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1.
Actas Urol Esp (Engl Ed) ; 48(5): 364-370, 2024 Jun.
Article in English, Spanish | MEDLINE | ID: mdl-38191025

ABSTRACT

INTRODUCTION AND OBJECTIVE: The implementation of Enhanced Recover After Surgery (ERAS) multimodal rehabilitation protocols in radical cystectomy has shown to improve outcomes in hospital stay and complications. The aim of this analysis is to evaluate the impact of laparoscopic surgery on radical cystectomy within a multimodal rehabilitation program. MATERIAL AND METHODS: The study was carried out in a third level center between 2011 and 2020 including patients with bladder cancer submitted to radical cystectomy according to an ERAS (Enhanced Recovery After Surgery) protocol and the Spanish Multimodal Rehabilitation Group (GERM) with 20 items to be fulfilled. RESULTS: A total of 250 radical cystectomies were performed throughout the study period, 42.8% by open surgery (OS) and 57.2% by laparoscopic surgery (LS). The groups are comparable in demographic and clinical variables (p > 0.05). Operative time was longer in the LS group (248.4 ±â€¯55.0 vs. 286.2 ±â€¯51.9 min; p < 0.001). However, bleeding was significantly lower in the LS group (417.5 ±â€¯365.7 vs. 877.9 ±â€¯529.7 cc; p < 0.001), as was the need for blood transfusion (33.6% vs. 58.9%; p < 0.001). Postoperative length of stay (11.5 ±â€¯10.5 vs. 20.1 ±â€¯17.2 days; p < 0.001), total and major complications were also significantly lower in this group (LS). The readmission rate was lower in the LS group but not significantly (36.4% vs. 29.4%; p = 0.237). The difference between 90-day mortality in both groups was not statistically significant (2.8% LS vs. 4.3% OS; p = 0.546). The differences were maintained in the multivariate models. CONCLUSIONS: Laparoscopic surgery within a multimodal rehabilitation program increases operative time but significantly decreases intraoperative bleeding, transfusion requirements, postoperative length of stay, and complications.


Subject(s)
Cystectomy , Laparoscopy , Urinary Bladder Neoplasms , Humans , Cystectomy/rehabilitation , Cystectomy/methods , Male , Laparoscopy/rehabilitation , Female , Urinary Bladder Neoplasms/surgery , Urinary Bladder Neoplasms/rehabilitation , Aged , Treatment Outcome , Middle Aged , Enhanced Recovery After Surgery , Retrospective Studies , Postoperative Complications/epidemiology , Clinical Protocols , Length of Stay/statistics & numerical data , Combined Modality Therapy
2.
Actas urol. esp ; 45(4): 247-256, mayo 2021. tab
Article in Spanish | IBECS | ID: ibc-216929

ABSTRACT

Introducción y objetivos: La cistectomía radical con derivación urinaria asociada a linfadenectomía pélvica ampliada continúa siendo el tratamiento de elección en el cáncer vesical musculoinvasivo. Un 64% de los pacientes presentan complicaciones postoperatorias, siendo la infección urinaria responsable en un 20-40% de los casos. El objetivo del presente proyecto es valorar la tasa de infección urinaria como causa de reingreso tras cistectomía, e identificar factores protectores y predisponentes de infección urinaria en nuestro medio. Por último, conocer los resultados obtenidos al aplicar el protocolo de profilaxis antibiótica tras la retirada de los catéteres ureterales.Material y métodosEstudio descriptivo retrospectivo de pacientes cistectomizados en el Servicio de Urología del Hospital Clínico Universitario desde enero de 2012 hasta diciembre de 2018. Desde octubre de 2017, de forma estandarizada, a todo paciente se le aplica un protocolo de prevención de infección del tracto urinario (ITU) tras la retirada de catéteres.ResultadosLa ITU es responsable del 54,7% de los reingresos, siendo un 55,1% de estos por causa de una ITU tras la retirada de los catéteres ureterales. El 9,5% de los pacientes con profilaxis presenta ITU tras la retirada, frente a un 10,6% en el grupo de pacientes sin profilaxis. El paciente que reingresa por ITU tras la retirada tiene un tiempo de catéteres medio de 24,3±7,2 días, frente a los 24,5±7,4 días en el grupo sin ITU (p=0,847).ConclusionesEl tipo de derivación urinaria empleada no guarda relación con la tasa de infección urinaria. El modelo de regresión no identifica la profilaxis antibiótica, ni tampoco el tiempo de catéteres, como factores independientes de ITU tras la retirada de los catéteres. (AU)


