Multiantigen print immunoassay (MAPIA) for the diagnosis of neurocysticercosis: a single-center diagnostic optimization and accuracy study in Lima, Peru.

Neurocysticercosis (NCC) is the most common helminthic infection of the human central nervous system. The antibody detection assay of choice is the enzyme-linked immunoelectrotransfer blot assay using lentil-lectin purified parasite antigens (LLGP-EITB, Western blot), an immunoassay with exceptional performance in clinical samples. However, its use is mainly restricted to a few research laboratories because the assay is labor-intensive and requires sophisticated equipment, expertise, and large amounts of parasite material for preparation of reagents. We report a new immunoprint assay (MAPIA) that overcomes most of these barriers. We initially compared the performance of five different antigen combinations in a subset of defined samples in the MAPIA format. After selecting the best-performing assay format (a combination of rGP50 + rT24H + sTs14 antigens), 148 archived serum samples were tested, including 40 from individuals with parenchymal NCC, 40 with subarachnoid NCC, and 68 healthy controls with no evidence of neurologic disease. MAPIA using three antigens (rGP50 + rT24H + sTs14) was highly sensitive and specific for detecting antibodies in NCC. It detected 39 out of 40 (97.5%) parenchymal NCC cases and 40/40 (100%) subarachnoid cases and was negative in 67 out of 68 (98.53%) negative samples. MAPIA using three recombinant and synthetic antigens is a simple and economical tool with a performance equivalent to the LLGP-EITB assay for the detection of specific antibodies to NCC. The MAPIA overcomes existing barriers to adoption of the EITG LLGP and is a candidate for worldwide use.

Neurocysticercosis. A frequent cause of seizures, epilepsy, and other neurological morbidity in most of the world.

Neurocysticercosis is endemic in most of the world and in endemic areas it accounts for approximately 30% of cases of epilepsy. Appropriate diagnosis and management of neurocysticercosis requires understanding the diverse presentations of the disease since these will vary in regards to clinical manifestation, sensitivity of diagnostic tests, and most importantly, therapeutic approach. This review attempts to familiarize tropical neurology practitioners with the diverse types of neurocysticercosis and the more appropriate management approaches for each.

Decrypting the sulfur cycle in oceanic oxygen minimum zones.

Here we present ecophysiological studies of the anaerobic sulfide oxidizers considered critical to cryptic sulfur cycling in oceanic oxygen minimum zones (OMZs). We find that HS- oxidation rates by microorganisms in the Chilean OMZ offshore from Dichato are sufficiently rapid (18 nM h-1), even at HS- concentrations well below 100 nM, to oxidize all sulfide produced during sulfate reduction in OMZs. Even at 100 nM, HS- is well below published half-saturation concentrations and we conclude that the sulfide-oxidizing bacteria in OMZs (likely the SUP05/ARTIC96BD lineage of the gammaproteobacteria) have high-affinity (>105 g-1 wet cells h-1) sulfur uptake systems. These specific affinities for sulfide are higher than those recorded for any other organism on any other substrate. Such high affinities likely allow anaerobic sulfide oxidizers to maintain vanishingly low sulfide concentrations in OMZs driving marine cryptic sulfur cycling. If more broadly distributed, such high-affinity sulfur biochemistry could facilitate sulfide-based metabolisms and prominent S-cycles in many other ostensibly sulfide-free environments.

Initial Experience in the Use of Novel Auto-expandable Metal Ureteral Stent in the Treatment of Ureter Stenosis in Kidney Transplanted Patients.

INTRODUCTION: Ureter stenosis in renal transplantation patients is a relatively frequent complication that negatively conditions graft evolution. The use of ureteral stents is a valid treatment alternative to the use of double-J catheters in patients for whom surgery is not contemplated or after surgical recurrence. We present our initial experience with five patients treated using this technique. MATERIALS AND METHODS: We describe a total of five patients with ureteral stenosis after renal transplantation who were treated using ureteral stent model UVENTA (Taewoong Medical, Seoul, Korea) in our center. The median follow-up was 18 months (range, 4 to 38 months). We describe the clinical history of patients and previous treatments on ureteral stenosis. The technical procedure of placement is described. The clinical course is analyzed by measurement of renal function and imaging tests, as well as post-stent complications. Survival of the renal graft is evaluated. RESULTS: The procedure could be completed in all patients without complications. The technique was effective in all patients, with correction of creatinine value and hydronephrosis during the renal ultrasound test. One patient suffered a urinary tract infection episode associated with the use of the ureteral stent. One patient suffered the loss of the renal graft secondary to the development of cryoglobulins. One hundred percent of the ureteral stents are functioning as of the writing of this article. CONCLUSIONS: In renal transplantation patients with ureter stenosis, metallic stents are a useful technique with low morbidity and associated complications.

