Association of SNP rs5069 in APOA1 with Benign Breast Diseases in a Mexican Population.

Breast cancer (BCa) is the most common type of cancer affecting women worldwide. Some histological subtypes of benign breast disease (BBD) are considered risk factors for developing BCa. Single nucleotide polymorphisms (SNPs) in the genes encoding apolipoproteins A-I (APOA1) and B (APOB) have been associated with BCa in Tunisian, Chinese, and Taiwanese populations. The objective of this pilot study is to evaluate the possible contribution of APOA1 and APOB polymorphisms to BCa and BBD in the Mexican population. We analyzed the association of 4 SNPs in genes encoding apolipoproteins: rs670 and rs5069 in the APOA1 gene, and rs693 and rs1042031 in the APOB gene, by performing PCR-RFLP with DNA extracted from the biopsy tissue of Mexican women with BCa or BBD and whole blood samples obtained from the general population (GP). Our results showed an association between the CT + TT genotypes of the SNP rs5069 and BBD (p = 0.03201). In the A-T haplotype, the frequency of the SNPs rs670 and rs5069 differed significantly between the BBD group and the GP and BCa groups (p = 0.004111; p = 0.01303). In conclusion, the SNP rs5069 is associated with BBD but not with BCa in the Mexican population.

CCR2/CCL2 and CMKLR1/RvE1 chemokines system levels are associated with insulin resistance in rheumatoid arthritis.

Rheumatoid arthritis (RA) has been associated with insulin resistance (IR). Due to an excess in storage of white adipose tissue, IR has an inflammatory process that overlaps with RA. This is performed by the activation/migration of monocytes carried out by the CCR2/CCL2 and CMKLR1/RvE1 chemokines systems. Furthermore, these can potentiate chronic inflammation which is the central axis in the immunopathogenesis of RA. We evaluated the association between the relative expression of CCR2 and CMKLR1 and the serum levels of their ligands CCL2 and RvE1, in the context of adiposity status with IR as a comorbidity in RA. We studied 138 controls and 138 RA-patients classified with and without IR. We evaluated adiposity, RA activity, IR status and immunometabolic profiles by routine methods. Insulin, CCL2 and RvE1 serum levels were determined by ELISA. Relative expression of CCR2, CMKLR1 and RPS28 as constitutive gene by SYBR green RT-qPCR and 2-ΔΔCT method. Increased measurements were observed of body adiposity and metabolic status as follows: RA with IR>control group with IR>RA without IR> control group without IR. CCR2 and CMKLR1 relative expression was increased in RA without IR versus control without IR. CCR2: 2.3- and 1.3-fold increase and CMKLR1: 3.5- and 2.7-fold increase, respectively. Whereas, CCR2 expression correlates with CMKLR1 expression (rho = 0.331) and IR status (rho = 0.497 to 0.548). CMKLR1 expression correlates with inflammation markers (rho = 0.224 to 0.418). CCL2 levels were increased in the RA groups but levels of RvE1 were increased in RA without IR. We conclude that in RA with IR, the chemokine receptors expression pattern showed a parallel increase with their respective ligands. RA and IR in conjunction with the pathological distribution of body fat mass might exacerbate chronic inflammation. These results suggest that high CCL2 levels and compensatory RvE1 levels might not be enough to resolve the inflammation by themselves.

In vivo assessment and characterization of lactic acid bacteria with probiotic profile isolated from human milk powder.

INTRODUCTION: Introduction: breast milk (MH) contains nutrients and bioactive compounds for child development, including probiotic bacteria, which contribute to intestinal maturation. This benefit accompanies the individual until adulthood. There are new methods such as spray drying that give this compound a good conservation without loss of microbiota. Objective: the aim of this study was to analyze the viability of lactic acid bacteria isolated from human milk with probiotic potential after the spray drying process, as well as to evaluate the possible adhesion in the colon of mice of the Balb/C strain after feeding them powdered human milk and a commercial formula milk. Method: we isolated and identified the presence of lactic acid bacteria with possible probiotic potential in powdered human milk using the MALDI-TOF MS technique. Powdered human milk and a commercial formula milk were fed to mice of the Bald/C strain for 14 weeks. Glucose level and weight were measured in the mice. The feces were collected to verify the presence of lactic bacteria. The mice were sacrificed and their intestines were weighed, isolating the lactic acid bacteria both from the intestines and from the feces. The strains isolated from mice fed human milk were evaluated for their probiotic potential, analyzing their ability to inhibit pathogens, resistance to pH, temperature, adhesion, and hydrophobicity. Results: the presence of Lactobacillus fermentum LH01, Lactobacillus rhamnosus LH02, Lactobacullis reuteri LH03, and Lactobacillus plantarum LH05 in powdered human milk was identified. All strains showed a possible probiotic profile due to the ability of bacteria to resist low pH, bile salts, and exposure to gastric enzymes, as well as their hydrophobicity and self-aggregation capacity, and their failure to show hemagglutination or hemolysis activity in a culture medium rich in erythrocytes. We observed that the consumption of powdered human milk prevented weight gain and constipation in mice. Conclusions: after spray drying, strains with possible probiotic potential may be preserved in human milk. The consumption of powdered human milk with probiotic bacteria prevents constipation and weight gain in mice, when compared to those fed a commercial formula milk.

