GC-NPD analysis of topiramate in capillary dried plasma: assessing chemical dependency pharmacotherapy.

Aim: This study aimed to develop and validate a GC-NPD method for quantifying topiramate (TPM) in capillary dried plasma spots (DPS).Materials & methods: Extraction involved three 6 mm DPS with albumin 0.1%, alkaline liquid extraction with tert-Butyl methyl ether and TMAH methylation. The method was validated and applied to 15 paired samples of capillary DPS and venous plasma from chemical dependency patients.Results: The method was linear from 1 to 50 µg/ml (r >0.99), precise (CV% 3.62-8.29%) accurate (98.1-107.7%). TPM stability was confirmed in DPS stored at 4, 23 and 45°C for 21 days. DPS TPM measurements were highly correlated plasma concentrations (rs = 0.96), representing on average 102% of the venous plasma measurements.Conclusion: The method was fully validated, demonstrating potential for clinical application.

[Box: see text].

Determinación de la microdureza del esmalte en lesiones de manchas blancas tratadas con resinas infiltrantes / Determination of enamel microhardness in white spot lesions treated with infiltrating resins

Objetivo: Determinar in vitro si la microdureza superficial Knoop de las manchas blancas mejora con la infiltración de resinas de baja viscosidad en comparación con la microdureza del esmalte sano. Materiales y métodos: Se desmineralizaron 93 muestras de esmalte bovino simulando lesiones de manchas blancas, para luego ser tratadas mediante la infiltración de resinas. Se midió la microdureza Knoop de las muestras antes de desmineralizarlas, luego de este proceso y a continuación de la infiltración. Resultados: Las muestras desmineralizadas mostraron una disminución del 50% en la microdureza. La infiltración con resinas mejoró esta propiedad, aunque los valores obtenidos se encontraron muy por debajo de la microdureza del esmalte sano. El análisis estadístico, mediante análisis de varianza (ANOVA) y prueba post-hoc de Tukey, mostraron diferencias estadísticamente significativas entre los tres grupos (p<0.001). Conclusiones: El tratamiento de las lesiones de manchas blancas artificiales con resinas infiltrantes aumentó la dureza Knoop, pero sin recuperar totalmente los valores próximos a la dureza del esmalte sano. (AU)

Aim: Determine in vitro if the Knoop surface microhardness of white spots lesions improve with the infiltration of low viscosity resins and compare it with the microhardness of healthy enamel. Materials and methods: 93 samples of bovine enamel were demineralized simulating white spot lesions, and then treated by infiltration of resins. The Knoop microhardness of the samples was measured before demineralization, after this process and after infiltration. Results: Demineralized samples showed a decrease in microhardness of 50%. The infiltration with resins improved this property, although the obtained values were far below the microhardness of healthy enamel. The statistical analysis, through analysis of variance (ANOVA) and Tukey's posthoc test, showed statistically significant differences between groups (p<0.001). Conclusions: Treatment of artificial white spot enamel lesions with infiltrating resins increased Knoop hardness but failed to reach values close to the hardness of healthy enamel. (AU)

Cafe-au-lait spots with resistant hypertension are an indicator of pheochromocytoma: a rare case report.

This case report is one of the rare cases of bilateral pheochromocytoma associated with neurofibromatosis type 1. The interest lies in the clinical form in which the diagnosis was revealed. We report the case of a 38-year-old woman admitted for severe hypertension resistant to triple therapy. Clinical examination revealed Cafe-au-lait spots, which are pigmented birthmarks that appear as patches on the skin with a light to dark brown colour. More than six spots are present in an estimated 95% of people diagnosed with neurofibromatosis type 1 (NF1). Abdominal computed tomography (CT) showed bilateral adrenal tumor involvement. The diagnosis of pheochromocytoma was made by measuring urinary Vanillylmandelic acid (VMA). The evolution was favorable after the excision of the tumor, with normalization of blood pressure. In conclusion: resistant hypertension with café au lait spots may indicate pheochromocytoma, especially bilateral, suggesting an underlying genetic condition like NF1, warranting systematic screening.

