Functional upregulation of H+-ATPase by lethal acid stress in cultured inner medullary collecting duct cells

The response ofH⁺-ATPase to lethal acid stress isunknown. A mutant strain (called NHE2d) was derived from cultured inner medullary collecting duct cells (mIMCD-3 cells) following three cyclesof lethal acid stress. Cells were grown to confluence on coverslips,loaded with2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein, andmonitored for intracellular pH(pH_i) recovery from an acid load. The rate of Na⁺-independentpH_i recovery from an acid load inmutant cells was approximately fourfold higher than in parent cells(P < 0.001). TheNa⁺-independentH⁺ extrusion was ATP dependent and K⁺ independent and wascompletely inhibited in the presence of diethylstilbestrol, N, N'-dicyclohexylcarbodiimide,or N-ethylmaleimide. Theseresults indicate that theNa⁺-independentH⁺ extrusion in cultured medullarycells is mediated via H⁺-ATPaseand is upregulated in lethal acidosis. Northern hybridization experiments demonstrated that mRNA levels for the 16- and 31-kDa subunits of H⁺-ATPase remainedunchanged in mutant cells compared with parent cells. We propose thatlethal acid stress results in increased H⁺-ATPase activity in innermedullary collecting duct cells. Upregulation ofH⁺-ATPase could play a protectiverole against cell death in severe intracellular acidosis.

     

Experimentation on Degradation of Petroleum in Contaminated Soils in the Root Zone of Maize (Zea Mays L.) Inoculated with Piriformospora Indica

Plant-based methods such as rhizodegradation are very promising for the remediation of petroleum-contaminated soils. Associations of plants with endophytes can further enhance their phytoremediation potential. In this study, a rhizobox experiment was conducted to investigate whether inoculation with the root-colonizing fungus Piriformospora indica could further enhance the degradation of petroleum hydrocarbons in the root zone of maize (Zea mays L.). The rhizoboxes were subdivided into compartments in accordance with distance from the plants. After filling the boxes with soil from a petroleum-contaminated site, seedlings that had either been inoculated with P. indica or not were grown in the middle compartments of the rhizoboxes and grown for 64 days. A plant-free treatment was included for control. The presence of roots strongly increased the counts of total and petroleum-degrading soil bacteria, respiration, dehydrogenase activity, water-soluble phenols and petroleum degradation. All these effects were also found in the soil adjacent to the middle compartments of the rhizoboxes, but strongly decreased further away from it. Inoculation with P. indica further enhanced all the recorded parameters without changing the spatial pattern of the effects. Inoculated plants also produced around 40% more root and shoot biomass than noninoculated plants and had greener leaves. Together, the results indicate that the treatment effects on the recorded soil microbial and biochemical parameters including petroleum hydrocarbon degradation were primarily due to increased root exudation. Irrespectively of this, they show that maize can be used to accelerate the rhizodegradation of petroleum hydrocarbons in soil and that inoculation with P. indica can substantially enhance the phytoremediation performance of maize.     

Reduction of marginal mass required for successful islet transplantation in a diabetic rat model using adipose tissue–derived mesenchymal stromal cells

Background aims

Adipose tissue–derived mesenchymal stromal cells (AT-MSCs), widely known as multipotent progenitors, release several cytokines that support cell survival and repair. There are in vitro and in vivo studies reporting the regenerative role of AT-MSCs possibly mediated by their protective effects on functional islet cells as well as their capacity to differentiate into insulin-producing cells (IPCs).

