Effect of Digestion with Phospholipase A2 on Endogenous Protein Phosphorylation in Particulate Fractions from Rat Brain Synaptosomes

The endogenous phosphorylation of synapsin 1 in cyclic AMP-containing media was greatly decreased by digestion of synaptic vesicles and synaptosomal membranes with phospholipase A2, suggesting that the system is functionally dependent on the membrane structure. Treatment of the synaptic vesicle fraction with phospholipase A2 also caused a small but significant inhibition of the Ca2+/calmodulin-dependent phosphorylation of the same protein. The Ca2+/calmodulin-dependent phosphorylation of other major acceptors, and the basal phosphorylation of a 52-kD acceptor enriched in the vesicle fraction, remained unchanged after cleavage of the membrane phospholipids with phospholipase A2. The significance of the selective effect of phospholipase A2 treatment on endogenous membrane phosphorylation is discussed.     

Expression of several growth factors and their receptor genes in human colon carcinomas

We have examined the expression of mRNAs for epidermal growth factor (EGF), transforming growth factor-alpha (TGF-α), EGF receptor (EGFR), PDGF-A chain (PDGFA), PDGF-B chain (PDGFB) and PDGF receptor (PDGFR) genes in seven human colorectal carcinoma cell lines and 18 human colorectal carcinomas. In surgically resected specimens of the 18 colorectal tumors, TGF-α, EGFR, PDGFA, PDGFB and PDGFR mRNAs were detected at various levels in 15 (83%), 9 (50%), 18 (100%), 8 (44%) and 12 (67%), respectively. They were also detected in normal tissues. Interestingly, EGF mRNA was detected in only five (28%) of the tumors, but not in normal mucosa. Expression of EGF was also confirmed immunohistochemically in tumor cells. Of the five tumors expressing EGF, four expressed EGFR mRNA and showed a tendency to invade veins and lymphatics. All the colorectal carcinoma cell lines expressed TGF-α mRNA, and five cell lines expressed EGFR mRNA simultaneously. Production of TGF-α protein by DLD-1 and CoLo320DM cells was confirmed by TGF-α specific monoclonal antibody binding assay. The spontaneous³H-thymidine uptake by DLD-1 was suppressed by an anti-TGF-α monoclonal antibody. PDGFA and PDGFB mRNA were also expressed in four cell lines, but PDGFR and EGF mRNA was not detected. These results suggest that human colorectal carcinomas express multi-loops of growth factors and that TGF-α produced by tumor cells functions as an autocrine growth factor in human colonic carcinoma.     

Acceptors for cyclic AMP-dependent and calcium ion-dependent protein kinases in rat brain cytosol fractions: A comparison of occluded (synaptosomal) cytosol with non-occluded cytosol

Endogenous protein phosphorylation patterns were compared in occluded and non-occluded cytosol fractions prepared from rat forebrain. The occluded fraction was taken as representative of synaptosomal cytosol. One- and two-dimensional autoradiographs revealed the presence in non-occluded cytosol of a substrate for cAMP- and Ca²⁺/calmodulin-dependent protein kinase activities of M_r 300kD, corresponding to phosphorylated microtubule-associated protein-2 (MAP-2); this protein was absent in occluded cytosol. In contrast, a major substrate for protein kinase C was observed exclusively in occluded cytosol after phosphorylation under basal conditions. However, after phosphorylation in the presence of exogenous lipids, approximately equal amounts of the 82kD substrate were detected in both fractions, suggesting that protein kinase C in the occluded fraction was present in a partially activated state. Other minor differences in phosphorylation patterns between the two fractions were observed.Special Issue dedicated to Prof. Eduardo De Robertis.     

