Introduction The COSTIS system is a commercially available target station for the irradiation of solid targets. Up to 3 targets can be provided for irradiation by a slot system. In standard setup the target can be ejected via a pneumatically driven piston system. The target is then allowed to drop down into an open lead container, which can be closed remotely afterwards. The described procedure is well established and reliable. But the concept is limited to low dose targets and environments. The required entering of the cyclotron vault for manual pick up of the container at the cyclotron and the light 18 mm Pb lead shielding of the container itself cause exposure risk for the personnel after long term irradiations with highly activated cyclotron parts and target. The purpose of this work was the design of an alternative for the pickup and the transport of irradiated targets to minimize the radiation dosage of the personnel during manual handling of the COSTIS-lead container. Principle The new designed transport system still uses the software controlled target ejection function of the COSTIS/IBA-system. With ejection the target capsule is allowed to fall into a PTFE-container. To assure a safe target drop into the PTFE container, the gap between the target guiding plate and the PTFE container is smaller than d/2 of the target capsule. After target ejection the PTFE-container can be transferred remotely from target ejection position (1) to the loading station (2) with a target slide. The loading station allows the transfer of the PTFE container remotely into a lead container (60 mm Pb). Now the vault door is used as carrier of the Pb-container. For this purpose a proper fixture for the Pb-container is mounted at the front side of the vault door and via opening the vault door the container is safely transported out of the vault. Outside the container will be finally closed with a lid and transferred to a trolley for further handling. Due to positioning of the container at a certain altitude together with the deep positioning of the target coin inside of the container, the subsequent closing of the container does not cause significant dosage, a more complicated automatic closing system is not mandatory. After replacement of the lead container further transfers can be executed without entering the vault. For this purpose the exchanged Pb-container is placed at the loading station by closing the vault door and a new PTFE-container will be transferred remotely from a magazine onto the target slide, which again can be re-motely positioned at target ejection position. The magazine of PTFE-Containers holds two replacements in accordance with the maximal capacity of the target slot system of the COSTIS station. The remote system of the transport unit uses redundant feedback signals for a reliable and safe operation. Results and Conclusion The newly implemented transport system allows a significant reduction of the radiation dose during pickup and transport of the irradiated solid targets. No entering of the vault is needed after irradiation. The system is highly reliable due to its redundant and straightforward design (2-fold position switches and photoelectric barriers). Due to fixed attachment points in the vault and at the BTL the mobile unit can be easily removed or mounted. The system is maintenance free and all parts easy accessible. For further handling of the targets lead containers were design to fit in the transfer locks of hot cells. The transfer can be carried out directly from the trolley. Container lid and PTFE container are suited for manipulator handling in hot cells.

Introduction A widely distributed commercially available target station for the irradiation of solid targets is the COSTIS system. The system is specified for beams up to 500 W and is equipped with a front side He-cooling and water cooling on the back side. The target itself has a coin shape with a diameter of 24 mm and thickness of 2 mm. This recommends the system for irradiation of thin targets like foils but it is also useable for irradiation of metal and oxide powders. However the irradiation of powders and granulates is limited due to the dimension of the target capsule. A setup of a capped closed target is hardly achievable. The purpose of this work was the modification of the COSTIS target station for the use of thicker target capsules. This shall enable the more easy and safe handling and irradiation of powdery targets and the use of lockable target capsules. Material and Methods The adaption of the COSTIS system for wider targets is easy and fast achievable by the ex-change of the target guiding plate together with the four distance bolts and their bearings. The effort of the replacement of the standard with the modified parts is comparable with COSTIS maintenance including exchange of the window foil and the O-rings. For the target capsule itself different designs were developed and tested. Now various target capsules are available, depending on required energy, handling needs and properties of the target material. Different locking systems can be used, from “click” capsules to screwable systems. Additionally the tightness of the target capsule can be achieved by placement of on O-ring between the lid and capsule body. Results and Conclusion The wider target body allows the capping of the target material. This enables a wide range of applications. One aspect is the nanoparticle research, where radiolabelling is an excellent tool for in situ online investigations. The chosen design of the target capsule allowed the direct activation of TiO2 nanoparticles. Via the nuclear reaction 48Ti(p,n)48V radiolabelled [48V]TiO2 nanoparticles can be obtained. Another example is the use of recoil effects for radiolabelling of nanoparticles. In this case the kinetic energy of the product of the nuclear reaction 7Li(p,n)7Be is used to implant a radioactive tracer in different nanomaterials like Ag0 – nanoparticles and MWCNT (multi wall carbon nano tubes). In general the irradiation of powders and granulates benefits from the modified design that allows the more flexible adaption to experimental needs.