Introduction and objectives: Radical cystectomy with urinary diversion associated with extended pelvic lymphadenectomy continues to be the treatment of choice in muscle invasive bladder cancer. Sixty-four percent of patients submitted to this procedure present postoperative complications, with urinary infection being responsible in 20-40% of cases. The aim of this project is to assess the rate of urinary infection as a cause of re-admission after cystectomy, and to identify protective and predisposing factors for urinary infection in our environment. Finally, we will evaluate the outcomes after the establishment of a prophylactic antibiotic protocol after removal of ureteral catheters.Material and methodsRetrospective descriptive study of cystectomized patients in the Urology Service of the Hospital Clínico Universitario of Zaragoza, from January 2012 to December 2018. A urinary tract infection (UTI) prevention protocol after catheter removal is established for all patients since October 2017.ResultsUTI is responsible for 54.7% of readmissions, with 55.1% of these being due to UTI after removal of ureteral catheters. Of the patients who received with prophylaxis, 9.5% presented UTIs after withdrawal, compared to 10.6% in the group of patients without prophylaxis. The patient who is re-admitted for UTI after withdrawal has a mean catheter time of 24.3±7.2 days, compared to 24.5±7.4 days for patients in the group without UTI (P=.847).ConclusionsThe type of urinary diversion performed is not related to the rate of urinary infection. The regression model does not identify antibiotic prophylaxis, nor catheter time, as independent factors of UTI after catheter removal. (AU)


Subject(s)
Humans , Antibiotic Prophylaxis , Cystectomy/adverse effects , Urinary Diversion/adverse effects , Urinary Tract Infections/epidemiology , Retrospective Studies
3.
Actas Urol Esp (Engl Ed) ; 45(4): 247-256, 2021 05.
Article in English, Spanish | MEDLINE | ID: mdl-33516599

ABSTRACT

INTRODUCTION AND OBJECTIVES: Radical cystectomy with urinary diversion associated with extended pelvic lymphadenectomy continues to be the treatment of choice in muscle invasive bladder cancer. Sixty-four percent of patients submitted to this procedure present postoperative complications, with urinary infection being responsible in 20-40% of cases. The aim of this project is to assess the rate of urinary infection as a cause of re-admission after cystectomy, and to identify protective and predisposing factors for urinary infection in our environment. Finally, we will evaluate the outcomes after the establishment of a prophylactic antibiotic protocol after removal of ureteral catheters. MATERIAL AND METHODS: Retrospective descriptive study of cystectomized patients in the Urology Service of the Hospital Clínico Universitario of Zaragoza, from January 2012 to December 2018. A urinary tract infection (UTI) prevention protocol after catheter removal is established for all patients since October 2017. RESULTS: UTI is responsible for 54.7% of readmissions, with 55.1% of these being due to UTI after removal of ureteral catheters. Of the patients who received with prophylaxis, 9.5% presented UTIs after withdrawal, compared to 10.6% in the group of patients without prophylaxis. The patient who is re-admitted for UTI after withdrawal has a mean catheter time of 24.3±7.2 days, compared to 24.5±7.4 days for patients in the group without UTI (P=.847). CONCLUSIONS: The type of urinary diversion performed is not related to the rate of urinary infection. The regression model does not identify antibiotic prophylaxis, nor catheter time, as independent factors of UTI after catheter removal.