INTRODUCCIÓN: Introducción: la leche materna (HM) contiene los nutrientes y compuestos bioactivos necesarios para el desarrollo infantil, incluidas bacterias probióticas, que contribuyen a la maduración intestinal. Objetivo: el objetivo de este estudio fue analizar la viabilidad de las bacterias acidolácticas aisladas de la leche humana con potencial probiótico, después del proceso de secado, así como evaluar su posible adhesión en el colón de ratones (BAlb/C) alimentados con leche humana en polvo y leche de una fórmula comercial. Método: se aislaron e identificaron mediante la técnica de Maldi-Tof-MS las bacterias acidolácticas con posible potencial probiótico en la leche humana en polvo. Se alimentó con leche humana en polvo y leche de una fórmula comercial a ratones de la cepa Bald/C durante 14 semanas. Se midieron el nivel de glucosa y el peso. Las heces se recolectaron para verificar la presencia de bacterias lácticas. Los ratones se sacrificaron y se pesaron los intestinos, aislando las bacterias lácticas tanto de los intestinos como de las heces. En las cepas aisladas de la leche humana se evaluó el potencial probiótico analizando su capacidad para inhibir patógenos, resistir distintos pH y temperaturas, adherirse y mostrar hidrofobicidad. Resultados: se identificó la presencia de Lactobacillus fermentum LH01, Lactobacillus rhamnosus LH02, Lactobacullis reuteri LH03 y L. plantarum LH05 en la leche humana en polvo. Todas las cepas mostraron resistencia a los pH bajos, a las sales biliares y a la exposición a enzimas gástricas, así como una buena hidrofobicidad y capacidad de autoagregación. Además, no presentaron actividad de hemaglutinación o hemólisis en un medio de cultivo rico en eritrocitos. Observamos que el consumo de leche humana en polvo evita en los ratones el aumento de peso y el estreñimiento. Conclusiones: después del secado por aspersión, las cepas con posible potencial probiótico pueden conservarse en la leche materna. El consumo de leche humana en polvo con bacterias probióticas evita el estreñimiento y el aumento de peso en los ratones, en comparación con los alimentados con leche de una formula comercial.

Association between nutritional risk markers and polymorphisms rs2291166 in TJP1 and VNTR (CAG)n in ATXN2 in an obese adolescent Mexican population. / Asociación entre marcadores de riesgo nutricionales con los polimorfismos rs2291166 de TJP1 y el VNTR (CAG)n de ATXN2 en población mexicana adolescente con obesidad.

OBJECTIVE: To estimate the correlation between indices of diet quality (DQIs), insulin sensitivity (QUICKI) and resistance (HOMA-IR), waist circumference (WHR) and body mass (BMI) and the alleles and genotypes of the TJP1 SNP rs2291166 and the VNTR of ATXN2 in adolescent patients. SUBJECTS AND METHOD: The study enrolled 85 subjects aged 10-20years, from the city of Tuxtepec, Oaxaca, Mexico, recruited in the period 2017-2018. DQIs, BMI, WHR, HOMA-IR, QUICKI, and diet quality index were measured. The rs2291166 polymorphism in TJP1 was determined by allele-specific PCR and the (CAG)n expansion in ATXN2 was determined by hot start PCR. PCR products were analyzed using 8% PAGE electrophoresis and silver nitrate staining. RESULTS: A correlation was found of indices DQIs, HOMA-IR, WHR and BMI with the heterozygous genotype of the TJP1 SNP rs2291166 and the long and short repeats of the ATXN2 CAG repeat in obese adolescent patients. A very strong positive correlation was seen between the TJP1 SNP and the HOMA-IR index (P<.05). A positive correlation was also found between the ATXN2 CAG repeat and the QUICKI index (P=.000) (P<.05), while the DQIs index correlated more closely with BMI and WHR. CONCLUSIONS: DQIs, TJP1 SNP rs2291166, and ATXN2 CAG repeat are determinants of obesity-related risk parameters such as BMI, WHR, QUICKI, and HOMA-IR in the adolescent population analyzed.

In-vivo assessment of the protection of ß-glucans of Pleurotus ostreatus against oxidative stress caused by acrylamide intake (part II).

INTRODUCTION: Introduction: in April 2002, the National Food Authority of Sweden published a study in which the presence of a carcinogen was reported for the first time in experimental animals, and was identified as acrylamide. Various studies have shown that the ß-glucans of Pleurotus ostreatus have diverse biological properties including antioxidant and anticancer activities. Methods: ß-glucans were obtained by alkaline-acid hydrolysis from Pleurotus ostreatus, and their content was characterized by liquid chromatography. To evaluate the effect of ß-glucans on the expression of glutathione, Balb/c mice were used, and 4 test groups were established. All groups were fed as usual, groups treated with acrylamide were administered the compound intragastrically at a concentration of 50 g/mL, and ß-glucan treatment was given at a concentration of 50 g/mL. Results: no mortality was observed after exposure to the tested dose of acrylamide; only signs of peripheral neuropathy such as hyperactivity and tremors were observed after five days of experimentation, and were maintained over 30 days after the experiment. On the other hand, an increase in lipid peroxidation levels was observed in the livers of the acrylamide-treated mice, which were lower in the mice treated with ß-glucans. Conclusions: results show that ß-glucans may act as antioxidant agents able to protect the liver against oxidative stress as caused by the intake of acrylamide.