As major effector cells of the innate immune response, macrophages must adeptly migrate from blood to infected tissues. Endothelial transmigration is accomplished by matrix metalloproteinase (MMP)-induced degradation of basement membrane and extracellular matrix components. The classical activation of macrophages with LPS and IFN-γ causes enhanced microtubule stabilization and secretion of MMPs. Macrophages upregulate MMP-9 expression and secretion upon immunological challenge, and require its activity for migration during inflammatory response. However, the dynamics of MMP-9 production and intracellular distribution, as well as the mechanisms responsible for its trafficking, are unknown. Using immunofluorescent imaging, we localized intracellular MMP-9 to small Golgi-derived cytoplasmic vesicles that contain calreticulin and PDI, in activated macrophages. Vesicular organelles of MMP-9 aligned along stable subsets of microtubules and colocalized with the anterograde molecular motor protein, kinesin. We demonstrated a functional contribution of stable MTs in the enhanced trafficking of MMP-9 extracellularly, and showed that heterogeneity exists in macrophage cell populations with respect to MMP-9 production.

As major effector cells of the innate immune response, macrophages must adeptly migrate from blood to infected tissues. Endothelial transmigration is accomplished by matrix metalloproteinase (MMP)-induced degradation of basement membrane and extracellular matrix components. The classical activation of macrophages with LPS and IFN-γ causes enhanced microtubule stabilization and secretion of MMPs. Macrophages upregulate MMP-9 expression and secretion upon immunological challenge, and require its activity for migration during inflammatory response. However, the dynamics of MMP-9 production and intracellular distribution, as well as the mechanisms responsible for its trafficking, are unknown. Using immunofluorescent imaging, we localized intracellular MMP-9 to small Golgi-derived cytoplasmic vesicles that contain calreticulin and PDI, in activated macrophages. Vesicular organelles of MMP-9 aligned along stable subsets of microtubules and colocalized with the anterograde molecular motor protein, kinesin. We demonstrated a functional contribution of stable MTs in the enhanced trafficking of MMP-9 extracellularly, and showed that heterogeneity exists in macrophage cell populations with respect to MMP-9 production.

There can be no doubt that literature or 'belles lettres' is a vehic1e of thought and culture, and that its authors are among the best keys to their age and its attitudes.

Zugl.: Münster (Westfalen), Universiẗat, Diss., 2002.

This study examined the efficacy of continuous nicotinamide (NAM) administration on recovery of function in rats following traumatic brain injury (TBI). TBI was induced via controlled cortical impact (CCI) bilaterally in the prefrontal cortex (+1.5, 0.0 relative to bregma) or sham surgeries were performed. Rats were then treated with either NAM (150 mg/kg/day) or vehicle (saline). Rats were tested behaviorally on the bilateral tactile adhesive removal task, locomotor placing task, novel exploratory behavior and the Morris water maze (MWM). Rats were also assessed histologically by looking at lesion size, GFAP expression (as a measure of active astroctyes) and MMP-9 expression (as a measure of inflammatory response) at time points of 24 and 48 hours and 30 days. The behavioral assessments showed significant improvements in the NAM-treated animals on the bilateral tactile adhesive removal, locomotor placing and MWM. The histological assessments showed significant lesion reduction at 30 days in the NAM-treated group. There were no differences between NAM-treated and vehicle groups on either GFAP or MMP-9 expression. These results indicate that NAM treatment after TBI can significantly improve recovery of function in rats.