En este reporte se documenta un caso raro de feocromocitoma bilateral asociado a neurofibromatosis tipo 1. El interés radica en la forma clínica en la que se reveló el diagnóstico. Presentamos el caso de una mujer de 38 años que ingresa por hipertensión arterial severa resistente a triple terapia. El examen clínico reveló manchas café con leche, que son marcas de nacimiento pigmentadas que aparecen como manchas en la piel y que son de un color marrón claro a oscuro. Se estima que el 95% de las personas diagnosticadas con neurofibromatosis tipo 1 (NF1) presentan más de seis manchas. La tomografía computarizada (TC) abdominal mostró afectación tumoral suprarrenal bilateral. El diagnóstico de feocromocitoma se realizó mediante la medición del ácido vanilmandélico (VMA) urinario. La evolución fue favorable tras la extirpación del tumor, con normalización de la presión arterial. En conclusión, la hipertensión resistente con manchas café con leche puede indicar feocromocitoma, especialmente si es bilateral, lo que sugiere una afección genética subyacente como la NF1, que justifica un tamizaje sistemático.

Automatic Auditing System for Endoscopic Exploration of the Stomach with Artificial Intelligence-Gastro UNAL: Gastroendoscopy UNit for Automatic Labeling / Sistema de auditoría automática para la exploración endoscópica del estómago con Inteligencia Artificial - Gastro UNAL: Gastroendoscopy UNit for Automatic Labeling

Abstract Introduction: Upper endoscopy is the standard method for diagnosing early-stage gastric cancer. However, according to estimates, up to 20% of tumors are not detected, and their accuracy may be affected by the variability in their performance. In Colombia, most diagnoses take place in advanced stages, which aggravates the problem. Protocols have been proposed to ensure the complete observation of areas prone to premalignant lesions to address variability. Objective: To build and validate an automatic audit system for endoscopies using artificial intelligence techniques. Methodology: In this study, 96 patients from a teaching hospital underwent video-documented endoscopies, spanning 22 stations rearranged to minimize overlaps and improve the identification of 13 key gastric regions. An advanced convolutional network was used to process the images, extracting visual characteristics, which facilitated the training of artificial intelligence in the classification of these areas. Results: the model, called Gastro UNAL, was trained and validated with images of 67 patients (70% of cases) and tested with 29 different patients (30% of cases), which reached an average sensitivity of 85,5% and a specificity of 98,8% in detecting the 13 gastric regions. Conclusions: The effectiveness of the model suggests its potential to ensure the quality and accuracy of endoscopies. This approach could confirm the regions evaluated, alerting less experienced or trained endoscopists about blind spots in the examinations, thus, increasing the quality of these procedures.

Resumen Introducción: La endoscopia digestiva alta es el método estándar para diagnosticar el cáncer gástrico en etapas tempranas. Sin embargo, su precisión puede verse afectada por la variabilidad en su realización, y se estiman hasta 20% de tumores no detectados. En Colombia, la mayoría de los diagnósticos se realizan en etapas avanzadas, lo que agrava el problema. Para abordar la variabilidad, se han propuesto protocolos con el fin de asegurar la observación completa de áreas propensas a lesiones premalignas. Objetivo: Construir y validar un sistema de auditoría automática para endoscopias usando técnicas de inteligencia artificial. Metodología: En este estudio, 96 pacientes de un hospital universitario se sometieron a endoscopias documentadas en video, abarcando 22 estaciones reorganizadas para minimizar solapamientos y mejorar la identificación de 13 regiones gástricas clave. Se utilizó una red convolucional avanzada para procesar las imágenes, extrayendo características visuales, lo que facilitó el entrenamiento de la inteligencia artificial en la clasificación de estas áreas. Resultados: El modelo, llamado Gastro UNAL, fue entrenado y validado con imágenes de 67 pacientes (70% de los casos) y probado con 29 pacientes distintos (30% de los casos), con lo que alcanzó una sensibilidad promedio del 85,5% y una especificidad del 98,8% en la detección de las 13 regiones gástricas. Conclusiones: La eficacia del modelo sugiere su potencial para asegurar la calidad y precisión de las endoscopias. Este enfoque podría confirmar las regiones evaluadas, alertando puntos ciegos en la exploración a los endoscopistas con menos experiencia o en entrenamiento, de tal forma que se aumente la calidad de estos procedimientos.

