Recovery of antioxidant phenolics from white vinification solid by-products employing water/ethanol mixtures

Solid wastes from white vinification, including grape peels, seeds and stems, were used as raw material for the recovery of antioxidant polyphenols. Extractions were performed using non-toxic media composed of water/ethanol mixtures and hydrochloric, acetic or tartaric acid. Recovery efficiency was assessed by monitoring the antioxidant potency of extracts and several indices related to their polyphenolic composition, including total polyphenol, total flavonoid, total flavanol and condensed tannin (proanthocyanidin) content. Among the by-products tested, seeds were shown to contain exceptional amounts of total polyphenols (13.76 g per 100g dry weight), followed by stems (7.47 g per 100g dry weight) and peels (0.97 g per 100g dry weight). Extracts with the highest antioxidant activity from all by-products were obtained with 57% ethanol. Acidification of this medium with 0.1% HCl improved polyphenol recovery and antiradical activity for stem extracts, but it was unfavourable for seed extraction.     

Terrestrial slugs (Gastropoda, Pulmonata) in the NATURA 2000 areas of Cyprus island

Terrestrial slugs of the Island of Cyprus were recently studied in the framework of a study of the whole terrestrial malacofauna of the island. The present work was carried out in the Natura 2000 conservation areas of the island in 155 sampling sites over three years (2004-2007). Museum collections as well as literature references were included. In total six species are present in the Natura 2000 areas of the island, belonging to three families: Limacidae, Agriolimacidae and Milacidae. One of the species, Milax riedeli, is a new record for the island. The distribution of the species across the island and in the surrounding areas is discussed.     

Inosine induced mutations

Summary Two complementary 24 base single stranded oligonucleotides containing randomly located inosine residues were synthesized in vitro. Once annealed, the two oligonucleotides were cloned into derivatives of ColE1 and transformed into Escherichia coli. Sequence analysis of 157 clones yielded 305 mutations. The pattern of the mutations revealed the following: (1) The frequency of inosine induced mutations was significantly less than predicted from its content in the oligonucleotides; (2) Inosine incorporation resulted almost exclusively in base changes to guanine; (3) The mutation distribution is biased towards A/T to G/C substitutions; (4) There were reproducible position biases; and (5) There was a reproducible strand bias which was independent of the cassette orientation with respect to the plasmid origin of replication.     

COPD exacerbation: Lost in translation

The introduction and acceptance of a standard definition for exacerbations of COPD can be helpful in prompt diagnosis and management of these events. The latest GOLD executive committee recognised this necessity and it has now included a definition of exacerbation in the guidelines for COPD which is an important step forward in the management of the disease. This definition is pragmatic and compromises the different approaches for exacerbation. However, the inclusion of the "healthcare utilisation" approach (".. may warrant a change in regular medication") in the definition may introduce in the diagnosis of exacerbation factors related to the access to health care services which may not be related to the underlying pathophysiologal process which characterizes exacerbations. It should be also noted that the aetiology of COPD exacerbations has not yet been included in the current definition. In this respect, the definition does not acknowledge the fact that many patients with COPD may suffer from additional conditions (i.e. congestive cardiac failure or pulmonary embolism) that can masquerade as exacerbations but they should not be considered as causes of them. The authors therefore suggest that an inclusion of the etiologic factors of COPD exacerbations in the definition. Moreover, COPD exacerbations are characterized by increased airway and systemic inflammation and significant deterioration in lung fuction. These fundamental aspects should be accounted in diagnosis/definition of exacerbations. This could be done by the introduction of a "laboratory" marker in the diagnosis of these acute events. The authors acknowledge that the use of a test or a biomarker in the diagnosis of exacerbations meets certain difficulties related to performing lung function tests or to sampling during exacerbations. However, the introduction of a test that reflects airway or systemic inflammation in the diagnosis of exacerbations might be another step forward in the management of COPD.     

Alteration of P450 distal pocket solvent leads to impaired proton delivery and changes in heme geometry

Distal pocket water molecules have been widely implicated in the delivery of protons required in O-O bond heterolysis in the P450 reaction cycle. Targeted dehydration of the cytochrome P450cam (CYP101) distal pocket through mutagenesis of a distal pocket glycine to either valine or threonine results in the alteration of spin state equilibria, and has dramatic consequences on the catalytic rate, coupling efficiency, and kinetic solvent isotope effect parameters, highlighting an important role of the active-site hydration level on P450 catalysis. Cryoradiolysis of the mutant CYP101 oxyferrous complexes further indicates a specific perturbation of proton-transfer events required for the transformation of ferric-peroxo to ferric-hydroperoxo states. Finally, crystallography of the 248Val and 248Thr mutants in both the ferric camphor bound resting state and ferric-cyano adducts shows both the alteration of hydrogen-bonding networks and the alteration of heme geometry parameters. Taken together, these results indicate that the distal pocket microenvironment governs the transformation of reactive heme-oxygen intermediates in P450 cytochromes.     

Structure of a Moloney murine leukemia virus-virus-like 30 recombinant: implications for transduction of the c-Ha-ras proto-oncogene.

