Adenocarcinoma de vesícula biliar: un caso de presentación temprana / Gallbladder adenocarcinoma: an early-stage case

El cáncer de la vesícula biliar es una enfermedad rara, con una incidencia mundial de 2 casos por cada 100 000 individuos con un pronóstico desfavorable. Con el aumento de colecistectomías por causas benignas, se ha incrementado la detección incidental de neoplasias vesiculares en las piezas quirúrgicas, siendo este el método diagnóstico más frecuente, generando retrasos en el manejo y requiriendo reintervenciones extensas. Debido a lo anterior, se resalta la importancia de un diagnóstico temprano preoperatorio, con el objetivo de ofrecer un tratamiento quirúrgico potencialmente curativo. Se presenta el caso de un paciente masculino de 72 años con un cuadro intermitente de dolor abdominal y pérdida de peso de un año de evolución, el cual fue diagnosticado con cáncer vesicular en etapa temprana y sometido a una colecistectomía laparoscópica extendida con linfadenectomía y hepatectomía parcial con una evolución a 6 meses sin complicaciones y bajo un protocolo de vigilancia periódica.

Gallbladder cancer is a rare disease, accounting a global incidence of 2 cases per 100 000 individuals with an unfavorable prognosis. The rise in cholecystectomies for benign causes has increased an incidental detection of vesicular neoplasms in the surgical specimens, being the main diagnostic method, therefore it generated delay in the management, requiring extensive re-interventions. It is important to improve early preoperative diagnosis, with the aim of offering a potentially curative surgical treatment. We present a case of a 72-year-old male with intermittent abdominal pain and weight loss of one year of evolution, who was diagnosed with early stage gallbladder cancer and underwent an extended laparoscopic cholecystectomy with lymphadenectomy and partial hepatectomy with a 6 months evolution without complications and under a periodic surveillance protocol.

Unique long-term simultaneous complications of conventional Roux-en-Y gastric bypass after 27 years: A case report.

INTRODUCTION: We report a case of late concomitant complications caused by conventional Roux-en-Y gastric bypaas and its managements. PRESENTATION OF CASE: A 62-year-old male presented 27 years after conventional gastric bypass Y-de-Roux (BGYR) with, sudden, moderate intensity abdominal pain, nausea, biliary vomiting and hyporexia. Persistent abdominal pain was constant, so a thoracoabdominal tomography was requested by the surgeon. It confirmed the presence of intestinal intussusception associated with lithiasis and cholecystitis. The patient reported having lost 45 kg since the BGYR. He goes to the operating room for definitive management. DISCUSSION: The etiology of post-BGYR intussusception is largely unknown, and multiple hypotheses have been created, such as the iatrogenic stitch created by the suture line in the entero-enteric anastomosis and the most common pattern found is antegrade. The use of contrasted CT as the most reliable diagnostic method. CONCLUSION: The importance of taking into account the possible complications existing in bariatric patients and their frequency gives us the opportunity to suspect and detect them in time and in the most cases the management must be surgical.

Diagnosis and surgical intervention of acute appendicitis secondary to enterobius vermicularis: case report.

Acute appendicitis is the most common cause of acute abdominal pain in the pediatric population. The physiopathology is secondary to luminal obstruction of the appendix by several causes, among these, one of the rarest is due to parasitosis by Enterobius vermicularis in 0.005-3 %. We hereby present a case of a 10-year-old Latin male with abdominal pain with irradiation to right iliac fossa, nausea, vomiting and fever, following the national Mexican guides for ultrasound and blood tests were ordered, patient was treated with laparoscopic appendectomy with intraoperative finding of live helminths, which pathology examination revealed to be Enterovirus vermicularis. The patient has an uneventful postoperative course and was discharged after completing antiparasitic therapy. The lessons from this case are the importance of a good and quick diagnosis for acute abdomen in children, the possibility of a parasitic entity in the finding of eosinophilia with appendicitis and the good results with the correct surgical treatment.

