N-acetylcysteine inhibits lipid accumulation in mouse embryonic adipocytes.

Oxidative stress plays critical roles in the pathogenesis of diabetes, hypertension, and atherosclerosis; some authors reported that fat accumulation correlates to systemic oxidative stress in human and mice, but cellular redox environment effect on lipid accumulation is still unclear. In our laboratory we used mouse embryonic fibroblasts (undifferentiated cells: CC), which are capable of differentiating into mature adipocytes (differentiated cells: DC) and accumulate lipids, as obesity model. Here we analyzed the role of the well-known antioxidant and glutathione precursor N-acetylcysteine (NAC) in cellular MAPK modulation and lipid accumulation. We evaluated the effect of NAC on the adipogenic differentiation pathway using different doses: 0.01, 0.1, 1 and 5mM; no toxic doses in these cells. A dose of 5mM NAC [DCN-5] provoked a significant decrease in triglyceride accumulation (72±10 [DCN-5] vs 169±15 [DC], p<0.01), as well in Oil Red O stained neutral lipid content (120±2 [DCN-5] vs 139±12 [DC], p<0.01). Molecular mechanisms responsible for adipogenic differentiation involve increase of the expression of phosphoERK½ and phosphoJNK, 5mM NAC treatment inhibited both pERK½ and pJNK protein levels. We also evaluated the mitotic clonal expansion (MCE) which takes place during adipogenesis and observed an increase in DC at a rate of 1.5 cells number compared to CC at day 2, whereas the highest doses of NAC significantly inhibited MCE. Our results suggest that NAC inhibits lipid accumulation and the MAPK phosphorylation in mouse embryonic fibroblasts during adipogenic differentiation and further contribute to probe the importance of cellular redox environment in adipogenesis.

GAL3ST2 from mammary gland epithelial cells affects differentiation of 3T3-L1 preadipocytes.

INTRODUCTION: In the mammary gland, the involution that occurs when lactation ends is an important period for cancer development. We have previously demonstrated stromal-epithelium interactions evaluating conditioned medium of adipose tissue on breast epithelial metalloproteases activity (Creydt et al., Clin Transl Oncol 15:124-131, 2013). Here, we evaluated the effects of conditioned medium of breast epithelial mammary cells on stromal cells. MATERIALS AND METHODS: Conditioned medium from normal murine mammary gland cell line (NMuMG) and conditioned medium proteins were obtained. Then, they were evaluated on modulation of adipocyte differentiation, using 3T3-L1 cell line. RESULTS: We described, for the first time, that breast epithelial mammary cells could produce the enzyme galactose 3-O-sulfotransferase 2 (GAL3ST2). Importantly, GAL3ST2 is present in NMMuMG and two human breast cancer cell lines, and it is more strongly expressed in more metastatic tumors. When 3T3-L1 preadipocyte differentiation was triggered in the presence of conditioned medium from NMuMG or GAL3ST2, triglyceride accumulation was decreased by 40 % and C/EBPß expression by 80 % in adipocytes. In addition, the expression of FABP4 (aP2), another marker of adipocyte differentiation, was inhibited by 40 % in GAL3ST2-treated cells. CONCLUSIONS: Taken together, these results suggest that GAL3ST2 would interfere with normal differentiation of 3T3-L1 preadipocytes; raising the possibility that it may affect normal differentiation of stromal preadipocytes and be a link to tumor metastatic capacity.

N-Acetylcysteine affects obesity-related protein expression in 3T3-L1 adipocytes.

OBJECTIVES: Oxidative stress plays critical roles in the pathogeneses of diabetes, hypertension, and atherosclerosis, but its effect on fat accumulation is still unclear. In this study, we analyzed the role of the well-known antioxidant and a glutathione (GSH) precursor N-acetylcysteine (NAC) in fat accumulation and the expression of obesity-associated proteins. METHODS: We studied the effects of 10 µM NAC on obesity-related protein expression in cultured 3T3-L1 preadipocytes, which are able to differentiate into mature adipocytes and accumulate lipids. RESULTS: NAC treatment inhibited fat accumulation and reduced the expression of obesity-related proteins, including monoamine oxidase A, heat shock protein 70 (HSP70), aminoacylase -1 (ACY-1), and transketolase. DISCUSSION: Our results suggest that the effects of NAC on triglycerides (Tgs) and protein expression are correlated. In support of this, we showed that NAC treatment affected both the Tg synthesis pathway and the expression levels of proteins implicated in human obesity.

