On the fate of sinking diatoms: the transport of active buoyancy-regulating cells in the ocean.

Diatoms are one of the most abundant, diverse and ecologically relevant phytoplanktonic group, contributing enormously to global biogeochemical processes like the carbon and silica cycles. This large success has been partly attributed to the mechanical and optical properties of the silica shell (the frustule) that envelops their body. But since they lack motility it is difficult to conceive how they cope with the fast-fluctuating environment they live in and where distributions of resources are very heterogeneous and dynamical. This pinpoints an important but yet poorly understood feature of diatoms physiology: buoyancy regulation that helps them controlling their sinking speed and position in the water column. While buoyancy regulation by light and nutrients availability has been well studied, the effect of hydromechanical stress via fluid shear has been rather overlooked when considering diatoms dynamics. Here, we aim to start filling this gap by first presenting direct experimental evidences for buoyancy control in response to hydro-mechanical stress and then review recent theoretical models where simple couplings between local shear and buoyancy control always result in heterogeneous cell distributions, specific accumulation regions within complex flows and increased sedimentation times to the depths, features of direct ecological relevance. We conclude by suggesting future experiments aiming to unveil such coupling and therefore gain better understanding on the fate of these fascinating microorganisms in their natural habitat. This article is part of the theme issue 'Stokes at 200 (part 2)'.

Onsite advanced biocleaning system for historical wall paintings using new agar-gauze bacteria gel.

AIMS: This study reports the results of the application of a new agar-gauze biogel system activated with viable bacterial cells to altered wall paintings. METHODS AND RESULTS: Biocleaning using agar biogel and agar-gauze biogel systems was performed onsite by direct application to altered wall painting surfaces (25-1000 cm2 ). The treatments were performed for the restoration of two original Italian sites: (i) at the Vatican Museums, Cristo che salva Pietro dalle acque-La Navicella, a wall painting by Giovanni Lanfranco (1627-1628) and (ii) at Pisa Cathedral Cupola, Incarnato, a wall painting by Orazio Riminaldi (1593-1630) and his brother Girolamo Riminaldi. The novelty of this study is the use of viable Pseudomonas stutzeri A29 cells in an advanced agar-gauze biogel system and the short bio-application contact times of between 3 and 12 h. The historical artworks were altered by lipid and protein residues from past restoration, as confirmed by Py-gas chromatography-mass spectrometry and FT-IR data. The effectiveness of the biological treatment was assessed, and general considerations were discussed. CONCLUSIONS: The short bio-application contact time of advanced agar-gauze gel activated with viable P. stutzeri cells makes this biotechnology promising as an alternative method to the traditional onsite cleaning techniques currently in use for altered historical wall paintings. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, we report for the first time the biocleaning of altered materials located in vertical and vaulted areas using agar-gauze biogel with short application times. These findings are of great significance for future restoration activities and are crucial for determining the best preservation strategies in this field.

Hi-tech restoration by two-steps biocleaning process of Triumph of Death fresco at the Camposanto Monumental Cemetery (Pisa, Italy).

AIMS: In this work, the 'hi-tech' complex biocleaning and restoration of the 14th-century fresco Triumph of Death (5·6 × 15·0 m) at the Camposanto Monumental Cemetery (Pisa, Italy) is reported. Since 2000, the restoration based on the biological cleaning of noble medieval frescoes, has been successfully utilized in this site. METHODS AND RESULTS: The novelty of this study is the two-steps biocleaning process using Pseudomonas stutzeri A29 viable cells, previously applied for recovering other valuable frescoes. In this case, after the fresco detachment from the asbestos-cement support (eternity), both the animal glue and the residues of calcium caseinate were biologically removed respectively from the front and from the back of the fresco in 3 h as indicated by GC-MS and PY/GC-MS analyses. The data obtained during the monitoring of the biorestoration process confirmed that the adopted procedure does not leave residual cells on the fresco surfaces as showed by plate count method, ATP determination and also SEM observation. In addition, to avoid the risk of condensation phenomena after the relocation of the restored fresco sections onto the original walls, the use of a new support has been set up together with the design of a control system that allows a continuous monitoring of environmental parameters for prevention and conservation purposes. CONCLUSIONS: This large-scale biorestoration work clearly shows and confirms that this biotechnology is highly efficient, safe, noninvasive, risk-free and very competitive compared to the traditional cleaning methods, offering an unusual 'resurrection' of the degraded artworks also in very complicated and delicate conditions such as the Triumph of Death fresco, defined for its dimension and artistic importance the 'Pisa's Sistina frescoes'. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings can be of significant importance for other future new restoration activities and they are crucial for determining preservation strategies in this field.

