PhTX-II a basic myotoxic phospholipase A2 from Porthidium Hyoprora snake venom, pharmacological characterization and amino acid sequence by mass spectrometry

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOA monomeric basic PLA2 (PhTX-II) of 14149.08 Da molecular weight was purified to homogeneity from Porthidium hyoprora venom. Amino acid sequence by in tandem mass spectrometry revealed that PhTX-II belongs to Asp49 PLA2 enzyme class and displays conserved domains as the catalytic network, Ca2+-binding loop and the hydrophobic channel of access to the catalytic site, reflected in the high catalytic activity displayed by the enzyme. Moreover, PhTX-II PLA2 showed an allosteric behavior and its enzymatic activity was dependent on Ca2+. Examination of PhTX-II PLA2 by CD spectroscopy indicated a high content of alpha-helical structures, similar to the known structure of secreted phospholipase IIA group suggesting a similar folding. PhTX-II PLA2 causes neuromuscular blockade in avian neuromuscular preparations with a significant direct action on skeletal muscle function, as well as, induced local edema and myotoxicity, in mice. The treatment of PhTX-II by BPB resulted in complete loss of their catalytic activity that was accompanied by loss of their edematogenic effect. On the other hand, enzymatic activity of PhTX-II contributes to this neuromuscular blockade and local myotoxicity is dependent not only on enzymatic activity. These results show that PhTX-II is a myotoxic Asp49 PLA2 that contributes with toxic actions caused by P. hyoprora venom.A monomeric basic PLA2 (PhTX-II) of 14149.08 da molecular weight was purified to homogeneity from Porthidium hyoprora venom. Amino acid sequence by in tandem mass spectrometry revealed that PhTX-II belongs to Asp49 PLA2 enzyme class and displays conserved domains as the catalytic network, Ca2+-binding loop and the hydrophobic channel of access to the catalytic site, reflected in the high catalytic activity displayed by the enzyme. Moreover, PhTX-II PLA2 showed an allosteric behavior and its enzymatic activity was dependent on Ca2+. Examination of PhTX-II PLA2 by CD spectroscopy indicated a high content of alpha-helical structures, similar to the known structure of secreted phospholipase IIA group suggesting a similar folding. PhTX-II PLA2 causes neuromuscular blockade in avian neuromuscular preparations with a significant direct action on skeletal muscle function, as well as, induced local edema and myotoxicity, in mice. The treatment of PhTX-II by BPB resulted in complete loss of their catalytic activity that was accompanied by loss of their edematogenic effect. On the other hand, enzymatic activity of PhTX-II contributes to this neuromuscular blockade and local myotoxicity is dependent not only on enzymatic activity. These results show that PhTX-II is a myotoxic Asp49 PLA2 that contributes with toxic actions caused by P. hyoprora venom630773097FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2009/51207-

Caracterización bioquímica del veneno de la serpiente Bothrops roedingeri Mertens, 1942, y sus actividades edematógena, hemorrágica y miotóxica

