Peripheral Nerve Repair: Multimodal Comparison of the Long-Term Regenerative Potential of Adipose Tissue-Derived Cells in a Biodegradable Conduit.

Tissue engineering is a popular topic in peripheral nerve repair. Combining a nerve conduit with supporting adipose-derived cells could offer an opportunity to prevent time-consuming Schwann cell culture or the use of an autograft with its donor site morbidity and eventually improve clinical outcome. The aim of this study was to provide a broad overview over promising transplantable cells under equal experimental conditions over a long-term period. A 10-mm gap in the sciatic nerve of female Sprague-Dawley rats (7 groups of 7 animals, 8 weeks old) was bridged through a biodegradable fibrin conduit filled with rat adipose-derived stem cells (rASCs), differentiated rASCs (drASCs), human (h)ASCs from the superficial and deep abdominal layer, human stromal vascular fraction (SVF), or rat Schwann cells, respectively. As a control, we resutured a nerve segment as an autograft. Long-term evaluation was carried out after 12 weeks comprising walking track, morphometric, and MRI analyses. The sciatic functional index was calculated. Cross sections of the nerve, proximal, distal, and in between the two sutures, were analyzed for re-/myelination and axon count. Gastrocnemius muscle weights were compared. MRI proved biodegradation of the conduit. Differentiated rat ASCs performed significantly better than undifferentiated rASCs with less muscle atrophy and superior functional results. Superficial hASCs supported regeneration better than deep hASCs, in line with published in vitro data. The best regeneration potential was achieved by the drASC group when compared with other adipose tissue-derived cells. Considering the ease of procedure from harvesting to transplanting, we conclude that comparison of promising cells for nerve regeneration revealed that particularly differentiated ASCs could be a clinically translatable route toward new methods to enhance peripheral nerve repair.

Involvement of long non-coding RNAs in beta cell failure at the onset of type 1 diabetes in NOD mice.

AIMS/HYPOTHESIS: Exposure of pancreatic beta cells to cytokines released by islet-infiltrating immune cells induces alterations in gene expression, leading to impaired insulin secretion and apoptosis in the initial phases of type 1 diabetes. Long non-coding RNAs (lncRNAs) are a new class of transcripts participating in the development of many diseases. As little is known about their role in insulin-secreting cells, this study aimed to evaluate their contribution to beta cell dysfunction. METHODS: The expression of lncRNAs was determined by microarray in the MIN6 beta cell line exposed to proinflammatory cytokines. The changes induced by cytokines were further assessed by real-time PCR in islets of control and NOD mice. The involvement of selected lncRNAs modified by cytokines was assessed after their overexpression in MIN6 cells and primary islet cells. RESULTS: MIN6 cells were found to express a large number of lncRNAs, many of which were modified by cytokine treatment. The changes in the level of selected lncRNAs were confirmed in mouse islets and an increase in these lncRNAs was also seen in prediabetic NOD mice. Overexpression of these lncRNAs in MIN6 and mouse islet cells, either alone or in combination with cytokines, favoured beta cell apoptosis without affecting insulin production or secretion. Furthermore, overexpression of lncRNA-1 promoted nuclear translocation of nuclear factor of κ light polypeptide gene enhancer in B cells 1 (NF-κB). CONCLUSIONS/INTERPRETATION: Our study shows that lncRNAs are modulated during the development of type 1 diabetes in NOD mice, and that their overexpression sensitises beta cells to apoptosis, probably contributing to their failure during the initial phases of the disease.

Cost-effectiveness of Tenofovir / Emtricitabine / Efavirenz versus Zidovudine / Lamivudine / Efavirenz as fist-line regimens for the treatment of HIV-infected patients in low-income settings

