Lentiviral-mediated delivery of mutant huntingtin in the striatum of rats induces a selective neuropathology modulated by polyglutamine repeat size, huntingtin expression levels, and protein length.

A new strategy based on lentiviral-mediated delivery of mutant huntingtin (htt) was used to create a genetic model of Huntington's disease (HD) in rats and to assess the relative contribution of polyglutamine (CAG) repeat size, htt expression levels, and protein length on the onset and specificity of the pathology. Lentiviral vectors coding for the first 171, 853, and 1520 amino acids of wild-type (19 CAG) or mutant htt (44, 66, and 82 CAG) driven by either the phosphoglycerate kinase 1 (PGK) or the cytomegalovirus (CMV) promoters were injected in rat striatum. A progressive pathology characterized by sequential appearance of ubiquitinated htt aggregates, loss of dopamine- and cAMP-regulated phosphoprotein of 32 kDa staining, and cell death was observed over 6 months with mutant htt. Earlier onset and more severe pathology occurred with shorter fragments, longer CAG repeats, and higher expression levels. Interestingly, the aggregates were predominantly located in the nucleus of PGK-htt171-injected rats, whereas they were present in both the nucleus and processes of CMV-htt171-injected animals expressing lower transgene levels. Finally, a selective sparing of interneurons was observed in animals injected with vectors expressing mutant htt. These data demonstrate that lentiviral-mediated expression of mutant htt provides a robust in vivo genetic model for selective neural degeneration that will facilitate future studies on the pathogenesis of cell death and experimental therapeutics for HD

Silencing mutant ataxin-3 rescues motor deficits and neuropathology in machado-joseph disease transgenic mice.

Machado-Joseph disease (MJD) or spinocerebellar ataxia type 3 (SCA3) is an autosomal dominantly-inherited neurodegenerative disorder caused by the over-repetition of a CAG codon in the MJD1 gene. This expansion translates into a polyglutamine tract that confers a toxic gain-of-function to the mutant protein - ataxin-3, leading to neurodegeneration in specific brain regions, with particular severity in the cerebellum. No treatment able to modify the disease progression is available. However, gene silencing by RNA interference has shown promising results. Therefore, in this study we investigated whether lentiviral-mediated allele-specific silencing of the mutant ataxin-3 gene, after disease onset, would rescue the motor behavior deficits and neuropathological features in a severely impaired transgenic mouse model of MJD. For this purpose, we injected lentiviral vectors encoding allele-specific silencing-sequences (shAtx3) into the cerebellum of diseased transgenic mice expressing the targeted C-variant of mutant ataxin-3 present in 70% of MJD patients. This variation permits to discriminate between the wild-type and mutant forms, maintaining the normal function of the wild-type allele and silencing only the mutant form. Quantitative analysis of rotarod performance, footprint and activity patterns revealed significant and robust alleviation of gait, balance (average 3-fold increase of rotarod test time), locomotor and exploratory activity impairments in shAtx3-injected mice, as compared to control ones injected with shGFP. An important improvement of neuropathology was also observed, regarding the number of intranuclear inclusions, calbindin and DARPP-32 immunoreactivity, fluorojade B and Golgi staining and molecular and granular layers thickness. These data demonstrate for the first time the efficacy of gene silencing in blocking the MJD-associated motor-behavior and neuropathological abnormalities after the onset of the disease, supporting the use of this strategy for therapy of MJD