Made available in DSpace on 2014-06-11T19:33:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-12-15Bitstream added on 2014-06-13T19:03:54Z : No. of bitstreams: 1 elias_gp_dr_araca.pdf: 1560641 bytes, checksum: 3dad8c2298005cfeb8d0a078880f5e66 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo deste trabalho foi avaliar a atividade das glândulas salivares, a mineralização dental e a participação da metaloproteinase de matriz (MMP-9) nesta mineralização em filhotes de ratas espontaneamente hipertensas (SHR) tratadas ou não com atenolol. Ratas SHR e normotensas Wistar foram tratadas com atenolol (100mg/Kg/dia, via oral) durante os períodos de prenhez e lactação. Os grupos controle receberam o mesmo volume de água sem atenolol. O fluxo salivar, induzido por nitrato de pilocarpina, a concentração de proteínas (método de Lowry), a atividade da amilase (método cinético a 405 nm), o peso das glândulas salivares (parótidas, submandibulares e sublinguais), a microdureza do esmalte e da dentina de incisivos e molares e a expressão da MMP-9 (imonuperoxidase) no tecido dental foram comparados entre filhotes de ratas SHR e Wistar tratadas ou não com atenolol. Os resultados obtidos foram submetidos ao teste estatístico mais adequado, paramétrico (ANOVA ou test t de Student’s) ou não paramétrico (Kruskal-Wallis), sendo consideradas significativas as diferenças quando p<0,05. Filhotes SHR apresentaram menor fluxo salivar e concentração de proteínas do que filhotes Wistar, mas a atividade da amilase não foi diferente entre os grupos. O peso das glândulas salivares foi semelhante entre filhotes SHR e Wistar... / The objective of the present study was analyzed the salivary activity, the dental mineralization and the role of matrix metalloproteinase-9 (MMP-9) on this mineralization, in pups (30 days) of spontaneously hypertensive rats (SHR) treated, or not treated, with atenolol. Female SHR and normotensive Wistar rats were treated during pregnancy and lactation periods with Atenolol 100mg/Kg/day by oral administration. For the control group, the animals received the same water volume without the drug. The salivary flow rate (stimulated by pilocarpine injection), the protein concentration (Lowry method), salivary amylase activity (kinetic method at 405 nm), the weight of salivary glands (parotid, submandibular and sublingual), the enamel and dentin microhardness of incisors and molars teeth and the matrix metalloproteinase-9 (MMP-9, gelatinase B) localization (imunoperoxidase) in dental tissue were compared between SHR and Wistar pups of female rats treated or not with atenolol. The results were analyzed by parametric (ANOVA or Student s tests) or non-parametric (Kruskal-Wallis) tests (p<0,05). The salivary flow rate and salivary protein concentration were reduced in SHR pups. There was no alteration in amylase activity between groups. The salivary glands weight was not different between SHR and Wistar pups either. Decreased enamel and dentin microhardness were observed in incisors and molar teeth of SHR pups. No alterations in MMP-9 positive staining were observed in predentin and odontoblasts of both groups, however the density of stained ameloblasts cells and external enamel surface were higher in incisors teeth of SHR pups. Atenolol-treated SHR and Wistar rats pups showed decrease in submandibular gland weight, in saliva s flow rate and protein concentration, but no alteration in amylase activity. Atenolol increased enamel and dentin microhardness of incisors teeth of SHR and...(Complete abstract, click electronic address below)