What Is the Influence of Both Risk Donor and Risk Receiver on Simultaneous Pancreas-Kidney Transplantation?

INTRODUCTION: Some factors affect the pancreas of a marginal donor, and although their influence on graft survival has been determined, there is an increasing consensus to accept marginal organs in a controlled manner to increase the pool of organs. Certain factors related to the recipient have also been proposed as having negative influence on graft prognosis. The objective of this study was to analyze the influence of these factors on the results of our simultaneous pancreas-kidney (SPK) transplantation series. MATERIALS AND METHODS: Retrospective analysis of 126 SPK transplants. Donors and recipients were stratified in an optimal group (<2 expanded donor criteria) and a risk group (≥2 criteria). A pancreatic graft survival analysis was performed using a Kaplan-Meier test and log-rank test. Prognostic variables on graft survival were studied by Cox regression. Postoperative complications (graded by Clavien classification) were compared by χ2 test or Fisher test. RESULTS: Median survival of pancreas was 66 months, with no significant difference between groups (P > .05). Multivariate analysis showed risk factors to be donor age, cold ischemia time, donor body mass index, receipt body mass index, and receipt panel-reactive antibody. CONCLUSIONS: In our series, the use of pancreatic grafts from donors with expanded criteria is safe and has increased the pool of grafts. Different variables, both donor and recipient, influence the survival of the pancreatic graft and should be taken into account in organ distribution systems.

Intraoperative Heparinization During Simultaneous Pancreas-Kidney Transplantation: Is It Really Necessary?

INTRODUCTION: The use of intraoperative sodium heparin during simultaneous pancreas-kidney transplantation (SPKT) remains as a routine practice in some referral centers to minimize pancreatic graft thrombosis rate. One of its disadvantages is the theoretical increased risk of postoperative bleeding. In our center, we have abandoned its use since 2011. MATERIALS AND METHODS: We performed a retrospective analysis among 198 SPKTs performed in our center between the years 1989 and 2017. The variables of our study were vascular thrombosis of the pancreatic graft and hemoperitoneum and upper gastrointestinal bleeding in the mediate postoperative period (up to 2 months after the transplant). We compared these results between SPKT recipients who had undergone intraoperative heparinization (n = 157) and those who had not (n = 51). To avoid bias, a second comparison was performed using propensity score matching on the following characteristics: sex, recipient age, recipient body mass index, cold ischemia time, preoperative hemodialysis or peritoneal dialysis, time of diabetes, and Pancreas Donor Risk Index. Student t test or Mann-Whitney U test was used for intergroup comparisons of quantitative variables where appropriate, whereas χ2 or Fisher exact test was used to compare categorical data. RESULTS: No statistically significant differences were found when comparing the use of intraoperative heparin, even after the homogenization of both groups. CONCLUSIONS: In our experience, intraoperative heparinization during SPKT surgery was not useful because it did not significantly decrease the graft thrombosis rate, and its withdrawal did not enhance hemoperitoneum or upper gastrointestinal bleeding postoperative rates.

Pancreas Donor Hypernatremia: Is it Really a Risk Factor for Simultaneous Pancreas-kidney Transplantation?