Head And Heck Mucoepidermoid Carcinoma: Clinicopathologic Study Of 173 Cases

Introduction: Mucoepidermoid carcinoma (MEC) is the most common malignant salivary gland tumor, however few studies have been reported in Brazilian populations. Aim: To report clinical and pathologic data from 173 head and neck MEC treated in the Treatment and Research Center at Hospital do Câncer A. C. Camargo in São Paulo. Study design: Clinical randomized. Material and Method: From 1953 to 1997, 173 cases of MEC were found in the medical files of the center. Data were obtained from the patients' records and histological review of all cases. Results: The mean age of the patients was 44 years and 93 (53.8%) were men; parotid glands were affected in 61 cases (35.2%) and intraoral minor salivary glands in 75 (43.4%). TNM revealed 50.3% of the cases in stages I and II, and histological grading revealed 45.2%, 18.5% and 36.3% low-grade, intermediate-grade and high-grade tumors, respectively. Surgical treatment was employed in 80.3% of the cases, with neck dissection in 52 cases (30.1%), and radiotherapy in 73 (42.2%). Local recurrence, regional and distant metastases were found in 12.7%, 9.8% and 9.2% of the patients, respectively; 5-year and 10-year overall survival rates were 70% and 60%, respectively. Conclusions: MEC affected mainly the parotid gland and the palate of adults, without gender preference. Half of the cases were diagnosed at initial clinical stages and 64% of the tumors were low or intermediate-grade lesions. Surgery was the treatment of choice and prognosis was good.685679684Ellis, G.L., Auclair, P.L., Gnepp, D.R., Surgical Pathology of the salivary glands (1991) Major Problems in Pathology Series, 25. , Philadelphia: WB Saunders CompanyEllis, G.L., Auclair, P.L., Tumors of the Salivary Glands (1996) Armed Forces Institute of Pathology. Atlas of Tumor Pathology. 3 rd Series, , Fascicle 17. WashingtonAuclair, P.L., Goode, R.K., Ellis, G.L., Mucoepidermoid carcinoma of intraoral salivary glands (1992) Cancer, 69, pp. 2021-2030Cardoso, W.P., Denardin, O.V., Rapoport, A., Araujo, V.C., Carvalho, M.B., Proliferating cell nuclear antigen expression in mucoepidermoid carcinoma of salivary glands (2000) São Paulo Med J, 118, pp. 69-74Goode, R.K., Auclair, P.L., Ellis, G.L., Mucoepidermoid carcinoma of the major salivary glands: Clinical and histopathologic analysis of 234 cases with evaluation of grading criteria (1998) Cancer, 82, pp. 1217-1224Brandwein, M.S., Ivanov, K., Wallace, D.I., Hille, J.J., Wang, B., Fahmy, A., Bodian, C., Mills, S.E., Mucoepidermoid carcinoma: A clinicopathologic study of 80 patients with special reference to histological grading (2001) Am J Surg Pathol, 25, pp. 835-845Evans, H.L., Mucoepidermoid carcinoma of salivary glands: A study of 69 cases with special attention to histologic grading (1984) Am J Clin Pathol, 81, pp. 696-701Nascimento, A.G., Amaral, A.L.P., Prado, L.A.F., Kligerman, J., Silveira, T.R.P., Mucoepidermoid carcinoma of salivary glands: A clinicopathologic study of 46 cases (1986) Head Neck Surg, 8, pp. 409-417Plambeck, K., Friedrich, R.E., Bahlo, M., Bartel-Friedrich, S., Klapdor, R., TNM staging, histopathological grading, and tumor-associated antigens in patients with a history of mucoepidermoid carcinoma of the salivary glands (1999) Anticancer Res, 19, pp. 2397-2404Spiro, R.H., Huvos, A.G., Berk, R., Strong, E.W., Mucoepidermoid carcinoma of salivary gland origin: A clinicopathologic study of 367 cases (1978) Am J Surg, 136, pp. 461-468Chinellato, L.E.M., Marquez, I.M., Fleury, R.N., Quevedo, F.C., Estudos da prevalência dos tumores de origem epitelial de glândulas salivares em Serviços de Anatomia Patológica das cidades de Bauru e Jaú (Estado de São Paulo, Brasil) (1994) Rev Fac Odontol Bauru, 2, pp. 45-51Franzi, A.S., Carvalho, M.B., Carcinoma mucoepidermóide avançado das glândulas salivares (1997) Rev Bras Cancerol, 43, pp. 273-280Kusama, K., Iwanari, S., Aisaki, K., Wada, M., Ohtani, J., Itoi, K., Hanai, K., Moro, I., Intraoral minor salivary gland tumors: A retrospective study of 129 cases (1997) J Nihon Univ Sch Dent, 39, pp. 128-132Lopes, M.A., Kowalski, L.P., Santos, G.C., Almeida, O.P., A clinicopathologic study of 196 intraoral minor salivary gland tumors (1999) J Oral Pathol Med, 28, pp. 264-267Loyola, A.M., De Araujo, V.C., De Sousa, S.O.M., De Araujo, N.S., Minor salivary gland tumours: A retrospective study of 164 cases in a Brazilian population (1995) Oral Oncol Eur J Cancer, 31 B, pp. 197-201Rapoport, A., De Andrade Sobrinho, J., Brasilino De Carvalho, M., Magrin, J., Fava, A.S., Cancer of the parotid gland (1981) Int Surg, 66, pp. 243-246Rapoport, A., Carvalho, M.B., Fava, A.S., Góis Filho, J.F., Chagas, J.F.S., Kowalski, L.P., Kanda, J.L., Cheuhen, J.A., Diagnóstico e tratamento das neoplasias das glândulas salivares menores: Estudo de 55 casos (1988) Rev Col Bras Cirur, 15, pp. 289-293Regis De Brito Santos, I., Kowalski, L.P., Cavalcante De Araujo, V., Flavia Logullo, A., Magrin, J., Multivariate analysis of risk factors for neck metastasis in surgically treated parotid carcinomas (2001) Arch Otolaryngol Head Neck Surg, 127, pp. 56-60Spiro, R.H., Thaler, H.T., Hicks, W.F., Kher, U.A., Huvos, A.H., Strong, E.W., The importance of clinical staging of minor salivary gland carcinoma (1991) Am J Surg, 162, pp. 330-336Hicks, M.J., El-Naggar, A.K., Flaitz, C.M., Luna, M.A., Batsakis, J.G., Histocytologic grading of mucoepidermoid carcinoma of major salivary glands in prognosis and survival: A clinicopathologic and flow cytometric investigation (1995) Head Neck, 17, pp. 89-95Hicks, J., Flaitz, C., Mucoepidermoid carcinoma of salivary glands in children and adolescents: Assessment of proliferation markers (2000) Oral Oncol, 36, pp. 454-460Ma'aita, J.K., Al-Kalsi, N., Al-Tamimi, S., Wraikat, A., Salivary gland tumors in Jordan: A retrospective study of 221 patients (1999) Croat Med J, 40, pp. 539-54

