TGF-ß1 decreases CHOP expression and prevents cardiac fibroblast apoptosis induced by endoplasmic reticulum stress.

Transforming growth factor-beta 1 (TGF-ß1) is a cytokine with marked pro-fibrotic action on cardiac fibroblasts (CF). TGF-ß1 induces CF-to-cardiac myofibroblast (CMF) differentiation, defined by an increase in α-smooth muscle cells (α-SMA), collagen secretion and it has a cytoprotective effect against stimuli that induce apoptosis. In the Endoplasmic Reticulum (ER) lumen, misfolded protein accumulation triggers ER stress and induces apoptosis, and this process plays a critical role in cell death mediated by Ischemia/Reperfusion (I/R) injury and by ER stress inducers, such as Tunicamycin (Tn). Here, we studied the regulation of CHOP, a proapoptotic ER-stress-related transcription factor in CF under simulated I/R (sI/R) or exposed to Tn. Even though TGF-ß1 has been shown to participate in ER stress, its regulatory effect on CF apoptosis and ER stress-induced by sI/R or TN has not been evaluated yet. CF from neonatal rats were exposed to sI/R, and cell death was evaluated by cell count and apoptosis by flow cytometry. ER stress was assessed by western blot against CHOP. Our results evidenced that sI/R (8/24) h or Tn triggers CF apoptosis and an increase in CHOP protein levels. TGF-ß1 pre-treatment partially prevented apoptosis induced by sI/R or Tn. Furthermore, TGF-ß1 pre-treatment completely prevented CHOP increase by sI/R or Tn. Additionally, we found a decrease in α-SMA expression induced by sI/R and in collagen secretion induced by Tn, which were not prevented by TGF-ß1 treatment. In conclusion, TGF-ß1 partially protects CF apoptosis induced by sI/R or Tn, through a mechanism that would involve ER stress.

Osteomielitis vertebral por Bartonella henselae: a propósito de un caso / Bartonella henselae vertebral osteomyelitis: report of a case

Resumen: La enfermedad por arañazo de gato (EAG) causada por Bartonella henselae tiene prevalencia e incidencia desconocida en la población pediátrica chilena. La presentación como linfoadenopatía regional es la más frecuente, y sus formas atípicas constituyen un reto diagnóstico. Objetivo: Comunicar un caso de EAG con osteomielitis vertebral y entregar posibles lineamientos respecto al tratamiento. Cuadro clínico: Escolar de 8 años con síndrome febril prolongado y dolor dorsal con rigidez cervical. En estudio destaca IgG para Bartonella henselae positiva, ecotomografía abdominal con microabscesos esplénicos y resonancia magnética con lesiones vertebrales sugerentes de osteomielitis. Discusión: El diagnóstico de las formas atípicas requiere de un alto índice de sospecha, como en este caso, en que el paciente con síndrome febril manifiesta sintomatología musculoesquelética, lo que nos lleva a estudiar posibles complicaciones de la enfermedad. El conocimiento actual del tratamiento de EAG atípica o complicada se deriva de la observación de estudios de casos y no de ensayos aleatorios; se sugiere analizar de manera individual la terapia antimicrobiana y con ayuda de especialistas. Conclusión: Se discute y enfatiza la importancia de la alta sospecha clínica y se exponen algunas alternativas de tratamiento en función de la evidencia de la literatura actual.

Abstract: Cat scratch disease (CSD) is caused by Bartonella henselae, with unknown prevalence and incidence in the Chilean paediatric population. Regional lymphadenopathy is the most common presentation, while atypical forms constitute a diagnostic challenge. Objective: To report a case of CSD with osteomyelitis and present guidelines regarding treatment. Clinical case: An eight year-old patient, with prolonged febrile illness, back pain and neck stiffness. Laboratory studies highlight positive IgG for Bartonella henselae. The abdominal ultrasound showed splenic micro-abscesses, and the MRI showing vertebral lesions suggestive of osteomyelitis. Discussion: The diagnosis of atypical forms requires a high rate of suspicion, as in this case, in which the patient manifested the musculoskeletal symptoms simultaneously with the febrile syndrome, which led us to study possible complications of the disease. Current knowledge of the treatment of atypical or complicated CSD is derived from the observation of case studies, rather than randomized trials. It is suggested that antibiotic therapy is analysed individually, with the help of a specialist. Conclusion: The importance of high clinical suspicion are emphasised and discussed, as well presenting some treatment options based on the evidence from the current literature.