Subject(s)
Urinary Diversion , Urinary Tract Infections , Antibiotic Prophylaxis , Cystectomy/adverse effects , Humans , Retrospective Studies , Urinary Diversion/adverse effects , Urinary Tract Infections/epidemiology
5.
Cir. Esp. (Ed. impr.) ; 89(10): 663-669, dic. 2011. tab
Article in Spanish | IBECS | ID: ibc-96008

ABSTRACT

Introducción Dada la disponibilidad del abordaje laparoscópico y la creciente detección de incidentalomas, las indicaciones de suprarrenalectomía pueden estar cambiando. La Sección de Cirugía Endocrina de la Asociación Española de Cirujanos diseñó una encuesta para analizar las indicaciones actuales de la suprarrenalectomía y sus resultados en los servicios de cirugía españoles. Material y métodos Se recogieron datos respecto al tipo de hospital y servicio, volumen de procedimientos, estudios de localización y preparación preoperatoria, indicaciones, técnica quirúrgica, instrumental utilizado, y resultados en términos de morbilidad y estancia hospitalaria. Se compararon los resultados de los centros en función de su volumen de actividad, utilizando las pruebas de Mann-Whitney para variables cuantitativas y de la chi al cuadrado para las cualitativas. Resultados Treinta y seis centros cumplimentaron el cuestionario e incluyeron 301 suprarrenalectomías realizadas en 2008. Las lesiones extirpadas más frecuentes fueron feocromocitoma (25,2%), adenoma no funcionante (16,2%), aldosteronoma (15,9%), adenoma de Cushing (11,2%), metástasis (10,3%), mielolipoma (5,6%) y carcinoma (4,9%) (..) (AU)


Introduction The indications for adrenalectomy could be changing by the availability of laparoscopy and the growing detection of incidentalomas. The Endocrine Surgery Section of the Spanish Association of Surgeons conducted a survey to analyse the current indications for adrenalectomy and their results in Spanish surgical departments.Material and methods Data were gathered as regards the type of hospital and department, volume of procedures, localisation studies and pre-surgical preparations, indications, surgical approach, and results in terms of morbidity and hospital stay. The results of the centres were compared as regards their volume of activity using the Mann–Whitney Test for the quantitative variables and chi squared for the qualitative ones. Results Thirty-six centres completed the questionnaire and 301 adrenalectomies were reported to be performed in 2008. Most frequent indications were pheochromocytoma (25.2%), non-functioning adenoma (16.2%), aldosteronoma (15.9%), Cushing adenoma (11.2%), metastasis (10.3%), myelolipoma (5.6%), and carcinoma (4.9%).Laparoscopic adrenalectomy was performed in 83.7% of cases (6.7% required conversion to laparotomy). The mean hospital stay was 3.9 days for laparoscopic adrenalectomy and 7.4 days for laparotomy. High-volume units (more than 10 per year) used more frequently the laparoscopic approach (P=.019), and had a shorter overall hospital stay (P<.0001. Laparoscopic adrenalectomy was also associated with a shorter hospital stay (P<.0001).Conclusions Laparoscopy for adrenalectomy has become the standard practice in Spain, with good results in terms of morbidity and hospital stay. High volume centres have better results as regards the use of minimally invasive surgery and hospital stay (..) (AU)


Subject(s)
Humans , Adrenal Gland Diseases/surgery , Laparoscopy/methods , Adrenalectomy/methods , Postoperative Complications/epidemiology
6.
Actas Urol Esp ; 31(4): 420-2, 2007 Apr.
Article in Spanish | MEDLINE | ID: mdl-17633931

ABSTRACT

In this review we try to update the knowledge about the tumors of epididymis, describing problems in diagnosis and treatment. We present a case of a 39 years old patient who consults by left testicular mass, before the sonogarphy suspect of tumor was made magnetic resonance imaging , wich aimed towards tumorlike injury. Excision of the injury via inguinal was made and the pathologic diagnosis was of adenomatoid tumor. Owing to the few series that appear in literature, and being the commentaries of these tumors about isolated cases, we expose the characteristics of this illustrated case to value the characteristics in diagnosis and treatment to compare them with other cases.