Helicobacter pylori exposure among the Awajún of the Peruvian Amazon: Prevalence and environmental, social, and biological associations.

OBJECTIVES: Helicobacter pylori (H. pylori)-a gastric bacteria affecting almost 50% of the global population and leading to ulcers and cancer in severe cases-is a growing health concern among Indigenous populations who report a high burden of reported poor general health and gastrointestinal distress. We test hypothesized associations between H. pylori exposure patterns and environmental, social, and biological conditions among a sample of 212 Indigenous Awajún adults (112 males, 100 females, ages 18-65 years) living in the northern Peruvian Amazon. MATERIALS AND METHODS: Dried blood spots were analyzed for H. pylori-specific IgG using a recently developed enzyme-linked immunosorbent assay. Resulting seropositivity rates and antibody concentrations, proxying past exposures to H. pylori were analyzed in relation to relevant environmental (toilet type, floor material, reported water quality), social (household size and education level), and biological (age, sex, BMI, blood pressure, immune and metabolic biomarkers) factors using multivariable regression analyses. RESULTS: We found near ubiquitous seropositivity for H. pylori exposure in our sample (99.1% seropositive). In the regression analyses, elevations in H. pylori antibody concentrations were significantly higher among males compared to females (ß = 0.36, p = 0.01). No associations were found with any other factors. DISCUSSION: Anthropological research in the study communities suggests that the male bias in elevations of H. pylori antibody concentrations is related to cultural and biological factors. Future research is needed to further unravel these biocultural dynamics and determine whether elevations in H. pylori antibody concentrations have clinical relevance for gastrointestinal health outcomes in this population.

Remineralizing effect of the association of nano-hydroxyapatite and fluoride in the treatment of initial lesions of the enamel: A systematic review.

OBJECTIVE: Assessed the effect of dental products containing nano-hydroxyapatite (nano-HA) + fluoride on the remineralization of white spot lesions (WSL) in vivo or in situ. METHODS: Seven databases were explored using a two-pronged approach (intervention/treatment). After screening, full-text assessment, and further exclusion, the qualitative synthesis of five studies (four clinical and one in situ) was performed. Based on the Cochrane collaboration guidelines relevant data of the studies were collected and summarized. The Cochrane risk of bias tool for randomized trials (RoB 2.0) was used to appraise the studies' methodological quality and the GRADE guidelines to assess their level of evidence. The RoB 2.0 domains were rated on their risk of bias (RoB) as low, high, or with some concerns, and an adaptation of the tool was used to the in situ study. RESULTS: The included studies assessed 151 WSL in anterior permanent teeth, on patients with varying ages. The protocol application, treatment length (7d-12 w), and control groups varied greatly between the studies making the performance of a quantitative analysis unfeasible. The general RoB of the clinical studies was classified as being of low risk (n = 2) or some concerns (n = 2). The in situ study was considered as being of low risk. The level of the evidence was moderate. Most of the studies found moderate evidence regarding the superiority of this association in clinical settings. CONCLUSION: Even with the nano-HA + fluoride promising results for the remineralization treatment of WSL, due to the restricted number of studies and types of products, its extended use cannot be recommended based on the current systematic review, especially when considering the moderate level of the evidence found. CLINICAL SIGNIFICANCE: Due to the biocompatibility and higher surface coverage of nano-HA and the remineralization capacity of fluoride formulations, the association of these elements to remineralize WSL has been positively reported. After the collection and qualitative appraise of the data, the clinical evidence of the use of these dental products is promising but limited.

Xerophthalmia secondary to bowel malabsorption after Roux-en-Y gastric bypass.

