen peroxide (H2O2), hydroxyl radical (*OH), and catalytic iron increases following traumatic spinal cord injury (SCI). This study determines whether the SCI-elevated level of *OH causes cell death. OH was generated by administering H2O2 and Fe2+ at the concentrations attained following SCI, each through a separate microdialysis fiber inserted laterally into the gray matter of the cord. The duration of *OH generation mimics the duration of its elevation after SCI. The death of neurons and astrocytes was characterized at 24 h post-*OH exposure and quantitated by counting surviving cells along the fiber track in sections stained with Cresyl Violet, or immunohistochemically stained with anti-neuron-specific enolase (anti-NSE) and anti-glial fibrillary acidic protein (anti-GFAP). DNA fragmentation in neurons was characterized by double staining with terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) and anti-NSE. Using a one way ANOVA followed by the Tukey test, we demonstrated that *OH generated in the cord induced significant losses of neurons in both Cresyl Violet (P

This study used a weight drop impact injury model to explore the role of iron and the reality of iron-catalyzed hydroxyl radical ((*)OH) formation in secondary spinal cord injury (SCI). The time course of total extracellular iron was measured following SCI by microcannula sampling and atomic absorption spectrophotometry analysis. Immediately following SCI, the total iron concentration increased from an undetectable level to an average of 1.32 microM. The time course of SCI-induced (*)OH-generating catalytic activity in the cord was obtained by determining the ability of tissue homogenate to convert hydrogen peroxide to (*)OH and then measuring 2,3-dihydroxybenzoic acid, a hydroxylation product of salicylate. The concentration of 2,3-DHBA quickly and significantly increased (p

We previously demonstrated that the peroxynitrite concentration increases after impact spinal cord injury. This study tests whether spinal cord injury-elevated peroxynitrite induces apoptotic cell death. Peroxynitrite was generated at the concentration and duration produced by spinal cord injury by administering S-morpholinosydnonimine through a microdialysis fiber into the gray matter of the rat spinal cord. Fragmented DNA was visualized by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling. Transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling-positive neurons were quantitated by counting the transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling and neuron-specific enolase double-stained neurons along the fiber track in the sections removed at 6, 12, 24 and 48 h post-peroxynitrite exposure. Peroxynitrite significantly increased transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling-positive neurons at all time points examined (P≤0.001) compared with artificial cerebrospinal fluid controls (Two-way analysis of variance followed by Tukey test), peaking at 24 h post-exposure. Electron microscopic observation of characteristic features of apoptosis confirmed peroxynitrite-induced neuronal apoptosis. Total transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling-positive cells were counted in areas near and 0.2 mm away from the fiber track. The counts both peaked at 24 h with no significant difference between the two areas. However, at 6 and 12 h post-exposure the counts were significantly higher near than away from the fiber track (P=0.03 and P=0.007 respectively, paired t test). Immunohistochemical staining indicates caspase-3 was activated by peroxynitrite; this activation peaked at 6 h post-exposure, suggesting that activation of caspase-3 might be an early event in the apoptotic cell death cascade. We conclude that 1) peroxynitrite generated in the cord at the level produced by spinal cord injury induces neuronal apoptosis, indicating a role for peroxynitrite in secondary spinal cord injury; 2) caspase activation might be involved in peroxynitrite-induced neuronal apoptosis; 3) therefore removal of peroxynitrite should reduce secondary cell death after spinal cord injury.

We previously demonstrated that the concentration of peroxynitrite significantly increases following impact spinal cord injury (SCI). The aim of this study was to test whether the SCI-induced elevation of peroxynitrite induces neuronal death and consequent neurological deficits. Peroxynitrite was generated by administering 5 mM S-morpholinosydnonimine, a donor of peroxynitrite, through a microdialysis fiber into the gray matter of the rat spinal cord for 5 h. This mimics the concentration and duration of peroxynitrite elevation after SCI. Neuron death was assessed by counting the neurons along the fiber track in Cresyl Violet-stained sections removed at different times post-peroxynitrite exposure. Peroxynitrite induced significantly more neuron death than did the artificial cerebrospinal fluid (ACSF) control, with the percentage of neuronal loss being 17+/-2%, 28+/-2%, 39+/-3%, and 43+/-4% at 6, 12, 24 and 48 h post-peroxynitrite exposure (P=0.01

