ITS2 as a molecular marker for the identification of Diatraea saccharalis and D. flavipennella and possible infection with Cotesia spp.

In Brazil, the species Diatraea flavipennella and D. saccharalis play an important role in the sugar and alcohol agribusiness by causing many damages in sugarcane fields. The egg, larva, pupa, and adult stages are very morphologically similar between these species, and the identification can be confused. The internal transcribed spacer 2 (ITS 2) from ribosomal DNA has important features as evolutionary divergence. It is a good marker for species identification, participates in the rDNA processing, and has been applied in phylogenetic and population studies. This study aimed to make available a molecular marker to assist on the identification method of pests' species of Diatraea and to identify possible traces of Cotesia in the resistant host. The DNA was extracted from the egg, larva, and adult samples. PCR amplicons were purified and sequenced. The sequences were analyzed in MEGA 5.01. The ITS 2 length was 410 bp in D. flavipennella and 448 bp in D. saccharalis. The GC content was similar between the species. Three microsatellite loci were present in D. saccharalis and absent in D. flavipennella, contributing to differences in ITS 2 length in the species. An additional 367-bp band was attributed to Cotesia spp. The differences among ITS 2 from D. flavipennella, D. saccharalis, and Cotesia sp were sufficient to identify them on electrophoresis gel and sequencing. The presence of Cotesia sp traits in adult D. flavipennella showed possible host refractoriness, but further studies are necessary.

BTCI enhances guanylin-induced natriuresis and promotes renal glomerular and tubular effects.

Guanylin and uroguanylin are small cysteine-rich peptides involved in the regulation of fluid and electrolyte homeostasis through binding and activation of guanylyl cyclases signaling molecules expressed in intestine and kidney. Guanylin is less potent than uroguanylin as a natriuretic agent and is degraded in vitro by chymotrypsin due to unique structural features in the bioactive moiety of the peptide. Thus, the aim of this study was to verify whether or not guanylin is degraded by chymotrypsin-like proteases present in the kidney brush-border membranes. The isolated perfused rat kidney assay was used in this regard. Guanylin (0.2 microM) induced no changes in kidney function. However, when pretreated by the black-eyed pea trypsin and chymotrypsin inhibitor (BTCI - 1.0 microM; guanylin - 0.2 microM) it promoted increases in urine flow (DeltaUF of 0.25 +/- 0.09 mL.g(-1)/min, P < 0.05) and Na+ excretion (% Delta ENa+ of 18.20 +/- 2.17, P < 0.05). BTCI (1.0 microM) also increased %ENa+ (from 22.8 +/- 1.30 to 34.4 +/- 3.48, P < 0.05, 90 minutes). Furthermore, BTCI (3.0 microM) induced increases in glomerular filtration rate (GFR; from 0.96 +/- 0.02 to 1.28 0.02 mL.g(-1)/min, P < 0.05, 60 minutes). The present paper strongly suggests that chymotrypsin-like proteases play a role in renal metabolism of guanylin and describes for the first time renal effects induced by a member of the Bowman-Birk family of protease inhibitors.

BTCI enhances guanylin-induced natriuresis and promotes renal glomerular and tubular effects

Guanylin and uroguanylin are small cysteine-rich peptides involved in the regulation of fluid and electrolyte homeostasis through binding and activation of guanylyl cyclases signaling molecules expressed in intestine and kidney. Guanylin is less potent than uroguanylin as a natriuretic agent and is degraded in vitro by chymotrypsin due to unique structural features in the bioactive moiety of the peptide. Thus, the aim of this study was to verify whether or not guanylin is degraded by chymotrypsin-like proteases present in the kidney brush-border membranes. The isolated perfused rat kidney assay was used in this regard. Guanylin (0.2 µM) induced no changes in kidney function. However, when pretreated by the black-eyed pea trypsin and chymotrypsin inhibitor (BTCI - 1.0 µM; guanylin - 0.2 µM) it promoted increases in urine flow (deltaUF of 0.25 ± 0.09 mL.g-1/min, P < 0.05) and Na+ excretion ( percent delta ENa+ of 18.20 ± 2.17, P < 0.05). BTCI (1.0 µM) also increased percentENa+ (from 22.8 ± 1.30 to 34.4 ± 3.48, P < 0.05, 90 minutes). Furthermore, BTCI (3.0 µM) induced increases in glomerular filtration rate (GFR; from 0.96 ± 0.02 to 1.28 0.02 mL.g-1/min, P < 0.05, 60 minutes). The present paper strongly suggests that chymotrypsin-like proteases play a role in renal metabolism of guanylin and describes for the first time renal effects induced by a member of the Bowman-Birk family of protease inhibitors.