The Highly Active Antiretroviral Therapy (HAART) is the combination of at least three antiretroviral compounds. The combination purpose is to reduce the likelihood of drug resistance. However in the long-term the resistance to the first-line combination occurs and leads to treatment failure. Thus, a second-line and even a third-line regimen are recommended in the long run. [...] [P. 5] The two treatment alternatives under comparison: Tenofovir (300 mg) CO-formulated with Emtricitabine (200 mg) and Efavirenz (600 mg) currently known under the brand name Atripla (R) was introduced in July 2006 in the United States market. The excellent safety profile and ease of use make this combination a perfect first-line regimen in low-income settings. Therefore, this treatment option was recommended in WHO 2006 reviewed guidelines. Unfortunately, Tenofovir and Emtricitabine compounds are still costly and not yet widely available. For a matter of simplification this regimen is referred in this report as "the recent" therapy. Initially, we had in mind to consider the most frequently used first-line regimen in low-income countries (Stavudine / Larnivudme / Nevirapine) as a comparator for this economic evaluation. Unfortunately, according to the literature review results (see Annex 3); there was no data available comparing head to head the effectiveness of this regimen with the recent one. Instead, we selected a less frequently but commonly used first-line regimen in low-income countries as a comparator: Zidovudine, Lamivudine, Efavirenz. This combination has extensive experience in durability, safety and toxicity and seems to be an optimal choice for a first-line regimen according to the clinical trial group 384 team. Furthermore, Zidovudine, one of the compounds of this combination is now recommended as one of the preferred NNRTI [Non Nucleoside Reverse Transcriptase Inhibitors] options to be considered by countries instead of Stavudine (the most used NNRTI in limited-income countries). As this combination has been included in the WHO guidelines as a first-line therapy since 2003 when WHO launched the "3 by 5" scaling-up initiative, this combination of drugs is referred in this report as the "old" therapy. Objectives: The primary objective of this economic evaluation is to compare the two first-line HAARTs introduced above, in a low-income setting context. Both of these combinations are recommended by the 2006 WHO guidelines as potential first-line regimens. The secondary objective is to provide a simplified and comprehensible cost-effectiveness modeling tool in order to help policy makers, in resource-limited settings, make decisions about which first-line HAART to fund using the scarce resources available. [P. 6-7]

Long-term controlled GDNF over-expression reduces dopamine transporter activity without affecting tyrosine hydroxylase expression in the rat mesostriatal system.

The dopamine (DA) transporter (DAT) is a plasma membrane glycoprotein expressed in dopaminergic (DA-) cells that takes back DA into presynaptic neurons after its release. DAT dysfunction has been involved in different neuro-psychiatric disorders including Parkinson's disease (PD). On the other hand, numerous studies support that the glial cell line-derived neurotrophic factor (GDNF) has a protective effect on DA-cells. However, studies in rodents show that prolonged GDNF over-expression may cause a tyrosine hydroxylase (TH, the limiting enzyme in DA synthesis) decline. The evidence of TH down-regulation suggests that another player in DA handling, DAT, may also be regulated by prolonged GDNF over-expression, and the possibility that this effect is induced at GDNF expression levels lower than those inducing TH down-regulation. This issue was investigated here using intrastriatal injections of a tetracycline-inducible adeno-associated viral vector expressing human GDNF cDNA (AAV-tetON-GDNF) in rats, and doxycycline (DOX; 0.01, 0.03, 0.5 and 3mg/ml) in the drinking water during 5weeks. We found that 3mg/ml DOX promotes an increase in striatal GDNF expression of 12× basal GDNF levels and both DA uptake decrease and TH down-regulation in its native and Ser40 phosphorylated forms. However, 0.5mg/ml DOX promotes a GDNF expression increase of 3× basal GDNF levels with DA uptake decrease but not TH down-regulation. The use of western-blot under non-reducing conditions, co-immunoprecipitation and in situ proximity ligation assay revealed that the DA uptake decrease is associated with the formation of DAT dimers and an increase in DAT-α-synuclein interactions, without changes in total DAT levels or its compartmental distribution. In conclusion, at appropriate GDNF transduction levels, DA uptake is regulated through DAT protein-protein interactions without interfering with DA synthesis.

