Population pharmacokinetics of benznidazole in neonates, infants and children using a new pediatric formulation.

BACKGROUND: There is a major need for information on pharmacokinetics (PK) of benznidazole (BNZ) in children with Chagas disease (CD). We conducted a multicentre population PK, safety and efficacy study in children, infants and neonates with CD treated with BNZ (formulated in 100 mg tablets or 12.5 mg dispersible tablets, developed by the pharmaceutical company LAFEPE, in a collaboration with DNDi). METHODS: 81 children 0-12 years old were enrolled at 5 pediatric centers in Argentina. Diagnosis of T. cruzi infection was confirmed by direct microscopic examination, or at least two positive conventional serological tests. Subject enrolment was stratified by age: newborns to 2 years (minimum of 10 newborns) and >2-12 years. BNZ 7.5 mg/kg/d was administered in two daily doses for 60 days. Five blood samples per child were obtained at random times within pre-defined time windows at Day 0 at 2-5 h post-dose; during steady state, one sample at Day 7 and at Day 30; and two samples at 12-24 h after final BNZ dose at Day 60. The primary efficacy endpoint was parasitological clearance by qualitative PCR at the end of treatment. RESULTS: Forty-one (51%) patients were under 2 years of age (including 14 newborns <1 month of age). Median age at enrolment was 22 months (mean: 43.2; interquartile range (IQR) 7-72 months). The median measured BNZ Cmax was 8.32 mg/L (IQR 5.95-11.8; range 1.79-19.38). Median observed BNZ Cmin (trough) concentration was 2 mg/L (IQR 1.25-3.77; range 0.14-7.08). Overall median simulated Css was 6.3 mg/L (IQR 4.7-8.5 mg/L). CL/F increased quickly during the first month of postnatal life and reached adult levels after approximately 10 years of age. Negative qPCR was observed at the end of treatment in all 76 patients who completed the treatment. Five patients discontinued treatment (3 due to AEs and 2 due to lack of compliance). CONCLUSION: We observed lower BNZ plasma concentrations in infants and children than those previously reported in adults treated with comparable mg/kg doses. Despite these lower concentrations, pediatric treatment was well tolerated and universally effective, with a high response rate and infrequent, mild AEs. TRIAL REGISTRATION: Registered in clinicaltrials.gov #NCT01549236.

Efficacy and Safety of Nifurtimox in Pediatric Patients with Chagas Disease: Results at 4-Year Follow-Up in a Prospective, Historically Controlled Study (CHICO SECURE).

Nifurtimox is recommended for the treatment of Chagas disease; however, long-term follow-up data are scarce. This prolonged follow-up phase of the prospective, historically controlled, CHICO clinical trial evaluated seronegative conversion in pediatric patients aged <18 years with Chagas disease who were followed for 4 years after nifurtimox treatment. Patients were randomly assigned 2:1 to nifurtimox 60-day or 30-day regimens comprising 10 to 20 mg/kg/day for patients aged <12 years and body weight <40 kg, and 8 to 10 mg/kg/day for those aged ≥12 years and body weight ≥40 kg. Anti-Trypanosoma cruzi antibodies decreased during the study period, achieving seronegative conversion in 16 (8.12%) and 8 (8.16%) patients in the 60-day and 30-day nifurtimox regimens, respectively, with corresponding incidence rates per 100 patients/year of seronegative conversion of 2.12 (95% confidence interval [CI]: 1.21 to 3.45) and 2.11 (95% CI: 0.91 to 4.16). Superiority of the 60-day nifurtimox regimen was confirmed by the lower limit of the 95% CI being higher than that (0%) in a historical placebo control group. Children aged <2 years at baseline were more likely to reach seronegative conversion during the 4-year follow-up than older children. At any annual follow-up visit, >90% of evaluable patients had persistently negative quantitative PCR results for T. cruzi DNA. No adverse events potentially related to treatment or caused by protocol-required procedures were documented for either treatment regimen. This study confirms the effectiveness and safety of a pediatric formulation of nifurtimox administered in an age- and weight-adjusted regimen for 60 days to treat children with Chagas disease.

Large-Size Subunit Catalases Are Chimeric Proteins: A H2O2 Selecting Domain with Catalase Activity Fused to a Hsp31-Derived Domain Conferring Protein Stability and Chaperone Activity.