To determine whether reactive nitrogen species contribute to secondary damage in CNS injury, the time courses of nitric oxide, peroxynitrite, and nitrotyrosine production were measured following impact injury to the rat spinal cord. The concentration of nitric oxide measured by a nitric oxide-selective electrode dramatically increased immediately following injury and then quickly declined. Nitro-L-arginine reduced nitric oxide production. The extracellular concentration of peroxynitrite, measured by perfusing tyrosine through a microdialysis fiber into the cord and quantifying nitrotyrosine in the microdialysates, significantly increased after injury to 3.5 times the basal level, and superoxide dismutase and nitro-L-arginine completely blocked peroxynitrite production. Tyrosine nitration examined immunohistochemically significantly increased at 12 and 24 h postinjury, but not in sham-control sections. Mn(III) tetrakis(4-benzoic acid)-porphyrin (a novel cell-permeable superoxide dismutase mimetic) and nitro-L-arginine significantly reduced the numbers of nitrotyrosine-positive cells. Protein-bound nitrotyrosine was significantly higher in the injured tissue than in the sham-operated controls. These results demonstrate that traumatic injury increases nitric oxide and peroxynitrite production, thereby nitrating tyrosine, including protein-bound tyrosine. Together with our previous report that trauma increases superoxide, our results suggest that reactive nitrogen species cause secondary damage by nitrating protein through the pathway superoxide + nitric oxide peroxynitrite protein nitration.

We previously demonstrated that hydrogen peroxide concentration ([H2O2]) significantly increases after spinal cord injury (SCI). The present study explored (1) whether SCI-elevated [H2O2] is sufficient to induce oxidation and cell death, (2) if apoptosis is a pathway of H2O2-induced cell death, and (3) whether H2O2-induced oxidation and cell death could be reversed by treatment with the catalytic antioxidant Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP). H2O2 was perfused through a microcannula into the uninjured rat spinal cord to mimic the conditions induced by SCI. Protein and DNA oxidation, membrane phospholipids peroxidation (MLP), cell death and apoptosis were characterized by histochemical and immunohistochemical staining with antibodies against markers of oxidation and apoptosis. Stained cells were quantified in sections of H2O2-, or artificial cerebrospinal fluid (ACSF)-exposed with vehicle-, or MnTBAP-treated groups. Compared with ACSF-exposed animals, SCI-elevated [H2O2] significantly increased intracellular protein and DNA oxidation by threefold and MLP by eightfold in neurons, respectively. H2O2-elevated extracellular malondialdehyde was measured by microdialysis sampling. We demonstrated that SCI-elevated [H2O2] significantly increased extracellular malondialdehyde above pre-injury levels. H2O2 also significantly increased cell loss and the numbers of terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate-(dUTP)-biotin nick end labeling (TUNEL)-positive and active caspase-3-positive neurons by 2.3-, 2.8-, and 5.6-fold compared to ACSF controls, respectively. Our results directly and unequivocally demonstrate that SCI-elevated [H2O2] contributes to post-SCI MLP, protein, and DNA oxidation to induce cell death. Therefore, we conclude that (1) the role of H2O2 in secondary SCI is pro-oxidation and pro-cell death, (2) apoptosis is a pathway for SCI-elevated [H2O2] to induce cell death, (3) caspase activation is a mechanism of H2O2-induced apoptosis after SCI, and (4) MnTBAP treatment significantly decreased H2O2-induced oxidation, cell loss, and apoptosis to the levels of ACSF controls, further supporting MnTBAP's ability to scavenge H2O2 by in vivo evidence.