The intrinsic mechanisms of softwood fiber damage in brown stock fiber line unit operations Lappeenranta 2010

The effects of pulp processing on softwood fiber properties strongly influence the properties of wet and dry paper webs. Pulp strength delivery studies have provided observations that much of the strength potential of long fibered pulp is lost during brown stock fiber line operations where the pulp is merely washed and transferred to the subsequent processing stages. The objective of this work was to study the intrinsic mechanisms which maycause fiber damage in the different unit operations of modern softwood brown stock processing. The work was conducted by studying the effects of industrial machinery on pulp properties with some actions of unit operations simulated in laboratory scale devices under controlled conditions. An optical imaging system was created and used to study the orientation of fibers in the internal flows during pulp fluidization in mixers and the passage of fibers through the screen openings during screening. The qualitative changes in fibers were evaluated with existing and standardized techniques. The results showed that each process stage has its characteristic effects on fiber properties: Pulp washing and mat formation in displacement washers introduced fiber deformations especially if the fibers entering the stage were intact, but it did not decrease the pulp strength properties. However, storage chests and pulp transfer after displacement washers contributed to strength deterioration. Pulp screening proved to be quite gentle, having the potential of slightly evening out fiber deformations from very deformed pulps and vice versa inflicting a marginal increase in the deformation indices if the fibers were previously intact. Pulp mixing in fluidizing industrial mixers did not have detrimental effects on pulp strength and had the potential of slightly evening out the deformations, provided that the intensity of fluidization was high enough to allow fiber orientation with the flow and that the time of mixing was short. The chemical and mechanical actions of oxygen delignification had two distinct effects on pulp properties: chemical treatment clearly reduced pulp strength with and without mechanical treatment, and the mechanical actions of process machinery introduced more conformability to pulp fibers, but did not clearly contribute to a further decrease in pulp strength. The chemical composition of fibers entering the oxygen stage was also found to affect the susceptibility of fibers to damage during oxygen delignification. Fibers with the smallest content of xylan were found to be more prone to irreversibledeformations accompanied with a lower tensile strength of the pulp. Fibers poor in glucomannan exhibited a lower fiber strength while wet after oxygen delignification as compared to the reference pulp. Pulps with the smallest lignin content on the other hand exhibited improved strength properties as compared to the references.

Siiderin ja long drink -juomien sekoitusprosessin kehittäminen

Diplomityössä perehdytään Hartwall Lahden tuotantolaitoksen juomanvalmistuksen osaston siiderin ja long drink –juomien valmistusprosessin kehittämiseen. Diplomityön tavoitteena oli löytää prosessin ongelma- ja poikkeamakohtia, tutkia mahdollisia hävikkikohteita ja suorittaa laadun osalta ajoseurantamittauksia. Ongelmakohtien pohjalta laadittiin parannusehdotuksia ja niitä toteutettiin jo tämän projektin aikana. Ongelma- ja poikkeamakohtia tutkittiin ajoprosessin aikana. Hävikkitutkimus kohdennettiin aromiliuoksen annostelulinjaan, nestesokerin an-nostelulinjaan ja tuotteiden alku- ja lopputyönnöille painetankkeihin. Painetankkien osalta mahdollisia hävikkikohteita tutkittiin kokeellisen toiminnan avulla ajo-prosessin aikana. Ajoprosessin käynnistyksen jälkeen selvitettiin milloin valmis tuote on painetankkien venttiilimatriisilla. Uudet parametrit alku- ja lopputyönnöille määritettiin tuoteputken, painetankkien pohjaputkien ja venttiilimatriisin tilavuuksien avulla. Hävikkitutkimuksen tuloksena saatiin pienennettyä hävikkiä ja näin ollen syntyi kustannussäästöjä. Hiilidioksidin sitoutuvuutta tutkittiin mittauksien avulla valmistusprosessin aikana. Mittauksien perusteella havaittiin, että hiilidioksidipitoisuus laskee painetankissa. Tämä johtuu valmiin tuotteen lämpötilamuutoksista. Tuotteen lämpötilaa voidaan tasata esimerkiksi levylämmönvaihtimen avulla.

