Plasma cell-free DNA as a monitoring tool for high-risk pregnancies associated with antiphospholipid syndrome.

INTRODUCTION: Despite thromboprophylaxis, women with antiphospholipid syndrome (APS) face high-risk pregnancies due to proinflammatory and prothrombotic states. This highlights the need for new monitoring and prognostic tools. Recent insights into the pathophysiological role of neutrophil activation and extracellular trap (NET) formation in this syndrome led to the exploration of plasma cell-free DNA (cfDNA), a derivative of NETosis, as a promising biomarker. MATERIALS AND METHODS: cfDNA was isolated and quantified from plasma samples of healthy pregnant women (control group, HC) and women with APS (APS group). We assessed the physiological variability of cfDNA across the three trimesters in HC. Levels of cfDNA were compared between APS and HC by gestational trimester. ROC curve analysis was performed to evaluate the efficacy of cfDNA levels for classifying APS patients. Furthermore, cfDNA levels in pregnant women with APS with obstetric complications were compared to those from uncomplicated pregnancies. RESULTS: Among HC, cfDNA significantly increased in the third trimester compared to the first and second. Elevated cfDNA levels in APS compared to HC were observed in the first and second trimesters. First-trimester cfDNA levels demonstrated the highest classification ability to discriminate between APS and HC patients (AUC: 0.906). Among APS, those with complicated pregnancies (fetal growth restriction, preeclampsia, placenta accreta) exhibited significantly elevated cfDNA levels in the second trimester. CONCLUSIONS: Elevated levels of cfDNA in pregnant women with APS, particularly among those with obstetric complications, supports further investigation into the potential of cfDNA as a valuable tool in the obstetric management of women with APS.

A Reliable Method for Quantifying Plasma Cell-Free DNA Using an Internal Standard Strategy: Evaluation in a Cohort of Non-Pregnant and Pregnant Women.

The use of plasma cell-free DNA (cfDNA) as a useful biomarker in obstetric clinical practice has been delayed due to the lack of reliable quantification protocols. We developed a protocol to quantify plasma cfDNA using an internal standard strategy to overcome difficulties posed by low levels and high fragmentation of cfDNA. cfDNA was isolated from plasma samples of non-pregnant (NP, n = 26) and pregnant (P, n = 26) women using a commercial kit and several elution volumes were evaluated. qPCR parameters were optimized for cfDNA quantification, and several quantities of a recombinant standard were evaluated as internal standard. Absolute quantification was performed using a standard curve and the quality of the complete method was evaluated. cfDNA was eluted in a 50-µl volume, actin-ß (ACTB) was selected as the target gene, and qPCR parameters were optimized. The ACTB standard was constructed and 1000 copies were selected as internal standard. The standard curve showed R2 = 0.993 and E = 109.7%, and the linear dynamic range was defined between 102 and 106 ACTB copies/tube. Repeatability and reproducibility in terms of CV were 19% and up to 49.5% for ACTB copies per milliliter of plasma, respectively. The range of cfDNA levels was 428-18,851 copies/mL in NP women and 4031-2,019,363 copies/mL in P women, showing significant differences between the groups. We recommend the application of internal standard strategy for a reliable plasma cfDNA quantification. This methodology holds great potential for a future application in the obstetric field.

Niveles de Interferón Tipo I en pacientes con Lupus Eritematoso Sistémico / Type I interferon levels in patients with systemic lupus erythematosus