Subject(s)
Adenomatoid Tumor/diagnosis , Epididymis , Genital Neoplasms, Male/diagnosis , Scrotum , Adult , Humans , Male
7.
Actas urol. esp ; 31(4): 420-422, abr. 2007. ilus
Article in Es | IBECS | ID: ibc-054101

ABSTRACT

Con esta revisión pretendemos actualizar los conocimientos acerca de los tumores de epidídimo, describiendo problemas en el diagnostico y tratamiento. Presentamos el caso de un paciente de 39 años que consulta por masa testicular izquierda, ante la duda ecográfica de tumor se realizó resonancia nuclear magnética que apuntaba hacia lesión tumoral. Se realizó exéresis de la lesión vía inguinal y el diagnóstico anatomopatológico fue de tumor adenomatoide. Dada las escasas series que aparecen en la literatura, y siendo los comentarios de estos tumores acerca de casos aislados, creemos oportuno exponer las características de este caso ilustrado iconográficamente para valorar las características diagnosticas y actitud terapéutica para poder compararlas con otros casos


In this review we try to update the knowledge about the tumors of epididymis, describing problems in diagnosis and treatment. We present a case of a 39 years old patient who consults by left testicular mass, before the sonogarphy suspect of tumor was made magnetic resonance imaging , wich aimed towards tumorlike injury. Excision of the injury via inguinal was made and the pathologic diagnosis was of adenomatoid tumor. Owing to the few series that appear in literature, and being the commentaries of these tumors about isolated cases, we expose the characteristics of this illustrated case to value the characteristics in diagnosis and treatment to compare them with other cases


Subject(s)
Male , Humans , Epididymis/pathology , Testicular Neoplasms/pathology , Adenomatoid Tumor/pathology , Magnetic Resonance Spectroscopy
8.
Article in English | MEDLINE | ID: mdl-15876224

ABSTRACT

The in vitro susceptibility to penicillin G, erythromycin and clindamycin was determined by the disc diffusion test and by E-test for a total of 47 streptococcal strains (three Streptococcus uberis, 36 Streptococcus agalactiae, eight Streptococcus dysgalactiae spp. dysgalactiae) isolated from bovine intramammary infections in Argentina. Moreover, resistance phenotypes of erythromycin-resistant streptococcal isolates was characterized. MIC90 of penicillin G, erythromycin and clindamycin for S. agalactiae were 0.75, 8.0 and 12.0 microg/ml respectively. Resistance to erythromycin and clindamycin was detected in 13 (27.6%) and 12 (25.5%) isolates respectively. No isolate was resistant to penicillin G. Resistance against macrolides, lincosamides and streptogramin B (MLS(B)) represented by the constitutive MLS(B) phenotype was present in 11 (23.4%) erythromycin-resistant isolates and two isolates (4.3%) expressed the M phenotype. The inducible MLS(B) phenotype was not identified. Results suggest that beta-lactams are the first-line antibiotics when treating streptococcal udder infections; however, the continuous monitoring of the antibiotic resistance is essential, as the emergence of resistant strains has become a growing concern on the therapy of bovine mastitis.


Subject(s)
Anti-Bacterial Agents/pharmacology , Mastitis, Bovine/drug therapy , Streptococcal Infections/veterinary , Streptococcus/drug effects , Animals , Anti-Bacterial Agents/therapeutic use , Argentina , Cattle , Clindamycin/pharmacology , Colony Count, Microbial , Dose-Response Relationship, Drug , Drug Resistance, Bacterial , Drug Resistance, Multiple, Bacterial , Erythromycin/pharmacology , Female , Mastitis, Bovine/microbiology , Microbial Sensitivity Tests/veterinary , Penicillin G/pharmacology , Streptococcal Infections/drug therapy , Streptococcal Infections/microbiology
9.
Rev Argent Microbiol ; 33(2): 108-12, 2001.
Article in Spanish | MEDLINE | ID: mdl-11494753

ABSTRACT

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.


