Chondroitin sulfate proteoglycans are structural renewable constituents of the rabbit vitreous body

Purpose: To characterize the vitreous intrinsic proteoglycans, investigate their dynamics, and examine their role in the supramolecular organization of the vitreous. Methods: Vitreous from normal rabbits was collected and processed for observation with the transmission electron microscope after treatment with glycosidases. Also, rabbits were injected intravitreally with [S-35]-sodium sulfate and sacrificed at several time intervals after the injection. Proteoglycans (PGs) were assayed in the vitreous supernatant or in whole samples extracted with guanidine hydrochloride by polyacrylamide or agarose gel electrophoresis, followed respectively by fluorography or autoradiography, and ion-exchange chromatography and gel-filtration chromatography, combined with glycolytic treatment of the samples. The sulfated glycosaminoglycans (GAGs) were characterized by agarose gel electrophoresis after treating vitreous samples with protease and specific glycosidases. Results: the electron microscopic study revealed a network with hyaluronic acid ( HA) as thin threads coating and connecting collagen fibrils. The elimination of the HA coat showed chondroitin sulfate granules (8-25 nm) arranged at regular intervals on the fibril surface. The chondroitinase ABC digestion, besides removing the granules, also caused the formation of thicker bundles of the collagen fibrils. The PG and GAG analysis indicated that there are three renewable PGs in the vitreous ( e. g., one heparan-and two chondroitin-sulfate ones). Conclusions: At least one of the chondroitin sulfate PGs is involved in the interactions that occur in the vitreous structure, mainly by providing adequate spacing between the collagen fibrils, a condition that is probably required for the transparency of the vitreous.

Influence of ionic interactions on the photoinduced Birefringence of poly[1-[4-(3-carboxy-4 hydroxyphenylazo) benzene sulfonamido]-1 2-ethanediyl, sodium salt] films

Electrostatic interactions govern most properties of polyelectrolyte films, as in the photoinduced bire-fringence of azo-containing polymers. In this paper we report a systematic investigation of optical storage characteristics of cast and layer-by-layer (LbL) films of poly[1 -[4-(3-carboxy-4 hydroxypheny-lazo) benzene sulfonamido]-1,2-ethanediyl, sodium salt] (PAZO). Birefringence was photoinduced faster in PAZO cast films prepared at high pHs, with the characteristic writing times decreasing almost linearly with the pH in the range between 4 and 9. This was attributed to an increased free volume for the azochromophores with the enhanced electrostatic repulsion in PAZO charged to a greater extent. In contrast, in LbL films of PAZO alternated with poly(allylamine hydrochloride) (PAH), the electrostatic interactions between the oppositely charged polymers hampered photoisomerization and molecular rearrangement, thus leading to a slower writing kinetics for highly charged PAH or PAZO.

LATERAL PARABRACHIAL SEROTONERGIC MECHANISMS - ANGIOTENSIN-INDUCED PRESSOR AND DRINKING RESPONSES

This study investigated the effects of bilateral injections of serotonergic receptor ligands into the lateral parabrachial nucleus (LPBN) on the presser and dipsogenic responses induced by intracerebroventricular (icv) injection of angiotensin II (ANG II). Rats with stainless steel cannulas implanted bilaterally into the LPBN and into the left lateral ventricle were used to study icy ANG II-induced water intake and presser responses. Pretreatment with the serotonergic 5-HT1/5-HT2 receptor antagonist methysergide (1-8 mu g/200 nl) bilaterally injected into the LPBN increased the water intake induced by icv ANG II (50 ng/mu l) administered via the lateral ventricle, but pretreatment with methysergide (4 mu g/200 nl) did not change the presser response produced by icy ANG II. After bilateral injection of either serotonin (5-HT, 5 mu g/200 nl) or the serotonergic 5-HT2a/5-HT2c receptor agonist (+/-)-2,5-dimetoxy-4-iodoamphetamine hydrochloride (DOI; 0.5-10 mu g/200 nl) into the LPBN, the water intake induced by ANG II was significantly reduced. These results are consistent with other observations indicating that the LPBN is associated with inhibitory mechanisms controlling water intake induced by ANG II treatment and suggest that serotonergic pathways may be involved in this effect.

Morphology characterization of layer-by-layer films from PAH/MA-co-DR13: the role of film thickness

We report on the use of dynamic scale theory and fractal analyses in the Study of distinct growth stages of layer-by-layer (LBL) films of poly(allylamine hydrochloride) (PAH) and a side-chain-substituted azobenzene copolymer (Ma-co-DR13). The LBL films were adsorbed oil glass substrates and characterized with atomic force microscopy with the Ma-co-DR13 at the top layer. The ganular morphology exhibited by the films allowed the observation of the growth process inside and outside the grains. The growth outside the grains was found to follow the Kardar-Parisi-Zhang model, with fractal dimensions of ca. 2.6. One could expect that inside the grains the morphology would be close to a Euclidian surface with fractal dimension of ca. 2 for any growth stage. The latter, however, was observed only for thicker films containing more than 10 bilayers. For thinner films the morphology was well described by a self-affine fractal. Such dependence of the growth behavior with the film thickness is associated with a more complete coverage of adsorption sites in thicker films due to diffusion of polymer molecules. (c) 2004 Elsevier B.V. All rights reserved.