Enzymic changes in rabbit and rat mammary gland during the lactation cycle

1. Activities of glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), isocitrate dehydrogenase (EC 1.1.1.42), malate dehydrogenase (EC 1.1.1.37), malate dehydrogenase (decarboxylating) (EC 1.1.1.40), and pyruvate carboxylase (EC 6.4.1.1) were determined in subcellular fractions of mammary gland from rabbits during pregnancy, at different stages of lactation and during weaning. The results were compared with those obtained in similar experiments with rat mammary gland. 2. Three bases of expression of the activity of enzymes in the particle-free supernatant fraction of mammary gland were compared. During lactation, activity expressed per mg. of particle-free supernatant protein (uncorrected for milk protein) correlated well with that expressed per mug. of DNA phosphorus. The disadvantages of expressing activities per g. wet wt. are discussed. 3. The major differences between the two tissues were: (a) neither malate dehydrogenase (decarboxylating) nor a soluble form of pyruvate carboxylase could be detected in rabbit mammary gland at any stage of the lactation cycle; (b) isocitrate dehydrogenase increased in activity during lactation in rabbit mammary gland, but not in that of the rat. 4. Pyruvate carboxylase in the mitochondrial fraction of rabbit mammary gland, and in both the mitochondrial and the soluble fractions of rat mammary gland, did not change in activity during lactation. 5. For each tissue, the NADP-dependent dehydrogenases studied had a high activity at all stages of the lactation cycle compared with the rate of fatty acid synthesis at mid-lactation. The significance of these results is discussed with respect to the supply of NADPH via NADH.     

Oxidative brain and cerebellum injury in diabetes and prostate cancer model: Protective effect of metformin

The body can host the spread of prostate cancer cells. Metastases from prostate cancer are more frequently seen in the brain, liver, lungs, and lymph nodes. A well-known antidiabetic drug, metformin, is also known to have antitumor effects. Our study focuses on the evaluation of potential metformin protective effects on brain and cerebellum damage in streptozotocin (STZ)-induced diabetic and Dunning prostate cancer models. In this investigation, six groups of male Copenhagen rats were created: control, diabetic (D), cancer (C), diabetic + cancer (DC), cancer + metformin, and diabetic + cancer + metformin. The brain and cerebellum tissues of the rats were taken after sacrifice. Oxidative stress markers including reduced glutathione level, lipid peroxidation, glutathione reductase, glutathione peroxidase, glutathione-S-transferase, catalase, superoxide dismutase activities, reactive oxygen species, total oxidant and total antioxidant status, lactate dehydrogenase, xanthine oxidase, acetylcholinesterase activities, protein carbonyl contents, nitric oxide and OH-proline levels, sodium potassium ATPase, carbonic anhydrase, and glucose-6-phosphate dehydrogenase activities; glycoprotein levels including hexose, hexosamine, fucose, and sialic acid levels; and histone deacetylase activity as a cancer marker were determined. Oxidative stress markers were impaired and glycoprotein levels and histone deacetylase activity were increased in the D, C, and DC groups. Metformin therapy reversed these effects. Metformin was found to protect the brain and cerebellum of STZ-induced diabetic rats with Dunning prostate cancer from harm caused by MAT-Lylu metastatic cells.     

Aluminum Toxicity: A Case Study on Tobacco (<i>Nicotiana tabacum</i> L.)

Aluminum is an abundant metal in the earth’s crust that turns out to be toxic in acidic environments. Many plants are affected by the presence of aluminum at the whole plant level, at the organ level, and at the cellular level. Tobacco as a cash crop (Nicotiana tabacum L.) is a widely cultivated plant worldwide and is also a good model organism for research. Although there are many articles on Al-phytotoxicity in the literature, reviews on a single species that are economically and scientifically important are limited. In this article, we not only provide the biology associated with tobacco Al-toxicity, but also some essential information regarding the effects of this metal on other plant species (even animals). This review provides information on aluminum localization and uptake process by different staining techniques, as well as the effects of its toxicity at different compartment levels and the physiological consequences derived from them. In addition, molecular studies in recent years have reported specific responses to Al toxicity, such as overexpression of various protective proteins. Besides, this review discusses data on various organelle-based responses, cell death, and other mechanisms, data on tobacco plants and other kingdoms relevant to these studies.     