Evaluación de la toxicidad de chalconas sintéticas con potencial anti-Leishmania en ratones BALB/c / Toxicity assessment of synthetic chalcones with antileishmanial potential in BALB/c mice

RESUMEN Objetivo: Evaluar la toxicidad de tres chalconas sintéticas administradas por vía intraperitoneal en ratones BALB/c. Materiales y métodos: La dosis letal media (DL50) se estimó por el método Up-and-Down de Dixon. La toxicidad subcrónica de las chalconas se evaluó a 20 y 40 mg/kg por 21 días. Se evaluó el efecto tóxico a nivel de comportamiento, fisiológico, bioquímico e histológico. Resultados: La chalcona 43 generó moco en las heces, daño visceral (hígado) y alteración en el coeficiente de órganos (riñón, p = 0,037 y cerebro, p = 0,008) en comparación con el grupo control. Además, en el análisis histológico se observó que esta chalcona produjo edema, inflamación y necrosis en los órganos evaluados, aunque no hubo diferencia significativa con el control. Todos los parámetros bioquímicos no difirieron significativamente entre los grupos de tratamiento a dosis de 40 mg/kg y el control. Conclusiones: La DL50 para las tres chalconas fue superior a 550 mg/kg de peso corporal. Las chalconas 40 y 42 son relativamente no tóxicas. Ambas pueden considerarse seguras para la aplicación vía intraperitoneal en ratones BALB/c y, en consecuencia, son posibles candidatas para ser usadas en el tratamiento contra las leishmaniosis.

ABSTRACT Objective: To evaluate the toxicity of three synthetic chalcones administered intraperitoneally to BALB/c mice. Materials and methods: The median lethal dose (LD50) was estimated by Dixon's Up-and-Down method. Subchronic toxicity of chalcones was evaluated at 20 and 40 mg/kg for 21 days. Behavioral, physiological, biochemical, and histological toxic effects were evaluated. Results: Chalcone 43 produced mucus in feces, visceral damage (liver) and alterations in organ coefficient (kidney, p = 0.037 and brain, p = 0.008) when compared to the control group. In addition, histological analysis showed that this chalcone produced edema, inflammation and necrosis in the evaluated organs, although there was no significant difference with the control. None of the biochemical parameters differed significantly between the treatment groups at 40 mg/kg dose and the control. Conclusions: The LD50 for all three chalcones was greater than 550 mg/kg of body weight. Chalcones 40 and 42 were found to be relatively non-toxic. Both can be considered safe for intraperitoneal application in BALB/c mice and, consequently, are potential candidates for use in the treatment of leishmaniasis.

[Disseminated histoplasmosis and haemophagocytic syndrome in HIV patients: A case series in a Peruvian hospital]. / Histoplasmosis diseminada y síndrome hemofagocítico en pacientes VIH: serie de casos en un hospital peruano.

BACKGROUND: Disseminated histoplasmosis (DH) is an opportunistic fungal infection in severely immunocompromised patients with HIV infection. Haemophagocytic syndrome (HFS), which can occur in these co-infected patients when the immune response is significantly altered, is often associated with high mortality. AIMS: To describe the epidemiological, clinical, analytical and microbiological characteristics, along with studying the presence of HFS, in patients with DH-HIV. METHODS: A retrospective study was conducted on a case series using data from the clinical records of patients diagnosed with DH and HIV infection during the years 2014 and 2015. RESULTS: DH was diagnosed in 8 (1.3%) of 597 HIV patients. All patients were in stage C3, and 75% (6/8) were not receiving combined antiretroviral therapy (CART). The remaining two patients had recently begun CART (possible immune reconstitution syndrome). Five (62.5%) of the 8 patients met criteria for HFS. The most frequent clinical symptoms were lymphoproliferative and consumptive syndrome, respiratory compromise, and cytopenia. Histoplasma was isolated in lymph nodes of 75% (6/8) of the patients, in blood samples in 25% (2/8), and also in intestinal tissue in one patient. The antifungal therapy was amphotericin B deoxycholate, without adjuvants. The overall mortality was 50%. CONCLUSIONS: In this case series, DH-HIV co-infection frequently progressed to HFS with high mortality. The clinical picture may resemble that of other systemic opportunistic infections, such as tuberculosis, or can take place simultaneously with other infections. Clinical suspicion is important in patients with severe cytopenia and lymphoproliferative and consumptive syndrome in order to establish an early diagnosis and prescribing a timely specific therapy.