Healing of dehiscence defects following root surface demineralization with tetracycline: A histologic study in monkeys

Background: the purpose of this study was to histologically evaluate the healing of experimental dehiscence defects after surface demineralization with tetracycline hydrochloride.Methods: Six adult male monkeys (Cebus apella) were used in this study. Dehiscence defects were surgically created on the buccal aspect of the mandibular lateral incisors in all animals. The root surfaces were debrided and planed. In a split-mouth design, a 10% tetracycline hydrochloride solution was applied to one tooth for 4 minutes (T group), followed by irrigation with saline. The contralateral tooth served as a control (C group). The flaps were repositioned and sutured. The animals were sacrificed at 6 months postoperatively and histological sections were processed. Computer-assisted histomorphometric analysis was used to evaluate the formation of new cementum, new bone, new connective tissue attachment, and length of the epithelium (junctional and sulcular).Results: Bone regeneration was similar in both groups (1.5 +/- 0.3 mm for the T group and 1.5 +/- 0.6 mm for the C group). The C group showed more new cementum than the T group (2.3 +/- 0.3 mm versus 2.2 +/- 0.3 mm) as well as a longer epithelium (1.0 +/- 0.3 mm versus 0.9 +/- 0.2 mm). The T group presented more new connective tissue attachment (3.1 +/- 0.2 mm) than the C group (2.9 +/- 0.6 mm). However, no statistically significant differences were detected between the two groups.Conclusions: the amount of new attachment was similar in both groups. Root conditioning with 10% tetracycline solution did not produce any additional new attachment in comparison to the controls.

Immobilization of humic acid in nanostructured layer-by-layer films for sensing applications

Humic acids (HAs), naturally occurring biomacromolecules, were incorporated into nanostructured polymeric films using the layer-by-layer (LbL) technique, in which HA layers were alternated with layers of poly(allylamine hydrochloride) (PAH). Atomic force microscopy (AFM) revealed very smooth films, with mean roughness varying from 0.89 to 1.19 nm for films containing 5 and 15 PAH/HA bilayers, respectively. The films displayed electroactivity, with the presence of only one reduction peak at ca. 0.675 V (vs Ag/AgCl). Such a well-defined electroactivity allowed the films to be used as highly sensitive pesticide sensors, with detection of pentachlorophenol (PCP) in solutions at concentrations as low as 10(-9) mol L(-1).

Molecular engineering strategies to control photo-induced birefringence and surface-relief gratings on layer-by-layer films from an azopolymer

The interactions governing adsorption of layer-by-layer (LBL) films from an azopolymer, PS-119(R), and poly(allylamine hydrochloride) (PAH) have been controlled by modifying the pH of the solutions used to form the films. The thickness per bilayer was varied by one order of magnitude, from 10 to 240 A, as the pH changed from 4 to 10. Thick layers were formed at higher pHs because in this case the PAH is only partially charged adopting a loopy conformation. This change in molecular conformation caused the kinetics of isomerization and mass transport to be much slower than at lower pHs. The writing time defined as the time to achieve 50% of maximum birefringence dropped from 110 to 18 min for films prepared from solutions with pH 10 and pH 4, respectively. This decrease is probably due to the higher free volume for isomerization in the films prepared at lower pHs, in which PAH molecules are less coiled than at higher pHs. For the same reason, the rate of inscription of surface-relief gratings with an interference pattern of p-polarized light was also much slower at higher pHs. (C) 2003 Elsevier B.V. All rights reserved.

Exploiting Cascade Reactions in Bienzyme Layer-by-Layer Films

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Similar anxiolytic-like effects following intra-amygdala infusions of benzodiazepine receptor agonist and antagonist: Evidence for the release of an endogenous benzodiazepine inverse agonist in mice exposed to elevated plus-maze test

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Thermal decomposition of some diuretic agents

Thermogravimetry-derivative thermogravimetry and differential scanning calorimetry were used to study the thermal behaviour of furosemide, hydrochlorothiazide, spironolactone, and amiloride hydrochloride. The results revealed the extents of their thermal stability and also permitted interpretations concerning their thermal decompositions. © 1996 Akadémiai Kiadó.