Mutation analysis of Swedish haemophilia B families - high frequency of unique mutations.

INTRODUCTION: Haemophilia B is caused by a heterogeneous spectrum of mutations. Mutation characterization is important in genetic counselling, prenatal diagnosis and to predict risk of inhibitor development. AIMS: To study the mutation spectrum, frequency of unique recurrent mutations, genotype-phenotype association and inhibitor development in a population-based study of the complete Swedish haemophilia B population. METHODS: The study included, facilitated by centralized DNA diagnostics, the complete registered Swedish haemophilia B population (113 families: 47 severe, 22 moderate and 44 mild), each represented by a single patient. Mutation characterization was performed by conventional sequencing of all exons and haplotyping by genotyping of single nucleotide variants and microsatellites. RESULTS: A mutation was found in every family: eight had large deletions, three had small deletions (<10 base pair) and 102 had single base pair substitutions (69 missense, 26 nonsense, four splice site and three promoter). Ten novel mutations were found and were predicted to be deleterious. Sixteen mutations (one total gene deletion, 14 substitutions and one acceptor splice site) were present in more than one family. Of the single nucleotide mutations (37/102), 36% arose at CpG sites. Haplotyping of families with identical mutations and present analyses showed that the frequency of unique mutations was at least 65%. Inhibitors developed in 9/47 (19%) patients with severe haemophilia B. CONCLUSION: The spectrum of haemophilia B mutations reveals at least 65% of the families carry a unique mutation, but with more inhibitor patients than reported internationally, probably as a result of many 'null' mutations.

[Bartonella henselae vertebral osteomyelitis: report of a case]. / Osteomielitis vertebral por Bartonella henselae: a propósito de un caso.

UNLABELLED: Cat scratch disease (CSD) is caused by Bartonella henselae, with unknown prevalence and incidence in the Chilean paediatric population. Regional lymphadenopathy is the most common presentation, while atypical forms constitute a diagnostic challenge. OBJECTIVE: To report a case of CSD with osteomyelitis and present guidelines regarding treatment. CLINICAL CASE: An eight year-old patient, with prolonged febrile illness, back pain and neck stiffness. Laboratory studies highlight positive IgG for Bartonella henselae. The abdominal ultrasound showed splenic micro-abscesses, and the MRI showing vertebral lesions suggestive of osteomyelitis. DISCUSSION: The diagnosis of atypical forms requires a high rate of suspicion, as in this case, in which the patient manifested the musculoskeletal symptoms simultaneously with the febrile syndrome, which led us to study possible complications of the disease. Current knowledge of the treatment of atypical or complicated CSD is derived from the observation of case studies, rather than randomized trials. It is suggested that antibiotic therapy is analysed individually, with the help of a specialist. CONCLUSION: The importance of high clinical suspicion are emphasised and discussed, as well presenting some treatment options based on the evidence from the current literature.

Origin of mutation in sporadic cases of severe haemophilia A in Sweden.

UNLABELLED: Many newly diagnosed Swedish severe haemophilia A (HA) patients are sporadic cases. Some genotypically non-carrier mothers have gone on to have two descendants with the same mutation, presumably because of mosaicism. AIMS: To define the origin of mutation in sporadic cases of HA, reveal possible sex-specific differences in mutagenesis and identify potential mosaics among non-carrier mothers. METHOD: Sanger sequencing characterized the mutations and microsatellite haplotyping determined the origin of the X chromosome carrying the mutation in 3 generations of 45 families with sporadic severe HA. Droplet digital polymerase chain reaction (ddPCR) was used in five cases to reveal that mosaicism mutations are not found on conventional DNA sequencing. RESULTS: In 23 out of 45 families, the mother carried the mutation and in 5 out of 28 families, the grandmother was also a carrier. The X chromosome was of grandpaternal origin in 17 out of 23 cases. One of five tested mothers was a mosaic with a mutation frequency of 7%. CONCLUSION: In 40 out of 45 families, the sporadic case resulted from a mutation in the last two generations. In 82% (23/28), the carrier mothers had a de novo mutation where the X chromosome was of paternal origin in 74% (17/23). ddPCR is a potentially powerful and promising analysis for mosaicism in HA.

