Worldwide effects of non-native species on species-area relationships.

Non-native species have invaded most parts of the world, and the invasion process is expected to continue and accelerate. Because many invading non-native species are likely to become permanent inhabitants, future consideration of species-area relationships (SARs) should account for non-native species, either separately or jointly with native species. If non-native species occupy unused niches and space in invaded areas and extinction rate of native species remains low (especially for plants), the resultant SARs (with both native and non-native species) will likely be stronger. We used published and newly compiled data (35 data sets worldwide) to examine how species invasions affect SARs across selected taxonomic groups and diverse ecosystems around the world. We first examined the SARs for native, non-native, and all species. We then investigated with linear regression analyses and paired or unpaired t tests how degree of invasion (proportion of non-native species) affected postinvasion SARs. Postinvasion SARs for all species (native plus non-native) became significantly stronger as degree of invasion increased (r2 = 0.31, p = 0.0006), thus, reshaping SARs worldwide. Overall, native species still showed stronger and less variable SARs. Also, slopes for native species were steeper than for non-native species (0.298 vs. 0.153). There were some differences among non-native taxonomic groups in filling new niches (especially for birds) and between islands and mainland ecosystems. We also found evidence that invasions may increase equilibrial diversity. Study of such changing species-area curves may help determine the probability of future invasions and have practical implications for conservation.

Efectos Globales de las Especies No Nativas sobre las Relaciones Especie-Área Resumen Las especies no nativas han invadido la mayor parte del mundo y se espera que el proceso de invasión continúe y se acelere. Ya que muchas especies invasoras no nativas probablemente se conviertan en habitantes permanentes, la consideración a futuro de las relaciones especie-área (REA) debería considerar a las especies no nativas, ya sea por separado o en conjunto con las especies nativas. Si las especies no nativas ocupan nichos sin usar y el espacio en las áreas invadidas y la tasa de extinción de las especies nativas permanecen bajas (especialmente para las plantas), las REA resultantes (tanto con las especies nativas como las no nativas) probablemente sean más fuertes. Usamos datos publicados y recientemente compilados (35 conjuntos de datos mundiales) para examinar cómo las invasiones de especies afectan a las REA en grupos taxonómicos selectos y en diversos ecosistemas en todo el mundo. Primero examinamos las REA para todas las especies, así como para las nativas y las no nativas. Después investigamos con análisis de regresión lineal y pruebas t emparejadas o no emparejadas cómo afectó el grado de invasión (proporción de la especie no nativa) a las REA post-invasión. Las REA post-invasión para todas las especies (nativas más no nativas) se volvieron significativamente más fuertes conforme incrementó el grado de invasión (r2 = 0.31, p = 0.0006), remodelando así las REA en todo el mundo. En general, las especies nativas todavía mostraron REA más fuertes y menos variables. De igual manera, las pendientes de las especies nativas fueron más pronunciadas para las especies no nativas (0.298 vs. 0.153). Hubo algunas diferencias entre los grupos taxonómicos no nativos al llenar nichos nuevos (especialmente para las aves) y entre las islas y los ecosistemas de tierra firme. También encontramos evidencias de que las invasiones pueden incrementar la diversidad equilibrada. El estudio de dichas curvas cambiantes de relación especie-área podría ayudar a determinar la probabilidad de las futuras invasiones y tener implicaciones prácticas para la conservación.

Changes in taxonomic and phylogenetic diversity in the Anthropocene.

To better understand how ecosystems are changing, a multifaceted approach to measuring biodiversity that considers species richness (SR) and evolutionary history across spatial scales is needed. Here, we compiled 162 datasets for fish, bird and plant assemblages across the globe and measured how taxonomic and phylogenetic diversity changed at different spatial scales (within site α diversity and between sites spatial ß diversity). Biodiversity change is measured from these datasets in three ways: across land use gradients, from species lists, and through sampling of the same locations across two time periods. We found that local SR and phylogenetic α diversity (Faith's PD (phylogenetic diversity)) increased for all taxonomic groups. However, when measured with a metric that is independent of SR (phylogenetic species variation, PSV), phylogenetic α diversity declined for all taxonomic groups. Land use datasets showed declines in SR, Faith's PD and PSV. For all taxonomic groups and data types, spatial taxonomic and phylogenetic ß diversity decreased when measured with Sorensen dissimilarity and phylogenetic Sorensen dissimilarity, respectively, providing strong evidence of global biotic homogenization. The decoupling of α and ß diversity, as well as taxonomic and phylogenetic diversity, highlights the need for a broader perspective on contemporary biodiversity changes. Conservation and environmental policy decisions thus need to consider biodiversity beyond local SR to protect biodiversity and ecosystem services.