Guanilina e uroguanilina são peptídeos pequenos, ricos em cisteína, envolvidos na regulação da homeostase de fluidos e eletrólitos através da ligação e ativação da guanilato ciclase expressa no intestino e nos rins. A guanilina é menos potente do que a uroguanilina como agente natriurético e é degradada in vitro pela quimiotripsina devido a características estruturais únicas no domínio bioativo do peptídeo. Portanto o objetivo deste trabalho foi verificar se a guanilina é degradada por proteases tipo quimiotripsina, presentes na membrana da borda em escova dos rins. Para esta investigação, foi usado o modelo do rim isolado de rato perfundido. A Guanilina (0,2 µM) não induziu mudanças na função renal. Entretanto, quando pré-tratada com inibidor de tripsina e de quimiotripsina de black-eyed pea (BTCI - 1,0 µM; guanilina - 0,2 µM) promoveu um aumento no fluxo urinário (deltaUF de 0,25 ± 0,09 mL.g-1/min, P < 0,05) e na excreção de Na+ ( por centoDENa+ de 18,20 ± 2,17, P < 0,05). BTCI (1,0 µM) também aumenta por centoENa+ (de 22,8 ± 1,30 a 34,4 ± 3,48, P < 0,0590 minutos). Além disto, BTCI (3,0 µM) induziu um aumento da taxa de filtração glomerular (GFR; de 0,96 ± 0,02 para 1,28 ± 0,02 mL.g-1/min, P < 0,05, 60 minutos). O presente trabalho sugere fortemente que proteases semelhantes à quimiotripsina desempenham um papel no metabolismo renal de guanilinas e descreve, pela primeira vez, os efeitos renais induzidos por um membro da família de inibidores de proteases do tipo Bowman-Birk.

Characterization and immunogenicity of meningococcal group C conjugate vaccine prepared using hydrazide-activated tetanus toxoid.

The steps to produce, purify and control an immunogenic Brazilian conjugate vaccine against group C meningococcus (MenCPS-TT) using hydrazide-activated tetanus toxoid were developed. The conjugation methodology reduced the reaction time easily allowing scale-up. One freeze-dried pilot vaccine lot purified by tangential filtration, showed satisfactory quality control results including safety and stability. The pilot vaccine was immunogenic in mice in a dose-dependent fashion generating a 10-20-fold rise in IgG response in mice. The vaccine also induced high bactericidal titers. Vaccine concentrations of 1 and 0.1 microg showed higher avidity indices, suggesting induction of immunologic memory. These results support initiation of Phase I clinical studies with the MenCPS-TT conjugate vaccine.

Aplicaçäo clínica do retalho fáscio-cutâneo da regiäo posterior da coxa em V-Y / Clinical application of the VY posterior thigh fasciocutaneous flap

