Indirect effects of emotion regulation in the relationship between pain and cannabis use in adults 18-64 years.

INTRODUCTION: Individuals with chronic pain often receive prescription opioid medication, and they may use cannabis to treat pain as well, although the risks of cannabis-opioid co-use are significant. This study aimed to investigate whether two transdiagnostic factors, emotion regulation and distress tolerance, had significant indirect effects in the relationship between pain and cannabis use in adults with chronic pain and an opioid prescription. METHODS: Participants (n = 450; mean age = 38.6 ± 11.09) were recruited using Qualtrics panel service and were 75 % female and 79 % White, non-Hispanic. Participants completed a 30-minute self-report survey capturing three-month cannabis use, the Difficulties in Emotional Regulation Scale (DERS), and the Distress Tolerance Scale (DTS). The Graded Pain Scale (GCPS) assessed pain severity/intensity and disability. Analyses used the SPSS PROCESS macro, with both single (i.e., one transdiagnostic factor) and parallel indirect effects (i.e., both the DERS and DTS) examined. RESULTS: There were statistically significant indirect effects for both the DERS and DTS in the relationship between pain intensity or disability and three-month cannabis use in single factor models. In the parallel indirect effect model, only the DERS was statistically significant (intensity indirect effect coefficient = 0.0195 % confidence interval [95 %CI] = 0.0065, 0.390; disability indirect effect coefficient = 0.0147, 95 %CI = 0.0055, 0.0274). CONCLUSIONS: When examining parallel indirect effects, only emotional regulation and not distress tolerance mediated the relationship between chronic pain and cannabis use among those with an opioid prescription. Clinically, interventions aimed at improving emotional regulation in individuals with chronic pain can help limit cannabis and opioid co-use.

El sicariato: una mirada al fenómeno desde la perspectiva criminológica / Contract assasination: a criminological perspective on the phenomenon / Sicariato: uma perspectiva criminológica sobre o fenômeno

Antecedentes: el sicariato es un fenómeno de violencia que afecta a la sociedad contemporánea, en medio de valores relativos al respeto de la vida y la integridad humana. En Colombia, los datos y cifras de los homicidios evidencian que durante los años 2017-2020 se presentaron aproximadamente 26 161 casos en la modalidad sicariato; las motivaciones se asocian fundamentalmente al narcotráfico, economías ilegales, disputas territoriales y factores sociales. Objetivo: identificar la etimología de la palabra "sicario", características, investigaciones y algunas tendencias generacionales sobre el sicariato. Método: este estudio se realizó desde un enfoque cualitativo, diseño documental mediante la revisión de algunas teorías sobre subculturas criminales (Cohen, 1955), el aprendizaje de la conducta delictiva (Sutherland, 1974), aprendizaje social (Bandura, 1987; Becker, 1974), crimen y castigo (Bourdieu, 1997), el entorno social que lo rodea y economía criminal, otros autores y revisión bases de datos: Mendeley, Proquest, Redalyc, base, BBC learning english, entre otros. Resultados: este trabajo permitió recoger conceptos, características, tendencias delictivas y análisis criminológicos del fenómeno; se establecen algunas tipologías, por modus operandi y capacidad del perpetrador del hecho punible, surgimiento del sicariato en Colombia y algunas características de los códigos de conducta de las dos últimas generaciones de sicarios. Finalmente, se plantean las conclusiones.

Background: hired assassination is a phenomenon of violence that affects contemporary society, amidst values related to respect for life and human integrity. In Colombia, homicide data and figures show that during the years 2017-2020 there were approximately 26 161 cases in the form of hired killings; the motivations mainly associated with drug trafficking, illegal economies, territorial disputes and social factors. Objective: to identify the etymology of the word "sicario", characteristics, research and some generational trends regarding contract killing. Method: this study was conducted based on a qualitative approach, documentary design by reviewing several theories on criminal subcultures (Cohen, 1955), learning about criminal behaviour (Sutherland, 1974), social learning (Bandura, 1987; Becker, 1974), crime and punishment (Bourdieu, 1997) and the surrounding social environment and criminal economy, studying other authors and reviewing the databases: Mendeley, Proquest, Redalyc, base, BBC learning English, among others. Results: this work allowed the collection of concepts, characteristics, criminal trends and criminological analysis of the phenomenon; some typologies are established, by modus operandi and capacity of the perpetrator of the punishable act, the emergence of assasination by contract in Colombia and some characteristics of the codes of conduct of the last two generations of hitmen. Finally, conclusions are drawn.

Antecedentes: o sicariato ou o assassinato contratado é um fenômeno de violência que afeta a sociedade contemporânea, em meio a valores relacionados ao respeito à vida e à integridade humana. Na Colômbia, os dados e números de homicídios mostram que, durante 2017 e 2020, houve aproximadamente 26 161 casos na forma de sicariato ou assassinatocontratado; as motivações estão associadas principalmente ao tráfico de drogas, economias ilegais, disputas territoriais e fatores sociais. Objetivo: identificar a etimologia da palavra "sicário", características, pesquisas e algumas tendências geracionais sobre o sicariato. Método: este estudo foi realizado a partir de uma abordagem qualitativa, com desenho documental, por meio da revisão de algumas teorias sobre subculturas criminais (Cohen, 1955), aprendizagem do comportamento criminoso (Sutherland, 1974), aprendizagem social (Bandura, 1987; Becker, 1974), crime e castigo (Bourdieu, 1997), ambiente social circundante e economia criminal, outros autores e bancos de dados de revisão: Mendeley, Proquest, Redalyc, base, BBC learning english, entre outros. Resultados: este trabalho permitiu a coleta de conceitos, características, tendências criminais e análise criminológica do fenômeno; são estabelecidas algumas tipologias, por modus operandi e capacidade do autor do ato punível, o surgimento do sicariato na Colômbia e algumas características dos códigos de conduta das duas últimas gerações de sicários. Por fim, são apresentadas as conclusões.