4-Phenylbutyric acid prevent cytotoxicity induced by thapsigargin in rat cardiac fibroblast.

Cardiac fibroblast (CF) survival is important for the maintenance of the extracellular matrix homeostasis in the heart; providing a functional support to cardiomyocytes necessary for the correct myocardial function. Endoplasmic reticulum (ER) stress causes cellular dysfunction and cell death by apoptosis; and thapsigargin is a well-known ER stress inducer. On the other hand, the chemical chaperone, 4-phenylbutyric acid (4-PBA) had showed to prevent ER stress; however, in cardiac fibroblast both the ER stress induced by thapsigargin and prevention by 4-PBA, have not been studied in detail. Neonate rat CF were treated with thapsigargin in presence or absence of 4-PBA, and cell viability was evaluated by trypan blue exclusion and apoptosis by flow cytometry; whereas CHOP, BIP, PDI, ATF4 and procollagen protein levels were assessed by western blot. In CF, thapsigargin triggered the unfolded protein response detected by early increases in ATF4, CHOP, PDI and BIP protein levels as well as, the accumulation of intracellular procollagen. Thapsigargin also stimulated CF death in a time and concentration-dependent manner. ER stress, CF death and apoptosis induced by thapsigargin were prevented by 4-PBA. Conclusion our data suggest that 4-PBA prevent ER stress, intracellular procollagen accumulation, CF death and apoptosis induced by thapsigargin.

Landscape heterogeneity shapes host-parasite interactions and results in apparent plant-virus codivergence.

Knowledge on how landscape heterogeneity shapes host-parasite interactions is central to understand the emergence, dynamics and evolution of infectious diseases. However, this is an underexplored subject, particularly for plant-virus systems. Here, we analyse how landscape heterogeneity influences the prevalence, spatial genetic structure, and temporal dynamics of Pepper golden mosaic and Pepper huasteco yellow vein begomoviruses infecting populations of the wild pepper Capsicum annuum glabriusculum (chiltepin) in Mexico. Environmental heterogeneity occurred at different nested spatial scales (host populations within biogeographical provinces), with levels of human management varying among host population within a province. Results indicate that landscape heterogeneity affects the epidemiology and genetic structure of chiltepin-infecting begomoviruses in a scale-specific manner, probably related to conditions favouring the viruses' whitefly vector and its dispersion. Increased levels of human management of the host populations were associated with higher virus prevalence and erased the spatial genetic structure of the virus populations. Also, environmental heterogeneity similarly shaped the spatial genetic structures of host and viruses. This resulted in the congruence between host and virus phylogenies, which does not seem to be due to host-virus co-evolution. Thus, results provide evidence of the key role of landscape heterogeneity in determining plant-virus interactions.

Utilización de resonancia magnética funcional 3T para evaluar los efectos del metilfenidato sobre la función de atención en pacientes con déficit atencional / Using 3T functional magnetic resonance imaging to assess attention in patients with clinically diagnosed attention-deficit disorder before and after methylphenidate administration: assessment of drug effects on attentional function

Our objective was to assess the level of attention in patients with attention-deficit disorder using functional MRI by comparing test results before and after the use of methylphenidate. We studied 6 pediatric patients and 2 adults. All of them were subject to a attention test before and after methylphenidate administration. The pediatric patient population showed a positive difference, with higher levels of attention in the methylphenidate group. In the adult group, improvement in the attentional function was observed in one of them. This method allowed us to physiologically assess if improvement in attentional function occurred after methylphenidate administration. Therefore, this technique is viewed as an important tool for evaluating the usefulness of treatment for attention-deficit.hyperactivity disorder.

