In conditions of limited chromophore supply rods entrap 11-cis-retinal leading to loss of cone function and cell death.

RPE65 is a retinoid isomerase required for the production of 11-cis-retinal, the chromophore of both cone and rod visual pigments. We recently established an R91W knock-in mouse strain as homologous animal model for patients afflicted by this mutation in RPE65. These mice have impaired vision and can only synthesize minute amounts of 11-cis-retinal. Here, we investigated the consequences of this chromophore insufficiency on cone function and pathophysiology. We found that the R91W mutation caused cone opsin mislocalization and progressive geographic cone atrophy. Remnant visual function was mostly mediated by rods. Ablation of rod opsin corrected the localization of cone opsin and improved cone retinal function. Thus, our analyses indicate that under conditions of limited chromophore supply rods and cones compete for 11-cis-retinal that derives from regeneration pathway(s) which are reliant on RPE65. Due to their higher number and the instability of cone opsin, rods are privileged under this condition while cones suffer chromophore deficiency and degenerate. These findings reinforce the notion that in patients any effective gene therapy with RPE65 needs to target the cone-rich macula directly to locally restore the cones' chromophore supply outside the reach of rods.

Métabolisme d'antidépresseurs dans différentes espèces : aspects méthodologiques et pharmacogénétiques, perspectives cliniques