In Vitro Antifungal Activity of LL-37 Analogue Peptides against Candida spp.

Fungal infections have increased in recent decades with considerable morbidity and mortality, mainly in immunosuppressed or admitted-to-the-ICU patients. The fungal resistance to conventional antifungal treatments has become a public health problem, especially with Candida that presents resistance to several antifungals. Therefore, generating new alternatives of antifungal therapy is fundamental. One of these possibilities is the use of antimicrobial peptides, such as LL-37, which acts on the disruption of the microorganism membrane and promotes immunomodulatory effects in the host. In this study, we evaluated the in vitro antifungal activity of the LL-37 analogue peptides (AC-1, LL37-1, AC-2, and D) against different Candida spp. and clinical isolates obtained from patients with vulvovaginal candidiasis. Our results suggest that the peptides with the best ranges of MICs were LL37-1 and AC-2 (0.07 µM) against the strains studied. This inhibitory effect was confirmed by analyzing the yeast growth curves that evidenced a significant decrease in the fungal growth after exposure to LL-37 peptides. By the XTT technique we observed a significant reduction in the biofilm formation process when compared to yeasts untreated with the analogue peptides. In conclusion, we suggest that LL-37 analogue peptides may play an important antimicrobial role against Candida spp.

Silent but Not Harmless: A Synonymous SLC5A5 Gene Variant Leading to Dyshormonogenic Congenital Hypothyroidism.

Background: Congenital iodide transport defect (ITD) is an uncommon cause of dyshormonogenic congenital hypothyroidism characterized by the absence of active iodide accumulation in the thyroid gland. ITD is an autosomal recessive disorder caused by loss-of-function variants in the sodium/iodide symporter (NIS)-coding SLC5A5 gene. Objective: We aimed to identify, and if so to functionally characterize, novel ITD-causing SLC5A5 gene variants in a cohort of five unrelated pediatric patients diagnosed with dyshormonogenic congenital hypothyroidism with minimal to absent 99mTc-pertechnetate accumulation in the thyroid gland. Methods: The coding region of the SLC5A5 gene was sequenced using Sanger sequencing. In silico analysis and functional in vitro characterization of a novel synonymous variant were performed. Results: Sanger sequencing revealed a novel homozygous synonymous SLC5A5 gene variant (c.1326A>C in exon 11). In silico analysis revealed that the c.1326A>C variant is potentially deleterious for NIS pre-mRNA splicing. The c.1326A>C variant was predicted to lie within a putative exonic splicing enhancer reducing the binding of splicing regulatory trans-acting protein SRSF5. Splicing minigene reporter assay revealed that c.1326A>C causes exon 11 or exon 11 and 12 skipping during NIS pre-mRNA splicing leading to the NIS pathogenic variants p.G415_P443del and p.G415Lfs*32, respectively. Significantly, the frameshift variant p.G415Lfs*32 is predicted to be subjected to degradation by nonsense-mediated decay. Conclusions: We identified the first exonic synonymous SLC5A5 gene variant causing aberrant NIS pre-mRNA splicing, thus expanding the mutational landscape of the SLC5A5 gene leading to dyshormonogenic congenital hypothyroidism.

A Novel SLC5A5 Variant Reveals the Crucial Role of Kinesin Light Chain 2 in Thyroid Hormonogenesis.

CONTEXT: Iodide transport defect (ITD) (Online Mendelian Inheritance in Man No. 274400) is an uncommon cause of dyshormonogenic congenital hypothyroidism due to loss-of-function variants in the SLC5A5 gene, which encodes the sodium/iodide symporter (NIS), causing deficient iodide accumulation in thyroid follicular cells. OBJECTIVE: This work aims to determine the molecular basis of a patient's ITD clinical phenotype. METHODS: The propositus was diagnosed with dyshormonogenic congenital hypothyroidism with minimal 99mTc-pertechnetate accumulation in a eutopic thyroid gland. The propositus SLC5A5 gene was sequenced. Functional in vitro characterization of the novel NIS variant was performed. RESULTS: Sanger sequencing revealed a novel homozygous missense p.G561E NIS variant. Mechanistically, the G561E substitution reduces iodide uptake, because targeting of G561E NIS to the plasma membrane is reduced. Biochemical analyses revealed that G561E impairs the recognition of an adjacent tryptophan-acidic motif by the kinesin-1 subunit kinesin light chain 2 (KLC2), interfering with NIS maturation beyond the endoplasmic reticulum, and reducing iodide accumulation. Structural bioinformatic analysis suggests that G561E shifts the equilibrium of the unstructured tryptophan-acidic motif toward a more structured conformation unrecognizable to KLC2. Consistently, knockdown of Klc2 causes defective NIS maturation and consequently decreases iodide accumulation in rat thyroid cells. Morpholino knockdown of klc2 reduces thyroid hormone synthesis in zebrafish larvae leading to a hypothyroid state as revealed by expression profiling of key genes related to the hypothalamic-pituitary-thyroid axis. CONCLUSION: We report a novel NIS pathogenic variant associated with dyshormonogenic congenital hypothyroidism. Detailed molecular characterization of G561E NIS uncovered the significance of KLC2 in thyroid physiology.