El objetivo es evaluar el nivel de atención utilizando resonancia funcional en pacientes con déficit atencional, comparando los resultados entre los test de atención realizados antes y después del uso de metilfenidato. Se estudiaron 6 pacientes pediátricos y 2 adultos. Cada uno realizó un test de atención antes y después de usar metilfenidato. El estudio grupal de los pacientes pediátricos mostró una diferencia positiva con mayor nivel de atención en el grupo con tratamiento con metilfenidato. En el test realizado con medicamento en los adultos se evidenció mejoría en la función de atención en uno de ellos. Este método permite medir fisiológicamente si existe mejoría en la función de atención después de la utilización de metilfenidato, constituyendo una herramienta para evaluar la utilidad del tratamiento del déficit atencional.

Implementación y validación de protocolo de estimulación por comprensión de lenguaje oral en resonancia magnética funcional: resultados preliminares / Implementation and validation of a stimulation protocol for oral language comprehension using functional magnetic resonance imaging: preliminary results

This study aimed at implementing a stimulation protocol using functional Magnetic Resonance Imaging (fMRI), in a Hospital in Valparaiso (V Region), Chile, to detect cortical areas activated in oral language comprehension. Seven healthy volunteers participated in this study. Average t-score and signal variation were 6.3 +/- 0.3 and 0.5 +/- 0.1 percent, respectively. In spite of these low values, activations were obtained in Wernicke area, middle temporal gyrus, and Heschl s gyrus. The extension of activated areas was small, 5.06 +/- 2.99 cm3, probably due to amplifer low signal-to-noise ratio (SNR), in addition to the cognitive complexity of the task, and to the ambient acoustic noise. Successful implementation of fMRI protocols of language comprehension is possible in a clinical context in Chile without any additional resources.

Nuestro objetivo fue implementar un protocolo de estimulación en resonancia magnética funcional en un Hospital de Valparaíso, V Región de Chile, para detectar las áreas corticales activadas en la comprensión del lenguaje oral. Siete voluntarios sanos participaron de este estudio. El t-score y variación de señal alcanzado fue de 6.3 0.3 y 0.5 +/- 0.1 por ciento respectivamente. A pesar de estos bajos valores, las activaciones se registraron en el área Wernicke, circunvolución temporal media y circunvolución d e Heschl. La extensión de las activaciones fue pequeña, 5.06 +/- 2.99 cm³, probablemente debido a la baja relación señal ruido del resonador (SNR), además de la complejidad cognitiva de la tarea y el ruido acústico ambiente. Nuestros resultados indican que la implementación de la RMf en comprensión del lenguaje es posible de realizar en Chile sin recursos adicionales.

Warfarina versus acenocumarol en alcanzar niveles terapéuticos en una población ambulatoria / Comparison of acenocumarol vs. warfarin in achieving oral anticoagulation therapeutic level in ambulatory patients

Introducción: La anticoagulación constituye una terapia farmacológica habitual en la práctica clínica diaria. En Chile, los ACO disponibles y utilizados son warfarina y acenocumarol, no existiendo mayores experiencias nacionales documentadas sobre el mayor beneficio de un fármaco en particular. Objetivo: Analizar y comparar la eficacia terapéutica de warfarina y acenocumarol en una población ambulatoria. Metodología: Estudio retrospectivo, longitudinal. Se analizó 188 pacientes que estuvieron en tratamiento con acenocumarol durante más de un año, y que luego fueron cambiados a warfarina. Se registró: sexo, edad, efectos adversos, diagnóstico y justificación de inicio de ACO. Se obtuvo el promedio del International Normalizad Ratio (INR) de los últimos 3 meses de tratamiento con acenocumarol. Luego, se sustituyó por warfarina, obteniendo luego de un año de tratamiento, el INR promedio de los últimos 3 meses. Los pacientes se agruparon en tres grupos: Bajo rango terapéutico (INR<2.0), en rango terapéutico (INR=2.0-3.0), sobre rango terapéutico (INR>3.0). Resultados: En los pacientes con acenocumarol, se observó 67 (35,64 por ciento) bajo rango terapéutico; 91 (48,4 por ciento) en rango terapéutico; y 30 (15,96 por ciento) sobre rango terapéutico. Luego del cambio a warfarina, 76 (40,43 por ciento) bajo rango terapéutico; 95 (50,53 por ciento) en rango terapéutico; y 17 (9,04 por ciento) sobre rango terapéutico, diferencias no significativas. Bajo el efecto de ambos fármacos no se registraron hemorragias mayores y no hubo diferencia significativa en hemorragias menores. Discusión: La eficacia terapéutica fue similar con ambos fármacos. A pesar de que con acenocumarol se obtuvo mayor porcentaje de pacientes sobre rango terapéutico, no se observaron complicaciones mayores en el periodo de seguimiento.

