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94 Efficacy of theophylline in promoting ureteral stone passage
93
94
C O M P A R I S O N OF D I C L O F E N A C S O D I U M AND I N T R A C U T A N E STERILE WATER INJECTIONS IN R E N A L COLIC: A R A N D O M I Z E D TRIAL
E F F I C A C Y OF T H E O P H Y L L I N E IN P R O M O T I N G U R E T E R A L STC NE PASSAGE
Sigirci A.R., Seymen T., Karada~ S., Aras B., Sahin S., 0zbay B., Ba~ M., Kemahh E., Tfl~91A.I
Tadavvon F., Shafiei L.
Bakirk6y. Dr. Sadi Konuk Research and Training Hospital, Department of Urology, Istanbul, Turkey
Noor Hospital, Esfahan Medical Science University, Esfahan, Iran
I N T R O D U C T I O N & OBJECTIVES: We evaluate the pain relief in renal colic with Diclofenac Sodium intramuscularly and intracutane sterile water injections, compared with placebo (NaCI 0.9%).
I N T R O D U C T I O N & OBJECTIVES: As we know ureteral stones are a very
M A T E R I A L & METHODS: 119 patients with renal colic, who did not receive any analgesic therapy 6 hours prior, were investigated. They were randomly assessed into 3 groups. 39 of them received 75 mg Diclofenac Sodium intramuscularly (i.m.) (Group 1); 40 received intracutane 4x0.2 ml sterile water at the trigger points of the affected flank (Group 2) wherein the 3rd group (placebo) received 4x0.2 ml of saline (0.9% NaC1) intracutaneous at the same points. The pain relief was measured using the visual analogue scale (VAS) before treatment and at minutes 1, 5, 15, 30, 60, 120, and 360 after the injections.
common problem. Theophyllin causes smooth muscle relaxation by increa~;ing cAMP. This study was performed to determine the efficacy of this effect of theophyllin in promoting ureteral stone passage.
M A T E R I A L & METHODS: In this study 300 patients (from 17 to 67 years aid) that their ureteral stones were diagnosed by spiral CTscan divided in two equal groups. Stones size were from 2ram to 1lmm. Two groups were matched for age,
RESULTS: The pain scores in all groups measured with the median visual analogue scale before treatment were similar. (Group 1:8.7 cm; Group 2:8.8 cm and Group 3 : 8 . 8 cm). But starting from 1st minute in all checking minutes (incl. 360.rain) there were (except rain. 120) significantly differences in favour of the sterile water group :lst minute 8.8 cm vs. 5.7 cm (p<0.01); 5th rain. 8.3 cm vs. 2.1 cm (p<0.001); 15th min. 6.0 cm vs. 2.1 cm (p<0.001); 30th min. 3.0 cm vs. 1.1 cm (p<0.001); 60.th rain 1.1 cm vs. 0.5 cm (!o<0.001); 120.th rain. 0.7 cm vs 0.6 cm (p>0.5) and 360. min. 1.2 cm vs. 0.6 cm p<0.001). Wherein in the placebo group (Group 3) only in the 1. minute with 4.5 cm a reduction in pain was observed; the pain expressed in VAS was 5.1 cm; in min. 5; 6.2cm in rain.15 and 6.9 in minute 30. We stopped to evaluate these patients due to ethical reasons after the 30. minute and applied Diclofenac Sodium 75 mg im (n=20) or intracutane sterile water (n=20). CONCLUSIONS: The analgesic effect of intracutaneous applied sterile water is superior to the classical therapy modality with nonsteroidal anti inflammatory drugs (NSAID) in amount to rapid and long acting pain relief in renal colic. The placebo group showed except in the initial period no effect in pain relief in renal colic.
s e x , location of stone in ureter and stone size. The amount of liquid intake was similar in two groups. Group A received oral theophyllin (200 mg bid) and group B received placebo, both groups for 6 weeks. At the end of 6 weeks stone pas,;age was determined by spiral CTscan.
RESULTS: In our study 92 patients in group A (61.3%) and 62 patients in group B (41.3%) passed their stones.(p<0.032) The average time for stone passage was 18.3 + /
2.4 days in group A and
24.8+/3.1 days in group B. (p<0.05)
CONCLUSIONS: Our finding demonstrate the positive effect of theophyllin in promoting ureteral stone passage.
95
96
EVALUATION OF CYSTINE T R A N S P O R T IN C U L T U R E D H U M A N KIDNEY CELLS: P R I M E R TO GENE T H E R A P Y OF CYSTINURIA
DOSAGE ALTERATIONS TO PREVENT I N T R A T U B U L A R INDIN~VIR CRYSTALLIZATION
Sagi S., Trojan L., Alken P., Michel M.S., Knoll Y.