Mineralocorticoid receptor associates with pro-inflammatory bias in the hippocampus of spontaneously hypertensive rats.

Damage observed in the hippocampus of the adult spontaneously hypertensive rat (SHR) resembles the neuropathology of mineralocorticoid-induced hypertension, supporting a similar endocrine dysfunction in both entities. In the present study, we tested the hypothesis that increased expression of the hippocampal mineralocorticoid receptor (MR) in SHR animals is associated with a prevalent expression of pro-inflammatory over anti-inflammatory factors. Accordingly, in the hippocampus, we measured mRNA expression and immunoreactivity of the MR and glucocorticoid receptor (GR) using a quantitative polymerase chain reaction and histochemistry. We also measured serum-glucocorticoid-activated kinase 1 (Sgk1 mRNA), the number and phenotype of Iba1+ microglia, as well as mRNA expression levels of the pro-inflammatory factors cyclo-oxygenase 2 (Cox2), Nlrp3 inflammasome and tumour necrosis factor α (Tnfα). Expression of anti-inflammatory transforming growth factor (Tgf)ß mRNA and the NADPH-diaphorase activity of nitric oxide synthase (NOS) were also determined. The results showed that, in the hippocampus of SHR rats, expression of MR and the number of immunoreactive MR/GR co-expressing cells were increased compared to Wistar-Kyoto control animals. Expression of Sgk1, Cox2, Nlrp3 and the number of ramified glia cells positive for Iba1+ were also increased, whereas Tgfß mRNA expression and the NADPH-diaphorase activity of NOS were decreased. We propose that, in the SHR hippocampus, increased MR expression causes a bias towards a pro-inflammatory phenotype characteristic for hypertensive encephalopathy.

Selective Oestrogen Receptor Agonists Rescued Hippocampus Parameters in Male Spontaneously Hypertensive Rats.

Spontaneously hypertensive rats (SHR) show pronounced hippocampus alterations, including low brain-derived neurotrophic factor (BDNF) expression, reduced neurogenesis, astrogliosis and increased aromatase expression. These changes are reverted by treatment with 17ß-oestradiol. To determine which oestradiol receptor (ER) type is involved in these neuroprotective effects, we used agonists of the ERα [propylpyrazole triol (PPT)] and the ERß [diarylpropionitrite (DPN)] given over 2 weeks to 4-month-old male SHR. Wistar Kyoto normotensive rats served as controls. Using immunocytochemistry, we determined glial fibrillary protein (GFAP)+ astrocytes in the CA1, CA3 and hilus of the dentate gyrus of the hippocampus, aromatase immunostaining in the hilus, and doublecortin (DCX)+ neuronal progenitors in the inner granular zone of the dentate gyrus. Brain-derived neurotrophic factor mRNA was also measured in the hippocampus by the quantitative polymerase chain reaction. In SHR, PPT had no effect on blood pressure, decreased astrogliosis, slightly increased BDNF mRNA, had no effect on the number of DCX+ progenitors, and increased aromatase staining. Treatment with DPN decreased blood pressure, decreased astrogliosis, increased BDNF mRNA and DCX+ progenitors, and did not modify aromatase staining. We hypothesise that, although both receptor types may participate in the previously reported beneficial effects of 17ß-oestradiol in SHR, receptor activation with DPN may preferentially facilitate BDNF mRNA expression and neurogenesis. The results of the present study may help in the design of ER-based neuroprotection for the encephalopathy of hypertension.

