Plant- and Bacteria-Derived Compounds with Anti-Philasterides dicentrarchi Activity.

Philasterides dicentrarchi is a scuticociliate that causes high mortalities in farmed fish. Although vaccination is an effective method to prevent scuticociliatosis caused by the homologous serotype, a universal vaccine has not been developed yet. Many compounds have been shown to be toxic to this ciliate species; moreover, most of them are toxic to aquatic life and cannot be used to prevent the disease. We have evaluated the toxicity to P. dicentrarchi of several compounds of natural origin to be used to reduce parasite levels in the seawater. Ciliates were exposed to several compound concentrations, and the mortality was determined at several incubation times. Tomatine, plumbagin and 2',4'-dihydroxychalcone displayed the highest anticiliate activity, with a dose-dependent response. The effects of these compounds on the EPC cell line were also evaluated, finding that 2',4'-dihydroxychalcone displayed the lowest toxicity to fish cells. At 7.54 µM, 2',4'-dihydroxychalcone inhibited 50% parasite growth but only killed about 10% of EPC cells after 24 h incubation. Finally, we evaluated the toxicity of Pseudomonas H6 surfactant (PS) to P. dicentrarchi, finding that PS was toxic to the ciliate but showed lower toxicity to EPC cells. At a concentration of 7.8 µg/mL (LC50 for the ciliate after 3 h incubation), PS killed 14.9% of EPC cells. We conclude that 2',4'-dihydroxychalcone, and PS could be used to reduce parasite levels in seawater, thus decreasing the risk of scuticociliatosis infection in cultured fish.

Characterization of the turbot Scophthalmus maximus (L.) myeloperoxidase. An insight into the evolution of vertebrate peroxidases.

We have completed the characterization of the turbot (Scophthalmus maximus) myeloperoxidase (mpx) gene and protein, which we partially described in a previous study. The turbot mpx gene has 15 exons that encode a protein of 767 aa, with a signal peptide, propeptide and light and heavy chains, and also with haem cavities, a Ca+2-binding motif and several N- and O-glycosylation sites. The mature protein forms homodimers of about 150 kDa and is very abundant in turbot neutrophils. In addition to the mpx (epx2a) gene, another three peroxidase genes, named epx1, epx2b1 and epx2b2, were identified in the turbot genome. Epx1, Epx2b1 and Epx2b2 proteins also have signal peptides and many structural characteristics of mammalian MPO and eosinophil peroxidase (EPX). Mpx was strongly expressed in head kidney, while epx2b1 and epx2b2 were strongly expressed in the gills, and epx1 was not expressed in any of the tissues or organs analysed. In vitro stimulation of head kidney leucocytes with the parasite Philasterides dicentrarchi caused a decrease in mpx expression and an increase in epx2b1 expression over time. In turbot infected experimentally with P. dicentrarchi a significant increase in mpx expression in the head kidney was observed on day 7 postinfection, while the other genes were not regulated. However, mpx, epx2b1 and epx2b2 were downregulated in the gills of infected fish, and epx1 expression was not affected. These results suggest that the four genes responded differently to the same stimuli. Interestingly, BLAST analysis revealed that Epx1 and Mpx showed greater similarity to mammalian EPX than to MPO. Considering the phylogenetic and synteny data obtained, we concluded that the epx/mpx genes of Gnathostomes can be divided into three main clades: EPX1, which contains turbot epx1, EPX2, which contains turbot mpx (epx2a) and epx2b1 and epx2b2 genes, and a clade containing mammalian EPX and MPO (EPX/MPO). EPX/MPO and EPX2 clades share a common ancestor with the chondrichthyan elephant shark (Callorhinchus milii) and the coelacanth (Latimeria chalumnae) peroxidases. EPX2 was only found in fish and includes two sister groups. One of the groups includes turbot mpx and was only found in teleosts. Finally, the other group contains epx2b1 and epx2b2 genes, and epx2b1-2b2 loci share orthologous genes with other teleosts and also with holosteans, suggesting that these genes appeared earlier on than the mpx gene.

Clinical Outcomes After 1 Year of Augmented Insulin Pump Therapy in Patients with Diabetes in a Specialized Diabetes Center in Medellín, Colombia.