Introducción: El interferón (IFN) tipo I es una citoquina que juega un rol fundamental en la patogenia del Lupus Eritematoso Sistémico (LES). Diferentes niveles de esta citoquina podrían explicar la heterogeneidad de esta patología y ser útil para evaluar la actividad de la misma. Objetivos: Determinar los niveles de IFN tipo I sérico en pacientes con LES y evaluar su utilidad como biomarcador de actividad. Material y Métodos: 16 pacientes con LES (ACR 1997) y 16 controles. Métodos: Actividad de la enfermedad (SLEDAI-2K), daño orgánico (SLICC), IFN tipo I (HEK-Blue-IFNα/β), anticuerpos anti-DNAdc (Inmunofluorescencia Indirecta), anticuerpos anti-ENA (ELISA), C3-C4 (Inmunoturbidimetría). Estadística: InfoStat/Instat/MedCalc. Valores de p<0,05 fueron considerados estadísticamente significativos. Resultados: Se observó un aumento de la concentración de IFN en el grupo LES con respecto al control (p<0,05). Los pacientes con valores de IFN superiores al punto de corte, se asociaron con la presencia de anticuerpos anti-DNAdc (OR:13,33; p<0,05). Pacientes con hipocomplementemia y aquellos con puntaje de SLEDAI-2K mayor a 8 presentaron mayores niveles de IFN comparados con pacientes con complemento normal y menor puntaje de índice, respectivamente (p<0,05). Conclusiones: Estos resultados sugieren la importancia que podría tener la determinación de IFN tipo I para el monitoreo de la actividad del LES.

Introduction: Type I interferon (IFN) is a cytokine that plays a fundamental role in the pathogenesis of Systemic Lupus Erythematosus (SLE). Different levels of this cytokine could explain the heterogeneity of this pathology and be useful to evaluate its activity. Objectives: To determine the serum type I IFN levels in patients with SLE and evaluate its usefulness as a biomarker of activity. Material and Method: 16 patients with SLE (ACR 1997) and 16 controls. Methods: Disease activity (SLEDAI-2K), organ damage (SLICC), type I IFN (HEK-Blue-IFNα/β), anti-dsDNA antibodies (Indirect Immunofluorescence), anti-ENA antibodies (ELISA), C3-C4 (Immunoturbidimetry). Statistics: InfoStat/Instat/MedCalc. P values <0.05 were statistically significant. Results: An increase in IFN concentration was observed in the SLE group respect to the control (p <0.05). Patients with IFN values above the cut-off point were associated with the presence of anti-dsDNA antibodies (OR: 13.33; p<0.05). Hypocomplementemic patients and those with a SLEDAI-2K score greater than 8 had higher IFN levels compared to patients with normal complement and a lower index score, respectively (p<0.05). Conclusions: These results suggest the importance that the determination of IFN type I could have for the monitoring of SLE activity.

Efecto de la edad en la seroprevalencia de anticuerpos IgG antivirus de Hepatitis A en el personal de salud

RESUMEN Introducción: La incorporación de la vacuna de la Hepatitis A (HA) en el año 2005 y las mejoras en las condiciones sanitarias y socio-económicas disminuyeron la endemicidad de dicha enfermedad en nuestro país, pero generaron un corrimiento etario de la población vulnerable. El equipo de salud pertenece al grupo de riesgo ocupacional con recomendación de vacunación. Nuestro objetivo fue analizar el efecto de la edad sobre los niveles de anticuerpos IgG anti virus de la HA (VHA) en un grupo de trabajadores del ámbito sanitario del Hospital Córdoba. Materiales y Métodos: Se determinó IgG anti VHA por quimioluminiscencia. Criterios de exclusión: haber padecido HA y/o haber recibido la vacuna. Se realizó una breve encuesta para obtener datos epidemiológicos. Para el análisis estadístico se utilizó el test t-Student, una curva ROC y la prueba Chi-cuadrado. Valores de p<0,05 fueron considerados estadísticamente significativos. Resultados: El grupo en estudio estuvo constituido por 90 profesionales entre 25-69 años. La seroprevalencia de IgG antiVHA fue 62,2%. Se detectaron 34 personas seronegativas con una edad promedio de 37.6±10.6, que fue significativamente menor a la del grupo seropositivo 45.3±10.8 años (p=0.0014). El punto de corte etario fue 41 años (p=0,0001; ABC=0,709; S=60,7%; E=73,5%). Los menores de 41 años presentaron una asociación estadísticamente significativa con IgG anti VHA negativa, OR=3.57. Conclusiones: Los profesionales menores de 41 años se asocian a serología anti VHA negativa. Debería evaluarse la indicación de realizar control serológico y vacunar a todo el personal que no presente anticuerpos protectivos al ingreso hospitalario