Introduction: Vitamin A is a fat-soluble vitamin, obtained through diet. Vitamin A deficiency is the leading cause of preventable blindness in children in developing countries due to impaired intake (Phanachet et al. 2018). Nevertheless, it is uncommon in the developed world where malabsorption takes a prominent role. Case description: A fifty-one-year-old female presented complaining of foreign body sensation, pain, tearing, fluctuating visual acuity, nyctalopia, diarrhea, polyphagia and weight loss. She had history of Roux-en-Y gastro-jejunal bypass, Lynch syndrome and right hemicolectomy with ileo-colonic anastomosis, she also referred to an additional unspecified bowel resection. In the ophthalmologic examination, best corrected visual acuity was 20/30, intraocular pressure was 11 mmHg in both eyes. Anterior segment biomicroscopy revealed a dry and thickened conjunctiva with wrinkles, multiple grey-white small, round, confluent, foamy lesions in the interpalpebral conjunctiva of both eyes, compatible with Bitot's spots, and superficial punctate keratitis. Discussion: The rise of bariatric surgery, inflammatory bowel disease and end stage liver disease has led to an increase in cases of malabsorption syndrome and nutrient deficiencies in the developed world. Retinoids are essential for corneal and conjunctival epithelial cells differentiation and its deficiency is associated with a wide spectrum of ocular surface manifestations known as xerophthalmia. In this case, a gastric bypass and another unspecified bowel resection should raise the suspicion of malabsorption and nutrient deficiencies. In our patient, the diagnosis was made early and appropriate treatment was implemented before irreversible damage arose, however, vitamin A deficiency can be easily overlooked. Conclusion: In patients with xerophthalmia, interrogation should include previous history of gastrointestinal surgery, especially since bariatric surgery has become a popular technique. This is, to our knowledge, the first case report of xerophthalmia in a patient with Lynch syndrome.

Características clínico-radiológicas bucofaciales del síndrome de McCune-Albright en un adulto: reporte de un caso / Orofacial clinical-radiological features of McCune-Albright syndrome in an adult: a case report / Características clínico-radiológicas orofaciais da síndrome de McCune-Albright num adulto: relato de um caso

RESUMEN La displasia fibrosa es una enfermedad ósea benigna y progresiva, de base genética que puede afectar a uno o varios huesos. Su alta incidencia en la edad pediátrica sirvió de base para que anteriormente fuera considerada exclusiva de esta etapa; sin embargo, a la fecha, existen reportes que evidencian su desarrollo y crecimiento posterior en la segunda década de vida. Cuando está asociada a desequilibrios endocrinos y manchas café con leche, forma parte del síndrome de McCune-Albright. El presente reporte de caso muestra las características clínicas y radiológicas de la región bucofacial de un paciente adulto portador del síndrome de McCune-Albright. Las lesiones en los maxilares afectan la morfología y causan disfunción. A nivel dentario, los cambios producidos se traducen en maloclusión y defectos estructurales. En los estudios radiológicos se observaron cambios con densidad mixta y el patrón de vidrio esmerilado.

ABSTRACT Fibrous dysplasia is a benign and progressive bone disease of genetic basis that can affect one or several bones. Its high incidence in the pediatric age served as a basis for it to be previously considered exclusive to this stage; however, there are reports to date that show its development and later growth in the second decade of life. When associated with endocrine imbalances and café-au-lait spots, it is part of the McCune-Albright syndrome. The present case report shows the clinical and radiologic features of the oral-facial region of an adult patient with McCune-Albright syndrome. Lesions in the jaws affect the morphology and cause dysfunction. At the dental level, the changes produced result in malocclusion and structural defects. Radiological studies showed changes with mixed density and ground glass pattern.

RESUMO A displasia fibrosa é uma doença óssea benigna, progressiva e de base genética que pode afetar um ou mais ossos. A sua elevada incidência no grupo etário pediátrico foi a base para que anteriormente fosse considerada exclusiva desta fase; no entanto, existem relatos até à data do seu desenvolvimento e crescimento mais tardio na segunda década de vida. Quando associada a desequilíbrios endócrinos e manchas café-com-leite, faz parte da síndrome de McCune-Albright. Este relato de caso mostra as características clínicas e radiológicas da região bucofacial de um paciente adulto com síndrome de McCune-Albright. As lesões nos maxilares afetam a morfologia e provocam disfunções. A nível dentário, as alterações produzidas resultam em má oclusão e defeitos estruturais. Nos estudos radiológicos, foram observadas alterações com densidade mista e padrão de vidro despolido.