BACKGROUND: Type 1 diabetes can be difficult to control. Augmented pump therapy (CSII-rtCGM) has become an important tool for controlling blood glucose and decreasing hypoglycemia. METHODS: Describe the results 1 year after starting CSII-rtCGM in patients with diabetes in Medellín, Colombia. This is an observational, retrospective study. Patients with type 1 and type 2 diabetes started on CSII-rtCGM between January 2008 and June 2015 were included. Qualitative variables were analyzed as absolute or relative frequencies. Quantitative variables were obtained through central tendency and dispersion according to the normal distribution of the analyzed variable using Kolmogorov-Smirnov. SPSS 19 from IBM was used. RESULTS: Two hundred forty-seven patients were identified, of those 183 were included. The starting HbA1C was 8.7% ± 1.7% and 7.4% ± 0.8% (P < 0.05) 1 year later. 16.5% of patients had been admitted to the hospital before starting CSII-rtCGM, after 1 year the admission rate was 6.0% (P < 0.05). The incidence of severe hypoglycemia at the beginning was 32%, 1 year later it was 7.1%. CONCLUSION: CSII-rtCGM therapy improves glucose control and decreases severe hypoglycemic events and hospital admission rate.

Vaccine-induced modulation of gene expression in turbot peritoneal cells. A microarray approach.

We used a microarray approach to examine changes in gene expression in turbot peritoneal cells after injection of the fish with vaccines containing the ciliate parasite Philasterides dicentrarchi as antigen and one of the following adjuvants: chitosan-PVMMA microspheres, Freund́s complete adjuvant, aluminium hydroxide gel or Matrix-Q (Isconova, Sweden). We identified 374 genes that were differentially expressed in all groups of fish. Forty-two genes related to tight junctions and focal adhesions and/or actin cytoskeleton were differentially expressed in free peritoneal cells. The profound changes in gene expression related to cell adherence and cytoskeleton may be associated with cell migration and also with the formation of cell-vaccine masses and their attachment to the peritoneal wall. Thirty-five genes related to apoptosis were differentially expressed. Although most of the proteins coded by these genes have a proapoptotic effect, others are antiapoptotic, indicating that both types of signals occur in peritoneal leukocytes of vaccinated fish. Interestingly, many of the genes related to lymphocytes and lymphocyte activity were downregulated in the groups injected with vaccine. We also observed decreased expression of genes related to antigen presentation, suggesting that macrophages (which were abundant in the peritoneal cavity after vaccination) did not express these during the early inflammatory response in the peritoneal cavity. Finally, several genes that participate in the inflammatory response were differentially expressed, and most participated in resolution of inflammation, indicating that an M2 macrophage response is generated in the peritoneal cavity of fish one day post vaccination.

Colombian reference growth curves for height, weight, body mass index and head circumference.

AIM: Published Growth studies from Latin America are limited to growth references from Argentina and Venezuela. The aim of this study was to construct reference growth curves for height, weight, body mass index (BMI) and head circumference of Colombian children in a format that is useful for following the growth of the individual child and as a tool for public health. METHODS: Prospective measurements from 27 209 Colombian children from middle and upper socio-economic level families were processed using the generalised additive models for location, scale and shape (GAMLSS). RESULTS: Descriptive statistics for length and height, weight, BMI and head circumference for age are given as raw and smoothed values. Final height was 172.3 cm for boys and 159.4 cm for girls. Weight at 18 years of age was 64.0 kg for boys and 54 kg for girls. Growth curves are presented in a ± 3 SD format using logarithmic axes. CONCLUSION: The constructed reference growth curves are a start for following secular trends in Colombia and are also in the presented layout an optimal clinical tool for health care.

Epidemiological characterization of diabetic patients on therapy with continuous subcutaneous insulin infusion and continuous glucose monitoring in real time.