Tissue transglutaminase (TG2) activity regulates osteoblast differentiation and mineralization in the SAOS-2 cell line

Tissue transglutaminase (type II, TG2) has long been postulated to directly promote skeletal matrix calcification and play an important role in ossification. However, limited information is available on the expression, function and modulating mechanism of TG2 during osteoblast differentiation and mineralization. To address these issues, we cultured the well-established human osteosarcoma cell line SAOS-2 with osteo-inductive conditioned medium and set up three time points (culture days 4, 7, and 14) to represent different stages of SAOS-2 differentiation. Osteoblast markers, mineralization, as well as TG2 expression and activity, were then assayed in each stage. Furthermore, we inhibited TG activity with cystamine and then checked SAOS-2 differentiation and mineralization in each stage. The results showed that during the progression of osteoblast differentiation SAOS-2 cells presented significantly high levels of osteocalcin (OC) mRNA, bone morphogenetic protein-2 (BMP-2) and collagen I, significantly high alkaline phosphatase (ALP) activity, and the increased formation of calcified matrix. With the same tendency, TG2 expression and activity were up-regulated. Furthermore, inhibition of TG activity resulted in a significant decrease of OC, collagen I, and BMP-2 mRNA and of ALP activity and mineralization. This study demonstrated that TG2 is involved in osteoblast differentiation and may play a role in the initiation and regulation of the mineralization processes. Moreover, the modulating effects of TG2 on osteoblasts may be related to BMP-2.

Cellular and molecular studies of the effects of a selective COX-2 inhibitor celecoxib in the cardiac cell line H9c2 and their correlation with death mechanisms

Cardiovascular disease is one of the leading causes of death worldwide, and evidence indicates a correlation between the inflammatory process and cardiac dysfunction. Selective inhibitors of cyclooxygenase-2 (COX-2) enzyme are not recommended for long-term use because of potentially severe side effects to the heart. Considering this and the frequent prescribing of commercial celecoxib, the present study analyzed cellular and molecular effects of 1 and 10 µM celecoxib in a cell culture model. After a 24-h incubation, celecoxib reduced cell viability in a dose-dependent manner as also demonstrated in MTT assays. Furthermore, reverse transcription-polymerase chain reaction analysis showed that the drug modulated the expression level of genes related to death pathways, and Western blot analyses demonstrated a modulatory effect of the drug on COX-2 protein levels in cardiac cells. In addition, the results demonstrated a downregulation of prostaglandin E2 production by the cardiac cells incubated with celecoxib, in a dose-specific manner. These results are consistent with the decrease in cell viability and the presence of necrotic processes shown by Fourier transform infrared analysis, suggesting a direct correlation of prostanoids in cellular homeostasis and survival.

The Impact of Acquisitions on Share Pricing and Long Term Performance – Empirical Evidence from the Global Automotive Industry

Automotive industry has faced intense consolidation pressure, which has lead to increasing number of M&As. However, empirical evidence has given controversial results suggesting that most of M&As are value destructive for acquiring companies and for acquiring companies’ shareholders. The objective of this master’s thesis is to examine how acquiring companies’ shareholders react to acquisition announcement and is the reaction in line with the long-term performance. This study uses empirical evidence from automotive industry, which has been characterized as an industry that holds large amount of vertical and horizontal synergies. Transaction data consists of 65 acquisitions made by publicly listed companies between 2008-2010. The short-term impact is tested by applying event study methodology while the long term operative performance is examined with accounting study methodology. The event study results indicate that during the three days after acquisition (t= 0-2), the acquiring firms’ stocks generate an abnormal return of 1.22% on average across all acquisitions. When long term performance is studied it is evident that acquiring companies perform better than the industry median pre- and post-transaction but there is no statistically significant evidence that the performance has increased. The only performance ratio indicating statistically significant decrease is Return on Equity (ROE). On long-term horizontal acquisitions seem to outperform conglomerate ones but otherwise deal characteristics do not have any statistically significant impact.

The same old line?

As long as you've got metal zippers in your garments you're selling an old line. Today's fashions are much more demanding. They need what a Lightning Zephyr can give them. Part of the tag line for an ad aimed at the garment industry.

Letter with the letterhead “Allan Line of Royal Mail Steamships, Hugh and Andrew Allan, agents, Montreal”

Letter with the letterhead “Allan Line of Royal Mail Steamships, Hugh and Andrew Allan, agents, Montreal”. The salutation is “Dear Sir” and it is signed by Andrew Allan. Mr. Allan had enclosed $150 for shooting rights for last season and is asking if there are any rights available for next year, Dec. 16, 1886.