Introduction: Snakebite envenoming is considered by the World Health Organization (WHO) as a neglected tropical disease. Currently, Bothrops snake venoms are being studied intensively, but there is little knowledge about Bothrops roedingeri venom.Objectives: To biochemically characterize B. roedingeri total venom and evaluate its myotoxic, edematogenic, and hemorrhagic activity.Materials and methods: We characterized B. roedingeri venom enzymatic activity by determining the phospholipase A2 and the proteolytic and fibrinogenolytic action using SDSPAGE electrophoresis while we characterized its venom toxicity by determining the minimum hemorrhagic dose, the minimum edema dose, and the local and systemic myotoxic effects.Results: Bothrops roedingeri venom showed a PLA2 activity of 3.45 ± 0.11 nmoles/min, proteolytic activity of 0.145 ± 0.009 nmoles/min, and a fibrinogen coagulation index of 6.67 ± 1.33 seconds. On the other hand, it produced an minimum hemorrhagic dose of 24.5 μg, an minimum edema dose of 15.6 μg, and a pronounced local myotoxic effect evidenced by the elevation of plasma creatine kinase levels after intramuscular inoculation. The venom showed no systemic myotoxicity.Conclusions: Bothrops roedingeri venom has local hemorrhagic, edematogenic, and myotoxic activity. Enzymatically, it has high PLA2 activity, which would be responsible for the myotoxic and edematogenic effects. It also has proteolytic activity, which could affect coagulation given its ability to degrade fibrinogen, and it causes bleeding through the metalloproteases.Introducción. El envenenamiento por mordedura de serpiente es considerado por la Organización Mundial de la Salud (OMS) una enfermedad tropical desatendida. Si bien los venenos de otras serpientes Bothrops se vienen estudiado ampliamente, poco se conoce del de Bothrops roedingeri.Objetivos. Caracterizar bioquímicamente el veneno total de la serpiente B. roedingeri y evaluar su actividad miotóxica, edematógena y hemorrágica.Materiales y métodos. Se hizo la caracterización enzimática del veneno de B. roedingeri determinando la actividad de la fosfolipasa A2 (PLA2) y de las enzimas proteolíticas, así como su acción fibrinogenolítica mediante electroforesis en gel de poliacrilamida con duodecilsulfato sódico (sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDSPAGE), y la caracterización tóxica del veneno estableciendo la dosis hemorrágica mínima, la dosis edematógena mínima y el efecto miotóxico local y sistémico.Resultados. La actividad de las PLA2 del veneno total de B. roedingeri fue de 3,45 ± 0,11 nmoles/minuto, la proteolítica, de 0,145 ± 0,009 nmoles/minuto, en tanto que el índice de coagulación del fibrinógeno fue de 6,67 ± 1,33 segundos. Por otro lado, el veneno produjo una dosis hemorrágica mínima de 24,5 μg, una dosis edematógena mínima de 15,6 μg y un pronunciado efecto miotóxico local evidenciado por la elevación de los niveles plasmáticos de creatina cinasa después de la inoculación por vía intramuscular. No se registró miotoxicidad sistémica.Conclusiones. El veneno de B. roedingeri tiene efectos hemorrágicos, edematógenos y miotóxicos locales, así como una elevada actividad de la PLA2, que sería responsable de los efectos miotóxico y edematógeno. También presentó actividad proteolítica, la cual podría afectar la coagulación, dada su capacidad para degradar el fibrinógeno y producir hemorragia por acción de las metaloproteasas

Psychometric evaluation and invariance of the Spanish version of the Block Fat Screener (BFS-E) in university students

IntroductionUniversity students constantly face a number of health challenges related to an unhealthy diet, characterized by a high intake of saturated fats.ObjectiveThis study aimed to analyze the psychometric properties of the Spanish version of the Block Fat Screener (BFS-E) food frequency questionnaire in a university population.MethodsAn observational analytical study of instrumental type was carried out in 5608 Peruvian university students. Based on the Block Fat Screener questionnaire, a back-translation and cultural adaptation process was carried out. The validity of the questionnaire was determined through exploratory factor analysis (EFA) and confirmatory factor analysis (CFA), hypothesizing a unidimensional structure. For the determination of reliability, the alpha coefficients were considered; likewise, the ω and H coefficients were used to evaluate the construct. The model explained 63% of the cumulative variance.ResultsThe CFA confirmed the unidimensionality of the 16-item questionnaire with appropriate goodness-of-fit indicators; therefore, which model of the Peruvian version adequately fits the observed data. The values of the reliability coefficients were higher than 0.90, with ordinal α = 0.94, ω = 0.94, and H = 0.95.ConclusionThe Spanish version of the Block Fat Screener food frequency questionnaire presents adequate psychometric properties and is therefore a valid scale to quickly measure fat intake in university students in a Latin American context

ACP-TX-I and ACP-TX-II, Two Novel Phospholipases A2 Isolated from Trans-Pecos Copperhead Agkistrodon contortrix pictigaster Venom: Biochemical and Functional Characterization