OBJECTIVE: To describe baseline characteristics of diabetic patients who were started on insulin pump and real time continuous glucose monitor (CSII-rtCGM) in a specialized center in Medellin, Colombia. MATERIALS AND METHODS: All patients with diabetes with complete data who were started on CSII-rtCGM between February 2010 and May 2014 were included. This is a descriptive analysis of the sociodemographic and clinical characteristics. RESULTS: 141 of 174 patients attending the clinic were included. 90,1% had type 1diabetes (T1D). The average age of T1D patients at the beginning of therapy was 31,4 years (SD 14,1). 75.8% of patients had normal weight (BMI<25), 21.0% were overweight (BMI 25-30) and 2,3% were obese (BMI>30). The median duration of T1D was 13 years (P25-P75=10.7-22.0). 14,2% of the patients were admitted at least once in the year preceding the start of CSII-rtCGM because of diabetes related complications. Mean A1c was 8.6%±1.46%. The main reasons for starting CSII-rtCGM were: poor glycemic control (50.2%); frequent hypoglycemia, nocturnal hypoglycemia, hypoglycemia related to exercise, asymptomatic hypoglycemia (30.2%); severe hypoglycemia (16.44%) and dawn phenomena (3.1%). CONCLUSION: Baseline characteristics of patients included in this study who were started on CSII-rtCGM are similar to those reported in the literature. The Clinic starts CSII-rtCGM mainly in T1D patients with poor glycemic control, frequent or severe hypoglycemia despite being on basal/bolus therapy.

Caracterización epidemiológica de pacientes diabéticos en terapia con infusión subcutánea continua de insulina y monitorización continua de glucosa en tiempo real / Epidemiological characterization of diabetic patients on therapy with continuous subcutaneous insulin infusion and continuous glucose monitoring in real time

OBJETIVO: Caracterizar los pacientes con diabetes que iniciaron terapia con bomba de insulina y monitorización continua de glucosa en tiempo real (CSII-rtCGM) en un centro especializado de Medellín, Colombia. MATERIALES Y MÉTODOS: Se evaluaron los pacientes con diabetes que recibieron entrenamiento e instalación del dispositivo en el centro entre febrero de 2010 y mayo de 2014. Se realizó un análisis descriptivo de las variables sociodemográficas y clínicas. RESULTADOS: Se incluyeron 141 pacientes de los 174 pertenecientes al programa. El 90,1% tenía diabetes tipo 1 (DT1), siendo la edad promedio al inicio de la terapia de 31,4 años (SD: 14,1). El 75,8% de los pacientes tenía peso normal (IMC < 25), el 21,0% sobrepeso (IMC = 25-30) y el 2,3% era obeso (IMC ≥ 30). La mediana de duración de la enfermedad fue de 13 años (P25-P75 = 10,7-22,0). El 14,2% de los pacientes fueron hospitalizados al menos una vez en el año previo al inicio de la terapia con CSII-rtCGM por causas relacionadas con la diabetes. El promedio de hemoglobina glucosilada era de 8,6% ± 1,46%. Las principales indicaciones para la formulación de CSII-rtCGM fueron: mal control glucémico (50,2%); hipoglucemias frecuentes, nocturnas o asociadas al ejercicio y sin síntomas de alarma (30,2%); hipoglucemias severas (16,44%) y fenómeno del alba (3,1%). CONCLUSIÓN: Las características de los pacientes que inician terapia con CSII-rtCGM son similares a las reportadas en la literatura. Los resultados muestran que el centro especializado inicia esta terapia principalmente en pacientes con DT1 con mal control glucémico a pesar de la terapia intensiva, hipoglucemias leves persistentes o hipoglucemias severas

OBJECTIVE: To describe baseline characteristics of diabetic patients who were started on insulin pump and real time continuous glucose monitor (CSII-rtCGM) in a specialized center in Medellin, Colombia. MATERIALS AND METHODS: All patients with diabetes with complete data who were started on CSII-rtCGM between February 2010 and May 2014 were included. This is a descriptive analysis of the sociodemographic and clinical characteristics. RESULTS: 141 of 174 patients attending the clinic were included. 90,1% had type 1 diabetes (T1D). The average age of T1D patients at the beginning of therapy was 31,4 years (SD 14,1). 75.8% of patients had normal weight (BMI < 25), 21.0% were overweight (BMI 25-30) and 2,3% were obese (BMI > 30). The median duration of T1D was 13 years (P25-P75 = 10.7-22.0). 14,2% of the patients were admitted at least once in the year preceding the start of CSII-rtCGM because of diabetes related complications. Mean A1c was 8.6% ± 1.46%. The main reasons for starting CSII-rtCGM were: poor glycemic control (50.2%); frequent hypoglycemia, nocturnal hypoglycemia, hypoglycemia related to exercise, asymptomatic hypoglycemia (30.2%); severe hypoglycemia (16.44%) and dawn phenomena (3.1%). CONCLUSION: Baseline characteristics of patients included in this study who were started on CSII-rtCGM are similar to those reported in the literature. The Clinic starts CSII-rtCGM mainly in T1D patients with poor glycemic control, frequent or severe hypoglycemia despite being on basal/bolus therapy

Ecological determinants of the occurrence and dynamics of Vibrio parahaemolyticus in offshore areas.