Perfil androgénico en mujeres hirsutas: Modificaciones de sus niveles en la fase lútea / Androgenic profile in hirsute women: Modifications of the levels in lutheal phase

Introducción: El objetivo de este estudio fue evaluar Los niveles de Testosterona (T), T libre TL, DHEAs y Androstanodiol glucuronidato (A2G) mujeres hirsutas con ciclos menstruales (CM) regulares en la fase folicular (FF) y en una muestra tomada entre -5 a -10 días premenstrual (FL) a los efectos de 1) poder definir bioquímicamente el tipo de hirsutismo y 2) determinar si el aumento de Progesterona modifica los niveles de los andrógenos. Materiales y Métodos: En 65 mujeres hirsutas con CM regulares se determinó en FF los niveles de T, A2G, y DHEAs por RIE y TL calculada por la ecuación de la ley de acción de masas, y en la FL los niveles de P4. En 28 de las 65 pacientes, en la FL se repitió el perfil androgénico Resultados: Los niveles de T correlacionaron, en todos los casos, con los de TL. En 51 de las pacientes los niveles de P4 fueron ovulatorios, 25 de las cuales tuvieron normales los andrógenos evaluados (Hirsutismo Idiopático) De las 26 pacientes restantes, en 2 tenían T aumentada, en 4 la DHEAS. Se obtuvieron 2 parámetros aumentados en los siguientes casos; en 2 la DHEAs y el A2G, en 1 la T y la DHEAs y en1 la T y el A2G. En 4 pacientes se obtuvieron incremento de los 3 parámetros. Estas pacientes corresponden a Hiperandrogénicas ovulatorias. Las 12 restantes de estas 26 hirsutas tenían solamente el A2G aumentados. Dado que éste constituye la expresión periférica de la 5alfa reductasa, las mismas podrían incluirse en el grupo de hiperandrogénicas ovulatorias por aumento local de DHT. En 14 de las 65 pacientes los niveles de P4 fueron compatibles con ciclos anovulatorios correspondiendo a pacientes con Síndrome de Ovario Poliquístico (SOP). En 6 de ellas se constató aumento de 1, 2 o los 3 parámetros evaluados (SOP hiperandrogénicos), en las restantes 6 pacientes los niveles androgénicos fueron normales (SOP con hirsutismo clínico). El A2G aumentó significativamente en FL en las mujeres con ciclos ovulatorios (4.89±2.19 vs 3.36±2.38 ng/ml en FL y FF, respectivamente). En las anovulatorias las diferencias no fueron significativas (4.32±3.16 vs 4.69±4.54 ng/ml en FL y FF, respectivamente. Estos resultados indican que la P4 podría inducir un incremento del A2G. Dado que la T no se modificó en la FL respecto a FF (0.28±0.22 vs 0.30±0.25ng/ml en hirsutas ovulatorias y 0.47±0.32 vs 0.42±0.23 en hirsutas anovulatorias) es posible que la P4 aumente el A2G por un camino distinto a la de la T y DHT Conclusiones: En base a estos resultados podemos concluir que la determinación de A2G podría ser empleada como parámetro complementario en el estudio del hiperandrogenismo debiendo realizarse en FF dado que en FL podría ser el resultado del metabolismo de hormonas no androgénicas.

Introduction: The aim of the present study was to evaluate the circulating levels of Testosterone (T), free T (TL), DHEAs and Androstanediol glucuronide (A2G) in hirsute women with regular menstrual cycles (CM) in follicular phase (FF), and in a samples obtained 5 to 10 days before the next menstrual bleeding (FL), in order to 1) biochemically define type of hirsutism and 2) determine whether the increase in progesterone (P4) induces changes in androgen levels. Materials and Methods: Sixty five hirsute women with regular CM were studied. FF levels of T, A2G and DHEAs were determined by RIA, and TL by mass law calculation. FL levels of P4 were measured by RIA. In 28 of the 65 patients the androgen profile was also evaluated in FL. Results: The levels of T correlated in every case with those of TL. In 51 patients P4 levels were ovulatory. Twenty five of them showed normal androgen levels (Idiopathic hirsutism). From the remaining 26 patients, 2 had increased T, and 4 had increased DHEAs. Two parameters were found increased in the following cases: DHEAs and A2G in 2, T and DHEAs in 1, and T and A2G in 1. All the 3 parameters were found increased in 4 cases. These patients were ovulatory hiperandrogenic women. The remaining 12 of these 26 hirsute women had only A2G increased. Since this steroid is the peripheral expression of the 5alpha reductase activity, these women could be included in the ovulatory hiperandrogenic group because of a local increase in DHT. In 14 of the 65 patients the levels of P4 correlated with anovulatory cycles corresponding to Polycystic Ovarian Syndrome (SOP). In 6 of them an increase of 1, 2 or the 3 parameters were observed (Hiperandrogenic SOP); in the remaining 6 patients androgen levels were normal (SOP with clinical hirsutism). FL A2G significantly increased in women with ovulatory cycles (4.89±2.19 vs 3.36±2.38 ng/ml in FL and FF, respectively. Differences were no significant in the anovulatory patients (4.32±3.16 vs 4.69±4.54 ng/ml in FL and FF, respectively. These results indicate that P4 could induce an increase in A2G. Since T did not change in FL respect to FF (0.28±0.22 vs 0.30±0.25ng/ml in ovulatory hirsute and 0.47±0.32 vs 0.42±0.23 in anovulatory hirsute) it is possible that P4 increases A2G through a pathway different than that of T and DHT. Conclusions: Based on these results we conclude that A2G could be used as a complementary parameter in the study of hiperandrogenism, only in FF since in FL, it could be the result of the metabolism of non-androgenic hormones.