INTRODUCTION: Solid organ donor hypernatremia has been classically reported to be a risk factor for cell lysis and graft damage. National criteria for pancreatic donation consider severe hypernatremia (sodium level more than 160 mEq/L) to be relative exclusion criteria. The aim of our study is to review the postoperative outcomes of our simultaneous pancreas-kidney transplantation (SPKT) sample in terms of pancreatic fistula, intra-abdominal abscesses, pancreatitis, pancreas graft thrombosis, early pancreatectomy, and reoperation rates regarding different ranges of donor sodium levels. MATERIAL AND METHODS: We performed a retrospective analysis among 161 SPKTs performed in our center between the years 2001 and 2017. We compared the aforementioned postoperative variables in two situations: 1) Whether the donor pancreas sodium levels were inferior to 149 mEq/L, or equal to or greater than 150 mEq/L; and 2) If they had severe hypernatremia (considering sodium levels greater than or equal to 160 mEq/L as threshold) or not. To ensure the comparability of the groups, a second comparison was performed on new samples after using propensity score matching. A Student t test or Mann-Whitney U test was used for intergroup comparisons of quantitative variables where appropriate, whereas a χ2 test or Fisher's exact test was used to compare categorical data. RESULTS: No statistically significant differences were found between the groups that relate high donor serum sodium levels with the morbidity variables included in our study or with early pancreatic graft loss. CONCLUSIONS: In our cohort, early postoperative main morbidity and pancreas graft loss of SPKT recipients do not differ significantly regarding donor serum sodium levels.

Graft Survival in Patients Who Received Second Allograft, Comparing Those With or Without Previous Failed Allograft Nephrectomy.

INTRODUCTION: Nowadays, the number of patients receiving a second graft is growing, and the management of failed grafts is still controversial. OBJECTIVE: Our objective was to analyze the influence of graft nephrectomy on graft and patient survival. MATERIALS AND METHODS: We retrospectively evaluated the demographic features and graft outcomes of 63 recipients who received second allografts between August 1985 and April 2013. They were divided into two groups: group A, those who underwent nephrectomy of failed graft (n = 21, 33.3%), and group B, those whose failed graft was retained (n = 42, 66.6%). χ2 and Mann-Whitney U tests were used to compare demographic characteristics and graft features in both groups. Kaplan-Meier test was used to analyze graft and patient survival. Finally, univariate and multivariate analysis was done using Cox regression. RESULTS: Demographic characteristics of donor and receptors were similar in both groups. Overall panel-reactive antibody (P = .040) showed statistically significant differences between groups (72.0 ± 25.3 in group A and 54.8 ± 30.0 in group B). Hemodialysis duration was longer in group A (P = .023, 112.2 ± 72.8 vs 70.9 ± 66.9 months). The percentage of patients who had delayed graft function was higher in group A (58.8% vs 27.3%, P = .029). Kaplan-Meier test found no differences between groups (P = .344); group A, 107.4 months (95% confidence interval [CI] 74.0 to 140.8) and group B, 82.7 months (95% CI 62.5 to 102.8). We found no differences in terms of patient survival (P = .798) with the Kaplan-Meier test. In group A, patient survival was 164.5 months (CI 137.7 to 191.31) and in group B, 152.0 months (95% CI 125.5 to 178.5). CONCLUSIONS: Failed graft nephrectomy did not show a negative impact on graft and patient survival.

Influence of Donor and Recipient Ages in Survival of Simultaneous Pancreas-Kidney Transplantation.

INTRODUCTION: Simultaneous pancreas-kidney transplantation (SPKT) is a well treatment for patients with insulin-dependent diabetes and end-stage renal disease. Donor age is a barrier to the acceptance of organs. Age matching has been extensively studied in kidney transplantation; however, there are no studies in graft survival after SPKT. We aimed to study the combined influence of the ages of the donors and recipients in graft survival after SPKT. MATERIAL AND METHODS: Donors and recipients are classified as younger (age <40 years) or older (age ≥40 years). There were four study groups (young-young, young-old, old-young, and old-old). They were evaluated retrospectively for demographic and clinical characteristics of donors and recipients and the long-term survival between 2001 and 2012 of kidney pancreas transplantation patients at our center. RESULTS: A total of 115 transplantations were performed. The four groups had 55 young-young, 40 young-old, 10 old-young, and 10 old-old patients. Serious complications occurred in 32%, 42%, 30%, and 40%, respectively, and deaths were 2%, 5%, 0%, and 20%, respectively, in the groups. Pancreas graft survival at 3 years for each group was 80%, 87, 5%, 90%, and 60%, respectively, and kidney graft survival was 92.7%, 90%, 90%, and 70%, respectively. Panel-reactive antibodies (PRAs) >30% were associated with poor graft survival, and serious postoperative complications associated with poor pancreas-kidney graft survival. CONCLUSIONS: In conclusion, both younger and older recipients show excellent long-term graft and patient survival after SPKTs from younger donors. We recommended that older-recipient SPKT be transplanted from younger donors because older recipients who have been transplanted from older donors had decreased survival.

