Spatial Heterogeneity of Cyanobacteria and Diatoms in a Thermally Stratified Canyon‐Shaped Reservoir

Phytoplankton communities in lakes and reservoirs are seldom homogeneously distributed but usually aggregate in patches and gradients. In this study we have combined the use of in vivo spectrofluorometry and acoustic Doppler current profiling to investigate the effect of water movements on the spatial distribution of cyanobacteria and diatoms in a thermally stratified reservoir in SW Spain. The distinctive canyon‐shaped morphometry of the reservoir (El Gergal) favoured the development of a “conveyor belt” pattern of circulation aligned with the long axis of the reservoir. Under non‐regulated conditions, the spatial distribution of phytoplankton was almost entirely dependent on the interactions between advective transport and the buoyancy properties of the different functional groups of phytoplankton. The positively‐buoyant cyanobacteria accumulated near the surface and were then transported downwind by the surface drift currents. In contrast, the negatively‐buoyant diatoms sank in the water column and were transported upwind by the sub‐surface return currents. When deep water was abstracted from the reservoir, these distribution patterns were modified. The results are discussed in relation to the problem of acquiring representative water samples from the reservoir and the application of a simple empirical model to optimize the location of the station used for routine cyanobacteria sampling on the reservoir. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Review. Cognitive and emotional consequences of binge drinking: role of amygdala and prefrontal cortex

Binge drinking is an increasingly recognized problem within the UK. We have studied the relationship of binge drinking to cognitive and emotional functioning in young adults, and have found evidence for increased impulsivity, impairments in spatial working memory and impaired emotional learning. Since in human studies it is difficult to understand whether such behavioural changes pre-date or are a consequence of binge drinking, we have also studied parallel behaviours in a rodent model, in which rats are exposed to intermittent episodes of alcohol consumption and withdrawal. In this model, and in parallel with our findings in human binge drinkers, and alcoholic patients who have undergone multiple episodes of detoxification, we have found evidence for impairments in aversive conditioning as well as increased impulsivity. These behavioural changes are accompanied by facilitated excitatory neurotransmission and reduced plasticity (long-term potentiation (LTP)) in amygdala and hippocampus. The impaired LTP is accompanied by both impaired associative learning and inappropriate generalization of previously learned associations to irrelevant stimuli. We propose that repeated episodes of withdrawal from alcohol induce aberrant neuronal plasticity that results in altered cognitive and emotional competences.     

Accumbal dopamine function in postpartum rats that were raised without their mothers

Postpartum rats that had been previously raised in an artificial rearing (AR) apparatus, without their mothers or siblings during the preweaning period, show altered maternal responses towards their own offspring in adulthood. In mother-reared (MR) rats, nucleus accumbens (NAC) dopamine (DA) responses to pups evoke a robust sustained rise during the postpartum period and following treatment with estrogen/progesterone parturient-like hormones (Afonso et al., 2009). These MR females had siblings that received AR rearing with varying amounts of preweaning tactile stimulation (ARmin; ARmax). The present study examined NACshell DA responses to pup and food stimuli in these AR rats, and statistically compared them to their MR siblings. Microdialysis samples were collected from adult (90 days postnatal) AR females in different parity states (cycling vs. postpartum, Exp. 1), or after ovariectomy with different hormone treatments (sham vs. hormone, Exp. 2. After basal sample collection, pup and then food stimuli were individually presented to the females in the dialysis chamber. As with their MR siblings, basal DA concentrations were lower and pup-evoked DA responses greater in hormonally-primed AR females than in non-primed AR controls. Compared to their postpartum MR sisters (Exp. 1), AR rats had increased basal DA levels, reduced pup related DA elevations, and disrupted maternal behavior. The postpartum AR impairment in pup-evoked DA was reversed by additional pre-weaning tactile stimulation. Exogenous hormones (Exp. 2) eliminated AR impairments on pup-evoked DA responses. Although MR and AR siblings had comparable DA responses to food stimuli, upon reanalyzing MR data it was found that only postpartum dams had DA responses to pups greater than to food. These data suggest that that the hormonally induced suppression of basal DA levels may reflect saliency of pups which was greater in MR than in AR dams. Preweaning tactile stimulation could partially reverse these effects only in naturally cycling or parturient animals.     