Determination of Postmortem Interval in Mice.

Despite the major use of mice in biomedical research, little information is available with regard to identifying their postmortem changes and using that information to determine the postmortem interval (PMI), defined as the time after death. Both PMI and environmental conditions influence decomposition (autolysis and putrefaction) and other postmortem changes. Severe decomposition compromises lesion interpretation and disease detection and wastes limited pathology resources. The goal of this study was to assess postmortem changes in mice in room temperature cage conditions and under refrigeration at 4 °C to develop gross criteria for the potential value of further gross and histologic evaluation. We used 108 experimentally naïve C57BL/6 mice that were humanely euthanized and then allocated them into 2 experimental groups for evaluation of postmortem change: room temperature (20 to 22 °C) or refrigeration (4 °C). PMI assessments, including gross changes and histologic scoring, were performed at hours 0, 4, 8, and 12 and on days 1 to 14. Factors such as temperature, humidity, ammonia in the cage, and weight change were also documented. Our data indicates that carcasses held at room temperature decomposed faster than refrigerated carcasses. For most tissues, decomposition was evident by 12 h at room temperature as compared with 5 d under refrigeration. At room temperature, gross changes were present by day 2 as compared with day 7 under refrigeration. Mice at room temperature lost 0.78% of their baseline body weight per day as compared with 0.06% for refrigerated mice (95% CI for difference 0.67% to 0.76%, P < 0.0005). This study supports the consideration of temperature and PMI as important factors affecting the suitability of postmortem tissues for gross and histologic evaluation and indicates that storage of carcasses under refrigeration will significantly slow autolysis.

Pattern and process of biotic homogenization in the New Pangaea.

Human activities have reorganized the earth's biota resulting in spatially disparate locales becoming more or less similar in species composition over time through the processes of biotic homogenization and biotic differentiation, respectively. Despite mounting evidence suggesting that this process may be widespread in both aquatic and terrestrial systems, past studies have predominantly focused on single taxonomic groups at a single spatial scale. Furthermore, change in pairwise similarity is itself dependent on two distinct processes, spatial turnover in species composition and changes in gradients of species richness. Most past research has failed to disentangle the effect of these two mechanisms on homogenization patterns. Here, we use recent statistical advances and collate a global database of homogenization studies (20 studies, 50 datasets) to provide the first global investigation of the homogenization process across major faunal and floral groups and elucidate the relative role of changes in species richness and turnover. We found evidence of homogenization (change in similarity ranging from -0.02 to 0.09) across nearly all taxonomic groups, spatial extent and grain sizes. Partitioning of change in pairwise similarity shows that overall change in community similarity is driven by changes in species richness. Our results show that biotic homogenization is truly a global phenomenon and put into question many of the ecological mechanisms invoked in previous studies to explain patterns of homogenization.

Physiologic and Behavioral Effects in Mice Anesthetized with Isoflurane in a Red-tinted or a Traditional Translucent Chamber.

Isoflurane has been characterized as a distressing agent for rodents, causing both physiologic and behavioral effects. Using a "darkened home cage" has been recommended during CO2 administration for rodent euthanasia; this is arguably a similar animal experience to anesthetic induction with isoflurane. Based on the premise that rodents perceive red light as darkness via the primary optic tract, we compared physiologic and behavioral markers of stress in 2 inbred strains of mice (C57BL/6J and BALB/cJ) anesthetized with isoflurane in either a red-tinted (dark) induction chamber or a traditional translucent induction chamber. Physiologic stress was assessed based on plasma levels of norepinephrine, epinephrine, and corticosterone. Stress-related behaviors (rearing, face wiping, and jumping) were recorded on video and scored from initiation of induction to loss of consciousness. No significant correlations were found between chamber type and physiologic stress hormones. As compared with the translucent chamber, stress-related behaviors were more frequent in the red-tinted chamber, including: 1) significantly higher rearing frequencies in BALB/cJ mice; 2) higher behavioral stress scores in BALB/cJ and male C57BL/6J mice; and 3) more face wiping behavior when considering all mice combined. These findings suggest that mice do not experience significant alleviation of physiologic indices of stress when anesthetized in a red-tinted induction chamber. Furthermore, isoflurane induction in the red-tinted chamber appeared to increase the expression of stress-related behaviors, particularly in BALB/cJ mice. Based on our findings and a growing body of literature on the unintended effects of red light, we do not recommend using red-tinted chambers for induction of anesthesia in mice.