OBJETIVO: Analisar a aplicabilidade do retalho fáscio-cutâneo em V-Y da regiäo posterior da coxa na reparaçäo de lesöes isoladas ou múltiplas nas regiöes glútea e perineal. MÉTODO: Foram submetidos a tratamento cirúrgico 20 pacientes, portadores de 25 úlceras nas regiöes glútea e perineal, sendo 23 delas úlceras por pressäo, um pós-abscesso perianal e um pós-trauma perineal. Em todos os pacientes a reparaçäo foi realizada com o retalho fáscio-cutâneo da regiäo posterior da coxa. O retalho foi confeccionado com base súpero-lateral, preservando-se os ramos fáscio-cutâneos das artérias glútea inferior, primeira e segunda perfurantes no tratamento de úlceras isquiáticas. Nas associaçöes de úlcera isquiática com úlcera sacral, adicionou-se ao retalho uma extensäo fáscio-cutânea da regiäo glútea para tratamento cirúrgico em tempo único. Foi realizado retalho com base súpero-medial, preservando-se o ramo fáscio-cutâneo da artéria glútea inferior no tratamento das úlceras trocantéricas. Nas associaçöes com úlcera sacral, acrescentou-se ao retalho posterior da coxa uma extensäo fáscio-cutânea da regiäo glútea, que permitiu o fechamento de todas as úlceras em um só tempo cirúrgico. Nas associaçöes de úlceras trocantéricas e isquiáticas, realizou-se o retalho com base superior, preservando-se o ramo fáscio-cutâneo da artéria glútea inferior. RESULTADOS: Näo houve necrose do retalho. As complicaçöes imediatas foram três infecçöes, uma deiscência e um hematoma. Num período de seguimento de 6 meses a 29 meses, houve recidiva de seis úlceras (6/24 = 25,0 por cento) em cinco pacientes (5/19 = 26,32 por cento). Um paciente foi perdido neste seguimento. CONCLUSÄO: Conclui-se que o retalho fáscio-cutâneo posterior da coxa, em V-Y, pode ser utilizado com segurança no tratamento de lesöes isoladas ou múltiplas das regiöes glútea e perineal

[Clinical application of the V-Y posterior thigh fasciocutaneous flap]. / Aplicação clínica do retalho fáscio-cutâneo da região posterior da coxa em V-Y.

PURPOSE: To analyze the clinical applicability of one-stage treatment of single or multiple sores using the V-Y posterior thigh fasciocutaneous flap. METHODS: Twenty patients were treated, having 25 ulcers in the gluteal and perineal regions, being 23 pressure ulcers, 1 perianal abscess and 1 perineal trauma. In all of the patients the repair was carried out with a fasciocutaneous flap from the posterior region of the thigh. The flap was made with a superior and lateral base, preserving the fasciocutaneous branches of the inferior gluteal arteries, first and second perforating, in the treatment of ischial ulcers. In the associations of the same with sacral ulcer, a fasciocutaneous extension from the gluteal region was added to the flap for a one and only surgical procedure. A superior and medial based flap was made, preserving the fasciocutaneous branch of the inferior gluteal artery in the treatment of the trochanteric ulcers. In the associations with sacral ulcer a fasciocutaneous extension from the gluteal region was added to the posterior thigh flap which permitted the closure of all ulcers in only one surgical procedure. In the associations of trochanteric and ischial ulcers a flap with a superior base was made, preserving the fasciocutaneous branch of the inferior gluteal artery. RESULTS: There was no necrosis of the flap. The immediate complications were 3 infections, 1 dehiscences and 1 hematoma. In a follow-up period of 6 months to 29 months, with one patient being lost to follow up, there were 6 recurrences 6/24 (25.0%) in 5 patients 5/19 (26.32%). CONCLUSION: We conclude that the posterior fasciocutaneous thigh flap, in V-Y, can be used with safety in the treatment of isolated or multiple ulcers in the gluteal and perineal regions.

Hepatitis B and hepatitis C prevalence among blood donors and HIV-1 infected patients in Florianópolis--Brazil.