Suivi à long terme de l’efficacité d’un traitement de physiothérapie sur l'incontinence urinaire d'effort persistante en période postnatale

L’incontinence urinaire d’effort (IUE) est une condition fréquente en période postnatale pouvant affecter jusqu’à 77% des femmes. Neuf femmes sur dix souffrant d’IUE trois mois après l’accouchement, vont présenter une IUE cinq ans plus tard. Le traitement en physiothérapie de l’IUE par le biais d’un programme d’exercices de renforcement des muscles du plancher pelvien est reconnu comme étant un traitement de première ligne efficace. Les études ont prouvé l’efficacité de cette approche sur l’IUE persistante à court terme, mais les résultats de deux ECR à long terme n’ont pas démontré un maintien de l’effet de traitement. L’effet d’un programme en physiothérapie de renforcement du plancher pelvien intensif et étroitement supervisé sur l’IUE postnatale persistante avait été évalué lors d’un essai clinique randomisé il y a sept ans. Le but principal de la présente étude était d’évaluer l’effet de ce programme sept ans après la fin des interventions de l’ECR initial. Un objectif secondaire était de comparer l’effet de traitement à long terme entre un groupe ayant fait seulement des exercices de renforcement du plancher pelvien et un groupe ayant fait des exercices de renforcement du plancher pelvien et des abdominaux profonds. Un troisième objectif était d’explorer l’influence de quatre facteurs de risques sur les symptômes d’IUE et la qualité de vie à long terme. Les cinquante-sept femmes ayant complétées l’ECR initial ont été invitées à participer à l’évaluation du suivi sept ans. Vingt et une femmes ont participé à l’évaluation clinique et ont répondu à quatre questionnaires, tandis que dix femmes ont répondu aux questionnaires seulement. L’évaluation clinique incluait un pad test et la dynamométrie du plancher pelvien. La mesure d’effet primaire était un pad test modifié de 20 minutes. Les mesures d’effets secondaires étaient la dynamométrie du plancher pelvien, les symptômes d’IUE mesuré par le questionnaire Urogenital Distress Inventory, la qualité de vie mesurée par le questionnaire Incontinence Impact Questionnaire et la perception de la sévérité de l’IUE mesuré par l’Échelle Visuelle Analogue. De plus, un questionnaire portant sur quatre facteurs de risques soit, la présence de grossesses subséquentes, la v présence de constipation chronique, l’indice de masse corporel et la fréquence des exercices de renforcement du plancher pelvien de l’IUE, venait compléter l’évaluation. Quarante-huit pour-cent (10/21) des participantes étaient continentes selon de pad test. La moyenne d’amélioration entre le résultat pré-traitement et le suivi sept ans était de 26,9 g. (écart-type = 68,0 g.). Il n’y avait pas de différence significative des paramètres musculaires du plancher pelvien entre le pré-traitement, le post-traitement et le suivi sept ans. Les scores du IIQ et du VAS étaient significativement plus bas à sept ans qu’en prétraitement (IIQ : 23,4 vs 15,6, p = 0,007) et (VAS : 6,7 vs 5,1, p = 0,001). Les scores du UDI étaient plus élevés au suivi sept ans (15,6) qu’en pré-traitement (11,3, p = 0,041) et en post-traitement (5,7, p = 0,00). La poursuite des exercices de renforcement du plancher pelvien à domicile était associée à une diminution de 5,7 g. (p = 0,051) des fuites d’urine observées au pad test selon une analyse de régression linéaire. Les limites de cette étude sont ; la taille réduite de l’échantillon et un biais relié au désir de traitement pour les femmes toujours incontinentes. Cependant, les résultats semblent démontrer que l’effet du traitement à long terme d’un programme de renforcement des muscles du plancher pelvien qui est intensif et étroitement supervisé, est maintenu chez environ une femme sur deux. Bien que les symptômes d’IUE tel que mesuré par les pad test et le questionnaire UDI, semblent réapparaître avec le temps, la qualité de vie, telle que mesurée par des questionnaires, est toujours meilleure après sept qu’à l’évaluation initiale. Puisque la poursuite des exercices de renforcement du plancher pelvien est associée à une diminution de la quantité de fuite d’urine au pad test, les participantes devraient être encouragées à poursuivre leurs exercices après la fin d’un programme supervisé. Pour des raisons de logistique la collecte de donnée de ce projet de recherche s’est continuée après la rédaction de ce mémoire. Les résultats finaux sont disponibles auprès de Chantale Dumoulin pht, PhD., professeure agrée à l’Université de Montréal.

