Protective action of salicylic acid against bean yellow mosaic virus infection in Vicia faba leaves

In this study, morphological, ultrastructural and physiological modifications of faba bean (Vicia faba cv Giza 461) leaves in response to bean yellow mosaic virus (BYMV) infection and salicylic acid (SA) treatments were examined. Under BYMV stress, leaves showed symptoms including severe mosaic, mottling, crinkling, size reduction and deformations. Three weeks after virus inoculation, photosynthetic rate, pigment contents and transpiration rate were significantly reduced in response to BYMV infection.

Ultrastructural investigations of BYMV-infected leaves demonstrated that most chloroplasts with increased stromal area became spherical in shape and some lost their envelopes, either partially or totally. The internal structures of chloroplast, grana and thylakoids were dilated. Two kinds of inclusions were detected in BYMV-infected leaves: straight or slightly curved bands sometimes coiled or looped at the end, and electron opaque crystals with varied shapes. BYMV-infected cells showed lower chloroplast number in comparison to the control.

Spraying of SA on faba bean leaves helped to reduce or prevent the harmful effects produced after virus infection. Application of 100 μM SA three days before inoculation restored the metabolism of infected leaves to the levels of healthy controls. SA treatment improved plant health by increasing the photosynthesis rates, pigment contents and levels of other parameters studied similar to control values.

Moreover, SA treatment increased plant resistance against BYMV. This was observed through induction of chloroplast number, reduction in percentage of infected plants, decrease in disease severity and virus concentration of plants treated with SA prior to BYMV inoculation. Cells of SA-treated samples showed well-developed chloroplasts with many starch grains and well-organized cell organelles.

The present results provide an overview of the negative effects on faba bean leaves due to BYMV infection from physiological and subcellular perspectives. Also, a role of SA involved in induction of resistance against BYMV infection in bean plants is discussed.     

Three new species of Neoechinorhynchus (Acanthocephala: Neoechinorhynchidae) from fresh-water fish collected from Tigris River,Iraq

Systematic Parasitology - Three new species of Neoechinorhynchus are described from freshwater fish of Tigris River, at Salah Al-Din province, mid Iraq, namely Neoechinorhynchus tigrisensis n. sp....     

Synthesis of biologically active steroid derivatives by the utility of Lawesson's reagent

Steroid derivatives V, VI, VII and VIII reacted with Lawesson's reagent (LR) to produce spiro-oxazaphosphole-4',17-androstene derivative XI, diazaphospholoandrostane XIV and the thionated derivatives XVI and XVII, respectively. The structures of the new compounds were confirmed by analytical and spectroscopic evidence. A mechanism accounting for the formation of the new compounds was given. The in vitro antimicrobial activity of the new compounds were tested.     

DISCRETE MEMBRANOUS SUBAORTIC STENOSIS: POSTOPERATIVE REVERSION OF AORTIC VALVE ECHOCARDIOGRAM TO NORMAL

Clinical, hemodynamic, angiographic and echocardiographic findings in a patient with discrete membranous subaortic stenosis are presented. Echocardiography revealed evidence of a normal mitral valve and abnormal movement of the aortic valve with poor motion and fluttering of the anterior cusp. After a subvalvular membrane subjacent to the aortic valve cusp was excised, postoperative echocardiographic study showed normal movement of the aortic valve.     

Exogenously applied ascorbic acid ameliorates detrimental effects of NaCl and mannitol stress in Vicia faba seedlings

The adverse effects of either NaCl or mannitol on growth, nitrogen content, and antioxidant system in Vicia faba seedlings were investigated. The role of exogenous ascorbic acid in increasing resistance to these stressors was also evaluated. Thus, with an increase in concentration of either NaCl or mannitol in culture media, a progressively greater significant decrease in percentage germination, in growth parameters, and in nitrogen constituents of the germinating beans, was observed. On the other hand, amide-, nitrate-, and total soluble-N contents appeared to show a progressive significant increase. Exogenous addition of ascorbic acid (4 mM) to the stressful media induced a pronounced significantly increased percentage germination and the growth attributes, whereas nitrogen constituents were variably changed in relation to values maintained in beans treated with either NaCl or mannitol. Furthermore, exogenous addition of ascorbic acid to NaCl or mannitol media induced a significant increase in the contents of ascorbate and glutathione and enzymatic antioxidant activities, in particular, in beans treated with the three highest concentrations of NaCl or mannitol, throughout the period of the experiments (12 days). Thus, ascorbic acid ameliorates the adverse effects of the stressful media; the magnitude of amelioration being a function of the type and the concentration of the stressful agent as well as of the duration of treatment. The importance of the above-mentioned changes in growth and metabolism to stress tolerance in broad bean is discussed.     