Information is scarce on the prevalence of hepatitis B (HBV) and hepatitis C (HCV) among voluntary blood donors and patients infected with the human immunodeficiency virus (HIV) in Florianópolis, Brazil. A total of 2,678 serum samples from 2,583 blood donors and 95 HIV-infected patients, collected between April, 1994, and March, 1995, were examined for markers of HBV and HCV. All the samples were analyzed to detect HBV and HCV markers (HBsAg, anti-HBc, and anti-HCV). Hepatitis B and C prevalence among the studied blood donors reached 9.3% and 1.0%, respectively; 0.7% being seropositive for HBsAg and 9.2% for anti-HBc. It was also verified that 0.1% of blood donors were seropositive for HBsAg alone, 8.6% seropositive for the anti-HBc alone, and 0.6% presented a positive reaction for both of the HBV markers studied. Among HIV-infected patients, prevalence of 69.5% and 54.7% for hepatitis B and hepatitis C, respectively, were observed. Of these patients, 18.9% were seropositive for HBsAg, and 66.3% for the anti-HBc. The prevalence of a reaction for HBsAg alone, and for anti-HBc alone was 3.1% and 50.5%, respectively, for HIV-infected patients, whereas 15.8% were seropositive for both of the studied markers. HBV and HCV coinfection was 0.1% in blood donors, and 40% of those patients tested seropositive for HIV. Results show prevalence of HBV and HCV infection to be significantly greater among HIV-infected patients than among blood donors. These observations confirm the high frequency of HIV-infected patients exposure to these other viruses.

Utilization of the depressor anguli oris musculocutaneous flap for lip reconstruction.

The authors describe the anatomic aspects and surgical technique of the depressor anguli oris musculocutaneous flap for reconstruction of the upper and lower lips. Twenty patients were submitted to surgical treatment, 19 for carcinoma and for upper lip scar deformity. In all patients the repair was performed with the depressor anguli oris musculocutaneous island flap. At the follow-up, lip function was satisfactory in 19 patients and unsatisfactory in 1 patient. The aesthetic results were considered satisfactory in all patients. The depressor anguli oris musculocutaneous island flap is safe for upper and lower lip reconstruction, with good functional and aesthetic results, and can be added as a new flap for lip reconstruction.

Relationship between the actions of atrial natriuretic peptide (ANP), guanylin and uroguanylin on the isolated kidney.

Guanylin and uroguanylin are peptides that bind to and activate guanylate cyclase C and control salt and water transport in many epithelia in vertebrates, mimicking the action of several heat-stable bacteria enterotoxins. In the kidney, both of them have well-documented natriuretic and kaliuretic effects. Since atrial natriuretic peptide (ANP) also has a natriuretic effect mediated by cGMP, experiments were designed in the isolated perfused rat kidney to identify possible synergisms between ANP, guanylin and uroguanylin. Inulin was added to the perfusate and glomerular filtration rate (GFR) was determined at 10-min intervals. Sodium was also determined. Electrolyte dynamics were measured by the clearance formula. Guanylin (0.5 microg/ml, N = 12) or uroguanylin (0.5 microg/ml, N = 9) was added to the system after 30 min of perfusion with ANP (0.1 ng/ml). The data were compared at 30-min intervals to a control (N = 12) perfused with modified Krebs-Hanseleit solution and to experiments using guanylin and uroguanylin at the same dose (0.5 microg/ml). After previous introduction of ANP in the system, guanylin promoted a reduction in fractional sodium transport (%TNa+, P<0.05) (from 78.46 +/- 0.86 to 64.62 +/- 1.92, 120 min). In contrast, ANP blocked uroguanylin-induced increase in urine flow (from 0.21 +/- 0.01 to 0.15 +/- 0.007 ml g-1 min-1, 120 min, P<0.05) and the reduction in fractional sodium transport (from 72.04 +/- 0. 86 to 85.19 +/- 1.48, %TNa+, at 120 min of perfusion, P<0.05). Thus, the synergism between ANP + guanylin and the antagonism between ANP + uroguanylin indicate the existence of different subtypes of receptors mediating the renal actions of guanylins.

Relationship between the actions of atrial natriuretic peptide (ANP), guanylin and uroguanylin on the isolated kidney