Toxicity of three organophosphate insecticides in anuran embryos at different temperatures / Toxicidad de tres insecticidas organofosforados en embriones de anuros a diferentes temperaturas

RESUMEN Los efectos de diferentes agroquímicos y de la temperatura han sido temas recurrentes en la investigación en anuros; sin embargo, estas variables se han abordado de manera independiente sin considerar que pueden ejercer una presión simultánea sobre las especies. Por esta razón, este trabajo tiene como objetivo determinar la toxicidad (a través de la Concentración Letal Media -CL50) de los insecticidas organofosforados clorpirifos, diazinón y monocrotofos, bajo tres regímenes térmicos (23, 28 y 33 ± 1 °C) sobre embriones de tres especies de anuros. De acuerdo a los valores de CL50, el insecticida clorpirifos fue el más tóxico, seguido del diazinón y del monocrotofos. Por su parte, de manera general se encontró un incremento de la toxicidad de los insecticidas organofosforados a la temperatura más alta de experimentación (33 °C). Además, el efecto de la temperatura se hizo más notorio para los organismos expuestos al clorpirifos, el insecticida más letal. Estos resultados sugieren un efecto negativo para la fauna acuática de anuros debido al actual uso desmesurado de este tipo de agroquímicos y a su interacción con la temperatura ambiental.

ABSTRACT In anurans, the effects of different agrochemicals and temperature have been recurrent topics in research; however, these variables are addressed independently without considering that they can exert a simultaneous pressure on the species. For this reason, this work aims to determine the toxicity (through the Medium Lethal Concentration -LC50) of the organophosphate insecticides: chlorpyrifos, diazinon and monocrotophos, under three thermal regimes (23, 28 and 33 ± 1 °C) on embryos ofthree species of anurans. According to the LC50 values, the chlorpyrifos insecticide was the most toxic, followed by diazinon and monocrotophos. On the other hand, an increase in the toxicity of the organophosphate insecticides studied was generally found at the highest experimental temperature (33 °C). In addition, the effect of temperature became more noticeable for the organisms exposed to chlorpyrifos, the most lethal insecticide. These results suggest a negative effect for the aquatic fauna of anurans due to the current excessive use of this type of agrochemicals and their interaction with environmental temperature.

Publisher Correction: Morphological, genotypic and metabolomic signatures confirm interfamilial hybridization between the ubiquitous kelps Macrocystis (Arthrothamnaceae) and Lessonia (Lessoniaceae).

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Morphological, genotypic and metabolomic signatures confirm interfamilial hybridization between the ubiquitous kelps Macrocystis (Arthrothamnaceae) and Lessonia (Lessoniaceae).

Macrocystis pyrifera and Lessonia spicata are economically and ecologically relevant brown seaweeds that recently have been classified as members of two separated families within Laminariales (kelps). Here we describe for the first time the Macrocystis pyrifera x Lessonia spicata hybridization in the wild (Chiloe Island, Southeastern Pacific), where populations of the two parents exist sympatrically. Externally, this hybrid exhibited typical features of its parents M. pyrifera (cylindrical and flexible distal stipes, serrate frond margins and presence of sporophylls) and L. spicata (rigid and flat main stipe and first bifurcation), as well as intermediate features between them (thick unfused haptera in the holdfast). Histological sections revealed the prevalence of mucilage ducts within stipes and fronds (absent in Lessonia) and fully developed unilocular sporangia in the sporophylls. Molecular analyses confirmed the presence of the two parental genotypes for ITS1 nrDNA and the M. pyrifera genotype for two predominantly maternally inherited cytoplasmic markers (COI and rbcLS spacer) in the tissue of the hybrid. A metabolome-wide approach revealed that this hybrid is more chemically reminiscent to M. pyrifera. Nevertheless, several hits were identified as Lessonia exclusive or more remarkably, not present in any of the parent. Meiospores developed into apparently fertile gametophytes, which gave rise to F1 sporophytes that reached several millimeters before suddenly dying. In-vitro reciprocal crossing of Mar Brava gametophytes from both species revealed that although it is rare, interfamilial hybridization between the two species is possible but mostly overcome by pseudogamy of female gametophytes.

Results of a randomized, double-blind phase II clinical trial of NY-ESO-1 vaccine with ISCOMATRIX adjuvant versus ISCOMATRIX alone in participants with high-risk resected melanoma.

BACKGROUND: To compare the clinical efficacy of New York Esophageal squamous cell carcinoma-1 (NY-ESO-1) vaccine with ISCOMATRIX adjuvant versus ISCOMATRIX alone in a randomized, double-blind phase II study in participants with fully resected melanoma at high risk of recurrence. METHODS: Participants with resected stage IIc, IIIb, IIIc and IV melanoma expressing NY-ESO-1 were randomized to treatment with three doses of NY-ESO-1/ISCOMATRIX or ISCOMATRIX adjuvant administered intramuscularly at 4-week intervals, followed by a further dose at 6 months. Primary endpoint was the proportion free of relapse at 18 months in the intention-to-treat (ITT) population and two per-protocol populations. Secondary endpoints included relapse-free survival (RFS) and overall survival (OS), safety and NY-ESO-1 immunity. RESULTS: The ITT population comprised 110 participants, with 56 randomized to NY-ESO-1/ISCOMATRIX and 54 to ISCOMATRIX alone. No significant toxicities were observed. There were no differences between the study arms in relapses at 18 months or for median time to relapse; 139 vs 176 days (p=0.296), or relapse rate, 27 (48.2%) vs 26 (48.1%) (HR 0.913; 95% CI 0.402 to 2.231), respectively. RFS and OS were similar between the study arms. Vaccine recipients developed strong positive antibody responses to NY-ESO-1 (p≤0.0001) and NY-ESO-1-specific CD4+ and CD8+ responses. Biopsies following relapse did not demonstrate differences in NY-ESO-1 expression between the study populations although an exploratory study demonstrated reduced (NY-ESO-1)+/Human Leukocyte Antigen (HLA) class I+ double-positive cells in biopsies from vaccine recipients performed on relapse in 19 participants. CONCLUSIONS: The vaccine was well tolerated, however, despite inducing antigen-specific immunity, it did not affect survival endpoints. Immune escape through the downregulation of NY-ESO-1 and/or HLA class I molecules on tumor may have contributed to relapse.