Résumé pour large public Unité de Biochimie et Psychopharmacologie Clinique, Centre de neurosciences Psychiatrique, Département de Psychiatrie Adulte, Faculté de Biologie et de Médecine, Université de Lausanne Lors de la prise d'un médicament, celui-ci va passer par différentes étapes que sont l'absorption, la distribution, le métabolisme et enfin l'élimination. Ces quatre étapes sont regroupées sous le nom de pharmacocinétique. A noter que ces quatre paramètres sont dynamiques et en constante évolution. Durant cette thèse, nous avons investigué différents aspects de la pharmacocinétique, tout d'abord par une revue de la littérature sur la glycoprotéine-P (Pgp). Récemment découverte, cette protéine de membrane est située aux endroits stratégiques de l'organisme comme la barrière hématoencéphalée, le placenta ou les intestins où elle influencera l'entrée de différentes substances, en particulier les médicaments. La Pgp serait impliquée dans les phénomènes de résistances aux agents thérapeutiques en oncologie. La Pgp influence donc l'absorption des médicaments, et son impact en clinique, en termes d'efficacité de traitement et de toxicité prend chaque jour plus d'importance. Ensuite nous avons mis au point une méthode d'analyse quantitative d'un antidépresseur d'une nouvelle génération : la mirtazapine (Remeron®). La nouveauté réside dans la façon dont la mirtazapine interagit avec les neurotransmetteurs impliqués dans la dépression que sont la sérotonine et la noradrénaline. Cette méthode utilise la chromatographie liquide pour séparer la mirtazapine de ses principaux métabolites dans le sang. La spectrométrie de masse est utilisée pour les détecter et les quantifier. Les métabolites sont des substances issues de réactions chimiques entre la substance mère, la mirtazapine, et généralement des enzymes hépatiques, dans le but de rendre cette substance plus soluble en vue de son élimination. Cette méthode permet de quantifier la mirtazapine et ses métabolites dans le sang de patients traités et de déterminer la variation des taux plasmatiques chez ces patients. Puis nous avons étudié le métabolisme d'un autre antidépresseur, le citalopram, qui a un métabolisme complexe. Le citalopram est un racémate, c'est-à-dire qu'il existe sous forme de deux entités chimiques (R-(-) et S-(+) citalopram) qui ont le même nombre d'éléments mais arrangés différemment dans l'espace. La voie métabolique cérébrale du citalopram est sous le contrôle d'une enzyme, la monoamine oxydase (MAO), conduisant à une forme acide du citalopram (l'acide propionique du citalopram). La MAO existe sous deux formes : MAO-A et MAO-B. Nous avons utilisé des souris déficientes d'un gène, celui de la MAO-A, pour mieux en comprendre le métabolisme en les comparants à des souris sauvages (sans déficience de ce gène). Nous avons utilisé le citalopram et deux de ses métabolites (le déméthylcitaloprarn et le didéméthyícitalopram) comme substrats pour tester la formation in vitro de l'acide propionique du citalopram. Nos résultats montrent que la MAO-A favorise la formation de l'entité R-(-) et présente une plus grande affinité pour le citalopram, tandis que la MAO-B métabolise préférentiellement l'entité S-(+) et a une plus grande affinité pour les deux métabolites déméthylés. De plus, la déficience en MAO-A est partiellement compensée parla MAO-B chez les souris déficientes du gène de la MAO-A. Enfin, nous avons étudié une deuxième voie métabolique du citalopram qui s'est avérée toxique chez le chien Beagle. Celle-ci est catalysée par une autre famille d'enzymes, les cytochromes P-450, et mène aux métabolites déméthylés et didéméthylés du citalopram. Nous avons utilisé des tissus hépatiques de chiens Beagle. Plusieurs cytochromes P-450 sont impliqués dans le métabolisme du citalopram menant à sa forme déméthylée, ceci tant chez l'homme que chez le chien. Par contre, dans le métabolisme de la forme déméthylée menant à 1a forme didéméthylée, un seul cytochrome P-450 serait impliqué chez l'Homme, tandis qu'ils seraient plusieurs chez le chien. L'activité enzymatique produisant la forme didéméthylée est beaucoup plus importante chez le chien comparé à l'homme. Cette observation soutien l'hypothèse que des taux élevés de la forme didéméthylée participent à la toxicité spécifique du citalopram chez le chien. Nous pouvons conclure que plusieurs famille d'enzymes sont impliquées tant au niveau cérébral qu'hépatique dans la métabolisation de médicaments psychotropes. Sachant que les enzymes peuvent être stimulées ou inhibées, il importe de pouvoir suivre au plus prés les taux plasmatiques des différents psychotropes et de leurs métabolites. Résumé Unité de Biochimie et Psychopharmacologie Clinique, Centre de neurosciences Psychiatrique, Département de Psychiatrie Adulte, Faculté de Biologie et de Médecine, Université de Lausanne La plupart des médicaments subissent une transformation enzymatique dans l'organisme. Les substances issues de cette métabolisation ne sont pas toujours dotées d'une activité pharmacologique. Il s'est avéré par conséquent indispensable de suivre les taux plasmatiques d'une substance et de ses métabolites et d'établir ou non l'existence d'une relation avec l'effet clinique observé. Ce concept nommé « therapeutic drag monitoring » (TDM) est particulièrement utile en psychiatrie ou un manque de compliance des patients est fréquemment observé. Les médicaments psychotropes ont un métabolisme principalement hépatique (cytochromes P-450) et parfois cérébral (monoamines oxydases), comme pour le citalopram par exemple. Une méthode stéréosélective de chromatographie liquide couplée à la spectrométrie de masse a été développée pour analyser les énantiomères R-(-) et S-(+) d'un antidépresseur agissant sur les récepteurs noradrénergiques et sérotoninergiques, la mirtazapine et de ses métabolites déméthylmirtazapine et 8-hydroxymirtazapine. Les données préliminaires obtenues dans les plasmas dosés suggèrent que les concentrations de R-(-)-mirtazapine sont plus élevées que celles de S-(+)-mirtazapine, à l'exception des patients qui auraient comme co-médication des inhibiteurs du CYP2D6, telle que la fluoxétine ou la thioridazine. Il y a une enantiosélectivité du métabolisme de la mirtazapine. En particulier pour la 8-hydroxymirtazapine qui est glucuroconjuguée et pour laquelle le ratio S/R varie considérablement. Cette méthode analytique présente l'avantage d'être utilisable pour le dosage