This work reports the purification and biochemical and functional characterization of ACP-TX-I and ACP-TX-II, two phospholipases A2 (PLA2) from Agkistrodon contortrix pictigaster venom. Both PLA2s were highly purified by a single chromatographic step on a C18 reverse phase HPLC column. Various peptide sequences from these two toxins showed similarity to those of other PLA2 toxins from viperid snake venoms. ACP-TX-I belongs to the catalytically inactive K49 PLA2 class, while ACP-TX-II is a D49 PLA2, and is enzymatically active. ACP-TX-I PLA2 is monomeric, which results in markedly diminished myotoxic and inflammatory activities when compared with dimeric K49 PLA2s, confirming the hypothesis that dimeric structure contributes heavily to the profound myotoxicity of the most active viperid K49 PLA2s. ACP-TX-II exhibits the main pharmacological actions reported for this protein family, including in vivo local myotoxicity, edema-forming activity, and in vitro cytotoxicity. ACP-TX-I PLA2 is cytotoxic to A549 lung carcinoma cells, indicating that cytotoxicity to these tumor cells does not require enzymatic activity

'PLA2 D49 and K49 mitoxins from the venom of Bothrops brazili snake : purification and structural and funcional characterization

Orientadores: Sergio Marangoni, Luis Alberto Ponce SotoDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: As enzimas PLA2 provenientes de veneno de serpentes são intensamente estudadas devido a que os envenenamentos constituem um dos principais problemas de saúde em muitos paises. Por outro lado, estas toxinas ajudam a revelar aspectos desconhecidos da fisiologia celular e tisular. Neste trabalho, apresentamos a purificação e a caracterização bioquímica e farmacológica de duas miotoxinas fosfolipases A2: BbTX-II e BbTX-III, a partir de veneno de Bothrops brazili. As duas proteínas foram isoladas e purificadas usando um procedimento simples e rápido envolvendo duas etapas cromatográficas, exclusão molecular em Sephadex G-75 e HPLC de fase reversa (C18). A eletroforese de ambas miotoxinas mostrou massas relativas em torno de 13 e 27 kDa (para monômeros e dímeros respectivamente). Espectrometria de massa por MALDI-TOf confirmou a pureza das proteínas e mostrou que possuem massas moleculares em torno de 13,8 kDa. A análise de aminoácidos mostrou alto conteúdo de aminoácidos básicos e hidrofóbicos, assim como 14 resíduos de Cys. BbTX-III apresentou atividade PLA2 na presença de um substrato cromogênico, mostrando comportamento sigmoidal, principalmente à baixas concentrações. Atividade máxima foi alcançada em pH 8 e entre 35-45 oC. BbTX-III mostrou-se completamente dependente de Ca2+ e, na presença dos íons Mg2+, Mn2+, Cd2+ e Zn2+, a atividade enzimática foi reduzida a níveis similares aos observados na ausência de Ca2+. A análise de composição de aminoácidos mostrou alta presença de Lys, His e Arg (pI 8,46). A presença de 14 resíduos de cisteína sugere a formação de 7 pontes dissulfeto. O estudo de homologia da seqüência da PLA2 BbTX-III mostrou que existem posições extremamente conservadas nas PLA2. S(1), L(2), E(4) 7 a 10 (QMIL), Y(21). Os resíduos conservados Y(28), G(30), G(32), D(49), H(48) e Y(52) estão direta ou indiretamente ligados com a catálise. Além disso, BbTX-III apresentou algumas mutações: K(35) -> G(35), R(51) -> Y(51) e D(118) -> A(118) que