Auranofin improves overall survival when combined with standard of care in a pilot study involving dogs with osteosarcoma.

Osteosarcoma is the most common paediatric primary bone malignancy. The major cause of death in osteosarcoma is drug-resistant pulmonary metastasis. Previous studies have shown that thioredoxin reductase 2 is a driver of metastasis in osteosarcoma and can be inhibited by auranofin (AF). Moreover, studies have shown that AF significantly reduces pulmonary metastases in xenotransplant models. Here, we describe a phase I/II study of AF in canine osteosarcoma, a well-recognized spontaneous model of human osteosarcoma. We performed a single-arm multicentre pilot study of AF in combination with standard of care (SOC) (amputation + carboplatin). We recruited 40 dogs to the trial and used a historical SOC-only control group (n = 26). Dogs >15 kg received 9 mg AF q3d PO and dogs <15 kg received 6 mg q3d. Follow-up occurred over at least a 3-year period. Auranofin plus SOC improved overall survival (OS) (P = .036) in all dogs treated. The improved outcome was attributable entirely to improved OS in male dogs (P = .009). At the time of writing, 10 dogs (25%) survive without measurable disease in the treatment group with survival times ranging between 806 and 1525 days. Our study shows that AF improves OS in male dogs when combined with SOC. Our findings have translational relevance for the management of canine and human osteosarcoma. Our data justify a larger multicentre phase 2 trial in dogs and a phase I/II trial in human patients with refractory disease at the time of initial surgery.

Detección de Treponema pallidum subespecie pallidum para el diagnóstico de sífilis congénita mediante reacción en cadena de la polimerasa anidada.

Introducción. La sífilis es una enfermedad producida por Treponema pallidum subespecie pallidum cuya incidencia mundial es de 12 millones de casos por año, aproximadamente; de estos, más de dos millones se presentan en mujeres gestantes, siendo la sífilis congénita la complicación más grave de esta infección en el embarazo.Objetivo. Detectar la presencia de T. pallidum subespecie pallidum en muestras clínicas para el diagnóstico de sífilis congénita mediante reacción en cadena de la polimerasa (PCR) anidada y determinar su concordancia con las pruebas serológicas.Materiales y métodos. Mediante PCR convencional y anidada, se amplificaron tres genes diana (polA, 16S ADNr y TpN47) y se confirmaron los productos de amplificación de los genes TpN47 y polA por secuenciación. Las pruebas serológicas empleadas fueron la VDRL (Venereal Disease Research Laboratory), la de reagina plasmática rápida (Rapid Plasma Reagin, RPR) y la de aglutinación de partículas para Treponema pallidum (Treponema pallidum Particle Agglutination Assay, TPPA).Resultados. La sensibilidad para la PCR convencional fue de 52 pg y, para la PCR anidada, de 0,52 pg. La especificidad con los iniciadores TpN47 y polA fue de 100 %; los resultados de la secuenciación mostraron una identidad de 97 % con T. pallidum. En 70 % de las muestras, los resultados de las pruebas serológicas y la PCR anidada concordaron.Conclusión. El gen TpN47 resultó ser el mejor blanco molecular para la identificación de T. pallidum. La PCR anidada se presenta como una alternativa de diagnóstico molecular promisoria para el diagnóstico de sífilis congénita.

Detección de Treponema pallidum subespecie pallidum para el diagnóstico de sífilis congénita mediante reacción en cadena de la polimerasa anidada / Detection of Treponema pallidum subspecies pallidum for the diagnosis of congenital syphilis by nested polymerase chain reaction

Resumen Introducción. La sífilis es una enfermedad producida por Treponema pallidum subespecie pallidum cuya incidencia mundial es de 12 millones de casos por año, aproximadamente; de estos, más de dos millones se presentan en mujeres gestantes, siendo la sífilis congénita la complicación más grave de esta infección en el embarazo. Objetivo. Detectar la presencia de T. pallidum subespecie pallidum en muestras clínicas para el diagnóstico de sífilis congénita mediante reacción en cadena de la polimerasa (PCR) anidada y determinar su concordancia con las pruebas serológicas. Materiales y métodos. Mediante PCR convencional y anidada, se amplificaron tres genes diana (polA, 16S ADNr y TpN47) y se confirmaron los productos de amplificación de los genes TpN47 y polA por secuenciación. Las pruebas serológicas empleadas fueron la VDRL (Venereal Disease Research Laboratory), la de reagina plasmática rápida (Rapid Plasma Reagin, RPR) y la de aglutinación de partículas para Treponema pallidum (Treponema pallidum Particle Agglutination Assay, TPPA). Resultados. La sensibilidad para la PCR convencional fue de 52 pg y, para la PCR anidada, de 0,52 pg. La especificidad con los iniciadores TpN47 y polA fue de 100 %; los resultados de la secuenciación mostraron una identidad de 97 % con T. pallidum. En 70 % de las muestras, los resultados de las pruebas serológicas y la PCR anidada concordaron. Conclusión. El gen TpN47 resultó ser el mejor blanco molecular para la identificación de T. pallidum. La PCR anidada se presenta como una alternativa de diagnóstico molecular promisoria para el diagnóstico de sífilis congénita.

Abstract Introduction. Syphilis is a disease produced by Treponema pallidum subspecies pallidum, which affects approximately 12 million people worldwide every year. Of these, more than 2 million are pregnant women whose babies end up having congenital syphilis, the worst form of this infection. Objective. To detect the presence of T. pallidum subspecies pallidum in clinical samples in order to diagnose congenital syphilis by means of nested PCR, and to determine its concordance with serological testing. Materials and methods. Three target genes (polA, 16S ADNr y TpN47) were amplified by conventional and nested PCR. The results from the amplification of the TpN47 and polA genes were confirmed by sequencing. The serological tests used were VDRL (Venereal Disease Research Laboratory), RPR (Rapid Plasma Reagin) y TPPA (Treponema pallidum Particle Agglutination Assay). Results. The sensitivity for the conventional PCR was 52 pg and 0.52 pg for the nested PCR. The specificity of primers TpN47 and polA was 100 %; the results of the sequencing showed a 97 % identity with T. pallidum. There was concordance between the serology and the nested PCR in 70% of the samples. Conclusion. The TpN47 gene was the best molecular target for the identification of T. pallidum. The nested PCR is a promising molecular tool for the diagnosis of congenital syphilis.