O receptor NLRP12 é um receptor intracelular que está envolvido no reconhecimento de PAMPs e DAMPs durante uma infecção. Foi visto que durante a osteoclastogênese, há uma diminuição da transcrição de NLRP12, e que este receptor inibe a reabsorção óssea através da supressão da via alternativa de NF-&kappa;B, exercendo um papel protetor sobre o tecido ósseo. Além disso, foi observado que a deficiência de NLRP12 em monócitos sob o estímulo de RANKL levou a maior estabilização de NIK e maior translocação de RelB para o núcleo, aumentando a formação dos osteoclastos in vitro. Portanto, o objetivo deste estudo foi avaliar o papel de NLRP12 no desenvolvimento e progressão da lesão periapical induzida em camundongos. Para isso, foi induzida a lesão periapical dos primeiros molares inferiores dos camundongos fêmeas C57/Bl6 (WT) e camundongos deficientes para o receptor NLRP12 (Nlrp12-/-). Após 14 e 21 dias, as amostras de mandíbula foram submetidas às análises: determinação da área de lesão periapical em cortes histológicos; expressão gênica de marcadores osteoclastogênicos por qPCR; contagem de osteoclastos submetidos ao ensaio de histoenzimologia (TRAP); avaliação enzimática das MMPs por Zimografia. Os linfonodos cervicais foram submetidos à análise da expressão dos fatores de transcrição T-bet, ROR&gamma;t, GATA-3 e Foxp-3 por qPCR. Aos 14 dias, na análise histomorfométrica os animais Nlrp12-/- apresentaram uma maior lesão periapical quando comparados aos animais WT, associado com o aumento da expressão de Trap, Catepsina K e MMP-9 em amostras de hemi-mandíbulas com lesão. Além disso, o número de células TRAP positivas foi significantemente maior em Nlrp12-/- com lesão quando comparado com seu controle, enquanto no grupo WT com e sem lesão foram semelhantes. Assim como foi observado o maior aumento dos osteoclastos presentes no local da lesão dos animais Nlrp12-/-, estes também se mostraram com maior atividade gelatinolítica de MMP-9 e MMP-2 em relação ao seu controle, diferente dos animais WT que não apresentaram diferença entre os grupos controle e com lesão. Ainda nesse período, foi observado nas amostras de linfonodos cervicais que, no grupo Nlrp12-/- houve uma tendência à maior expressão de ROR&gamma;t, seguidos de menor expressão de T-bet quando comparados com o grupo WT. Aos 21 dias, os animais WT e Nlrp12-/- apresentaram lesões periapicais de tamanhos semelhantes. Além disso, somente o grupo Nlrp12-/- com lesão apresentou um aumento significativo da expressão de Trap em relação ao seu controle e a expressão de Catepsina K foi semelhante em ambos os grupos. Neste período houve um aumento na quantidade de células TRAP positivas em ambos os grupos com lesão quando comparados com seus respectivos controles, entretanto também não houve diferença entre os animais WT e Nlrp12-/-. A atividade de MMP-9 e MMP-2 foram semelhantes entre os animais WT e Nlrp12-/- e entre seus respectivos controles aos 21 dias. Nossos dados sugerem que a deficiência de NLRP12 levou a uma maior perda óssea aos 14 dias de lesão periapical e que isso ocorre via modulação da formação e atividade dos osteoclastos. Portanto, o NLRP12 inibe a osteoclastogênese e a atividade dos osteoclastos durante a fase inicial da lesão periapical, retardando o desenvolvimento da doença. / NLRP12 is an intracellular sensor that recongnizes PAMPS and DAMPs during infeccion. It was seen that NLRP12 transcription is down-regulated during osteoclastogenesis, and NLRP12 protect bone via suppression of alternative NF-&kappa;B-induced osteoclastogenesis. It has been shown that NLRP12 deficiency in monocytes under RANKL stimuli exhibited more stabilization of NIK and nuclear translocation of RelB, increasing osteoclasts formation in vitro. Therefore, the aim of this study was to evaluate the role of NLRP12 in the development and progression of experimentally induced periapical lesions in mice. Periapical lesions were induced in first molars of WT and NLRP12 knockout (KO) mice. Samples were collected at 14 and 21 days of the lesion for the analyses. Jaw samples with lesion and control area were subjected to periapical lesions area determination by histological sections; osteoclastogenic markers expression by q-PCR; count of osteoclasts submitted to the enzyme assay for TRAP; evaluation of MMPs activity by Zymography. The expression of T cells markers´ transcription factors was evaluated in lymph nodes by q-PCR. Fourteen days after periapical lesion induction, histological analysis revealed that NLRP12KO mice exhibited higher area of periapical lesion compared to WT group, which was associated with up-regulated mRNA expression of Trap, Cathepsin K and MMP-9 in the jaw samples. Moreover, the number of multinuclear TRAP-stained cells was significantly higher in the NLRP12KO with lesion group when compared to it control, whereas in the WT the number between the lesion and control groups were similar. Still, NLRP12KO showed higher MMP-9 and -2 gelatinolytic activity than it control, unlike WT mice that showed no difference between control and lesion group. In this period, NLRP12KO mice lymph nodes showed more ROR&gamma;t expression than WT mice and less T-bet expression. At 21 days, WT and NLRP12KO presented periapical lesions of similar sizes. In addition, NLRP12KO group with lesion showed a significant increase in Trap expression when compared with their control, but the increase in Trap e Cathepsin K was similar in both groups. Additionally, there was an increase of multinuclear TRAP-stained cells in both lesion groups when compared with their respective controls; however, there was also no difference between WT and NLRP12KO mice. MMP-9 and -2 activity was similar between WT and NLRP12KO and with their respective controls at 21 days. Our results suggest that NLRP12 deficiency led to increased bone loss at 14 days of periapical lesion and it occurs due to increased osteoclasts formation and activity. Therefore, NLRP12 inhibits osteoclastogenesis and osteoclasts activity during the early stages of periapical lesion, slowing the development of the disease.