Salahuddin S}, Kok D. 2, Buchholz N. 3
Mannheim University Hospital, Department of Urology, Mannheim, Germany
'Eastboume General Hospital, General Practice, Eastbourne, United Kingdom, ZErasmus University, Experimental Urology, Rotterdam, The Netherlands, 3Barts and the London NHS Trust, Urology, London, United Kingdom
INTRODUCTION & OBJECTIVES: Cystinnria is a rare hereditary disease resulting in recurrent stone formation and the need for repeated invasive interventions. It is the cause of 1-2% of stones observed in adults and about 10% of those occurring in children. Two responsible genes have been identified which encode the transporters rBAT and b°,+AT to form a beterodimer and transport cystine and leucine among others and whose defect results in crystallisation of cystine. Aim of this study was to establish the presence of cystine transport mechanism in human PTCs when cultured in vitro and to generate a cystinuria phenotype in the above cells. MATERIAL & METHODS: HK-2 is a proximal tubular cell (PTC) line derived from normal human kidney. The presence of rBAT gene was determined using the c-DNA of HK-2 cells by PCR with the rBAT primers. Total protein from the HK-2 cells was isolated. Western blot using peptide antibodies generated against the protein rBAT was done to check for the gene expression. Radioactively labeled amino acids cystine (S35) and leucine (H 3) were used to determine the functional presence of this amino acid transport in HK-2 cells when cultured in vitro. To achieve a cystinuria phenotype in HK2 cells, the rBAT gene was silenced using the antisense oligonucleotides. RESULTS: The PCR product (170 bp) of the c-DNA verified the presence of the rBAT gene: Similarly the western blot at 87 kD shows the expression of the rBAT gene. Cellular uptake with lgCi of cystine per well for an incubation time of 5 rain was around 0.05%. 0.18% uptake was observed for 15 minutes of incubation. For an incubation time of 30 rain, the uptake was around 1% for cystine. In comparison, cellular uptake for lgCi of leucine was 1.9%, 3.2% and 4.3% for 5, 15 and 30 minutes of incubation. A transient knock out of the rBAT gene was seen with the delivery of antisense oligos. There was an uptake of 0.75% of cystine with the suppression of the transport by antisense oligos in comparison to approximately 1% of cystine uptake without the antisense oligos. CONCLUSIONS: This study demonstrates the existence of the ba;+ amino acid transport system in normal human proximal tubular HK-2 cells when cultured in vitro and it shows that it is possible to efficiently suppress this b°;+ transport system leading to a state comparable to human cystinuria. With this knowledge it will be possible to establish for the first time an in vitro model of cystinuria in cultured human kidney proximal tubule cells.
European Urology Supplements 4 (2005) No. 3, pp. 26
INTRODUCTION & OBJECTIVES: The authors have previously shown that indinavir (IND) is likely to precipitate in the loop of Henl6 (LH) at plasma concentrations of 8 mg/L. This resulted in recommendations of drug dosage adjustments. However, those experiments were performed at room temperature. Given the influence of temperature on crystallisation in general, and solubility of 1ND in particular, we repeated the experiments under physiological body temperature conditions. MATERIAL & METHODS: Test solutions contained IND concentrations (CINI)) of 100-750 mg/L at ionic strengths (IS) varying from 0-800mmol simulating conditions in the proximal tubule (PT) and the LH. Solutions were titrated with base (NaOH) to find the pH value where nucleation starts within one minute. Each measurement was repeated three-fold and the average values were used for analysis. Experiments were conducted at room temperature (20°C) and repeated under a constantly monitored temperature of 37°C. RESULTS: Results at 20°C were similar to our previous experiments. At body temperature, the relationship between the pH and CIND remained inversely proportional. Again, results confirmed the localisation oflND crystallisation to be in the LH. Precipitation occurred already at lower CIND of 100 mg/L in the LH, as compared to 125 mg/L at 20°C, corresponding to a lower plasma CIND of 6.41 mg/L, as compared to 8.01 mg/L at 20°C. Precipitation at 37°C occurred at lower pH-values (6.67-7.26 versus 7.23-7.44). Precipitation under PT conditions corresponded to 64.1 mg/L plasma concentration, which cannot be reached in vivo under current dosage regimens. CONCLUSIONS: This study confirms that the most likely place for IND crystallisation is the LH. It precipitates at lower concentrations and lower pH at body temperature. Therefore, the corresponding plasma concentration leading to 1ND crystallisation and consecutive stone formation is even lower than previously reported by us. We therefore like to emphasize the importance of adjusting IND dosage regimens in a way that avoids excessive peak values yet remains well above the effective plasma CIND. Since precipitation at 37°C occurred at very low CIND, a rest-risk of '.stone formation cannot be excluded completely even in patients where this adjustment has been achieved.