Land snail dispersal, abundance and diversity on green roofs.

We present the first major systematic study of land snail diversity on green roofs. We surveyed 27 green roofs and the adjacent ground habitat in six major cities in the southeastern United States. We found a total of 18 species of land snails, with three considered to be non-native or invasive species. The majority of land snails encountered in surveys are widespread, generalist species, typically adapted to open habitats. Twelve of the land snails encountered are "greenhouse" species that are very commonly transported via the horticultural trade. Therefore, we infer that at least some land snail species are introduced to green roofs via initial green roof installation and associated landscaping. The major determinants of snail species richness and abundance are the size of each roof and the quality of green roof maintenance regime.

Assessing the value of ureteral stent placement in pediatric kidney transplant recipients.

BACKGROUND: Ureteral stent placement at kidney transplantation may reduce stenosis or leakage (S/L) complication rates. However, stent placement may also increase risk for early urinary tract infection (early UTI; <3 months after transplant) and BK virus allograft nephropathy (BKVAN). In children, the usefulness of stent placement is not well defined. METHODS: We analyzed retrospective data from children transplanted at our center for the three above outcomes in relation to stents. At our center, stent placement decision is driven by surgeon preference. RESULTS: Among 129 transplants from 1996 to 2006, early UTI was seen in 9.3% and S/L in 4.6%. By univariate analyses, stent placement was a significant risk factor for early UTI (P=0.0399) but not protective for S/L (P=0.23). In multivariate analyses, stent placement, human leukocyte antigen match, and bladder augmentation increased the odds ratio for early UTI. Only deceased donor source increased the odds ratio for S/L. In a truncated data set from 1999 to 2006, BKVAN occurred in 9 of 93 (9.6%). Per minute increase in warm ischemia time was the only significant risk factor for BKVAN by both univariate and Cox regression analyses. Stent placement did not improve graft survival (P=0.5726) but required general anesthesia for removal in the operating room, leading to additional cost and potential risk. CONCLUSION: Routine stent placement in children in this era of low urological complication rates and BKVAN needs reevaluation.

Altered expression of zonula occludens-2 precedes increased blood-brain barrier permeability in a murine model of fulminant hepatic failure.

Brain edema secondary to increased blood-brain barrier (BBB) permeability is a lethal complication in fulminant hepatic failure (FHF). Intact tight junctions (TJ) between brain capillary endothelial cells are critical for normal BBB function. However, the role of TJ in FHF has not been explored. We hypothesized that alterations in the composition of TJ proteins would result in increased BBB permeability in FHF. In this study, FHF was induced in C57BL/6J mice by using azoxymethane. BBB permeability was assessed with sodium fluorescein. Expression of TJ proteins was determined by Western blot, and their cellular distribution was examined using immunofluorescent microscopy. Comatose FHF mice had significant cerebral sodium fluorescein extravasation compared with control and precoma FHF mice, indicating increased BBB permeability. Western blot analysis showed a significant decrease in zonula occludens (ZO)-2 expression starting in the precoma stage. Immunofluorescent microscopy showed a significantly altered distribution pattern of ZO-2 in isolated microvessels from precoma FHF mice. These changes were more prominent in comatose FHF animals. Significant alterations in ZO-2 expression and distribution in the tight junctions preceded the increased BBB permeability in FHF mice. These results suggest that ZO-2 may play an important role in the pathogenesis of brain edema in FHF.

Establishment of cholinergic neuron-like cell lines with differential vulnerability to nitrosative stress.

