Effects of Parenteral Supply of Iron on RBC Parameters,Performance, and Health in Neonatal Dairy Calves

The present study was designed to evaluate the effects of parenteral iron on red blood cell parameters, performance, and health in dairy Holstein calves. Twenty neonatal calves were equally divided at random into two groups, one of which served as controls. Care was taken to ensure homogeneity of sex, age, and general health status of the animals. The controls received a normal diet and water ad libitum, while the study animals were injected with 1 g iron as Fe-dextran 2 days after birth. A daily record was kept of the calves’ weight and growth parameters. At periods of 24–48 h after birth and at 14, 28, and 42 days of age, jugular blood was drawn from all the experimental and control animals to measure the packed cell volume, red blood count, hemoglobin, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and iron levels. At the start of the experiments, there were no significant differences between these parameters between the two study groups (p > 0.05). With time, significant differences were seen between most of the values measured (p < 0.05) except for the mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and iron level. Significant differences were seen for total weight gain and mean daily weight gain, which were higher in the iron-supplemented group (p < 0.05).     

Polyphenol oxidase activity in the roots of seedlings of Bromus (Poaceae) and other grass genera

? Premise of the study: Phenolic compounds exuded by roots have been implicated in allelopathic interactions among plants. Root enzymes that destroy phenolics may protect plants against allelopathic inhibition and thus may aid in invasiveness. Phenolic-degrading enzymes are chiefly found in aboveground plant parts, but have also been previously reported in root tissues where the enzyme's function is unknown. We explored phenolic oxidase activity in emerging roots of grasses in a survey across different grass genera; in particular, we aimed to test whether grasses of the genus Bromus, known for their large invasion potential, differ in this respect from other grass taxa. ? Methods: We assayed a range of grass genera commonly found in the United States for root enzyme activity with spectrophotometric assays of phenol oxidase activity using l-DOPA as the main substrate. ? Key results: In the survey of a grass genera, we discovered that roots of the genus Bromus contain large amounts of polyphenol oxidase (PPO) activity, while all other tested grass genera, even ones closely related to Bromus, did not. PPO was found to be present at germination and remained active throughout the life of the plant. Compared to other PPOs, the enzyme present in Bromus appears to have a narrow substrate range. ? Conclusions: The specific functions of the root PPO and the ecological ramifications of the special status of Bromus are not yet clear. The possibility that the enzyme plays a role in plant species interaction for bromes, a genus of grasses known to have high invasive potential, is raised.     

Investigation on Mitochondrial tRNA<Superscript>Leu/Lys</Superscript>, NDI and ATPase 6/8 in Iranian Multiple Sclerosis Patients

As with chromosomal DNA, the mitochondrial DNA (mtDNA) can contain mutations that are highly pathogenic .In fact, many diseases of the central nervous system are known to be caused by mutations in mtDNA. Dysfunction of the mitochondrial Respiratory Chain (RC) has been shown in patients with neurological disease including Alzheimer’s disease (AD), Parkinson’s disease (PD) and Multiple sclerosis (MS). MS is a demyelinating disease of central nervous system characterized by morphological hallmarks of inflammation, demyelination and axonal loss. Considering this importance, we decided to investigate several highly mutative parts of mtDNA for point mutations as MT-LTI (tRNA^{Leucine1(UUA/G)}), MT-NDI (NADH Dehydrogenase subunit 1), MT-COII (Cytochrome c oxidase subunit II), MT-TK (tRNA^Lysine), MT-ATP8 (ATP synthase subunit F0 8) and MT-ATP6 (ATP synthase subunit F0 6) in 20 Iranian MS patients and 80 age-matched control subjects by PCR and automated DNA sequencing to evaluate any probable point mutations. Our results revealed that 15 (75%) out of 20 MS patients had point mutations. Some of point mutations were newly found in this study. This study suggested that point mutation occurred in mtDNA might be involved in pathogenesis of MS.     

