A hydroxypyrone-based inhibitor of metalloproteinase-12 displays neuroprotective properties in both status epilepticus and optic nerve crush animal models

Recently, we showed that matrix metalloproteinase-12 (MMP-12) is highly expressed in microglia and myeloid infiltrates, which are presumably involved in blood\u2013brain barrier (BBB) leakage and subsequent neuronal cell death that follows status epilepticus (SE). Here, we assessed the effects of a hydroxypyrone-based inhibitor selective for MMP-12 in the pilocarpine-induced SE rat model to determine hippocampal cell survival. In the hippocampus of rats treated with pilocarpine, intra-hippocampal injections of the MMP-12 inhibitor protected Cornu Ammonis 3 (CA3) and hilus of dentate gyrus neurons against cell death and limited the development of the ischemic-like lesion that typically develops in the CA3 stratum lacunosum-moleculare of the hippocampus. Furthermore, we showed that MMP-12 inhibition limited immunoglobulin G and albumin extravasation after SE, suggesting a reduction in BBB leakage. Finally, to rule out any possible involvement of seizure modulation in the neuroprotective effects of MMP-12 inhibition, neuroprotection was also observed in the retina of treated animals after optic nerve crush. Overall, these results support the hypothesis that MMP-12 inhibition can directly counteract neuronal cell death and that the specific hydroxypyrone-based inhibitor used in this study could be a potential therapeutic agent against neurological diseases/disorders characterized by an important inflammatory response and/or neuronal cell loss

A computerized analysis of the entire retinal ganglion cell population and its spatial distribution in adult rats

AbstractIn adult albino (SD) and pigmented (PVG) rats the entire population of retinal ganglion cells (RGCs) was quantified and their spatial distribution analyzed using a computerized technique. RGCs were back-labelled from the optic nerves (ON) or the superior colliculi (SCi) with Fluorogold (FG). Numbers of RGCs labelled from the ON [SD: 82,818±3,949, n=27; PVG: 89,241±3,576, n=6) were comparable to those labelled from the SCi [SD: 81,486±4,340, n=37; PVG: 87,229±3,199; n=59]. Detailed methodology to provide cell density information at small scales demonstrated the presence of a horizontal region in the dorsal retina with highest densities, resembling a visual streak

Characterizing microglia activation: a spatial statistics approach to maximize information extraction

Microglia play an important role in the pathology of CNS disorders, however, there remains significant uncertainty about the neuroprotective/degenerative role of these cells due to a lack of techniques to adequately assess their complex behaviour in response to injury. Advancing microscopy techniques, transgenic lines and well-characterized molecular markers, have made histological assessment of microglia populations more accessible. However, there is a distinct lack of tools to adequately extract information from these images to fully characterise microglia behaviour. This, combined with growing economic pressures and the ethical need to minimise the use of laboratory animals, led us to develop tools to maximise the amount of information obtained. This study describes a novel approach, combining image analysis with spatial statistical techniques. In addition to monitoring morphological parameters and global changes in microglia density, nearest neighbour distance, and regularity index, we used cluster analyses based on changes in soma size and roundness to yield novel insights into the behaviour of different microglia phenotypes in a murine optic nerve injury model. These methods should be considered a generic tool to quantitatively assess microglia activation, to profile phenotypic changes into microglia subpopulations, and to map spatial distributions in virtually every CNS region and disease state

ANTIMICROBIAL ACTIVITY OF MEDICINAL PLANTS FROM THE HUAUTLA SIERRA BIOSPHERE RESERVE IN MORELOS (MÉXICO)

Twenty-two extracts from nine Mexicanmedicinal plants of eight different familiesused for people neighbor to Huautla SierraBiosphere Reserve (REBIOSH) in differentinfectious diseases were assayed in vitro todetermine their antibacterial activity againstStaphylococcus aureus, Streptococcus faecalis; Escherichia coli; Proteus mirabilis;Salmonella typhi and the yeast Candidaalbicans. Most plants showed antibacterialactivity, while two plants showed activityagainst range 0.25 a 4 mg/mL almost onemicroorganism used. The extracts showedminimum inhibitory concentration (MIC)against S. aureus and S. faecalis, representsmicroorganisms Gram-positive were hexanic and acetonic extracts of Bursera copallifera and hexanic extract of B. grandifolia.The ethanol extract of Lippia graveolensshowed a MIC value of 1 mg/mL againstyeast Candida albicans.Se determinó la actividad antimicrobianade veintidós extractos orgánicos de nueveespecies vegetales medicinales de ochodiferentes familias, que son comúnmenteempleadas por los habitantes de las comunidades vecinas a la reserva de la biosferaSierra de Huautla (REBIOSH), Morelos,para tratar diferentes padecimientos infecciosos. Los microorganismos empleadosfueron: Staphylococcus aureus; Streptococcus faecalis; Escherichia coli; Proteusmirabilis; Salmonella typhi y la levaduraCandida albicans. La mayoría de los extractos de las plantas probadas inhibieronel crecimiento microbiano en un rango de0.25 a 4 mg/mL de al menos uno de losmicroorganismos utilizados. Los extractosque mostraron las menores concentracionesmínimas inhibitorias (CMI) frente a losmicroorganismos S. aureus y S. faecalis,representativos de Gram-positivos fueron el extracto hexánico y acetónico de Burseracopallifera y el hexánico de B. grandifolia.El extracto etanólico de Lippia graveolenslogró inhibir a C. albicans con una CMIde 1 mg/mL