The life cycle of Vibrio parahaemolyticus has been conventionally associated with estuarine areas characterized by moderate salinity and warm seawater temperatures. Recent evidence suggests that the distribution and population dynamics of V. parahaemolyticus may be shaped by the existence of an oceanic transport of communities of this organism mediated by zooplankton. To evaluate this possibility, the presence of V. parahaemolyticus in the water column of offshore areas of Galicia was investigated by PCR monthly over an 18-month period. Analysis of zooplankton and seawater showed that the occurrence of V. parahaemolyticus in offshore areas was almost exclusively associated with zooplankton and was present in 80% of the samples. The influence of environmental factors assessed by generalized additive models revealed that the abundance and seasonality of V. parahaemolyticus in zooplankton was favoured by the concurrence of downwelling periods that promoted the zooplankton patchiness. These results confirm that offshore waters may be common habitats for V. parahaemolyticus, including strains with virulent traits. Additionally, genetically related populations were found in offshore zooplankton and in estuaries dispersed along 1500 km. This finding suggests that zooplankton may operate as a vehicle for oceanic dispersal of V. parahaemolyticus populations, connecting distant regions and habitats, and thereby producing impacts on the local community demography and the spread of Vibrio-related diseases.

Prevalence and genetic diversity of pathogenic populations of Vibrio parahaemolyticus in coastal waters of Galicia, Spain.

The natural reservoirs and biological characteristics of pathogenic populations of Vibrio parahaemolyticus in marine habitats remain unclear due to difficulties in obtaining pathogenic strains from the environment. The distribution and characteristics of pathogenic V. parahaemolyticus were investigated over 1 year in three coastal environments in Galicia (Spain), including areas of the major international ports in the region. Vibrio parahaemolyticus was present in 35.3% of the samples analysed, and 535 strains were isolated over the period of study. Virulence genes were detected in 94 strains with diverse genetic traits: 66 trh+/tdh-, 24 trh-/tdh+ and 4 trh+/tdh+. Different spatial and seasonal patterns were observed in relation to genetic traits. The trh+/tdh- strains were detected exclusively in northern areas and prevailed in the autumn, when seawater is warmer and less saline, whereas the trh-/tdh+ strains were found in all three areas throughout winter and spring. Characterization of potentially pathogenic strains from the environment revealed an unexpectedly diverse array of serotypes and pulsed-field gel electrophoresis (PFGE) profiles (pulsotypes) that were unrelated to clinical strains of V. parahaemolyticus that are prevalent in Spain. The results of the current study provide a novel view of V. parahaemolyticus in Europe, in which diverse pathogenic groups are constitutive components of the environmental populations in coastal habitats.

Anti-infectious antibodies and autoimmune-associated autoantibodies in patients with type I diabetes mellitus and their close family members.

Type 1 diabetes mellitus (T1DM) is an autoimmune disease with complex interactions between genetic and environmental factors. We compared antibody levels to various infectious agents and of autoimmune-associated autoantibodies between Colombian T1DM patients, their close family members and healthy controls. Significantly lower levels of antibodies against several infectious agents were detected in the T1DM patients. These included Helicobacter pylori (P = 0.01), cytomegalovirus (P = 0.001), Epstein-Barr virus (P = 0.02) and Toxoplasma (P = 0.001). T1DM patients had significantly higher levels of IgG-anti-gliadin antibodies (P = 0.001) and IgG-antitissue transglutaminase antibodies (P = 0.03), and a borderline association with anticentromere antibodies (P = 0.06). The lower level of antibodies against infectious agents in T1DM patients may be related to their younger ages, but may also point to a protective role of those infections in T1DM development in susceptible individuals. Our results confirm the association between T1DM and celiac disease. A possible association with anticentromere antibody needs further studies.