Bacterial and fungal large-size subunit catalases (LSCs) are like small-size subunit catalases (SSCs) but have an additional C-terminal domain (CT). The catalytic domain is conserved at both primary sequence and structural levels and its amino acid composition is optimized to select H2O2 over water. The CT is structurally conserved, has an amino acid composition similar to very stable proteins, confers high stability to LSCs, and has independent molecular chaperone activity. While heat and denaturing agents increased Neurospora crassa catalase-1 (CAT-1) activity, a CAT-1 version lacking the CT (C63) was no longer activated by these agents. The addition of catalase-3 (CAT-3) CT to the CAT-1 or CAT-3 catalase domains prevented their heat denaturation in vitro. Protein structural alignments indicated CT similarity with members of the DJ-1/PfpI superfamily and the CT dimers present in LSCs constitute a new type of symmetric dimer within this superfamily. However, only the bacterial Hsp31 proteins show sequence similarity to the bacterial and fungal catalase mobile coil (MC) and are phylogenetically related to MC_CT sequences. LSCs might have originated by fusion of SSC and Hsp31 encoding genes during early bacterial diversification, conferring at the same time great stability and molecular chaperone activity to the novel catalases.

The Small Metal-Binding Protein SmbP Simplifies the Recombinant Expression and Purification of the Antimicrobial Peptide LL-37.

(1) Background: The cathelicidin peptide LL-37 is a prominent molecule with many biological activities, including antimicrobial. Due to its importance, here, we describe the production of LL-37 tagged with SmbP, a relatively new carrier protein that improves the production of recombinant proteins and peptides in Escherichia coli. We present an alternative method for the rapid expression, purification, and antimicrobial evaluation of LL-37, that involves only one purification step. (2) Methods: A DNA construct of SmbP_LL-37 was transformed into E. coli BL21(DE3); after overnight expression, the protein was purified directly from the cell lysate using immobilized metal-affinity chromatography. SmbP_LL-37 was treated with Enterokinase to obtain the free LL-37 peptide. The antimicrobial activity of both SmbP_LL-37 and free LL-37 was determined using the colony forming unit assay method. (3) Results: SmbP_LL-37 was observed in the soluble fraction of the cell lysate; after purification with IMAC, protein gel electrophoresis, and analysis by ImageJ, it showed 90% purity. A total of 3.6 mg of SmbP_LL-37 was produced from one liter of cell culture. SmbP_LL-37 and free LL-37 both showed inhibition activity against Staphylococcus aureus and Escherichia coli. (4) Conclusions: The SmbP fusion protein is a valuable tool for producing biologically-active LL-37 peptide. The production method described here should be of interest for the expression and purification of additional cationic peptides, since it cuts the purification time considerably prior to determination of antimicrobial activity.

Serological response to nifurtimox in adult patients with chronic Chagas disease: An observational comparative study in Argentina.

Nifurtimox is indicated in Chagas disease but determining its effectiveness in chronic disease is hindered by the length of time needed to demonstrate negative serological conversion. We manually reviewed long-term follow-up data from hospital records of patients with chronic Chagas disease (N = 1,497) in Argentina diagnosed during 1967-1980. All patients were aged ≥18 years at diagnosis and were either treated with nifurtimox (n = 968) or received no antitrypanosomal treatment (n = 529). The primary endpoint was negative seroconversion (the "event"), defined as a change from positive to negative in the serological or parasitological laboratory test used at diagnosis. Time to event was from baseline visit to date of endpoint event or censoring. The effectiveness of nifurtimox versus no treatment was estimated with Cox proportional hazard regression using propensity scores with overlap weights to calculate the hazard ratio and 95% confidence interval. The nifurtimox group was younger than the untreated group (mean, 32.4 vs. 40.3 years), with proportionally fewer females (47.9% vs. 60.1%), and proportionally more of the nifurtimox group than the untreated group had clinical signs and symptoms of Chagas disease at diagnosis (28.9% vs. 14.0%). Median maximum daily dose of nifurtimox was 8.0 mg/kg/day (interquartile range [IQR]: 8.0-9.0) and median treatment duration was 44 days (IQR: 1-90). Median time to event was 2.1 years (IQR: 1.0-4.5) for nifurtimox-treated and 2.4 years (IQR: 1.0-4.2) for untreated patients. Accounting for potential confounders, the estimated hazard ratio (95% confidence interval) for negative seroconversion was 2.22 (1.61-3.07) favoring nifurtimox. Variable treatment regimens and follow-up duration, and an uncommonly high rate of spontaneous negative seroconversion, complicate interpretation of this epidemiological study, but with the longest follow-up and largest cohort analyzed to date it lends weight to the benefit of nifurtimox in adults with chronic Chagas disease. Trial registration: The study protocol was registered at ClinicalTrials.gov: NCT03784391.