A theoretical elucidation of bilirubin interaction with HSA's lysines: first electrostatic binding site in IIA subdomain

The electrostatic interaction of amino acid lysines 190, 195 and 199 of human serum albumin (HSA) with bilirubin have been investigated using molecular dynamic simulations, QM and QM/MM minimization methods. In this study two methodological approaches have been employed. In the first approach X-ray structure and the structure obtained from the molecular dynamic simulation of subdomain IIA of HSA in vacuum have been utilized. Interactions have been evaluated with the segment 186-200 of the cited subdomain. Calculations on the X-ray structure of above segment indicate an effective interaction of the lysine 195 with bilirubin, although that of the lysine 190 is also found considerable in this structure. Performing simulation in vacuum, it has been revealed that except for the lysine 195, the other two lysine residues (190 and 199) could not be considered as centers of interaction. Such finding, which is in accord with experimental data, lends support to the procedure employed in this study. NBO analyses suggest that tasks to achieve a structure indicating bilirubin interaction with the lysine 195 from the 186-200 segment extracted from X-ray structure, results in a structure that lacks any electrostatic interaction. In fact, it has been found that the stability of the latter species can be attributed to the H-bonding interaction of the glutamate 188 with both bilirubin and the lysine 195. Further NBO analysis on the structure of the same species, while achieved after molecular dynamic simulation on subdomain IIA in vacuum has revealed that a favorable electrostatic interaction between the lysine 195 and bilirubin has occurred. Besides, H-bonding interaction of the glutamate 188 with bilirubin has been evident in the same species. For the second approach, presence of water molecules and ions has been considered to simulate condensed medium. Applying docking, conformational sampling, and QM/MM minimization steps in sequence, a structure has been achieved which presents a specific interaction between epsilon-NH3(+) group of the lysine 195 residue and the lactam oxygen atom of bilirubin. NBO analyses suggest that above electrostatic interaction is combined with hydrogen bonding interaction between same two groups. Moreover, a hydrogen bond between oxygen atom of bilirubin's acetate group and alpha-NH group of lysine 195 has been observed. Molecular orbital calculations have been presented which support the NBO analyses.     

Chemotaxonomic significance of the essential oils of 18 Ferula species (Apiaceae) from Iran

To evaluate the chemotaxonomic significance of the essential oils of 23 populations of 18 Iranian Ferula species, the chemical composition of the oils was investigated by GC/FID and GC/MS. Altogether, 84 constituents, representing 81.3-99.7% of the total composition of the oils, have been identified. The composition of six species of the genus, i.e., F. oopoda, F. foetida, F. behboudiana, F. diversivittata, F. galbaniflua, and F. hezarlalehzarica, has been reported for the first time. The main constituents identified were α-terpinyl acetate (73.3%), 2,3,4-trimethylthiophene (2; 49.0%), sabinene (75.3%), verbenone (5; 69.4%), β-pinene (59.0-66.3%), and (Z)-β-ocimene (41.7%). Cluster analysis (CA) of the percentage content of the essential oil components of the Ferula species resulted in the characterization of four groups, i.e., taxa containing either i) monoterpene hydrocarbons, ii) oxygenated monoterpenes, iii) organosulfur compounds, or iv) monoterpene, sesquiterpene, and aliphatic hydrocarbons as the principal classes of compounds. Based on the results obtained, the chemical independence of F. hirtella from F. szowitsiana and of F. galbaniflua from F. gummosa at the specific level was concluded and their positions as distinct species were confirmed. The chemotaxonomic relationships among the representatives of the genus Ferula have been discussed in detail.     