INTRODUCCIÓN: Introducción: en abril de 2002, la Autoridad Nacional de Alimentos de Suecia publicó un estudio en el que se informó por primera vez de la presencia de un carcinógeno en animales experimentales, identificado como acrilamida. Diversos estudios han demostrado que los ß-glucanos de Pleurotus ostreatus tienen diversas propiedades biológicas, tales como actividades antioxidantes y anticancerígenas. Métodos: los ß-glucanos se obtuvieron por hidrólisis ácido-alcalina de Pleurotus ostreatus y su contenido se caracterizó por cromatografía líquida. Para evaluar el efecto de los ß-glucanos sobre la expresión de glutatión, se usaron ratones Balb/c y se establecieron 4 grupos de prueba; todos los grupos se alimentaron normalmente, en los grupos tratados con acrilamida esta se administró intragástricamente a una concentración de 50 µg/mL, y el tratamiento con ß-glucanos se dio a una concentración de 50 µg/mL. Resultados: no se observó mortalidad después de la exposición a la dosis probada de acrilamida; solo se observaron signos de neuropatía periférica, como hiperactividad y temblores, después de cinco días de experimentación, que se mantuvieron dentro de los 30 días posteriores al experimento. Por otro lado, se observó un aumento de los niveles de peroxidación lipídica en los hígados de los ratones tratados con acrilamida, que fueron más bajos en los ratones tratados con ß-glucanos. Conclusiones: los resultados muestran que los ß-glucanos podrían actuar como agentes antioxidantes y proteger el hígado contra el estrés oxidativo causado por la ingesta de acrilamida.

In vivo assessment of the protection conferred by ß-glucans from Pleurotus ostreatus against the harmful effects of acrylamide intake (Part I).

INTRODUCTION: Introduction: acrylamide is formed in food through Maillard's reaction during thermal processing, and has been shown to be neurotoxic in humans, and a possible carcinogen. Studies have shown that ß-glucans from Pleurotus ostreatus have diverse biological properties such as antioxidant and anticancer activities. Objective: the aim of this work was to evaluate the protective effect of ß-glucans from Pleurotus ostreatus against the harmful effects of acrylamide consumption in mice. Methods: ß-glucans were obtained by alkaline-acid hydrolysis of Pleurotus ostreatus, and the content was characterized by liquid chromatography. To evaluate the effect of ß-glucans on the expression of glutathione, Balb/c mice were used, and 4 test groups were established. All groups were fed normally, and the groups treated with acrylamide were administered the compound intragastrically at a concentration of 50 g/mL; ß-glucans were administered at a concentration of 50 g/mL. Results: mice exposed to acrylamide showed a marked variation in the activity of glutathione enzymes in the liver. Significant differences (p < 0.05) were only found in the expression of glutathione transferase, which was increased almost 3 times in the group treated with ß-glucans as compared with the control group, and 1.5 times as compared with the group treated with acrylamide. Conclusions: the results show that ß-glucans could act by increasing the activity of enzymes involved in xenobiotic detoxification, thus protecting the biological system against the harmful effects caused by acrylamide intake.

INTRODUCCIÓN: Introducción: la acrilamida se forma en los alimentos a través de la reacción de Maillard durante el proceso térmico, y ha demostrado ser neurotóxica en humanos y un posible carcinógeno. Algunos estudios han demostrado que los ß-glucanos de Pleurotus ostreatus tienen diversas propiedades biológicas, como actividades antioxidantes y anticancerígenas. Objetivo: el objetivo de este trabajo fue evaluar el efecto protector de los ß-glucanos de Pleurotus ostreatus contra los efectos nocivos por consumo de acrilamida en ratones (prueba in vivo). Métodos: los ß-glucanos se obtuvieron por hidrólisis ácido-alcalina de Pleurotus ostreatus y su contenido se caracterizó por cromatografía líquida. La oxidación de los lípidos se evaluó mediante el método de TBARS, y para evaluar el efecto de los ß-glucanos en la expresión de glutatión se usaron ratones Balb/c, y se establecieron 4 grupos de prueba. Todos los grupos fueron alimentados normalmente; a lo grupos tratados con acrilamida, esta se les administró intragástricamente en una concentración de 50 µg/ml, y los ß-glucanos en una concentración de 50 µg/ml. Resultados: en el presente trabajo, los ratones expuestos a acrilamida mostraron una marcada variación en la actividad de las enzimas de glutatión determinadas en el hígado. Solo se encontraron diferencias significativas (p < 0,05) en la expresión de glutatión-transferasa, que aumentó casi 3 veces en el grupo tratado con ß-glucano en comparación con el grupo de control, y 1,5 veces con respecto al grupo tratado con acrilamida. Conclusiones: los resultados muestran que los ß-glucanos podrían actuar como agentes antioxidantes que protegen el hígado contra el estrés oxidativo causado por la ingesta de acrilamida.

Downregulation of hippocampal NR2A/2B subunits related to cognitive impairment in a pristane-induced lupus BALB/c mice.