Prospective, historically controlled study to evaluate the efficacy and safety of a new paediatric formulation of nifurtimox in children aged 0 to 17 years with Chagas disease one year after treatment (CHICO).

Nifurtimox is a recommended treatment for Chagas disease, but data from treated children are limited. We investigated the efficacy, safety and tolerability of nifurtimox administered as divisible, dispersible 30 mg and 120 mg tablets in children with Chagas disease. In this blinded, controlled study conducted January 2016-July 2018, 330 patients aged <18 years from 25 medical centres across three South American countries were randomised 2:1 to nifurtimox 10-20 mg/kg/day (aged <12 years) or 8-10 mg/kg/day (aged ≥12 years) for 60 days (n = 219), or for 30 days plus placebo for 30 days (n = 111) (ClinicalTrials.gov NCT02625974). The primary outcome was anti-Trypanosoma cruzi serological response (negative seroconversion or seroreduction ≥20% in mean optical density from baseline determined by two conventional enzyme-linked immunosorbent assays) at 12 months in the 60-day treatment group versus historical placebo controls. Nifurtimox for 60 days achieved negative seroconversion (n = 10) and seroreduction (n = 62) in 72 patients (serological response 32.9%; 95% confidence interval [CI] 26.4%, 39.3%, of all treated patients), confirming superiority relative to the upper 95% CI of 16% for controls. In patients aged <8 months, 10/12 treated for 60 days (83.3%) and 5/7 treated for 30 days (71.4%) achieved negative seroconversion. Overall serological response was lower for 30-day than for 60-day nifurtimox (between-treatment difference 14.0% [95% CI 3.7%, 24.2%]). The frequency of T. cruzi-positive quantitative polymerase chain reactions decreased substantially from baseline levels (60-day regimen 53.4% versus 1.4%; 30-day regimen 51.4% versus 4.5%). Study drug-related treatment-emergent adverse events (TEAEs), which were observed in 62 patients (28.3%) treated for 60 days and 29 patients (26.1%) treated for 30 days, were generally mild or moderate and resolved without sequelae; 4.2% of all TEAEs led to nifurtimox discontinuation. Age- and weight-adjusted nifurtimox for 60 days achieved a serological response at 12 months post-treatment that was superior to historical placebo, was well tolerated and had a favourable safety profile in children with Chagas disease. Although, at 1 year serological follow-up, efficacy of the shorter nifurtimox treatment was not comparable to the 60-day treatment regimen for the overall study population, further long-term follow-up of the patients will provide important information about the progress of serological conversion in children treated with nifurtimox, as well as the potential efficacy difference between the two regimens over time.

Chaperone activity of large-size subunit catalases.

Large-size subunit catalases (LSCs) have a C-terminal domain that is structurally similar to DJ-1 and Hsp31 proteins, which have well documented molecular chaperone activity. Like chaperones, LSCs are abundant proteins that are induced under stress conditions and during cell differentiation in different microorganisms. Here we document that the C-terminal domain of LSCs assist other proteins to preserve their active conformation. Heat, urea, or H2O2 denaturation of alcohol dehydrogenase was prevented by LSCs or the C-terminal domain of Catalase-3 (TDC3); in contrast, small-size subunit catalases (SSCs) or LSCs without the C-terminal domain (C3ΔTD or C63) did not have this effect. Similar results were obtained if the alcohol dehydrogenase was previously denatured by heat and then the different catalases or truncated enzymes were added. The TDC3 also protected both the C3ΔTD and the bovine liver catalase from heat denaturation. The chaperone activity of CAT-3 or the TDC3 increased survival of E. coli under different stress conditions whereas the C3ΔTD did not. It is concluded that the C-terminal domain of LSCs has a chaperone activity that is instrumental for cellular resistance to stress conditions, such as oxidative stress that leads to cell differentiation in filamentous fungi.

Opioid modulation of cochlear auditory responses in the rat inner ear.