Síndrome de interrupción del tallo pituitario: Relaciones clínicas, bioquímicas y neurorradiológicas / Pituitary stalk interruption syndrome: Clinical, biochemical and neuroradiological relationships

El síndrome de interrupción del tallo pituitario (PSIS) se caracteriza por la demostración neurorradiológica de un tallo pituitario ausente, interrumpido o hipoplásico, adenohipófisis aplásica/hipoplásica o neurohipófisis ectópica. Este síndrome se ha relacionado con formas severas de hipopituitarismo congénito (HPC), asociado a múltiples deficiencias de hormonas pituitarias (MPHD). Evaluamos a pacientes con HPC y PSIS, analizando los signos y los síntomas neonatales al diagnóstico, relacionándolos con las deficiencias hormonales pituitarias y signos neurorradiológicos presentes. Estudiamos retrospectivamente a 80 pacientes asistidos en el Hospital de Niños de Córdoba, con diagnóstico de HPC, de los cuales 42 (52%) presentaron PSIS; 22 mujeres y 20 varones, EC: 5 días-9,5 años. El 62% presentó MPHD y el 38% insuficiencia somatotrófica aislada (IGHD). El análisis de las variables perinatales demostró antecedentes de parto natural en el 52% (11/21) de las MPHD vs. 13% (2/15) de las IGHD. Cuatro pacientes, 2 con MPHD y 2 con IGHD presentaban antecedentes obstétricos consistentes en presentación podálica y transversa respectivamente, todos ellos resueltos mediante operación cesárea. Los signos y los síntomas perinatales fueron hipo- glucemia: 61% en MPHD vs. 19% en IGHD, p: 0,0105; ictericia: 38% en MPHD vs. 25% en IGHD; micropene: 77% en MPHD y colestasis: 19% en MPHD. Convulsiones neonatales se presentaron en el 75% de los niños con MPHD e hipoglucemia. EC media de consulta: 2,1 años en MPHD (30% en el período neonatal, 70% antes de 2 años) y 3,6 años en IGHD (44% en menores de 2 años). Los pacientes con MPHD presentaban: tallo no visible 81% (n: 21/26) vs. tallo hipoplásico: 19% (n: 5/26), p: 0,0001; en IGHD 56% (n: 9/16) vs. 44% (n: 7/16), p: 0,5067, respectivamente. El 100% de los neonatos con HPC tenían tallo pituitario ausente. Concluimos que la demostración de PSIS en niños con HPC proporciona información valiosa como predictor de la severidad fenotípica, la presencia de MPHD y de la respuesta al tratamiento. La baja frecuencia de antecedentes obstétricos posicionales potencialmente distócicos, como parte de los mecanismos fisiopatogénicos responsables de PSIS, indicaría la necesidad de analizar la importancia de posibles factores genéticos y epigenéticos involucrados. El diagnóstico precoz de HPC debe sospecharse en presencia de signos y síntomas clínicos, tales como hipoglucemia, colestasis, micropene y defectos asociados en la línea media facial. La resonancia magnética cerebral debe formar parte de los estudios complementarios en pacientes con esta presunción diagnóstica, especialmente a edades tempranas. El reconocimiento tardío de esta entidad puede aumentar la morbilidad y la mortalidad con efectos potenciales deletéreos y permanentes.

Pituitary stalk interruption syndrome (PSIS) is characterised by the combination of an interrupted or thin pituitary stalk, absent or ectopic posterior pituitary, and anterior pituitary hypoplasia. It is manifested as isolated (IGHD) or combined pituitary hormone deficiencies (CPHD) of variable degrees and timing of onset, with a wide spectrum of clinical phenotypes. PSIS may be an isolated morphological abnormality or be part of a syndrome. A retrospective evaluation is presented of clinical signs and symptoms present at early life stages, as well as an analysis of their relationship with hormone laboratory tests and diagnostic imaging in children with congenital hypopituitarism (CHP), and PSIS. This study was performed in a single centre on a sample of 42 children out of a total of 80 CHP patients, with a chronological age range between 5 days and 9.5 years from a database analysed over a period of 26 years. The study included 26/42 (62%) with CPHD and 16/42 (38%) with IGHD. The analysis of perinatal variables showed a natural delivery in 52% (11/21) of CPHD vs 13% (2/15) of IGHD. Four patients, two with CPHD and two IGHD had breech and transverse presentation respectively. All of them were resolved by caesarean section. The perinatal histories showed hypoglycaemia (61% CPHD vs 19% IGHD, P=.0105), jaundice (38% CPHDvs25% IGHD), micropenis (75%CPHD), hypoglycaemic seizures (75% CPHD), and cholestasis (19% CPHD). The mean CA of consulting for CPHD patients was 2.1 years, 30% in neonatal period and 70% before 2 years. The mean chronical age (CA) was 3.6 years in IGHD patients, with 44% of them less than 2 years. MRI showed that 81% of CPHD patients had absence of pituitary stalk vs 19% with thin pituitary stalk (P=.0001); Patients with IGHD presented 56% absence of pituitary stalk vs 44% with thin pituitary stalk (P=.5067). All (100%) of the patients diagnosed in the neonatal stage had absent pituitary stalk. The characterisation of GH deficient patients by presence and type of hypothalamic-pituitary imaging abnormality provides valuable information as a predictor of phenotypic severity, treatment response, and the potential to develop additional hormonal deficiencies. We conclude that demonstrating PSIS in children with HPC provides valuable information as a predictor of phenotypic severity, presence of MPHD, and response to treatment. The low frequency of potentially dysfunctional positional obstetric history, as part of the pathophysiological mechanisms responsible for PSIS, would indicate the need to analyse the importance of possible genetic and epigenetic factors involved. Early diagnosis of HPC should be suspected in the presence of clinical signs and symptoms, such as hypoglycaemia, cholestasis, micropenis, and associated facial midline defects. MRI should be part of complementary studies in patients with this diagnostic suspicion, especially at an early age. Late recognition of this entity may increase morbidity and mortality with potential permanent deleterious effects.

