The Abundance of C2H4 in the Circumstellar Envelope of IRC+10216.

High spectral resolution mid-IR observations of ethylene (C2H4) towards the AGB star IRC+10216 were obtained using the Texas Echelon Cross Echelle Spectrograph (TEXES) at the NASA Infrared Telescope Facility (IRTF). Eighty ro-vibrational lines from the 10.5 µm vibrational mode ν7 with J ≲ 30 were detected in absorption. The observed lines are divided into two groups with rotational temperatures of 105 and 400 K (warm and hot lines). The warm lines peak at ≃ -14 km s-1 with respect to the systemic velocity, suggesting that they are mostly formed outwards from ≃ 20R⋆. The hot lines are centered at -10 km s-1 indicating that they come from a shell between 10 and 20R⋆. 35% of the observed lines are unblended and can be fitted with a code developed to model the emission of a spherically symmetric circumstellar envelope. The analysis of several scenarios reveal that the C2H4 abundance relative to H2 in the range 5 - 20R⋆ is 6.9 × 10-8 in average and it could be as high as 1.1 × 10-7. Beyond 20R⋆, it is 8.2 × 10-8. The total column density is (6.5 ± 3.0) × 1015 cm-2. C2H4 is found to be rotationally under local thermodynamical equilibrium (LTE) and vibrationally out of LTE. One of the scenarios that best reproduce the observations suggests that up to 25% of the C2H4 molecules at 20R⋆ could condense onto dust grains. This possible depletion would not influence significantly the gas acceleration although it could play a role in the surface chemistry on the dust grains.

Progressive dopaminergic alterations and mitochondrial abnormalities in LRRK2 G2019S knock-in mice.

Mutations in the LRRK2 gene represent the most common genetic cause of late onset Parkinson's disease. The physiological and pathological roles of LRRK2 are yet to be fully determined but evidence points towards LRRK2 mutations causing a gain in kinase function, impacting on neuronal maintenance, vesicular dynamics and neurotransmitter release. To explore the role of physiological levels of mutant LRRK2, we created knock-in (KI) mice harboring the most common LRRK2 mutation G2019S in their own genome. We have performed comprehensive dopaminergic, behavioral and neuropathological analyses in this model up to 24months of age. We find elevated kinase activity in the brain of both heterozygous and homozygous mice. Although normal at 6months, by 12months of age, basal and pharmacologically induced extracellular release of dopamine is impaired in both heterozygous and homozygous mice, corroborating previous findings in transgenic models over-expressing mutant LRRK2. Via in vivo microdialysis measurement of basal and drug-evoked extracellular release of dopamine and its metabolites, our findings indicate that exocytotic release from the vesicular pool is impaired. Furthermore, profound mitochondrial abnormalities are evident in the striatum of older homozygous G2019S KI mice, which are consistent with mitochondrial fission arrest. We anticipate that this G2019S mouse line will be a useful pre-clinical model for further evaluation of early mechanistic events in LRRK2 pathogenesis and for second-hit approaches to model disease progression.

Adult neurogenesis and neurite outgrowth are impaired in LRRK2 G2019S mice.

The generation and maturation of adult neural stem/progenitor cells are impaired in many neurodegenerative diseases, among them is Parkinson's disease (PD). In mammals, including humans, adult neurogenesis is a lifelong feature of cellular brain plasticity in the hippocampal dentate gyrus (DG) and in the subventricular zone (SVZ)/olfactory bulb system. Hyposmia, depression, and anxiety are early non-motor symptoms in PD. There are parallels between brain regions associated with non-motor symptoms in PD and neurogenic regions. In autosomal dominant PD, mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are frequent. LRRK2 homologs in non-vertebrate systems play an important role in chemotaxis, cell polarity, and neurite arborization. We investigated adult neurogenesis and the neurite development of new neurons in the DG and SVZ/olfactory bulb system in bacterial artificial chromosome (BAC) human Lrrk2 G2019S transgenic mice. We report that mutant human Lrrk2 is highly expressed in the hippocampus in the DG and the SVZ of adult Lrrk2 G2019S mice. Proliferation of newly generated cells is significantly decreased and survival of newly generated neurons in the DG and olfactory bulb is also severely impaired. In addition, after stereotactic injection of a GFP retrovirus, newly generated neurons in the DG of Lrrk2 G2019S mice exhibited reduced dendritic arborization and fewer spines. This loss in mature, developed spines might point towards a decrease in synaptic connectivity. Interestingly, physical activity partially reverses the decrease in neuroblasts observed in Lrrk2 G2010S mice. These data further support a role for Lrrk2 in neuronal morphogenesis and provide new insights into the role of Lrrk2 in adult neurogenesis.