MEK抑制剂抑制吗啡依赖及戒断大鼠脊髓神经元NOS的表达

目的：观察鞘内注射丝裂原活化蛋白激酶抑制剂U0126对吗啡依赖及戒断大鼠戒断症状和痛敏行为以及脊髓神经元NOS表达的影响。方法：采用吗啡依赖及戒断模型,分为正常对照组、依赖组、戒断组（戒断1h）、U0126组,分别作行为学评分（n=8）、免疫组织化学（n=6）和免疫印迹检测（n=4）。结果：①鞘内注射U0126可明显减轻吗啡依赖大鼠戒断症状,戒断组戒断症状评分为28．6±4．89,U0126组为22．5±4．09（P〈0．05）;戒断组TEA评分为13．5±2．55,U0126组为10．0±2．76（P〈0．05）;②鞘内注射U0126可明显减少L5节段脊髓背角Fos阳性神经元的数目,U0126组为287±54,低于戒断组（380±71,P〈0．05）;③U0126组nNOS和iNOS阳性神经元的数目分别为180±32、10．8±2．8,均低于戒断组（239±45,16．8±5．1,P〈0．05）,两给药组脊髓NOS蛋白的表达也显著减少。结论：MEK抑制剂能减轻吗啡依赖及戒断大鼠的戒断症状和在脊髓水平抑制NOS的表达,表明ERK可能参与调控NOS的表达。     

Modulating the structure of phenylalanine‐incorporated ascidiacyclamide through fluorination

We designed four fluorinated Phe‐incorporated ascidiacyclamide ([Phe]ASC) analogs, (cyclo(?Xxx1‐oxazoline2‐d ‐Val3‐thiazole4‐Ile5‐oxazoline6‐d ‐Val7‐thiazole8‐)), [(4‐F)Phe]ASC (Xxx1: 4‐fluorophenylalanine), [(3,5‐F₂)Phe]ASC (Xxx1: 3,5‐difluorophenylalanine), [(3,4,5‐F₃)Phe]ASC (Xxx1: 3,4,5‐trifluorophenylalanine) and [(F₅)Phe]ASC (Xxx1: pentafluorophenylalanine), to modulate the π‐electron density of the aromatic ring of the Phe residue. X‐ray diffraction analysis, ¹H NMR and CD spectra all suggested that the interactions between the benzene ring of the Xxx1 residue and the alkyl groups of oxazoline2 contribute to the stability of the folded structure of these analogs. Substituting fluorines for the hydrogens progressively weakened those interactions through reducing the π‐electron density, thereby mediating transformation from the folded to square structure. As a result, [(F₅)Phe]ASC preferred the square form more than the other analogs did. Also contributing to the preference for the square form may be the hindrance of the rotation around the Cα–Cβ bond by the two ortho‐fluoro substituents of [(F₅)Phe]ASC. These findings demonstrate that the structure of ASC can be modulated by using fluorine as an electron‐withdrawing group. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.     

糖尿病大鼠脊髓内高迁移率族蛋白-1参与调节机械性痛敏的机制

目的:观察高迁移率族蛋白-1(high mobility group box-1,HMGB1)在糖尿病大鼠脊髓内的表达变化,探索其参与糖尿病性机械性痛觉过敏的具体机制,进一步阐明糖尿病性痛的机制,为糖尿病疼痛的治疗提供新的思路。方法:(1)36只SD大鼠随机分成6组(n=6),分别为正常大鼠组、糖尿病大鼠对照组、糖尿病7 d组、14 d、21 d和28 d组。通过Real-time PCR法检测各组大鼠脊髓内HMGB1 m RNA的表达情况。(2)24只SD大鼠分成4组(n=6)制作糖尿病大鼠模型,在造模后第28 d鞘内给予生理盐水、HMGB1的中和抗体10、30和100μg,检测糖尿病大鼠模型在各时间点的机械性缩足阈值。(3)30只SD大鼠随机分成5组(n=6),其中4组给予链尿佐菌素制作糖尿病大鼠模型。模型制作28 d后鞘内给予生理盐水、HMGB1的中和抗体10、30和100μg。另一组大鼠腹腔给予生理盐水,作为糖尿病大鼠的对照组。检测各组大鼠脊髓的TNF-α、IL-1β和IL-6 m RNA的表达。结果:(1)糖尿病大鼠模型制作21 d和28 d,脊髓内HMGB1 m RNA的表达显著上调(P0.05)。(2)糖尿病大鼠鞘内给予HMGB1中和抗体30和100μg后,可以在长达24 h的时间内扭转模型大鼠的机械性痛敏(P0.05)。(3)糖尿病大鼠造模28 d后,鞘内给予HMGB1的中和抗体30和100μg可以明显逆转糖尿病大鼠脊髓内的TNF-α、IL-1β和IL-6 m RNA的表达(P0.05)。结论:糖尿病大鼠脊髓内HMGB1显著上调,鞘内给予HMGB1的中和抗体可以通过抑制脊髓内TNF-α等细胞因子的表达而扭转糖尿病大鼠的机械性痛敏。以上结果提示,脊髓HMGB1可能参与了糖尿病机械性痛敏状态的维持过程。我们的研究对脊髓HMGB1参与糖尿病大鼠的疼痛的机制进行初步的探讨,为糖尿病性痛的治疗提供新的思路。     

