Ocular biocompatibility of novel Cyclosporin A formulations based on methoxy poly(ethylene glycol)-hexylsubstituted poly(lactide) micelle carriers.

Topical ocular drug delivery has always been a challenge for pharmaceutical technology scientists. In the last two decades, many nano-systems have been studied to find ways to overcome the typical problems of topical ocular therapy, such as difficult corneal penetration and poor drug availability. In this study, methoxy poly(ethylene glycol)-hexylsubstituted poly(lactides) (MPEG-hexPLA) micelle formulations, which are promising nanocarriers for poorly water soluble drugs, were investigated for the delivery of Cyclosporin A (CsA) to the eye. As a new possible pharmaceutical excipient, the ocular compatibility of MPEG-hexPLA micelle formulations was evaluated. An in vitro biocompatibility assessment on human corneal epithelial cells was carried out using different tests. Cytotoxicity was studied by using the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] (MTT), and clonogenic tests and revealed that the CsA formulations and copolymer solutions were not toxic. After incubation with MPEG-hexPLA micelle formulations, the activation of caspase-dependent and -independent apoptosis as well as autophagy was evaluated using immunohistochemistry by analyzing the localization of four antibodies: (1) anti-caspase 3; (2) anti-apoptotic inducing factor (AIF); (3) anti-IL-Dnase II and (4) anti-microtubule-associated protein 1 light chain 3 (LC3). No apoptosis was induced when the cells were treated with the micelle solutions that were either unloaded or loaded with CsA. The ocular tolerance was assessed in vivo on rabbit eyes by Confocal Laser Scanning Ophthalmoscopy (CLSO), and very good tolerability was seen. The observed corneal surface was comparable to a control surface that was treated with a 0.9% NaCl solution. In conclusion, these results demonstrate that MPEG-hexPLA micelles are promising drug carriers for ocular diseases involving the activation of cytokines, such as dry eye syndrome and autoimmune uveitis, or for the prevention of corneal graft rejection.

A comparison of in vivo 13C MR brain glycogen quantification at 9.4 and 14.1 T.

The high molecular weight and low concentration of brain glycogen render its noninvasive quantification challenging. Therefore, the precision increase of the quantification by localized (13) C MR at 9.4 to 14.1 T was investigated. Signal-to-noise ratio increased by 66%, slightly offset by a T(1) increase of 332 ± 15 to 521 ± 34 ms. Isotopic enrichment after long-term (13) C administration was comparable (≈ 40%) as was the nominal linewidth of glycogen C1 (≈ 50 Hz). Among the factors that contributed to the 66% observed increase in signal-to-noise ratio, the T(1) relaxation time impacted the effective signal-to-noise ratio by only 10% at a repetition time = 1 s. The signal-to-noise ratio increase together with the larger spectral dispersion at 14.1 T resulted in a better defined baseline, which allowed for more accurate fitting. Quantified glycogen concentrations were 5.8 ± 0.9 mM at 9.4 T and 6.0 ± 0.4 mM at 14.1 T; the decreased standard deviation demonstrates the compounded effect of increased magnetization and improved baseline on the precision of glycogen quantification.