The auditory system has an extensive efferent innervation, which contributes to processes of control and regulation of the afferent input. The expression of receptors to various neurotransmitters and neuropeptides in the inner ear has been described, among which endogenous opioid receptors are found. The role of opioid receptors in the cochlea is not yet fully defined, it has been reported that opioid agonists and antagonists modulate the response to auditory stimuli and in clinical practice, multiple cases have been reported in which the consumption of opioid derivatives induce sensorineural hearing loss. In this work, we evaluated the effects of acute treatment with morphine, fentanyl, tramadol, and naloxone, in the auditory brain stem potentials (ABR), the compound action potential (CAP), and distortion products otacoustic emissions (DPOAE), across a wide range of stimulus frequencies and amplitudes. Adult Long-Evans rats of the strain CII/ZV weighing 180-220 g were used. For the ABR recording drugs were administered intraperitoneally or intravenously. For the CAP and DPOAE drugs were applied by direct perfusion in the middle ear. The opioid agonists produced a consistent increase in the amplitude of the PI component of the ABR and of the N1-P1 amplitude of the CAP. Naloxone produced no significant changes in the ABR and a reduction of the CAP N1-P1 amplitude. Also, opioid agonists induced a decrease in the amplitude of the DPOAE. These results show that the opioid receptor activation modulates both the afferent response at both the afferent response to acoustic stimuli, and also at the cochlear mechanics as revealed by DPOAE changes. These results present a significant step in understanding how opioid modulation of auditory responses may contribute to the auditory processing and to sensorineural hearing loss produced by opioids.

Gender equity in physical education: The use of language / Igualdade de gênero em educação física: o uso da linguagem / La equidad de género en la educación física: el uso de lenguaje

This study analyzed Spanish teachers' behavior and the transmission of gender stereotypes. We observed 48 physical education lessons given by four Spanish teachers (two men and two women). Descriptive codes, which were generated iteratively, were clustered, categorized, integrated, recoded, and re-categorized. They allowed us to identify four major themes related to the transmission of gender stereotypes of teachers: male generics, stereotyped expressions, nominative attention, and priority order. We used a coding sheet as well as audio and video recordings to register the categories. The Kruskal-Wallis test produced significance levels lower than .05, resulting in the rejection of the null hypothesis. Sexist behavior was found in the male generics, nominative attention, and priority order. However, we found no difference in stereotyped expressions...

"Igualdade de gênero em educação física: O uso da linguagem." O objetivo do estudo foi analisar o comportamento dos professores espanhóis na transmissão de estereótipos de gênero para os seus alunos. Observou-se 48 aulas de educação física dadas por quatro professores espanhóis (dois homens e duas mulheres). Os códigos descritivos, que foram gerados de forma iterativa, foram agrupados, categorizados, integrados, recodificados e reclassificados. Identificou-se quatro variáveis relacionando o professorado e a sua transmissão dos estereótipos de gênero: masculinos genéricos, expressões estereotipadas, atenção nominativa e ordem de prioridade. Utilizou-se uma folha de codificação, bem como gravações de áudio e vídeo para os registros. Níveis de significância no teste de Kruskal-Wallis inferiores 0,05 permitiram rejeitar a hipótese nula. Comportamentos sexistas foram encontrados no uso de masculino genérico, a atenção nominativa e ordem de prioridade. No entanto, não foi encontrada diferenças nas expressões estereotipadas...

"La equidad de género en la educación física: El uso del lenguaje." El objetivo de este estudio es analizar el comportamiento de los profesores españoles en la transmisión de estereotipos de género hacia su alumnado. Para ello, se han observado 48 clases de educación física impartidas por cuatro profesores españoles (dos hombres y dos mujeres). Los códigos descriptivos que se generaron de forma interactiva, fueron agrupados, categorizados, integrados, recodificados y recategorizados. Se identificaron cuatro variables relacionadas con el profesorado y su transmisión de estereotipos de género: masculino genérico, expresiones estereotipadas, atención nominativa y orden de prelación. Para el registro de las categorías se utilizó una hoja de observación, junto con grabaciones de audio y vídeo. Niveles de significación en el test de Kruskal-Wallis inferiores a ,05 permitieron rechazar la hipótesis nula. Se encontraron comportamientos sexistas en el uso del masculino genérico, la atención nominativa y el orden de prelación. Sin embargo, no se encontraron diferencias en las expresiones estereotipadas...

Actitud de las enfermeras sobre el Método Madre Canguro en el Servicio de UCIN de la Clínica Maison de Santé-2012 / Attitude of the nurses about Kangaroo-Mother Care Method in the NICU Service at the Clinic Maison de Sante 2012