estão estrategicamente posicionadas para a expressão da atividade catalítica. Apesar destas substituições, as atividades farmacológicas e a atividade catalítica são mantidas. O efeito neurotóxico de BbTX-III foi analisado in vitro na preparação neuromuscular biventer cervicis de pintainhos. O resultado mostrou que a toxina é menos potente quando comparada com venenos crotálicos. BbTX-III demonstrou efeito miotóxico local in vivo através da liberação de creatina quinase (CK) e, efeito inflamatório, através de edema de pata. Como a BbTX-III produziu forte efeito inflamatório, a hidrólise de fosfolipídios poderia ser relevante neste fenômeno. BbTX-II foi caracterizada como uma PLA2 K49 (cataliticamente inativa) em função das características físico-químicas evidenciadas: massa de 13,68 kDa, 121 resíduos de aminoácidos, caráter básico (pI 8.73) e alto grau de homologia seqüencial na sua estrutura primária, quando comparada com outras PLA2B K49 procedentes de veneno de serpentes botrópicas. O alinhamento com outras seqüências completas de PLA2 BK49 mostrou a presença de algumas mutações importantes. Assim, as substituições Y?N(27), N?P(58) e L?F(114) não modificaram os efeitos biológicos aqui estudados, revelando que poderiam estar relacionados com outras atividades. Os resíduos N(28), K(111), L(32) poderiam contribuir com a interrupção da catálise. Esta nova PLA2 K49 BbTX-II mostrou miotoxicidade local in vivo, atividade inflamatória e letalidade, corroborando que se enquadra dentro da família de proteínas PLAB2B K49. Os estudos de neurotoxicidade revelaram um efeito neurotóxico in vitro na preparação biventer cervicis de pintainho (20 µg/ml). BbTX-II e BbTX-III mostraram ser miotoxinas com atividade edematogênica e neurotóxica, independentemente de apresentarem atividade catalítica (BbTX-III) ou não (BbTX-II) Apoiando a existência de regiões moleculares distintas à catalítica responsáveis pelos efeitos farmacológicos. Os efeitos farmacológicos da toxina BbTX-III (PLA2B D49) provavelmente tenham uma estrita relação entre a atividade enzimática e a ligação da toxina com micro-domínios na membrana plasmática onde sua atividade seja maximizada e cause danos relevantes na organização da membrana. No caso da BbTX-II (PLAB2 K49) possivelmente a combinação de aminoácidos aromáticos/hidrofóbicos e positivamente carregados da região C-terminal seja a responsável de alterar a integridade da membrana plasmática.Abstract: The enzymes PLA2 coming of venom snake are studied intensely due to that the poisonings constitute one of the main problems of health in many countries. On the other hand, these toxins help to reveal unknown aspects of the cellular and tissue physiology. In this work, we presented the purification and biochemical and pharmacological characterization of two myotoxic phospholipases A2: BbTX-II and BbTX-III from Bothrops brazili venom. The two proteins were isolated and purified using a simple and fast procedure involving two chromatographic steps, molecular exclusion in Sephadex G-75 and reverse-phase HPLC (C-18 column). Both myotoxins showed around 13 and 27 kDa (for monomers and dimers, respectively) relative mass. MALDI-TOf mass spectrometry confirmed the purity of the proteins showing molecular masses around 13,8 kDa. Amino acid analysis showed a high content of hydrophobic and basic amino