94
C O M P A R I S O N OF D I C L O F E N A C S O D I U M AND I N T R A C U T A N E STERILE WATER INJECTIONS IN R E N A L COLIC: A R A N D O M I Z E D TRIAL
E F F I C A C Y OF T H E O P H Y L L I N E IN P R O M O T I N G U R E T E R A L STC NE PASSAGE
Sigirci A.R., Seymen T., Karada~ S., Aras B., Sahin S., 0zbay B., Ba~ M., Kemahh E., Tfl~91A.I
Tadavvon F., Shafiei L.
Bakirk6y. Dr. Sadi Konuk Research and Training Hospital, Department of Urology, Istanbul, Turkey
Noor Hospital, Esfahan Medical Science University, Esfahan, Iran
I N T R O D U C T I O N & OBJECTIVES: We evaluate the pain relief in renal colic with Diclofenac Sodium intramuscularly and intracutane sterile water injections, compared with placebo (NaCI 0.9%).
I N T R O D U C T I O N & OBJECTIVES: As we know ureteral stones are a very
M A T E R I A L & METHODS: 119 patients with renal colic, who did not receive any analgesic therapy 6 hours prior, were investigated. They were randomly assessed into 3 groups. 39 of them received 75 mg Diclofenac Sodium intramuscularly (i.m.) (Group 1); 40 received intracutane 4x0.2 ml sterile water at the trigger points of the affected flank (Group 2) wherein the 3rd group (placebo) received 4x0.2 ml of saline (0.9% NaC1) intracutaneous at the same points. The pain relief was measured using the visual analogue scale (VAS) before treatment and at minutes 1, 5, 15, 30, 60, 120, and 360 after the injections.
common problem. Theophyllin causes smooth muscle relaxation by increa~;ing cAMP. This study was performed to determine the efficacy of this effect of theophyllin in promoting ureteral stone passage.
M A T E R I A L & METHODS: In this study 300 patients (from 17 to 67 years aid) that their ureteral stones were diagnosed by spiral CTscan divided in two equal groups. Stones size were from 2ram to 1lmm. Two groups were matched for age,
RESULTS: The pain scores in all groups measured with the median visual analogue scale before treatment were similar. (Group 1:8.7 cm; Group 2:8.8 cm and Group 3 : 8 . 8 cm). But starting from 1st minute in all checking minutes (incl. 360.rain) there were (except rain. 120) significantly differences in favour of the sterile water group :lst minute 8.8 cm vs. 5.7 cm (p<0.01); 5th rain. 8.3 cm vs. 2.1 cm (p<0.001); 15th min. 6.0 cm vs. 2.1 cm (p<0.001); 30th min. 3.0 cm vs. 1.1 cm (p<0.001); 60.th rain 1.1 cm vs. 0.5 cm (!o<0.001); 120.th rain. 0.7 cm vs 0.6 cm (p>0.5) and 360. min. 1.2 cm vs. 0.6 cm p<0.001). Wherein in the placebo group (Group 3) only in the 1. minute with 4.5 cm a reduction in pain was observed; the pain expressed in VAS was 5.1 cm; in min. 5; 6.2cm in rain.15 and 6.9 in minute 30. We stopped to evaluate these patients due to ethical reasons after the 30. minute and applied Diclofenac Sodium 75 mg im (n=20) or intracutane sterile water (n=20). CONCLUSIONS: The analgesic effect of intracutaneous applied sterile water is superior to the classical therapy modality with nonsteroidal anti inflammatory drugs (NSAID) in amount to rapid and long acting pain relief in renal colic. The placebo group showed except in the initial period no effect in pain relief in renal colic.
s e x , location of stone in ureter and stone size. The amount of liquid intake was similar in two groups. Group A received oral theophyllin (200 mg bid) and group B received placebo, both groups for 6 weeks. At the end of 6 weeks stone pas,;age was determined by spiral CTscan.
RESULTS: In our study 92 patients in group A (61.3%) and 62 patients in group B (41.3%) passed their stones.(p<0.032) The average time for stone passage was 18.3 + /
2.4 days in group A and
24.8+/3.1 days in group B. (p<0.05)
CONCLUSIONS: Our finding demonstrate the positive effect of theophyllin in promoting ureteral stone passage.
95
96
EVALUATION OF CYSTINE T R A N S P O R T IN C U L T U R E D H U M A N KIDNEY CELLS: P R I M E R TO GENE T H E R A P Y OF CYSTINURIA
DOSAGE ALTERATIONS TO PREVENT I N T R A T U B U L A R INDIN~VIR CRYSTALLIZATION
Sagi S., Trojan L., Alken P., Michel M.S., Knoll Y.