CRESCIMENTO E RELAÇÕES HÍDRICAS EM PLANTAS DE GOIABEIRA SUBMETIDAS A ESTRESSE SALINO COM NaCl

Tendo como objetivo determinar os efeitos do estresse salino sobre o crescimento e as relações hídricas de plantas de goiabeira (Psidium guajava L), conduziu-se um experimento em casa de vegetação, em solução nutritiva, na Universidade Federal do Ceará, no período de setembro a dezembro de 1997. Os tratamentos consistiram de sete níveis de NaCl (0; 25; 50; 75; 100; 125 e 150 mmol L-1). Avaliaram-se, aos 50 dias após o estresse salino (DAE): altura da planta, número de folhas e área foliar unitária e por planta. A taxa de crescimento relativo (TCR) foi determinada entre 30 e 50 DAE. Aos 20; 30; 40 e 50 DAE, foram determinadas condutâncias estomáticas, transpiração, potencial hídrico e teor relativo de água (TRA). Dentre as variáveis estudadas, a área foliar por planta foi a mais afetada, com uma redução de 92% quando submetida ao nível mais elevado de salinidade (150 mmol L-1). A condutância estomática, a transpiração, o potencial hídrico foliar (psiwf) e o teor relativo de água (TRA) decresceram com o aumento dos níveis e do tempo de exposição ao NaCl. As plantas jovens de goiabeira são sensíveis à salinidade, com uma salinidade limiar de 1,2 dSm-1. A taxa de sobrevivência foi reduzida em 75% quando as plantas foram submetidas a 150 mmol L-1 até 50 dias após o estresse.

Intravoxel incoherent motion diffusion-weighted imaging in the liver: comparison of mono-, bi- and tri-exponential modelling at 3.0-T.

PURPOSE: To determine whether a mono-, bi- or tri-exponential model best fits the intravoxel incoherent motion (IVIM) diffusion-weighted imaging (DWI) signal of normal livers. MATERIALS AND METHODS: The pilot and validation studies were conducted in 38 and 36 patients with normal livers, respectively. The DWI sequence was performed using single-shot echoplanar imaging with 11 (pilot study) and 16 (validation study) b values. In each study, data from all patients were used to model the IVIM signal of normal liver. Diffusion coefficients (Di ± standard deviations) and their fractions (fi ± standard deviations) were determined from each model. The models were compared using the extra sum-of-squares test and information criteria. RESULTS: The tri-exponential model provided a better fit than both the bi- and mono-exponential models. The tri-exponential IVIM model determined three diffusion compartments: a slow (D1 = 1.35 ± 0.03 × 10(-3) mm(2)/s; f1 = 72.7 ± 0.9 %), a fast (D2 = 26.50 ± 2.49 × 10(-3) mm(2)/s; f2 = 13.7 ± 0.6 %) and a very fast (D3 = 404.00 ± 43.7 × 10(-3) mm(2)/s; f3 = 13.5 ± 0.8 %) diffusion compartment [results from the validation study]. The very fast compartment contributed to the IVIM signal only for b values ≤15 s/mm(2) CONCLUSION: The tri-exponential model provided the best fit for IVIM signal decay in the liver over the 0-800 s/mm(2) range. In IVIM analysis of normal liver, a third very fast (pseudo)diffusion component might be relevant. KEY POINTS: ? For normal liver, tri-exponential IVIM model might be superior to bi-exponential ? A very fast compartment (D = 404.00 ± 43.7 × 10 (-3)  mm (2) /s; f = 13.5 ± 0.8 %) is determined from the tri-exponential model ? The compartment contributes to the IVIM signal only for b ≤ 15 s/mm (2.)

Inoculação e sobrevivência de Xanthomonas vesicatoria em sementes de tomateiro

Para avaliar o efeito de métodos de inocuação de Xanthomonas vesicatoria (Doidge) (9) em sementes de tomate sobre a qualidade da semente e transmissão da fitobactéria compararam-se os tratamentos: 1) inoculação a vácuo com suspensão de células de X. vesicatoria em STP (0,005M, pH 7,4 e NaCl 0,85%); 2) imersão por 24 horas em suspensão de células de X. vesicatoria em STP; 3) vácuo em solução STP; 4) imersão por 24 horas em STP; 5) imersão por 5 min. em álcool etílico e 6) semente original. As avaliações foram realizadas por testes de germinação e isolamentos em meio Nutriente Agar Modificado (NAM) aos 1, 15, 30 e 45 dias após aplicação dos tratamentos. Em seguida, avaliou-se o efeito da umidade (4% e 8%) e o armazenamento das sementes inoculadas e variações de umidade antes da realização dos testes sobre a sobrevivência e recuperação de X. vesicatoria. Os métodos de inoculação testados podem ser utilizados em trabalhos de rotina, porém, as sementes devem ser utilizadas até 30 dias após a inoculação a partir de quando ocorre uma redução acentuada na taxa de recuperação da fitobactéria. A umidade das sementes interfere na sobrevivência e na transmissão de X. vesicatoria pelas sementes de tomate.