acids as well as 14 cysteine residues. BbTX-III presented PLA2 activity in the presence of a chromogenic substrate, showing sigmoidal behavior, mainly at low concentrations. Maximum PLA2 activity was reached at pH 8 and 35-45 oC. Maximum activity required Ca2+ and, in the presence of Mg2+, Mn2+, Cd2+ and Zn2+, was reduced at similar levels as observed in the absence of Ca2+. Amino acids analysis of composition showed high presence of Lys, His and Arg (pI 8,46). The presence of 14 cystein residues suggested the formation of 7 disulfide bridges. Sequence homology of the PLA2 BbTX-III revealed positions extremely conserved in PLA2 S(1), L(2), E(4) 7 to 10 (QMIL), Y(21). The conserved residues Y(28), G(30), G(32), D(49), H(48) and Y(52) are direct or indirectly linked to the catalysis. Besides, BbTX-III presented some mutations: K(35) -> G(35), R(51) -> Y(51) and D(118) -> A(118) that are strategically positioned for the expression of catalytic activity. Despite these substitutions, the pharmacological and catalytic activities are maintained. The neurotoxic effect of BbTX-III was analyzed in vitro at chick biventer cervicis muscle preparation. Our results showed that the blockage of the muscle contraction was lower when compared with crotalic venoms. BbTX-III demonstrated in vivo myotoxic local effect through the liberation of creatine kinase (CK) and inflammatory effect through paw edema. As BbTX-III produced strong inflammatory effect, the phospholipids hydrolysis could be relevant in this phenomenon. BbTX-II was characterized as PLA2 homologous K49 (catalytically inactive), because its chemical and physical evidenced characteristics: mass of 13,68 kDa, 121 amino acids residues, basic character (pI 8.73) and high sequential homology in its primary structure, when compared with other PLA2 K49 from venom of Botrhops serpents. The alignment with other complete sequences of PLA2 homologous K49 showed the presence of some important mutations. Substitutions Y->N(27), N->P(58), and L->F(114) did not modify the biological activities here studied, revealing that it could be related to other activities. The residues N(28), K(111), L(32) could contribute with the interruption of the catalysis. This new PLA2 K49 BbTX-II showed in vivo myotoxic local effect, inflammatory and lethality activities, evidencing it was fitted to the family of proteins PLA2 K49 homologous. Beside BbTX-II revealed in vitro neurotoxyc effect at chick biventer cervicis muscle preparation (20 µg/mL). BbTX-II and BbTX-III showed to be myotoxins to activity edematogenic and neurotoxyc independently of present catalytic activity (BbTX-III) or no (BbTX-II) Supporting the existence of molecular areas different to the catalytic responsible for the pharmacological effects. The pharmacological effects of the toxin BbTX-III (PLA2 D49) they probably have a strict relationship between the enzymatic activity and binding of the toxin with micro-domains in the plasmatic membrane where the activity is maximized and cause relevant damages in the organization of the membrane. In the case of the BbTX-II (PLA2 K49) possibly the combination of amino acids aromatics/hidrophobics and positively loaded of the area C-terminal it is the responsible of altering the integrity of the plasmatic membrane.MestradoBioquimicaMestre em Biologia Funcional e Molecula