Validation of the Pancreatic Donor Risk Index in Simultaneous Pancreas-Kidney Transplantation Performed in Córdoba Hospital From 2000 to 2015.

BACKGROUND: The Pancreatic Donor Risk Index (PDRI) was developed in 2010 in the United States to predict graft survival after pancreas transplantation, based on donor characteristics and logistical and technical conditions. The aim of the study was to validate the utility of PDRI as a pancreas allograft survival predictor in simultaneous pancreas-kidney transplantation (SPK) transplants performed in our hospital between 2000 and 2015. METHODS: This retrospective analysis of 126 SPK transplants was performed by the same surgical team from the years 2000 to 2015. Donor variables that are integrated in the PDRI were calculated (age, sex, race, creatinine serum levels, body mass index, height, cold ischemia time, cause of death, type of pancreas transplant). Pancreatic graft survival at 1 and 5 years was calculated by use of the Kaplan-Meier test. Comparison of survival curves between PDRI risk quartiles was calculated by use of the log-rank test. Association between graft survival and variables integrating the PDRI was calculated by use of univariate Cox regression analysis. RESULTS: Log-rank analysis found no statistically significant association between global graft survival and PDRI quartiles. Univariate Cox regression analysis showed a statistically significant association between graft survival and cold ischemia time (P < .05). CONCLUSIONS: PDRI was not a useful tool to predict pancreatic graft outcomes in a Spanish reference population.

Recombinant protein production data after expression in the bacterium Escherichia coli.

Fusion proteins have become essential for the expression and purification of recombinant proteins in Escherichia coli. The metal-binding protein CusF has shown several features that make it an attractive fusion protein and affinity tag: "Expression and purification of recombinant proteins in Escherichia coli tagged with the metal-binding protein CusF" (Cantu-Bustos et al., 2016 [1]). Here we present accompanying data from protein expression experiments; we tested different protein tags, temperatures, expression times, cellular compartments, and concentrations of inducer in order to obtain soluble protein and low formation of inclusion bodies. Additionally, we present data from the purification of the green fluorescent protein (GFP) tagged with CusF, using Ag(I) metal affinity chromatography.

Expression and purification of recombinant proteins in Escherichia coli tagged with the metal-binding protein CusF.

Production of recombinant proteins in Escherichia coli has been improved considerably through the use of fusion proteins, because they increase protein solubility and facilitate purification via affinity chromatography. In this article, we propose the use of CusF as a new fusion partner for expression and purification of recombinant proteins in E. coli. Using a cell-free protein expression system, based on the E. coli S30 extract, Green Fluorescent Protein (GFP) was expressed with a series of different N-terminal tags, immobilized on self-assembled protein microarrays, and its fluorescence quantified. GFP tagged with CusF showed the highest fluorescence intensity, and this was greater than the intensities from corresponding GFP constructs that contained MBP or GST tags. Analysis of protein production in vivo showed that CusF produces large amounts of soluble protein with low levels of inclusion bodies. Furthermore, fusion proteins can be exported to the cellular periplasm, if CusF contains the signal sequence. Taking advantage of its ability to bind copper ions, recombinant proteins can be purified with readily available IMAC resins charged with this metal ion, producing pure proteins after purification and tag removal. We therefore recommend the use of CusF as a viable alternative to MBP or GST as a fusion protein/affinity tag for the production of soluble recombinant proteins in E. coli.

Instability of an aromatic amine in fatty food and fatty food simulant: characterisation of reaction products and prediction of their toxicity.