stéréosélectif de la mirtazapine et de ses métabolites dans le plasma de patients ayant d'autres substances en co-médication. La glycoprotéine P fonctionne comme une pompe transmembranaire transportant les xénobiotiques depuis le milieu intracellulaire vers le milieu extracellulaire. Son induction et son inhibition, bien que moins étudiées que pour les cytochromes P-450, ont des implications cliniques importantes en termes d'efficacité de traitement et de toxicité. Cette glycoprotéine P a fait l'objet d'une recherche bibliographique. Nous avons étudié le métabolisme du citalopram, un antidépresseur de la classe des inhibiteurs spécifiques de la recapture de la sérotonine chez la souris et chez le chien. Cette substance subit un métabolisme complexe. La voie de métabolisation conduisant à la formation de l'acide propionique du citalopram, catalysée par les monoamines oxydases, a été étudiée in vitro dans les mitochondries cérébrales chez la souris déficiente du gène de la MAO-A (Tg8). La monoamine oxydase A catalyse la formation de l'énantiomère R-(-) et présente une plus grande affinité pour les amines tertiaires, tandis que la monoamine oxydase B favorise la formation de la forme S-(+) et a une affinité plus marquée pour les amines secondaires et primaires. L'étude du citalopram chez la souris Tg8 adulte a montré que la monoamine oxydase B compense la déficience de la monoamine oxydase A chez ces souris génétiquement modifiées. Une autre voie de métabolisation du citalopram conduisant à la formation de didéméthylcitalopram, catalysée par les cytochromes P-450, a été étudiée in vitro dans des microsomes hépatiques de chiens Beagle. Nos études ont montré que les cinétiques de N-déméthylation du citalopram sont biphasiques chez le chien. Les orthologues canins impliqués dans la première N-déméthylation semblent être identiques aux cytochromes P-450 humains. Par contre, dans la deuxième Ndéméthylation, un seul cytochrome P-450 semble être impliqué chez l'homme (CYP2D6), tandis qu'on retrouve jusqu'à cinq orthologues chez le chien. Le CYP2D15, orthologue canin du CYP2D6, est majoritairement impliqué. De plus, l'activité enzymatique, reflétée par les clairances intrinsèques, dans la première N-déméthylation est jusqu'à 45 fois plus élevée chez le chien comparé à l'homme. Ces différentes observations soutiennent l'hypothèse que des taux élevés de didéméthylcitalopram sont responsables de la toxicité du citalopram chez le chien. Nous pouvons conclure que plusieurs famille d'enzymes sont impliquées tant au niveau cérébral qu'hépatique dans la métabolisation de médicaments psychotropes. Sachant -que les enzymes peuvent être induits ou inhibés, il importe de pouvoir suivre au plus près les taux plasmatiques des différents psychotropes et de leurs métabolites. Summary Most of the drugs are metabolized in the organism. Substances issued from this metabolic activity do not always show a pharmacological activity. Therefore, it is necessary to monitor plasmatic levels of drugs and their metabolites, and establish the relationship with the clinical effect. This concept named therapeutic drug monitoring is very useful in psychiatry where lack of compliance is commonly observed. Antidepressants are mainly metabolized in the liver (cytochrome P-450) and sometimes in the brain (monoamine oxidase) like the citalopram, for exemple. A LC-MS method was developed, which allows the simultaneous analysis of R-(-) and S-(+) enantiomers of mirtazapine, an antidepressant acting specifically on noradrenergic and serotonergic receptors, and its metabolites demethylmirtazapine and 8-hydroxymirtazapine in plasma of mirtazapine treated patients. Preliminary data obtained suggested that R-(-) mirtazapine concentrations were higher than those of S-(+) mirtazapine, except in patients comedicated with CYP2D6 inhibitors such as fluoxetine or thioridazine. There is an enantioselectivity in the metabolism of mirtazapine. In particular for the 8-hydroxymirtazapine, which is glucuroconjugated and S/R ratio varies considerably. Therefore this method seems to be suitable for the stereoselective assay of mirtazapine and its metabolites in plasma of patients comedicated with mirtazapine and other drugs for routine and research purposes. P-glycoprotein is working as an efflux transporter of xenobiotics from intracellular to extracellular environment. Its induction or inhibition, although less studied than cytochrome P-450, has huge clinical implications in terms of treatment efficacy and toxicity. An extensive literature search on P-glycoprotein was performed as part of this thesis. The study of citalopram metabolism, an antidepressant belonging to the class of selective serotonin reuptake inhibitors. This substance undergoes a complex metabolism. First metabolization route leading to citalopram propionic acid, catalyzed by monoamine oxidase was studied in vitro in mice brain mitochondria. Monoamine oxidase A catalyzed the formation of R-(-) enantiomer and showed greater affinity for tertiary amines, whereas monoamine oxidase B triggered the formation of S-(+) enantiomer and demonstrated higher affinity for primary and secondary amines. citalopram evaluation in adult Tg8 mice showed that monoamine oxidase B compensated monoamine oxidase A deficiency in those genetically transformed mice. The second metabolization route of citalopram leading to didemethylcitalopram and catalyzed by cytochrome P-450 was studied in vitro in Beagle dog's livers. Our results showed that citalopram N-demethylation kinetics are biphasic in dogs. Canine orthologs involved in the first N-demethylation seemed to be identical to human cytochromes P-450. However, in the second N-demethylation only one cytochrome P-450 seemed to be involved in human (CYP2D6), whereas up to five canine orthologs were found in dogs. CYP2D15 canine ortholog of CYP2D6 was mainly involved. In addition, enzymatic activity reflected by intrinsic clearance in the first N-demethylation was up to 45 fold higher in dogs compared to humans. Those observations support the assumption that elevated rates of didemethylcitalopram are responsible for citalopram toxicity in dogs. We can conclude that several enzymes groups are involved in the brain, as well as in the liver, in antidepressant metabolization. Knowing that enzymes may be induced or inhibited, it makes sense to closely monitor plasmatic levels of antidepressants and their metabolites.