Histopathological and biochemical changes of morphine sulphate administration on the cerebellum of albino rats

In this study the long-term effects of morphine sulphate treatment (MST) on histopathological and biochemical changes in the cerebellum was assessed in albino rats. Normal saline (5 ml) was given orally as placebo in the control group (n = 25). Morphine groups received morphine orally at a dose level of 5 mg/kg body weight day after day for 10, 20 and 30 days (n = 25/group). Light microscopy revealed that the molecular layer showed vacuolation. The Purkinje cells lost their specific shaped appearance, decreased in size and numbers. The granular cells highly degenerated. Electron microscopy revealed fragmentation of the cisterns of the both types of endoplasmic reticulum, resulted in a progressive depletion of total protein contents as well as general carbohydrates in all treated groups as supported by histochemical observation. Obvious destruction of mitochondrial inner membrane and cristae mediate cell death. Also, abnormal nucleus with deformed perforated nuclear membrane and deformation of the plasma membrane with degeneration of the synapses could interpreted as a sign of necrosis. Biochemical analysis revealed that dopamine (DA) and norepinephrine (NE) were significantly decreased in four brain areas (cortex striatum, thalamus/hypothalamus, and cerebellum). In contrast, serotonin (5-HT) level was increased in these brain regions; with an exception of 5-HT on day 10 and neurotransmitter levels in the pons were unaffected. The quantitative analysis showed a significant decrease (P < 0.05) in the diameter of Purkinje cells and in the thickness of both molecular and granular layers treated groups. Morphine sulphate induces may be a cell death or necrosis in the rat cerebellum and modulating neurotransmitter system. Our findings pointed out the risk of increased cerebellum damage due to long-term of morphine use.     

Modifications of antioxidant activity and protein composition of bean leaf due to <Emphasis Type=Italic>Bean yellow mosaic virus</Emphasis> infection and salicylic acid treatments

The antioxidant status as well as protein composition of faba bean leaves infected with Bean yellow mosaic virus (BYMV) and the effect of salicylic acid application was examined in this paper. Some modifications in the antioxidant status were observed by changing some antioxidant enzymes activities and contents of antioxidant metabolites. BYMV-infected leaves revealed POD, CAT, APX and SOD induced activities while SA treatments could inhibit POD, CAT activities but induced SOD activity. The enzyme activities seemed to be SA concentration dependant. Higher H₂O₂ and MDA concentrations were recorded in virus-infected leaves than those of the corresponding controls while treatment with SA followed by virus inoculation caused lowering of MDA concentration and reducing the damage due to lipid peroxidation. Moreover, because of virus infection and/or SA treatments, an increase in the amounts of phenolics and flavonoids was noticed. As compared to the control, BYMV infection or SA application caused pronounced increase in the antioxidant activity of leaf extracts detected by DPPH assay, indicating an increase in the amounts of antioxidant compounds occurred. To test the protein composition, the contents of each protein fractions (soluble, insoluble and total) were analyzed and the change in protein patterns was visualized using SDS-PAGE. The BYMV-infected bean leaves had protein contents higher than the control indicating accumulation of pathogenesis-related proteins. Moreover, spraying SA with or without virus inoculation could accumulate soluble, insoluble and total proteins and the pattern of increase was in accordance with SA concentration. Alterations in protein patterns were observed in faba bean leaves (Vicia faba cv Giza 461) in response to BYMV infection and SA treatments. Because of BYMV infection and SA treatments, the protein profiles showed new bands in comparison to the control. Some polypeptides were highly accumulated in treatments of SA followed by virus inoculation. Changing antioxidant status and accumulation of some antioxidant metabolites as well as the pronounced alterations in the protein composition indicate a kind of plant response against pathogen invasion and in case of SA treatment it is considered a way by which a defence response was initiated and/or activated.     