Cholinergic cell lines were established by fusion of embryonic day 17 wild-type neurons from rat basal forebrain (BF) and upper brainstem (BS) with N18tg neuroblastoma cells. Isolated clones expressed choline acetyltransferase (ChAT) and neuronal nitric oxide synthase (nNOS) activities that were increased upon differentiation with retinoic acid. Clones from the BF expressed high levels of the tyrosine kinase type A (TrkA) receptor expression and activation of the mitogen-activated kinase ERK2 upon treatment with nerve growth factor. Like wild-type cholinergic populations, the six clones studied were variably resistant to nitric oxide (NO) excess from addition of S-nitroso-N-acetyl-D, L-penicillamine (SNAP). Of these, the BS2 clone exhibited resistance like in vivo BS cholinergic neurons, while the MS10 clone mimicked in vivo BF vulnerability. Apoptosis in response to NO excess was preceded by increases in mitochondrial responses bax/bcl-2 ratios, but cytochrome C was not released. Mitochondrial levels of apoptosis initiating factor (AIF) were either unchanged or increased, and only in MS clones was endonuclease G (EndoG) released. Microarray data indicated the existence of endoplasmic reticular (ER) stress and caspase-4 and caspase-12 were involved in the pathway to DNA fragmentation. The array data also indicated a survival role for mdm2, and its blockade rendered vulnerable the brainstem survivor clone BS2. Akt and ERK1/2 pathways were activated in response to NO and their blockade increased DNA fragmentation. Blockade of GSK-3 alpha/beta, a downstream target of Akt, reduced SNAP toxicity and this was more prominent in basal forebrain clones. We have identified two cholinergic cell lines useful for molecular studies of cholinergic vulnerability. We hypothesize that, in cholinergic neurons, control of ER stress signaling may be a major factor in differential vulnerability.

The botanist effect revisited: plant species richness, county area, and human population size in the United States.

The "botanist effect" is thought to be the reason for higher plant species richness in areas where botanists are disproportionately present as an artefactual consequence of a more thorough sampling. We examined whether this was the case for U.S. counties. We collated the number of species of vascular plants, human population size, and the area of U.S. counties. Controlling for spatial autocorrelation and county area, plant species richness increased with human population size and density in counties with and without universities and/or botanical gardens, with no significant differences in the relation between the two subsets. This is consistent with previous findings and further evidence of a broad-scale positive correlation between species richness and human population presence, which has important consequences for the experience of nature by inhabitants of densely populated regions. Combined with the many reports of a negative correlation between the two variables at a local scale, the positive relation between plant species richness in U.S. counties and human population presence stresses the need for the conservation of seminatural areas in urbanized ecosystems and for the containment of urban and suburban sprawl.

Aging causes partial loss of basal forebrain but no loss of pontine reticular cholinergic neurons.

Cholinergic degeneration occurs in several neurodegenerative diseases. To investigate whether normal aging causes selective neurodegeneration, we compared counts of cholinergic neurons in the medial septum/vertical limb of the diagonal band and pedunculopontine and laterodorsal tegmental nuclei of the brainstem in young and aged Long-Evans rats characterized for their spatial learning ability in the Morris water maze. A subset of aged rats (aged-unimpaired) learned the spatial learning task as young rats, whereas another group (age-impaired) showed poorer learning than young animals. In the medial septum/diagonal band, there was a significant loss (-23%, P < 0.02) of cholinergic neurons in aged-impaired animals compared with young subjects. In the brainstem, there were no significant differences in cholinergic cell number in any group. This selective loss of cholinergic neurons may, in part, account for the cognitive deficits observed in aging and, considering previous findings in this model, may be related to oxidative stress.

Aging is neuroprotective during global ischemia but leads to increased caspase-3 and apoptotic activity in hippocampal neurons.

Previously, we found a significantly greater number of surviving CA1 neurons to global ischemia in the aged (24-month-old) F344 rats than in young (4-month-old) rats. The present study tests the hypothesis that aging retards neuronal death in the hippocampal CA1 region following cerebral ischemia. The CA1 "living cell ratio" was significantly greater in aged than in young rats at three days (62+/-8% vs. 30+/-8%) and at eight days (36+/-6% vs. 17+/-5%), but not at 14 days (15+/-12% vs. 18+/-12%) following ischemia. The number of the CA1 cells exhibiting co-localized TdT-mediated X-dUTP nick end labeling reaction and caspase-3 active peptide (C3AP) immunoreactivity was greater in aged than young animals at three and eight days following ischemia (36+/-8/mm vs. 3+/-1/mm and 36+/-14 vs. 0+/-0, p<0.05 respectively). Also, the total number of C3AP-positive cells in the CA1 region in the aged group was significantly greater than in the young group at three and eight days post-ischemia (p<0.05). Aging appears to delay caspase-3-dependent apoptotic cell death induced by global ischemia in the CA1 region of the hippocampus, consistent with an age-induced neuroprotective process.