Seroprevalence of Vibrio cholerae in Adults, Haiti, 2017.

In Haiti in 2017, the prevalence of serum vibriocidal antibody titers against Vibrio cholerae serogroup O1 among adults was 12.4% in Cerca-la-Source and 9.54% in Mirebalais, suggesting a high recent prevalence of infection. Improved surveillance programs to monitor cholera and guide public health interventions in Haiti are necessary.

Standardization and validation of a novel UPLC-MS/MS method to quantify first line anti-tuberculosis drugs in plasma and dried blood spots.

Tuberculosis (TB) is a high-burden infectious disease with high prevalence and mortality rates. The first-line anti-TB drugs include isoniazid (INH), rifampicin (RMP), pyrazinamide (PZA), and ethambutol (EMB). At present, the standard method of blood sampling for therapeutic drug monitoring (TDM) analysis is venipuncture. Dried blood spots (DBS) are a minimally invasive method for collecting small quantities of whole blood from fingertips. The aim of the current study was to develop an ultrahigh-performance liquid chromatography technique coupled to tandem mass spectrometry (UPLC-MS/MS) for simultaneous quantification of the first-line anti-TB drugs in human plasma and DBS as a sampling alternative. The separation and detection conditions were optimized to quantify INH, RMP, PZA, and EMB in both matrices in an ACQUITY UPLC H Class system coupled to a XEVO TQD detector. Chromatographic separation was performed through an Acquity HSS T3 column (2.1 × 100 mm, 1.8 µm) with 0.1% formic acid in water and acetonitrile as the mobile phase. The total run time was 7 min for both methods, with retention time in plasma of 0.85, 1.22, 3.16, and 4.04 min and 0.74, 0.87, 0.97, and 4.16 min for EMB, INH, PZA, and RMP in DBS, respectively. The bioanalytical methods developed were proved selective, linear, precise, and accurate (inter- and intra-assay); the matrix effect was demonstrated to be within the established limits. Short- and long-term stability, freeze-thaw cycles for plasma, and short-term stability for DBS were established. A total of 15 patients with 46 ± 17 (mean ± SD) years old were included, and anti-TB drug concentrations were quantified on plasma and DBS as proof of concept. Based on RMP and INH plasma concentrations (Cp), and Bayesian estimation of individual pharmacokinetic parameters, a dose adjustment was necessary for 93% of patients. The slopes of the correlation lines between plasma and DBS concentrations of RMP, EMB, INH, and PZA were 0.5321, 0.8125, 0.5680, and 0.6791, respectively. Finally, significant correlations (p < 0.05) were observed between DBS and plasma concentrations for RMP (r2 = 0.6961), EMB (r2 = 0.4369), INH (r2 = 0.8675) and PZA (r2 = 0.7363). A simple, fast, and reliable UPLC-MS/MS method was developed to quantify first-line anti-TB drugs in plasma and DBS, which provides an easy sampling and storage to be applied as a new strategy for TDM in patients with TB.

Population-Based Serologic Survey of Vibrio cholerae Antibody Titers before Cholera Outbreak, Haiti, 2022.

A Vibrio cholerae O1 outbreak emerged in Haiti in October 2022 after years of cholera absence. In samples from a 2021 serosurvey, we found lower circulating antibodies against V. cholerae lipopolysaccharide in children <5 years of age and no vibriocidal antibodies, suggesting high susceptibility to cholera, especially among young children.

First Report of Pseudocercospora paraguayensis Causing Leaf Spots on Bixa orellana from Chinese Mainland.