Evaluation of scanning resolution,detector choice and detector orientation to be used for accurate and time-efficient commissioning of a 6MV clinical linear accelerator

The present study is aimed at exploring different scanning parameters, detectors and their orientations for time-efficient and accurate commissioning of a 6 MV clinical linear accelerator (LINAC). Beam profiles and percentage depth dose (PDD) curves were measured with a PTW dosimetry diode, a PTW Semiflex and a PinPoint ion chamber in different orientations. To acquire beam data, equidistant (step size of 0.5 mm, 1 mm, 2 mm and 3 mm) and fanline (step size of 2–0.5 mm, 2–1 mm, 3–0.5 mm and 3-1 mm) scanning modes were employed and data measurement time was recorded. Scan time per measurement point was also varied (0.2 s, 0.5 s and 1.0 s) to investigate its effect on the accuracy and acquisition time of beam data. Accuracy of the measured data was analyzed on the basis of the variation between measured data and data modeled by a treatment planning system. Beam profiles (particularly in penumbra region) were found to be sensitive to variation in scanning resolution and showed an improved accuracy with decrease in step size, while PDD curves were affected negligibly. The accuracy of beam data obtained with the PTW dosimetry diode and the PinPoint ion chamber was higher than those obtained with the PTW Semiflex ion chamber for small fields (2?×?2 cm² and 3?×?3 cm²). However, the response of the PTW diode and the PinPoint ion chamber was significantly indifferent in these fields. Furthermore, axial orientation of the PTW Semiflex ion chamber improved accuracy of profiles and PDDs as compared to radial orientation, while such a difference was not significant for the PinPoint ion chamber. It is concluded that a scan time of 0.2 s/point with a fanline scanning resolution of 2–1 mm for beam profiles and 3 mm for PDDs are most favorable in terms of accuracy and time efficiency. For small fields (2?×?2 cm² and 3?×?3 cm²), a PinPoint ion chamber in radial orientation or a dosimetry diode in axial orientation are recommended for both beam profiles and PDDs. If a PinPoint ion chamber and a PTW dosimetry diode are not available, a Semiflex ion chamber in axial orientation may be used for small fields.

     

Facile Synthesis of Hybrid Nanoflowers Using Glycine and Phenylalanine and Investigation of Their Catalytic Activity

In the context of the proposed work, two different amino acids (Glycine, Phenylalanine) have interacted with copper ions in a phosphate buffer (PBS) in place of enzymes. This interaction resulted in the nucleation of copper phosphate crystals and the formation of flower-shaped amino acid-copper hybrid nanostructures (AA-hNFs), which grew through self-assembly. While Cu (II) ions in the structure of AA-hNFs were used as Fenton's agent for the catalytic activity. SEM, energy dispersive X-ray spectroscopy, X-ray diffraction, and Fourier transform infrared spectroscopy measurements were used to define the AA-hNFs′ characterisation. The peroxidase-like activities of AA-hNFs were investigated by UV/VIS spectrophotometer. Metal nanoparticles have peroxidase-like activity. A class of enzymes known as peroxidases is able to catalyze the conversion of hydrogen peroxide into hydroxyl radicals. These radicals also take part in electron transfers with substrates, which results in color during oxidation. When cupric oxide nanoparticles are added to the peroxidase substrate while H₂O₂ is present, a blue color product with a maximum absorbance at=652 nm can result, demonstrating the catalytic activity of a peroxidase. The morphology and composition of AA-hNFs were carefully characterized and the synthesized parameters were optimized systematically. Results showed that the nanoparticles were dispersed with an average diameter of 7–9 μm and indicated a uniform flower shape. The results of the investigation are anticipated to significantly advance a number of technical and scientific sectors.     

Suppression of TLR4/MyD88/TAK1/NF-κB/COX-2 Signaling Pathway in the Central Nervous System by Bexarotene,a Selective RXR Agonist,Prevents Hyperalgesia in the Lipopolysaccharide-Induced Pain Mouse Model

Neurochemical Research - A selective RXR agonist, bexarotene, has been shown to have anti-inflammatory, anti-nociceptive, and neuroprotective effects in several models of numerous neurological...