A necessidade de dispositivos analíticos para detecção precoce de câncer tem motivado pesquisas em nanomateriais de baixo custo, com busca de sinergia para obter alta sensibilidade e seletividade em biossensores. Neste trabalho, o ácido 11-mercaptoundecanóico, o polímero natural quitosana e a proteína concanavalina A (Con A) foram usados como plataforma para imobilizar anticorpos anti-CA 19-9 e construir biossensores para detecção de câncer de pâncreas. Esses biossensores foram produzidos com uma matriz de filmes automontados por adsorção química (self-assembled monolayers, SAM) e por adsorção física (layer-by-layer, LbL). A caracterização com técnicas espectroscópicas e gravimétricas permitiu selecionar as arquiteturas com quitosana/ Con A 2:1 (sensor A), quitosana/ Con A 1:1 (sensor B) e 11-MUA (sensor E), como sendo mais favoráveis à imobilização do anticorpo anti-CA 19-9. Usando os biossensores com espectroscopia de impedância foi possível detectar baixas concentrações do biomarcador CA 19-9, com limites de detecção entre 0,17-0,69 U/mL, 0,31-0,91 U/mL e 0,56-0,91 U/mL para os sensores A, B e E, respectivamente. Esses limites são suficientes para detectar câncer de pâncreas nos estágios iniciais. A seletividade dos dispositivos foi inferida em uma série de experimentos de controle com amostras de células SW 620 e HT-29, ácido úrico, ácido ascórbico, glicose, manose, sérum e antígeno p24, indicando ausência de interferência não específica ao biomarcador. O uso de técnicas de visualização de informação permitiu facilmente distinguir essas amostras, classificando-as de acordo com a concentração do biomarcador em um mapa. Permitiu também quantificar a seletividade dos biossensores através do coeficiente de silhueta, com valores 0,853, 0,861 e 0,897 para os sensores A, B e E, respectivamente. Essa especificidade dos biossensores foi confirmada por medidas de PM-IRRAS, através das bandas de amida I e II em 1566 cm-1 e 1650 cm-1, indicando a interação específica entre anticorpo e antígeno, que pode ser modelada com uma isoterma de Langmuir-Freundlich. Quando a matriz de quitosana/ Con A foi substituída por um filme monocamada ou quando se empregou um biomarcador de maior peso molecular, a adsorção do biomarcador foi explicada por uma combinação de dois processos de Langmuir-Freundlich. Conclui-se que os biossensores de baixo custo podem ser eficientes para diagnóstico e prognóstico, e serem implementados na rede nacional de saúde com disseminação da tecnologia. / The need of analytical devices to detect cancer at early stages has motivated research into nanomaterials where synergy is sought to achieve high sensitivity and selectivity in biosensors. In this work, 11-mercaptoundecanoic acid, the polymer chitosan and the protein concanavalin A (Con A) were used as a platform to immobilize the anti-CA 19-9 antibody using the self-assembled monolayer (SAM) and the layer-by-layer (LbL) techniques. The characterization with spectroscopic and gravimetric techniques allowed us to select the architectures with chitosan/Con A 2:1 (Sensor A), chitosan/Con A 1:1 (Sensor B) and 11 MUA (Sensor E) as optimized for immobilization of anti-CA 19-9 antibodies. Using impedance spectroscopy, the biosensors were capable of detecting low concentrations of CA 19-9 biomarker, with limit of detection in the range 0.17-0.69 U/mL, 0.31-0.91 U/mL and 0.56-0.91 U/mL for sensors A, B and E, respectively. These limits are sufficient to detect pancreatic cancer at early stages. The selectivity of the biosensors was inferred in a series of control experiments with cell samples SW-620 and HT-29, uric acid, ascorbic acid, glucose, mannose, serum and p24 antigen, indicating the absence of non-specific interference. With information visualization techniques, these samples could be easily distinguished in a visual map, and be classified according to their content of CA 19-9. Furthermore, the selectivity could be quantified through the silhouette coefficient, with values 0.853, 0.861 and 0.897 for sensors A, B and E, respectively. This biosensor specificity was confirmed with PM-IRRAS measurements by monitoring the amide I and II bands at 1566 cm-1 and 1650 cm-1. The specific interaction between antibody and antigen was modeled with a Langmuir-Freundlich isotherm. When the chitosan/Con A matrix was replaced by a SAM monolayer or if a larger biomarker was employed, adsorption was explained by a combination of two Langmuir-Freundlich processes. In conclusion, low cost biosensors may be effective for diagnostics and prognostics, and may be further implemented in the Brazilian national health system with technology transfer.