Brain cholinergic vulnerability: relevance to behavior and disease.

The major populations of cholinergic neurons in the brain include two "projection" systems, located in the pontine reticular formation and in the basal forebrain. These two complexes comprise, in part, the anatomical substrates for the "ascending reticular activating system" (ARAS). The pontine cholinergic system relays its rostral influences mainly through thalamic intralaminar nuclei, but it also connects to the basal forebrain and provides a minor innervation of cortex. The basal forebrain cholinergic complex (BFCC) projects directly to cortex and hippocampus, and has a minor connection with the thalamus. Recent data reveal that a parallel system of basal forebrain GABAergic projection neurons innervates cortex/hippocampus in a way that seems to complement the BFCC. Generally, the picture developed from more than 50 years of research is consistent with a "global" influence of these two ascending cholinergic projections on cortical and hippocampal regions. Seemingly, the BFCC acts in tandem or in parallel with the pontine cholinergic projection to activate the electro-encephalogram, increase cerebral blood flow, regulate sleep-wake cycling, and modulate cognitive function. There are quite a number and variety of human brain conditions, notably including Alzheimer's disease, in which degeneration of basal forebrain cholinergic neurons has been documented. Whether the corticopetal GABA system is affected by disease has not been established. Studies of degeneration of the pontine projection are limited, but the available data suggest that it is relatively preserved in Alzheimer's disease. Hypotheses of BFCC degeneration include growth factor deprivation, intracellular calcium dysfunction, amyloid excess, inflammation, and mitochondrial abnormalities/oxidative stress. But, despite considerable research conducted over several decades, the exact mechanisms underlying brain cholinergic vulnerability in human disease remain unclear.

Hippocampal progenitor cells express nestin following cerebral ischemia in rats.

This study addresses whether hippocampal progenitor cells express nestin following cerebral ischemia in rats. Cell counts within the hippocampal hilus were significantly greater following severe (eight-vessel occlusion) ischemia than following intermediate (four-vessel occlusion) ischemia (1527+/-87/mm2 vs. 918+/-71/mm2). Bromedeoxyuridine-positive cell counts were significantly higher with severe ischemia than with intermediate ischemia or in sham-operated animals, respectively (368+/-45, 43+/-14 and 7+/-1/mm2). In the eight-vessel occlusion group, 47+/-8/mm2 bromedeoxyuridine-labeled cells expressed nestin, significantly higher than in the four-vessel occlusion group and sham-operated animals (1+/-1 and 1+/-0/mm2, P<0.01 vs. eight-vessel occlusion, respectively). Confocal microscopy verified that a subset of the bromedeoxyuridine-positive cells expressed nestin. In conclusion, severe ischemia elicits nestin expression in hippocampal progenitor cells in rats.

Aging blunts ischemic-preconditioning-induced neuroprotection following transient global ischemia in rats.

The present study examines the hypothesis that aging defined by the 50% survival age compromises neuroprotection afforded by ischemic preconditioning (IPC). Sixty-four male F344 rats aged 4- and 24-months, respectively, were subjected to IPC, (3-min ischemia) or sham-surgery followed by 10-min (full) ischemia or sham-surgery 2 days later. There were 4 groups at each age: sham-surgery-sham-surgery (SS), preconditioning-sham-surgery (PS), preconditioning-ischemia (PI) and sham-surgery-ischemia (SI) groups. Assessments of histology and immunoreactivities of N-methyl-D-aspartic acid receptor 1 (NMDAr1) and caspase-3 active peptide (C3AP) in the hippocampal CA1 region were performed 8 days after full ischemia. The CA1 "living cell ratio" was greater in the aged SI group than in the young SI group (32+/-6% vs. 17+/-5%, p<0.05), whereas the degree of protection against full ischemia afforded by IPC was reduced in the aged compared with the young (53+/-17% vs. 241+/-25%, P<0.0001). The basal level of NMDAr1 immunofluorescence was significantly higher in young animals, while the numbers of C3AP-positive cells were greater in all three aged ischemic groups as compared to respective young groups (p<0.01, p=0.055 and p<0.05). A fourth method of assessing cell damage using Fluoro Jade C labeled degenerating neurons that were also intensively eosinophilic. Counts of Fluoro Jade C-positive cells were higher in the young SI group than in the aged SI group (P<0.05), suggesting that mechanisms of ischemic cell death may change with aging. In conclusion, aging alters mechanisms of ischemic cell death in CA1 neurons and ischemic tolerance mechanisms are blunted by aging.