Presence of pharmaceuticals and heavy metals in the waters of a Mediterranean coastal wetland: Potential interactions and the influence of the environment.

The occurrence of 17 relevant pharmaceuticals and 7 heavy metals in the waters of the Pego-Oliva Marsh Natural Park (Valencia Community, Spain) were monitored. Thirty four zones (including the lagoon and the most important irrigation channels), covering the main land uses and water sources, were selected for sampling. Thirty three of them were contaminated with at least one pharmaceutical. Ibuprofen and codeine were the pharmaceuticals more frequently detected, in concentrations between 4.8 and 1.2 ng/L and a maximum of 59 ng/L and 63 ng/L, respectively. Regarding metals, Zn showed values under the detection limit in all the samples, while Cd, Co, Cr, Cu, Ni and Pb were detected at concentrations lower than the WHO and EU maximum levels for drinking waters. Ni showed significant direct correlations with diazepam, norfloxacin, ofloxacin and fenofibrate, and inverse relationships with ibuprofen, at 99 and 95% of significance. Cu, Co and Cr also showed significant correlations with some of the pharmaceuticals. These interactions could favor the synergistic/antagonistic interactions among pharmaceuticals and metals in the marsh, which can affect its aquatic fauna and flora or even human health. The influences of the water sources, land uses and spatial distribution of both types of contaminants were also studied.

Análisis retrospectivo sobre la extubación en pacientes con patología neurológica aguda / Retrospective analysis on the extubation in patients with acute neurological pathology

Objetivo: Evaluar la idoneidad de la prueba de respiración espontánea para predecir el fracaso de la extubación de pacientes neurológicos y determinar los factores predictivos de fracaso. Diseño: Casos y controles. De enero de 2001 a diciembre de 2010. Ámbito: Unidad de Cuidados Intensivos. Pacientes: Enfermos neurológicos agudos sometidos a ventilación mecánica y posterior extubación. Se excluyeron: pacientes con cirugías neurológicas programadas, con patología neuromuscular, lesión medular, traqueotomía, politraumatismos con predominio de afectación del resto de los sistemas sobre el neurológico, aquellos que murieron en la Unidad de Cuidados Intensivos o que fueron trasladados. Variables de interés: Tasa de fracaso, infección intrahospitalaria, necesidad de traqueotomía, duración de la ventilación mecánica, estancia en la Unidad de Cuidados Intensivos y en el hospital, mortalidad en esta Unidad, en el hospital y a los 90 días, y factores asociados al fracaso. Resultados: De 479 pacientes, 208 fueron sometidos a prueba de respiración espontánea y posterior extubación. Cincuenta y cuatro (26%) fracasaron, la tasa de complicaciones, la estancia, la duración de la ventilación mecánica y la mortalidad fueron mayores que en el grupo de éxito. Los pacientes con accidente cerebrovascular [OR 4,256 (IC95% 1,442-12,561), p = 0,009] y necesidad de aspiraciones frecuentes [OR 5,699 (IC95% 1,863-17,432), p = 0,002] son más propensos al fracaso [ROC 0,73 (IC95% 0,628-0,840)]. Conclusiones: Los pacientes neurológicos presentan una elevada tasa de fracaso de la extubación con numerosas complicaciones asociadas y muerte. La prueba de respiración espontánea no predijo el éxito de la extubación. Los pacientes con accidente cerebrovascular y necesidad de aspiraciones frecuentes de secreciones se verían abocados a un mayor fracaso de extubación.(AU)