Viabilidade de armazenamento de sementes de soja inoculadas com Sclerotinia sclerotiorum em meio com restrição hídrica

O trabalho objetivou avaliar o efeito da restrição hídrica, em meio de cultura agarizado, sobre a germinação de sementes de soja inoculadas com Sclerotinia sclerotiorum nos potenciais osmóticos de 0, -0,3, -0,6 e -0,9 MPa promovido pelo uso dos solutos manitol, MgCl e NaCl, em diferentes tempos de exposição das sementes ao patógeno (6, 12, 18, 24 e 30 horas) e a possibilidade de armazenamento das sementes inoculadas. Avaliaram-se o crescimento micelial de S. sclerotiorum nos meios modificados osmoticamente, a germinação das sementes submetidas à inoculação e, após o armazenamento das sementes inoculadas, a emergência em solo e transmissão do patógeno. Pelos resultados obtidos a adição de manitol ao meio de cultura BDA, nos potenciais osmóticos -0,3, -0,6 e -0,9 MPa, e de NaCl, -0,3 e -0,6 MPa, não restringe o crescimento micelial de S. sclerotiorum; os solutos utilizados até o potencial osmótico -0,9 MPa não interferiram na germinação de sementes de soja. Níveis de infecção satisfatórios foram obtidos com 24 horas de incubação em meio de cultura com restrição hídrica. Tanto a viabilidade das sementes como a do patógeno são mantidas após o armazenamento das sementes inoculadas em meio com restrição hídrica.

Citologia vaginal a fresco na gravidez: correlação com a citologia corada pela técnica de Papanicolaou

OBJETIVO: analisar o conteúdo vaginal utilizando o exame citológico a fresco na primeira consulta pré-natal em mulheres com ou sem queixas genitais e correlacionar os resultados com os encontrados na citologia corada pela técnica de Papanicolaou. A microscopia direta durante a gravidez deve ser valorizada e reconhecida como método propedêutico capaz de diagnosticar, de forma imediata, 90% dos casos de vaginose bacteriana, candidose e tricomonose. MÉTODOS: estudo prospectivo em 216 gestantes, selecionadas em ambulatório de pré-natal no período de 30 de outubro de 2001 a 12 de novembro de 2002. Foram colhidas duas amostras do conteúdo existente no fundo de saco vaginal posterior e depositadas em lâminas de vidro para microscopia. Sobre a primeira e a segunda amostra eram colocadas uma gota de NaCl a 0,9% e uma de KOH a 10%, respectivamente. Adicionalmente, em todas as grávidas determinou-se o pH vaginal e realizaram-se os testes de produção das aminas com odores de pescado. O material era examinado ao microscópio em aumentos de 100 vezes, 400 vezes e excepcionalmente 1000 vezes. Foram realizados esfregaços cervicovaginais para citologia corada pelo método de Papanicolaou. A correlação entre os resultados dos métodos citológicos empregados foi realizada pelo cálculo do coeficiente kappa, que avalia a concordância para variáveis qualitativas. RESULTADOS: o encontro nos esfregaços a fresco de flora bacteriana normal foi de 51,8, representando o aspecto citológico mais observado e sem correspondência com os 3,7% apurados na microscopia corada. No exame citológico direto foram observados 30,9% de vaginose bacteriana e 7,9% de candidose. Todavia, no Papanicolaou não foi encontrada tal equivalência, sendo as porcentagens de 0,7 e 24,3%, respectivamente. A ausência de correlação no diagnóstico de colpite bacteriana inespecífica na microscopia direta (17,5%) e corada (51,3%) talvez deva-se ao subdiagnóstico de vaginose neste último método propedêutico. Os diagnósticos de tricomoníase observados em ambos os métodos citológicos (3,7 e 2,7%) traduzem a baixa prevalência destes parasitas na gestação. O cálculo do índice kappa para avaliação da concordância entre os dois procedimentos citológicos nos diversos achados microbiológicos demonstrou baixa correlação nos diagnósticos da vaginose bacteriana e colpites bacterianas inespecíficas, bem como na identificação da flora vaginal normal. CONCLUSÕES: embora a citologia corada apresente melhor acurácia no diagnóstico de fungos não formadores de micélios, a citologia direta a fresco demonstrou ser melhor avaliador dos elementos não epiteliais dos esfregaços vaginais. Entretanto, a microscopia corada pela técnica de Papanicolaou, por permitir melhor apreciação das células epiteliais cervicovaginais, representa o mais importante instrumento revelador das agressões e reações nucleocitoplasmáticas.

