Branched chain amino acids induce apoptosis in neural cells without mitochondrial membrane depolarization or cytochrome c release: implications for neurological impairment associated with maple syrup urine disease

Maple syrup urine disease (MSUD) is an inborn error of metabolism caused by a deficiency in branched chain alpha-keto acid dehydrogenase that can result in neurodegenerative sequelae in human infants. In the present study, increased concentrations of MSUD metabolites, in particular alpha-keto isocaproic acid, specifically induced apoptosis in glial and neuronal cells in culture. Apoptosis was associated with a reduction in cell respiration but without impairment of respiratory chain function, without early changes in mitochondrial membrane potential and without cytochrome c release into the cytosol. Significantly, alpha-keto isocaproic acid also triggered neuronal apoptosis in vivo after intracerebral injection into the developing rat brain. These findings suggest that MSUD neurodegeneration may result, at least in part, from an accumulation of branched chain amino acids and their alpha-keto acid derivatives that trigger apoptosis through a cytochrome c-independent pathway.     

A preliminary study of human paraoxonase and PON 1 L/M55–PON 1 Q/R 192 polymorphisms in Turkish patients with coronary artery disease

Paraoxonase 1 (PON 1) is a high‐density lipoprotein (HDL)‐associated enzyme with antioxidant function protecting low‐density lipoprotein (LDL) from oxidation. PON 1 has two amino acid polymorphisms in coding region; L/M 55 and Q/R 192. These polymorphisms modulate paraoxonase activity of the enzyme. PON 1 activity decreases in coronary artery disease (CAD). In the present study, distribution of PON 1 L/M 55 and Q/R 192 polymorphisms and the effect of these polymorphisms on the activities of PON 1, and on the severity of CAD in 277 CAD (+) patient and 92 CAD (?) subjects were examined. PON 1 L/M 55 and Q/R 192 genotypes were determined by PCR, RFLP and agarose gel electrophoresis techniques. Genotype distributions and allele frequencies for PON 1 Q/R 192 polymorphism were not significantly different between controls and CAD (+) patient group (p > 0.05), but in genotype and allele distribution of PON 1 L/M55 polymorphism, there was significantly difference among groups (p < 0.05). Genotype distributions for both polymorphisms were not significantly different between subgroups of single‐vessel disease (SVD), double‐vessel disease (DVD) and triple‐vessel disease (TVD). Serum PON 1 activity was lower in CAD (+) group than in controls and this was also statistically significant (p < 0.001). In both groups, the highest PON activities were detected in LL and RR genotypes. In summary, our results suggest that there is an association between the PON 1 L/M 55 polymorphism of paraoxonase and CAD in Turkish patients but not with PON 1 Q/R 192 polymorphism. However, it is hard to correlate these polymorphisms and severity of CAD. Copyright © 2009 John Wiley & Sons, Ltd.     

Polyaniline grafted polyacylonitrile conductive composite fibers for reversible immobilization of enzymes: Stability and catalytic properties of invertase

Polyacylonitrile fibers (PAN) surfaces were modified with chemical polymerization of conductive polyaniline (PANI) in the presence of potassium dichromate as an oxidizing agent. The effect of aniline concentration on the grafting efficiency and on the electrical surface resistance of PAN/PANI composite fibers was investigated. The surface resistance of the conductive composite fibers in this work was found to be between 8.0 and 0.5 kΩ/cm. As the amount of grafted PANI increased on the PAN fibers the electrical resistance of composite fibers decreased. The PAN/PANI composite fibers were characterized by SEM and FTIR studies. Composite PAN/PANI fibers were used for reversible immobilization of invertase. The immobilization efficiency and the activity of the immobilized invertase (from 1.0 mg/mL invertase solution at pH 5.5) were increased with increasing PANI contents of the composite fibers. The maximum amount of immobilized enzyme onto composite fibers containing 2.0% PANI was about 76.6 mg/g. The optimum pH for the free enzyme was observed at 5.0. On the other hand, immobilized invertase yielded a broad optimum pH profile between pH 5.0 and 7.0. Immobilized invertase exhibited 83% of its original activity even after two months storage at 4 °C while the free enzyme showed only 7% of its initial activity.     