Objective: To assess the adequacy of the spontaneous breathing test to predict extubation failure in neurological patients undergoing mechanical ventilation and to determine factors associated with extubation failure. Design: Case-control study. Between January 2001 and December 2010. Setting: Intensive Care Unit. Patients: Acute neurological patients who underwent mechanical ventilation and were subsequently extubated. Patients with scheduled neurosurgery intervention, neuromuscular disease, spinal cord injury, tracheotomy, multiple trauma with less neurological damage than in other systems, those who died in the Intensive Care Unit or in hospital or those transferred to other hospital, were excluded. Variables of interest: Extubation failure rate, nosocomial infection, need for tracheostomy, duration of mechanical ventilation, ICU and hospital stay, mortality in the ICU or hospital, and at day 90, as well as failure-related factors. Results: Two-hundred and eight patients underwent spontaneous breathing trial, and were subsequently extubated. Fifty-four (26%) patients failed. Patients who failed extubation had a higher complication rate, received mechanical ventilation for more days, their hospitalization was longer, and the mortality rate was higher than in the success group. Patients with stroke [OR 4.256 (95%CI, 1.442-12.561), p=0.009] and those who required a greater number of aspirations during weaning [OR 5.699 (95%CI, 1.863-17.432), p=0.002] were susceptible to extubation failure [ROC curve 0.73 (0.628-0.840)]. Conclusion: Extubation failure in neurological patients is common and frequently associated with severe complications. The spontaneous breathing trial does not predict a successful extubation. Patients with stroke and those who need frequent aspiration of secretions would be doomed to further failure of extubation(AU)

Effects of 17ß-estradiol on the cytoarchitecture of pyramidal CA1 neurons in normoglycemic and diabetic male spontaneously hypertensive rats.

Previous work has shown a reduction of apical dendritic length and spine density in neurons from the CA1 hippocampus subfield of spontaneously hypertensive rats (SHRs). These abnormalities are prevented by treatment for 2 weeks with 17ß-estradiol. In view of the fact that diabetes and hypertension are comorbid diseases, we have now studied the effect of Streptozotocin-induced diabetes on the dendritic tree and spines of CA1 hippocampus neurons, and also compared the regulation of these parameters by 17ß-estradiol in diabetic and normoglycemic SHR. Twenty-week-old male SHR received i.v. 40-mg/kg Streptozotocin or vehicle and studied 1 month afterward. A group of normoglycemic and hyperglycemic SHR also received s.c. a single 17ß-estradiol pellet or vehicle for 2weeks. Hippocampus sections were impregnated with silver nitrate following a modified Golgi's method and the arbor of CA1 pyramidal neurons analyzed by Sholl's method. 17ß-Estradiol treatment of normoglycemic SHR reversed the reduced length of apical dendrites, the low spine density and additionally decreased blood pressure (BP). Diabetic SHR showed increased length of apical and basal dendrites but reduced spine density compared to normoglycemic SHR. Diabetes also decreased BP of SHR. Treatment with 17ß-estradiol of diabetic SHR enhanced dendritic length, increased dendritic spine density and further decreased BP. Thus, changes of cytoarchitecture of CA1 neurons due to 17ß-estradiol treatment of normoglycemic SHR persisted after diabetes induction. A decrease of BP may also contribute to the central effects of 17ß-estradiol in SHR diabetic rats.

17α-Oestradiol-induced neuroprotection in the brain of spontaneously hypertensive rats.