Angiotensin II increases the permeability and PV-1 expression of endothelial cells

Angiotensin II (ANG II), the major effector molecule of the renin-angiotensin system (RAS), is a powerful vasoactive mediator associated with hypertension and renal failure. In this study the permeability changes and its morphological attributes in endothelial cells of human umbilical vein (HUVECs) were studied considering the potential regulatory role of ANG II. The effects of ANG II were compared with those of vascular endothelial growth factor (VEGF). Permeability was determined by 40 kDa FITC-Dextran and electrical impedance measurements. Plasmalemmal vesicle-1 (PV-1) mRNA levels were measured by PCR. Endothelial cell surface was studied by atomic force microscopy (AFM), and caveolae were visualized by transmission electron microscopy (TEM) in HUVEC monolayers. ANG II (10(-7) M), similarly to VEGF (100 ng/ml), increased the endothelial permeability parallel with an increase in the number of cell surface openings and caveolae. AT1 and VEGF-R2 receptor blockers (candesartan and ZM-323881, respectively) blunted these effects. ANG II and VEGF increased the expression of PV-1, which could be blocked by candesartan or ZM-323881 pretreatments and by the p38 mitogem-activated protein (MAP) kinase inhibitor SB-203580. Additionally, SB-203580 blocked the increase in endothelial permeability and the number of surface openings and caveolae. In conclusion, we have demonstrated that ANG II plays a role in regulation of permeability and formation of cell surface openings through AT1 receptor and PV-1 protein synthesis in a p38 MAP kinase-dependent manner in endothelial cells. The surface openings that increase in parallel with permeability may represent transcellular channels, caveolae, or both. These morphological and permeability changes may be involved in (patho-) physiological effects of ANG II.     

Investigation of tRNA<Superscript>Leu/Lys</Superscript> and ATPase 6 Genes Mutations in Huntington’s Disease

Huntington disease (HD) is a genetically dominant condition caused by expanded CAG repeats which code for glutamine in the HD gene product, huntingtin. Huntingtin is expressed in almost all tissues, so abnormalities outside the brain can also be expected. Involvement of nuclei and mitochondria in HD pathophysiology has been suggested. In fact mitochondrial dysfunction is reported in brains of patients suffering from HD. The tRNA gene mutations are one of hot spots that can cause mitochondrial disorders. In this study, possible mitochondrial DNA (mtDNA) damage was evaluated by screening for mutations in the tRNA^leu/lys and ATPase 6 genes of 20 patients with HD, using PCR and automated DNA sequencing. Mutations including an A8656G mutation in one patient were observed, which may be causal to the disease. Understanding the role of mitochondria in the pathogenesis of neurodegenerative diseases could potentially be important for the development of therapeutic strategies in HD.     

Adapting pharmacokinetic properties of a humanized anti-interleukin-8 antibody for therapeutic applications using site-specific pegylation.

A neutralizing anti-interleukin-(IL-)8 monoclonal antibody was humanized by grafting the complementary determining regions onto the human IgG framework. Subsequent alanine scanning mutagenesis and phage display enabled the production of an affinity matured antibody with a >100-fold improvement in IL-8 binding. Antibody fragments can be efficiently produced in Escherichia coli but have the limitation of rapid clearance rates in vivo. The Fab' fragment of the antibody was therefore modified with polyethylene glycol (PEG) in order to obtain a more desirable pharmacokinetic profile. PEG (5-40 kDa) was site-specifically conjugated to the Fab' via the single free cysteine residue in the hinge region. In vitro binding and bioassays showed little or no loss of activity. The pharmacokinetic profiles of the 20 kDa, 30 kDa, 40 kDa, and 40 kDa branched PEG-Fab' molecules were evaluated in rabbits. Relative to the native Fab', the clearance rates of the PEGylated molecules were decreased by 44-175-fold. In a rabbit ear model of ischemia/reperfusion injury, all PEGylated Fab' molecules were as efficacious in reducing oedema as the original monoclonal antibody. These studies demonstrate that it is possible to customize the pharmacokinetic properties of a Fab' while retaining its antigen binding activity.     

Tetraploid induction in savory (Satureja khuzistanica): cytological,morphological, phytochemical and physiological changes

Plant Cell, Tissue and Organ Culture (PCTOC) - Satureja khuzitanica, due to having phenolic compounds such as carvacrol in its essential oil and rosmarinic acid in its extract has strong...