The present study explores in vivo whether and how prostaglandin F2α (PGF2α), a membrane phospholipid hydrolysis product, causes neuronal death. The concentration of PGF2α measured by microdialysis sampling increased threefold immediately following impact injury to the rat spinal cord. Administration of PGF2α into the cord through a dialysis fiber caused significant cell loss, increased extracellular levels of hydroxyl radicals and malondialdehyde – an end product of membrane lipid peroxidation – to 3.3 and 2.3 times basal levels, respectively. This suggests that PGF2α-induced cell death is partly due to hydroxyl radical-triggered peroxidation. Generating hydroxyl radical by administering Fenton's reagents into the cord through the fibers significantly increased malondialdehyde production – the first direct in vivo evidence that hydroxyl radical triggers membrane lipid peroxidation. Methylprednisolone significantly reduced the release of PGF2α upon spinal cord injury and blocked PGF2α-induced hydroxyl radical and malondialdehyde production, but did not significantly reduce Fenton's reagent-induced malondialdehyde production, despite the production of more malondialdehyde by PGF2α. This suggests that methylprednisolone may not directly scavenge hydroxyl radical, and that its ‘antioxidant’ effect is a consequence of blocking the pathways for producing toxic PGF2α and for PGF2α-induced hydroxyl radical formation, thereby reducing membrane lipid peroxidation.

To explore whether reactive oxygen species (ROS) play a role in the pathogenesis of amyotrophic lateral sclerosis (ALS), a unique microdialysis or microcannula sampling technique was used in mice transfected with a mutant Cu,Zn-superoxide dismutase (SOD1) gene from humans with familial ALS, mice transfected with the normal human SOD1 gene, and normal mice. We demonstrate for the first time that the levels of hydrogen peroxide (H(2)O(2)) and the hydroxyl radical ((.)OH) are significantly higher, and the level of the superoxide anion (O(2)(.-)) is significantly lower in ALS mutant mice than in controls, supporting by in vivo evidence the hypothesis that the mutant enzyme catalyzes (.)OH formation by the sequence: O(2)(.-) --H(2)O(2) --(.)OH. This removes doubts regarding the relevance of elevated ROS in FALS raised by in vitro experiments. The levels of oxidation products are also significantly higher in the mutant mice than in controls, consistent with some previous reports. Only the superoxide concentration differs between two controls among all the measurements. Our findings correlate in vivo a gene mutation to both elevated H(2)O(2) and (.)OH and increased oxidation of cellular constituents. The elevated H(2)O(2) in mutant mice indicates impairment of its detoxification pathways, perhaps by changed interactions between SOD1 and H(2)O(2) detoxification enzymes.-Liu, D., Wen, J., Liu, J., Li, L. The roles of free radicals in amyotrophic lateral sclerosis: reactive oxygen species and elevated oxidation of protein, DNA, and membrane phospholipids.

This work determines the ratio of DeltaH(vH) /DeltaH(cal) for staphylococcal nuclease (SN) denaturation in guanidine hydrochloride (GdnHCl) to test whether GdnHCl-induced denaturation is two-state. Heats of mixing of SN as a function of [GdnHCl] were determined at pH 7.0 and 25 degrees C. The resulting plot of DeltaH(mix) vs [GdnHCl] exhibits a sigmoid shaped curve with linear pre- and post-denaturational base lines. Extending the pre- and post-denaturational lines to zero [GdnHCl] gives a calorimetric DeltaH (DeltaH(cal)) of 24.1 +/- 1.0 kcal/mol, for SN denaturation in the limit of zero GdnHCl concentration. Guanidine hydrochloride-induced denaturation Gibbs energy changes in the limit of zero denaturant concentration (DeltaG degrees (N)(-)(D)) at pH 7. 0 were determined for SN from fluorescence measurements at fixed temperatures over the range from 15 to 35 degrees C. Analysis of the resulting temperature-dependent DeltaG degrees (N)(-)(D) data defines a van't Hoff denaturation enthalpy change (DeltaH(vH)) of 26. 4 +/- 2.8 kcal/mol. The model-dependent van't Hoff DeltaH(vH) divided by the model-independent DeltaH(cal) gives a ratio of 1.1 +/- 0.1 for DeltaH(vH)/DeltaH(cal), a result that rules out the presence of thermodynamically important intermediate states in the GdnHCl-induced denaturation of SN. The likelihood that GdnHCl-induced SN denaturation involves a special type of two-state denaturation, known as a variable two-state process, is discussed in terms of the thermodynamic implications of the process.