El Objetivo fue determinar la actitud de las enfermeras sobre el Método Madre Canguro en el servicio de UCIN de la Clínica Maison de Santé - 2012. Material y Método: El estudio fue de nivel aplicativo, tipo cuantitativo, método descriptivo de corte transversal. La población estuvo conformada por 20 enfermeras. La técnica fue la encuesta y el instrumento la Escala modificada de Likert, aplicado previo consentimiento informado. Resultados: Del 100 por ciento (20), 65 por ciento (13) tienen actitud de indiferencia, 15 por ciento (3) aceptación y 20 por ciento (4) rechazo. En la dimensión lactancia materna el 85 por ciento (17) tienen actitud de indiferencia, 5 por ciento (1) de aceptación y 10 por ciento (2) rechazo. En la dimensión de vínculo afectivo 60 por ciento (12) actitud de indiferencia, y 20 por ciento (4) actitud de rechazo; y en la dimensión de termorregulación del prematuro el 50 por ciento (10) actitud de indiferencia, 25 por ciento (05) actitud de aceptación. Conclusiones: La actitud de las enfermeras sobre el Método Madre Canguro en la mayoría fue de indiferencia y rechazo, con un mínimo porcentaje de aceptación. Según las dimensiones la mayoría de las enfermeras presenta una actitud de indiferencia frente a la dimensión de Lactancia Materna Exclusiva y un mínimo porcentaje de aceptación en la Termorregulación del Prematuro. En relación a los ítems la mayoría de las enfermeras presenta una actitud de indiferencia frente al beneficio inmunológico y cognitivo, relación madre e hijo, inmadurez térmica.

The objective was to determine the attitude of the nurses about KMC in the NICU service of the Clinic Maison de Sante - 2012. Material and Methods: The study was level application, quantitative, descriptive cross-sectional method. The sample consisted of 20 nurses. The technique was the survey instrument and modified Likert Scale, applied prior informed consent. Results: 100 per cent (20), 65 per cent (13) have indifference, 15 per cent (3) accepting attitude and 20 per cent (4) of rejection. Breastfeeding in the size 85 per cent (17) have indifference, 5 per cent (1) acceptance. In the dimension of bonding 60 per cent (12) attitude of indifference, and 20 per cent (4) attitude of rejection, and the dimension of thermoregulation of prematurity 50 per cent (10) attitude of indifference, 25 per cent (05) attitude acceptance. Conclusions: The attitude of the nurses on the Kangaroo Mother Method in most was one of indifference and rejection, with a minimum percentage of acceptances. According to the dimensions the majority of the nurses present an attitude of indifference to the dimension of exclusive breastfeeding and a minimum percentage of acceptances in the thermoregulation of the premature. In relation to the items the majority of the nurses presented attitude of indifference to the immune and cognitive benefit, mother and son relationship, immaturity thermal.

Costs of breast cancer treatment prior to the introduction of immune-based therapy in Mexico.

BACKGROUND: the aim was to perform a cost analysis on direct data of integral medical care provided to BC patients prior to introduction of immunotherapy. METHODS: a total of 633 patients were studied. Direct costs calculations were performed on individual patient data in a subset of 309 randomly selected patients, extrapolating calculations to the universe of 633 patients. Information was obtained for each management process (diagnosis and staging, cancer management, symptoms control, palliative care and follow-up). Costs are expressed in 2008 US dollars (USD) and adjusted to a 3 % discount rate. RESULTS: the clinical stage distribution in the 633 patients was 41, 191, 240 and 58 patients for clinical stages I to IV, respectively; with 103 patients referred from other institutions without staging. The annual costs of care per patient was, in clinical stage I: 6,219,94 USD; stage II: 7,498.04 USD; stage III: 9,610.31 USD; stage IV: 9,917.82 USD; and in patients without staging: 7,504.41 USD. The exact total integral cost according to the universe of BC patients (n = 633) by 2004 was 5,341,805.37 USD. CONCLUSIONS: before the introduction of immune-based therapy costs of care for Mexican women with BC increased with advanced stages do to a significant proportion of patients were diagnosed late.

Luxofractura anterior del hombro derecho con luxación posterior del codo: presentación de un caso / Previous dislocation and fracture of the right shoulder with posterior dislocation of the elbow: a case report

La luxofractura de hombro y la luxación de codo están descritas, pero son poco frecuente que ocurran en el mismo hemicuerpo. Caso clínico: paciente de 45 años, masculino, de piel blanca, que acudió al servicio de Traumatología del Hospital Universitario Martín Chang Puga de Nuevitas por presentar luxofractura de hombro y luxación posterior de codo en un mismo hemicuerpo, a causa de un accidente automovilístico. Se realizó reducción manual de urgencia de ambas luxaciones y control radiológico, con resultados satisfactorios. Se inmovilizó el codo con una férula posterior de yeso para el miembro superior y vendaje de Velpeau para el hombro. Se retiró inmovilización del codo a las tres semanas y la del hombro a las seis semanas. Posteriormente se comenzó con la rehabilitación, donde se obtuvo magníficos resultados con la función de ambas articulaciones y del miembro superior en general (AU)