Cell-cell communication between mouse mammary epithelial cells and 3T3-L1 preadipocytes: effect on triglyceride accumulation and cell proliferation

Interaction between parenchyma and stroma is essential for organogenesis, morphogenesis, and differentiation. Mammary gland has being the chosen model for developmental biologist because the most striking changes in morphology and function take place after birth. We have demonstrated a regulation of triglyceride accumulation by protein factors synthesized by normal mouse mammary gland epithelial cells (NMMG), acting on a cell line, 3T3-L1, long used as a model for adipogenesis. In this paper, we demonstrate that this inhibitory effect seems to be shared by other cells of epithelial origin but not by other cell types. We found a regulation of cell proliferation when NMMG cells are cultured in the presence of conditioned media from Swiss 3T3 or 3T3-L1 cells. We found a possible point of regulation for the mammary factor on a key enzyme of the lipid metabolic pathway, the glycerol-3-phosphate dehydrogenase. The inhibitory factor seems to have an effect on this enzymes activity and reduces it. The results presented herein contribute to the understanding of cell-cell communication in a model of a normal mammary gland.(AU)

Cell-cell communication between mouse mammary epithelial cells and 3T3-L1 preadipocytes: effect on triglyceride accumulation and cell proliferation

Interaction between parenchyma and stroma is essential for organogenesis, morphogenesis, and differentiation. Mammary gland has being the chosen model for developmental biologist because the most striking changes in morphology and function take place after birth. We have demonstrated a regulation of triglyceride accumulation by protein factors synthesized by normal mouse mammary gland epithelial cells (NMMG), acting on a cell line, 3T3-L1, long used as a model for adipogenesis. In this paper, we demonstrate that this inhibitory effect seems to be shared by other cells of epithelial origin but not by other cell types. We found a regulation of cell proliferation when NMMG cells are cultured in the presence of conditioned media from Swiss 3T3 or 3T3-L1 cells. We found a possible point of regulation for the mammary factor on a key enzyme of the lipid metabolic pathway, the glycerol-3-phosphate dehydrogenase. The inhibitory factor seems to have an effect on this enzyme's activity and reduces it. The results presented herein contribute to the understanding of cell-cell communication in a model of a normal mammary gland.

Telomerase activity in fine needle aspiration biopsy samples: application to diagnosis of human thyroid carcinoma.

BACKGROUND: The diagnosis of thyroid follicular carcinoma by fine needle aspiration biopsy is a well known problem in thyroid pathology. METHODS: We evaluated telomerase activity (TA) in 85 fine needle aspiration biopsy (FNAB) samples from patients with thyroid nodules. Surgery samples from patients with tumor or follicular adenomas were also analyzed. RESULTS: Twenty of the FNAB samples corresponded to carcinomas and were positive to telomerase assay (TA >10 Units). Among them, 4 follicular carcinomas and 1 papillary carcinoma were labeled as indeterminate by FNAB cytological examination. Four percent false positive cases and no false negative cases for TA in FNABs were reported. FNAB samples from follicular adenomas were diagnosed as indeterminate by cytological examination, but they showed no detectable TA. Tumor tissues from patients with follicular or papillary thyroid carcinomas presented TA >10 Units, whereas follicular adenoma tissues (benign nodules) showed no TA. CONCLUSION: Our results showed a good correlation between TA in FNAB samples and tumor/nodule thyroid tissue. This suggested that use of TA as a biological marker of malignancy might be a useful tool in the diagnosis of follicular thyroid carcinomas or follicular thyroid adenomas using FNAB samples.