Guanylin and uroguanylin are peptides that bind to and activate guanylate cyclase C and control salt and water transport in many epithelia in vertebrates, mimicking the action of several heat-stable bacteria enterotoxins. In the kidney, both of them have well-documented natriuretic and kaliuretic effects. Since atrial natriuretic peptide (ANP) also has a natriuretic effect mediated by cGMP, experiments were designed in the isolated perfused rat kidney to identify possible synergisms between ANP, guanylin and uroguanylin. Inulin was added to the perfusate and glomerular filtration rate (GFR) was determined at 10-min intervals. Sodium was also determined. Electrolyte dynamics were measured by the clearance formula. Guanylin (0.5 µg/ml, N = 12) or uroguanylin (0.5 µg/ml, N = 9) was added to the system after 30 min of perfusion with ANP (0.1 ng/ml). The data were compared at 30-min intervals to a control (N = 12) perfused with modified Krebs-Hanseleit solution and to experiments using guanylin and uroguanylin at the same dose (0.5 µg/ml). After previous introduction of ANP in the system, guanylin promoted a reduction in fractional sodium transport (TNa+, P<0.05) (from 78.46 + or - 0.86 to 64.62 = or - 1.92, 120 min). In contrast, ANP blocked uroguanylin-induced increase in urine flow (from 0.21 = or - 0.01 to 0.15 + or - 0.007 ml g-1 min-1, 120 min, P<0.05) and the reduction in fractional sodium transport (from 72.04 + or - 0.86 to 85.19 + or - 1.48, TNa+, at 120 min of perfusion, P<0.05). Thus, the synergism between ANP + guanylin and the antagonism between ANP + uroguanylin indicate the existence of different subtypes of receptors mediating the renal actions of guanylins

[Langerhans cell histiocytosis in anogenital region]. / Histiocitose das células de Langerhans na região anogenital.

The Langerhans cell histiocytosis (LCH) of the genital tract is rare, with only 48 cases related in the literature. There were reported only 2 cases in the anogenital region. We reported the third case of LCH in the anogenital region; patient was female, 31 years-old, caucasian and the diagnosis was confirmed by electron microscopic magnification. The treatment was local surgical excision and systemic chemotherapy.

Efficacy of a glutamine-based oral rehydration solution on the electrolyte and water absorption in a rabbit model of secretory diarrhea induced by cholera toxin.

BACKGROUND: Glutamine is absorbed in the intestinal tract coupled with sodium and is the principal metabolic substrate for the enterocyte. Therefore, an oral rehydration solution containing this substance might provide an effective oral means of restoring electrolyte losses as well as speeding repair of mucosal damage. The objective of this work was to investigate the use of an oral rehydration solution based on glutamine in vivo in the perfused rabbit ileal loop model of secretory diarrhea induced by choleratoxin. METHODS: Phenolsulfonphthalein (PSP, 50 mg/l) was used as a nonabsorbable marker for calculations of net water and electrolyte transport. Solutions tested included: (a) a glutamine-based oral rehydration solution with 111 mmol/l glutamine, (Gln-ORS); (b) the oral rehydration solution recommended by the World Health Organization; (c) modified Ringer's solution. Choleratoxin (1 microg/ml) was injected into the lumen of the ileal rabbit segments for 30 minutes prior to the initiation of the perfusion. RESULTS: Choleratoxin induced significant secretion of sodium in the control modified Ringer's solution (10.8 +/- 2.95 vs -14.05 +/- 5.95 microEq/g/min, n = 10; p < 0.01) and of water (0.06 +/- 0.03 vs -0.15 +/- 0.06 ml/g/min, n = 10; p < 0.01) with a maximum effect at 60 minutes after initiation of perfusion. World Health Organization oral rehydration solution was able to significantly reduce the intestinal secretion of sodium (control with cholera = -14.34 +/- 2.18 vs oral rehydration solution with cholera = -0.50 +/- 0.48 microEq/g/min, n = 10; p < 0.01) and water (-0.15 +/- 0.02 vs -0.012 +/- 0.005 ml/g/min, n = 10; p < 0.01). For comparison, glutamine-based oral rehydration solution had an even greater effect on sodium and water absorption (glutamine-based oral rehydration solution with choleratoxin = 10.31 +/- 1.21 microEq/g/min, n = 5; p < 0.01 for sodium and 0.08 +/- 0.008 ml water/g/min; n = 5; p < 0.01). Choleratoxin did not change the effect of glutamine-based oral rehydration solution on sodium and water absorption (12.90 +/- -1.09 microEq sodium/g/min, n = 5; and 0.11 +/- 0.01 ml water/g/min; n = 5). In addition glutamine-based oral rehydration solution also induced a greater absorption of potassium and chloride in the intestinal ileal segments treated with choleratoxin compared with World Health Organization glutamine-based oral rehydration solution. CONCLUSIONS: These results demonstrate the superior efficacy of glutamine-based oral rehydration solution in electrolyte and water absorption compared with modified Ringer's control solution or even with World Health Organization-recommended oral rehydration solution.