Dislocation and fracture of shoulder and dislocation of elbow are described but is very unusual when they are concomitant and on the same side of the body.Clinical case: a case of a 45-year-old, male patient who was treated in the service of Traumatology at the University Hospital Martín Chang Puga from Nuevitas, presenting dislocation and fracture of the right shoulder and dislocation of the elbow on the same side of the body due to a car accident. Urgent manual reduction of both dislocations and radiological control, with satisfactory results took place. The elbow was immobilized with a backboard plaster splint and a Velpeau´s bandage for the shoulder. The immobilization devise of the elbow was removed after three weeks and that of the shoulder after six. Subsequently began rehabilitation, and great results were obtained with both joints and the upper limb functions (AU)

Luxofractura anterior del hombro derecho con luxación posterior del codo: presentación de un caso / Previous dislocation and fracture of the right shoulder with posterior dislocation of the elbow: a case report

La luxofractura de hombro y la luxación de codo están descritas, pero son poco frecuente que ocurran en el mismo hemicuerpo. Caso clínico: paciente de 45 años, masculino, de piel blanca, que acudió al servicio de Traumatología del Hospital Universitario Martín Chang Puga de Nuevitas por presentar luxofractura de hombro y luxación posterior de codo en un mismo hemicuerpo, a causa de un accidente automovilístico. Se realizó reducción manual de urgencia de ambas luxaciones y control radiológico, con resultados satisfactorios. Se inmovilizó el codo con una férula posterior de yeso para el miembro superior y vendaje de Velpeau para el hombro. Se retiró inmovilización del codo a las tres semanas y la del hombro a las seis semanas. Posteriormente se comenzó con la rehabilitación, donde se obtuvo magníficos resultados con la función de ambas articulaciones y del miembro superior en general

Dislocation and fracture of shoulder and dislocation of elbow are described but is very unusual when they are concomitant and on the same side of the body.Clinical case: a case of a 45-year-old, male patient who was treated in the service of Traumatology at the University Hospital Martín Chang Puga from Nuevitas, presenting dislocation and fracture of the right shoulder and dislocation of the elbow on the same side of the body due to a car accident. Urgent manual reduction of both dislocations and radiological control, with satisfactory results took place. The elbow was immobilized with a backboard plaster splint and a Velpeau´s bandage for the shoulder. The immobilization devise of the elbow was removed after three weeks and that of the shoulder after six. Subsequently began rehabilitation, and great results were obtained with both joints and the upper limb functions

Evidence of an intracellular angiotensin-generating system and non-AT1, non-AT2 binding site in a human pancreatic cell line.

OBJECTIVES: To assess the presence of a local angiotensin-generating systems (LAGS) and its participation in tumor growth in the human pancreatic cancer derived cell line Capan-1. METHODS: Capan-1 cells were cultured in Dulbecco modified Eagle medium, and angiotensin I was assayed by radioimmunoassay and angiotensin II and vascular endothelial growth factor were assayed by enzyme-linked immunosorbent assay in the supernatant. Immunohistochemistry and reverse transcription-polymerase chain reaction were performed for the expression of AT1 and AT2 receptors. Angiotensin II binding assays and blockade were studied. RESULTS: High levels of both angiotensins I and II were found in Capan-1 cells, although neither angiotensin I nor angiotensin II was detected in the cell culture supernatant. Reverse transcription-polymerase chain reaction and immunocytochemistry revealed that Capan-1 cells do not express AT1 and AT2 receptors; however, specific binding to the cell membrane was identified for angiotensin II. Neither exogenous angiotensin II nor Dup753 (specific AT1 receptor blocker) affected Capan-1 cells' proliferation or vascular endothelial growth factor secretion. CONCLUSIONS: Detection of both angiotensin I and angiotensin II along with specific binding of angiotensin II in Capan-1 cells provides evidence of the existence of a LAGS that operates in an intracrine manner. Intracellular angiotensin II may play a role in the aggressiveness of pancreatic cancer and is a possible target for therapeutic agents.

Detecting abnormal vehicular dynamics at intersections based on an unsupervised learning approach and a stochastic model.

This investigation demonstrates an unsupervised approach for modeling traffic flow and detecting abnormal vehicle behaviors at intersections. In the first stage, the approach reveals and records the different states of the system. These states are the result of coding and grouping the historical motion of vehicles as long binary strings. In the second stage, using sequences of the recorded states, a stochastic graph model based on a Markovian approach is built. A behavior is labeled abnormal when current motion pattern cannot be recognized as any state of the system or a particular sequence of states cannot be parsed with the stochastic model. The approach is tested with several sequences of images acquired from a vehicular intersection where the traffic flow and duration used in connection with the traffic lights are continuously changed throughout the day. Finally, the low complexity and the flexibility of the approach make it reliable for use in real time systems.