Impaired dopaminergic neurotransmission and microtubule-associated protein tau alterations in human LRRK2 transgenic mice.

Mutations in the Leucine Rich Repeat Kinase 2 (LRRK2) gene, first described in 2004 have now emerged as the most important genetic finding in both autosomal dominant and sporadic Parkinson's disease (PD). While a formidable research effort has ensued since the initial gene discovery, little is known of either the normal or the pathological role of LRRK2. We have created lines of mice that express human wild-type (hWT) or G2019S Lrrk2 via bacterial artificial chromosome (BAC) transgenesis. In vivo analysis of the dopaminergic system revealed abnormal dopamine neurotransmission in both hWT and G2019S transgenic mice evidenced by a decrease in extra-cellular dopamine levels, which was detected without pharmacological manipulation. Immunopathological analysis revealed changes in localization and increased phosphorylation of microtubule binding protein tau in G2019S mice. Quantitative biochemical analysis confirmed the presence of differential phospho-tau species in G2019S mice but surprisingly, upon dephosphorylation the tau isoform banding pattern in G2019S mice remained altered. This suggests that other post-translational modifications of tau occur in G2019S mice. We hypothesize that Lrrk2 may impact on tau processing which subsequently leads to increased phosphorylation. Our models will be useful for further understanding of the mechanistic actions of LRRK2 and future therapeutic screening.

A comparative analysis of leucine-rich repeat kinase 2 (Lrrk2) expression in mouse brain and Lewy body disease.

Pathogenic substitutions in leucine-rich repeat kinase 2 (LRRK2, Lrrk2) have been genetically linked to familial, late-onset Parkinsonism. End-stage disease is predominantly associated with nigral neuronal loss and Lewy body pathology, but patients may have gliosis, tau or ubiquitin inclusions (pleomorphic pathology). The anatomical distribution of Lrrk2 protein may provide insight into its function in health and neurodegeneration, thus we performed a comparative study with 'in-house' and commercially available Lrrk2 antibodies using brain tissue from wild type and human Lrrk2 transgenic bacterial artificial chromosome (BAC) mice, and from diffuse Lewy body disease (DLBD) patients. Lrrk2 protein was ubiquitously expressed and relatively abundant in most brain regions, including the substantia nigra, thalamus and striatum. Lrrk2 was not a major component of Lewy body or neuritic pathology associated with Parkinson's disease. However, selective loss of dopaminergic neurons in Lrrk2-associated Parkinsonism argues the protein may have regional-specific interactions. Lrrk2 immunohistochemical staining was present in the subventricular zone, a region containing stem cells that give rise to both neurons and glia. A role for Lrrk2 in neurogenesis might provide further insight into the aberrant role of mutant protein in age-associated neurodegeneration with pleomorphic pathology.

Antitumour efficacy of VEGFR2 tyrosine kinase inhibitor correlates with expression of VEGF and its receptor VEGFR2 in tumour models.

During the development of indazolylpyrimidines as novel and potent inhibitors of vascular endothelial growth factor (VEGF) receptor-2 (VEGFR2) tyrosine kinase, we observed that some human tumour xenografts are more sensitive to VEGFR2 kinase inhibitors than others. A better understanding of the basis for this differential response may help to identify a predictive marker that would greatly aid in the identification of a suitable patient population for treatment. One representative compound from the indazolylpyrimidine series is GW654652 that inhibited all three VEGFRs with similar potency. The inhibition of VEGFR2 kinase by GW654652 was about 150 to >8800 more potent than the inhibition of eight other kinases tested. GW654652 inhibited VEGF- and bFGF-induced proliferation in endothelial cells with an IC(50) of 110 and 1980 nM, respectively, and has good pharmacokinetic profile in mouse and dog. We investigated the association between VEGF and VEGFR2 expression and the antitumour efficacy of GW654652, in various xenograft models. Statistically significant associations were observed between the antitumour efficacy of GW654652 in xenografts and VEGF protein (P=0.005) and VEGFR2 expression (P=0.041). The oral dose of GW654652 producing 50% inhibition of tumour growth (ED(50)) decreased with increasing levels of VEGF (r=-0.94); and, in contrast, the ED(50) increased with the increased expression of VEGFR2 (r=0.82). These results are consistent with the observed inverse correlation between VEGF and VEGFR2 expression in tumours. These findings support the hypothesis that VEGF and VEGFR2 expression by tumours may predict the therapeutic outcome of VEGFR kinase inhibitors.