Effect of EDTA and Tannic Acid on the Removal of Cd,Ni, Pb and Cu from Artificially Contaminated Soil by Althaea rosea Cavan

In this study an ornamental plant of Althaea rosea Cavan was investigated for its potential use in the removal of Cd, Ni, Pb and Cu from an artificially contaminated soil. Effect of two different chelating agents on the removal has also been studied by using EDTA (ethylenediaminetetracetic acid) and TA (tannic acid). Both EDTA and TA have led to higher heavy metal concentration in shoots and leaves compared to control plants. However EDTA is generally known as an effective agent in metal solubilisation of soil, in this study, TA was found more effective to induce metal accumulation in Althaea rosea Cavan under the studied conditions. In addition to this, EDTA is toxic to some species and restraining the growth of the plants. The higher BCF (Bio Concentration Factor) and TF (Translocation Factor) values obtained from stems and leaves by the effects of the chemical enhancers (EDTA and TA) show that Althaea rosea Cavan is a hyper accumulator for the studied metals and may be cultivated to clean the contaminated soils.     

Fatal Systemic Cladosporiosis in a Merino Sheep Flock

Systemic cladosporiosis is described in 25 merino sheep from a flock consisting 250 animals. The fungal pneumonia appeared after an intensive antibiotic treatment, because of a respiratory system disorder. The pen of the flock was humid and crowded, and animals had signs of respiratory distress, coughing, fever and anorexia. All of the ill animals died, and necropsy was performed on 10 sheep. The lesions were characterized by a multifocal pyogranulomatous pneumonia and an abomasitis. Severe hemorrhages were observed in the lungs. At the histopathological examination, severe vasculitis with thrombosis was observed in various organs, especially in the lungs and abomasums, suggestive for a hematogenous dissemination of the infection in these organs. Numerous PAS-positive fungal elements were seen in the pyogranulomatous foci. Dark green fungal colonies were seen in the blood agar and Sabouraud dextrose agar that were identified as Cladosporium cladosporioides. This report highlights that phaeohyphomycosis can cause a severe systemic and fatal disease in merino sheep under insufficient management conditions.     

Antimicrobial susceptibilities of Coagulase-Negative Staphylococci (CNS) and Streptococci from bovine subclinical mastitis cases

The prevalence and antimicrobial susceptibilities of Staphylococci and Streptococci were assessed from subclinical mastitis cases. One hundred Coagulase-Negative Staphylococci (CNS) and 34 Streptoccocci were identified. The most frequently isolated species were Staphylococcus haemolyticus (27%) and Staphylococcus simulans (24%). Susceptible CNS species revealed the highest resistance to penicillin G (58%), ampicillin (48%), neomycin (20%), and oleandomycin (14%). CNS methicillin resistance rates within 82 isolates were 21.95% and 1.22% by disk diffusion and PCR methods, respectively. These results suggested the disk diffusion method was more prone to yield false positives. Partial sequencing of the 16S rRNA region from the mecA carrying isolate (S. haemolyticus) was homologous with S. haemolyticus sequences/accessions obtained from GenBank. However, the mecA gene sequence from this isolate was more closely allied with the S. aureus mecA gene of human origins. Identical sequence data was acquired from the National Center for Biotechnology Information (NCBI) database, suggesting horizontal gene transfer between the two species. CNS β-lactamase activity within 81 isolates was 29.63%. The most frequently isolated Streptococcus species were S. uberis (52%) and S. agalactiae (15%). Oleandomycin was the least effective antimicrobial agent on these isolates with 59% susceptibility. Results indicated that CNS and Streptococci exhibited various antimicrobial resistance responses. Consequently, isolation and identification of udder pathogens in herds suffering from subclinical agents is essential to select the most effective antimicrobial agent. Moreover, multiple resistance features of methicillin resistant (MR) isolates should be considered during antimicrobial susceptibility tests.     