Structured interruptions of highly active antiretroviral therapy in cycles of 4 weeks off/12 weeks on therapy in children having a chronically undetectable viral load cause progressively smaller viral rebounds.

OBJECTIVES: To evaluate the viral, immune and clinical impact of a structured treatment interruption (STI) program of highly active antiretroviral therapy (HAART) in three cycles of 4 weeks off/12 weeks on therapy in a cohort of children with HIV infection under chronic viral control. METHODS: Using a single-group time series experimentation design and following informed consent, the HAART of children with HIV and a chronically undetectable viral load (VL) was discontinued for 4 weeks and then restarted and continued for 12 weeks for a total of three cycles. The VL, CD4+/CD8+ lymphocytes, and clinical status were evaluated at the end of each STI and at 6 and 12 weeks after HAART was resumed. RESULTS: Four children with a median age of 10.3 years (range 6.5-11.2 years) were included in the study. Their clinical immune categories were: A1 (n=2), A2 (n=1), and B3 (n=1). Treatment of all four patients was with zidovudine (AZT)+lamivudine (3TC)+ritonavir (RTV). At the end of the first STI, VL was a median 214000 copies/ml (range 27400-616000), corresponding to 5.3 log(10) (range 4.4-5.8). At the end of the second STI, VL was a median 72400 copies/ml (range 17800-126000) or 4.7 log(10) (range 4.2-5.1), which corresponds to a rebound 0.6 log(10) lower than the first. At the end of the third STI, VL was a median 28200 copies/ml (range 5370-140000) or 4.45 log(10) (range 3.7-5.1), a rebound 0.85 log(10) lower than the first. All rebounds were followed by a decrease in the VL to undetectable levels during the treatment periods. CD8+ T lymphocyte counts increased during viral rebounds and an initial decrease in CD4+ T lymphocyte counts was followed by a tendency to increase even exceeding CD8+ T cell counts. Only one event of transitory severe immunosuppression occurred. There were no symptoms related to the HIV infection. CONCLUSIONS: The STI of HAART in cycles of 4 weeks off/12 weeks on therapy in children with chronically undetectable VL can cause progressively lower viral rebounds followed by a decrease to undetectable levels, with a low risk of severe immunosuppression and without the occurrence of symptoms related to HIV.

Intraepithelial gammadelta+ lymphocytes: a comparative study between celiac disease, small intestinal bacterial overgrowth, and irritable bowel syndrome.

BACKGROUND: Intraepithelial lymphocytes (IELs) phenotyping has emerged as a useful test in intestinal pathology. In celiac disease (CD), a permanent and marked increase of gammadelta+ IELs has been described. However, there is a lack of knowledge about this peculiar IELs population in other intestinal pathologies. AIM: To analyze the percentage of IELs, specifically gammadelta+ IELs subset, present in duodenal mucosa biopsies from patients with CD and compare it with those obtained from patients with small intestinal bacterial overgrowth (SIBO) or irritable bowel syndrome (IBS). METHODS: Twelve patients with untreated CD, 8 patients with SIBO, and 10 patients with diarrhea-predominant IBS were evaluated. All subjects underwent upper endoscopy for mucosal biopsy and jejunal aspirate. From 2 small bowel biopsies, intraepithelial cells were isolated and labeled with the following monoclonal antibodies CD103-PE (phycoerythrin), CD3-FITC (fluoresecein isothio-cynate), CD-7R-PE, CD45RO-APC (allophycocyanin), and TcR gammadelta-FITC. Flow cytometry analysis was performed on a standard FACScan. Total and IELs subset counts were expressed as percentage. RESULTS: Mean total IELs percentage was 16.7+/-6% in IBS, 25.4+/-17% in SIBO, and 26+/-13% in CD patients (P=0.2). CD and SIBO patients, had significantly higher percentages of gammadelta+ IELs (15.7+/-13% and 14.6+/-8%) than IBS subjects (4.1+/-2.5%, P<0.05). There was no difference between CD and SIBO (P=0.6). CONCLUSIONS: An increased density of gammadelta+ IELs is typical, but not specific for CD. A similar increase was observed in subjects with SIBO. Our findings suggest that this unique T-cell population might have a key role against intestinal bacterial infections.

A proof-of-principle study of epigenetic therapy added to neoadjuvant doxorubicin cyclophosphamide for locally advanced breast cancer.