Tube Formation Potential of BMSCs and USSCs in Response to HIF-1α Overexpression under Hypoxia

Despite numerous studies on therapeutic effects of mesenchymal stem cells on ischemic tissue regeneration, including angiogenesis, their mechanism of action remains ambiguous. Due to the scarce of investigations based on different stem cell sources with known inherent molecular differences, present study compare tube formation of Bone marrow Mesenchymal Stem Cells and Unrestricted Somatic Stem Cells with known reported different Hox gene expression profile in response to HIF-1α overexpression under hypoxia. This might shed light on some parameters for selection of more responsive source with improved therapeutic effects. Superior in vitro tube formation on Matrigel substratum has been observed by Unrestricted Somatic Stem Cells compared to Bone marrow Mesenchymal Stem Cells which might possibly be due to the discriminating molecular properties of stem cell sources. It may help choosing the appropriate stem cell type for a given therapeutic expectations and also suggests some potential targets for future genetic modification of stem cells.     

Enhanced chondrogenic differentiation of dental pulp-derived mesenchymal stem cells in 3D pellet culture system: effect of mimicking hypoxia

High incidence of articular cartilage defects resulting from age-related degeneration or trauma injuries is a major problem worldwide. Limited self-regeneration ability of cartilage often leads to inappropriate biochemistry and structure of healed tissue. Considering Impairments of traditional treatments, cell-based therapies are promising. The rapid ex vivo expansion and chondrogenic differentiation capability make dental pulp stem cells (DPSCs) a favorable cell type for therapeutic application, however strategies in order to efficient cartilage tissue-like production are imperative. In the present study the potential role of hypoxia mimicking agent, cobalt chloride (CoCl2), on chondrogenic differentiation of human DPSCs was surveyed. Cell viability assay used to obtain the optimum dose and exposure time of CoCl2. DPSCs were differentiated in pellet culture system after CoCl2 pretreatment. Chondrogenic differentiation efficiency was evaluated by histological and immunohistological analyses. The results showed that CoCl2 led to increased pellet size, integrity and matrix deposition with organizations more resembled typical cartilage lacuna structure. Furthermore, CoCl2 could improve differentiation by elevated chondrogenic markers, glycosaminoglycans (GAGs) and collagen II expression. CoCl2 pretreatment mitigated hypertrophy, as well, which was reflected in decreased collagen X expression. Alkaline phosphatase (ALP) specific activity did not change significantly by CoCl2 preconditioning. Based on current study hypoxia mimicking agent, CoCl2, could be suggested to promote DPSCs chondrogenic differentiation.     

The Modulatory Effect of Metabotropic Glutamate Receptor Type-1α on Spike-Wave Discharges in WAG/Rij Rats

Modulatory function of metabotropic glutamate type 1 (mGlu1) receptors plays a crucial role in the pathophysiology of some neurological disorders, including schizophrenia and epilepsy. In this study, the expression of mGlu1α receptors in the thalamic nuclei was assessed during development of absence seizures in the WAG/Rij rats, a valid genetic animal model of absence epilepsy. In addition, the effect of pharmacological modulation of mGlu1α receptors in the laterodorsal (LD) nucleus of the thalamus on the characteristic features of bioelectrical brain activities in the WAG/Rij rats was assessed. The expression of mGlu1α receptors in the LD was assessed in four experimental groups of both WAG/Rij and Wistar rats with 2 and 6 months of age. Agonist and antagonist of mGlu1α receptors were infused in LD in the six months old WAG/Rij (epileptic) rats. The protein level of mGlu1α receptors in the thalamus of the 6-month-old WAG/Rij rats was lower than non-epileptic animals. In addition, the distribution of mGlu1α receptors in different thalamic nuclei was lower in the 6-month-old WAG/Rij compared to age-matched Wistar rats. The gene expression of mGlu1α receptor was also significantly lower in 6-month-old WAG/Rij rats in the LD compared to other animal groups. The microinjection of mGlu1α receptors agonist and antagonist in the LD reduced the duration of spike-wave discharges (SWDs) and increased the amplitude and duration of SWDs, respectively, in 6-month-old WAG/Rij rats. The alterations of mGlu1α receptors expression in the thalamus of epileptic WAG/Rij rats as well as its modulatory effects in the generation of SWDs suggest the potential of mGlu1 receptors as a therapeutic target in absence epilepsy.