Background: oral anticoagulation is frequently needed in clinical practice. Warfarin and acenocumarol are available in Chile for this purpose. Locally there is no evidence favoring one over the other Aim: To compare the efficacy of warfarin and acenocumarol in an ambulatory population. Method: A retrospective study compared data on 188 patients with over 1 year of treatment with acenocumarol, before and after being switched over to treatment with warfarin. Demographic data, adverse effects, diagnosis and indication for oral anticoagulation were record. INRs obtained in the last 3 months of treatment with each agent were compared. Patients were classified in 3 groups: insufficient level (INR < 2.0), adequate level (INR 2.0- 3.0) and high level (INR > 3.0) of anticoagulation. Results: With acenocumarol, low level INR was present in 35.6 percent>, adequate INR in 48.4 percent> and high INR in 15.9 percent> of subjects. After switching to warfarin, corresponding levels in each group were 40.4 percent>, 50.3 percent> and 9 percent> (NS). There were no serious bleeding episodes in either group; minor hemorrhages occurred with similar frequency in both groups. Conclusion: There was similar clinical efficacy of oral anticoagulation with acenocumarol compared to warfarin. The slightly higher percentage of acenocumarol treated patients exhibiting a high IRN level did not result in increased risk of hemorrhage.

Erythrocyte CuZn superoxide dismutase activity is decreased in iron-deficiency anemia.

Iron and copper are essential microminerals that are intimately related. The present study was performed to determine the effect of iron-deficiency anemia (IDA) and treatment with iron on laboratory indicators of copper status. Hemoglobin, mean corpuscular volume erythrocyte Zn protoporphyrin, serum ferritin, serum copper, serum ceruloplasmin, and erythrocyte CuZn-superoxide dismutase (SOD) activity were studied in 12 adult women with IDA before and after iron treatment for 60-90 d (100 mg/d Fe, as ferric polymaltose) and in 27 women with normal iron status. Prior to treatment with iron, serum copper and ceruloplasmin were not different between the groups and treatment with iron did not affect these measures. IDA women, before and after treatment with iron, presented a 2.9- and 2-fold decrease in erythrocyte CuZn-SOD activity compared to women with normal iron status (p < 0.001). Treatment with iron increased erythrocyte CuZn-SOD activity of the IDA group; however, this change was not statistically significant. In conclusion, CuZn-SOD activity is decreased in IDA. Measurement of this enzyme activity is not useful for evaluating copper nutrition in iron-deficient subjects.

[Iron nutritional status in pregnant adolescents at the beginning of gestation]. / Situación de la nutrición de hierro en la embarazada adolescente al inicio de la gestación.

The frequency of anemia and iron nutrition deficiency was assessed in 342 low socioeconomic level pregnant teenagers at entry to prenatal care in 5 outpatient clinics from a South Orient area of Santiago Chile. According to the Center for Disease Control Criteria, 1.2% of women had iron deficiency anemia. Iron stores were insufficient (defined as a serum ferritin lower than 20 g/L) in 55% for women and depleted (serum ferritin lower than 10 g/L) in 21%. Women with more than 14 weeks of gestation had lower packed red cell volumes, hemoglobin, mean corpuscular volumes and ferritin levels than women with less than 14 of gestation. It is concluded that the prevalence of iron deficiency anemia is lower than that predicted for a highly vulnerable group but the high frequency of low iron stores should encourage the use of iron supplementation in these teenagers.

[Normal values of the hemogram and other hematological variables in subjects over 60 years old]. / Valores normales del hemograma y otras variables hematológicas en sujetos mayores de 60 años.