Subject(s)
Antigenic Variation , Antigens, Bacterial/immunology , Cattle Diseases/microbiology , Leptospira interrogans/immunology , Weil Disease/veterinary , Agglutination Tests , Animals , Antibodies, Bacterial/blood , Antibodies, Bacterial/immunology , Antibody Specificity , Antigens, Bacterial/isolation & purification , Argentina , Bacterial Proteins/immunology , Bacterial Proteins/isolation & purification , Bacterial Vaccines , Blotting, Western , Cattle , Cattle Diseases/blood , Electrophoresis, Polyacrylamide Gel , Leptospira interrogans/classification , Leptospira interrogans/isolation & purification , Reference Standards , Staining and Labeling , Weil Disease/blood , Weil Disease/microbiology
10.
Rev. argent. microbiol ; 33(2): 108-112, abr.-jun. 2001.
Article in Spanish | BINACIS | ID: bin-6760

ABSTRACT

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.(AU)


Subject(s)
Animals , Cattle , Comparative Study , RESEARCH SUPPORT, NON-U.S. GOVT , Antigenic Variation , Antigens, Bacterial/immunology , Cattle Diseases/microbiology , Leptospira interrogans/immunology , Weil Disease/veterinary , Agglutination Tests , Antibodies, Bacterial/blood , Antibodies, Bacterial/immunology , Antibody Specificity , Antigens, Bacterial/isolation & purification , Argentina , Bacterial Proteins/immunology , Bacterial Proteins/isolation & purification , Bacterial Vaccines , Blotting, Western , Cattle Diseases/blood , Electrophoresis, Polyacrylamide Gel , Leptospira interrogans/classification , Leptospira interrogans/isolation & purification , Reference Standards , Staining and Labeling , Weil Disease/blood , Weil Disease/microbiology
11.
Rev. argent. microbiol ; 33(2): 108-112, abr.-jun. 2001.
Article in Spanish | LILACS | ID: lil-332493

ABSTRACT

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.


Subject(s)
Animals , Cattle , Antigenic Variation , Antigens, Bacterial/immunology , Weil Disease/veterinary , Cattle Diseases/microbiology , Leptospira interrogans , Agglutination Tests , Antibody Specificity , Antibodies, Bacterial/blood , Antibodies, Bacterial/immunology , Antigens, Bacterial/isolation & purification , Argentina , Bacterial Vaccines , Blotting, Western , Weil Disease/blood , Weil Disease/microbiology , Cattle Diseases/blood , Electrophoresis, Polyacrylamide Gel , Leptospira interrogans , Bacterial Proteins/immunology , Bacterial Proteins/isolation & purification , Reference Standards , Staining and Labeling
12.
Rev. argent. microbiol ; 33(2): 108-12, 2001 Apr-Jun.
Article in Spanish | BINACIS | ID: bin-39469

ABSTRACT

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.

13.
J Dairy Sci ; 83(6): 1224-7, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10877387

ABSTRACT

A total of 206 strains of Staphylococcus aureus isolated from bovine clinical and subclinical mastitis in Argentina during 1996 to 1998 were investigated for their in vitro susceptibility to several antimicrobial agents. Minimum inhibitory concentrations that inhibit 90% of the strains tested reported in micrograms per milliliters were: 1.5, 0.5, 0.75, 1.50, 0.75, 1.0 and 0.125 for penicillin, oxacillin, cephalothin, gentamicin, erythromycin, clindamycin and ampicillin-sulbactam, respectively. Resistance was detected in 83 (40.3%), 24 (11.6%), 16 (7.7%) and 7 (3.4%) S. aureus isolates for penicillin, erythromycin, pirlimycin and gentamicin, respectively. No resistance was detected for oxacillin, cephalothin and ampicillin-sulbactam. Results indicated that S. aureus isolates in Argentina exhibited high resistance to penicillin of all antimicrobial agents tested.