Tracing the retina to analyze the integrity and phagocytic capacity of the retinal pigment epithelium

We have developed a new technique to study the integrity, morphology and functionality of the retinal neurons and the retinal pigment epithelium (RPE). Young and old control albino (Sprague-Dawley) and pigmented (Piebald Virol Glaxo) rats, and dystrophic albino (P23H-1) and pigmented (Royal College of Surgeons) rats received a single intravitreal injection of 3% Fluorogold (FG) and their retinas were analyzed from 5 minutes to 30 days later. Retinas were imaged in vivo with SD-OCT and ex vivo in flat-mounts and in cross-sections. Fifteen minutes and 24 hours after intravitreal administration of FG retinal neurons and the RPE, but no glial cells, were labeled with FG-filled vesicles. The tracer reached the RPE 15 minutes after FG administration, and this labeling remained up to 30 days. Tracing for 15 minutes or 24 hours did not cause oxidative stress. Intraretinal tracing delineated the pathological retinal remodelling occurring in the dystrophic strains. The RPE of the P23H-1 strain was highly altered in aged animals, while the RPE of the RCS strain, which is unable to phagocytose, did not accumulate the tracer even at young ages when the retinal neural circuit is still preserved. In both dystrophic strains, the RPE cells were pleomorphic and polymegathic

Etiology and susceptbility to antimicrobials of urinary tract infections in patients of Rehabilitation Unit of a regional hospital

Objetivos: Conocer los resultados del estudio microbiológico de las muestras de orina de pacientes de la Unidad de Rehabilitación de un hospital regional, incluyendo los datos locales de sensibilidad antibiótica. Material y métodos: Se analizó la base de datos del Laboratorio de Microbiología del Hospital Universitario Virgen de las Nieves con los estudios microbiológicos de las muestras de orinas realizados durante el año 2012. Se investigó la etiología y la sensibilidad a los antibióticos de los uropatógenos más frecuentes presentes en 143 muestras de orina obtenidas mediante sondaje o cateterismo. Los resultados del estudio de 9066 muestras de orina de micción media de sujetos estudiados en Atención Primaria fueron utilizados como grupo control. Resultados: El 50% de los pacientes neurológicos presentaron urocultivos positivos, frente al 23% de los controles. E. coli fue el microorganismo más frecuentemente aislado en el grupo de enfermos neurológicos, al igual que en los controles. En el estudio de resistencias E. coli presentó altas tasas de resistencia a ampicilina, cotrimoxazol, ciprofloxacino y amoxicilina con ácido clavulánico. Conclusiones: E. coli es el microorganismo más frecuentemente aislado en los dos grupos de pacientes estudiados. Las resistencias a los antibióticos de los microorganismos procedentes de los dos grupos tienen patrones diferentes, siendo globalmente menor a fosfomicina.Objectives: Knowing the results of microbiological tests of urine samples of patients from the Rehabilitation Unit of a regional hospital, including local data of antibiotic susceptibility. Material and methods: We analyzed the database of the Microbiology Laboratory of the Hospital Universitario Virgen de las Nieves with microbiological studies conducted urine samples during 2012. We investigated the etiology and antimicrobial susceptibility of uropathogens frequently present in 143 urine samples. The 9066 survey results voiding urine mean in Primary Care study subjects were used as controls. Results: The 50% of neurological patients had positive urine cultures, versus 23% of controls. E. coli was the most frequently isolated organism group neurological patients, as in the controls. Resistance studies in E. coli showed high rates of resistance to ampicillin, cotrimoxazole, ciprofloxacin and amoxicillin with clavulanic acid. Conclusions: E. coli is the most frequently isolated microorganism in both groups of patients studied. The antibiotic resistance of microorganisms from the two groups have different patterns, being globally less to fosfomycin

The dark halo of the Hydra I galaxy cluster: core, cusp, cosmological? Dynamics of NGC 3311 and its globular cluster system