Control and Simulation of Batch Crystallization

Concerning process control of batch cooling crystallization the present work focused on the cooling profile and seeding technique. Secondly, the influence of additives on batch-wise precipitation process was investigated. Moreover, a Computational Fluid Dynamics (CFD) model for simulation of controlled batch cooling crystallization was developed. A novel cooling model to control supersaturation level during batch-wise cooling crystallization was introduced. The crystallization kinetics together with operating conditions, i.e. seed loading, cooling rate and batch time, were taken into account in the model. Especially, the supersaturation- and suspension density- dependent secondary nucleation was included in the model. The interaction between the operating conditions and their influence on the control target, i.e. the constant level of supersaturation, were studied with the aid of a numerical solution for the cooling model. Further, the batch cooling crystallization was simulated with the ideal mixing model and CFD model. The moment transformation of the population balance, together with the mass and heat balances, were solved numerically in the simulation. In order to clarify a relationship betweenthe operating conditions and product sizes, a system chart was developed for anideal mixing condition. The utilization of the system chart to determine the appropriate operating condition to meet a required product size was introduced. With CFD simulation, batch crystallization, operated following a specified coolingmode, was studied in the crystallizers having different geometries and scales. The introduced cooling model and simulation results were verified experimentallyfor potassium dihydrogen phosphate (KDP) and the novelties of the proposed control policies were demonstrated using potassium sulfate by comparing with the published results in the literature. The study on the batch-wise precipitation showed that immiscible additives could promote the agglomeration of a derivative of benzoic acid, which facilitated the filterability of the crystal product.

Is high dose rate brachytherapy reliable and effective treatment for prostate cancer patients? A review of the literature.

The intrinsic physical and radiobiological characteristics of High Dose Rate Brachytherapy (HDR-BT) are well suited to the treatment of prostate cancer. HDR-BT was initially used as a boost to external beam brachytherapy, but has subsequently been employed as the sole treatment, which is termed HDR monotherapy. This review summarizes the clinical outcomes and toxicity results of the principal studies and discusses the radiobiological basis supporting its use.

Mephedrone pharmacokinetics after intravenous and oral administration in rats: relation to pharmacodynamics

Rationale Mephedrone (4-methylmethcathinone) is a still poorly known drug of abuse, alternative to ecstasy or cocaine. Objective The major aims were to investigate the pharmacokineticsa and locomotor activity of mephedrone in rats and provide a pharmacokinetic/pharmacodynamic model. Methods Mephedrone was administered to male Sprague-Dawley rats intravenously (10 mg/kg) and orally (30 and 60 mg/kg). Plasma concentrations and metabolites were characterized using LC/MS and LC-MS/MS fragmentation patterns. Locomotor activity was monitored for 180-240 min. Results Mephedrone plasma concentrations after i.v. administration fit a two-compartment model (α=10.23 h−1, β=1.86 h−1). After oral administration, peak mephedrone concentrations were achieved between 0.5 and 1 h and declined to undetectable levels at 9 h. The absolute bioavailability of mephedrone was about 10 % and the percentage of mephedrone protein binding was 21.59±3.67%. We have identified five phase I metabolites in rat blood after oral administration. The relationship between brain levels and free plasma concentration was 1.85±0.08. Mephedrone induced a dose-dependent increase in locomotor activity, which lasted up to 2 h. The pharmacokinetic-pharmacodynamic model successfully describes the relationship between mephedrone plasma concentrations and its psychostimulant effect. Conclusions We suggest a very important first-pass effect for mephedrone after oral administration and an easy access to the central nervous system. The model described might be useful in the estimation and prediction of the onset, magnitude,and time course of mephedrone pharmacodynamics as well as to design new animal models of mephedrone addiction and toxicity.

Role of brachytherapy in the treatment of cancers of the anal canal. Long-term follow-up and multivariate analysis of a large monocentric retrospective series.