Classification System for Cardiorespiratory Fitness Based on a Sample of the Brazilian Population

Peak oxygen consumption (VO2peak) is an important prognostic marker and its classification helps the cardiologist in the therapeutic decision-making process. The most commonly used cardiorespiratory fitness (CRF) classification has not been validated for the Brazilian population. Objective: To elaborate a CRF classification using a Brazilian sample and to compare it with the American Heart Association (AHA), Cooper and UNIFESP classifications. Methods: A total of 6,568 healthy subjects were analyzed through cardiopulmonary exercise testing (CPET). They were distributed by sex and the following age groups (years): 7-12, 13-19, 20-79 (per decades) and > 80 years. After measurement of the VO2peak, participants were distributed into quintiles of CRF in very poor, poor, moderate, high and very high (AEMA Table). The CRF classifications by AEMA, AHA, Cooper, and UNIFESP were compared using the Wilcoxon, Kappa and concordance percentages. Results: VO2peak presented an inverse and moderate correlation with age considering both sexes (R = -0.488, p < 0.001). All paired comparisons between CRF classification systems showed differences (p < 0.001) and disagreement percentage - AEMA versus AHA (k = 0.291, 56.7%), AEMA versus Cooper (k = 0.220, 62.4%) and AEMA versus UNIFESP (k = 0.201, 63.9 %). Conclusion: The AEMA table showed important discrepancies in the classification of CRF when compared to other tables widely used in our setting. Because it was obtained from a large sample of the Brazilian population, the AEMA table should be preferred over other classification systems in our population

Macromolecular biosynthetic parameters and metabolic profile in different life stages of Leishmania braziliensis: Amastigotes as a functionally less active stage.

It was recently hypothesized that Leishmania amastigotes could constitute a semi-quiescent stage characterized by low replication and reduced metabolic activity. This concept developed with Leishmania (Leishmania) mexicana and Leishmania (Leishmania) major models might explain numerous clinical and sub-clinical features of Leishmania (Viannia) braziliensis infections, like reactivation of the disease, non-response to chemotherapy or asymptomatic infections. We compared here in vitro the proliferative capability of L. (V.) braziliensis amastigotes and promastigotes, assessed the expression of key molecular parameters and performed metabolomic analysis. We found that contrary to the highly proliferative promastigotes, amastigotes (axenic and intracellular) do not show evidence of extensive proliferation. In parallel, amastigotes showed a significant decrease of (i) the kDNA mini-circle abundance, (ii) the intracellular ATP level, (iii) the ribosomal components: rRNA subunits 18S and 28S α and ribosomal proteins RPS15 and RPL19, (iv) total RNA and protein levels. An untargeted metabolomic study identified clear differences between the different life stages: in comparison to logarithmic promastigotes, axenic amastigotes showed (a) a strong decrease of 14 essential and non-essential amino acids and eight metabolites involved in polyamine synthesis, (b) extensive changes in the phospholipids composition and (c) increased levels of several endogenous and exogenous sterols. Altogether, our results show that L. (V.) braziliensis amastigotes can show a phenotype with negligible rate of proliferation, a lower capacity of biosynthesis, a reduced bio-energetic level and a strongly altered metabolism. Our results pave the way for further exploration of quiescence among amastigotes of this species.

Alteraciones en la cavidad bucal en pacientes tratados con radioterapia de cabeza y cuello. Medellín, Colombia / Alterations found in the mouth of patients treated with head and neck radiotherapy. Medellin, Colombia

El objetivo del estudio fue determinar las alteraciones bucales en pacientes con cáncer de cabeza y cuello tratados con radioterapia y explorar la participación del Odontólogo en la atención de estos pacientes. Se realizó un estudio exploratorio en 52 pacientes que habían recibido más de 1,000 cGy de radiación. Se encuestó y realizó examen bucal a cada participante, tasa de flujo salivar estimulado y prueba de sabores. Los resultados muestran alta prevalencia de alteraciones bucales en pacientes con radiación acumulada entre 3,001 y 5,000 cGy. La boca seca (xerostomía) fue la alteración más sentida (78.8%). La tasa de secreción salivar total estimulada confirmó hiposalivación en el 82.7% de los pacientes. Se encontró una asociación estadísticamente significativa entre el lugar del cáncer (p < 0.01) y el tipo de tumor con la presencia de trismus (p < 0.05). La hiposialia se presentó más en los pacientes con tumores en estadio IV (50%) y en aquellos sometidos a tratamientos combinados (p < 0.05). Fue posible realizar la valoración periodontal al 50% de los pacientes, el 92% de ellos presentó periodontitis con pérdida de inserción principalmente severa y moderada. El 84.6% de los participantes manifestaron no haber sido remitidos a odontólogo antes o durante el tratamiento. Los hallazgos ratifican una alta frecuencia de alteraciones bucales en pacientes sometidos a tratamiento de radioterapia e inoportunidad de atención odontológica para prevenir o tratar estos efectos. Se alerta sobre la obligatoriedad de seguir protocolos de manejo del paciente oncológico, incluyendo valoración odontológica antes, durante y después del tratamiento respectivo.

