Oxidant, mitochondria and calcium: an overview

Mitochondria are active in the continuous generation of reactive oxygen species (ROS), (e.g., superoxide), thereby favouring a situation of mitochondrial oxidative stress. Under oxidative stress--for example, ischaemia-reoxygenation injury to cells--mitochondria form superoxide, which in turn is converted to hydrogen peroxide and the potent reactive species, hydroxyl radical. Alternatively, mitochondrial superoxide may react with nitric oxide to form potent oxidant peroxynitrite and as a consequence, mitochondrial function is altered. An increase in the release of calcium from mitochondria by oxidants stimulates calcium-dependent enzymes such as calcium-dependent proteases, nucleases, and phospholipases, which subsequently trigger apoptosis of the cells. In principle, calcium can leave mitochondria by different ways: by non-specific leakage through the inner membrane by "pore formation," by changes in the membrane lipid phase, by reversal of the uniport influx carrier, by the specific calcium/hydrogen (or sodium) antiport system, by channel-mediated release pathways, or by a combination of two or more of these pathways. Additionally, the release of calcium from mitochondria can also occur either by oxidation of internal nicotinamide adenine nucleotides to ADP ribose and nicotinamide or by oxidation of thiols in membrane proteins. Once calcium efflux has been triggered, a series of common pathways of apoptosis are initiated, each of which may be sufficient to destroy the cell. Apoptosis requires the active participation of cellular components, and several genes have been suggested to control apoptosis. The proto-oncogene bcl-2 suppresses apoptosis through mitochondrial effects. Overexpression of bcl-2 in the mitochondrial membrane inhibits calcium efflux, but the underlying mechanisms are not clearly known. Further studies are needed to explore the nature of the apoptosis-inducing pathways, the precise mechanisms of calcium efflux, the molecular partners of bcl-2 oncoproteins at the level of the outer-inner membrane contact sites, the molecular biology of the apoptosis-inducing factor formation and release, and the essential molecular targets of apoptosis-inducing proteases. Clarification of these issues might facilitate the understanding of mitochondrial response on cellular calcium dynamics under oxidant stress.     

Plant Growth-Promoting Rhizobacteria-Mediated Adaptive Responses of Plants Under Salinity Stress

In this recent era, several approaches have been developed to alleviate the adverse effects of salinity stress in different plants. However, some of them are not eco-friendly. In this context, evolving sustainable approaches which enhance the productivity of saline soil without harming the environment are necessary. Many recent studies showed that plant growth-promoting rhizobacteria (PGPR) are known to confer salinity tolerance to plants. Salt-stressed plants inoculated with PGPR enhance the growth and productivity of crops by reducing oxidative damage, maintaining ionic homeostasis, enhancing antioxidant machinery, and regulating gene expressions. The PGPR also regulates the photosynthetic attributes such as net photosynthetic rate, chlorophyll, and carotenoid contents and enhances the salinity tolerance to plants. Moreover, PGPR has a great role in the enhancement of phytohormones and secondary metabolites synthesis in plants under salt stress. This review summarizes the current reports of the application of PGPR in plants under salt stress and discusses the PGPR-mediated mechanisms in plants of salt tolerance. This review also discusses the potential role of PGPR in cross-talk with phytohormones and secondary metabolites to alleviate salt stress and highlights the research gaps where further research is needed.

     

Enhancement of methionine production by methionine analogue ethionine resistant mutants of Brevibacterium heali

In order to improve the methionine yield of the isolate B. heali, attempts were made to isolate mutants resistant to the methionine analogue DL-ethionine after mutagenesis with N-methyl-N′-nitro-N-nitrosoguanidine (NTG). The minimum inhibitory concentration (MIC) of ethionine for B. heali was found to be 2 mM. After mutagenesis and screening, five mutants resistant to 50 mM of ethionine were isolated. The yield of the best ethionine resistant mutant, B. heali Br EthR, was 13 mg/l of methionine medium under optimum cultivation conditions.     

Electric field allowed molecular transitions for one and two photon excitation microscopy

We propose an excitation technique for observing single and two photon excitation in those molecules for which such transitions are forbidden by the selection rules. This is possible by the application of an external electric field that perturbs the molecular orbitals, thereby resulting in a significant shift of energy levels. Such a shift of energy levels may bring those levels in resonance with the radiation field which is normally forbidden by selection rules. Further, parity of the these states may significantly improve the emission process. The external electric field results in the mixing of excited (short lifetime) and metastable states (long lifetime), thus reducing the lifetime of metastable (or near metastable) states. This may provide an effective channel for allowing transition from the metastable states. An application of electric field may result in the excitation of poorly excitable biomolecules. This excitation technique may find applications in single- and multi-photon fluorescence microscopy, bioimaging and optical devices.     

