Transcriptional regulation of brown adipose tissue hiperplasia and development

The colon and cervix cancer treatment is usually based on the activity of one or more chemotherapeutic agents, which stop the cell cycle in numerous ways, by showing a high level of adverse effects and the development of resistance. Therefore, finding new or improving the existing agents with potential antitumor activity is imperative in the contemporary pharmacotherapy of cancer. Increasing the efficiency of chemotherapeutic agents can be achieved by combining the treatment with chemopreventive agents, which would enhance the effectiveness of the cytostatics and reduce the side effects. The aim of this study is to investigate the effect of the alpha-lipoic acid, ketoprofen, meloxicam and the combinations of tese test substances with cisplatin and 5-fluorouracil, in the cell cultures of colon and cervical cancer and the primary cultures of the mononuclear cells of peripheral blood, on cell proliferation and the quantitative expression of NF-B, Bcl- 2 and Bax protein in order to assess the test substances’ chemopreventive potential. The MTT test in the cell proliferation assay was used to examine the effect of test substances on the proliferation of colon and cervix cancer cells, while the quantitative expression of the NF-B, Bcl-2 and Bax protein was determined in correspondence to the immunofluorescence method. The role of the mononuclear cells of peripheral blood in the conducted research, is based on monitoring the selectivity of the tested substances by analysing the expression of mitochondrial apoptosis markers. The research results show that the alpha-lipoic acid, ketoprofen and meloxicam have an important role in the chemoprevention of colon and cervix cancer, by having an influence on the expression of NF-B, Bcl-2 and Bax, as important markers of the apoptotic cell power of the colon and cervix cancer cells. The test substances showed a dose-dependent effect, especially in combination with the cytostatics. When it comes to the comparison of the antiiflamatory nonsteroidal drugs, the priority could be given to meloxicam, probably due to the more selective profile of the COX-2 inhibition. The results could be very good candidates for the further in vivo preclinical studies in order to potentiate the effects, in a particular reference to the development of resistance and the reduction of the side effects that occur in the standard cytostatic treatment of colon and cervix cance

Effects of methimazole-induced hypothyroidism on immunohistochemical, stereomorphometric and some ultrastructural characteristics of pancreatic β-cells

The function of pancreatic β-cells is to produce and secrete insulin, a crucial hormone in carbohydrate metabolism. The transcription factor PDX1 is required for insulin gene transcription and mature β-cell function. Since this factor is regulated by triiodothyronine, a disturbance in insulin biosynthesis and/or secretion is usually related to a deficiency of this hormone. In the present study, we used methods of immunohistochemistry, stereology and electron microscopy to explore the correlation between altered thyroid status and insulin synthesis/secretion in a model of methimazole-induced hypothyroidism in rats. In hypothyroid animals fewer functional PDX1-positive β-cells were detected in the islets of Langerhans, while insulin immunostaining was stronger. Stereological analysis of β-cell granules revealed more numerous immature insulin granules in hypothyroid rats. Taken together, these data suggest that the applied treatment caused impaired insulin synthesis and secretion. Rare cells with granules characteristic for both α- and β-cells observed in hypothyroid animals could provide functional compensation for diminished insulin synthesis

Calcium-SANDOZ®-induced erythrocyte exovesiculation and internalization of hemichromic material into rat brown adipocytes

An ultramicroscopic study of brown adipose tissue (BAT) of rats treated with Ca-SANDOZ® (480 mg/l) for 3 days, revealed erythrocyte exovesiculation and migratory erythrocytic complexes from the capillaries to adipocyte cytoplasm and mitochondria. Two types of erythrocytic material transfer were observed: (i) numerous exocytic vesicles with electron dense material leaving the erythrocytes; (ii) furcated complexes with microholes, embedded in amorphous material. The content of red blood cell (RBC) complexes passed through the capillaries and transferred to the brown adipocytes where it was detectable in the cytoplasm and mitochondria. Light microscopy confirmed sphenoechinocytic transformation of the RBCs in the blood smears of the Ca-SANDOZ® treated rats

Epigenetic Aspects of a New Probiotic Concept—A Pilot Study

Several studies report the important role of an altered gut microbiota in the development of obesity, highlighting the potential use of probiotics in the treatment of obesity. The aim of this study is to investigate the effect of a novel probiotic approach on the expression of specific miRNAs and mRNAs associated with obesity in combination with the hypocholesterolemic octacosanol. Twenty overweight/obese women participated in a randomized, placebo-controlled, double-blind study and were randomly divided into two groups: the intervention group (daily one capsule containing Lactobacillus plantarum 299v (DSM9843), Saccharomyces cerevisiae var. boulardii, and 40 mg octacosanol; N = 12) and the placebo group (N = 8). Changes in lipid parameters and expression of miRNAs and mRNAs were assessed before (T0) and after the 12-week intervention (T1). After the intervention, the expression of miR-155-5p (9.38 ± 0.85 vs. 8.38 ± 1.06, p = 0.05) and miR-24-3p (3.42 ± 0.38 vs. 2.71 ± 0.97, p = 0.031) showed significant decreases in the intervention group when compared to the control group. At T1, the expression of miR-155-5p (8.69 ± 1.31 vs. 9.3 ± 0.85, p = 0.04), miR-125b-5p (5.41 ± 1.18 vs. 5.99 ± 1.36, p = 0.049), and TNF-α (10.24 ± 1.66 vs. 11.36 ± 1.12, p = 0.009) were significantly decreased in the intervention group. No changes in lipids and anthropometric parameters were observed. The novel probiotic approach had a positive effect on regulating the expression of certain miRNAs and mRNAs important for regulating inflammation and adipogenesis, which are essential for obesity onset and control