The lysine-1 analog of guanylin induces intestinal secretion and natriuresis in the isolated perfused kidney.

Guanylin is an endogenous peptide synthesized by several mammalian species that mimics the effects of a thermostable enterotoxin of Escherichia coli (STa: NTFYCCELCCNPACAGCY) in the gut. We have cloned a lysine-1 derivative of rat guanylin (Lys-1-NTCEICAYAACTGC) and tested its effects on ileal tissue membranes in Ussing chambers and in the isolated perfused rat kidney. Rabbit ileal mucosa membranes were mounted into a Ussing chamber and the effects of Lys-1 guanylin (Lys-1 G) and STa enterotoxin peptide on chloride secretion were determined by changes in short-circuit current (Isc). Lys-1 G (10 to 100 nM) showed a dose-dependent effect on chloride secretion with a maximal response estimated to be 52 microA/cm2. Lys-1 G mimics the effect of STa peptide, but the enterotoxin elicited a greater maximal effect of 120 microA/cm2 (P < 0.01). Lys-1 G (2.5 micrograms/ml) promoted an increase in both urine flow (from 0.13 +/- 0.07 to 0.40 +/- 0.01 ml g-1 min-1, N = 4; P < 0.05) and glomerular filtration rate (from 0.68 +/- 0.02 to 0.85 +/- 0.00 ml g-1 min-1, N = 4; P < 0.01) in the isolated perfused kidney and a reduction of the fractional reabsorption of sodium (from 76.0 +/- 0.03 to 59.5 +/- 0.85%, N = 4; P < 0.01). These maximal effects were accompanied by intense natriuretic effect observed 30 and 60 min after drug administration. The Lys-1 G analog similar to STa enterotoxin elicited intestinal chloride secretion and a natriuretic effect. These data demonstrate that the cloned peptide analog retains the biological activity of the native hormone and presents activity similar to STa. The properties of Lys-1 G resemble those of a factor formed during perfusion of the hypoxic rabbit kidney and named by us factor natriureticus similis (FNS).

The lysine-1 analog of guanylin induces intestinal secretion and natriuresis in the isolated perfused kidney

Guanylin is an endogenous peptide synthesized by several mammalian species that mimics the effects of a thermostable enterotoxin of Escherichia coli (STa: NTFYCCELCCNPACAGCY) in the gut. We have cloned a lysine-1 derivative of rat guanylin (Lys-1-NTCEICAYAACTGC) and tested its effects on ileal tissue membranes in Ussing chambers and in the isolated perfused rat kidney. Rabbit ileal mucosa membranes were mounted into a Ussing chamber and the effects of Lys-1 guanylin (Lys-1 G) and STa enterotoxin peptide on chloride secretion were determined by changes in short-circuit current (Isc). Lys-1 G (10 to 100 nM) showed a dose-dependent effect on chloride secretion with a maximal response estimated to be 52 microA/cm2. Lys-1 G mimics the effect of STa peptide, but the enterotoxin elicited a greater maximal effect of 120 microA/cm2 (p<0.01). Lys-1 G (2.5 microg/ml) promoted an increase in both urine flow (from 0.13 +/- 0.07 to 0.40 +/- 0.01 ml g(-1) min(-1), N = 4; P<0.05) and glomerular filtration rate (from 0.68 +/- 0.02 to 0.85 0.00 ml g(-1) min(-1), N = 4; P<0.01) in the isolated perfused kidney and a reduction of the fractional reabsorption of sodium (from 76.0 +/- 0.03 to 59.5 +/- 0.85 percent, N = 4; P<0.01). These maximal effects were accompanied by intense natriuretic effect observed 30 and 60 min after drug administration. The Lys-1 G analog similar to STa enterotoxin elicited intestinal chloride secretion and a natriuretic effect. These data demonstrate that the cloned peptide analog retains the biological activity of the native hormone and presents activity similar to STa. The properties of Lys-1 G resemble those of a factor formed during perfusion of the hypoxic rabbit kidney and named by us factor natriureticus similis (FNS).