17ß-oestradiol is a powerful neuroprotective factor for the brain abnormalities of spontaneously hypertensive rats (SHR). 17α-Oestradiol, a nonfeminising isomer showing low affinity for oestrogen receptors, is also endowed with neuroprotective effects in vivo and in vitro. We therefore investigated whether treatment with 17α-oestradiol prevented pathological changes of the hippocampus and hypothalamus of SHR. We used 20-week-old male SHR with a blood pressure of approximately 170 mmHg receiving s.c. a single 800 µg pellet of 17α-oestradiol dissolved in cholesterol or vehicle only for 2 weeks Normotensive Wistar-Kyoto (WKY) rats were used as controls. 17α-Oestradiol did not modify blood pressure, serum prolactin, 17ß-oestradiol levels or the weight of the testis and pituitary of SHR. In the brain, we analysed steroid effects on hippocampus Ki67+ proliferating cells, doublecortin (DCX) positive neuroblasts, glial fibrillary acidic protein (GFAP)+ astrocyte density, aromatase immunostaining and brain-derived neurotrophic factor (BDNF) mRNA. In the hypothalamus, we determined arginine vasopressin (AVP) mRNA. Treatment of SHR with 17α-oestradiol enhanced the number of Ki67+ in the subgranular zone and DCX+ cells in the inner granule cell layer of the dentate gyrus, increased BDNF mRNA in the CA1 region and gyrus dentatus, decreased GFAP+ astrogliosis in the CA1 subfield, and decreased hypothalamic AVP mRNA. Aromatase expression was unmodified. By contrast to SHR, normotensive WKY rats were unresponsive to 17α-oestradiol. These data indicate a role for 17α-oestradiol as a protective factor for the treatment of hypertensive encephalopathy. Furthermore, 17α-oestradiol is weakly oestrogenic in the periphery and can be used in males.

Progesterone attenuates several hippocampal abnormalities of the Wobbler mouse.

It is now recognised that progesterone plays a protective role for diseases of the central nervous system. In the Wobbler mouse, a model of motoneurone degeneration, progesterone treatment prevents spinal cord neuropathology and clinical progression of the disease. However, neuropathological and functional abnormalities have also been discovered in the brain of Wobbler mice and patients with amyotrophic lateral sclerosis. The present study examined the hippocampus of control and afflicted Wobbler mice and the changes in response to progesterone treatment. Mice received either a single progesterone implant (20 mg for 18 days). We found that the hippocampal pathology of the untreated Wobblers involved a decreased expression of brain-derived neurotrophic factor (BDNF) mRNA, decreased astrogliosis in the stratum lucidum, stratum radiatum and stratum lacunosum-moleculare, decreased doublecortin (DCX)-positive neuroblasts in the subgranular zone of the dentate gyrus and a decreased density of GABA immunoreactive hippocampal interneurones and granule cells of the dentate gyrus. Although progesterone did not change the normal parameters of control mice, it attenuated several hippocampal abnormalities in Wobblers. Thus, progesterone increased hippocampal BDNF mRNA expression, decreased glial fibrillary acidic protein-positive astrocytes and increased the number of GABAergic interneurones and granule cells. The number of DCX expressing neuroblasts and immature neurones remained impaired in both progesterone-treated and untreated Wobblers. In conclusion, progesterone treatment exerted beneficial effects on some aspects of hippocampal neuropathology, suggesting its neuroprotective role in the brain, in agreement with previous data obtained in the spinal cord of Wobbler mice.

Progesterone down-regulates spinal cord inflammatory mediators and increases myelination in experimental autoimmune encephalomyelitis.

In mice with experimental autoimmune encephalomyelitis (EAE) pretreatment with progesterone improves clinical signs and decreases the loss of myelin basic protein (MBP) and proteolipid protein (PLP) measured by immunohistochemistry and in situ hybridization. Presently, we analyzed if progesterone effects in the spinal cord of EAE mice involved the decreased transcription of local inflammatory mediators and the increased transcription of myelin proteins and myelin transcription factors. C57Bl/6 female mice were divided into controls, EAE and EAE receiving progesterone (100mg implant) 7 days before EAE induction. Tissues were collected on day 17 post-immunization. Real time PCR technology demonstrated that progesterone blocked the EAE-induced increase of the proinflammatory mediators tumor necrosis factor alpha (TNFα) and its receptor TNFR1, the microglial marker CD11b and toll-like receptor 4 (TLR4) mRNAs, and increased mRNA expression of PLP and MBP, the myelin transcription factors NKx2.2 and Olig1 and enhanced CC1+oligodendrocyte density respect of untreated EAE mice. Immunocytochemistry demonstrated decreased Iba1+microglial cells. Confocal microscopy demonstrated that TNFα colocalized with glial-fibrillary acidic protein+astrocytes and OX-42+microglial cells. Therefore, progesterone treatment improved the clinical signs of EAE, decreased inflammatory glial reactivity and increased myelination. Data suggest that progesterone neuroprotection involves the modulation of transcriptional events in the spinal cord of EAE mice.