Salahuddin S}, Kok D. 2, Buchholz N. 3
Mannheim University Hospital, Department of Urology, Mannheim, Germany
'Eastboume General Hospital, General Practice, Eastbourne, United Kingdom, ZErasmus University, Experimental Urology, Rotterdam, The Netherlands, 3Barts and the London NHS Trust, Urology, London, United Kingdom
INTRODUCTION & OBJECTIVES: Cystinnria is a rare hereditary disease resulting in recurrent stone formation and the need for repeated invasive interventions. It is the cause of 1-2% of stones observed in adults and about 10% of those occurring in children. Two responsible genes have been identified which encode the transporters rBAT and b°,+AT to form a beterodimer and transport cystine and leucine among others and whose defect results in crystallisation of cystine. Aim of this study was to establish the presence of cystine transport mechanism in human PTCs when cultured in vitro and to generate a cystinuria phenotype in the above cells. MATERIAL & METHODS: HK-2 is a proximal tubular cell (PTC) line derived from normal human kidney. The presence of rBAT gene was determined using the c-DNA of HK-2 cells by PCR with the rBAT primers. Total protein from the HK-2 cells was isolated. Western blot using peptide antibodies generated against the protein rBAT was done to check for the gene expression. Radioactively labeled amino acids cystine (S35) and leucine (H 3) were used to determine the functional presence of this amino acid transport in HK-2 cells when cultured in vitro. To achieve a cystinuria phenotype in HK2 cells, the rBAT gene was silenced using the antisense oligonucleotides. RESULTS: The PCR product (170 bp) of the c-DNA verified the presence of the rBAT gene: Similarly the western blot at 87 kD shows the expression of the rBAT gene. Cellular uptake with lgCi of cystine per well for an incubation time of 5 rain was around 0.05%. 0.18% uptake was observed for 15 minutes of incubation. For an incubation time of 30 rain, the uptake was around 1% for cystine. In comparison, cellular uptake for lgCi of leucine was 1.9%, 3.2% and 4.3% for 5, 15 and 30 minutes of incubation. A transient knock out of the rBAT gene was seen with the delivery of antisense oligos. There was an uptake of 0.75% of cystine with the suppression of the transport by antisense oligos in comparison to approximately 1% of cystine uptake without the antisense oligos. CONCLUSIONS: This study demonstrates the existence of the ba;+ amino acid transport system in normal human proximal tubular HK-2 cells when cultured in vitro and it shows that it is possible to efficiently suppress this b°;+ transport system leading to a state comparable to human cystinuria. With this knowledge it will be possible to establish for the first time an in vitro model of cystinuria in cultured human kidney proximal tubule cells.
European Urology Supplements 4 (2005) No. 3, pp. 26
INTRODUCTION & OBJECTIVES: The authors have previously shown that indinavir (IND) is likely to precipitate in the loop of Henl6 (LH) at plasma concentrations of 8 mg/L. This resulted in recommendations of drug dosage adjustments. However, those experiments were performed at room temperature. Given the influence of temperature on crystallisation in general, and solubility of 1ND in particular, we repeated the experiments under physiological body temperature conditions. MATERIAL & METHODS: Test solutions contained IND concentrations (CINI)) of 100-750 mg/L at ionic strengths (IS) varying from 0-800mmol simulating conditions in the proximal tubule (PT) and the LH. Solutions were titrated with base (NaOH) to find the pH value where nucleation starts within one minute. Each measurement was repeated three-fold and the average values were used for analysis. Experiments were conducted at room temperature (20°C) and repeated under a constantly monitored temperature of 37°C. RESULTS: Results at 20°C were similar to our previous experiments. At body temperature, the relationship between the pH and CIND remained inversely proportional. Again, results confirmed the localisation oflND crystallisation to be in the LH. Precipitation occurred already at lower CIND of 100 mg/L in the LH, as compared to 125 mg/L at 20°C, corresponding to a lower plasma CIND of 6.41 mg/L, as compared to 8.01 mg/L at 20°C. Precipitation at 37°C occurred at lower pH-values (6.67-7.26 versus 7.23-7.44). Precipitation under PT conditions corresponded to 64.1 mg/L plasma concentration, which cannot be reached in vivo under current dosage regimens. CONCLUSIONS: This study confirms that the most likely place for IND crystallisation is the LH. It precipitates at lower concentrations and lower pH at body temperature. Therefore, the corresponding plasma concentration leading to 1ND crystallisation and consecutive stone formation is even lower than previously reported by us. We therefore like to emphasize the importance of adjusting IND dosage regimens in a way that avoids excessive peak values yet remains well above the effective plasma CIND. Since precipitation at 37°C occurred at very low CIND, a rest-risk of '.stone formation cannot be excluded completely even in patients where this adjustment has been achieved.