BACKGROUND AND PURPOSE: There are few data on long-term clinical results and tolerance of brachytherapy in anal canal cancer. We present one of the largest retrospective analyses of anal canal cancers treated with external beam radiotherapy with/without (±) chemotherapy followed by a brachytherapy boost. MATERIALS AND METHODS: We performed a retrospective analysis of clinical results in terms of efficacy and toxicity. The impact of different clinical and therapeutic variables on these outcomes was studied. RESULTS: From May 1992 to December 2009, 209 patients received brachytherapy after external beam radiotherapy ± chemotherapy. Of these patients, 163 were stage II or stage IIIA (UICC 2002) and 58 were N1-3. According to age, ECOG performance status (PS), and comorbidities, patients received either radiotherapy alone (58/209) or radiochemotherapy (151/209). The median follow-up was 72.8 months. The 5- and 10-year local control rates were 78.6 and 73.9 %, respectively. Globally, severe acute and late G3-4 reactions (NCI-CTC scale v. 4.0) occurred in 11.2 and 6.3 % of patients, respectively. Univariate analysis showed the statistical impact of the pelvic treatment volume (p = 0.046) and of the total dose (p = 0.02) on the risk of severe acute and late toxicities, respectively. Only six patients required permanent colostomy because of severe late anorectal toxicities. CONCLUSION: After a long follow-up time, brachytherapy showed an acceptable toxicity profile and high local control rates in patients with anal canal cancer.

Mild guanidinoacetate increase under partial guanidinoacetate methyltransferase deficiency strongly affects brain cell development.

Among cerebral creatine deficiency syndromes, guanidinoacetate methyltransferase (GAMT) deficiency can present the most severe symptoms, and is characterized by neurocognitive dysfunction due to creatine deficiency and accumulation of guanidinoacetate in the brain. So far, every patient was found with negligible GAMT activity. However, GAMT deficiency is thought under-diagnosed, in particular due to unforeseen mutations allowing sufficient residual activity avoiding creatine deficiency, but enough guanidinoacetate accumulation to be toxic. With poorly known GAA-specific neuropathological mechanisms, we developed an RNAi-induced partial GAMT deficiency in organotypic rat brain cell cultures. As expected, the 85% decrease of GAMT protein was insufficient to cause creatine deficiency, but generated guanidinoacetate accumulation causing axonal hypersprouting and decrease in natural apoptosis, followed by induction of non-apoptotic cell death. Specific guanidinoacetate-induced effects were completely prevented by creatine co-treatment. We show that guanidinoacetate accumulation without creatine deficiency is sufficient to affect CNS development, and suggest that additional partial GAMT deficiencies, which may not show the classical brain creatine deficiency, may be discovered through guanidinoacetate measurement.

Characterization of nutrient deficiency in Hancornia speciosa Gomes seedlings by omitting micronutrients from the nutrient solution

Hancornia speciosa Gomes (Mangaba tree) is a fruit tree belonging to the Apocynaceae family and is native to Brazil. The production of seedlings of this species is limited by a lack of technical and nutritional expertise. To address this deficiency, this study aimed to characterize the visual symptoms of micronutrient deficiency and to assess growth and leaf nutrient accumulation in H. speciosa seedlings supplied with nutrient solutions that lack individual micronutrients. H. speciosa plants were grown in nutrient solution in a greenhouse according to a randomized block design, with four replicates. The treatments consisted of a group receiving complete nutrient solution and groups treated with a nutrient solution lacking one of the following micronutrients: boron (B), copper (Cu), iron (Fe), manganese (Mn), zinc (Zn), and molybdenum (Mo). The visual symptoms of nutrient deficiency were generally easy to characterize. Dry matter production was affected by the omission of micronutrients, and the treatment lacking Fe most limited the stem length, stem diameter, root length, and number of leaves in H. speciosa seedlings as well as the dry weight of leaves, the total dry weight, and the relative growth in H. speciosa plants. The micronutrient contents of H. speciosa leaves from plants receiving the complete nutrient solution treatment were, in decreasing order, Fe>Mn>Cu>Zn>B.

Proteinuria Increases Plasma Phosphate by Altering Its Tubular Handling.