Deposição e distribuição de calda de pulverização em plantas de feijoeiro (Phaseolus vulgaris) e capim-braquiária (Brachiaria decumbens)

Com o objetivo de avaliar a deposição e distribuição de solução traçante em plantas de feijoeiro e capim-braquiária, foi conduzido experimento no NuPAM-FCA/ UNESP, campus de Botucatu-SP, utilizando um simulador de pulverização em ambiente controlado. Os tratamentos utilizados foram: bico jato plano XR Teejet (XR110.02VS) + surfactante Aterbane BR (0,5% v/v); bico jato cônico Conejet (TXVK-4) + surfactante Aterbane BR (0,5% v/v); bico jato plano XR Teejet (XR110.02VS); e bico jato cônico Conejet (TXVK-4). Em todos os tratamentos foi aplicada uma solução de NaCl (0,5% p/v) + corante Poliglow laranja (0,5% p/v) + mancozeb (0,5% p/v). As unidades experimentais constituíram-se de vasos com duas plantas-alvo de capim-braquiária posicionadas sob uma planta-alvo de feijoeiro. A visualização da distribuição das gotas nas folhas de capim-braquiária foi efetuada com auxílio de luz negra, e o depósito da calda na superfície das plantas foi quantificado através da condutividade elétrica da solução aplicada e coletada por meio de lavagem de ambos os alvos. Os bicos de pulverização, jato plano (XR Teejet) e cônico (Conejet), não apresentaram diferença no depósito nos folíolos totais de feijoeiro quando submetidos a mesma condição de calda de pulverização. No entanto, o bico jato plano XR Teejet (XR110.02VS) e o bico jato cônico Conejet (TXVK-4) proporcionaram aumento de 67,1 e 61,5% na deposição da calda em relação a área foliar e 106,4 e 66,9% para matéria seca, respectivamente, em relação ao bico jato plano XR Teejet (XR110.02VS) + surfactante Aterbane BR (0,5% v/v) e ao bico jato cônico Conejet (TXVK-4) + surfactante Aterbane BR (0,5% v/v). Para o capim-braquiária, o bico cônico Conejet proporcionou deposição superior e distribuição mais uniforme em relação ao jato plano XR Teejet.