The aim of the present study was to determine which oral alterations can be found in patients with head and neck cancer treated with radiotherapy as well as to explore dentist's involvement in treatment of these patients. An exploratory study was conducted in 52 patients who had previously received over 1,000 cGy radiation. A survey was undertaken as well as oral examination of each participant, in order to assess stimulated salivary flow and flavor tests. Results revealed high prevalence of oral alterations in patients with accumulated radiation of 3,001-5,000 cGy. Dry mouth (xerostomia) was the most frequently reported alteration (78.8%). Estimated total salivary secretion rate confirmed a state of hyposalivation in 82.7% of all patients. A statistically significant association was found between cancer location (p < 0.01) and type of tumor with presence of trismus (p < 0.05). Hyposialia was more frequently present in patients with stage IV tumors (50%) in those subjected to combined treatments (p < 0.05). Periodontal assessment was possible in 50% of all patients, Of this proportion, 92% exhibited periodontitis with mainly moderate to severe insertion loss; 84.6% of all participants reported not to have been remitted to dentists either before or after treatment. Findings support high frequency of oral alterations in patients subjected to radiotherapy treatment and dental care inappropriateness to prevent or treat these effects. An alert is raised with respect to the compulsiveness to follow treatment protocols for cancer patients, which should include dental evaluation before, during and after respective treatment.

Association of the Endobiont Double-Stranded RNA Virus LRV1 With Treatment Failure for Human Leishmaniasis Caused by Leishmania braziliensis in Peru and Bolivia.

Cutaneous and mucosal leishmaniasis, caused in South America by Leishmania braziliensis, is difficult to cure by chemotherapy (primarily pentavalent antimonials [Sb(V)]). Treatment failure does not correlate well with resistance in vitro, and the factors responsible for treatment failure in patients are not well understood. Many isolates of L. braziliensis (>25%) contain a double-stranded RNA virus named Leishmaniavirus 1 (LRV1), which has also been reported in Leishmania guyanensis, for which an association with increased pathology, metastasis, and parasite replication was found in murine models. Here we probed the relationship of LRV1 to drug treatment success and disease in 97 L. braziliensis-infected patients from Peru and Bolivia. In vitro cultures were established, parasites were typed as L. braziliensis, and the presence of LRV1 was determined by reverse transcription-polymerase chain reaction, followed by sequence analysis. LRV1 was associated significantly with an increased risk of treatment failure (odds ratio, 3.99; P = .04). There was no significant association with intrinsic Sb(V) resistance among parasites, suggesting that treatment failure arises from LRV1-mediated effects on host metabolism and/or parasite survival. The association of LRV1 with clinical drug treatment failure could serve to guide more-effective treatment of tegumentary disease caused by L. braziliensis.

Quantification of Leishmania (Viannia) Kinetoplast DNA in Ulcers of Cutaneous Leishmaniasis Reveals Inter-site and Inter-sampling Variability in Parasite Load.

BACKGROUND: Cutaneous leishmaniasis (CL) is a skin disease caused by the protozoan parasite Leishmania. Few studies have assessed the influence of the sample collection site within the ulcer and the sampling method on the sensitivity of parasitological and molecular diagnostic techniques for CL. Sensitivity of the technique can be dependent upon the load and distribution of Leishmania amastigotes in the lesion. METHODOLOGY/PRINCIPAL FINDINGS: We applied a quantitative real-time PCR (qPCR) assay for Leishmania (Viannia) minicircle kinetoplast DNA (kDNA) detection and parasite load quantification in biopsy and scraping samples obtained from 3 sites within each ulcer (border, base, and center) as well as in cytology brush specimens taken from the ulcer base and center. A total of 248 lesion samples from 31 patients with laboratory confirmed CL of recent onset (≤3 months) were evaluated. The kDNA-qPCR detected Leishmania DNA in 97.6% (242/248) of the examined samples. Median parasite loads were significantly higher in the ulcer base and center than in the border in biopsies (P<0.0001) and scrapings (P = 0.0002). There was no significant difference in parasite load between the ulcer base and center (P = 0.80, 0.43, and 0.07 for biopsy, scraping, and cytology brush specimens, respectively). The parasite load varied significantly by sampling method: in the ulcer base and center, the descending order for the parasite load levels in samples was: cytology brushes, scrapings, and biopsies (P<0.0001); in the ulcer border, scrapings had higher parasite load than biopsies (P<0.0001). There was no difference in parasite load according to L. braziliensis and L. peruviana infections (P = 0.4). CONCLUSION/SIGNIFICANCE: Our results suggest an uneven distribution of Leishmania amastigotes in acute CL ulcers, with higher parasite loads in the ulcer base and center, which has implications for bedside collection of diagnostic specimens. The use of scrapings and cytology brushes is recommended instead of the more invasive biopsy.