Subject(s)
Anti-Bacterial Agents/pharmacology , Mastitis, Bovine/microbiology , Milk/microbiology , Staphylococcal Infections/veterinary , Staphylococcus aureus/drug effects , Animals , Argentina , Cattle , Drug Resistance, Microbial , Female , Mastitis, Bovine/drug therapy , Microbial Sensitivity Tests/veterinary , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcus aureus/isolation & purification
14.
Eur J Biochem ; 255(1): 262-70, 1998 Jul 01.
Article in English | MEDLINE | ID: mdl-9692927

ABSTRACT

The conflicting data on the binding of the two molecules of ATP that are involved in the overall reaction catalyzed by carbamoyl-phosphate synthetase (CPS) of Escherichia coli, and a mechanism recently proposed for this reaction, has led us to reexamine ATP binding using pulse/chase techniques. With [gamma-32P]ATP and bicarbonate in the pulse solution, there is a positive intercept at zero time of approximately 1 mol Pi/mol CPS in the plot of 32Pi formation against time, irrespective of whether the incubation is terminated by the addition of acid or by addition of a chase solution containing glutamine, excess unlabeled ATP and bicarbonate. The intercept is decreased to about 50% if the excess unlabeled ATP is added prior to the addition of the glutamine. These are the expected results if the intercept reflects the reversible formation of enzyme-bound ADP and carboxyphosphate. Approximately 0.6 mol carbamoyl [32P]phosphate/mol enzyme is formed in these experiments when the pulse step is terminated by addition to the chase solution. The ATP molecule that provides the phosphoryl group of carbamoyl phosphate, therefore, also binds to the enzyme in the absence of ammonia or glutamine and reacts in the chase to give carbamoyl phosphate before it can dissociate from the enzyme. At 1 mM ATP, the binding of both ATP molecules is essentially complete at 2.5 s, but the dissociation of the ATP that yields carbamoyl phosphate is extremely slow (t(1/2) of about 6 min at 22 degrees C; HCO3-, 40 mM), although it is faster in the absence of bicarbonate. The extreme sequestration from the aqueous environment of this ATP allows the enzyme-ATP complex to be separated from the surrounding ATP by centrifugal gel filtration. After two successive steps of gel filtration through Sephadex G-50 equilibrated with unlabeled ATP and bicarbonate, the majority of the radioactivity remaining in the solution is bound to the enzyme and is released as [gamma-32P]ATP if acid is added, or is converted to carbamoyl [32P]phosphate by addition to chase solution, without concomitant release of 32Pi. K+ is necessary in the pulse solution, but not in the chase solution, to demonstrate this binding. These findings and other confirmatory experiments demonstrate conclusively that, in the presence of K+, both ATP molecules bind to the enzyme in the absence of ammonia or glutamine. The bound ATP that yields Pi in the overall reaction is replaced relatively rapidly by exchange and by hydrolysis in the bicarbonate-dependent ATPase activity of the enzyme, whereas the bound ATP that provides the phosphoryl group of carbamoyl phosphate is replaced very slowly. The temporal pattern of carbamoyl [32P]phosphate formation from [gamma-32P]ATP, in pulse/chase experiments in which a small concentration of ammonia is added to the pulse solution, shows that, in the normal enzyme reaction, this last ATP molecule binds to the enzyme before ammonia. These findings exclude a recently proposed mechanism [Kothe, M., Eroglu, B., Mazza, H., Samudera, H. & Powers-Lee, S. (1997) Proc. Natl Acad. Sci. USA 94, 12348-12353] in which a single molecule of ATP bound at the catalytic center phosphorylates bicarbonate and provides the phosphoryl group of carbamoyl phosphate. A mechanism in which a single ATP molecule binds, followed by the binding of bicarbonate and ammonia (from glutamine) and the release of Pi before the second molecule of ATP is bound is also excluded. We have previously reported very similar findings for carbamoyl-phosphate synthetase (ammonia), strongly suggesting that the different types of CPS share a common mechanism. The virtual sequestration of the ATP that provides the phosphoryl group of carbamoyl phosphate is consistent with a palmate-binding site, with the nucleotide bound within a beta-sheet sandwich, and a loop closure mechanism triggered by the binding of bicarbonate or the formation of carboxyphosphate.