Fatores ambientais sobre a germinação de Emilia sonchifolia

O conhecimento de informações básicas sobre a biologia de plantas daninhas pode contribuir significativamente para o desenvolvimento de estratégias para seu manejo. Emilia sonchifolia é uma espécie de ampla disseminação em todo o território brasileiro e apresenta moderada agressividade, sendo encontrada em áreas agrícolas, terrenos baldios e áreas urbanas. O presente trabalho teve como objetivo avaliar a germinação das sementes de E. sonchifolia em diferentes condições ambientais. Foram realizados quatro experimentos: combinação de quatro temperaturas (20, 25, 30 e 35 ºC) em duas condições de luz (ausência e presença); sete níveis de restrição hídrica proporcionada por PEG6000 (0,0; -0,1; -0,2; -0,4; -0,6; -0,8; e -1,0 MPa); seis níveis de estresse salino proporcionado pelo NaCl (0,0; -0,2; -0,4; -0,6; -0,8; e -1,0 MPa) e seis níveis de pH (3,0; 5,0; 6,0; 7,0; 9,0; e 11,0) em solução tamponada. A germinação das sementes de E. sonchifolia foi maximizada na presença de luz, em temperaturas constantes de 25 e 30 ºC. Houve redução da germinação em ambiente com restrição hídrica a partir de -0,1 MPa, e para estresse salino, a partir de 0,2 MPa. E. sonchifolia é sensível à variação de pH, ocorrendo maior germinação das sementes apenas em pH 6,0.

Effect of bacterial lipopolysaccharide on gastric emptying of liquids in rats

The objectives of the present investigation were 1) to study the effect of bacterial lipopolysaccharide (LPS) on rat gastric emptying (GE) and 2) to investigate a possible involvement of the vagus nerve in the gastric action of LPS. Endotoxin from E. coli (strain 055:B5) was administered sc, ip or iv to male Wistar rats (220-280 g body weight) at a maximum dose of 50 µg/kg animal weight. Control animals received an equivalent volume of sterile saline solution. At a given time period after LPS administration, GE was evaluated by measuring gastric retention 10 min after the orogastric infusion of a test meal (2 ml/100 g animal weight), which consisted of 0.9% NaCl plus the marker phenol red (6 mg/dl). One group of animals was subjected to bilateral subdiaphragmatic vagotomy or sham operation 15 days before the test. A significant delay in GE of the test meal was observed 5 h after iv administration of the endotoxin at the dose of 50 µg/kg animal weight. The LPS-induced delay of GE was detected as early as 30 min and up to 8 h after endotoxin administration. The use of different doses of LPS ranging from 5 to 50 µg/kg animal weight showed that the alteration of GE was dose dependent. In addition, vagotomized animals receiving LPS displayed a GE that was not significantly different from that of the sham control group. However, a participation of the vagus nerve in LPS-induced delay in GE could not be clearly demonstrated by these experiments since vagotomy itself induced changes in this gastric parameter. The present study provides a suitable model for identifying the mechanisms underlying the effects of LPS on gastric emptying

Release of plasma atrial natriuretic peptide after volume expansion is not related to pituitary-adrenal axis diurnal variation in normal subjects

The existence of a circadian rhythm of atrial natriuretic peptide (ANP) in humans is controversial. We studied the plasma ANP response to isotonic blood volume expansion in the morning and in the afternoon and its relationship with adrenocorticotropic hormone (ACTH)-cortisol diurnal variation in seven normal subjects. Basal plasma ANP level was similar in the morning (19.6 ± 2.4 pg/ml) and in the afternoon (21.8 ± 4.8 pg/ml). The ANP peak obtained with saline infusion (0.9% NaCl, 12 ml/kg) in the morning (49.4 ± 8 pg/ml) did not differ from that obtained in the afternoon (60.3 ± 10.1 pg/ml). There was no correlation between the individual mean cortisol and ACTH levels and the ANP peak obtained with saline infusion. These data indicate no diurnal variation in plasma ANP secretion induced by blood volume expansion and no relationship between plasma ANP peak and ACTH-cortisol diurnal variation

Acute extracellular fluid volume changes increase ileocolonic resistance to saline flow in anesthetized dogs