Increased expression of the mineralocorticoid receptor in the brain of spontaneously hypertensive rats.

The mineralocorticoid receptor (MR) has been considered as both neuroprotective and damaging to the function of the central nervous system. MR may be also involved in central regulation of blood pressure. In the present study, we compared the expression of MR and the glucocorticoid receptor (GR) in the hippocampus and hypothalamus of 16-week-old spontaneously hypertensive rats (SHR) and normotensive control Wistar Kyoto (WKY) rats. In the hippocampus, MR expression was studied by in situ hybridization (ISH), quantitative polymerase chain reaction (PCR) and immunohistochemistry, whereas GR expression was analysed using the latter two procedures. Hypertensive animals showed an increased expression of MR mRNA in the whole hippocampus according to qPCR data and also in CA3 by ISH. Immunocytochemical staining for MR of the dorsal hippocampus, however, did not reveal differences between SHR and WKY rats. SHR showed elevated hypothalamic MR mRNA by qPCR, as well as an increased number of MR immunopositive cells in the magnocellular paraventricular region, compared to WKY rats. By contrast, expression levels of GR mRNA or protein in the hippocampus and hypothalamus of SHR were similar to those of WKY rats. Furthermore, we investigated the role of MR in the hypertensive rats by i.c.v. injection of the MR antagonist RU-2831. This compound produced a significant drop in blood pressure for SHR. In conclusion, MR expression is increased in the hippocampus and hypothalamus of SHR. We suggest that pathological MR overdrive may take responsibility for up-regulation of blood pressure and the encephalopathy of hypertension.

Utilización de la ventilación mecánica no invasiva en neumonía grave por virus H1N1 / Noninvasive mechanical ventilation in severe pneumonia due to H1N1 virus

Objetivo: Análisis del empleo de la VMNI en nuestra serie de pacientes ingresados en la unidad de cuidados intensivos (UCI) afectados por nuevo virus de la gripe A (H1N1), en especial aquellos afectados por neumonía con insuficiencia respiratoria aguda (IRA) hipoxémica grave, observándola necesidad de intubación, mejoría clínico-gasométrica, desarrollo de complicaciones, mortalidad, estancia en UCI y hospitalaria. Diseño: Estudio retrospectivo observacional. Ámbito: UCI del Hospital General de Castellón. Pacientes: Pacientes ingresados en la unidad con neumonía primaria o secundaria, con IRA de predominio hipoxémico. Intervenciones: Se empleó CPAP de Boussignac, sistema Helmet y BiPAP Vision. Resultados: De un total de 10 pacientes ingresados con infección por gripe A H1N1, se empleó laVMNI en 7 (70%) pacientes con un fracaso del 28% (una agudización de asma y otra insuficiencia ventilatoria con obstrucción de vía aérea). Dentro del grupo hipoxémico analizado (5 pacientes),la efectividad de la VMNI fue del 100% en cuanto a mejoría gasométrica y clínica, evitando la intubación de todos estos pacientes. Asimismo, no se produjo ninguna muerte tanto en UCI como en el hospital. La duración (mediana) de la ventilación fue de 6 (4-11) días y la estancia en UCI, de 9 (7-11) días. La tasa de complicaciones fue pequeña (una infección de orina). La tolerancia de la VMNI fue aceptable, destacando el ruido producido por la CPAP. No se produjo ningún contagio en el personal sanitario. Conclusiones: A la luz de los resultados, se podría plantear un mayor empleo de la VMNI ante futuras epidemias (AU)