Bixa orellana L. is a traditional Chinese medicine. In December 2019, a leaf spot disease was observed on B. orellana from a field in Zhanjiang (21°18'12''N, 110°17'22''E), China. Disease incidence was around 85% (n = 100 investigated plants from about 30 hectares). Initial leaf spots were circular, and the center of the lesions was grayish-white with a purple black border. The coalescence of individual spots eventually led to leaf wilt. Ten symptomatic leaves from 10 plants were sampled. The margins of the samples were cut into 2 mm × 2 mm pieces, and the surfaces were disinfected with 75% ethanol for 30 sec, and 2% sodium hypochlorite for 60 sec. The the samples were then rinsed three times in sterile water, plated on potato dextrose agar (PDA), and incubated at 28 °C. Pure cultures were obtained by transferring the hyphal tips to new PDA plates. Three representative isolates (BOPP-1, BOPP-2, and BOPP-3) were used for further study. The colonies of isolates on PDA were dark olive green with off-white aerial mycelia after 7 days at 28 °C. Conidia were solitary, smooth to verrucous, olive to light brown, slightly curved, narrowly obclavate, apex obtuse, base obconic-truncate, had 2-4 septa, and 30.4-55.5×2.0-3.5 µm in size.. These morphological characteristics showed did not differ from the description of Pseudocercospora paraguayensis (Crous et al. 1997). For molecular identification, the internal transcribed spacer (ITS) region, the translation elongation factor 1-α (TEF1) gene, and actin (ACT) gene were amplified using primer pairs ITS1/ITS4 (White et al., 1990), EF1/EF2 ( O'Donnell et al. 1998), and ACT-512F/ACT-783R (Carbone and Kohn, 1999) and sequenced from DNA extracted from the three isolates, respectively. Sequences were deposited in GenBank under accession no. MZ363823-MZ363825 (ITS), MZ614954-MZ614956 (TEF1), and MZ614951-MZ614953 (ACT). A phylogenetic tree was generated on the basis of the concatenated data from the sequences of ITS, TEF1, and ACT that the three isolates were nested within the clade containing the type specimen of P. paraguayensis (CBS 111286) but not within P. bixae (the type specimen CPC 25244). Pathogenicity was tested through in vivo experiments. Inoculation and control seedlings (n = 5, 1-month-old) were sprayed with a spore suspension (1 × 105 per ml) of P. paraguayensis and sterile distilled water (control), respectively, until run-off (Fang. 1998). The plants were grown in pots in a greenhouse at 28°C, with at approximately 80% RH. The test was performed three times. Symptoms similar to those in the field were observed on the inoculated plants after two weeks. The control plants remained healthy. The fungus was re-isolated from the infected leaves and confirmed as the same isolates by morphological and comparison of ITS sequences with 100% identical to those of isolates. No original fungi were isolated from the control plants. A previous study reported that P. paraguayensis caused leaf spots on pistachio and eucalypts, and the fungus causing the leaf spots of B. orellana was redescribed as P. bixae (Crous et al. 2019). However, multilocus phylogenetic analyses differentiated P. paraguayensis from P. bixae. In the present study, P. paraguayensis was distinguished from P. bixae due to the absence of catenulate conidia and the presence of finely verruculose conidia (Crous et al. 2013). P. eucalypti as a synonyms was reported in Taiwan (www.MycoBank.org). The current study is the first to report P. paraguayensis causing leaf spots on B. orellana from Chinese Mainland. This finding will help to provide a scientific basis for the disease detection.

Birefringence analysis of collagen supraorganization in cat corneas with tropical keratopathy.