Pontine cholinergic neurons depend on three neuroprotection systems to resist nitrosative stress.

Brainstem cholinergic populations survive in neurodegenerative disease, while basal forebrain cholinergic neurons degenerate. We have postulated that variable resistance to oxidative stress may in part explain this. Rat primary cultures were used to study the effects of several nitrosative/oxidative stressors on brainstem (upper pons, containing pedunculopontine and lateraldorsal tegmental nuclei; BS) cholinergic neurons, comparing them with medial septal (MS), and striatal cholinergic neurons. BS cholinergic neurons were significantly more resistant to S-nitro-N-acetyl-d,l-penicillamine (SNAP), sodium nitroprusside (SNP), and hydrogen peroxide than were MS cholinergic neurons, which in turn were more resistant than striatal cholinergic neurons. Pharmacological analyses using specific inhibitors of neuroprotective systems also revealed differences between these three cholinergic populations with respect to their vulnerability to SNAP. Toxicity of SNAP to BS neurons was exacerbated by blocking NF-kappaB activation with SN50 or ERK1/2 activation by PD98059, or by inhibition of phosphoinositide-3 kinase (PI3K) activity by LY294002. In contrast, SNAP toxicity to MS neurons was augmented only by SN50, and SNAP toxicity to striatal cholinergic neurons was not increased by any of these three pharmacological agents. In neuron-enriched primary cultures, BS cholinergic neurons remained resistant to SNAP while MS cholinergic neurons remained vulnerable to this agent. Immunohistochemical experiments demonstrated nitric oxide (NO)-induced increases in nuclear levels of phospho-epitopes for ERK1/2 and Akt, and of the p65 subunit of NF-kappaB, within BS cholinergic neurons. These data indicate that the relative resistance of BS cholinergic neurons to toxic levels of nitric oxide involves three intrinsic neuroprotective pathways that control transcriptional and anti-apoptotic cellular functions.

The role of glutathione in nitric oxide donor toxicity to SN56 cholinergic neuron-like cells.

Our study was designed to determine if compounds used experimentally to generate nitric oxide excess differ in ability to elicit degenerative stress to cholinergic neurons and, if so, what mechanisms account for their differences. Nitric oxide donors are often used experimentally in attempts to emulate the bioactivities of endogenous NO, but the pharmacological actions of NO donors can vary dramatically according to the species of NO (NOx) and other agents (e.g., iron cations, cyanide anion, superoxide anion) released, and as affected by the state of the cellular redox environment. To determine whether different types of NO donors exert differential toxicity in a cholinergic neuronal model, we measured cell viability markers, indicators of NOx formation, levels of intracellular-reduced glutathione (GSH), protein nitrosothiols, and the activation of the transcription factor NF-kappaB in a mouse medial septal cholinergic cell line (clone SN56) following exposure to the NO donors S-nitroso-N-acetyl-dl-penicillamine (SNAP), 3-morpholinosydnonimine (SIN-1), or sodium nitroprusside (SNP). SNAP and SIN-1, but not SNP, elicited dramatic increases in media nitrite and intracellular NOx-related fluorescence from cells preloaded with a NOx indicator. Nevertheless, SN56 cells were readily killed by SNP (IC(50) approximately 0.5 mM), while even higher levels (up to 2 mM) of SNAP or SIN-1 were essentially ineffective. SNAP (an NO(+) generator) and SIN-1 (a peroxynitrite generator) both caused increases in SN56 GSH levels; in contrast, SNP caused an immediate and rapid decline in GSH. The increase in GSH in response to SNAP and SIN-1 probably indicates augmentation of intracellular defense mechanisms, because prior depletion of GSH rendered the cells vulnerable to these two donors. GSH depletion did not change the potency of SNP, but GSH depletion made SNAP about twice as potent as SNP. SNAP and SNP, but not SIN-1, activated the transcription factor NF-kappaB, as indicated by increases in p65 nuclear immunoreactivity. Treatment with SNAP, but not SNP or SIN-1, increased levels of S-nitrosothiols in SN56 proteins, consistent with the transfer of an NO(+) equivalent to intracellular thiols. Our experiments show that these three NO donors differ dramatically in their ability to intoxicate SN56 cells, probably because of the different species of NOx and other agents they release, and as reflected in their differing modes of interaction with cellular antioxidant and survival systems.