Neuropsychiatric systemic lupus erythematosus (NPSLE) is associated with learning and memory deficit. Murine model of lupus induced by pristane in BALB/c mice is an experimental model that resembles some clinical and immunological SLE pathogenesis. Nevertheless, there is no experimental evidence that relates this model to cognitive dysfunction associated with NR2A/2B relative expression. To evaluate cognitive impairment related to memory deficits in a murine model of lupus induced by pristane in BALB/c mice related to mRNA relative expression levels of NR2A/2B hippocampal subunits in short and long-term memory task at 7 and 12 weeks after LPS exposition in a behavioral test with the use of Barnes maze. A total of 54 female BALB/c mice 8-12 weeks old were included into 3 groups: 7 and 12 weeks using pristane alone (0.5 mL of pristane) by a single intraperitoneal (i.p.) injection. A control group (single i.p. injection of 0.5 mL NaCl 0.9%) and pristane plus LPS exposure using single i.p. pristane injection and LPS of E. coli O55:B5, in a dose of 3mg/kg diluted in NaCl 0.9% 16 weeks post-pristane administration. To determine cognitive dysfunction, mice were tested in a Barnes maze. Serum anti-Sm antibodies and relative expression of hippocampal NR2A/2B subunits (GAPDH as housekeeping gene) with SYBR green quantitative reverse transcription polymerase chain reaction and 2-ΔΔCT method were determined in the groups. Downregulation of hippocampal NR2A subunit was more evident than NR2B in pristane and pristane+LPS at 7 and 12 weeks of treatment and it is related to learning and memory disturbance assayed by Barnes maze. This is the first report using the murine model of lupus induced by pristane that analyzes the NMDA subunit receptors, finding a downregulation of NR2A subunit related to learning and memory disturbance being more evident when they were exposed to LPS.

Ageing influences the relationship of circulating miR-33a and miR- 33b levels with insulin resistance and adiposity.

OBJECTIVE: The identification of circulating microRNAs related to abnormal metabolic function may be useful in the context of ageing, adiposity and insulin resistance. The miR-33 a/b has been shown to control the expression of genes involved in fatty acid biosynthesis, impaired metabolism and insulin resistance. In this study, we aimed to identify differences in circulating miR-33 a/b levels according to age-related metabolic impairment and increased adiposity. METHODS: This study included 80 individuals (30.2% with obesity, 70% females) classified according to insulin resistance (Stern's criteria) and age [young (20-39 years) and senior (40-59 years)]. Body fat was evaluated using bioelectrical impedance, biochemical markers by colorimetric, enzymatic and immuno-turbidimetry methods. TaqMan measures of circulating miR-33 a and miR-33 b with quantitative reverse transcription polymerase chain reaction in serum were assessed in association with clinical outputs. RESULTS: Circulating miR-33 a and miR-33 b levels showed significant association with fatness, the lipid profile and biomarkers of impaired glucose metabolism. Both miR-33 a and miR-33 b were associated with visceral adiposity index in non-insulin resistance and insulin resistance individuals. More important, for miR-33 a circulating levels in senior group, receiver operating characteristic curve analyses showed area under the curve 0.804 ( p = 0.010; 95% confidence interval = 0.655-0.952). CONCLUSION: Ageing influenced the relationship of circulating miR-33 a and miR-33 b with insulin resistance and increased adiposity.

Evaluación clínica y ultrasonográfica de la glándula tiroides en pacientes con artritis reumatoide.

Introducción: Los pacientes con artritis reumatoide pueden desarrollar enfermedad tiroidea autoinmune (ETA), cuyo diagnóstico clínico puede ser difícil debido a que ambas comparten síntomas como artralgias, mialgias, rigidez matutina o fatiga. Objetivo: Determinar la prevalencia de ETA en pacientes con artritis reumatoide. Método: Estudio transversal que incluyó 78 pacientes con artritis reumatoide y 81 controles clínicamente sanos pareados por edad y sexo. A ambos grupos se realizó cuantificación de anticuerpos antitiroideos, pruebas de función tiroidea, ultrasonido y biopsia de glándula tiroides cuando la puntuación de Thyroid Imaging Reporting and Data System (TIRADS) fue ≥ 4. Resultados: 24.4 % de los pacientes con artritis reumatoide presentó hipotiroidismo (p = 0.003) y altos títulos de anticuerpos antitiroideos versus controles clínicamente sanos; 53 % de los ultrasonidos tiroideos resultó normal en pacientes hipotiroideos; en pacientes con artritis reumatoide positivos para anticuerpos antitiroideos se encontró perfusión incrementada en 40 %. Los casos clasificados como TIRADS 4 fueron enviados a aspiración, con resultado histopatológico benigno. Conclusiones: Se demostró el valor clínico agregado de la evaluación tiroidea en pacientes con artritis reumatoide, conforme a la prevalencia de hipotiroidismo subclínico, positividad de anticuerpos antitiroideos y anomalías en el ultrasonido independientes de la función tiroidea normal o alterada. Introduction: Patients with rheumatoid arthritis can develop autoimmune thyroid disease (ATD), the clinical diagnosis of which can be difficult because both entities share symptoms such as arthralgia, myalgia, morning stiffness or fatigue. Objective: To determine the prevalence of ATD in patients with rheumatoid arthritis. Method: Cross-sectional study that included 78 patients with rheumatoid arthritis and 81 clinically healthy controls matched by age and gender. Both groups underwent anti-thyroid antibodies quantification, thyroid function tests, thyroid ultrasound and thyroid gland biopsy when the Thyroid Imaging Reporting and Data System (TIRADS) score was ≥ 4. Results: Hypothyroidism was found in 24.4% of patients with rheumatoid arthritis (p = 0.003), as well as high titers of anti-thyroid antibodies versus clinically healthy controls; 53% of thyroid ultrasounds were normal in hypothyroid patients, and increased perfusion was found in 40% of rheumatoid arthritis patients who tested positive for anti-thyroid antibodies. Cases classified as TIRADS 4 underwent aspiration with benign histopathological results. Conclusions: Thyroid assessment added clinical value was demonstrated in patients with rheumatoid arthritis, according to the prevalence of subclinical hypothyroidism, anti-thyroid antibodies positivity and ultrasound abnormalities, regardless of normal or altered thyroid function.