OBJECTIVE: To evaluate the birefringent properties of the cornea and examine the supraorganizational aspects of collagen fibers in cats with tropical keratopathy. PROCEDURE: In this study, 10-micrometer-thick sections of corneal tissue from cats with tropical keratopathy were examined, both in the opaque and transparent areas of the anterior stroma. Control samples were obtained from healthy cat corneas. Polarized light microscopy was employed to evaluate the birefringent properties using two distinct methods. The first method involved measuring the optical retardation associated with corneal birefringence, while the second method assessed the alignment/waviness of the birefringent collagen fibers. Differences were significant when p < .05. RESULTS: Tropical keratopathy resulted in a significant rise (p < .05) in optical retardation in both opaque and transparent regions of the cat cornea. In the anterior stroma, both the opaque zones and transparent tissue exhibited a higher degree of collagen fiber packing than the control corneas. However, no significant differences (p > .05) in alignment were observed between the transparent tissue of the diseased cornea and the healthy corneas. CONCLUSION: Supraorganizational changes in collagen fiber packing are not restricted to lesion zones in cat corneas affected by tropical keratopathy. Such alterations also occur in the corneal tissue of the anterior stroma adjoining the lesions. Therefore, it is plausible that the transparent tissue of the anterior stroma in corneas affected by the disease may have functional abnormalities, despite its macroscopic healthy appearance. Additional investigations are required to clarify the implications of these potential defects and their conceivable contribution to tropical keratopathy.

Dried blood spot sampling for therapeutic drug monitoring: challenges and opportunities.

INTRODUCTION: The use of dried blood spots (DBS) has gained interest in the field of therapeutic drug monitoring (TDM) due to its potential advantages, such as minimally invasive capillary blood collection, potential stabilization of drugs and metabolites at room or high temperatures, and lower biohazard, allowing for inexpensive storage and transportation. However, there are several drawbacks to the clinical use of DBS in TDM, mostly related to hematocrit (Hct) effects, differences between venous and capillary blood concentrations, among others, that must be evaluated during analytical and clinical method validation. AREA COVERED: This review focuses on the most recent publications on the applications of DBS sampling for TDM (2016-2022), with a special focus on the challenges presented by this alternative sampling strategy, as well as the opportunities for clinical applications. Real-life studies presenting clinical applications were reviewed. EXPERT OPINION: With the availability of method development and validation guidelines for DBS-based methods in TDM, higher levels of assay validation standardization have been achieved, expanding the clinical applications of DBS sampling in patient care. New sampling devices that overcome the limitations of classical DBS, such as the Hct effects, will further encourage the use of DBS in routine TDM.

Aneurysms involving the coronary arteries in a neonate with neurofibromatosis 1.

Aneurysmal coronary artery disease (ACAD) has been reported rarely in patients with neurofibromatosis type 1 (NF1), mostly in adults. We report on a female newborn affected by NF1 with ACAD disclosed during investigation for an abnormal prenatal ultrasound along with a review of the previously reported cases. The proposita had multiple café-au-lait spots and had no cardiac symptoms. Echocardiography, and cardiac computed tomography angiography confirmed aneurysms on the left coronary artery, left anterior descending coronary artery, and of the sinus of Valsalva. Molecular analysis detected the pathogenic variant NM_001042492.3(NF1):c.3943C>T (p.Gln1315*). Literature findings on ACAD in NF1 indicated that this mostly occurs in males, showing predilection for the development of aneurysms at the left anterior descending coronary artery, and manifesting predominantly as acute myocardial infarction, inclusively in teenagers, though it may be also asymptomatic as in our case. This report documents the first case of ACAD in a patient with NF1 diagnosed at birth, emphasizing that its early diagnosis is essential to prevent potential life-threatening events attributable directly to coronary lesions.

Familial Hypercholesterolemia Biomarker Distribution in Dried Blood Spots.

OBJECTIVE: To evaluate distribution profiles of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and apolipoprotein B (apoB) as candidate markers of familial hypercholesterolemia in newborns, taking into consideration potential confounding factors, such as gestational age, birth weight, sex, and race. STUDY DESIGN: TC, LDL-C, and apoB were measured from 10 000 residual deidentified newborn dried blood spot cards. Concentrations for each biomarker were reported as multiples of the median, with emphasis on describing the 99th percentile values based on birth weight, gestational age, sex, and race. Seasonal variation of biomarkers was also explored. RESULTS: LDL-C and apoB had distribution curves with tails showing extreme elevation, whereas the distribution of TC was less elevated and had the smallest range. Neonates born at early gestational age and low birth weight had significantly greater 99th percentile of multiples of the median values for apoB but not TC or LDL-C. Differences in biomarker concentration based on sex and race were minimal. All biomarkers showed greatest concentrations in the winter as compared with summer months. CONCLUSIONS: LDL-C and apoB had distribution curves supporting candidacy for neonatal familial hypercholesterolemia screening. Future studies are needed to correlate newborn screening results with molecular testing to validate these 2 biomarkers, along with measured cholesterol levels later in childhood.