Increased severity of acute cerebral ischemic injury correlates with enhanced stem cell induction as well as with predictive behavioral profiling.

An 8-vessel-occlusion (8VO) method was developed to compare with the conventional 4-vessel-occlusion (4VO) in hippocampal ischemic damage and progenitor cell induction 10 days following ischemia in female rats. Eight posture-relevant tests were performed following ischemia to correlate grades of postural abnormality with the histological outcome. The total hippocampal living cell ratio including 7 hippocampal subregions in 8VO group (n=11) was much lower than that in 4VO group (n=10, 51+/-5% vs. 78+/-4, p<0.01). In 4VO group, BrdU positive cells were mainly located in the subgranular zone (SGZ) with a count of 54+/-20/mm2 (7micro-thick slice), comparable to the maximal level following global ischemia in male gerbils and rats reported so far referring to slice-thickness differences (50-60 micro-thick slices). Similarly, nestin-bearing cells were 29+/-11/ mm2. In 8VO group, BrdU and nestin positive cells increased by 10 times. Triple staining of BrdU, nestin and DAPI demonstrated that BrdU-immunoreactivity was extensively distributed in the hippocampal hilus while the nestin was mainly located along the SGZ. Most of nestin labeling was not co-localized with the BrdU, indicating that establishment of these cells might precede BrdU injections (8 and 9d post ischemia). Behavioral scores were much greater for 8VO group than for 4VO group and composite postural scores well correlated with the hippocampal cell loss. In conclusion, severe ischemia correlates with vigorous induction of the hippocampal progenitor cells in rats while behavioral profiling of posture changes permits prediction of severity of damage.

Atmospheric pressure ionization LC-MS-MS determination of urushiol congeners.

This paper describes atmospheric pressure ionization (API) LC-MS-MS determination of urushiols, 3-n-alkenyl- and -alkyl-substituted catechols responsible for poison oak dermatitis. Urushiol was isolated from Western poison oak according to the method of Elsohly et al. (1) (J. Nat. Prod. 1982, 45, 532-538)-the purified preparation contained C(17)- and C(15)-substituted urushiols with zero, one, two, and three double bonds as determined from GC-MS analysis of trimethylsilyl derivatives. Urushiol mixtures were separated on a C(18) reversed phase HPLC column with a methanol-water gradient with urushiols eluting in 100% methanol. Atmospheric pressure chemical ionization (APCI) produced primarily [M - H](-) and MH(+) molecule ions. Electrospray ionization (ESI) yielded [M - H](-) and adduct ions including [M + Cl](-). Daughter ions of [M - H](-) included quinoid radical anions ([M - H - H(2)](-) and m/z 122(-)) and a benzofuran phenate (m/z 135(-)). A suite of hydrocarbon fragments were produced by collision-induced dissociation of MH(+) directly or via an intermediate [MH - H(2)O](+) daughter ion. Six urushiol congeners, one not previously reported in poison oak, were determined by negative ion API-LC-MS-MS with detection limits of approximately 8 pg/microL (ESI) and approximately 800 pg/microL (APCI). API-LC-MS-MS was used to determine urushiol in surface wipes, air samples, and plant materials.

PBDEs in the San Francisco Bay Area: measurements in harbor seal blubber and human breast adipose tissue.

To explore the levels of polybrominated diphenyl ethers (PBDEs) in California, samples from 11 archived harbor seals (Phoca vitulina Richardsi) from the San Francisco Bay and breast adipose tissue samples from 23 women were analyzed. The levels of PBDEs in human tissue samples were in the low ng/g fat range, with PBDEs 47, 153, 154, 99, and 100 as the major congeners. Average sigma PBDEs (86 ng/g fat) in these California women are the highest human levels reported to date. An inverse relationship between concentration of PBDEs and age of these women was apparent. The levels of PBDEs measured in harbor seal blubber were in the low ng/g to low microg/g fat range, with the same major congeners as those measured in the human tissues. PBDE 47 was the highest among all congeners measured in both human tissue and seal blubber samples. The concentrations of PBDEs in harbor seals in the San Francisco Bay have increased dramatically over the past decade, with current levels among the highest reported for this species.