We determined the effect of acute extracellular fluid volume changes on saline flow through 4 gut segments (ileocolonic, ileal, ileocolonic sphincter and proximal colon), perfused at constant pressure in anesthetized dogs. Two different experimental protocols were used: hypervolemia (iv saline infusion, 0.9% NaCl, 20 ml/min, volume up to 5% body weight) and controlled hemorrhage (up to a 50% drop in mean arterial pressure). Mean ileocolonic flow (N = 6) was gradually and significantly decreased during the expansion (17.1%, P<0.05) and expanded (44.9%, P<0.05) periods while mean ileal flow (N = 7) was significantly decreased only during the expanded period (38%, P<0.05). Mean colonic flow (N = 7) was decreased during expansion (12%, P<0.05) but returned to control levels during the expanded period. Mean ileocolonic sphincter flow (N = 6) was not significantly modified. Mean ileocolonic flow (N = 10) was also decreased after hemorrhage (retracted period) by 17% (P<0.05), but saline flow was not modified in the other separate circuits (N = 6, 5 and 4 for ileal, ileocolonic sphincter and colonic groups, respectively). The expansion effect was blocked by atropine (0.5 mg/kg, iv) both on the ileocolonic (N = 6) and ileal (N = 5) circuits. Acute extracellular fluid volume retraction and expansion increased the lower gastrointestinal resistances to saline flow. These effects, which could physiologically decrease the liquid volume being supplied to the colon, are possible mechanisms activated to acutely balance liquid volume deficit and excess.

Faster gastric emptying of a liquid meal in rats after hypothalamic dorsomedial nucleus lesion

The effects of dorsomedial hypothalamic (DMH) nucleus lesion on body weight, plasma glucose levels, and the gastric emptying of a liquid meal were investigated in male Wistar rats (170-250 g). DMH lesions were produced stereotaxically by delivering a 2.0-mA current for 20 s through nichrome electrodes (0.3-mm tip exposure). In a second set of experiments, the DMH and the ventromedial hypothalamic (VMH) nucleus were lesioned with a 1.0-mA current for 10 s (0.1-mm tip exposure). The medial hypothalamus (MH) was also lesioned separately using a nichrome electrode (0.3-mm tip exposure) with a 2.0-mA current for 20 s. Gastric emptying was measured following the orogastric infusion of a liquid test meal consisting of physiological saline (0.9% NaCl, w/v) plus phenol red dye (6 mg/dl) as a marker. Plasma glucose levels were determined after an 18-h fast before the lesion and on the 7th and 15th postoperative day. Body weight was determined before lesioning and before sacrificing the rats. The DMH-lesioned rats showed a significantly faster (P<0.05) gastric emptying (24.7% gastric retention, N = 11) than control (33.0% gastric retention, N = 8) and sham-lesioned (33.5% gastric retention, N = 12) rats, with a transient hypoglycemia on the 7th postoperative day which returned to normal by the 15th postoperative day. In all cases, weight gain was slower among lesioned rats. Additional experiments using a smaller current to induce lesions confirmed that DMH-lesioned rats had a faster gastric emptying (25.1% gastric retention, N = 7) than control (33.4% gastric retention, N = 17) and VMH-lesioned (34.6% gastric retention, N = 7) rats. MH lesions resulted in an even slower gastric emptying (43.7% gastric retention, N = 7) than in the latter two groups. We conclude that although DMH lesions reduce weight gain, they do not produce consistent changes in plasma glucose levels. These lesions also promote faster gastric emptying of an inert liquid meal, thus suggesting a role for the DMH in the regulation of gastric motility

Effect of selective angiotensin antagonists on the antidiuresis produced by angiotensin-(1-7) in water-loaded rats