Objective: The use of noninvasive mechanical ventilation was evaluated in our series of patients admitted to our ICU with pneumonia due to influenza A virus H1N1, assessing the need for intubation, arterial blood gases and clinical improvement, the development of complications and ICU and hospital stay. Design: Retrospective and observational study. Setting: ICU of Castellón University General Hospital (Castellón, Spain).Population: Patients admitted to ICU with pneumonia due to influenza A virus H1N1 and acute hypoxemic respiratory failure. Interventions: Boussignac CPAP, Helmet system and BiPAP Vision® were used. Results: Five of 10 patients with pneumonia and hypoxemia were analyzed, showing 100%effectiveness of noninvasive mechanical ventilation in terms of clinical and arterial blood gas improvement, and avoiding intubation in all cases. There were no patient deaths in ICU or in hospital. The duration (median) of ventilation was 6 (4-11) days, with an ICU stay of 9 (7-11)days. The number of complications was low (except for urinary tract infection due to Pseudomon asaeruginosa), and only the noise produced by CPAP was underscored. There were noinfections among the staff .Conclusions: Based on our results, increased use of noninvasive mechanical ventilation in future epidemics could be proposed (AU)

Progesterone attenuates demyelination and microglial reaction in the lysolecithin-injured spinal cord.

Progesterone treatment of mice with experimental autoimmune encephalomyelitis has shown beneficial effects in the spinal cord according to enhanced clinical, myelin and neuronal-related parameters. In the present work, we report progesterone effects in a model of primary demyelination induced by the intraspinal injection of lysophospatidylcholine (LPC). C57Bl6 adult male mice remained steroid-untreated or received a single 100 mg progesterone implant, which increased circulating steroid levels to those of mouse pregnancy. Seven days afterwards mice received a single injection of 1% LPC into the dorsal funiculus of the spinal cord. A week after, anesthetized mice were perfused and paraffin embedded sections of the spinal cord stained for total myelin using Luxol Fast Blue (LFB) histochemistry, for myelin basic protein (MBP) immunohistochemistry and for determination of OX-42+ microglia/macrophages. Cryostat sections were also prepared and stained for oligodendrocyte precursors (NG2+ cells) and mature oligodendrocytes (CC1+ cells). A third batch of spinal cords was prepared for analysis of the microglial marker CD11b mRNA using qPCR. Results showed that progesterone pretreatment of LPC-injected mice decreased by 50% the area of demyelination, evaluated by either LFB staining or MBP immunostaining, increased the density of NG2+ cells and of mature, CC1+ oligodendrocytes and decreased the number of OX-42+ cells, respect of steroid-untreated LPC mice. CD11b mRNA was hyperexpressed in LPC-treated mice, but significantly reduced in LPC-mice receiving progesterone. These results indicated that progesterone antagonized LPC injury, an effect involving (a) increased myelination; (b) stimulation of oligodendrocyte precursors and mature oligodendrocytes, and (c) attenuation of the microglial/macrophage response. Thus, use of a focal demyelination model suggests that progesterone exerts promyelinating and anti-inflammatory effects at the spinal cord level.

[Noninvasive mechanical ventilation in severe pneumonia due to H1N1 virus]. / Utilización de la ventilación mecánica no invasiva en neumonía grave por virus H1N1.