The Hunt for Ancient Prions: Archaeal Prion-Like Domains Form Amyloid-Based Epigenetic Elements

Prions, proteins that can convert between structurally and functionally distinct states and serve as non-Mendelian mechanisms of inheritance, were initially discovered and only known in eukaryotes, and consequently considered to likely be a relatively late evolutionary acquisition. However, the recent discovery of prions in bacteria and viruses has intimated a potentially more ancient evolutionary origin. Here, we provide evidence that prion-forming domains exist in the domain archaea, the last domain of life left unexplored with regard to prions. We searched for archaeal candidate prion-forming protein sequences computationally, described their taxonomic distribution and phylogeny, and analyzed their associated functional annotations. Using biophysical in vitro assays, cell-based and microscopic approaches, and dye-binding analyses, we tested select candidate prion-forming domains for prionogenic characteristics. Out of the 16 tested, eight formed amyloids, and six acted as protein-based elements of information transfer driving non-Mendelian patterns of inheritance. We also identified short peptides from our archaeal prion candidates that can form amyloid fibrils independently. Lastly, candidates that tested positively in our assays had significantly higher tyrosine and phenylalanine content than candidates that tested negatively, an observation that may help future archaeal prion predictions. Taken together, our discovery of functional prion-forming domains in archaea provides evidence that multiple archaeal proteins are capable of acting as prions—thus expanding our knowledge of this epigenetic phenomenon to the third and final domain of life and bolstering the possibility that they were present at the time of the last universal common ancestor.     

Caecal fermentation characteristics, blood composition and growth of rabbits on substitution of soya-bean meal by unconventional high-glucosinolate mustard ( Brassica juncea) meal as protein supplement

Effect of graded levels of high-glucosinolate mustard (Brassica juncea) meal as substitute of soya-bean meal (SBM) in broiler rabbit diets was studied. Forty weaning rabbits of Soviet Chinchilla and White Giant breed were randomly allocated to one of four experimental diets containing mustard meal (MM) 0, 80, 160 and 245 g/kg. The experiment lasted for 8 weeks. MM had 54.8 mg total glucosinolates (TGLSs) per g dry matter (DM). Diets had TGLS 3.8, 8.4 and 11.98 mg/g DM in 80, 160 and 245 g MM diets, respectively. MM-incorporated diets had higher digestible and linearly (P < 0.01) higher metabolisable energy (ME) content. However, the effect on total tract apparent digestibility of DM, and crude protein was quadratic. Average daily gain (ADG) reduced (P < 0.05) linearly with increasing MM levels in diet, still 80 and 160 g MM diets had similar ADG compared to that of SBM diet. Caecum weight reduced linearly (P < 0.05) with increasing MM levels in diet. The pH of caecal content ranged between 5.85 and 6.19, total N between 1.19 and 1.48 (g per 100 g) and total volatile fatty acids between 4.7 and 5.8 mmol per 100 g, and they were not statistically different. NH3-N ranged between 31.2 and 39.0 mg per 100 ml, and reduced linearly (P < 0.05) while trichloroacetic acid-precipitable nitrogen increased linearly (P < 0.01, ranged between 114 and 247 mg per 100 ml) with increasing MM levels in diet. Blood haemoglobin, packed cell volume and lymphocytes were higher (quadratic effects, P < 0.05) on 245 MM diet, whereas white blood cell count reduced linearly (P < 0.01). Serum aspartate aminotransferase increased linearly (P < 0.01) while alanine aminotransferase and alkaline phosphatase activity, protein, erythrocytes sedimentation rate and red blood cell counts were not affected by MM. Serum Cu, Na and K content increased linearly (P < 0.05) with increasing MM levels. Liver Cu concentration showed quadratic (P < 0.05) increase. Rabbits tolerated 8.4 mg TGLS per g diet (160 g MM per kg) during active growth without any apparent effect on health and growth. It is concluded that MM can replace up to 66% SBM protein in rabbit feeding, whereas complete replacement of SBM with MM reduced feed intake and ADG by 23% and 13%, respectively. Further studies are required to confirm these inclusion levels and glucosinolate tolerance of rabbits.     