Proteinuria and hyperphosphatemia are cardiovascular risk factors independent of GFR. We hypothesized that proteinuria induces relative phosphate retention via increased proximal tubule phosphate reabsorption. To test the clinical relevance of this hypothesis, we studied phosphate handling in nephrotic children and patients with CKD. Plasma fibroblast growth factor 23 (FGF-23) concentration, plasma phosphate concentration, and tubular reabsorption of phosphate increased during the proteinuric phase compared with the remission phase in nephrotic children. Cross-sectional analysis of a cohort of 1738 patients with CKD showed that albuminuria≥300 mg/24 hours is predictive of higher phosphate levels, independent of GFR and other confounding factors. Albuminuric patients also displayed higher plasma FGF-23 and parathyroid hormone levels. To understand the molecular mechanisms underlying these observations, we induced glomerular proteinuria in two animal models. Rats with puromycin-aminonucleoside-induced nephrotic proteinuria displayed higher renal protein expression of the sodium-phosphate co-transporter NaPi-IIa, lower renal Klotho protein expression, and decreased phosphorylation of FGF receptor substrate 2α, a major FGF-23 receptor substrate. These findings were confirmed in transgenic mice that develop nephrotic-range proteinuria resulting from podocyte depletion. In vitro, albumin did not directly alter phosphate uptake in cultured proximal tubule OK cells. In conclusion, we show that proteinuria increases plasma phosphate concentration independent of GFR. This effect relies on increased proximal tubule NaPi-IIa expression secondary to decreased FGF-23 biologic activity. Proteinuria induces elevation of both plasma phosphate and FGF-23 concentrations, potentially contributing to cardiovascular disease.

Phenotipic comparison of clinical and plant beneficial strains of Pantoea agglomerans

Certain strains of Pantoea are used as biocontrol agents for the suppression of plant diseases. However, their commercial registration is hampered in some countries because of biosafety concerns. This study compares clinical and plant-beneficial strains of P. agglomerans and related species using a phenotypic analysis approach in which plant-beneficial effects, adverse effects in nematode models, and toxicity were evaluated. Plant-beneficial effects were determined as the inhibition of apple fruit infection by Penicillium expansum and apple flower infection by Erwinia amylovora. Clinical strains had no general inhibitory activity against infection by the fungal or bacterial plant pathogens, as only one clinical strain inhibited P. expansum and three inhibited E. amylovora. By contrast, all biocontrol strains showed activity against at least one of the phytopathogens, and three strains were active against both. The adverse effects in animals were evaluated in the plant-parasitic nematode Meloidogyne javanica and the bacterial-feeding nematode Caenorhabditis elegans. Both models indicated adverse effects of the two clinical strains but not of any of the plant-beneficial strains. Toxicity was evaluated by means of hemolytic activity in blood, and genotoxicity with the Ames test. None of the strains, whether clinical or plant-beneficial, showed any evidence of toxicity

The influence of substratum type and nutrient supply on biofilm organic matter utilization in streams

We investigated the effect of benthic substratum type (sand and rocks) and nutrient supply (N and P) on biofilm structure and heterotrophic metabolism in a field experiment in a forested Mediterranean stream (Fuirosos). Rock and sand colonization and biofilm formation was intensively studied for 44 d at two stream reaches: control and experimental (continuous addition of phosphate, ammonia, and nitrate). Structural (C, N, and polysaccharide content and bacterial and chlorophyll density) and metabolic biofilm parameters (b-glucosidase, peptidase, and phosphatase enzyme activities) were analyzed throughout the colonization process. The epilithic biofilm (grown on rocks) had a higher peptidase activity at the impacted reach, together with a higher algal and bacterial biomass. The positive relationship between the peptidase activity per cell and the N content of the epilithic biofilm suggested that heterotrophic utilization of proteinaceous compounds from within the biofilm was occurring. In contrast, nutrient addition caused the epipsammic biofilm (grown on sand) to exhibit lower b-glucosidase and phosphatase activities, without a significant increase in bacterial and algal biomass. The differential response to nutrient addition was related to different structural characteristics within each biofilm. The epipsammic biofilm had a constant and high C:N ratio (22.7) throughout the colonization. The epilithic biofilm had a higher C:N ratio at the beginning of the colonization (43.2) and evolved toward a more complex structure (high polysaccharide content and low C:N ratio) during later stages. The epipsammic biofilm was a site for the accumulation and degradation of organic matter: polysaccharides and organic phosphorus compounds had higher degradation activities

Synthesis of 1H-benzoxazine-2,4-diones from heterocyclic anhydrides: evaluation of antioxidant and antimicrobial activities

A facile one-step synthesis of 1H-benzoxazine-2,4-diones from heterocyclic anhydrides and TMSA was described. This paper determines their antimicrobial activity against nine human bacterial pathogens by the broth microdilution method; antioxidant activity by DPPH• inactivation and a ferric-reducing power assay; and toxicity by a brine shrimp, Artemia salina, assay. The 1H-benzoxazine-2,4-dione yields were in the range of 57 to 98%. The novel compound 1H-pyrazino[2,3-][1,3]oxazine-2,4-dione 4c showed the highest antioxidant capacity (DPPH 35.4% and FRAP 0.063 µmol TEs/µmol).