Subject(s)
Adenosine Triphosphate/metabolism , Carbamoyl-Phosphate Synthase (Ammonia)/metabolism , Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)/metabolism , Carbamyl Phosphate/metabolism , Escherichia coli/enzymology , Ammonia/metabolism , Bicarbonates/metabolism , Cations, Monovalent/metabolism , Glutamine/metabolism , Models, Chemical , Ornithine/metabolism , Phosphates/metabolism , Potassium/metabolism
15.
Appl Environ Microbiol ; 63(3): 1165-7, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9055432

ABSTRACT

The killer properties of yeasts isolated from olive brines were examined in the absence and presence of sodium chloride in concentrations of up to 6% (wt/vol). An apparent enhancement of the killing action as the salt concentration increased, as well as changes in the spectra of activity against selected target strains, was observed in a few strains. Culture filtrates from killer strains grown at different NaCl concentrations (0, 3, or 6% [wt/vol]) were tested against sensitive yeasts cultivated under the same conditions. While the sensitivity of the target strain greatly increased in the presence of salt, no significant effect on toxin production was noticed.


Subject(s)
Sodium Chloride/pharmacology , Yeasts/drug effects , Dose-Response Relationship, Drug , Phenotype
16.
Int J Biol Macromol ; 18(4): 255-62, 1996 Jun.
Article in English | MEDLINE | ID: mdl-8739129

ABSTRACT

Hypoxanthine-guanine phosphoribosyltransferase (HGPRT, E.C.2.4.2.8) from Artemia cysts exhibits maximum activity at 70 degrees C. Its thermal stability has been examined following enzymatic activity as a function of temperature. Cold-induced renaturation experiments of samples heated at increasing temperatures showed that reversibility of thermal inactivation depends on the incubation time and final temperature. Prolonged incubation of the thermoinactivated enzyme at 0 degree C did not afford any further increase of the catalytic activity at 37 degrees C. The complex substrate PRPP:Mg protects HGPRT from thermal inactivation. However, incubations with hypoxanthine rendered a less thermostable enzyme at any temperature tested. The irreversible inactivation of HGPRT proceeds in two exponential steps. The analysis of the apparent rate constants for the fast and the slow phases, lambda 1 and lambda 2 as per the Lumry and Eyring model suggests the existence of more than three states in the thermal denaturation pathway of the free enzyme. In the presence of PRPP:Mg the irreversible process follows a single exponential and proceeds very slowly below 70 degrees C. PRPP:Mg also protects the enzyme from inactivation by NEM and pCMB, suggesting that -SH groups may be in the vicinity of the active site.


Subject(s)
Artemia/enzymology , Hypoxanthine Phosphoribosyltransferase/antagonists & inhibitors , Hypoxanthine Phosphoribosyltransferase/metabolism , Animals , Binding Sites , Enzyme Stability , Hypoxanthine Phosphoribosyltransferase/chemistry , Kinetics , Macromolecular Substances , Phosphoribosyl Pyrophosphate , Protein Denaturation , Protein Folding , Substrate Specificity , Temperature
17.
Rev Esp Cardiol ; 49(4): 281-7, 1996 Apr.
Article in Spanish | MEDLINE | ID: mdl-8650405

ABSTRACT

OBJECTIVE: This study was designed to investigate urate production by swine hearts using an in vivo regionally ischemic-reperfused model. ANIMALS AND METHODS: Ten female pigs underwent 60 minutes of myocardial ischemia by clamping of the left anterior descending artery and afterwards 120 minutes of reperfusion. Epicardial biopsies and blood samples from coronary sinus were taken before ligation, at the end of ischemic period and 5, 30, 60 and 120 minutes upon reperfusion. RESULTS: During ischemia, tissue levels of ATP and ADP greatly declined with a subsequent increase in the concentration of AMP, inosine and hypoxanthine (33 +/- 12 vs 93 +/- 17, 26 +/- 8 vs 768 +/- 86 and 32 +/- 10 vs 219 +/- 26 nmol/g dry weight, p < 0.01 for each). Despite the great increase in the hypoxanthine levels, uric acid concentration remained constant (69 +/- 9 vs 32 +/- 12 nmol/g dry weight, NS). Hypoxanthine, xanthine and uric acid concentrations increased in blood samples obtained from the coronary sinus at the end of ischemic period (17.99 vs 31.03 nmol/ml, p < 0.01, 0.29 vs 1.45 nmol/ml, p < 0.05 and 1.20 vs 2.31 nmol/ml, p < 0.01 respectively) and were enhanced upon reperfusion (35.8 and 3.89 nmol/ml for hypoxanthine and uric acid respectively, p < 0.05) without any significant modifications in their concentrations at the arterial level. CONCLUSION: These results demonstrate that the ischemic-reperfused swine heart produces urate probably outside the myocardium.