Damage caused by administering glutamate into the spinal cord was characterized histologically. Glutamate destroyed neurons for several hundred micrometers around the administering microdialysis fiber. At 24 h after treatment, significant (P=0.036) loss of neurons was observed (75%) relative to control (47%) near the fiber when glutamate was administered for 1 h at a concentration outside the fiber approximating the maximum glutamate released upon spinal cord injury. Significant loss of neurons (P=0.006, 0.022) was also caused by administering a combination of glutamate at about its average concentration released upon injury over the 1 h period of administration in combination with the maximum aspartate concentration released upon injury. This work provides a direct demonstration that the concentrations of excitatory amino acids released upon spinal cord injury are neurotoxic. The destruction of neurons by exposure to excitatory amino acids when there is also substantial loss of neurons simply from the presence of the microdialysis fiber may reflect sensitization of neurons to excitotoxicity by stress.

This paper reports a highly sensitive, specific, and reproducible method for the analysis of malondialdehyde (MDA) from microdialysates. The microdialysates were reacted with 2-thiobarbituric acid, and the TBA adducts were separated by HPLC and detected using a fluorescence detector. Butylated hydroxytoluene was used as an antioxidant to minimize formation of artifacts. The time course of MDA production following impact injury to the rat spinal cord was obtained using this improved method. MDA concentrations in the extracellular space gradually increased from a basal level of 20 ± 3.6 nM to 44 ± 18.1 nM during the first 2 hr, reached a maximum of 95 ± 19.8 nM at 5 hr, and then decreased to 36 ± 9.5 nM at 9 hr. The findings support the hypothesis that spinal cord injury leads to increased membrane lipid peroxidation.

To explore the time-dependent changes in the expressions of ferritin (Fn) and ferroportin (FPN) in macrophages infected with Mycobacterium (M.) tuberculosis of highly virulent strain (H37Rv strain) and Bacillus Calmette-Guerin (BCG) strain.RAW264.7 macrophages were respectively infected with M. tuberculosis H37Rv strain and BCG strain. At 1, 6, 12, 18 and 24 h after infection, the contents of Fn and FPN in the infected macrophage culture supernatants of the two groups were detected using ELISA, and meanwhile, the expression of Fn in the infected macrophages of each group was determined using Western blotting.After the macrophages were infected with different strains, the expression of Fn in macrophages increased as time went by; at 1 h after infection, the Fn expression in the control group was significantly higher than that in the H37Rv group and the BCG group; at 6 h after infection, the Fn expression in the two infected groups were higher than that in the control group; at 12 h, the Fn expression in the H37Rv group was higher than that in the control group, the difference was statistically significant (P0.05); at 18 and 24 h, the Fn expression was reduced in the order as H37Rv groupBCG groupcontrol group (P0.05). The expression of FPN decreased gradually as time went by. Compared with the normal control group, the expression in both H37Rv group and BCG group presented a lower level and H37Rv groupBCG groupcontrol group (P0.05).The infection of M.tuberculosis can lead to increased Fn expression and decreased FPN expression in macrophages, and the levels of Fn and FPN in the infected macrophages are both related to the strength of the virulence of M.tuberculosis.

Microdialysis was used to administer paraquat into the spinal cord of the anesthetized rat to determine the effects of the in vivo generation of the superoxide anion (O2.-) on neurons. Exposure to paraquat caused blockage of axonal conduction, destruction of the cell bodies of neurons, and a general release of amino acids. Thus, paraquat is quite harmful to neuronal tissue. Similarities between paraquat-induced damage and that previously observed from the hydroxyl radical are consistent with paraquat, causing damage by generation of reactive oxygen species and the widespread belief that generation of O2.- initiates the formation of destructive reactive oxygen species in a wide variety of traumas and other disorders.