Performance of the procedure for ultra-rapid extraction and loop-mediated isothermal amplification (PURE-LAMP) method to detect malaria in Haiti.

BACKGROUND: Malaria continues to cause burden in various parts of the world. Haiti, a Caribbean country, is among those aiming to eliminate malaria within a few years. Two surveys were conducted in Haiti during which we aimed to evaluate the performance of the simple and rapid procedure for ultra-rapid extraction-loop-mediated isothermal amplification (PURE-LAMP) method with dried blood spots as an alternative diagnostic method for malaria in the context of low to very low rates of transmission. METHODS: Febrile and afebrile people were recruited from three administrative divisions within Haiti: Nippes, Sud and Grand'Anse, during the summers of 2017 (early August to early September) and 2018 (late July to late August). Their blood samples were tested by microscopy, rapid diagnostic tests (RDT), PURE-LAMP and nested PCR to detect Plasmodium infection. Sensitivity, specificity, positive and negative predictive values and kappa statistics were estimated with the nested PCR results as the gold standard. RESULTS: Among 1074 samples analyzed, a positive rate of 8.3% was calculated based on the nested PCR results. Among febrile participants, the rates in 2017 and 2018 were 14.6% and 1.4%, respectively. Three positives were detected among 172 afebrile participants in 2018 by PURE-LAMP and nested PCR, and all three were from the same locality. There was no afebrile participants recruited in 2017. The PURE-LAMP, RDT and microscopy had respective sensitivities of 100%, 85.4% and 49.4%. All of the testing methods had specificities over 99%. CONCLUSIONS: This study confirmed the high performance of the PURE-LAMP method to detect Plasmodium infection with dried blood spots and recommends its use in targeted mass screening and treatment activities in low endemic areas of malaria.

Dried Blood Spot Sampling to Assess Rifampicin Exposure and Treatment Outcomes among Native and Non-Native Tuberculosis Patients in Paraguay: An Exploratory Study.

The aim of this study was to evaluate the difference in drug exposure of rifampicin in native versus non-native Paraguayan populations using dried blood spots (DBS) samples collected utilizing a limited sampling strategy. This was a prospective pharmacokinetic study that enrolled hospitalized tuberculosis (TB) patients from both native and non-native populations receiving oral rifampicin 10 mg/kg once-daily dosing. Steady-state DBS samples were collected at 2, 4, and 6 h after intake of rifampicin. The area under the time concentration curve 0-24 h (AUC0-24) was calculated using a Bayesian population PK model. Rifampicin AUC0-24 < 38.7 mg*h/L was considered as low. The probability of target attainment (PTA) was calculated using AUC0-24/MIC > 271 as a target and estimated MIC values of 0.125 and 0.25 mg/L. In total, 50 patients were included. Native patients (n = 30) showed comparable drug exposure to the non-natives (n = 20), median AUC0-24 24.7 (17.1-29.5 IQR) and 21.6 (15.0-35.4 IQR) mg*h/L (p = 0.66), respectively. Among total patients, only 16% (n = 8) had a rifampicin AUC0-24 > 38.7 mg*h/L. Furthermore, PTA analysis showed that only 12 (24%) of the patients met a target AUC0-24 /MIC ≥ 271, assuming an MIC of 0.125 mg/L, which plummeted to 0% at a wild-type MIC of 0.25 mg/L. We successfully used DBS and limited sampling for the AUC0-24 estimation of rifampicin. Currently, our group, the EUSAT-RCS consortium, is preparing a prospective multinational, multicenter phase IIb clinical trial evaluating the safety and efficacy of high-dose rifampicin (35 mg/kg) in adult subjects using the DBS technique for AUC0-24 estimation.