Acute Toxicity of the Insecticide Cypermethrin (CYPERMON ® 20 EC) on Four Species of Colombian Anurans / Toxicidad aguda del insecticida cipermetrina (cypermon ® 20 ec) en cuatro especies de anuros colombianos

RESUMEN El insecticida cipermetrina (CY) es usado en la agricultura para el control de plagas; sin embargo, por su acción neurotóxica puede afectar organismos no blanco como los anuros. El objetivo del trabajo fue determinar la toxicidad (concentración letal media: CL50, y algunos efectos subletales: retrasos en el desarrollo, capacidad de natación y longitud total de las larvas) del insecticida CY (Cypermon® 20EC) expuesto durante 96 horas en embriones y renacuajos de cuatro especies de anuros bajo pruebas de laboratorio y microcosmos. Los embriones de Rhinella humboldti fueron los más sensibles en condiciones de laboratorio (CL50= 6,27 mg/L) y Boana xerophylla en microcosmos (CL50= 88,32 mg/ha), mientras que los de Engystomops pustulosus fueron los más resistentes (laboratorio: CL50= 1 1,80 mg/L; microcosmos: CL50= 1 12,37 mg/ha). Rhinella marina mostró una sensibilidad intermedia. En los renacuajos no fue posible calcular los valores CL50 debido a la alta mortalidad registrada en las concentraciones experimentales en laboratorio y microcosmos, las cuales fueron 40 y 122 veces menores al valor de aplicación del insecticida (500 mg/L y 1,52 mg/ha, respectivamente). Por otra parte, se encontró una reducción significante en la longitud total y la capacidad de natación de las larvas obtenidas de los embriones expuestos a la CY, pero no en el tiempo de desarrollo. En conclusión, la exposición a la cipermetrina provocó una letalidad alta en los renacuajos y efectos subletales en estadíos tempranos del desarrollo, por lo que a las concentraciones recomendadas de aplicación, este insecticida es tóxico para las especies de estudio.

ABSTRACT The cypermethrin (CY) insecticide is used in agriculture for the control of pests; however, due to its neurological action can affect non-target organisms such as anurans. The aim of this study was to evaluate the toxicity (median lethal concentration: LC50, and some sublethal effects: developmental delays, swimming performance and total length of larvae) of the insecticide CY (Cypermon® 20EC) exposed during 96 hours to embryos and tadpoles of four anuran species under laboratory and microcosm tests. The embryos of Rhinella humboldti were the most sensitive in laboratory conditions (LC50= 6.27 mg/L) and Boana xerophylla in microcosms (LC50= 88.32 mg/ha), whereas Engystomops pustulosus (laboratory: LC50= 1 1.80 mg/L, microcosms: LC50= 1 12.37 mg/ha) was the most resistant species. Rhinella marina showed an intermediate sensitivity. It was not possible to calculate the tadpole LC50 values due to the high mortality recorded in the experimental concentrations, both in laboratory and microcosms, which were 40 and 122 times lower than the value suggested for the application of the insecticide in field (500 mg/L and 1.52 mg/ha, respectively). On the other hand, it was found a significant decreasing in the total length and swimming performance of the larvae obtained from the embryos exposed to CY, but not in the embryonic developmental time. In conclusion, the exposure to the cypermethrin produced a high tadpole lethality and sublethal effects on developing embryos; therefore, under the recommended concentrations of field exposition, this insecticide is toxic for the study species.

Determinación de factores de adhesión asociados a la formación de biopelícula en aislamientos clínicos de Staphylococcus aureus y Staphylococcus epidermidis / Determination of adhesion factors associated with biofilm formation in clinical isolates of Staphylococcus aureus and Staphylococcus epidermidis

Resumen Los factores de adhesión son determinantes de virulencia que se expresan en microorganismos que tienen la capacidad de formar biopelícula, contribuyendo a la gravedad de infecciones intrahospitalarias. Dentro de estos componentes de la superficie microbiana que reconocen moléculas de adhesión de matriz conocidas como MSCRAMMs, se incluyen el factor de unión a fibronectina A y B, (FnbA y B) factor de aglutinación A y B (ClfA y B) y factor de unión a fibrinógeno (Fib), que se han descrito en Staphylococcus aureus y reaccionan con proteínas de la matriz extracelular humana. El objetivo de este estudio fue determinar la presencia de estos factores de adhesión relacionados con la formación de biopelicula en Staphylococcus. Método. Se caracterizaron fenotípica y genotípicamente 30 aislamientos clínicos de Staphylococcus aureus y Staphylococcus epidermidis, provenientes de pacientes inmunocomprometidos en tres instituciones hospitalarias de Bogotá. La producción de Biopelícula se determinó mediante Rojo Congo y Cristal violeta y mediante PCR convencional y múltiplex se amplificaron los genes FnbA y B, ClfA y B y Fib, así como los genes del operón ica ADBC. Resultados. Todos los aislamientos clínicos fueron positivos genotípica y fenotípicamente para la producción de Biopelícula, encontrándose la presencia del operón completo en el 88.9%, los factores ClfA y ClfB en un 70%; Fib en un 60%, FnbB en un 23% y FnbA en el 17%. Conclusiones. En este estudio se evidenció la presencia de estos factores de virulencia en S. epidermidis, los cuáles hasta el momento se han reportado únicamente en S.aureus. Este hallazgo es importante ya que se sugiere la relación con transferencia horizontal de genes entre estas especies, siendo el S. epidermidis un importante reservorio genético, y un importante patobionte causal de infecciones nosocomiales, asociado con dispositivos médicos.