Use of poly(DL-lactide-ε-caprolactone) membranes and mesenchymal stem cells from the Wharton's jelly of the umbilical cord for promoting nerve regeneration in axonotmesis: In vitro and in vivo analysis

Cellular systems implanted into an injured nerve may produce growth factors or extracellular matrix molecules, modulate the inflammatory process and eventually improve nerve regeneration. In the present study, we evaluated the therapeutic value of human umbilical cord matrix MSCs (HMSCs) on rat sciatic nerve after axonotmesis injury associated to Vivosorb® membrane. During HMSCs expansion and differentiation in neuroglial-like cells, the culture medium was collected at 48, 72 and 96 h for nuclear magnetic resonance (NMR) analysis in order to evaluate the metabolic profile. To correlate the HMSCs ability to differentiate and survival capacity in the presence of the Vivosorb® membrane, the [Ca²⁺]i of undifferentiated HMSCs or neuroglial-differentiated HMSCs was determined by the epifluorescence technique using the Fura-2AM probe. The Vivosorb® membrane proved to be adequate and used as scaffold associated with undifferentiated HMSCs or neuroglial-differentiated HMSCs. In vivo testing was carried out in adult rats where a sciatic nerve axonotmesis injury was treated with undifferentiated HMSCs or neuroglial differentiated HMSCs with or without the Vivosorb® membrane. Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index (SFI), extensor postural thrust (EPT), and withdrawal reflex latency (WRL).     

Exportin 1-independent nuclear export of GAPDH

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme of the glycolytic pathway. Recent studies have demonstrated an additional role in apoptosis: GAPDH is targeted to the nucleus during apoptotic signalling. This nuclear transport has also been observed in serum-depleted cells, but it is reversible in fibroblasts, in contrast to apoptotic-induced transport (Eur J Cell Biol 80 (2001) 419). Here, we analyse the serum depletion-induced transport processes of GAPDH in NIH 3T3 cells. Prolonged serum depletion did not cause cell death, nuclear fragmentation (hoechst staining) or a significant increase in DNA strand-breaks (comet assay). Using cells expressing green fluorescent protein (GFP)-tagged GAPDH allowed us to monitor its intracellular localisation by confocal laser scanning microscopy (CLSM). Treatment of cells with the exportin1 inhibitor leptomycin B (LMB) did not influence cytoplasmic localisation of GFP-GAPDH, indicating that nuclear targeting of GAPDH is not constitutive and may be altered via a serum-dependent regulatory export process. Suprisingly, the export of nuclear GFP-GAPDH after re-addition of serum to starved cells was not prevented by LMB. Thus, nuclear export of GAPDH upon serum depletion is not mediated by exportin1.     

Inhibition of actin polymerization enhances commitment to and execution of apoptosis induced by withdrawal of trophic support

We have previously shown, using jasplakinolide, that stabilization of the actin cytoskeleton enhanced apoptosis induced upon cytokine withdrawal (Posey and Bierer [1999] J. Biol. Chem. 274:4259-4265). It remained possible, however, that a disruption in the regulation of actin dynamics, and not simply F-actin stabilization, was required to affect the transduction of an apoptotic signal. We have now tested the effects of cytochalasin D, a well-characterized agent that promoted actin depolymerization. Actin depolymerization did not affect CD95 (Fas)-induced death of Jurkat T cells in the time course studied but did enhance the commitment to cytokine withdrawal-induced apoptosis of factor-dependent cell lines. The induction of cell death was not the result of direct cytoskeletal collapse, since treatment of the cells with cytochalasin D in the presence of IL-2 did not promote death. As with jasplakinolide, the enhancement of commitment to apoptosis could be delayed by overexpression of the anti-apoptotic protein Bcl-x(L), but, unlike jasplakinolide, cytochalasin D modestly affected the "execution" stage of apoptosis as well. Taken together, these results suggest that changes in actin dynamics, i.e., the rate of actin polymerization and depolymerization, modulate the transduction of the apoptotic signal committing lymphocytes, withdrawn from required growth factors, to the death pathway.