Relevance to Home Blood Pressure Monitoring Protocol of Blood Pressure Measurements Taken Before First- Morning Micturition and in the Afternoon / Relevância das Medidas da Pressão Arterial Pré-Micção Matinal e à Tarde em Protocolo de Monitorização Residencial da Pressão Arterial

Background: The importance of measuring blood pressure before morning micturition and in the afternoon, while working, is yet to be established in relation to the accuracy of home blood pressure monitoring (HBPM). Objective: To compare two HBPM protocols, considering 24-hour ambulatory blood pressure monitoring (wakefulness ABPM) as gold-standard and measurements taken before morning micturition (BM) and in the afternoon (AM), for the best diagnosis of systemic arterial hypertension (SAH), and their association with prognostic markers. Methods: After undergoing 24-hour wakefulness ABPM, 158 participants (84 women) were randomized for 3- or 5-day HBPM. Two variations of the 3-day protocol were considered: with measurements taken before morning micturition and in the afternoon (BM+AM); and with post-morning-micturition and evening measurements (PM+EM). All patients underwent echocardiography (for left ventricular hypertrophy - LVH) and urinary albumin measurement (for microalbuminuria - MAU). Result: Kappa statistic for the diagnosis of SAH between wakefulness-ABPM and standard 3-day HBPM, 3-day HBPM (BM+AM) and (PM+EM), and 5-day HBPM were 0.660, 0.638, 0.348 and 0.387, respectively. The values of sensitivity of (BM+AM) versus (PM+EM) were 82.6% × 71%, respectively, and of specificity, 84.8% × 74%, respectively. The positive and negative predictive values were 69.1% × 40% and 92.2% × 91.2%, respectively. The comparisons of intraclass correlations for the diagnosis of LVH and MAU between (BM+AM) and (PM+EM) were 0.782 × 0.474 and 0.511 × 0.276, respectively. Conclusions: The 3 day-HBPM protocol including measurements taken before morning micturition and during work in the afternoon showed the best agreement with SAH diagnosis and the best association with prognostic markers. .

Fundamentos: A importância das medidas da pressão arterial antes do ato miccional matinal e no período da tarde, durante atividades laborativas, na acurácia da monitoração residencial da pressão arterial (MRPA) não foi estabelecida. Objetivo: Comparar dois protocolos de MRPA, tendo como padrão-ouro a monitoração ambulatorial da pressão arterial de 24 horas (MAPA-vigília) e avaliando-se as medidas antes do ato miccional e à tarde, para o melhor diagnóstico de hipertensão arterial (HAS), e sua associação com marcadores prognósticos. Métodos: Após realizarem MAPA de 24 horas, os 158 participantes (84 mulheres) foram randomizados para realizar MRPA de três ou cinco dias com posterior crossover. Analisou-se o protocolo de três dias nas seguintes situações: aferições antes do ato miccional matinal e à tarde (AM+MT); e aferições após o ato miccional matinal e à noite (PM+MN). Todos os pacientes foram submetidos a ecocardiografia (hipertrofia ventricular esquerda - HVE) e a dosagem de albumina urinária (microalbuminúria - MAU). Resultados: A estatística kappa para diagnóstico de HAS entre MAPA-vigília e MRPA de três dias padrão, MRPA de três dias (AM + MT) e (PM + MN), e MRPA de cinco dias foi de 0,660, 0,638, 0,348 e 0,387, respectivamente. Os valores de sensibilidade de AM+MT versus PM+MN foram 82,6% × 71%, respectivamente, e os de especificidade foram 84,8% × 74%, respectivamente. Os valores preditivos positivos e negativos foram 69,1% × 40% e 92,2% × 91,2%, respectivamente. As comparações das correlações intraclasse para diagnósticos de HVE e MAU, entre AM+MT e PM+MN, foram 0,782 × 0,474 e 0,511 × 0,276, respectivamente. Conclusões: O protocolo de MRPA de três dias ...