In the present study we evaluated the nature of angiotensin receptors involved in the antidiuretic effect of angiotensin-(1-7) (Ang-(1-7)) in water-loaded rats. Water diuresis was induced in male Wistar rats weighing 280 to 320 g by water load (5 ml/100 g body weight by gavage). Immediately after water load the rats were treated subcutaneously with (doses are per 100 g body weight): 1) vehicle (0.05 ml 0.9% NaCl); 2) graded doses of 20, 40 or 80 pmol Ang-(1-7); 3) 200 nmol Losartan; 4) 200 nmol Losartan combined with 40 pmol Ang-(1-7); 5) 1.1 or 4.4 nmol A-779; 6) 1.1 nmol A-779 combined with graded doses of 20, 40 or 80 pmol Ang-(1-7); 7) 4.4 nmol A-779 combined with graded doses of 20, 40 or 80 pmol Ang-(1-7); 8) 95 nmol CGP 42112A, or 9) 95 nmol CGP 42112A combined with 40 pmol Ang-(1-7). The antidiuretic effect of Ang-(1-7) was associated with an increase in urinary Na+ concentration, an increase in urinary osmolality and a reduction in creatinine clearance (CCr: 0.65 ± 0.04 ml/min vs 1.45 ± 0.18 ml/min in vehicle-treated rats, P<0.05). A-779 and Losartan completely blocked the effect of Ang-(1-7) on water diuresis (2.93 ± 0.34 ml/60 min and 3.39 ± 0.58 ml/60 min, respectively). CGP 42112A, at the dose used, did not modify the antidiuretic effect of Ang-(1-7). The blockade produced by Losartan was associated with an increase in CCr and with an increase in sodium and water excretion as compared with Ang-(1-7)-treated rats. When Ang-(1-7) was combined with A-779 there was an increase in CCr and natriuresis and a reduction in urine osmolality compared with rats treated with Ang-(1-7) alone. The observation that both A-779, which does not bind to AT1 receptors, and Losartan blocked the effect of Ang-(1-7) suggests that the kidney effects of Ang-(1-7) are mediated by a non-AT1 angiotensin receptor that is recognized by Losartan.

Kallikrein-like amidase activity in renal ischemia and reperfusion

We assessed a kallikrein-like amidase activity probably related to the kallikrein-kinin system, as well as the participation of leukocyte infiltration in renal ischemia and reperfusion. Male C57BL/KSJmdb mice were subjected to 20 or 60 min of ischemia and to different periods of reperfusion. A control group consisted of sham-operated mice, under similar conditions, except for ischemia induction. Kallikrein-like amidase activity, Evans blue extravasation and myeloperoxidase activity were measured in kidney homogenates, previously perfused with 0.9% NaCl. Plasma creatinine concentration increased only in the 60-min ischemic group. After 20 min of ischemia and 1 or 24 h of reperfusion, no change in kallikrein-like amidase activity or Evans blue extravasation was observed. In the mice subjected to 20 min of ischemia, edema was evident at 1 h of reperfusion, but kidney water content returned to basal levels after 24 h of reperfusion. In the 60-min ischemic group, kallikrein-like amidase activity and Evans blue extravasation showed a similar significant increase along reperfusion time. Kallikrein-like amidase activity increased from 4 nmol PNA mg protein-1 min-1 in the basal condition to 15 nmol PNA mg protein-1 min-1 at 10 h of reperfusion. For dye extravasation the concentration measured was near 200 µg of Evans blue/g dry tissue in the basal condition and 1750 µg of Evans blue/g dry tissue at 10 h of reperfusion. No variation could be detected in the control group. A significant increase from 5 to 40 units of DAbs 655 nm g wet tissue-1 min-1 in the activity of the enzyme myeloperoxidase was observed in the 60-min ischemic group, when it was evaluated after 24 h of reperfusion. Histological analysis of the kidneys showed migration of polymorphonuclear leukocytes from the vascular bed to the interstitial tissue in the 60-min ischemic group after 24 h of reperfusion. We conclude that the duration of ischemia is critical for the development of damage during reperfusion and that the increase in renal cortex kallikrein-like amidase activity probably released from both the kidney and leukocytes may be responsible, at least in part, for the observed effects, probably through direct induction of increased vascular permeability.