Activity-regulated cytoskeleton-associated protein controls AMPAR endocytosis through a direct interaction with clathrin-adaptor protein 2

The activity-regulated cytoskeleton-associated (Arc) protein controls synaptic strength by facilitating AMPA receptor (AMPAR) endocytosis. Here we demonstrate that Arc targets AMPAR to be internalized through a direct interaction with the clathrin-adaptor protein 2 (AP-2). We show that Arc overexpression in dissociated hippocampal neurons obtained from C57BL/6 mouse reduces the density of AMPAR GluA1 subunits at the cell surface and reduces the amplitude and rectification of AMPAR-mediated miniature-EPSCs (mEPSCs). Mutations of Arc, that prevent the AP-2 interaction reduce Arc-mediated endocytosis of GluA1 and abolish the reduction in AMPAR-mediated mEPSC amplitude and rectification. Depletion of the AP-2 subunit µ2 blocks the Arc-mediated reduction in mEPSC amplitude, an effect that is restored by reintroducing µ2. The Arc-AP-2 interaction plays an important role in homeostatic synaptic scaling as the Arc-dependent decrease in mEPSC amplitude, induced by a chronic increase in neuronal activity, is inhibited by AP-2 depletion. These data provide a mechanism to explain how activity-dependent expression of Arc decisively controls the fate of AMPAR at the cell surface and modulates synaptic strength, via the direct interaction with the endocytic clathrin adaptor AP-2