Mucopolisacaridosis tipo IV como causa de talla baja patológica: reporte de un caso y revisión de la literatura / Mucopolysaccharidosis type IV as a cause of pathological short stature: a case report and literature review

Se presenta un caso clínico de síndrome de Morquio o mucopolisacaridosis tipo IV, en una niña de 5 años que consulta al servicio de endocrinología pediátrica del Hospital Pablo Tabón Uribe, por talla baja y deformidades esqueléticas que iniciaron al año de edad. Inicialmente admitida con una impresión diagnóstica de raquitismo, pero al evaluar el caso en conjunto con el grupo de ortopedia infantil se confirma el diagnóstico por clínica, hallazgos radiológicos característicos y pruebas específicas para mucopolisacaridosis. Se revisan los principales aspectos clínicos y radiológicos de la enfermedad y el manejo actual.

We report a case of Morquio syndrome, or mucopolysaccharidosis type IV, in a girl of 5 years attending the pediatric endocrinology service at the Pablo Tobón Uribe Hospital, because of short stature and skeletal deformities that began in the first year of life. Initially admitted with a working diagnosis of raquitism, but reassessment of the case by the children's orthopedic group confirmed the diagnosis by clinical, specific serological tests and characteristical radiological findings specific for mucopolysaccharidosis. We review the clinical and radiological characteristics of the disease and current treatment options.

Emergence of Asiatic Vibrio diseases in South America in phase with El Niño.

BACKGROUND: The seventh pandemic of Vibrio cholerae unexpectedly reached the coast of Peru in 1991, causing an explosive emergence of infections throughout the American continents. The origin and routes of dissemination are as yet unknown. A new Vibrio epidemic arose in 1997 in South America (northern Chile) when the pandemic clone of Vibrio parahaemolyticus was for the fist time detected outside of Asia. These 2 cases were concurrent with 2 episodes of El Niño. METHODS: We carried out a survey of records of V. parahaemolyticus infection and of strains existing in the Instituto Nacional de Salud of Peru between 1994 and 2005. Association between the El Niño event and the V. parahaemolyticus disease was analyzed through generalized additive models applied to time-series data with negative binomial response, selecting some oceanographic factors distinctive of the movement of the El Niño waters. RESULTS: Epidemiologic data and laboratory investigations of the strains showed that V. parahaemolyticus infections caused by the pandemic clone emerged in the coasts of Peru linked to the 1997 El Niño episode. The epidemic dissemination of this clone matched the expansion and dynamics of the poleward propagation and the receding of the El Niño waters. This pattern was similar to previously reported onset of cholera epidemic in 1991. CONCLUSIONS: These findings identify the El Niño episodes as a reliable vehicle for the introduction and propagation of Vibrio pathogens in South America. The movement of oceanic waters seems to be one of the driving forces of the spread of Vibrio diseases.

Effects of child- and adolescent-onset endogenous Cushing syndrome on bone mass, body composition, and growth: a 7-year prospective study into young adulthood.

UNLABELLED: The long-term effects on bone and fat mass in children with endogenous CS are unknown. In 14 children followed for 3-7 years into young adulthood after cure of CS, whereas bone mass largely recovered, persisting increases in total body and visceral fat suggests an increase risk of the metabolic syndrome. INTRODUCTION: Endogenous Cushing syndrome (CS) is associated with decreased bone mass and increased central fat mass. Whereas bone mass seems to improve after successful treatment, little is known about whether central fat persists. MATERIALS AND METHODS: This was a prospective study of 14 children (10 girls and 4 boys) and adolescents with CS who were successfully treated and remained eucortisolemic. Growth, puberty, bone mass, and body composition were evaluated at baseline and during regular follow-up for 3 years and in seven children for a further 4 years of remission to assess final adult height (FH), BMI, bone mass, and body composition. RESULTS: CS compromised growth, leading to about a -0.8 SD loss of FH and 0.9 SD increase in weight and BMI. BMD apparent density (BMAD) SD Score (SDS) at the lumbar spine (LS) at diagnosis were -1.8 and -1.25, respectively, and after 3 years of follow-up approached the mean with no further increase apparent up to 7 years of follow-up. Whereas hip BMD SDS increased from -1.3 at diagnosis to -0.40 at 3 years and 0 at 7 years of follow-up, femoral neck BMAD remained at or around 0 SDS at diagnosis and during follow-up. BMI was >25 kg/m(2) in five of seven adult subjects, most of whom were women. Total body fat and the ratio of visceral to subcutaneous was abnormally high in the majority of these subjects, whereas LS volumetric BMD was -0.7 SDS. CONCLUSIONS: Despite remission of CS, children and adolescents have significant alterations in body composition that result in a small but significant decrease in bone mass and increase in visceral adiposity. Although bone mass largely recovers after endogenous CS, changes in total and visceral fat suggest these subjects are at increased risk of the metabolic syndrome. Therefore, long-term monitoring of body fat and bone mass is mandatory after treatment of CS.