Leishmanicidal compounds and potent PPARγ activators from Renealmia thyrsoidea (Ruiz & Pav.) Poepp. & Endl.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves and rhizomes of Renealmia thyrsoidea (Ruiz & Pav.) Poepp. & Endl. traditionally used in the Yanesha pharmacopoeia to treat skin infections such as leishmaniasis ulcers, or to reduce fever were chemically investigated to identify leishmanicidal compounds, as well as PPARγ activators. METHODS: Compounds were isolated through a bioassay-guided fractionation and their structures were determined via detailed spectral analysis. The viability of Leishmania amazonensis axenic amastigotes was assessed by the reduction of tetrazolium salt (MTT), the cytotoxicity on macrophage was evaluated using trypan blue dye exclusion method, while the percentage of infected macrophages was determined microscopically in the intracellular macrophage-infected assay. The CD36, mannose receptor (MR) and dectin-1 mRNA expression on human monocytes-derived macrophages was evaluated by quantitative real-time PCR. RESULTS: Six sesquiterpenes (1-6), one dihydrobenzofuranone (7) and four flavonoids (8-11) were isolated from the leaves. Alongside, two flavonoids (12-13) and five diarylheptanoids (14-18) were identified in the rhizomes. Leishmanicidal activity against Leishmania amazonensis axenic amastigotes was evaluated for all compounds. Compounds 6, 7, and 11, isolated from the leaves, showed to be the most active derivatives. Diarylheptanoids 14-18 were also screened for their ability to activate PPARγ nuclear receptor in macrophages. Compounds 17 and 18 bearing a Michael acceptor moiety strongly increased the expression of PPARγ target genes such as CD36, Dectin-1 and mannose receptor (MR), thus revealing interesting immunomodulatory properties. CONCLUSIONS: Phytochemical investigation of Renealmia thyrsoidea has led to the isolation of leishmanicidal compounds from the leaves and potent PPARγ activators from the rhizomes. These results are in agreement with the traditional uses of the different parts of Renealmia thyrsoidea.

In vitro evaluation of a soluble Leishmania promastigote surface antigen as a potential vaccine candidate against human leishmaniasis.

PSA (Promastigote Surface Antigen) belongs to a family of membrane-bound and secreted proteins present in several Leishmania (L.) species. PSA is recognized by human Th1 cells and provides a high degree of protection in vaccinated mice. We evaluated humoral and cellular immune responses induced by a L. amazonensis PSA protein (LaPSA-38S) produced in a L. tarentolae expression system. This was done in individuals cured of cutaneous leishmaniasis due to L. major (CCLm) or L. braziliensis (CCLb) or visceral leishmaniasis due to L. donovani (CVLd) and in healthy individuals. Healthy individuals were subdivided into immune (HHR-Lm and HHR-Li: Healthy High Responders living in an endemic area for L. major or L. infantum infection) or non immune/naive individuals (HLR: Healthy Low Responders), depending on whether they produce high or low levels of IFN-γ in response to Leishmania soluble antigen. Low levels of total IgG antibodies to LaPSA-38S were detected in sera from the studied groups. Interestingly, LaPSA-38S induced specific and significant levels of IFN-γ, granzyme B and IL-10 in CCLm, HHR-Lm and HHR-Li groups, with HHR-Li group producing TNF-α in more. No significant cytokine response was observed in individuals immune to L. braziliensis or L. donovani infection. Phenotypic analysis showed a significant increase in CD4+ T cells producing IFN-γ after LaPSA-38S stimulation, in CCLm. A high positive correlation was observed between the percentage of IFN-γ-producing CD4+ T cells and the released IFN-γ. We showed that the LaPSA-38S protein was able to induce a mixed Th1 and Th2/Treg cytokine response in individuals with immunity to L. major or L. infantum infection indicating that it may be exploited as a vaccine candidate. We also showed, to our knowledge for the first time, the capacity of Leishmania PSA protein to induce granzyme B production in humans with immunity to L. major and L. infantum infection.

[Recurrent diarrhea due to Cystoisopora belli in HIV/AIDS patients receiving HAART]. / Diarrea recurrente por Cystoisopora belli en pacientes con infección por VIH con TARGA.