Alterations In The Achilles Tendon After Inflammation In Surrounding Tissue

Objective: To analyze the characteristics of the Achilles tendon of rats after induction of localized inflammation in the rat paw. Methods: In our study three groups were used: inflamed group with carrageenan in rat paw (G1); saline group (G2) and control group (G3). After 4 hours the animals were euthanized and the Achilles tendon removed. Results: No significant differences were observed in the analysis of non-collagenous proteins, glycosaminoglycans and hydroxyproline in the groups but a tendency of reduction was verified in G1. As regards the organization of collagen molecules, no differences were observed between groups. With respect to MMPs activity, a stronger presence of the active isoform of MMP-2 in G1 was observed, suggesting that the remodeling was occurring. Conclusion: Thus, we conclude that the inflammatory process in rat paw may affect the remodeling of tendons located near the inflamed site.205266269Maffulli, N., Kader, D., Tendinopathy of tendo Achillis (2002) J Bone Joint Surg Br, 84 (1), pp. 1-8Maffulli, N., Rupture of the Achilles tendon (1999) J Bone Joint Surg Am, 81 (7), pp. 1019-1036Järvinen, T.A., Kannus, P., Maffulli, N., Khan, K.M., Achilles tendon disorders: etiology and epidemiology (2005) Foot Ankle Clin, 10 (2), pp. 255-266Oliveira, F.S., Pinfildi, C.E., Parizoto, N.A., Liebano, R.E., Bossini, P.S., Garcia, E.B., Effect of low level laser therapy (830 nm) with different therapyregimes on the process of tissue repair in partial lesion calcaneous tendon (2009) Lasers Surg Med, 41 (4), pp. 271-276Karousou, E., Ronga, M., Vigetti, D., Passi, A., Maffulli, N., Collagens, proteoglycans, MMP-2, MMP-9 and TIMPs in human Achilles tendon rupture (2008) Clin Orthop Relat Res, 466 (7), pp. 1577-1582Riley, G., Matrix metalloproteinase activities and their relationship with collagen remodelling in tendon pathology (2002) Matrix Biol, 21 (2), pp. 185-195Dario, B.E., Barquilha, G., Marques, R.M., Lesões esportivas: um estudo com atletas de basquetebol Bauruense (2010) Rev Bras Cienc Esporte, 31 (3), pp. 205-215Vieira, C.P., Aro, A.A., Almeida, M.S., de Mello, G.C., Antunes, E., Pimentel, E.R., Effects of acute inflammation induced in the rat paw on the deep digital flexor tendon (2012) Connect Tissue Res, 53 (2), pp. 160-168Tillander, B., Franzén, L.E., Nilsson, E., Norlin, R., Carrageenan-induced subacromial bursitis caused changes in the rat's rotator cuff (2001) J Orthop Res, 19 (3), pp. 441-447Winter, C.A., Risley, E.A., Nuss, G.W., Carrageenin-induced edema in hind paw of the rat as an assay for antiiflammatory drugs (1962) Proc Soc Exp Biol Med, 111, pp. 544-547Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding (1976) Anal Biochem, 72, pp. 248-254Farndale, R.W., Buttle, D.J., Barrett, A.J., Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue (1986) Biochim Biophys Acta, 883 (2), pp. 173-177Marqueti, R.C., Parizotto, N.A., Chriguer, R.S., Perez, S.E., Selistre-de-Araujo, H.S.A., ndrogenic-anabolic steroids associated with mechanical loading inhibit matrix metallopeptidase activity and affect the remodeling of the achilles tendon in rats (2006) Am J Sports Med, 34 (8), pp. 1274-1280Vidal, B.C., Mello, M.L., Supramolecular order following binding of the dichroic birefringent sulfonic dye Ponceau SS to collagen fibers (2005) Biopolymers, 78 (3), pp. 121-128Riley, G., Tendinopathy:from basic science to treatment (2008) Nat Clin Pract Rheumatol, 4 (2), pp. 82-89Szabo, K.A., Ablin, R.J., Singhingh, G., Matrix metalloproteinases and the immune response (2004) Clin Appl Immun Rev, 4, pp. 295-319Chakraborti, S., Mandal, M., Das, S., Mandal, A., Chakraborti, T., Regulation of matrix metalloproteinases: an overview (2003) Mol Cell Biochem, 253 (1-2), pp. 269-285Magra, M., Maffulli, N., Matrix metalloproteases: a role in overuse tendinopathies (2005) Br J Sports Med, 39 (11), pp. 789-791Clutterbuck, A.L., Harris, P., Allaway, D., Mobasheri, A., Matrix metalloproteinases in inflammatory pathologies of the horse (2010) Vet J, 183 (1), pp. 27-38Marsolais, D., Duchesne, E., Côté, C.H., Frenette, J., Inflammatory cells do not decrease the ultimate tensile strength of intact tendons in vivo and in vitro: protective role of mechanical loading (2007) J Appl Physiol, 102 (1), pp. 11-1

Travelling waves for the Gross-Pitaevskii equation II

The purpose of this paper is to provide a rigorous mathematical proof of the existence of travelling wave solutions to the Gross-Pitaevskii equation in dimensions two and three. Our arguments, based on minimization under constraints, yield a full branch of solutions, and extend earlier results, where only a part of the branch was built. In dimension three, we also show that there are no travelling wave solutions of small energy.Comment: Final version accepted for publication in Communications in Mathematical Physics with a few minor corrections and added remark

Fitomassa de adubos verdes e controle de plantas daninhas em diferentes densidades populacionias de leguminosas.