It is a well-known fact that amines are not stable in food of a fatty nature. In this study the synthesis and characterisation of the products obtained as a result of the reaction of amines in a fatty medium are reported. Based on the well-known reactions among amines and acid and esters groups, two novel compounds were synthesised using m-xylylenediamine (mXDA), a primary diamine widely used as monomer in the manufacture of food contact materials and two fatty acids, oleic acid and palmitic acid, which occur in most fats. The resulting compounds were two molecules belonging to the family of fatty acid amides, dioleamide and dipalmitamide. A complete characterisation of both products was carried out employing several techniques such as infrared spectroscopy, (1)H- and (13)C-NMR spectroscopy, electron ionisation mass spectrometry, LC-MS/MS and UV spectrometry. The results obtained by the different techniques were well correlated. In the second part of the work, the formation of these compounds in real samples was evaluated. For this purpose a certain volume of olive oil was spiked with a known amount of mXDA. Olive oil was selected as a fatty medium since it is a widely consumed food and additionally is used as a fatty food simulant in migration studies of food contact materials. A method was developed to extract the fatty acid amides from the fatty matrix, which were then identified by LC-MS/MS. The toxicity of the synthesised compounds was predicted using a toxicity estimation software tool.

Recurrencia luego del tratamiento conservador en cáncer de mama: su relación con la edad y factores inmunohistoquímicos / Recurrence after breast cancer conservative treatment: relationship with age and immunohistochemical factors

La estrategia terapéutica para los tumores de mama en estadios tempranos ha sido establecida desde fines del siglo XX; es entonces que al tratamiento quirúrgico conservador y la radioterapia se sumó el ganglio centinela y el tratamiento adyuvante, de acuerdo a parámetros pre- y posoperatorios. Sin embargo, existe en todas las series una tasa de recurrencia local, y se ha podido determinar en parte cuáles son los factores de la paciente, anatómicos e histológicos, que predisponen a tal evento, a partir de hallazgos posoperatorios.También existe una tasa de enfermedad regional y a distancia, derivando en la necesidad de identificar factores de predicción que puedan sugerir la necesidad de una terapéutica más agresiva en pacientes susceptibles. Por otra parte, la clasificación molecular de los tumores de mama descripta por Perou y cols. pone en evidencia características inherentes a cada tumor, permitiendo establecer subtipos con implicancia clínica. A partir del análisis de nuestra estadística, en la que presentamos pacientes con cáncer de mama inicial que han recibido un tratamiento conservador, los objetivos planteados son: Presentar las tasas de eventos locales, regionales, a distancia y sobrevida luego de 15 años de experiencia en tratamiento conservador, y determinar si existe en nuestra serie una relación entre estos eventos, la edad de las pacientes y la clasificación molecular de los tumores. Establecer a partir de los resultados su implicancia a nivel asistencial.

Simultaneous chromatographic analysis of photoinitiators and amine synergists in food contact materials.

Photoinitiators (PIs) are components of UV-cured inks widely used in printing of food packaging. These substances can migrate into food and may be a hazard to human health. High-performance liquid chromatography with diode-array detection (HPLC-DAD) has been used for analysis of PIs and amine synergists in food packaging. Analysis was performed with a Kromasil C18 column (250 mm × 3.2 mm i.d., 5 µm particle size) with a binary mobile phase gradient prepared from acetonitrile and Milli-Q water. The flow rate was 0.5 mL min(-1). The method enables separation of fourteen PIs and amine synergists in a single run. The method was validated for linearity, repeatability, and limits of detection and quantification. Excellent sensitivity (LODs ≤ 1.56 µg dm(2)) and appropriate repeatability (RSD (n = 10) <0.9%) were achieved. Different types of food packaging material including plastic films, cardboard, and cans were analyzed and PIs were detected in 47% of the samples tested (n = 17). Positive samples were confirmed by use of LC-MS-MS in positive electrospray ionization (ESI) mode.