Antifungal activity of essential oils against three vegetative-compatibility groups of <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

The antifungal activities of volatile phase effects of essential oils from Origanum onites, O. syriacum, O. minutiflorum, O. vulgare, O, marjorana, Thymus vulgaris, T. serpyllum, Rosmarinus officinalis, Salvia officinalis and Micromeria fruticosa were evaluated for their ability to inhibit growth of three vegetative compatibility groups (VCGs) of Verticillium dahliae. Carvacrol was the main component of O. onites, O. minutiflorum and O. vulgare essential oils, while γ-terpinene was the main component of O. syriacum. P-cymene and thymol were the dominant component of T. vulgaris and T. serpyllum. β- thujone and l-camphor were the main component of S. officinalis. Polegone and isomenthone were the dominant components of M. fruticosa essential oil. Based on the in vitro test, the degree of fungistatical effects can be ranked in the following order of inhibition: O. syriacum = O. onites = O. minutiflorum = O. vulgare = T. vulgaris > T. serpyllum > M. fruticosa > S. officinalis = O. marjorana > R. officinalis. The essential oils of S. officinalis, O. marjorana and R. officinalis displayed moderate antifungal activity, that increased with increasing concentrations. Among the VCGs, VCG2A and VCG4B were found to be highly sensitive to the essential oils. The essential oils of O. syriacum, O. onites, O. minutiflorum, O. vulgare and T. vulgaris were the most efficacious, demonstrating strong antifungal activity against all of the tested VCGs of V. dahliae at relatively low concentrations and they could find practical application as natural fungicides in the prevention and protection of plants from V. dahliae infections.     

Protective effect of quercetin on liver damage induced by biliary obstruction in rats

The aim of this study was to evaluate the possible protective effects of quercetin (QE) against cholestatic oxidative stress and liver damage in the common bile duct ligated rats. A total of 24 male Wistar albino rats were divided into three groups: control, bile duct ligation (BDL) and BDL + received QE; each group contain 8 animals. The rats in QE treated groups were given QE (15 mg/kg) once a day intraperitoneally for 4 weeks starting 3 days prior to BDL operation. The changes demonstrating the bile duct proliferation and fibrosis in expanded portal tracts include the extension of proliferated bile ducts into lobules, mononuclear cells, and neutrophil infiltration into the widened portal areas were observed in BDL group. Treatment of BDL with QE attenuated alterations in liver histology. The alpha smooth muscle actin (α-SMA), transforming growth factor beta (TGF-β1) positive cells and the activity of TUNEL in the BDL were observed to be reduced with the QE treatment. The data indicate that QE attenuates BDL-induced cholestatic liver injury, bile duct proliferation, and fibrosis. The hepatoprotective effect of QE is associated with antioxidative potential.     

Morphology, anatomy, palynology and nutlet micromorphology of the rediscovered Turkish endemic Salvia ballsiana (Lamiaceae) and their taxonomic implications

Salvia ballsiana (Rech. fil.) Hedge, previously known only from the type gathering, is a local endemic species from Turkey. The species was first collected from Gölbaşı (Malatya) by E. K. Balls in 1935, and was not collected again until 2008, when we found it in Gerger (Adıyaman). The diagnostic morphological characters of this rare endemic species are discussed, including its anatomical, palynological and nutlet micromorphological features. Morphological characteristics of leaves, calyces, corollas and types of stamens are useful for sectional and specific delimitation in Salvia . Anatomical characters such as number of ray rows in roots, distance between vascular bundles in stems, mesophyll structures in leaves, shape of mid-rib and presence/absence of sclerenchymatic tissue in petioles are of taxonomic significance. In addition, size, shape and ornamentation of pollen grains and nutlets are diagnostic.