Firmicutes, Bacteroidetes and Actinobacteria in Human Milk and Maternal Adiposity.

The main objective was to explore the relationship between the microbiota of human milk and adiposity in Mexican mothers during the first lactation stage. METHODS: Seventy lactating women were included. Adiposity by anthropometric measurements and by bioelectric impedance was obtained. The donation of human milk was requested, from which bacterial DNA was extracted and qPCR of the 16S region was performed. The Mann-Whitney U test, Spearman and Pearson correlations, and multiple linear regressions models were also calculated. RESULTS: The median percentage of Bacteroidetes had a direct and significant correlation with normal adiposity, current BMI, waist circumference, and body fat percentage. The correlation with current BMI became significantly inverse in women with BMI ≥ 25. In women with normal BMI, the percentage of Actinobacteria showed a direct and significant correlation with current BMI, waist circumference, and percentage of body fat. Multiple linear regressions showed that pre-pregnancy BMI was the variable with the highest predictive value with the Bacteroidetes phyla in normal BMI and in BMI ≥ 25. CONCLUSIONS: the adiposity of the woman before pregnancy and during lactation would have an important effect on the abundance of Bacteroidetes and Actinobacteria in human milk.

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.

Using a lactation room at the workplace is associated with longer breastfeeding duration in working mothers.

INTRODUCTION: IIntroduction: maternal employment after childbirth is associated with abandonment of breastfeeding; however, lactation rooms in the workplace increase the practice of breastfeeding. Objective: to compare the frequency and duration of breastfeeding among working mothers based on the presence of lactation rooms in their workplaces. Methods: we included mothers from different institutions whose infants were between 6 and 35 months (n = 158), and an ad hoc questionnaire was applied to assess breastfeeding, exclusive breastfeeding (EBF), partial breastfeeding (PBF), and use of human milk substitutes (HMS). Two groups were compared: working mothers with a lactation room at their workplace (n = 76) versus working mothers without this resource (n = 82). Results: breastfeeding duration (7.5 vs. 5.0 months, p < 0.001) and EBF (3.0 vs. 1.2 months, p = 0.005) were higher in mothers who had a lactation room. HMS use was shorter in mothers who had a lactation room (2.5 vs. 10.0 months, p = 0.001). There were more working mothers who breastfed for more than six months (75.0 % vs. 48.8 %) [OR = 3.15 (95 % CI, 1.60-6.19), p = 0.001] and 12 months (31.6 % vs. 14.6 %) [OR = 2.69 (95 % CI, 1.23-5.87), p = 0.014] when lactation rooms were available in their workplaces. Conclusion: the presence of a lactation room in the workplace was associated with a higher frequency and duration of breastfeeding.

INTRODUCCIÓN: RESUMEN Introducción: el empleo materno después del parto se asocia con el abandono de la lactancia materna, mientras que las salas de lactancia en el lugar de trabajo aumentan la práctica de la lactancia. Objetivo: comparar la frecuencia y duración de la lactancia materna entre madres trabajadoras en función de la presencia o no de salas de lactancia en sus lugares de trabajo. Métodos: incluimos madres de diferentes instituciones cuyos bebés tenían entre 6 y 35 meses (n = 158) y se aplicó un cuestionario ad hoc para evaluar la lactancia materna, la lactancia materna exclusiva (LME), la lactancia materna parcial (LMP) y el uso de sucedáneos de la leche humana (SLH). Se compararon dos grupos: madres trabajadoras con una sala de lactancia en su lugar de trabajo (n = 76) y madres trabajadoras sin este recurso (n = 82). Resultados: la duración de la lactancia (7,5 vs. 5,0 meses, p < 0,001) y LME (3,0 vs. 1,2 meses, p = 0,005) fueron mayores en las madres que tenían sala de lactancia. El uso de SLH fue más corto en las madres que tenían sala de lactancia (2,5 vs. 10,0 meses, p = 0,001). Hubo más madres trabajadoras que amamantaron más de seis meses (75,0 % vs. 48,8 %) [OR = 3,15 (IC 95 %: 1,60-6,19), p = 0,001] y 12 meses (31,6 % vs. 14,6 %) [OR = 2,69 (IC 95 %: 1,23-5,87), p = 0,014] cuando había salas de lactancia disponibles en sus lugares de trabajo. Conclusión: la presencia de una sala de lactancia en el lugar de trabajo se asoció a una mayor frecuencia y duración de la lactancia materna.

[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.

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.