The rational design of potential chemotherapeutic agents: synthesis of bryostatin analogues.

The bryostatins are a unique family of cancer chemotherapeutic candidates isolated from marine bryozoa. While their molecular mode of action is not known, these macrolactones exhibit high affinities for protein kinase C (PKC) isozymes, compete for the phorbol ester binding site on PKC, and stimulate kinase activity in vitro and in vivo. Unlike the phorbol esters, they do not act as tumor promoters. Despite promising biological properties, the supply of these compounds is limited by the difficulty of their isolation from natural sources and their synthetic complexity. A new class of bryostatin analogues which retain the putative recognition domain of the bryostatins but are simplified through deletions and modifications in the C1-C14 spacer domain have been designed using computer models. A convergent synthesis has been realized for the production, in gram quantities, of these recognition and spacer domains whose coupling allows for the generation of a range of analogues. The final closure process involves a novel macrotransacetalization reaction which proceeds with complete stereoselectivity. The solution structures of two synthetic analogues were determined by NMR spectroscopy and found to be very similar to the previously reported structures of bryostatins 1 and 10. In addition, these structures appear to indicate that the stereochemistry of the C3 hydroxyl group plays a significant role in the conformation of the macrolactone. All analogues bound strongly to a mixture of PKC isozymes, and several exhibited significant levels of in vitro growth inhibitory activity against human cancer cell lines. Taken together, this work provides important steps toward the development and understanding of simplified, synthetically accessible analogues of the bryostatins as potential chemotherapeutic agents.

Lessons from genetically engineered animal models. III. Lessons learned from gastrin gene deletion in mice.

Gastrin is the principal hormonal inducer of gastric acid secretion. Chronic hypergastrinemia, leading to hypersecretion of gastric acid and increased proliferation of parietal and enterochromaffin-like (ECL) cells, has been well described. In contrast, the physiological consequences of chronic gastrin deficiency had been poorly understood until the recent genetic engineering of mouse mutants containing a gastrin gene deletion by homologous recombination in embryonic stem cells. This themes article describes the consequences of constitutive gastrin deficiency on the development and physiology of the stomach. A lack of gastrin disrupts basal gastric acid secretion and renders the acid secretory system unresponsive to acute histaminergic, cholinergic, and gastrinergic stimulation. The defect in acid secretion is greater than would have been predicted from previous studies in which gastrin action was acutely blocked. Cellular changes include thinning of the gastric mucosa in the gastrin-deficient mice, with a reduction in parietal cells and reduced expression of markers of parietal and ECL cell-differentiated functions. The results suggest that gastrin is required for the functional maturation of the acid-secretory system.

Evaluation of activity of selected ophthalmic antimicrobial agents in combination against common ocular microorganisms.

OBJECTIVE: To determine in vitro efficacy of gentamicin, tobramycin, and miconazole when used in combination, with or without atropine, against Pseudomonas or Aspergillus sp. PROCEDURE: Selected ophthalmic agents were combined for predetermined times. Sterile disks impregnated with the combined solutions were prepared and placed on Mueller-Hinton plates that were seeded with Pseudomonas or Aspergillus sp. Zones of growth inhibition were measured at postincubation hours 24 and 48. RESULTS: Tobramycin alone inhibited growth of Pseudomonas sp, whereas miconazole inhibited growth of Aspergillus sp. Significant differences in zones of growth inhibition when atropine was combined with tobramycin, when gentamicin was combined with miconazole, or when atropine was combined with miconazole and gentamicin, were not detected. CLINICAL RELEVANCE: Combining selected ophthalmic therapeutic agents for as long as 6 hours does not appear to alter the in vitro efficacy of the agents against microorganisms used in this study.