β-Glucosidase catalyzed synthesis of octyl-β-D-glucopyranoside using whole cells of Pichia etchellsii in micro aqueous media

Octyl-β-D-glucopyranoside was synthesized by transglucosylation between p-nitrophenyl β-D-glucopyranoside (pNPG) and octanol as an acceptor using whole cells of thermo tolerant yeast Pichia etchellsii displaying cell wall bound β-glucosidase. Effect of several parameters such as glucosyl donor concentration, enzyme units and initial water activity was studied to optimize product yield. An initial water activity interval of 0.33-0.64 was favorable and increase in total enzyme units had marginal effect on conversion yield. An empirical model was developed to describe the relationship between various parameters and octyl glucoside yield. These factors were combined in a batch replacement strategy whereby octyl-β-D-glucopyranoside was synthesized in 4h to a concentration of 30 mM (9.25 mg/ml) with a conversion yield of nearly 70% with pNPG as a glucosyl donor. Quantitative analysis was done by a highly reproducible reverse-phase high-performance liquid chromatography (RP-HPLC) method and detection was achieved using refractive index detector. The structure of the product was confirmed by 13C and 1H NMR spectroscopy. Additional products like octyl diglucoside were also formed, the structure of which was confirmed by mass spectrometry.     

Staphylococcus aureus strains isolated from bovine mastitis: virulence,antibody production and protection from challenge in a mouse model

Septic arthritis in mice was used as a model to evaluate the virulence of Staphylococcus aureus and coagulase-negative staphylococci (CNS) isolated from cases of bovine mastitis. In addition, the model was used to evaluate the cross protection elicited by heterologous antibodies. Mice were intramuscularly inoculated with serial bacterial doses of different strains of S. aureus or CNS, for virulence determination; they were monitored for arthritis, gangrene or death up to 20 days. Antibody response, cross reactivity and resistance to challenge were tested by subcutaneous inoculation with a low dose of one of the S. aureus or CNS strains followed by challenge with two S. aureus strains. S. aureus alpha-hemolysin isolate was the most virulent, followed by alpha+beta-hemolysin and beta-hemolysin isolates. The least virulent isolates were the non-hemolytic S. aureus strains but even they were more virulent than the CNS strains tested. Antibodies against three different S. aureus antigens were detected by the ELISA in all mice that were inoculated with the S. aureus strains but not in any of those with the CNS strains. Immunoblot test against various S. aureus strains as antigens showed high cross-reactivity among the S. aureus strains but only a slight similarity, restricted to the bands above 36 kDa, with the CNS sera. Low-dose inoculation of alpha or alpha+beta strains before challenge with homologous and heterologous strains protected the mice, whereas the two beta strains provided only partial protection. The inoculations of non-hemolytic S. aureus or the CNS strains did not elicit any protection. Our findings demonstrate that pre-exposure of mice to a low dose of certain S. aureus strains could provide protection and that the antibodies produced could have an important protective role.     

The insecticide DDT targets the OSCP and subunit D of the Apis mellifera ATP synthase

1,1-bis (p-chlorophenyl)-2, 2, 2-trichloroethane (DDT) has been used for control of malaria mosquitoes and other insect vectors of human diseases since 1945. Its use poses an environmental dilemma and efforts to replace it have been hampered by lack of information about its molecular target. This work identifies the 23 kDa band responsible for the DDT sensitivity in bees, as the OSCP and subunit "d" of the ATP synthase. The OSCP of the bee's ATP synthase contained 207 amino acids compared to 190 in bovine, which is insensitive to DDT, and the identities were only 47%. Subunit "d" of the bees had no counterpart in the bovine. Whether DDT is interacting only with OSCP, only with subunit "d", or with both subunits, remains to be assessed. Identification of the molecular target of DDT will lead the way to new target based insecticides aimed to protect plant, combat malaria and other insect transmitted diseases.