NGC 3311 is the central cD galaxy of the Hydra I cluster. We use globular clusters around NGC 3311, combined with kinematical data of the galaxy itself, to investigate the dark matter distribution in the central region of Hydra I. Radial velocities of 118 bright globular clusters, based on VLT/VIMOS mask spectroscopy, are used to calculate velocity dispersions which are well defined out to 100 kpc. NGC 3311 is the most distant galaxy for which this kind of study has been performed. We also determine velocity dispersions of the stellar component from long slit spectroscopy out to 20 kpc. Moreover, we present a new photometric model for NGC 3311 in the V-band. We search for a dark halo which in the context of a spherical Jeans model. We also compare the radial velocity distributions of globular clusters and planetary nebulae. The projected stellar velocity dispersion rises from 185 km/s to 350 km/s at a radius of 20 kpc. The globular cluster dispersion rises as well from 500 km/s at 10 kpc to about 800 km/s at 100 kpc, comparable to the velocity dispersion of the cluster galaxies. A dark matter halo with a core reproduces well the velocity dispersions of stars and globular clusters simultaneously under isotropy. The central stellar velocity dispersions predicted by cosmological NFW halos are less good representations, while the globular clusters allow a wide range of halo parameters. A suspected radial anisotropy of the stellar population aggravates the deviations. However, we find discrepancies with previous kinematical data, which we cannot resolve and may indicate a more complicated velocity pattern. Although one cannot conclusively demonstrate that the dark matter halo of NGC 3311 has a core rather than a cusp, a core seems to be preferred by the present data. A more complete velocity field and an analysis of the anisotropy is required to reach firm conclusions.Comment: 8 pages, 5 figures, abstract abridged, accepted for publication in A&

A gliclazide complex based on palladium towards Alzheimer's disease: Promising protective activity against Aβ-induced toxicity in: C. elegans

A new palladium coordination compound based on gliclazide with the chemical formula [Pd(glz)2] (where glz = gliclazide) has been synthesized and characterised. The structural characterization reveals that this material consists of mononuclear units formed by a Pd2+ ion coordinated to two molecules of the glz ligand, in which palladium ions exhibit a distorted plane-square coordination sphere. This novel material behaves like a good and selective inhibitor of butyrylcholinesterase, one of the most relevant therapeutic targets against Alzheimer's disease. Analysis of the enzyme kinetics showed a mixed mode of inhibition, the title compound being capable of interacting with both the free enzyme and the enzyme-substrate complex. Finally, the palladium compound shows promising protective activity against Aβ-induced toxicity in the Caenorhabditis elegans model, which has never been reported.he authors gratefully acknowledge funding support from the Spanish Ministry of Science, Innovation and Universities (PGC2018-102052-B-C21 and PID2020-116460RB-I00) and the Junta de Andalućıa (FQM-394 and FQM-134). The authors gratefully acknowledge the funding support of FEDER/Junta de Andalućıa Consejería de Economía y Conocimiento, Grant B-AGR-193-UGR18. O. L. and J. G. F. B. also thank Grant PID2020-116460RB-I00 funded by MCIN/AEI/10.13039/50110 0011033. S. R. acknowledge the Juan de la Cierva Fellowship (IJC2019-038894-I)

Comparative evaluation of methods for estimating retinal ganglion cell loss in retinal sections and wholemounts

To investigate the reliability of different methods of quantifying retinal ganglion cells (RGCs) in rat retinal sections and wholemounts from eyes with either intact optic nerves or those axotomised after optic nerve crush (ONC). Adult rats received a unilateral ONC and after 21 days the numbers of Brn3a+ , bIII-tubulin+ and Islet-1+ RGCs were quantified in either retinal radial sections or wholemounts in which FluoroGold (FG) was injected 48 h before harvesting. Phenotypic antibody markers were used to distinguish RGCs from astrocytes, macrophages/microglia and amacrine cells. In wholemounted retinae, counts of FG+ and Brn3a+ RGCs were of similar magnitude in eyes with intact optic nerves and were similarly reduced after ONC. Larger differences in RGC number were detected between intact and ONC groups when images were taken closer to the optic nerve head. In radial sections, Brn3a did not stain astrocytes, macrophages/microglia or amacrine cells, whereas βIII-tubulin and Islet-1 did localize to amacrine cells as well as RGCs. The numbers of βIII-tubulin+ RGCs was greater than Brn3a+ RGCs, both in retinae from eyes with intact optic nerves and eyes 21 days after ONC. Islet-1 staining also overestimated the number of RGCs compared to Brn3a, but only after ONC. Estimates of RGC loss were similar in Brn3astained radial retinal sections compared to both Brn3a-stained wholemounts and retinal wholemounts in which RGCs were backfilled with FG, with sections having the added advantage of reducing experimental animal usage

Chromenopyrazole, a Versatile Cannabinoid Scaffold with in Vivo Activity in a Model of Multiple Sclerosis

A combination of molecular modeling and structure activity relationship studies has been used to fine-tune CB2 selectivity in the chromenopyrazole ring, a versatile CB1/CB2 cannabinoid scaffold. Thus, a series of 36 new derivatives covering a wide range of structural diversity has been synthesized, and docking studies have been performed for some of them. Biological evaluation of the new compounds includes, among others, cannabinoid binding assays, functional studies, and surface plasmon resonance measurements. The most promising compound [43 (PM226)], a selective and potent CB2 agonist isoxazole derivative, was tested in the acute phase of Theiler's murine encephalomyelitis virus-induced demyelinating disease (TMEV-IDD), a well established animal model of primary progressive multiple sclerosis. Compound 43 dampened neuroinflammation by reducing microglial activation in the TMEV