Abstract Adhesion factors are virulence determinants that are expressed in microorganisms with the ability to form biofilms, contributing to the severity of nosocomial infections. Among these microbial surface components recognizing adhesive matrix molecules (MSCRAMMs), are fibronectin binding A and B (fnbA and B) clumping A and B (ClfA and B) and fibrinogen binding (Fib) factors. All of these have been described in Staphylococcus aureus and react with human extracellular matrix proteins. The goal of this study was to determine adhesion factors related to the biofilm formation in Staphylococcus. Method. For these purpose, 30 clinical isolates of Staphylococcus aureus and Staphylococcus epidermidis, from immunocompromised patients in three hospitals in Bogotá were characterized both, genotypically and phenotypically. The biofilm formation was determined through Congo Red and Violet Crystal and the genes FnbA, B, ClfA, B, Fib and operon ica ADBC were amplified through conventional and multiplex PCR. Results. Every clinical isolate were genotypically and phenotypically positive for the biofilm formation, being found the presence of the whole ica ADBC operon in 88,9%. The ClfA and ClfB were found by 70%; Fib 60%, fnbB 23% and 17% offnbA. Conclusions. This study proved the presence of these virulence factors in S. epidermidis, which so far have only been reported in S. aureus. This finding is important because it suggestes the relationship with horizontal gene transfer between these species, being the S. epidermidis an important genetic reservoir and a causal patobiont of nosocomial infections associated with medical devices.

Toxicidad del herbicida propanil (propanil trust® 500ec) en embriones y renacuajos de tres especies de anuros / Toxicity of the Herbicide Propanil (Propanil Trust® 500EC) to Embryos and Tadpoles of three Anuran Species

El Propanil es un herbicida empleado en el control de arvenses que puede afectar organismos no blanco como los anuros. El objetivo del trabajo fue evaluar los efectos letales (concentración letal media, CL50) y subletales (retrasos en el desarrollo, longitud total y máxima distancia de natación) del Propanil (Propanil Trust® 500EC) sobre embriones y renacuajos de tres especies de anuros bajo condiciones de laboratorio y microcosmos, que incluyen algunos componentes de campo (sedimentos y material vegetal). La especie más sensible fue Rhinella humboldti (embriones: laboratorio CL50= 9,14 mg/L y microcosmos CL50= 83,92 mg/ha; renacuajos: laboratorio CL50= 5,09 mg/L y microcosmos CL50= 44,52 mg/ha), y la más resistente Hypsiboas crepitans (embriones: laboratorio CL50= 19,58 mg/L y microcosmos CL50= 179,53 mg/ha; renacuajos: laboratorio CL50= 16,54 mg/L y microcosmos CL50= 190,72 mg/ha). Engystomops pustulosus mostró una sensibilidad intermedia. En general, en laboratorio los CL50 indicaron una letalidad alta y se encontraron cambios significantes en la longitud total y la máxima distancia de natación de los organismos expuestos al Propanil, contrario a los resultados en microcosmos, aunque el tiempo de desarrollo embrionario no mostró diferencias entre tratamientos. Al comparar los CL50 se encontró que los renacuajos fueron más sensibles al herbicida que los embriones. En conclusión, el Propanil resultó tóxico para los embriones y renacuajos en condiciones de laboratorio pero tuvo un efecto menor en microcosmos. Esto demuestra que los componentes de campo de los microcosmos y la falta de renovación de las soluciones reducen la toxicidad del Propanil en los anuros de estudio.

Propanil is an herbicide used to control weeds that can affect non-target organisms such as anurans. The aim of this study was to assess the lethal (median lethal concentration, LC50) and sublethal effects (delays in development, total length and maximum swimming distance) of Propanil (Propanil Trust® 500EC) on embryos and tadpoles of three species of anurans under laboratory and microcosm conditions, which include some field components (sediments and vegetal materials). The most sensitive species was Rhinella humboldti (embryos: laboratory LC50= 9.14 mg/L and microcosm LC50= 83.92 mg/ha; tadpoles: laboratory LC50= 5.09 mg/L and microcosm LC50= 44.52 mg/ha), whereas the most resistant was Hypsiboas crepitans (embryos: laboratory LC50= 19.58 mg/L and microcosm LC50= 179.53 mg/ha; tadpoles: laboratory LC50= 16.54 mg/L and microcosm LC50= 190.72 mg/ha). Engystomops pustulosus showed an intermediate sensitivity. In general, in laboratory the LC50 indicated a high lethality and were found significant changes in the total length and maximum swimming distance of the organisms exposed to Propanil, as opposed to the microcosm results, although the time of embryonic development did not show differences between treatments. Comparing the LC50, it was established that tadpoles were more sensitive to the herbicide than embryos. In conclusion, the Propanil was toxic to embryos and tadpoles in laboratory tests but had a low effect in microcosms. This demonstrates that the field components of the microcosms and the lack of solution renewals notably decrease the toxicity of Propanil on the anurans studied.

Extracellular vesicles secreted by highly metastatic clonal variants of osteosarcoma preferentially localize to the lungs and induce metastatic behaviour in poorly metastatic clones.