Revisión de las esofagitis eosinofílicas en un hospital secundario en los últimos 4 años / Review of eosinophilic esophagitis in a secondary hospital in the last 4 years

Introducción. La esofagitis eosinofilica, es una inflamación crónica del esófago con una importante infiltración mucosa eosinofílica. Nuestro objetivo fue revisar las endoscopias con diagnóstico anatomopatológico de esofagitis eosinofílica y estudiar las características y evolución de los pacientes. Material y métodos. Estudio descriptivo retrospectivo, mediante revisión de informes clínicos desde enero del 2008 a diciembre de 2012 de pacientes pediátricos a los que se realizó estudio endoscópico con diagnóstico de esofagitis cosinofílica. El análisis se realizó mediante programa SPSS19. Resultados. Se registraron 13 pacientes. El 85% son varones. El síntoma principal fue la disfagia. La mayoría presentaban antecedentes personales y/o familiares de alergia. En el 100% se observaron > de 20 eosinófilos por campo de gran aumento. Los fármacos más usado fueron fluticasona y budesonida. En más de la mitad de los pacientes se realizó control endoscópico. Discusión y conclusiones. La esofagitis eosinofílica supone el 14,07% de las endoscopias realizadas en nuestro hospital en el tiempo registrado. Aunque es una patología poco frecuente, la falta de estudios sobre el tratamiento y pronóstico de la enfermedad hace necesaria la puesta en marcha de un protocolo de actuación que unifique el manejo de estos pacientes (AU)

Introduction. Eosinophilic esophagitis is a chronic inflammation of the esophageal mucosa with significant eosinophilic infiltration. Our objective was to review the endoscopy with pathological diagnosis of eosinophilic esophagitis and study the characteristics and patient outcomes. Material and methods. A retrospective review of clinical reports through form January 2008 to December 2012 pediatric patients undergoing endoscopy was performed with a diagnosis of eosinophilic esophagitis. The analysis was performed using SPSS 19 program. Results. 13 patients were recorded. 85% are male. The main symptom was dysphagia. Most families had a history of allergy. In 100% were observed > 20 eosinophils per high power field. The most commonly used drugs were fluticasome and budesonide. In more than half of patients endoscopic control was performed. Discussion and conclusions. Eosinophilic esophagitis represent 14,07% of endoscopies perfomed in our hospital at the time recorded. Although it is an uncommon condition, the lack of studies on the treatment and prognosis of the disease requires the implementation of a protocol that unifies the management of these patients (AU)

Características epidemiológicas del cáncer de mama en el área metropolitana de Buenos Aires y La Plata. Estudio de una serie de 4.041 casos del Registro de Cáncer de Mama (RCM) / Epidemiological aspects of breast cancer in metropolitana area of Buenos Aires and La Plata. Study of 4.041 patients of breast cancer registry (RCM)

Introducción: En la Argentina se estima que se detectan más de 18.000 casos nuevos de cáncer de mama por año; sin embargo, en nuestro país no contamos con registros de dicha patología que posean poder estadístico adecuado. Por este motivo, desde la Sociedad Argentina de Mastología surgió el interés de crear una base de datos. Aquí presentamos el análisis de los datos recabados de la misma luego de 2 años de su implementación en distintos centros de atención de Mastología del Area Metropolitana de la Ciudad de Buenos Aires (AMBA). Objetivo: Describir las características epidemiológicas de las pacientes atendidas en los centros participantes, así como la presentación clínica, estadio y tratamiento instaurado. Material y método: Estudio descriptivo, de corte transversal, donde se analizaron los datos ingresados hasta el día 30/09/2012, en la base de web "registro de cáncer de mama" (RCM), de 4.041 pacientes. Dichos datos fueron ingresados por los dieciséis centros participantes hasta ese momento, todos ellos ubicaodos en el AMBA y la ciudad de la Plata. Resultados: El 99,6% de los pacientes fue de sexo femenino, con una media de edad 57,7±13,2 años. Se registró una media de edad de aparición para la menarca de 12,5±1,4 años y de 48,6±4,1 años para la menopausia. El 68,8% de las pacientes eran posmenopáusicas. El 20,3% de las pacientes era nulípara. El 19,3% presentó el antecedente heredo-familiar de cáncer de mama. El principal motivo de consulta registrado fue el de autodetección sospechosa 54,3%. La distribución del estadio clínico fue la siguiente: estadio 0 11,6%; estadio I 40,6%; estadio II 36,5%; estadio III 10,3% y estadio IV 1,0%. En el 69,4% se realizó resección sectorial como cirugía mamaria. Antes del año 2000 predominó la linfadenectomía axilar (67,9%), luego de esta fecha la frecuencia de indicación del ganglio centinela pasó a aplicarse al 63,6%. (AU)