Subject(s)
Myocardial Reperfusion Injury/metabolism , Uric Acid/metabolism , Animals , Female , Free Radicals , Hypoxanthine , Hypoxanthines/blood , Myocardial Reperfusion Injury/blood , Myocardium/metabolism , Swine , Uric Acid/blood , Xanthine , Xanthines/blood
18.
Biochim Biophys Acta ; 1156(2): 128-34, 1993 Feb 13.
Article in English | MEDLINE | ID: mdl-8427871

ABSTRACT

Incorporation of the radiolabelled purine bases adenine, guanine and hypoxanthine into acid soluble fraction, RNA and DNA nucleotides during the early larval development of Artemia sp. was studied. Adenine was the best precursor and guanine the poorest. The adenine phosphoribosyltransferase (APRT) activity was considerably higher than that of hypoxanthine-guanine phosphoribosyltransferase (HGPRT) and these activities did not significantly change throughout larval development. The pattern of purine interconversion was dependent on naupliar age. Conversion of [14C]adenine and [14C]hypoxanthine into guanine nucleotides increased with time of development. However, the conversion of [14C]guanine into [14C]adenine nucleotides was very low.


Subject(s)
Artemia/metabolism , Purines/metabolism , Adenine/metabolism , Adenine Nucleotides/metabolism , Adenine Phosphoribosyltransferase/metabolism , Animals , Carbon Radioisotopes , Guanine Nucleotides/metabolism , Hypoxanthine , Hypoxanthine Phosphoribosyltransferase/metabolism , Hypoxanthines/metabolism , Larva/metabolism
19.
J Comp Physiol B ; 162(3): 263-6, 1992.
Article in English | MEDLINE | ID: mdl-1613165

ABSTRACT

In vivo studies of the incorporation of [U-14C]glycine into purine nucleotides have established the de novo pathway for purine biosynthesis in Artemia sp. during the early period of larval development. This pathway can be modified by the salt concentration of the incubation media. In addition, Artemia of different geographical origins may differ with respect to the detection, functionality and variability of this metabolical pathway.


Subject(s)
Artemia/metabolism , Purines/metabolism , Animals , Artemia/growth & development , Environment , Glycine/metabolism , Larva/metabolism , Purine Nucleotides/biosynthesis , Sodium Chloride
20.
Transpl Int ; 5 Suppl 1: S196-7, 1992.
Article in English | MEDLINE | ID: mdl-14621774

ABSTRACT

The initial enthusiasm for orthotopic liver transplantation (OLT) in patients with hepatocellular carcinoma (HCC) soon vanished as early recurrences appeared. OLT in HCC remains a controversial issue. We evaluated the efficacy of preoperative studies to select No-Mo patients and determined whether pT stage and histopathological grade (G) have a prognostic significance. A group of 25 patients, all previously thoroughly studied to rule out extrahepatic disease, underwent OLT for HCC. All patients were pNo after pathological study and none of the six patients who died in the postoperative period showed extrahepatic dissemination at necropsy (pMo). The recurrence rate was 43%. The 2 and 5 years actuarial survival was 62% and 43% respectively. The pT and G were not prognostic factors for long-term survival. We think that HCC is still a good indication for OLT because almost 50% of patients have good survival prospects.


Subject(s)
Carcinoma, Hepatocellular/surgery , Liver Neoplasms/surgery , Liver Transplantation/physiology , Carcinoma, Hepatocellular/pathology , Female , Follow-Up Studies , Humans , Liver Neoplasms/pathology , Liver Transplantation/mortality , Male , Neoplasm Staging , Prognosis , Retrospective Studies , Survival Analysis , Time Factors
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