Diurnal changes in blood metabolites and their relation to plasma growth hormone and time of feeding in mithun heifers (Bos frontalis)

The aim of the present study was to investigate what, if any, diurnal changes occur in blood metabolites in relation to plasma growth hormone (GH) and feeding time among mithun (Bos frontalis), a semi-wild ruminant. Blood samples were collected at hourly intervals during a 24 h span from 6 mithun heifers (averaging 2.5 yr of age and averaging 230 kg in weight) that were fed twice a day at 11:00 and 16:00 h. Samples were assayed for plasma GH and blood metabolites, non-esterified fatty acids (NEFA), glucose, and alpha-amino nitrogen. The total sampling period was divided into a 1) postprandial (after meal) period (period I: 11:00 to 21:00 h) and 2) interprandial period (period II: 22:00 to 10:00 h) and also into night (20:00 to 05:00 h) and day (06:00 to 10:00 h) periods for statistical analysis. Plasma glucose and alpha-amino nitrogen levels increased (p<0.01), and plasma NEFA and GH decreased (p<0.01) after each meal. No diurnal rhythmicity was detected in plasma glucose or alpha-amino nitrogen levels. Interestingly, plasma NEFA and GH levels were higher (p<0.01) during the interprandial (period II) and night periods, indicating an energy deficit that occurred progressively during the interprandial period of nocturnal feed deprivation. In twice-daily-fed mithuns we conclude that: 1) plasma metabolites and GH exhibited a definite pattern of change with time of feeding; 2) concentrations of plasma NEFA were higher nocturnally due to an energy deficit and that GH levels were higher during the interprandial period after the second meal; 3) the interprandial period after the second feeding may be considered to constitute a short-term food deprivation; 4) the longer interprandial period of 19 h in this study between the second and subsequent morning meal may be changed into equally divided feedings to minimize the short-term energy deficit; and 5) blood sampling for blood metabolites in mithuns should be conducted at a fixed time of day with special emphasis on time of feeding.     

Zidovudine (AZT) treatment suppresses granulocyte-monocyte colony stimulating factor receptor type alpha (GM-CSFR alpha) gene expression in murine bone marrow cells

In vitro exposure of murine bone marrow cells to increasing concentrations of zidovudine (AZT, 0.1-50 microM) had a concentration dependent suppressive effect on the growth of granulocyte-monocyte colony forming unit (CFU-GM) derived colonies. In our previous published study, the mechanism of AZT-induced suppression of erythroid colony forming unit (CFU-E) derived colonies was linked to a decrease in erythropoitin receptor (Epo-R) gene expression. In this study, we have observed that AZT exposure also induced a concentration dependent suppressive effect (35-90%) on GM-CSF receptor type alpha (GM-CSFR alpha) gene expression. The suppression of GM-CSFR alpha mRNA expression was specific, since AZT caused a much lower decrease (15-22%) on the IL-3 receptor type alpha (IL-3R alpha) message level, and had an insignificant effect on glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and c-myc message levels. Erythropoietin (Epo) therapy has been used for reversal of AZT induced erythroid toxicity. Exposure to increasing concentrations (10-500 U/ml) of GM-CSF was unable to override the suppressive effect of AZT on CFU-GM derived colonies, however, treatment in combination with IL-3 (10-250 U/ml) ameliorated the suppressive effects of AZT on CFU-GM and on GM-CSFR alpha and IL-3R alpha gene expression. These findings suggest a mechanism via which AZT may suppress granulocyte-monocyte specific differentiation in murine bone marrow cells. These data also suggest that a combination of GM-CSF and IL-3 may be a superior therapeutic intervention for AZT-induced neutropenia.     

Chronic administration of caffeine: effect on the activities of hepatic antioxidant enzymes of Ehrlich ascites tumor-bearing mice

Chronic ingestion (for 22-30 consecutive days) of caffeine (20 mg/kg/day, p.o.) increased the activities of the hepatic enzymes- catalase (CAT) and superoxide dismutase (SOD) and decreased its lipid peroxidation (LP) in mice. Development of Ehrlich ascites carcinoma (EAC) cell decreased the activities of hepatic CAT and SOD and increased LP. But pretreatment of caffeine for 12 consecutive days and continuation of its treatment during the course of development of EAC cells restored the EAC cell-induced changes in liver CAT, SOD and LP to their corresponding control values. Thus, the present results by confirming the results of others previously published, suggest that caffeine is an antioxidant and may act as an anticarcinogen.