Transient high level mammalian reovirus replication in a bat epithelial cell line occurs without cytopathic effect

Mammalian reoviruses exhibit a large host range and infected cells are generally killed; however, most studies examined only a few cell types and host species, and are probably not representative of all possible interactions between virus and host cell. Many questions thus remain concerning the nature of cellular factors that affect viral replication and cell death. In the present work, it was observed that replication of the classical mammalian reovirus serotype 3 Dearing in a bat epithelial cell line, Tb1.Lu, does not result in cell lysis and is rapidly reduced to very low levels. Prior uncoating of virions by chymotrypsin treatment, to generate infectious subviral particles, increased the initial level of infection but without any significant effect on further viral replication or cell survival. Infected cells remain resistant to virus reinfection and secrete an antiviral factor, most likely interferon, that is protective against the unrelated encephalomyocarditis virus. Although, the transformed status of a cell is believed to promote reovirus replication and viral “oncolysis”, resistant Tb1.Lu cells exhibit a classical phenotype of transformed cells by forming colonies in semisolid soft agar medium. Further transduction of Tb.Lu cells with a constitutively-active Ras oncogene does not seem cell growth or reovirus effect on these cells. Infected Tb1.Lu cells can produce low-level of infectious virus for a long time without any apparent effect, although these cells are resistant to reinfection. The results suggest that Tb1.Lu cells can mount an unusual antiviral response. Specific properties of bat cells may thus be in part responsible for the ability of the animals to act as reservoirs for viruses in general and for novel reoviruses in particular. Their peculiar resistance to cell lysis also makes Tb1.Lu cells an attractive model to study the cellular and viral factors that determine the ability of reovirus to replicate and destroy infected cells.

Line of Brillouin-Erbium Fiber Laser Utilizing a Fabry-Perot Cavity in L-band

In this paper, the generation of a multiwavelength laser source from a Brillouin -Erbium fiber laser in the long wavelength band (L-band) region is experimentally demonstrated. The proposed laser system utilizes a Fabrv-Perot cavity formed by fiber-loop mirrors. Twenty-four lines of Brillouin-Stokes with line spacings of 10 GHz are obtained in the L-band region

Estudio de factibilidad Comercilizadora On-line en Colombia

Internet cambio la forma de hacer negocios, modifico la dinámica del comercio y abrió un gran número de nuevas posibilidades para las empresas con iniciativa de crecimiento. Es indudable que Internet influye cada vez más en las actividades de las personas y de las empresas. Con cerca de 2 mil millones de usuarios en el mundo, y casi 20 millones en Colombia, Internet alcanza directamente a la tercera parte de la población. En este contexto, surge este proyecto de investigación con el objetivo de analizar la factibilidad de comercializar productos por internet y contribuir al desarrollo del comercio electrónico. En primera instancia se realiza un proceso de contextualización en el que se describe el desarrollo del comercio electrónico. En seguida y a manera descriptiva, se expone la situación actual de la comercialización de productos por internet en términos económicos, legales, organizacionales y estadísticos. Finalmente, se evalúa y analiza cada uno de los aspectos del modelo de negocio de comercialización online con el objeto de cumplir con los estándares de calidad del comercio electrónico a nivel internacional para generar de tal manera, la satisfacción de los usuarios que realizan compras por internet y por ende incrementos en la frecuencia de compras, logrando abrir las puertas del comercio electrónico para aprovechar todo su potencial y habilitar nuevos canales para vender productos y servicios a usuarios cada vez más sofisticados. Finalmente se realiza un estudio financiero en el que se evalúa la viabilidad del negocio en el ámbito financiero , realizando unas proyecciones a 10 años del flujo de caja con sus respectivos indicadores financieros que me ayuda a medir que tan factible y beneficiosa es el modelo de negocio.