Characterization of B7H6, an endogenous ligand for the NK cell activating receptor NKp30, reveals the identity of two different soluble isoforms during normal human pregnancy.

B7H6, an endogenous ligand expressed on tumor cell surfaces, triggers NKp30-mediated activation of human NK cells. In contrast, the release of soluble B7H6 has been proposed as a novel mechanism by which tumors might evade NK cell-mediated recognition. Since NK cells are critical for the maintenance of early pregnancy, it is not illogical that soluble B7H6 might also be an important factor in directing NK cell activity during normal pregnancy. Thus, this study was focused on the characterization of soluble B7H6 during the development of normal pregnancy. Serum samples were obtained from healthy pregnant women who were experiencing their second pregnancies (n=36). Additionally, 17 of these pregnant participants were longitudinally studied for the presence of B7H6 during their second and third trimesters. Age-matched healthy non-pregnant women served as controls (n=30). The presence of soluble B7H6 was revealed by Western blotting. A further characterization was performed using an immunoproteomic approach based on 2DE-Western blotting combined with MALDI-MS. The results show that sera from all pregnant women were characterized by the presence of two novel isoforms of B7H6, both with lower MW than the reported of 51kDa. These isoforms were either a heavy (∼37kDa) or a light isoform (∼30kDa) and were mutually exclusive. N-glycosylation did not completely explain the different molecular weights exhibited by the two isoforms, as was demonstrated by enzymatic deglycosylation with PNGase F. The confirmation of the identity and molecular mass of each isoform indicates that B7H6, while maintaining the C- and N-termini, is most likely released during pregnancy by a mechanism distinct from proteolytic cleavage. We found that both isoforms, but mainly the heavier B7H6, were released via exosomes; and that the lighter isoform was also released in an exosome-free manner that was not observed in the heavy isoform samples. In conclusion, we find that soluble B7H6 is constitutively expressed during pregnancy and that, moreover, the soluble B7H6 is present in two new isoforms, which are released by exosomal and exosome-free mechanisms.

[Impact of pasteurization/freeze-drying on available immunoglobulin content of the mature human milk. Use in human milk banking of hospitals]. / Impacto de la pasteurización/liofilización en el contenido disponible de inmunoglobulinas en leche humana madura. Estudio de aplicación en bancos de leche humana en hospitales.

INTRODUCTION: This study analyzes the effect on the content of immunoglobulins and C3 complement of freeze drying after pasteurization by three different methods in mature human milk (MHM). OBJECTIVE: Freeze drying is proposed as a complementary method for the maintenance of MHM therapeutic properties with greater validity. METHODS: This was a descriptive study in which MHM samples were obtained. Next, aliquots of the samples obtained were pasteurized by three methods: 62.5 centigrades degrees/30 minutes, 72 centigrades degrees/15 minutes, 85 centigrades degrees/5 minutes, followed by a rapid cooling at 5 ºC. Then, 30 ml volumes of pasteurized sample were freeze-dried over a period of 36 hours. Total protein determination was performed by the Lowry method. The concentrations of immunoglobulins A, G and M, and complement C3, were determined by conventional nephelometric technique following the manufacturer's instructions. Statistical significance was defined as p < 0.05. RESULTS: The method of pasteurization of MHM with increased protein and immunoglobulin retention was at 62.5 centigrades degrees, however, pasteurization at 72 centigrades degrees before freeze-drying showed better retention of immunoglobulins. CONCLUSIONS: Our results suggest that the freeze-drying of pasteurized MHM is a suitable method for the conservation in human milk banks. Both the nutritional composition and the extension of its validity and the application of the two processes together provide the advantage of maintaining the therapeutic properties of human milk to improve the health of the newborn in a vulnerable, impaired or immunosuppressed state.

INTRODUCCIÓN: este estudio analiza el efecto sobre el contenido de inmunoglobulinas y complemento C3 de la liofilización posterior a la pasteurización por tres métodos diferentes en leche humana madura (LHM). OBJETIVO: la liofilización es propuesta como método complementario para el mantenimiento de las propiedades terapéuticas de la LHM con mayor vigencia. MÉTODOS: estudio descriptivo en el que se obtuvieron muestras de LHM. Alícuotas de las muestras obtenidas se pasteurizaron por tres métodos: 62,5 grados centígrados/30 minutos, 72 grados centígrados/15 minutos 85 grados centígrados/5 minutos, seguido de un enfriamiento rápido a 5 grados centígrados. Después, volúmenes de 30 ml de muestra pasteurizada fueron liofilizados durante un periodo de 36 horas. La determinación de proteínas totales fue realizada por el método Lowry. Las concentraciones de inmunoglobulinas A, G y M y el complemento C3 fueron determinadas por nefelometría convencional, siguiendo las instrucciones del fabricante. La significancia estadística se definió como p < 0,05. RESULTADOS: el método de pasteurización de LHM con mayor retención de proteína e inmunoglobulinas fue a la temperatura de 62,5 grados centígrados, sin embargo, la pasteurización a 72 grados centígrados antes de la liofilización mostró mayor retención de inmunoglobulinas. CONCLUSIONES: nuestros resultados sugieren que la liofilización de LHM pasteurizada es un método eficiente para la conservación en bancos de leche humana. Tanto la composición nutricional como la extensión de su vida útil y la aplicación de los dos procesos juntos proporcionan la ventaja de mantener las propiedades terapéuticas de la leche humana para mejorar la salud del recién nacido en estado vulnerable, desmedro o inmunosuprimido.