Consultation research has focused especially on the process and implementation of consultation, the clinical outcomes of consultation, and the use of school psychology consultation by other school professionals. The present study used an empirically based scale designed to investigate the characteristics and behaviors of effective consultants as perceived by a large sample of classroom teachers (a) to determine their perceptions of the consultants' effectiveness, (b) to compare the factor structure of the scale with previous research, and (c) to investigate how the teachers' professional and demographic characteristics were related to their responses. The results indicated (a) that teachers found most of the consultant characteristics and behaviors that were rated to lie between important and extremely important on the scale's five-point rating scale; (b) that two factors (Consultant Knowledge, Process, and Application Skills; Consultant Interpersonal and Problem-Solving Skills) emerged from a factor analysis of the teachers' scale responses; (c) that these two factors compared favorably with previous research; and (d) that teachers' gender, age, highest degree earned, and years of teaching experience had significant impact on some of the scale responses. Limitations of the study and directions for future research also were addressed.

Free radicals and some free fatty acids, such as arachidonic acid metabolites, have been hypothesized to be contributors to secondary damage to the spinal cord upon injury. These two types of species may form a feedback loop in which generation of one type leads to formation of the other. In this study, to determine whether hydroxyl radical causes generation of arachidonic acid metabolites in vivo, we generated hydroxyl radical, a most reactive oxygen radical, in the rat spinal cord and measured resulting changes in levels of prostaglandin F 2α , an arachidonic acid metabolite that rises following traumatic injury. The hydroxyl radical was generated in the rat spinal cord by administering H 2 O 2 through one microdialysis fiber and FeC1 2 /EDTA through a parallel fiber. The prostaglandin F 2α in the collected microdialysates was measured by HPLC as its 3-bromomethyl-6,7-dimethoxy-l-methyl-2-(1H)-quinoxalinone derivative. Prostaglandin F 2α dramatically increased in response to hydroxyl radical generation, but declined substantially after 3 h of exposure. Prostaglandin F 2α was undetectable when either H 2 O 2 or FeC1 2 /EDTA was administered alone in control experiments, demonstrating that its formation was caused by generated hydroxyl radical.

We used an experimental model that we previously developed to characterize the damage caused by the agonists of glutamate receptors, N-methyl-D-aspartate (NMDA) and kainate, in the spinal cord in vivo, thereby testing further the utility of this model. Microdialysis was used to administer the toxins and to sample the release of other substances in response to these agents. The blockage of electrical conduction was monitored by recording the amplitudes of evoked potentials during administration of the damaging substances, and damage was assessed by postmortem histological examination. The released amino acids in microdialysates were measured by HPLC. Administration of 5 mM NMDA + 5 mM kainate into the gray matter blocked most postsynaptic responses and caused the release of amino acids. Administration of 10 mM NMDA and 10 mM kainate significantly destroyed cell bodies near the fiber. The advantage of this model is that histological, neurochemical, and electrophysiological parameters were obtained in the same experiment.

We developed a double microdialysis fiber technique to generate hydroxyl radicals (OH) in rat spinal cord. H 2 O 2 and FeCl 2 /EDTA were pumped through two parallel microdialysis fibers inserted into the spinal cord such that the reactants mix in the tissue to generate OH by the Fenton reaction. Generated OH was detected by administering phenylalanine through one fiber and measuring o -, m - and p -hydroxyphenylalanine in collected, dialysates by high pressure liquid chromatography and fluorescence detection. The hydroxyphenylalanines are produced by OH attacking the phenylalanine. OH generation was also accomplished in in vitro experiments and the results were consistent with in vivo experiments. This novel method to generate and measure OH radical in vivo overcomes difficulties in studying damage to tissue by short-lived OH. Although developed to study the role of OH in spinal cord injury, this method could be used to study other diseases involving OH damage.

A previously developed fluorescence method for determining leupeptin and the angiotensins is shown to be applicable to assaying other small arginine-containing peptides. In this procedure arginine residues are derivatized by reaction with benzoin. Femtomole sensivity is obtainable with this method. However, it cannot be used with larger arginine-containing peptides because those fragment upon derivatization.

Microdialysis sampling was used to characterize the release of norepinephrine and serotonin upon impact injury to the rat spinal cord. Increases in extracellular norepinephrine concentrations in response to injury were small and of short duration. In contrast, serotonin concentrations quickly rose 35-90 times following injury and took 30-45 min to return to control levels. Bleeding caused by injury was probably the major source of the increased serotonin levels. Our results allow a role for serotonin in secondary damage upon injury to the spinal cord but suggest that norepinephrine is not a very significant contributor to such damage.