OBJECTIVE: The use of noninvasive mechanical ventilation was evaluated in our series of patients admitted to our ICU with pneumonia due to influenza A virus H1N1, assessing the need for intubation, arterial blood gases and clinical improvement, the development of complications and ICU and hospital stay. DESIGN: Retrospective and observational study. SETTING: ICU of Castellón University General Hospital (Castellón, Spain). POPULATION: Patients admitted to ICU with pneumonia due to influenza A virus H1N1 and acute hypoxemic respiratory failure. INTERVENTIONS: Boussignac CPAP, Helmet system and BiPAP Vision(®) were used. RESULTS: Five of 10 patients with pneumonia and hypoxemia were analyzed, showing 100% effectiveness of noninvasive mechanical ventilation in terms of clinical and arterial blood gas improvement, and avoiding intubation in all cases. There were no patient deaths in ICU or in hospital. The duration (median) of ventilation was 6 (4-11) days, with an ICU stay of 9 (7-11) days. The number of complications was low (except for urinary tract infection due to Pseudomonas aeruginosa), and only the noise produced by CPAP was underscored. There were no infections among the staff. CONCLUSIONS: Based on our results, increased use of noninvasive mechanical ventilation in future epidemics coujld be proposed.

Involvement of brain-derived neurotrophic factor and neurogenesis in oestradiol neuroprotection of the hippocampus of hypertensive rats.

The hippocampus of spontaneously hypertensive rats (SHR) and deoxycorticosterone (DOCA)-salt hypertensive rats shows decreased cell proliferation and astrogliosis as well as a reduced number of hilar cells. These defects are corrected after administration of 17ß-oestradiol (E(2) ) for 2 weeks. The present work investigated whether E(2) treatment of SHR and of hypertensive DOCA-salt male rats modulated the expression of brain-derived neurotrophic factor (BDNF), a neurotrophin involved in hippocampal neurogenesis. The neurogenic response to E(2) was simultaneously determined by counting the number of doublecortin-immunopositive immature neurones in the subgranular zone of the dentate gyrus. Both hypertensive models showed decreased expression of BDNF mRNA in the granular zone of the dentate gyrus, without changes in CA1 or CA3 pyramidal cell layers, decreased BDNF protein levels in whole hippocampal tissue, low density of doublecortin (DCX)-positive immature neurones in the subgranule zone and decreased length of DCX+ neurites in the dentate gyrus. After s.c. implantation of a single E(2) pellet for 2 weeks, BDNF mRNA in the dentate gyrus, BDNF protein in whole hippocampus, DCX immunopositive cells and the length of DCX+ neurites were significantly raised in both SHR and DOCA-salt-treated rats. These results indicate that: (i) low BDNF expression and deficient neurogenesis distinguished the hippocampus of SHR and DOCA-salt hypertensive rats and (ii) E(2) was able to normalise these biologically important functions in the hippocampus of hypertensive animals.

Prominently decreased hippocampal neurogenesis in a spontaneous model of type 1 diabetes, the nonobese diabetic mouse.

In human diabetes, degenerative and functional disorders of the central nervous system, including depression, are common findings. Defective dentate gyrus (DG) neurogenesis is associated with affective-related disorders and depression. We previously demonstrated reduced DG neurogenesis in a pharmacological type 1 diabetes model, the streptozotocin (STZ)-treated mouse. Here, we explored DG neurogenesis in a spontaneous T1D model, the nonobese diabetic (NOD) mouse, at prediabetic and diabetic stages. Cell proliferation was assessed in the DG of 5, 8 and 12-week-old control C57BL/6 and BALB/c strains and NOD mice, killed 2 h after bromodeoxyuridine (BrdU) administration. Survival of the newly generated cells was studied in 15-week-old animals that were killed 21 days after BrdU injection. The number of proliferative BrdU-positive cells in the DG was, regardless of age, constantly and significantly lower in NOD than in control strains, showing the presence of hippocampal alterations far before clinical diabetes onset in NOD mice. Diabetes also strongly decreased cell survival in NOD DG. However, cell phenotype proportion, as assessed by co-localization with neuronal or glial markers and confocal microscopy, was not modified. Hippocampal neurogenesis is strongly diminished in the spontaneous NOD model, like in the STZ model. Notably, NOD hippocampal DG cell proliferation defect takes place during the prediabetic stage. Whether this early alteration might result, in this autoimmune strain, from hypothalamo-pituitary adrenal axis alterations and/or ongoing brain inflammatory process sharing many characteristics of aging is discussed and deserves further investigation.