O objetivo deste trabalho foi avaliar a fitomassa de calopogônio, mucuna-preta, mucunarajada,feijão-de-porco, guandu de porte alto, Crotalaria spectabilis e C. breviflora sob diferentes densidades de semeadura (10, 20, 40, 80 e 160 sementes viáveis m-2), e o crescimento de plantas daninhas nessas densidades, em área de tabuleiros costeiros. O experimento foi desenvolvido de maio a agosto de 1996, no Campo Experimental “Antônio Martins”(EMDAGRO/Embrapa-CPATC), em Lagarto, SE. O número de plantas vivas na floração (NPVF) e a matéria seca da parte aérea das leguminosas (MSPA) foram determinados quando, em cada espécie, cerca de 50% das plantas floresceram. Maiores incrementos de MSPA, em resposta ao adensamento populacional, foram observados em C. spectabilis e C. breviflora, seguidas pelo calopogônio, mucuna-preta e mucuna-rajada. Em relação ao feijão-de-porco, a resposta foi negativa, enquanto com o guandu não houve influência. Quanto ao NPVF, as respostas ao adensamento foram lineares e positivas em C. spectabilis, C. breviflora e calopogônio, e quadráticas com ponto de máxima em feijão-de-porco,guandu e mucuna-rajada. Embora nenhum modelo tenha sido ajustado para expressar a relação entre NPVF e adensamento na semeadura de mucuna-preta, a sobrevivência dessa espécie foi reduzida em todas as densidades. Maiores inibições de plantas daninhas ocorreram nas parcelas de mucuna-preta e feijão-de-porco

From Linear to Nonlinear Response in Spin Glasses: Importance of Mean-Field-Theory Predictions

Deviations from spin-glass linear response in a single crystal Cu:Mn 1.5 at % are studied for a wide range of changes in magnetic field, $\Delta H$. Three quantities, the difference $TRM-(MFC-ZFC)$, the effective waiting time, $t_{w}^{eff}$, and the difference $TRM(t_{w})-TRM(t_{w}=0)$ are examined in our analysis. Three regimes of spin-glass behavior are observed as $\Delta H$ increases. Lines in the $(T,\Delta H)$ plane, corresponding to ``weak'' and ``strong'' violations of linear response under a change in magnetic field, are shown to have the same functional form as the de Almeida-Thouless critical line. Our results demonstrate the existence of a fundamental link between static and dynamic properties of spin glasses, predicted by the mean-field theory of aging phenomena.Comment: 9 pages, 10 figure

Modelling storm hydrodynamics on gravel beaches with XBeach-G

The research in this study was funded by the Engineering and Physical Sciences Research Council (EPSRC; EP/H040056/1). The full text is under embargo until 01.09.1

Estímulo no crescimento e na hidrólise de atp em raízes de alface tratadas com humatos de vermicomposto: ii - efeito da fonte de vermicomposto.

Um dos fatores mais limitantes para a produção de vermicomposto é a disponibilidade de esterco. Neste trabalho, foi avaliado o efeito da substituição parcial do esterco por bagaço de cana e por resíduos de leguminosa (Gliricidia sepium) na vermicompostagem sobre a qualidade do vermicomposto e sobre a bioatividade dos humatos, avaliadas por meio da análise do crescimento radicular e da atividade das bombas de H+ isoladas de raízes de alface. A substituição do esterco por bagaço de cana e por resíduos de leguminosas não acarretou prejuízo às características químicas dos vermicompostos. No entanto, os humatos isolados dos diferentes vermicompostos apresentaram características químicas distintas,tais como: acidez e propriedades óticas distintas. Os humatos produzidos a partir de esterco de bovino e da mistura esterco bovino + bagaço proporcionaram maiores estímulos no crescimento radicular das plantas de alface, sendo os mais indicados para uso na forma solúvel. A inclusão de resíduos de leguminosas no processo de vermicompostagem produziu humatos sem efeito sobre o desenvolvimento das raízes de alface