Prevalencia de anemia y deficiencia de hierro en escolares jujeños de 12 años / Prevalence of anemia and iron deficiency in 12 year old school children from Jujuy

La deficiencia de hierro es una de las deficiencias de micronutrientes más comunes. Los adolescentes son un grupo vulnerable. Un reconocimiento oportuno puede prevenir una anemia ferropénica, etapa final y grave de dicha deficiencia, insuficientemente conocida en nuestro país. El objetivo de este estúdio fue determinar los valores hematológicos en adolescentes y conocer las prevalencias de anemia y deficiÛncia de hierro. Definidas como a) anemia: hematocrito (Hto) <38%, b) deficiencia componente funcional de hierro: saturación de transferrina <16%, c) deficiencia en los depósitos de hierro: ferritina (Ferr) <15 ng/ml. Se estudiaron 2.265 escolares de 12 años, de ambos sexos, en áreas urbanas y periurbanas de San Salvador de Jujuy (1.250 m.s.n.m.). Se determinaron: Hto, hierro sérico, capacidad total de ligadura de hierro a la transferrina por colorimetría y Ferr por ELISA. No se encontró anemia en la población estudiada. La ferrodeficiencia se observo en el componente funcional de hierro, en 25% de mujeres y 21% de varones y en depósitos de hierro em 28% de mujeres y 18% de varones. La deficiencia de los depósitos de hierro en ambos sexos es el dato de mayor interés. Los resultados confirman que la población estudiada constituye un grupo de riesgo. La detección temprana de la deficiencia de hierro contribuirá a un mejor desarrollo físico e intelectual de los adolescente. (AU)

Prevalencia de anemia y deficiencia de hierro en escolares jujeños de 12 años / Prevalence of anemia and iron deficiency in 12 year old school children from Jujuy

La deficiencia de hierro es una de las deficiencias de micronutrientes más comunes. Los adolescentes son un grupo vulnerable. Un reconocimiento oportuno puede prevenir una anemia ferropénica, etapa final y grave de dicha deficiencia, insuficientemente conocida en nuestro país. El objetivo de este estúdio fue determinar los valores hematológicos en adolescentes y conocer las prevalencias de anemia y deficiência de hierro. Definidas como a) anemia: hematocrito (Hto) <38%, b) deficiencia componente funcional de hierro: saturación de transferrina <16%, c) deficiencia en los depósitos de hierro: ferritina (Ferr) <15 ng/ml. Se estudiaron 2.265 escolares de 12 años, de ambos sexos, en áreas urbanas y periurbanas de San Salvador de Jujuy (1.250 m.s.n.m.). Se determinaron: Hto, hierro sérico, capacidad total de ligadura de hierro a la transferrina por colorimetría y Ferr por ELISA. No se encontró anemia en la población estudiada. La ferrodeficiencia se observo en el componente funcional de hierro, en 25% de mujeres y 21% de varones y en depósitos de hierro em 28% de mujeres y 18% de varones. La deficiencia de los depósitos de hierro en ambos sexos es el dato de mayor interés. Los resultados confirman que la población estudiada constituye un grupo de riesgo. La detección temprana de la deficiencia de hierro contribuirá a un mejor desarrollo físico e intelectual de los adolescente.

Bromocriptine and the expression of c-myc and c-fos in human prolactinomas.

Prolactinomas are one of the most frequent tumors of the human anterior pituitary. Dopamine agonists are the choice in the medical treatment of this disease. Bromocriptine (BC) is a well known anti-neoplasic agent in human PRL secreting adenomas although its effect on PRL cells is far from clear. We decided to investigate its influence on cell proliferation parameters: (3H)thymidine incorporation, expression of c-myc and c-fos, and number of estrogen receptors present in the samples. A total of 28 patients were included in this protocol. They were treated with BC (5-7.5mg day(-1) patient(-1)) or with vehicle orally 15 days before surgery. We found that in BC treated patients (3H)thymidine incorporation was lower than in vehicle treated patients. The oncogenes expression were diminished in BC comparing with vehicle-treated patients. No difference in the number of estrogen receptors was observed in the samples from BC or vehicle-treated patients. These results clearly demonstrate that one mechanism to reduce the size of human PRL secreting adenomas by BC is the inhibition of DNA duplication.

Antioxidants in the treatment of Graves disease.

An antioxidant mixture (LAROTABE) was evaluated in the treatment of Graves disease. Fifty-six hyperthyroid patients were treated with methimazol (MMI) (A), LAROTABE (B), or MMI plus LAROTABE (C). According to a clinical score, improvement was obtained at 8 weeks in A and 4 weeks in B and C. Group A diminished their thyroid hormone concentration to normal levels, while patients with LAROTABE did not reduce T3 and T4 unless MMI was introduced. Hyperthyroid patients had increased malondialdehyde (MDA) content and SOD activity and decreased catalase activity compared to controls. Within group A, MDA decreased to control values while SOD was reduced 38.3% and catalase increased 21.6%. Similar results were obtained for MDA and for both enzymes after treatment with LAROTABE. Signs and symptoms of Graves disease might be related to an increase in free radicals; antioxidants could be a new therapeutic tool to improve the clinical manifestation of this illness.