ABSTRACT Introduction: The incorporation of the hepatitis A (HA) vaccine in 2005 and the improvements in sanitary and socioeconomic conditions reduced the endemicity of this disease in our country, but generated an age shift of the vulnerable population. The health personnel belongs to the occupational risk group with recommendation of vaccination. Our aim was analyze the effect of age on the levels of IgG antibodies against hepatitis A virus (HAV) in a group of health workers at the Hospital Córdoba. Materials and Methods: HAV-specific IgG antibodies was determined by chemiluminescence. Exclusion criteria: to have suffered HA and/or received HA vaccine. A brief survey was made to obtain epidemiological data. We used the t-Student test, ROC curve and Chi-square test for statistical analysis. Values of p <0.05 were considered statistically significant. Results: The study group was formed by 90 professionals between 25-69 years. The seroprevalence of HAV-specific IgG antibodies was 62.2%. 34 seronegative persons were detected with a mean age of 37.6±10.6, which was significantly lower than the mean age of the seropositive group 45.3±10.8 years (p=0.0014). The age cut-off was 41 years (p = 0.0001; ABC = 0.709; S = 60.7%; E = 73.5%). Individuals under the age of 41 had a statistically significant association with negative HAV-IgG, OR = 3.57. Conclusions: Professionals under 41 years are associated with negative HAV IgG antibodies values. The indication to perform serological control and vaccinate any professionals who do not present protective antibodies at the time of hospital admission should be evaluated.

[Relationship between midcarpal inclination angle and scaphoid kinematic]. / Relation entre l'angle d'inclinaison médiocarpienne et la cinématique du scaphoïde.

OBJECTIVE: To investigate if there is a correlation between the so-called midcarpal inclination angle and the kinematic behavior of the scaphoid. PATIENTS AND METHODS: The population studied was 60 patients with postero-anterior radiographs of the wrist in full radial and ulnar deviation. Each patient was assessed for the type of lunate by two independent observers. For each pair of radiographs the Midcarpal Inclination Angle and the Scaphoid Flexion Index (SFI) was determined. RESULTS: Twenty-three cases were classified as lunate type I, 19 cases as type II. The average midcarpal inclination angle was 55.2° (SD±6.1) for wrists with a lunate type I and 63.8° (DE±6.3) for type II (p<0.0001). There was a significant linear relationship between the midcarpal inclination angle and the Scaphoid Flexion Index (p=0.02). CONCLUSIONS: The wrists with a midcarpal inclination angle greater than 60° (type II lunate) had a scaphoid rotating according to a "columnar pattern", during radioulnar inclinations (predominant rotation along the sagittal plane), while the wrists with a lunate type I behave according to a "row pattern".

[Analysis of the composition of 2700 urinary calculi examined by infrared spectrophotometry, 1979-1989]. / Analisi della composizione di 2.700 calcoli urinari, esaminati con spettrofotometria nell'infrarosso, nel periodo 1979-1989.

Two thousand seven hundred samples, received as urinary calculi, in a Turin based laboratory, in the period between March 1979 and February 1989, were examined by infrared spectroscopy to identify components even of difficult characterization: a special procedure was used and it is described. Knowing the composition of each calculus, a comparison was made between the incidence of each compound in the first 1500 samples (1979-1984) and in the remaining 1200 ones (1984-1989). Besides an increase in calcium oxalate as main component, a marked reduction in incidence of struvite was found in the calculi belonging to the second period of observation. The possible interference in calculi composition by various therapy for renal infection is discussed.

Impaired fibrinolysis in uremia: partial and variable correction by four different dialysis regimes.

In addition to a hemorrhagic diathesis, uremia is accompanied by a clotting tendency, caused by a marked fall in fibrinolytic capacity. Measurement of lysis time of whole blood diluted with phosphate and acetate buffers and of euglobulin lysis times showed that accumulation of inhibitors is primarily responsible. These probably belong to the class of small molecules abnormally retained in uremia. Hemodialysis (HD) offers the best method of correction, mainly because of better elimination of these inhibitors. In contrast, hemofiltration (HF) and, particularly, intermittent peritoneal dialysis (IPD) are much less effective. In IPD, protein loss via the peritoneum is also responsible for a loss of fibrinolytic activators, so that fibrinolysis becomes even poorer, exposing the patient to an increased risk of vascular complications.