BACKGROUND: Aberrant DNA methylation and histone deacetylation participate in cancer development and progression; hence, their reversal by inhibitors of DNA methylation and histone deacetylases (HDACs) is at present undergoing clinical testing in cancer therapy. As epigenetic alterations are common to breast cancer, in this proof-of-concept study demethylating hydralazine, plus the HDAC inhibitor magnesium valproate, were added to neoadjuvant doxorubicin and cyclophosphamide in locally advanced breast cancer to assess their safety and biological efficacy. METHODOLOGY: This was a single-arm interventional trial on breast cancer patients (ClinicalTrials.gov Identifier: NCT00395655). After signing informed consent, patients were typed for acetylator phenotype and then treated with hydralazine at 182 mg for rapid-, or 83 mg for slow-acetylators, and magnesium valproate at 30 mg/kg, starting from day -7 until chemotherapy ended, the latter consisting of four cycles of doxorubicin 60 mg/m2 and cyclophosphamide 600 mg/m2 every 21 days. Core-needle biopsies were taken from primary breast tumors at diagnosis and at day 8 of treatment with hydralazine and valproate. MAIN FINDINGS: 16 patients were included and received treatment as planned. All were evaluated for clinical response and toxicity and 15 for pathological response. Treatment was well-tolerated. The most common toxicity was drowsiness grades 1-2. Five (31%) patients had clinical CR and eight (50%) PR for an ORR of 81%. No patient progressed. One of 15 operated patients (6.6%) had pathological CR and 70% had residual disease <3 cm. There was a statistically significant decrease in global 5mC content and HDAC activity. Hydralazine and magnesium valproate up- and down-regulated at least 3-fold, 1,091 and 89 genes, respectively. CONCLUSIONS: Hydralazine and magnesium valproate produce DNA demethylation, HDAC inhibition, and gene reactivation in primary tumors. Doxorubicin and cyclophosphamide treatment is safe, well-tolerated, and appears to increase the efficacy of chemotherapy. A randomized phase III study is ongoing to support the efficacy of so-called epigenetic or transcriptional cancer therapy.

Synthesis and pharmacological evaluation of new progesterone esters as 5alpha-reductase inhibitors.

In this study we report the synthesis and pharmacological evaluation of four new progesterone derivatives; 17alpha-hydroxy-16beta-methylpregna-4,6-diene-3,20-dione 12, 17alpha-cyclopropylcarbonyloxy-16beta-methylpregna-4,6-diene-3,20-dione 13, 17alpha-cyclobutylcarbonyloxy-16beta-methylpregna-4,6-diene-3,20-dione 14, 17alpha-acetoxy-16beta-methylpregna-4,6-diene-3,20-dione 15 and the pregnatriene compound 17alpha-cyclobutylcarbonyloxy-16beta-methylpregna-1,4,6-triene-3,20-dione 16. The pharmacological effect of these compounds was determined in vivo as well as in vitro. The evaluation in vivo was carried out on gonadectomized male hamsters that were injected subcutaneously daily with testosterone (T) and/or finasteride, or with the novel compounds. At the end of the treatments the animals were sacrificed and the prostates were weighed. It was observed that when testosterone (T) and finasteride or compounds 12-16 were injected together, the weight of the prostate decreased significantly as compared to that of the testosterone-treated animals. The 5alpha-reductase inhibitory activity was evaluated in vitro using human prostate homogenates. These experiments showed the following IC50 values: compound 12 (alcohol at C-17) 1.2 x 10(-6) M, 13 (cyclopropyl substituent at C-17) 7.9 x 10(-10) M, 14 (cyclobutyl substituent) 3.2 x 10(-8) M, 15 (acetoxy substituent) 6.3 x 10(-11) M and 16 (cyclobutyl substituent) 3.9 x 10(-6) M. It is evident from these data that when the size of the substituent at C-17 is decreased, the 5alpha-reductase inhibitory activity increases. Apparently, in this biological model, the 5alpha-reductase inhibitory activity depends upon the steric effect of the substituent at C-17. However, the free alcohol 12 showed much lower 5alpha-reductase inhibitory activity.

Computer-based education for patients with cancer at Latino border hospital.

There is a paucity of relevant educational material for Latino patients with cancer. Education interventions used for non-Latinos must be assessed for relevancy for Latino patients. This study examined the use and usefulness of computer-based education interventions among Latinos. It was conducted using the CancerHelp software survey in a Texas institution on the US-Mexico border. Nonparametric test statistics were used to test the hypothesized relationships and the usefulness of CancerHelp software. The following statistic hypotheses were not rejected: (1) CancerHelp use is independent of age group, (2) use of CancerHelp is equally distributed among patients and family members, (3) among Latinos, CancerHelp use is independent of education, and (4) CancerHelp usefulness for patients with cancer is independent of ethnicity.