Tumor progression locus 2 (Tpl-2) encodes a novel serine-threonine protein kinase which is activated by provirus integration in the late stages of oncogenesis in Moloney leukemia virus (MoMuLV) induced rat T-cell lymphomas. In this report, we present evidence that the provirus integrated in the Tpl-2 locus in 1 of 10 T-cell lymphomas harboring a Tpl-2 rearrangement (2779) is a recombinant between MoMuLV and virus-like 30 (VL30) sequences (Mo-VL30). Recombination between MoMuLV and VL30 may contribute to the transduction of ras, as suggested by the finding that VL30 flanks the ras oncogene in all of the ras transducing viruses isolated from rats to date. The Mo-VL30 recombinant described here represents evidence that recombination between MoMuLV and VL30 can be uncoupled from the transduction of ras, and it may precede the transduction. Sequence comparison between clones of Mo-VL30, Harvey sarcoma virus (Ha-MSV), and genomic c-Ha-ras revealed that all three share a 124-bp region of 87.3% homology. This region was detected at nucleotide positions -1845 to -1720 of c-Ha-ras and 20 bp 5' of the recombination breakpoint between VL30 and ras in Ha-MSV. On the basis of the sequence comparison between VL30, Ha-MSV, and c-Ha-ras, we are proposing a model which explains how VL30 may have facilitated the transduction of c-Ha-ras and perhaps the other ras proto-oncogenes. According to this model, the sequence homology between VL30 and c-Ha-ras targets this gene for transduction by promoting the integration of the provirus in this locus through homologous recombination.     

Utilization of a new index to distinguish heterozygous thalassemic syndromes: comparison of its specificity to five other discriminants

In 1984, a new index (the Makris index) combining erythrocytic and platelet parameters was described for the discrimination of the heterozygous thalassemic syndromes (beta and alpha). The algorithm is [(MCV/RDW)/(MPV/PDW)] divided by the RBC count in millions and requires for input the MCV, RDW, MPV, PDW, and RBC. The critical value used for separating the heterozygous thalassemic subjects is 1.84, which is the mean value plus 2 SD of our heterozygous subjects (confidence limit 95%). Because this index utilizes a confidence limit that includes 95% of affected persons, all individuals with values smaller than this need further investigation. It should be noted that the specificity of the index can be increased using the mean value of our heterozygous group plus 3 SD (X + 3 SD = 1.30 + 3*0.27 = 2.11, confidence limit 99%). In a series of 1510 "normal" subjects examined, 154 were designated as abnormals. None of the rest had abnormalities of cellular morphology or red cells osmotic resistance. The algorithm is readily incorporated into the software of an automated, whole blood analyzer. Using an expert system, we compared the sensitivity and specificity of the Makris index to five other discriminants (Mentzer, Shine et al., England et al., Green, and Bessman et al.). The Makris index distinguished between heterozygotes and normals without misdiagnosis.     

Methods of MRI-based structural imaging in the aging monkey

Rhesus monkeys, whose typical lifespan can be as long as 30 years in the presence of veterinary care, undergo a cognitive decline as a function of age. While cortical neurons are largely preserved in the cerebral cortex, including primary motor and visual cortex as well as prefrontal association cortex there is marked breakdown of axonal myelin and an overall reduction in white matter predominantly in the frontal and temporal lobes. Whether the myelin breakdown is diffuse or specific to individual white matter fiber pathways is important to be known with certainty. To this end the delineation and quantification of specific frontotemporal fiber pathways within the frontal and temporal lobes is essential to determine which structures are altered and the extent to which these alterations correlate with behavioral findings. The capability of studying the living brain non-invasively with MRI opens up a new window in structural–functional and anatomic–clinical relationships allowing the integration of information derived from different scanning modalities in the same subject. For instance, for any particular voxel in the cerebrum we can obtain structural T1-, diffusion- and magnetization transfer- magnetic resonance imaging (MRI) based information. Moreover, it is thus possible to follow any observed changes longitudinally over time. These acquisitions of multidimensional data in the same individual within the same MRI experimental setting would enable the creation of a data base of integrated structural MRI-behavioral correlations for normal aging monkeys to elucidate the underlying neurobiological mechanisms of functional senescence in the aging non-human primate.     

Microstructural abnormalities in subcortical reward circuitry of subjects with major depressive disorder

Background

Previous studies of major depressive disorder (MDD) have focused on abnormalities in the prefrontal cortex and medial temporal regions. There has been little investigation in MDD of midbrain and subcortical regions central to reward/aversion function, such as the ventral tegmental area/substantia nigra (VTA/SN), and medial forebrain bundle (MFB).

Methodology/Principal Findings

We investigated the microstructural integrity of this circuitry using diffusion tensor imaging (DTI) in 22 MDD subjects and compared them with 22 matched healthy control subjects. Fractional anisotropy (FA) values were increased in the right VT and reduced in dorsolateral prefrontal white matter in MDD subjects. Follow-up analysis suggested two distinct subgroups of MDD patients, which exhibited non-overlapping abnormalities in reward/aversion circuitry. The MDD subgroup with abnormal FA values in VT exhibited significantly greater trait anxiety than the subgroup with normal FA values in VT, but the subgroups did not differ in levels of anhedonia, sadness, or overall depression severity.

Conclusions/Significance

These findings suggest that MDD may be associated with abnormal microstructure in brain reward/aversion regions, and that there may be at least two subtypes of microstructural abnormalities which each impact core symptoms of depression.