H3+ in dense and diffuse clouds.

Interstellar H3+ has been detected in dense as well as diffuse clouds using three 3.7 microns infrared spectral lines of the nu 2 fundamental band. Column densities of H3+ from (1.7-5.5) x 10(14) cm-2 have been measured in dense clouds in absorption against the infrared continua of the deeply embedded young stellar objects GL2136, W33A, MonR2 IRS 3, GL961E, and GL2591. Strong and broad H3+ absorptions have been detected in dense and diffuse clouds towards GC IRS 3 and GCS3-2 in the region of the galactic center. A large column density of H3+, comparable to that of a dense cloud, has been detected towards the visible star Cygnus OB2 No. 12, which has a line of sight that crosses mostly diffuse clouds. The H3+ chemistry of dense and diffuse clouds are discussed using a very simple model. Some future projects and problems are discussed.

Detection of H3+ in the diffuse interstellar medium toward Cygnus OB2 No. 12.

The molecular ion H3+ is considered the cornerstone of interstellar chemistry because it initiates the reactions responsible for the production of many larger molecules. Recently discovered in dense molecular clouds, H3+ has now been observed in the diffuse interstellar medium toward Cygnus OB2 No. 12. Analysis of H3+ chemistry suggests that the high H3+ column density (3.8 x 10(14) per square centimeter) is due not to a high H3+ concentration but to a long absorption path. This and other work demonstrate the ubiquity of H3+ and its potential as a probe of the physical and chemical conditions in the interstellar medium.

Water on the sun.

High-resolution infrared spectra of sunspot umbrae have been recorded with the 1-meter Fourier transform spectrometer on Kitt Peak. The spectra contain a very large number of water absorption features originating on the sun. These lines have been assigned to the pure rotation and the vibration-rotation transitions of hot water by comparison with high-temperature laboratory emission spectra.

Administrative tools for operationalizing a continuum of care for children and youth with severe emotional disturbance.

Critical administrative functions in operationalizing a system of care approach to service delivery include: defining and selecting clients to be served, planning and developing responsive services, and ensuring that services are provided in the most therapeutic and least restrictive family and community-based environments. This article presents an overview of several administrative strategies and mechanisms developed by the Continuum of Care for Emotionally Disturbed Children in South Carolina to carry out these functions. Administrative tools are described, as well as the assumptions underlying their development. Evaluation methods, which include analyzing existing client data, surveying parents, and conducting research within a public/academic partnership, are also briefly described.

A literature review: HIV seropositivity in the elderly.

1. AIDS affects all segments of the population, including the elderly. Elders at risk are recipients of blood products prior to 1985 and their spouses; and persons having unprotected anal or vaginal intercourse outside of a mutually monogamous relationship. 2. Because of stereotyping and lack of awareness, health-care professionals have underserved the needs of the elderly for AIDS care. 3. Elderly persons experience the manifestations of HIV in unique ways, and the consequences of the infection are particularly destructive to their lifestyles and support systems. 4. There is an acute need for nursing intervention in the area of AIDS education targeted to the elderly population. In addition, nurses should be at the forefront of development of a comprehensive home health plan geared specifically for the elderly AIDS client.

Detection of C5 in the Circumstellar Shell of IRC+10216.

The C(5) molecule has been identified in the infrared spectrum of the prototypical obscured carbon star, IRC+10216. In addition to their astrophysical importance, pure carbon chain molecules such as C(5) are of interest in the chemistry of flames and propellants.

Detection of C3 in the Circumstellar Shell of IRC+10216.

Vibration-rotation lines of C(3) have been identified in the circumstellar spectrum of the obscured carbon star IRC+10216. This molecule is of interest in both the chemistry of flames, where it may be involved in the formation of soot, and in astrophysics, where it is a potential building block for carbonaceous grains. This high-resolution infrared detection of the pure carbon chain molecule C(3) allows the estimation of the equilibrium C-C bond length, 1.297 angstroms. Possible astrophysical formation and destruction mechanisms for C(3) are reviewed, including the relationship between C(3) and carbon clusters.