Efekti suplementacije probiotikom na morfologiju krvnih pločica gojaznih žena i žena sa prokomernom težinom

Obesity could be associated with enhanced platelet (PL) function and thrombosis. Also, changes in gut microbiota composition were observed in obese patients, and probiotics supplementation has been shown to improve some of the obesity-associated metabolic parameters. This study aimed to determine the possible effects of probiotic supplementation on platelet morphology in overweight/obese premenopausal women.Twenty overweight (BMI = 25.0-29.9 kg/m2) and obese (BMI ≥ 30.0 kg/m2) women participated in double- blind, randomized, placebo-controlled study. They were randomly assigned to receive one capsule daily of probiotics (7x1010 CFU Lactobacillus plantarum 299v (DSM9843), 5x109 CFU Saccharomyces cerevisiae var. boulardii and 40mg octacosanol; N=10) or placebo (N=10) for 3 months. To determine changes in the parameters of platelet activation - the platelet count (PC), platelet distribution width (PDW), Feret diameter, and hypogranular appearance, blood samples were obtained for an automated hematology analyser and peripheral smears staining. Although difference in PC was not found between groups, supplementation with probiotics decreased PDW compared to the placebo group (p<0.05). Regarding platelets diameter, there is an decrease in group supplemented with probiotics compared to placebo, however, a statistical difference has not been reached. Further, the proportion of hypogranular PL was increased in a probiotic-supplemented group compared to the placebo group (p<0.01). In conclusion, probiotic supplementation achieved improvements in platelets morphology and hematological indices in overweight/obese premenopausal women.Gojaznost je često povezana sa povećanom funkcijom krvnih pločica i trombozom. Takođe, istraživanja ukazuju na promenjen sastav mikrobioma creva kod gojaznih osoba, kao i da suplementacija određenim probioticima može imati pozitivan uticaj na telesnu masu. Cilj rada je bio da se ispita uticaj suplementacije probiotskog preparata na morfološke karakteristike krvnih pločica u randomizovanoj, dvostruko-slepoj, placebo-kontrolisanoj studiji. U tu svrhu, regrutovano je 20 žena sa prekomernom težinom (BMI ≥ 25,0 kg/m2) koje su podeljene u dve grupe. Prva grupa je pila dnevno jednu kapsulu probiotskog preparata tokom tri meseca (kombinaciju soja Lactobacillus plantarum 299v (DSM9843), Saccharomyces cerevisiae var. boulardii i 40 mg oktakozanola; N = 10), dok je druga grupa bila placebo grupa. Određen je broj krvnih pločica, i njihova raspodela po volumenu (eng. platelet distribution width, PDW), a zatim je posmatrana morfologija krvnih pločica i izmeren njihov dijametar iz krvnih razmaza. Analiza je pokazala da su krvne pločice kod gojaznih/žena sa prekomernom težinom uglavnom krupne, što je jedan od pokazatelja njihove aktivnosti. U interventoj grupi nije zapažena promena u broju krvnih pločica, ali je značajno smanjen PDW u poređenju sa placebo grupom (p < 0,05). Dalje, pokazano je da je udeo hipogranularnih krvnih pločica bio povećan u poređenju sa placebo grupom (p < 0,01). Ovi rezultati ukazuju na pozitivan efekat probiotika na morfološke karakteristike krvnih pločica i mogućnost primene probiotika u regulaciji njihove aktivnosti u patološkim stanjima.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Expression and Subcellular Localization of Estrogen Receptors alpha and beta in Human Fetal Brown Adipose Tissue

Context: Brown adipose tissue (BAT) has the unique ability of generating heat due to the expression of mitochondrial uncoupling protein 1 (UCP1). A recent discovery regarding functional BAT in adult humans has increased interest in the molecular pathways of BAT development and functionality. An important role for estrogen in white adipose tissue was shown, but the possible role of estrogen in human fetal BAT (fBAT) is unclear. Objective: The objective of this study was to determine whether human fBAT expresses estrogen receptor alpha (ER alpha) and ER beta. In addition, we examined their localization as well as their correlation with crucial proteins involved in BAT differentiation, proliferation, mitochondriogenesis and thermogenesis including peroxisome proliferator-activated receptor gamma (PPAR gamma), proliferating cell nuclear antigen (PCNA), PPAR gamma-coactivator-1 alpha (PGC-1 alpha), and UCP1. Design: The fBAT was obtained from 4 human male fetuses aged 15, 17, 20, and 23 weeks gestation. ER alpha and ER beta expression was assessed using Western blotting, immunohistochemistry, and immunocytochemistry. Possible correlations with PPAR gamma, PCNA, PGC-1 alpha, and UCP1 were examined by double immunofluorescence. Results: Both ER alpha and ER beta were expressed in human fBAT, with ER alpha being dominant. Unlike ER beta, which was present only in mature brown adipocytes, we detected ER alpha in mature adipocytes, preadipocytes, mesenchymal and endothelial cells. In addition, double immunofluorescence supported the notion that differentiation in fBAT probably involves ER alpha. Immunocytochemical analysis revealed mitochondrial localization of both receptors. Conclusion: The expression of both ER alpha and ER beta in human fBAT suggests a role for estrogen in its development, primarily via ER alpha. In addition, our results indicate that fBAT mitochondria could be targeted by estrogens and pointed out the possible role of both ERs in mitochondriogenesis.Serbian Ministry of Education, Science, and Technological Development {[}173055, 173054