The Cystoisospora belli, before denominated as Isospora belli, is the etiologic agent of cystoisosoporiasis, an opportunistic infection affecting immunocompromised patients, characterized by chronic diarrhea and weight loss. The incidence of chronic diarrhea for this agent, in HIV patients, has decreased considerably. This thanks to the advent of highly active antiretroviral therapy (HAART), which has improved the patient's immune response and decrease viral load. We present six cases of cystoisosoporiasis recurrent and refractory to treatment in HIV patients, who was being treated with with trimethoprim / sulfamethoxazole (TMP / SMX) orally as a prophylaxis. Five of these patients passed away due to the infection, despite of the fact that they had a good response to HAART (adequate increase in CD4 and viral load undetectable) and they had been treated with second line drugs.

Diarrea recurrente por Cystoisopora belli en pacientes con infección por VIH con TARGA / Recurrent diarrhea due to Cystoisopora belli in HIV/AIDS patients receiving HAART

La Cystoisospora belli, antes denominada Isospora belli, es el agente etiológico de la cystoisosporiasis, una infección oportunista que afecta a pacientes inmunodeprimidos, caracterizada por diarrea crónica y pérdida ponderal. La incidencia de diarrea crónica por este agente, en pacientes infectados por el VIH, ha disminuido considerablemente. Ello gracias al advenimiento de la terapia antirretroviral de gran actividad (TARGA), con la que se ha logrado mejorar la respuesta inmunológica del paciente y disminuir su carga viral. Se presentan seis casos de cystoisosporiasis recurrente y refractaria en pacientes con infección por VIH, en quienes se diagnosticó cystoisosporiasis pese a que previamente se encontraban recibiendo profilaxis con trimetropin/sulfametoxazol (TMP/SMX). Cinco de ellos evolucionaron de manera tórpida y fallecieron, a pesar de una buena respuesta al TARGA (adecuado incremento de CD4 y disminución de la carga viral hasta rangos indetectables), y de tratamiento con TMP/SMX por vía oral y otros medicamentos de segunda línea.

The Cystoisospora belli, before denominated as Isospora belli, is the etiologic agent of cystoisosoporiasis, an opportunistic infection affecting immunocompromised patients, characterized by chronic diarrhea and weight loss. The incidence of chronic diarrhea for this agent, in HIV patients, has decreased considerably. This thanks to the advent of highly active antiretroviral therapy (HAART), which has improved the patient’s immune response and decrease viral load. We present six cases of cystoisosoporiasis recurrent and refractory to treatment in HIV patients, who was being treated with with trimethoprim / sulfamethoxazole (TMP / SMX) orally as a prophylaxis. Five of these patients passed away due to the infection, despite of the fact that they had a good response to HAART (adequate increase in CD4 and viral load undetectable) and they had been treated with second line drugs.

Real-time PCR assay for detection and quantification of Leishmania (Viannia) organisms in skin and mucosal lesions: exploratory study of parasite load and clinical parameters.

Earlier histopathology studies suggest that parasite loads may differ between cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) lesions and between acute and chronic CL. Formal demonstration requires highly sensitive detection and accurate quantification of Leishmania in human lesional tissue. In this study, we developed a quantitative real-time PCR (qPCR) assay targeting minicircle kinetoplast DNA (kDNA) to detect and quantify Leishmania (Viannia) parasites. We evaluated a total of 156 lesion biopsy specimens from CL or ML suspected cases and compared the quantitative performance of our kDNA qPCR assay with that of a previously validated qPCR assay based on the glucose-6-phosphate dehydrogenase (G6PD) gene. We also examined the relationship between parasite load and clinical parameters. The kDNA qPCR sensitivity for Leishmania detection was 97.9%, and its specificity was 87.5%. The parasite loads quantified by kDNA qPCR and G6PD qPCR assays were highly correlated (r = 0.87; P < 0.0001), but the former showed higher sensitivity (P = 0.000). CL lesions had 10-fold-higher parasite loads than ML lesions (P = 0.009). Among CL patients, the parasite load was inversely correlated with disease duration (P = 0.004), but there was no difference in parasite load according to the parasite species, the patient's age, and number or area of lesions. Our findings confirm that CL and recent onset of disease (<3 months) are associated with a high parasite load. Our kDNA qPCR assay proved highly sensitive and accurate for the detection and quantification of Leishmania (Viannia) spp. in lesion biopsy specimens. It has potential application as a diagnostic and follow-up tool in American tegumentary leishmaniasis.