The 482Ser of PPARGC1A and 12Pro of PPARG2 Alleles Are Associated with Reduction of Metabolic Risk Factors Even Obesity in a Mexican-Mestizo Population.

The aim of this study was to investigate the relationship between functional polymorphisms Gly482Ser in PPARGC1A and Pro12Ala in PPARG2 with the presence of obesity and metabolic risk factors. We included 375 individuals characterized as Mexican-Mestizos and classified by the body mass index (BMI). Body dimensions and distribution of body fat were measured. The HOMA-IR and adiposity indexes were calculated. Adipokines and metabolic profile quantification were performed by ELISA and routine methods. Genetic polymorphisms were determined by polymerase chain reaction restriction fragment length polymorphism analysis. A difference between obese and nonobese subjects in polymorphism PPARGC1A distribution was observed. Among obese individuals, carriers of genotype 482Gly/Gly were observed to have decreased body fat, BMI, and body fat ratio versus 482Ser/Ser carriers and increased resistin and leptin levels in carriers Gly+ phenotype versus Gly- phenotype. Subjects with PPARG2 Ala- phenotype (genotype 12Pro/Pro) showed a decreased HOMA-IR index versus individuals with Ala+ phenotype (genotypes 12Pro/Ala plus 12Ala/Ala). We propose that, in obese Mexican-Mestizos, the combination of alleles 482Ser in PPARGC1A and 12Pro in PPARG2 represents a reduced metabolic risk profile, even when the adiposity indexes are increased.

The Impact of LEP G-2548A and LEPR Gln223Arg Polymorphisms on Adiposity, Leptin, and Leptin-Receptor Serum Levels in a Mexican Mestizo Population.

The polymorphisms in leptin (LEP G-2548A) and leptin-receptor (LEPR Gln223Arg) seem to influence obesity and lipid metabolism among others. The aim of this study was to investigate the effect of these polymorphisms on adiposity, leptin (sLeptin), and leptin-receptor (sLeptin-receptor) serum concentrations as well as inflammation markers. We included 382 adults originally from Western Mexico. They were genotyped by PCR-RFLP. Obese individuals showed higher sLeptin (58.2 ± 31.35 ng/mL) but lower sLeptin-receptor (12.6 ± 3.74 ng/mL) levels than normal weight ones (17.6 ± 14.62 ng/mL, 17.4 ± 4.62 ng/mL, resp.), P < 0.001. Obese subjects carriers of Arg/Arg genotype had more (P = 0.016) sLeptin-receptor (14.7 ± 4.96 ng/mL) and less (P = 0.004) sLeptin (44.0 ± 28.12 ng/mL) levels than Gln/Gln genotype (11.0 ± 2.92 ng/mL, 80.3 ± 33.24 ng/mL, resp.). Body fat mass was lower (P from 0.003 to 0.045) for A/A (36.5% ± 6.80) or Arg/Arg (36.8% ± 6.82) genotypes with respect to G/G (41.3% ± 5.52) and G/A (41.6% ± 5.61) or Gln/Gln (43.7% ± 4.74) and Gln/Arg (41.0% ± 5.52) genotypes carriers. Our results suggest that LEP -2548A and LEPR 223Arg could be genetic markers of less body fat mass accumulation in obese subjects from Western Mexico.

[Association of resistin gene 3'UTR+62G>A polymorphism with insulin resistance, adiposity and the adiponectin-resistin index in Mexican population]. / Asociación del polimorfismo 3'UTR+62G>A en resistina con resistencia a la insulina, adiposidad y el índice adiponectina-resistina en población mexicana.

INTRODUCTION: Insulin resistance (IR) is a disease with genetic susceptibility characterized by the increase in storage and irregular body fat distribution, and impaired production of adipokines. OBJECTIVE: The objective was to investigate the relationship between 3'UTR+62G>A RETN gene polymorphism, with adiponectin-resistin index (ARindex), adiposity, and inmuno-metabolic markers. METHODS: In this cross-sectional study, 260 individuals characterized as Mexican-Mestizo and classified in lean and overweight, and IR and without-IR, were included. Anthropometrics, body composition, body fat distribution and inflammation and metabolic markers were measured by routine methods, RETN 3'UTR+62G>A alleles were identified by PCR-RFLP and soluble insulin, total adiponectin and resistin were measured by ELISA methods. RESULTS: The +62G allele frequencies for lean and overweight individuals were different P=0.0343 (95.4% and 98.4%, respectively). The lean GA genotype carriers showed significant low measures of ARindex, adiposity, and inmuno-metabolic markers, than the GG genotype carriers. We found differences between individuals with IR and without-IR: in ARindex (P=0.002), adiponectin (P=0.002) and resistin levels (P=0.033): 1.102 ± 0.03, 5.167 ± 0.36 ug/mL and 8.827 ± 0.42 ng/mL versus 1.336 ± 0.07, 3.577 ± 0.34 ug/mL and 10.480 ± 0.65 ng/mL. Showed correlations with inflammation markers, distribution and body fat storage (r=0.262 to 0.414), P< 0.05. CONCLUSIONS: The present data suggest that in a Mexican-mestizo population the RETN +62G>A polymorphism is associated with overweight. The presence of the +62A allele was associated with increase of total adiponectin, ARindex, resistin levels, metabolic markers and body fat storage. ARindex can be an early indicator of insulin resistance.