Disease duration of rheumatoid arthritis is a predictor of vascular stiffness: a cross-sectional study in patients without known cardiovascular comorbidities: A STROBE-compliant article.

The aim of this study was to analyze the impact of disease duration on carotid to femoral pulse wave velocity (cfPWV) in rheumatoid arthritis (RA) patients without either known traditional cardiovascular risk factors or previous comorbidities.Patients with RA diagnosis attending the rheumatology outpatient clinic of Hospital Civil Juan I. Menchaca, Guadalajara, Mexico, were analyzed. A total of 106 RA patients without known traditional cardiovascular risk factors were selected. All subjects were evaluated for RA disease duration, RA disease activity score on 28 joints (DAS28), serum lipids, rheumatoid factor and anti-cyclic citrullinated peptide (anti-CCP) antibodies. Arterial stiffness was measured as cfPWV by noninvasive tonometry. A multivariate regression model was used to analyze the contribution of RA disease duration and age on cfPWV. cfPWV was positively correlated with age (r = 0.450, P < .001), RA disease duration (r = 0.340, P < .001), total cholesterol (r = 0.312, P = .002), and low density lipoprotein (LDL-c) cholesterol (r = 0.268, P = .012). Patients with a RA disease duration ≥10 years exhibited significantly increased cfPWV compared with patients with disease duration <2 years (8.4 ±â€Š1.8 vs 7.0 ±â€Š0.8) and ≥2 to <10 years (8.4 ±â€Š1.8 vs 7.8 ±â€Š1.3), respectively. Age, RA disease duration, and triglycerides were predictors of cfPWV in multivariate analyses. According to the ß-coefficients, each year of disease duration (ß = 0.072) had a greater impact on cfPWV than age (ß = 0.054).Each year of life with RA contributes to a higher rate of vascular aging or stiffening than a year of life without RA. The cumulative damage provided by RA was most pronounced in patients with disease duration ≥10 years.

Impacto de la pasteurización/liofilización en el contenido disponible de inmunoglobulinas en leche humana madura. Estudio de aplicación en bancos de leche humana en hospitales / Impact of pasteurization/freeze-drying on available immunoglobulin content of the mature human milk. Use in human milk banking of hospitals

Introducción: este estudio analiza el efecto sobre el contenido de inmunoglobulinas y complemento C3 de la liofilización posterior a la pasteurización por tres métodos diferentes en leche humana madura (LHM). Objetivo: la liofilización es propuesta como método complementario para el mantenimiento de las propiedades terapéuticas de la LHM con mayor vigencia. Métodos: estudio descriptivo en el que se obtuvieron muestras de LHM. Alícuotas de las muestras obtenidas se pasteurizaron por tres métodos: 62,5 °C/30 minutos, 72 °C/15 minutos 85 °C/5 minutos, seguido de un enfriamiento rápido a 5 °C. Después, volúmenes de 30 ml de muestra pasteurizada fueron liofilizados durante un periodo de 36 horas. La determinación de proteínas totales fue realizada por el método Lowry. Las concentraciones de inmunoglobulinas A, G y M y el complemento C3 fueron determinadas por nefelometría convencional, siguiendo las instrucciones del fabricante. La significancia estadística se definió como p < 0,05. Resultados: el método de pasteurización de LHM con mayor retención de proteína e inmunoglobulinas fue a la temperatura de 62,5 °C, sin embargo, la pasteurización a 72 °C antes de la liofilización mostró mayor retención de inmunoglobulinas. Conclusiones: nuestros resultados sugieren que la liofilización de LHM pasteurizada es un método eficiente para la conservación en bancos de leche humana. Tanto la composición nutricional como la extensión de su vida útil y la aplicación de los dos procesos juntos proporcionan la ventaja de mantener las propiedades terapéuticas de la leche humana para mejorar la salud del recién nacido en estado vulnerable, desmedro o inmunosuprimido (AU)