Sorption kinetics and intrapaticulate diffusivity of As(III) bioremediation from aqueous solution, using modified and unmodified coconut fiber

The pollution and toxicity problems posed by arsenic in the environment have long been established. Hence, the removal and recovery remedies have been sought, bearing in mind the efficiency, cost effectiveness and environmental friendliness of the methods employed. The sorption kinetics and intraparticulate diffusivity of As (III) bioremediation from aqueous solution using modified and unmodified coconut fiber was investigated. The amount adsorbed increased as time increased, reaching equilibrium at about 60 minutes. The kinetic studies showed that the sorption rates could be described by both pseudo-first order and pseudo-second order process with the later showing a better fit with a value of rate constant of 1.16 x 10-4 min-1 for the three adsorbent types. The mechanism of sorption was found to be particle diffusion controlled. The diffusion and boundary layer effects were also investigation. Therefore, the results show that coconut fiber, both modified and unmodified is an efficient sorbent for the removal of As (III) from industrial effluents with particle diffusion as the predominant mechanism.

First-Line Chemotherapy with Anthracycline and Taxane Combination in Metastatic Breast Cancer. Detection of bone metastases with TRACP 5b

Background: In Finland, breast cancer (BC) is the most common cancer among women, and prostate cancer (PC) that among men. At the metastatic stage both cancers remain essentially incurable. The goals of therapy include palliation of symptoms, improvement or maintenance of quality of life (QoL), delay of disease progression, and prolongation of survival. Balancing between efficacy and toxicity is the major challenge. With increasing costs of new treatments, appropriate use of resources is paramount. When new treatment regimes are introduced into clinical practice a comprehensive assessment of clinical benefit, adverse effects and cost is necessary. Both BC and PC show a predilection to metastasize to bone. Bone metastases cause significant morbidity impairing the patients´ QoL. Diagnosis of bone metastases relies mainly on radiological methods, which however lack optimal sensitivity and specificity. New tools are needed for detection and follow-up of bone metastases. Aims: Anthracyclines and taxanes are effective chemotherapeutic agents in the treatment of metastatic breast cancer (MBC) with different mechanisms of action. Therefore, evaluation of the combination of anthracyclines with taxanes was a justifiable approach in the treatment of MBC patients. We assessed the efficacy, toxicity, cost of treatment and QoL of BC patients treated with first-line chemotherapy for metastatic disease with the combination epirubicin and docetaxel. We also evaluated the diagnostic potential of tartrate-resistant acid phosphatase 5b (TRACP 5b) and carboxyterminal telopeptides of type I collagen (ICTP) in the diagnosis of bone metastases in BC and TRACP 5b in PC patients. Results: The combination of epirubicin and docetaxel was effective in this phase II study, but required individual dose adjustment to avoid neutropenic infections, and the use of growth factors to maintain a feasible dose level. The response rate was 54 % (95 % CI 37-71) and the median overall survival (OS) was 26 months. Of the patients, 87 % were treated for infections. The treatment of adverse events required additional use of health resources mainly due to neutropenic infections, thereby raising direct treatment costs by 20 %. Despite adverse events, the global QoL was not significantly compromised during the treatment. Clinically evident acute cardiac toxicity was not observed. The combination of serum TRACP 5b and ICTP was at least equally sensitive and specific in detection of of bone metastases as commonly used total alkaline phosphatise (tALP) in BC patients. In contrast, TRACP 5b was less specific and sensitive than tALP as a marker of skeletal changes in PC patients. Conclusions: Treatment with epirubicin and docetaxel showed high efficacy in first-line chemotherapy of MBC. The relatively high incidence of neutropenic infections requiring hospitalization increased the treatment costs. Despite adverse events, the global QoL of the patients was not significantly compromised. The combination of TRACP 5b and ICTP showed similar activity as tALP in detecting bone metastases in MBC. In contrast, TRACP 5b was less specific and sensitive than tALP as a marker of skeletal changes in PC.