Introducción: La resistencia a la insulina (RI) se caracteriza por susceptibilidad genética, incremento en la adiposidad y distribución irregular de grasa corporal, con alteración en la producción de adipocinas. Objetivo: Investigar la asociación del polimorfismo 3'UTR+62G>A en resistina con RI, índice adiponectina-resistina (ARindex), adiposidad y marcadores inmuno-metabólicos. Métodos: En un estudio transversal a 260 mestizos-mexicanos, clasificados con peso normal, exceso de peso, sin y con RI, se les evaluó: composición corporal, distribución de masa grasa y marcadores inmuno-metabólicos. Los alelos del polimorfismo 3'UTR+62G>A en resistina se identificaron por PCR-RFLP. La concentración sérica de insulina, adiponectina total y resistina se midieron por la técnica de ELISA. Resultados: Las frecuencias del alelo +62G para los individuos con peso normal y exceso de peso, fueron (95.4% y 98.4%, respectivamente) P=0.0343. Los portadores del genotipo GA con peso normal mostraron valores menores del ARindex, adiposidad y marcadores inmuno-metabólicos comparados con los portadores del genotipo GG. Se observó diferencia entre los individuos sin y con RI en el ARindex (P=0.002), concentración sérica de adiponectina (P=0.002) y resistina (P=0.033): 1.102±0.03, 5.167±0.36ug/mL y 8.827±0.42ng/mL versus 1.336±0.07, 3.577±0.34ug/mL y 10.480±0.65ng/mL, respectivamente. Los marcadores inmuno-metabólicos, reserva y distribución de grasa corporal correlacionan con ARindex (r=0.262 a 0.414), PA en los individuos mestizos-mexicanos con exceso de peso. El alelo +62A se asoció con incremento de adiponectina total, valores menores del ARindex, concentración de resistina, marcadores metabólicos y reserva de grasa corporal. El ARindex puede ser un indicador temprano de RI.

Association of ADIPOQ +45T>G polymorphism with body fat mass and blood levels of soluble adiponectin and inflammation markers in a Mexican-Mestizo population.

PURPOSE: Obesity is a disease with genetic susceptibility characterized by an increase in storage and irregular distribution of body fat. In obese patients, the decrease in the Adiponectin gene (ADIPOQ) expression has been associated with a systemic low-grade inflammatory state. Our aim was to investigate the relationship between ADIPOQ +45T>G gene simple nucleotide polymorphism (SNP rs2241766) with serum adiponectin (sAdiponectin), distribution of body fat storage, and inflammation markers. SUBJECTS AND METHODS: In this cross-sectional study, 242 individuals from Western Mexico characterized as Mexican-Mestizo and classified by body mass index (BMI), were included. Anthropometrics, body composition, body fat distribution, and inflammation markers were measured by routine methods. Genotypes were characterized using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique and sAdiponectin by the ELISA method. A P-value <0.05 was considered the statistically significant threshold. RESULTS: sAdiponectin is associated with BMI (P < 0.001) and the genotypes (P < 0.001 to 0.0046) GG (8169 ± 1162 ng/mL), TG (5189 ± 501 ng/mL), and TT (3741 ± 323 ng/mL), but the SNP ADIPOQ +45T>G is not associated with BMI. However, the detailed analysis showed association of this SNP with a pattern of fat distribution and correlations (P < 0.05) with inflammation markers and distribution of body fat storage (Pearson's r = -0.169 to -0.465) were found. CONCLUSION: In this study, we have suggested that the ADIPOQ +45G allele could be associated with distribution of body fat storage in obesity. On the other hand, as no association was observed between ADIPOQ +45T>G gene polymorphism and obesity, it cannot be concluded that the ADIPOQ +45G allele is responsible for the increase of adiponectin levels.