Chemical modifications and pharmacological characterization of PhTX-I and PhTX-II myotoxins isolated from Porthidium hyoprora

Orientadores: Sergio Marangoni, Luís Alberto Ponce SotoTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: No presente trabalho, a partir do veneno de Porthidium hyoprora, foram purificadas duas fosfolipases A2 (PLA2) miotóxicas, denominadas PhTX-I e PhTX-II, determinadas suas sequências primárias e caracterizadas bioquímica e farmacologicamente. Ambas as proteínas foram purificadas em uma única etapa cromatográfica (HPLC-FR), sendo obtida homogênea e com alto grau de pureza, confirmado por SDS-PAGE, análise de aminoácidos e espectrometria de massas. PhTX-I e PhTX-II estão constituídas de uma única cadeia polipeptídica e possuem massas moleculares de 14.249 e 14.149 Da, respectivamente. A sequência de aminoácidos foi determinada via espectrometria de massas ESI-MS/MS, pertencendo à família das PLA2 D49. A estrutura secundária predominante destas proteínas está constituída de ?-hélices. PhTX-I e PhTX-II induziram atividade paralisante neuromuscular ex vivo na preparação de biventer cervicis de pintainho e esta atividade foi similar ao efeito de outras PLA2 de veneno botrópico. In vivo, tanto PhTX-I quanto PhTX-II, induziram elevada miotoxicidade local e atividade edematogênica. Adicionalmente, PhTX-I induz liberação de IL-6 após injeção intramuscular em camundongos. In vitro, PhTX-I e PhTX-II foram citotóxicas de maneira dose-dependente para as linhagens celulares de fibroblastos NIH-3T3 e NG97 derivada de astrocitoma humano grau III. Adicionalmente PhTX-I foi citotóxica sobre cultura celular de miotubos C2C12, entretanto teve pouco efeito citolítico sobre mioblastos de músculo esquelético. Foram feitas modificações químicas em resíduos de aminoácidos específicos da PhTX-I e avaliadas suas atividades farmacológicas após as modificações. Foram modificados um resíduo de His e Trp, quatro de Tyr e sete de Lys. A análise destas proteínas por dicroísmo circular demonstrou que a estrutura secundária das proteínas modificadas não foi alterada significantemente. Avaliação das atividades biológicas indicam um papel crítico desempenhado pelos aminoácidos Lys e Tyr na miotoxicidade, efeito paralisante neuromuscular e principalmente na citotoxicidade induzida por PhTX-I. A atividade catalítica da PhTX-I é relevante para os efeitos edematogênico, paralisante neuromuscular e miotóxico, mas não para sua atividade citotóxica, pois este efeito foi completamente independente da atividade catalítica de PhTX-I, evidenciando a existência de regiões moleculares, diferentes a do sítio catalítico, as quais podem ser responsáveis por pelo menos algumas das propriedades farmacológicas da PhTX-I. Podemos concluir, que apesar de ser demonstrada uma dissociação parcial, tanto o sítio catalítico como os sítios farmacológicos hipotéticos são relevantes para o perfil farmacológico de PhTX-IAbstract: In this work, were purified two myotoxics phospholipases A2 (PLA2), PhTX-I and PhTX-II, from P. hyoprora snake venom, determined their primary sequences and characterized their pharmacological and biochemical activities. These proteins were purified in a single chromatographic step (FR-HPLC) and obtained with homogeneous and high purity, which was confirmed by SDS-PAGE, amino acid analysis and mass spectrometry. PhTX-I and PhTX-II are constituted of a single polipetide chain and have molecular mass of 14.249 and 14.149 Da, respectively. The amino acid sequence was determined by ESI-MS/MS mass spectrometry; belong to PLA2 D49 family. The predominant secondary structure of these proteins consists of ?-helix. PhTX-I and PhTX-II induced neuromuscular paralyzing effect ex vivo in young chick biventer cervicis preparations, this activity was similar to other PLA2 of snake venom. In vivo, PhTX-I and PhTX -II induced local myotoxicity and edema-forming activity, in addition, PhTX-I induces release of IL-6 after intramuscular injection in mice. In vitro, PhTX-I and PhTX -II were cytotoxic to the fibroblast cell line NIH-3T3 and NG97 derived from grade III human astrocytoma. Additionally PhTX-I induced low cytotoxicity in skeletal muscle myoblasts, however was able to lyse myotubes. Were made modifications chemical in PhTX-I specific amino acid residues and evaluated the pharmacological activities after modifications. Was modified one His and Trp, four Tyr and seven Lys residues? Analysis by circular dichroism demonstrated that secondary structure of protein remains practically unchanged. Evaluation of biological activities indicate a critical role of Lys and Tyr amino acid at myotoxicity, neuromuscular paralyzing effect, and especially cytotoxicity induced by PhTX-I. The catalytic activity of PhTX-I is relevant to the edematogenic, neuromuscular paralyzing and myotoxic effects, but not for their cytotoxic activity, since this effect is completely independent of the catalytic activity of PhTX-I, demonstrating the existence of molecular regions, different from the catalytic site, which may be responsible for at least some of the pharmacological properties of PhTX-I. We conclude that both the catalytic site and the hypothetical pharmacological site(s) are apparently relevant for the pharmacological profile of PhTX-I, although a partial dissociation between these activities has been demonstratedDoutoradoBioquimicaDoutor em Biologia Funcional e Molecula

Association Between Perceived Access to Healthcare and the Perception of Illness Among Peruvian Adults with Chronic Diseases During COVID-19 Pandemic