Precursors to beta-endorphin (BE) and methionine enkephalin (ME), and proteolytic enzymes that cleave those BE and ME precursors to BE and ME, were determined in several milliliters of human cerebrospinal fluid. Endogenous peptides were purified by reversed-phase high-performance liquid chromatography (HPLC), and were detected with radioreceptor assay (RRA), radioimmunoassay, and mass spectrometry (MS). Total opioid receptor activity measurements and the profile of HPLC-receptor activity of human CSF samples were both used to monitor neuropeptide metabolism. MS data linked the molecular ion of ME to a unique fragment ion. A later-eluting fraction (84 min) in a 90-min HPLC gradient appeared in all HPLC-RRA profiles, contained opioid receptor activity that displaced [3H]etorphine, and the quantitative and qualitative patterns of opioid receptor activity in those profiles both changed within the few minutes that elapsed between acquiring the first and second cerebrospinal fluid samples. That 84-min fraction contained precursors to opioid peptides and was fractionated further with a more shallow 120-min HPLC gradient into three sections that displayed delta-opioid receptor-preferring activity, using [3H]ME as ligand. These three sections were hydrolyzed separately with human cerebrospinal fluid as the source for endogenous neuropeptides to yield products that correlated to immunoreactive BE in section I and immunoreactive ME in section III.

The metalloporphyrin Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) is a cell-permeable superoxide dismutase mimetic and a broad-spectrum scavenger of reactive species. Since MnTBAP may not cross the blood-brain barrier, this study evaluated the therapeutic potential of MnTBAP to treat spinal cord injury (SCI; 25 g x cm) by directly administering it into the intrathecal space of the rat spinal cord. The cells in spinal sections removed at 24 h post-SCI were immunohistochemically stained with anti-4-hydroxynonenal (HNE), a marker of membrane lipid peroxidation (MLP); anti-nitrotyrosine (Ntyr), a marker of protein nitration; and anti-neuron-specific enolase (NSE) antibodies. Immunostained neurons were counted for quantitative evaluation. Pre-treatment 30 min before SCI with 1 mg/kg MnTBAP or 4-h post-SCI treatment with 2.5 mg/kg MnTBAP administered into the intrathecal space significantly reduced MLP and protein nitration, and increased the number of surviving neurons compared to vehicle controls. However, post-SCI treatment with a standard regimen of methylprednisolone sodium succinate (MPSS; 30 mg/kg followed by 5.4 mg/kg for maintenance, iv administration), the only drug used for clinical treatment of SCI, not only did not reduce MLP and neuron loss, it increased protein nitration compared with vehicle controls (two-way analysis of variance [ANOVA] followed by the Tukey test). These results demonstrate that pre- and post-intrathecal treatments with the low doses of MnTBAP provide sustained neuroprotection by preventing oxidative stress and that post-treatment with MnTBAP is superior to post-treatment with MPSS in preventing oxidative stress and resulting neuron loss.

As part of our effort to study the role of reactive species in amyotrophic lateral sclerosis (ALS), the goal of this work is to explore the correlation between nitration and oxidation of proteins and mutation of Cu, Zn-superoxide dismutase (SOD1) in ALS. Transgenic mice overexpressing the mutant Cu, Zn-superoxide dismutase (mSOD1) gene from humans with familial ALS, wild-type mice overexpressing the normal human SOD1 gene and normal mice without gene overexpression were used. Brain sections from different regions of three groups of mice were double immunohistochemically stained with anti-neurofilament plus anti-nitrotyrosine or treated with 2,4-dinitrophenylhydrazine to label protein carbonyls, then double stained with anti-neurofilament plus anti-2,4-dinitrophenyl (anti-DNP). Neurons containing nitrated and oxidized proteins were visualized only in mSOD1 mice in the motor cortex, the cerebellar cortex and nucleus of hypoglossal nerves (regions related with movement). This correlates mutation of SOD1 to nitration and oxidation of neurons in the movement regions. By counting double-stained neurons, we demonstrated that the number of nitrotyrosine- and DNP-positive neurons was significantly higher in the brain sections of both motor and sensory cortex in mSOD1 mice than in the corresponding regions of control mice (P=0.005 to