Surgical correction of nipple hypertrophy.

In this report, a surgical technique for reduction of nipple height and diameter without affecting the central column is described. The results are discussed in light of existing literature.

Vascular and glomerular effects of Clostridium difficile toxin A peptide on the isolated rat kidney.

Toxin A peptide from Clostridium difficile caused damage and secretion in the intestinal mucosa. These effects are mediated in part by pro-inflammatory substances. In order to evaluate and compare the biologic effect of toxin A on renal vascular, glomerular and tubular functions, we studied this toxin in isolated rat kidneys. Isolated kidneys from adult male Wistar rats (260-320 g) were perfused with Krebs-Henseleit solution containing 60 mg/ml dialyzed bovine serum albumin. We studied the effect of toxin A peptide (3.2 x 10(-6) M, injected into perfusate) on glomerular filtration rate (GFR), urinary flow rate (UF) and total sodium reabsorption (TNa+, %). All experiments were preceded by a 30-min basal period, and in another group of kidneys the time course of the variables was followed without toxin infusion for unpaired control. Toxin A (TxA) reduced the perfusion pressure (PP), from PPcontrol/30min = 124.89 +/- 1.91 to PPTxA/120min = 88.13 +/- 5.1 mmHg (N = 6, P < 0.01) with a maximal effect at 120 min after toxin infusion. TxA also caused a significant decrease in GFR with maximal effect at 90 min after toxin infusion (GFRcontrol/30min = 0.53 +/- 0.05 to GFRTxA/90min = 0.30 + 0.05 ml min-1g-1; N = 6, P < 0.01). TxA did not alter renal tubular sodium transport when compared with a control without toxin infusion. In addition, toxin-treated kidneys caused a time-dependent increase in urinary flow from UFcontrol/30min = 0.16 +/- 0.08 to UFTxA/120min = 0.35 +/- 0.1 ml min-1g-1 (N = 6, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Vascular and glomerular effects of Clostridium difficile toxin A peptide on the isolated rat kidney

Toxin A peptide from Clostridium difficile caused damage and secretion in the intestinal mucosa. These effects are mediated in part by pro-inflammatory substances. In order to evaluate and compare the biologic effect of toxin A on renal vascular, glomerular and tubular functions, we studied this toxin in isolated rat kidneys. Isolated kidneys from adult male Wistar rats (260-320 g) were perfused with Krebs-Henseleit solution containing 60 mg/ml dialyzed bovine serum albumin. We studied the effect of toxin A peptide (3.2 x 10(-6) M, injected into perfusate) on glomerular filtration rate (GFR), urinary flow rate (UF) and total sodium reabsorption (TNa+, per cent ). All experiments were preceded by a 30-min basal period, and in another group of kidneys the time course of the variables was followed without toxin infusion for unpaired control. Toxin A (TxA) reduced the perfusion pressure (PP), from PPcontrol/30min = 124.89 +/- 1.91 to PPTxA/120min = 88.13 +/- 5.1 mmHg (N = 6, P < 0.01) with a maximal effect at 120 min after toxin infusion. TxA also caused a significant decrease in GFR with maximal effect at 90 min after toxin infusion (GFRcontrol/30min = 0.53 +/- 0.05 to GFRTxA/90min = 0.30 + 0.05 ml min-1g-1; N = 6, P < 0.01). TxA did not alter renal tubular sodium transport when compared with a control without toxin infusion. In addition, toxin-treated kidneys caused a time-dependent increase in urinary flow from UFcontrol/30min = 0.16 +/- 0.08 to UFTxA/120min = 0.35 +/- 0.1 ml min-1g-1 (N = 6, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)