We studied 36 men and 94 women aged 60 to 85 years, without nutritional deficiencies or illness that could interfere with hematopoiesis, to characterize the normal limits of red and white blood cell counts. Lower limit of hemoglobin normal values were 132 g/l for men and 125 g/l for women. The corresponding figures for MCV were 85 fl and 83 fl for men and women respectively. Lower and upper normal limits for leukocyte count in both sexes were 3.66 and 9.36 x 10(9)/l. Hypersegmented neutrophils existed in 7% of men and 14% of women in spite that folate and vitamin B12 deficiencies were excluded. We concluded that blood cell counts of elderly people have mild deviations of normal values for young adults. Aged humans have a low white cell and bands counts, and elderly women have a higher hemoglobin concentration.

Increased in vitro tumour necrosis factor-alpha production in iron deficiency anemia.

In vitro monocyte-derived tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) production was assessed in iron deficient with anemia (IDA), iron deficient without anemia (ID) and control infants. The concentrations of released and cell-associated cytokines were measured before and after 3 months of iron supplementation in all groups (ferrous sulphate drops: 3 mg/Kg/day). No difference in released and cell-associated IL-1 beta was observed between either groups of infants. Lipopolysaccharide-stimulated blood mononuclear cells from IDA (n = 9) infants produced a significantly higher immunoreactive TNF-alpha concentration as compared to ID (n = 9) and normal subjects (n = 18) on admission (F = 6.72; p < 0.004). After iron therapy, the LPS stimulated TNF-alpha secretion by cells of IDA infants returned to the levels observed in the other groups. Since TNF-alpha plays a key role in iron metabolism, we speculate that increased TNF production in IDA infants could exacerbate the inhibition of erythroid proliferation present in these conditions. Further studies are needed to evaluate the effect of more severe anemia as well as to clarify the biological effect of increased TNF-alpha production in iron deficiency anemia and its consequences.

Effectiveness of iron-fortified infant cereal in prevention of iron deficiency anemia.

BACKGROUND: Iron deficiency continues to be a common problem among infants throughout the world. Iron-fortified formula is effective in preventing iron deficiency but the benefit of iron-fortified cereal is controversial. METHODS: We compared iron-fortified rice cereal to unfortified rice cereal in infants who were exclusively breast-fed for more than 4 months and to iron-fortified formula in infants who were weaned to formula before 4 months of age. The design was double blind in respect to the presence or absence of fortification iron in the cereal or formula and included 515 infants who were followed on the protocol from 4 to 15 months of age. Rice cereal was fortified with 55 mg of electrolytic iron per 100 g of dry cereal and infant formula with 12 mg of ferrous sulfate per 100 g of dry powder, levels approximating those in use in the United States. Measures of iron status were obtained at 8, 12, and 15 months. Infants with hemoglobin levels of < 105 g/L were excluded from the study and treated. RESULTS: Consumption of cereal reached plateaus at means of about 30 g/d after 6 months of age in the formula-fed groups and 26 g/d after 8 months in the breast-fed groups; these amounts are higher than the 19-g/d mean intake by the 73% of infants who consume such cereal in the United States. Among infants weaned to formula before 4 months, the cumulative percentages of infants excluded for anemia by 15 months were 8%, 24%, and 4%, respectively, in the fortified cereal, unfortified cereal and formula, and fortified formula groups (P < .01 unfortified vs either fortified group; the difference between the two fortified groups was not significant). In infants breast-fed for more than 4 months, the corresponding values were 13% and 27%, respectively, in the fortified and unfortified cereal groups (P < .05). Mean hemoglobin level and other iron status measures were in accord with these findings. CONCLUSION: Iron-fortified infant rice cereal can contribute substantially to preventing iron deficiency anemia.

Effect of bovine-hemoglobin-fortified cookies on iron status of schoolchildren: a nationwide program in Chile.

The Chilean School Lunch program, which serves one million children nationwide, was supplied with three 10-g cookies fortified with 6% bovine hemoglobin concentrate, designed to provide 1 mg bioavailable iron per day. A survey of 1000 children was performed after 3 y. Significant differences in hemoglobin concentrations were found in the children from the fortified vs the nonfortified province (P < 0.01). Low serum ferritin values were also significantly more prevalent in the nonfortified group. The effect was evident despite the very low prevalence of anemia in both the fortified and the unfortified school groups. Heme-iron-fortified cookies are a feasible and effective way to improve the iron status of school-age children. In regions of high prevalence of iron-deficiency anemia, the effect of a heme-fortified cookie program should be even more important.