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ABSTRACT: The current study compared the effects of intratracheal administration of different lipopolymeric vectors on lung function and histology in normal mice.
Forty-eight BALB/c mice were randomly divided into 8 groups (6/group). All animals received intratracheal instillation of the following suspensions: polymerized [(A) 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC):1,2-bis-(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine (DC8,9PC):1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), (B) DMPC:DC8,9PC:stearylamine (SA), (C) DMPC:DC8,9PC:myristoylcholine chloride (MCl)]; nonpolymerized [(D) DMPC:DC8,9PC:DOTAP, (E) DMPC:DC8,9PC:SA, (F) DMPC:DC8,9PC:MCl] together with plasmid DNA; vehicle (control), and pDsRed2-N1 plasmid DNA (DNA). At 24 h, the survival rate, lung mechanics (resistive and viscoelastic pressure, static elastance) and morphometry were analyzed.
The survival rate was 50% in D, 40% in E and F, and 100% in the CTRL, DNA, A, B and C groups. Animals from groups D, E, and F that died presented diffuse pulmonary hemorrhagic capillaritis. Lung mechanics, the fraction of normal and collapsed alveoli, as well as the number of polymorphonuclear and mononuclear cells in lung tissue were similar in all surviving mice.
Intratracheal instillation of polymerized particles is safe compared with nonpolymerized formulations and may be used for future gene/drug therapy.
Cellular Physiology and Biochemistry 01/2012; 29(5-6):791-8. · 2.86 Impact Factor
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Roberta M Lassance,
Caroline P Pássaro, Sabrina V Martini,
Raquel C Castiglione,
Tatiana M Gutierrez,
Soraia C Abreu,
Mariana A Antunes,
Debora G Xisto,
Liudmila Cebotaru,
Hilda Petrs-Silva,
Walter A Zin,
William B Guggino,
Rafael Linden,
Patricia R M Rocco,
Marcelo M Morales
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ABSTRACT: Forty-eight BALB/c mice were divided into two groups of 24 animals each. In the control group (CTRL) saline was intratracheally instilled, while the virus group (VR) received rAAV2-GFP (4 x 10(9) particles). These groups were subdivided into four sub-groups (n=6). Pulmonary mechanical parameters were analyzed after 3 weeks (VR1d3w) and at 1 (VR2d1w), 2 (VR2d2w) and 3 weeks (VR2d3w) after a second AAV2 dose. Fractions of the area of alveolar collapse and the amount of polymorpho- and mononuclear cells were determined by point-counting technique. Viral transduction was evaluated by immunohistochemistry. Lung mechanical data were similar in all groups. However, there was an increase in airway and lung parenchyma cellularity and in the fraction of area of alveolar collapse in the VR2d2w group, which nonetheless decreased with time. There was no evidence of apoptosis in any group. In conclusion, the gene transfer vector AAV2 induces, in the lung, a discrete inflammatory reaction that does not affect either baseline lung mechanics or airway hyperresponsiveness.
Respiratory Physiology & Neurobiology 02/2008; 160(1):91-8. · 2.24 Impact Factor
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ABSTRACT: Rhodnius prolixus Malpighian tubules (MTs) are a good model for fluid and ion secretion studies in view of the dramatic postprandial diuresis, which follows its massive blood meals. Ingestion of a blood meal equals to 10-12 times their initial body mass, leads to rapid activation of high output by excretory system, which eliminates 40-50% of the fluid mass. Secretion of ions and water is stimulated 1000-fold by serotonin and diuretic hormone. These hormones cooperate synergistically to activate adenylate cyclase activity from MTs cells, which increase the level of intracellular cAMP. The anti-diuretic hormones have also an important role in the fluid maintenance of Rhodnius prolixus. Several hours after insect feeding occurs a reduction in urine flow, that has been thought to result from a decreased diuretic hormone release or from a novel mechanism of anti-diuresis involving insect cardioacceleratory peptide 2b (CAP2b) and cyclic GMP. In this article it is discussed how the hormone regulation of fluid transport is done in Rhodnius prolixus MTs.
Anais da Academia Brasileira de Ciências 03/2007; 79(1):87-95. · 1.09 Impact Factor
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ABSTRACT: Thirty-five patients (23 males and 12 females), age 35 +/- 13 years old, presenting either idiopathic calcium nephrolithiasis, nephrocalcinosis or mild renal failure with idiopathic calcium nephrolithiasis were selected for the analysis of low molecular weight proteinuria and the possible mutations occurrence in the chloride channel gene CLCN5. The urinary ratio of beta2-microglobulin and creatinine (beta2M/Cr) was very high in a transplanted woman with nephrocalcinosis (> 3.23 mg/mmol) and slightly high in five patients (> 0.052 or < 1.0 mg/mmol) with multiple urological manipulations. Other studied patients showed beta2M/Cr ratio at normal range (0.003-0.052 mg/mmol) without gender difference (p > 0.05). Mutation analysis of CLCN5 gene was performed in 26 patients of 35 selected (11 with idiopathic hypercalciuria; 6 men with normal calciuria; 3 with mild renal insufficiency and 6 with nephrocalcinosis) and was normal in all subjects even in those with abnormal molecular weight proteinuria. Conclusion: CLCN5 gene mutation is not a common cause of kidney stone disease or nephrocalcinosis in a group of Brazilian patients studied.
Anais da Academia Brasileira de Ciências 03/2005; 77(1):95-101. · 1.09 Impact Factor
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ABSTRACT: The purpose of the present study was to detect the presence of an aquaporin-like water channel, a member of the major intrinsic protein (MIP) family, in the Malpighian tubule (MT) of the hematophagous insect Rhodnius prolixus, which excrete a large bulk of fluid after a massive blood meal, and its possible regulation by 5-hydroxytryptamine (5-HT). Reverse transcription polymerase chain reaction (RT-PCR) and Southern blots of cDNA was obtained from adult R. prolixus MT poly (A)+ RNA. Employing degenerate primers corresponding to the NPA (amino acid sequence motifs repeats Asn-Pro-Ala) highly conserved domain of amino acids sequences of all members of the MIP gene family, we were able to identify a 365-base pair PCR product. The R. prolixus MT mRNA expression of this water transporter is increased in the animal after blood meal and in tubules treated with 5-hydroxytryptamine or cAMP. The up-regulated expression of MT MIP mRNA after a blood meal is probably due to the action of 5-hydroxytryptamine via a cyclic AMP dependent pathway.
Archives of Insect Biochemistry and Physiology 12/2004; 57(3):133-41. · 1.36 Impact Factor