Osteosarcoma (OS) is the most common pediatric bone tumor and is associated with the emergence of pulmonary metastasis. Unfortunately, the mechanistic basis for metastasis remains unclear. Tumor-derived extracellular vesicles (EVs) have been shown to play critical roles in cell-to-cell communication and metastatic progression in other cancers, but their role in OS has not been explored. We show that EVs secreted by cells derived from a highly metastatic clonal variant of the KHOS cell line can be internalized by a poorly metastatic clonal variant of the same cell line and induce a migratory and invasive phenotype. This horizontal phenotypic transfer is unidirectional and provides evidence that metastatic potential may arise via interclonal co-operation. Proteomic analysis of the EVs secreted by highly metastatic OS clonal variants results in the identification of a number of proteins and G-protein coupled receptor signaling events as potential drivers of OS metastasis and novel therapeutic targets. Finally, multiphoton microscopy with fluorescence lifetime imaging in vivo, demonstrated a preferential seeding of lung tissue by EVs derived from highly metastatic OS clonal variants. Thus, we show that EVs derived from highly metastatic clonal variants of OS may drive metastatic behaviour via interclonal co-operation and preferential colonization of the lungs.

Auranofin is a potent suppressor of osteosarcoma metastasis.

Osteosarcoma (OS) accounts for 56% of malignant bone cancers in children and adolescents. Patients with localized disease rarely develop metastasis; however, pulmonary metastasis occurs in approximately 50% of patients and leads to a 5-year survival rate of only 10-20%. Therefore, identifying the genes and pathways involved in metastasis, as new therapeutic targets, is crucial to improve long-term survival of OS patients. Novel markers that define metastatic OS were identified using comparative transcriptomic analyses of two highly metastatic (C1 and C6) and two poorly metastatic clonal variants (C4 and C5) isolated from the metastatic OS cell line, KHOS. Using this approach, we determined that the metastatic phenotype correlated with overexpression of thioredoxin reductase 2 (TXNRD2) or vascular endothelial growth factor (VEGF). Validation in patient biopsies confirmed TXNRD2 and VEGF targets were highly expressed in 29-42% of metastatic OS patient biopsies, with no detectable expression in non-malignant bone or samples from OS patients with localised disease. Auranofin (AF) was used to selectively target and inhibit thioredoxin reductase (TrxR). At low doses, AF was able to inhibit TrxR activity without a significant effect on cell viability whereas at higher doses, AF could induce ROS-dependent apoptosis. AF treatment, in vivo, significantly reduced the development of pulmonary metastasis and we provide evidence that this effect may be due to an AF-dependent increase in cellular ROS. Thus, TXNRD2 may represent a novel druggable target that could be deployed to reduce the development of fatal pulmonary metastases in patients with OS.

Desigualdades en la mortalidad por enfermedades cardiovasculares en los municipios del Eje Cafetero, 2009-2011 / Inequalities in mortality by cardiovascular diseases in the Colombian Coffee Growing Region, 2009-2011

Introducción. El impacto de la mortalidad por enfermedades cardiovasculares requiere medir la relación entre las condiciones socioeconómicas locales y estas causas de muerte. Objetivo. Determinar la desigualdad en la mortalidad por enfermedades cardiovasculares en los municipios del Eje Cafetero (2009-2011). Materiales y métodos. Se hizo un estudio ecológico en el que se comparó la mortalidad por enfermedades cardiovasculares (hipertensivas, isquémicas, cerebrovasculares) en los municipios con base en su situación económica. Los datos de mortalidad y el índice de necesidades básicas insatisfechas se obtuvieron de las estadísticas vitales del Departamento Administrativo Nacional de Estadística (DANE), en tanto que el producto interno bruto municipal per cápita se calculó para el estudio. Los índices de desigualdad empleados se calcularon por rangos y en modelos de regresión, así como con los índices de concentración, y de Theil, utilizando el programa Epidat 3.1. Resultados. El riesgo de morir por enfermedad isquémica e hipertensiva resultó mayor en los municipios con el mayor índice de necesidades básicas insatisfechas. La mortalidad por enfermedad hipertensiva también tendió a concentrarse en dichos municipios. Se encontraron más muertes por enfermedad hipertensiva en los municipios con menor producto interno bruto per cápita en 2009 y 2010, y por enfermedad isquémica, en 2010 y 2011. No obstante, este indicador no mide la brecha entre las comunidades pobres. Conclusiones. Se carece de indicadores de desigualdad desagregados a nivel de municipio. Los sugeridos con este propósito se calculan para el nivel nacional y departamental, lo que no favorece la caracterización de las desigualdades sociales en salud a nivel territorial.

Introduction: The impact of mortality from cardiovascular diseases requires the measurement of the relationship between the local socioeconomic conditions and these death causes. Objective: To determine the inequality in mortality from cardiovascular diseases in the municipalities of the Colombian Coffee Growing Region (2009-2011). Materials and methods: We conducted an ecological study to compare the mortality from cardiovascular diseases (hypertensive, ischemic, cerebrovascular) in municipalities and their economic situation. Mortality rates and the index of unsatisfied basic needs were obtained from the Colombian Departamento Administrativo Nacional de Estadística (DANE) vital statistics, while the municipal gross domestic product per capita was estimated for this study. The inequality indices were calculated using regression models, and concentration and Theil indices with Epidat 3.1. Results: The death risk resulting from ischemic or hypertensive diseases was greater in those municipalities with a higher index of unsatisfied basic needs. Mortality due to hypertensive disease tended to concentrate in municipalities with a higher level of unsatisfied basic needs. The municipalities with a lower gross domestic product showed a higher rate of deaths due to hypertensive disease in years 2009 and 2010, and due to ischemic disease in years 2010 and 2011. Nevertheless, this indicator does not measure the gap existing among poor communities. Conclusions: Disaggregated inequality indicators at municipal level are lacking. Suggested indicators are estimated only for country and provincial levels and they do not favor the characterization of health social inequalities at territorial level.