Introduction: This study analyzes the effect on the content of immunoglobulins and C3 complement of freeze drying after pasteurization by three different methods in mature human milk (MHM). Objective: Freeze drying is proposed as a complementary method for the maintenance of MHM therapeutic properties with greater validity. Methods: This was a descriptive study in which MHM samples were obtained. Next, aliquots of the samples obtained were pasteurized by three methods: 62.5 °C/30 minutes, 72 °C/15 minutes, 85 °C/5 minutes, followed by a rapid cooling at 5 °C. Then, 30 ml volumes of pasteurized sample were freeze-dried over a period of 36 hours. Total protein determination was performed by the Lowry method. The concentrations of immunoglobulins A, G and M, and complement C3, were determined by conventional nephelometric technique following the manufacturer’s instructions. Statistical significance was defi ned as p < 0.05. Results: The method of pasteurization of MHM with increased protein and immunoglobulin retention was at 62.5 °C, however, pasteurization at 72 °C before freeze-drying showed better retention of immunoglobulins. Conclusions: Our results suggest that the freeze-drying of pasteurized MHM is a suitable method for the conservation in human milk banks. Both the nutritional composition and the extension of its validity and the application of the two processes together provide the advantage of maintaining the therapeutic properties of human milk to improve the health of the newborn in a vulnerable, impaired or immunosuppressed state (AU)

Detection of autoantibodies to DSF70/LEDGFp75 in Mexican Hispanics using multiple complementary assay platforms.

PURPOSE: Antinuclear autoantibodies (ANA) targeting the dense fine speckled antigen DFS70, also known as lens epithelium-derived growth factor p75 (LEDGF/p75), are attracting attention due to their low frequency in systemic rheumatic diseases but increased frequency in clinical laboratory referrals and healthy individuals (HI). These ANA specifically recognize the stress protein DFS70/LEDGFp75, implicated in cancer, HIV-AIDS, and inflammation. While their frequency has been investigated in various ethnic populations, there is little information on their frequency among Hispanics/Latinos. In this study, we determined the frequency of anti-DFS70/LEDGFp75 autoantibodies in Mexican Hispanics using multiple detection platforms. METHODS: The frequency of anti-DFS70/LEDGFp75 antibodies was determined in 171 individuals, including 71 dermatomyositis (DM) patients, 47 rheumatoid arthritis (RA) patients, 30 obesity (OB) patients, and 23 HI. Antibody detection was achieved using four complementary assay platforms: indirect immunofluorescence, Western blotting, ELISA, and chemiluminescent immunoassay. RESULTS: We detected relatively low frequencies of anti-DFS70/LEDGFp75 antibodies in patients with DM (1.4%), RA (4.3%), and OB (6.6%), and elevated frequency (17.4%) in HI. A strong concordance between the different antibody detection platforms was observed. CONCLUSIONS: The low frequency of anti-DFS70/LEDGFp75 antibodies in Mexican patients with rheumatic diseases, but relatively higher frequency in HI, is consistent with previous observations with non-Hispanic populations, suggesting that geographic differences or ethnicity do not influence the frequency of these autoantibodies. Our results also highlight the importance of confirmatory assays for the accurate detection of these autoantibodies. Future studies with larger cohorts of healthy Hispanics/Latinos are needed to confirm if their anti-DFS70/LEDGFp75 antibody frequencies are significantly higher than in non-Hispanics.

Disease Activity Score on 28 Joints and Polypharmacy Are Independent Predictors for Health-Related Quality of Life Evaluated by INCAVISA in Patients With Rheumatoid Arthritis.

OBJECTIVE: The aim of this study was to investigate the main factors associated to a diminished health-related quality of life (HRQoL) evaluated by INCAVISA (Health-Related Quality of Life Inventory for Latin American Patients) in patients with rheumatoid arthritis (RA). METHODS: Female, 18 years or older, RA (American College of Rheumatology 1987 criteria and American College of Rheumatology/European League against Rheumatism 2010 criteria) patients who attended the outpatient rheumatology department of the Hospital Civil "Dr. Juan I. Menchaca," Guadalajara, Mexico, matched with healthy controls were included. Patients with any known comorbidities or treatment with antidepressive drugs were excluded. Trained physicians performed the RA clinical evaluation and INCAVISA. All data were analyzed using SPSS 21.0 software (SPSS Inc, Chicago, IL); P < 0.05 was considered statistically significant. RESULTS: Patients with polypharmacy (≥3 drugs) had a lower HRQoL by INCAVISA. The number of drugs, total comorbidities, and DAS-28 (Disease Activity Score on 28 Joints) were negatively correlated with total INCAVISA. In multivariate analysis, DAS-28 and polypharmacy were independent predictors for a negative perception of HRQoL evaluated by INCAVISA in RA patients. CONCLUSIONS: Disease activity and disability secondary to RA have a negative impact in the HRQoL. Other factors such as the number of drugs prescribed to these patients have been shown to be important for the negative perception of their HRQoL evaluated by INCAVISA.

Low CD36 and LOX-1 Levels and CD36 Gene Subexpression Are Associated with Metabolic Dysregulation in Older Individuals with Abdominal Obesity.

Background. Obesity study in the context of scavenger receptors has been linked to atherosclerosis. CD36 and LOX-1 are important, since they have been associated with atherogenic and metabolic disease but not fat redistribution. The aim of our study was to determinate the association between CD36 and LOX-1 in presence of age and abdominal obesity. Methods. This is a cross-sectional study that included 151 healthy individuals, clinically and anthropometrically classified into two groups by age (<30 and ≥30 years old) and abdominal obesity (according to World Health Organization guidelines). We excluded individuals with any chronic and metabolic illness, use of medication, or smoking. Fasting blood samples were taken to perform determination of CD36 mRNA expression by real-time PCR, lipid profile and metabolic and low grade inflammation markers by routine methods, and soluble scavenger receptors (CD36 and LOX-1) by ELISA. Results. Individuals ≥30 years old with abdominal obesity presented high atherogenic index, lower soluble scavenger receptor levels, and subexpression of CD36 mRNA (54% less). On the other hand, individuals <30 years old with abdominal adiposity presented higher levels in the same parameters, except LOX-1 soluble levels. Conclusion. In this study, individuals over 30 years of age presented low soluble scavenger receptors levels pattern and CD36 gene subexpression, which suggest the chronic metabolic dysregulation in abdominal obesity.