The COVID-19 pandemic has greatly affected the provision of care for patients with chronic diseases. Due to social restrictions and reductions in contact with health services, the negative perception of chronic disease is expected to have increased. The aim of this study was to determine the association between perceived access to healthcare and the perception of illness among Peruvian population with chronic disease. It was a cross-sectional analytical study, with a sample of 987 inhabitants to whom the questionnaires “Coverage of health services” and “The Brief Illness Perception Questionnaire” (BIPQ) were applied. Having health insurance (PRa = 0.683; 95% CI = 0.613-0.761) acts as a protective factor for a positive illness perception of chronic disease, however, a waiting time greater than 3 months to obtain a medical appointment (PRa = 1.417; 95% CI = 1.319-1.522) and poor access to health services (PRa = 1.435; 95% CI = 1.226-1.681) resulted in the probability of a negative illness perception of chronic disease. Thus, there is an association between perceived poor access to healthcare and the negative illness perception of chronic disease in Peruvian population during pandemic COVID-19

Comparación de los perfiles de resistencia antimicrobiana de Escherichia coli uropatógena e incidencia de la producción de betalactamasas de espectro extendido en tres establecimientos privados de salud de Perú

Introduction: The appearance of multidrug-resistant and beta-lactamase producing enterobacteria in outpatient care facilities represent a public health problem in Perú.Objective: To compare the resistance profiles of uropathogenic Escherichia coli and to identify extended-spectrum beta-lactamase-producing phenotypes in three private health facilities located in the Peruvian coast, Andean and jungle regions.Materials and methods: We conducted a descriptive study on 98 urine samples from Lima (coast), Juliaca (Andean region) and Iquitos (jungle region) during 2016. We determined the antimicrobial susceptibility in 35 samples from Lima, 38 from Juliaca and 25 from Iquitos using eight antibiotic disks in samples from patients diagnosed with urinary infection. We also evaluated the production of extended-spectrum beta-lactamases with cefotaxime and ceftazidime disks and a combination of both with clavulanic acid on Mueller-Hinton agar.Results: We identified 18 resistance profiles ranging from those sensitive to others simultaneously resistant to seven antibiotics: 18.4% resistant to one and 54.0% to multiple antibiotics. We detected beta-lactamase production in 28.6% of the strains from the Puno region. Likewise, we observed a greater number of cases with resistance to ceftazidime, ceftriaxone, gentamicin, and trimethoprim-sulfamethoxazole in Puno’s health facility in patients within the 31 to 45 year age range.Conclusion: Resistance profiles varied according to the geographical location of the health facilities under study. Resistance to antibiotics was higher in the Andean region with 28.6% of strains producing extended-spectrum beta-lactamases.Introducción. La aparición de enterobacterias multirresistentes y productoras de betalactamasas de espectro extendido en pacientes ambulatorios con infecciones urinarias representa un problema de salud pública en Perú.Objetivo. Comparar los perfiles de resistencia de Escherichia coli uropatógenas e identificar los fenotipos de cepas productoras de betalactamasas de espectro extendido en tres establecimientos privados de salud localizados en las regiones de la costa, la sierra y la selva de Perú.Materiales y métodos. Se llevó a cabo durante el 2016 un estudio descriptivo de 98 muestras de orina de pacientes con infección urinaria, 35 procedentes de Lima (costa), 38 de Juliaca (sierra) y 25 de Iquitos (selva), en el que se determinó la sensibilidad antimicrobiana utilizando ocho discos antibióticos.Asimismo, se evaluó la producción de betalactamasas de espectro extendido con discos de cefotaxima, de ceftazidima o de su combinación, con ácido clavulánico en agar Mueller-Hinton.Resultados. Se identificaron 18 perfiles de resistencia que incluían desde los sensibles a todos los antibióticos hasta los resistentes simultáneamente a siete antibióticos, con el 18,4 % de aislamientos resistentes a un antibiótico y el 54,0 % de multirresistentes. Se detectó producción de betalactamasas en el 28,6 % de las cepas procedentes de la región de Puno. También, se observó un mayor número de casos en el rango de edad de 31 a 45 años con resistencia a ceftazidima, ceftriaxona, gentamicina y trimetoprim-sulfametoxazol en el establecimiento de salud de Puno.Conclusión. Los perfiles de resistencia variaron según la localización geográfica del establecimiento de salud