Characterization of the antinociceptive and sedative effect of amitraz in horses

Amitraz, an acaricide used to control ectoparasites in animals has a complex pharmacological activity, including α2-adrenergic agonist action. The purpose of this research was to investigate the possible antinociceptive and/or sedative effect of amitraz in horses. The sedative effect of the intravenous (i.v.) injection of dimethylformamide (DMF, 5 mL, control) or amitraz (0.05, 0.10, 0.15 mg/kg), was investigated on the head ptosis test. The participation of α2-adrenergic receptors in the sedative effect provoked by amitraz was studied by dosing yohimbine (0.12 mg/kg, i.v.). To measure the antinociception, xylazine hydrochloride (1 mg/kg, i.v., positive control) and the same doses of amitraz and DMF were used. A focused radiant light/heat directed onto the fetlock and withers of a horse were used as a noxious stimulus to measure the hoof withdrawal reflex latency (HWRL) and the skin twitch reflex latency (STRL). The three doses of amitraz used (0.05, 0.10 and 0.15 mg/kg) provoked a dose-dependent relaxation of the cervical muscles. The experiments with amitraz and xylazine on the HWRL showed that after i.v. administration of all doses of amitraz there was a significant increase of HWRL up to 150 min after the injections. Additionally, there was a significant difference between control (DMF) and positive control (xylazine) values up to 30 min after drug injection. On the other hand, the experiments on the STRL show that after administration of amitraz at the dose of 0.15 mg/kg, a significant increase in STRL was observed when compared with the control group. This effect lasted up to 120 min after injection. However, no significant antinociceptive effect was observed with the 0.05 and 0.10 mg/kg doses of amitraz or at the 1.0 mg/kg dose of xylazine.

Morfina nasal: Avaliacao da analgesia e dos efeitos colaterais no tratamento da dor pos-operatoria em criancas

Background and Objectives - Postoperative pain is one of the major discomforts but often under treated, especially in the pediatric patient. The aim of this study was to evaluate nasal morphine postoperative analgesia as an alternative drug administration route and show its applicability, effectiveness, tolerability and side effects. Methods - Participated in this study 20 patients aged 3 to 13 years, physical status ASA I and II sequentially submitted the different small and medium-size surgeries. Analgesia was obtained with nasal morphine hydrochloride in aqueous solution in variable concentrations of 2%, 1%, 0.5%, 0.25% and 0.125%. The dose for each instillation has been 0.1 mg.kg -1 at three-hour intervals for 36 postoperative hours. Quality of analgesia in pre-verbal age patients was evaluated by a pain intensity scale based on facial expression and crying, sleep, motor activity, sociability and food ingestion was used. Standardized evaluations were performed at 3-hour intervals. A four-grade scale was used to evaluate tolerability, where: 1) Good; 2) Regular; 3) Bad; 4) Very bad. Result - Postoperative analgesia results have proven to be good and safe, especially from the third evaluation on (6 hours). Drug tolerability has been good, although side effects were observed, especially nausea and vomiting. Conclusions - Patients and relatives accepted the method very well. The nasal route was considered an adequate way for opioid administration although more studies are needed to accept it as a routine for postoperative morphine analgesia.

Mechanisms of surface-relief gratings formation in layer-by-layer films from azodyes

Surface-relief gratings are photoinscribed on ionically adsorbed layer-by-layer (LBL) films of an azodye, Brilliant Yellow (BY), which was layered alternately with a polyelectrolyte. Photoinscription is performed by impinging an interference pattern of p- or s-polarized laser light with moderate intensity onto the LBL film, which is unlikely to cause thermal effects. Large-scale mass transport occurs due to the force associated with the field gradient of the light pattern. The ionic interactions between adjacent layers appear to provide the means for the chromophores to drag the polymer chains upon photoizomerization. LBL films were produced from two different polyelectrolytes and under two distinct pH values leading to markedly different film properties especially concerning photodegradation. Exposure to the laser light, for instance, leads to higher photodegradation in the poly(dimethyl diallylammonium chloride)/BY system, in comparison to the poly(allylamine hydrochloride)/BY films. Mass transport in the latter case is predominantly light-driven, which is consistent with the higher amplitude of modulation for p-polarized light (70 nm) compared to that caused by s-polarized light (18 nm). © 2003 Elsevier Ltd. All rights reserved.

Competition between humic substances and α-amino acids by metal species

The aim of the present work was to carry out experimental comparison between humic substances (HS) and representative α-amino acids (methionine, methionine sulfoxide and cysteine hydrochloride) in relation to the complexation of biologically active trace elements (Al, Cu, Pb, Mn, Zn, Cd and Ni). A mobile time-controlled tangential-flow UF technique was applied to differentiate between HS-metal and α-aminoacids-metal complexes. Metal determinations were conventionally carried out using a ICP-OES. The results showed that HS may be considered as a selective complexing agents with higher metal bonding capability in relation to Al, Cu and Pb, the fact that may be clinically important.

An alternative LC-UV procedure for the determination of prochloraz residues in fruits

An alternative method using liquid chromatography with UV detection for the determination of prochloraz as 2,4,6-trichlorophenol in mango, papaya and orange is described. Ethyl acetate, acetone and dichloromethane were tested for extraction of prochloraz from the fruits. After extraction the residue of prochloraz was derivatized with pyridine hydrochloride. The analysis was carried out using liquid chromatography with UV detection and gas chromatography with electron-capture detection. Average recoveries of prochloraz from spiked fruits (0.1 and 0.2 mg kg-1) ranged from 80% to 94% with relative standard deviations between 5.6% and 12.6% (n=8). Detection and quantification limits were 0.05 and 0.1 mg kg-1, respectively. The LC-UV method was applied to mango and papaya samples submitted to dip treatment with a prochloraz formulation under laboratory conditions. In addition, fruit samples obtained from local markets were analysed. ©2005 Sociedade Brasileira de Química.