Inverse Relationship of the CMKLR1 Relative Expression and Chemerin Serum Levels in Obesity with Dysmetabolic Phenotype and Insulin Resistance.

BACKGROUND: In obesity there is a subclinical chronic low-grade inflammatory response where insulin resistance (IR) may develop. Chemerin is secreted in white adipose tissue and promotes low-grade inflammatory process, where it expressed CMKLR1 receptor. The role of chemerin and CMKLR1 in inflammatory process secondary to obesity is not defined yet. METHODS: Cross-sectional study with 134 individuals classified as with and without obesity by body mass index (BMI) and IR. Body fat storage measurements and metabolic and inflammatory markers were measured by routine methods. Soluble chemerin and basal levels of insulin by ELISA and relative expression of CMKLR1 were evaluated with qPCR and 2(-ΔΔCT) method. RESULTS: Differences (P < 0.05) were observed between obesity and lean individuals in body fat storage measurements and metabolic-inflammatory markers. Both CMKLR1 expression and chemerin levels were increased in obesity without IR. Soluble chemerin levels correlate with adiposity and metabolic markers (r = 8.8% to 38.5%), P < 0.05. CONCLUSION: The increment of CMKLR1 expression was associated with insulin production. Increased serum levels of chemerin in obesity were observed, favoring a dysmetabolic response. The results observed in this study suggest that both chemerin and CMKLR1 have opposite expression in the context of low-grade inflammatory response manifested in the development of IR.

CCL2 Serum Levels and Adiposity Are Associated with the Polymorphic Phenotypes -2518A on CCL2 and 64ILE on CCR2 in a Mexican Population with Insulin Resistance.

Genetic susceptibility has been described in insulin resistance (IR). Chemokine (C-C motif) ligand-2 (CCL2) is overexpressed in white adipose tissue and is the ligand of C-C motif receptor-2 (CCR2). The CCL2 G-2518A polymorphism is known to regulate gene expression, whereas the physiological effects of the CCR2Val64Ile polymorphism are unknown. The aim of the study is to investigate the relationship between these polymorphisms with soluble CCL2 levels (sCCL2), metabolic markers, and adiposity. In a cross-sectional study we included 380 Mexican-Mestizo individuals, classified with IR according to Stern criteria. Polymorphism was identified using PCR-RFLP/sequence-specific primers. Anthropometrics and metabolic markers were measured by routine methods and adipokines and sCCL2 by ELISA. The CCL2 polymorphism was associated with IR (polymorphic A+ phenotype frequencies were 70.9%, 82.6%, in individuals with and without IR, resp.). Phenotype carriers CCL2 (A+) displayed lower body mass and fat indexes, insulin and HOMA-IR, and higher adiponectin levels. Individuals with IR presented higher sCCL2 compared to individuals without IR and was associated with CCR2 (Ile+) phenotype. The double-polymorphic phenotype carriers (A+/Ile+) exhibited higher sCCL2 than double-wild-type phenotype carriers (A-/Ile-). The present findings suggest that sCCL2 production possibly will be associated with the adiposity and polymorphic phenotypes of CCL2 and CCR2, in Mexican-Mestizos with IR.

Serum leptin and serum leptin/serum leptin receptor ratio imbalance in obese rheumatoid arthritis patients positive for anti-cyclic citrullinated peptide antibodies.

INTRODUCTION: Leptin has a prominent role in the development and maintenance of acute and chronic inflammatory states such as rheumatoid arthritis (RA) and obesity. Nevertheless, the association of serum leptin (sLep) and soluble leptin receptor (sLepR) in RA pathogenesis has not been clarified. The purpose of this study was to evaluate the association of sLep, sLepR and leptin production indexes such as sLep/fat mass ratio with clinical activity and biomarkers and anti-cyclic citrullinated peptide (anti-CCP) antibodies in RA compared with body mass index (BMI) matched control subjects. METHODS: We included 64 RA patients and 66 controls matched for age, gender and BMI. Subjects were evaluated for BMI, fat mass distribution, sLep, sLepR, sLep/fat mass ratio and sLepR/fat mass ratio. Patients were evaluated for clinical activity and anti-CCP antibodies. RESULTS: We found two or three fold increased sLep levels, sLep/sLepR ratio and sLep/fat mass ratio in obese anti-CCP positive RA patients vs. CONTROLS: Partial correlations showed that anti-CCP antibodies were correlated with sLep/fat mass ratio (partial r = 0.347, P = 0.033) after adjustment for age, subcutaneous adipose tissue and fat mass. CONCLUSIONS: In preobese and obese RA patients there is and increased production of sLep according to anti-CCP positivity. This phenomenon suggests there is an additive effect of chronic inflammation resulting from RA and obesity in which leptin favors the humoral response against citrullinated proteins. In summary, the data observed in our study suggests sLep could be a surrogate marker of chronicity and humoral immunity in RA in the presence of obesity.