Fetal alcohol spectrum disorders in Finnish children and adolescents : diagnosis, cognition, behavior, adaptation and brain metabolic alterations

Alcohol consumption during pregnancy can potentially affect the developing fetus in devastating ways, leading to a range of physical, neurological, and behavioral alterations most accurately termed Fetal Alcohol Spectrum Disorders (FASD). Despite the fact that it is a preventable disorder, prenatal alcohol exposure today constitutes a leading cause of intellectual disability in the Western world. In Western countries where prevalence studies have been performed the rates of FASD exceed, for example, autism spectrum disorders, Down’s syndrome and cerebral palsy. In addition to the direct effects of alcohol, children and adolescents with FASD are often exposed to a double burden in life, as their neurological sequelae are accompanied by adverse living surroundings exposing them to further environmental risk. However, children with FASD today remain remarkably underdiagnosed by the health care system. This thesis forms part of a larger multinational research project, The Collaborative Initiative on Fetal Alcohol Spectrum Disorders (the CIFASD), initiated by the National Institute of Alcohol Abuse and Alcoholism (NIAAA) in the U.S.A. The general aim of the present thesis was to examine a cohort of children and adolescents growing up with fetal alcohol-related damage in Finland. The thesis consists of five studies with a broad focus on diagnosis, cognition, behavior, adaptation and brain metabolic alterations in children and adolescents with FASD. The participants consisted of four different groups: one group with histories of prenatal exposure to alcohol, the FASD group; one IQ matched contrast group mostly consisting of children with specific learning disorder (SLD); and two typically-developing control groups (CON1 and CON2). Participants were identified through medical records, random sampling from the Finnish national population registry and email alerts to students. Importantly, the participants in the present studies comprise a group of very carefully clinically characterized children with FASD as the studies were performed in close collaboration with leading experts in the field (Prof. Edward Riley and Prof. Sarah Mattson, Center for Behavioral Teratology, San Diego State University, U.S.A; Prof. Eugene Hoyme, Sanford School of Medicine, University of South Dakota, U.S.A.). In the present thesis, the revised Institute of Medicine diagnostic criteria for FASD were tested on a Finnish population and found to be a reliable tool for differentiating among the subgroups of FASD. A weighted dysmorphology scoring system proved to be a valuable additional adjunct in quantification of growth deficits and dysmorphic features in children with FASD (Study 1). The purpose of Study 2 was to clarify the relationship between alcohol-related dysmorphic features and general cognitive capacity. Results showed a significant correlation between dysmorphic features and cognitive capacity, suggesting that children with more severe growth deficiency and dysmorphic features have more cognitive limitations. This association was, however, only moderate, indicating that physical markers and cognitive capacity not always go hand in hand in individuals with FASD. Behavioral problems in the FASD group proved substantial compared to the typically developing control group. In Study 3 risk and protective factors associated with behavioral problems in the FASD group were explored further focusing on diagnostic and environmental factors. Two groups with elevated risks for behavioral problems emerged: length of time spent in residential care and a low dysmorphology score proved to be the most pervasive risk factor for behavioral problems. The results underscore the clinical importance of appropriate services and care for less visibly alcohol affected children and highlight the need to attend to children with FASD being raised in institutions. With their background of early biological and psychological impairment compounded with less opportunity for a close and continuous caregiver relationship, such children seem to run an especially great risk of adverse life outcomes. Study 4 focused on adaptive abilities such as communication, daily living skills and social skills, in other words skills that are important for gradually enabling an independent life, maintain social relationships and allow the individual to become integrated into society. The results showed that adaptive abilities of children and adolescents growing up with FASD were significantly compromised compared to both typically-developing peers and IQ-matched children with SLD. Clearly different adaptive profiles were revealed where the FASD group performed worse than the SLD group, who in turn performed worse than the CON1 group. Importantly, the SLD group outperformed the FASD group on adaptive behavior in spite of comparable cognitive levels. This is the first study to compare adaptive abilities in a group of children and adolescents with FASD relative to both a contrast group of IQ-matched children with SLD and to a group of typically-developing peers. Finally, in Study 5, through magnetic resonance spectroscopic imaging (MRS) evidence of longstanding neurochemical alterations were observed in adolescents and young adults with FASD related to alcohol exposure in utero 14-20 years earlier. Neurochemical alterations were seen in several brain areas: in frontal and parietal cortices, corpus callosum, thalamus and frontal white matter areas as well as in the cerebellar dentate nucleus. The findings are compatible with neuropsychological findings in FASD. Glial cells seemed to be more affected than neurons. In conclusion, more societal efforts and resources should be focused on recognizing and diagnosing FASD, and supporting subgroups with elevated risk of poor outcome. Without adequate intervention children and adolescents with FASD run a great risk of marginalization and social maladjustment, costly not only to society but also to the lives of the many young people with FASD.