Comparative zymographic analysis of metallopeptidase of Leishmania (Viannia) peruviana and Leishmania (Viannia) braziliensis isolates from Peru.

American tegumentary leishmaniasis (ATL) in Peru is mainly associated with Leishmania (Viannia) peruviana and L. (V.) braziliensis. These parasites are genetically related, and their characterization as distinct species is controversial. Despite their genetic similarity, each species is associated with different clinical manifestations of ATL; L. (V.) peruviana causes only cutaneous leishmaniasis, whereas L. (V.) braziliensis can cause both cutaneous and mucocutaneous leishmaniasis. Because the primary cutaneous lesions caused by infection with these species are indistinguishable, it is necessary to develop a suitable method to differentiate them in order to prevent possible metastasis to oropharyngeal mucosa. In the present study, we investigated the proteolytic profile of L. (V.) peruviana and L. (V.) braziliensis isolates from Peru by zymographic analysis in SDS-PAGE copolymerized with gelatin. Enzymes were characterized according to their pH range of activity and sensitivity to distinct peptidase inhibitors. We observed that L. (V.) peruviana isolates displayed three proteolytic bands with molecular masses ranging from 55 to 80 kDa, whereas L. (V.) braziliensis isolates showed six proteolytic activities between 55 and 130 kDa. Using specific inhibitors, we determined that these proteolytic activities are due to metallopeptidases and present optimal activity between the pH range 5.5 and 10.0. Our results suggest that the expression of metallopeptidases in L. (V.) peruviana and L. (V.) braziliensis isolates from Peru is species-specific.

Non-invasive cytology brush PCR diagnostic testing in mucosal leishmaniasis: superior performance to conventional biopsy with histopathology.

BACKGROUND: Traditional methods of diagnosing mucosal leishmaniasis (ML), such as biopsy with histopathology, are insensitive and require collection of an invasive diagnostic specimen. METHODS: We compared standard invasive procedures including biopsy histopathology, biopsy PCR, and leishmanin skin test (LST) to a novel, non-invasive, cytology-brush based PCR for the diagnosis of ML in Lima, Peru. Consensus reference standard was 2/4 tests positive, and outcome measures were sensitivity and specificity. Leishmania species identification was performed by PCR-based assays of positive specimens. RESULTS: Twenty-eight patients were enrolled, 23 of whom fulfilled criteria for a diagnosis of ML. Sensitivity and specificity of biopsy with histopathology were 21.7% [95% CI 4.9-38.5%] and 100%; 69.6% [95% CI 50.8-88.4%] and 100% for LST; 95.7% [95% CI 87.4-100%] and 100% for biopsy PCR; and 95.7% [95% CI 87.4-100%] and 90% [95% CI 71.4-100%] for cytology brush PCR using both Cervisoft® and Histobrush® cervical cytology brushes. Represented species identified by PCR-RFLP included: L. (V). braziliensis (n = 4), and L. (V). peruviana (n = 3). CONCLUSIONS: Use of commercial grade cytology brush PCR for diagnosis of ML is sensitive, rapid, well tolerated, and carries none of the risks of invasive diagnostic procedures such as biopsy. Further optimization is required for adequate species identification. Further evaluation of this method in field and other settings is warranted.

Multilocus genotyping reveals a polyphyletic pattern among naturally antimony-resistant Leishmania braziliensis isolates from Peru.

In order to understand the epidemiological dynamics of antimonial (Sb(V)) resistance in zoonotic tegumentary leishmaniasis and its link with treatment outcome, we analyzed the population structure of 24 Peruvian Leishmania braziliensis clinical isolates with known in vitro antimony susceptibility and clinical phenotype by multilocus microsatellite typing (14 microsatellite loci). The genetic variability in the Peruvian isolates was high and the multilocus genotypes were strongly differentiated from each other. No correlation was found between the genotypes and in vitro drug susceptibility or clinical treatment outcome. The finding of a polyphyletic pattern among the Sb(V)-resistant L. braziliensis might be explained by (i) independent events of drug resistance emergence, (ii) sexual recombination and/or (iii) other phenomena mimicking recombination signals. Interestingly, the polyphyletic pattern observed here is very similar to the one we observed in the anthroponotic Leishmania donovani (Laurent et al., 2007), hereby questioning the role of transmission and/or chemotherapeutic drug pressure in the observed population structure.