Familial clustering of autoimmune diseases in patients with type 1 diabetes mellitus.

We investigated the familial aggregation of autoimmune diseases (AIDs) among first-degree relatives (FDR) of patients with type 1 diabetes mellitus (T1D). Relatives of 98 T1D patients defined according to the guidelines diagnosis of the American Diabetes Association and 113 matched controls without any AID, were interviewed using a questionnaire that sought information about demographic and medical characteristics including a list of 18 AIDs. Genetic analysis was performed using the program ASSOC and by calculating recurrent risk ratios. In cases, 25.5% of the families had at least one member having an AID, while in controls there were 9% (odds ratio [OR]: 3.96, 95% confidence interval [CI]=1.74-9.0, p=0.0006). An AID was registered in 8.3% of 312 FDR of patients as compared with 2.4% of 362 FDR in controls (OR: 3.56, 95% CI=1.64-7.73, p=0.0008). The most frequent AIDs registered in FDR of cases were autoimmune thyroid disease (AITD) and T1D, which disclosed coefficients of aggregation. These results indicate that AIDs cluster within families of T1D patients adding further evidence to consider that clinically different autoimmune phenotypes may share common susceptibility gene variants, which may act pleiotropically as risk factors for autoimmunity.

The role of the resting zone in growth plate chondrogenesis.

In mammals, growth of long bones occurs at the growth plate, a cartilage structure that contains three principal layers: the resting, proliferative, and hypertrophic zones. The function of the resting zone is not well understood. We removed the proliferative and hypertrophic zones from the rabbit distal ulnar growth plate in vivo, leaving only the resting zone. Within 1 wk, a complete proliferative and hypertrophic zone often regenerated. Next, we manipulated growth plates in vivo to place resting zone cartilage ectopically alongside the proliferative columns. Ectopic resting zone cartilage induced a 90-degree shift in the orientation of nearby proliferative zone chondrocytes and seemed to inhibit their hypertrophic differentiation. Our findings suggest that resting zone cartilage makes important contributions to endochondral bone formation at the growth plate: 1) it contains stem-like cells that give rise to clones of proliferative chondrocytes; 2) it produces a growth plate-orienting factor, a morphogen, that directs the alignment of the proliferative clones into columns parallel to the long axis of the bone; and 3) it may also produce a morphogen that inhibits terminal differentiation of nearby proliferative zone chondrocytes and thus may be partially responsible for the organization of the growth plate into distinct zones of proliferation and hypertrophy.

Recovery from osteoporosis through skeletal growth: early bone mass acquisition has little effect on adult bone density.

It is often assumed that bone mineral accretion should be optimized throughout childhood to maximize peak bone mass. In contrast, we hypothesized that bone mineral acquisition early in life would have little or no effect on adult bone mass because many areas of the juvenile skeleton are replaced in toto through skeletal growth. To test this hypothesis, we induced osteoporosis by administering dexamethasone to 5-week-old rabbits for 5 weeks and then allowed them to recover for 16 weeks. Tibial bone mineral density (ash weight/volume) was decreased in the dexamethasone-treated animals at the end of treatment but recovered completely. Bone structure in the femur was assessed by histomorphometry. Trabecular and cortical bone in the distal metaphysis was made osteoporotic by dexamethasone, but was then replaced through endochondral bone formation and recovered. Periosteal bone formation rate in the diaphysis was decreased during dexamethasone treatment but afterwards rebounded above controls and normalized cortical width. Our data suggest that bone mineral acquisition early in life has little effect on adult bone density because the juvenile bone is largely replaced through growth. If this concept generalizes, then interventions to maximize peak bone mass should be directed at adolescents rather than young children.