scholarly journals THE EFFECT OF DIET ENRICHED WITH PYROPHOSPHATE (E450) ON EXPRESSION OF GENES ENCODING BONE MORPHOGENETIC PROTEIN AND OSTEOCALCIN IN MOUSE EMBRYONIC MANDIBLE TISSUES

2021 ◽  
Vol 74 (1) ◽  
pp. 43-47
Author(s):  
Inessa I. Yakubova ◽  
Victor E. Dosenko ◽  
Lesya V. Tumanovska ◽  
Volodymyr I. Ostrianko
Keyword(s):  
Mrna Expression ◽  
Molecular Genetic ◽  
Mouse Embryos ◽  
Control Group ◽  
Molecular Genetic Study ◽  
Expression Of Genes ◽  
Morphogenetic Protein ◽  
Genes Encoding ◽  
Materials Used ◽  
The Impact

The aim: Of our study was to measure the mRNA expression of the investigated odontogenesis factors in mandible tissue of mouse embryos (17th day of pregnancy) gestated by females, kept on a E450 rich diet since 30 days before fertilization to gestation. Materials and methods: The effect of food supplements was studied in «Overload phosphates model». Experiments were carried out on white nonlinear outbred mice with mass 25−28g (n=40). The females from the control group were fed with standard rodent food, whereas the experimental females were fed with pyrophosphate-enriched food. The materials, used for the molecular genetic study, were the lower jaws of 17-days old mouse embryos (E−17). Results: The investigated BMP2 and osteocalcin genes are expressed at approximately the same level. Pyrophosphate-rich diet does not alter BMP2 gene expression, but it significantly increases the expression of osteocalcin. Conclusions: The present study is the first one to describe the impact of the pyrophosphate-rich diet on mRNA expression of key osteogenesis regulators – osteocalcin and BMP2.

scholarly journals Association of the I/D polymorphism of angiotensinconverting enzyme gene with the development of essential hypertension

2019 ◽  
Vol 34 (3) ◽  
pp. 87-96
Author(s):  
O. S. Pavlova ◽  
S. E. Ogurtsova ◽  
M. M. Liventseva ◽  
T. H. Lakotko ◽  
I. Y. Korobko ◽  
...  
Keyword(s):  
Essential Hypertension ◽  
Molecular Genetic ◽  
Recessive Model ◽  
Control Group ◽  
Molecular Genetic Study ◽  
Ace Gene ◽  
Paternal Line ◽  
Male Patients ◽  
Polymerase Chain ◽  
The Impact

Objective. To determine the impact of the I/D polymorphism of the angiotensin-converting enzyme (ACE) gene on the development of essential hypertension, taking into account gender differences.Material and Methods. Clinical data were assessed and a molecular genetic study was performed in 602 people including 401 patients with essential hypertension and 201 individuals of the control group, representing the Belarusian ethnic group. Genotyping was performed using the method of polymerase chain reaction and restriction fragment length polymorphism.Results. The distribution of genotypes of the I/D polymorphism of the ACE gene did not differ between patients with hypertension and normotensive individuals: II, ID, and DD genotypes were detected in 100 (24.9%), 192 (47.9%), and 109 (27.2%) patients and in 52 (25.9%), 108 (53.7%), and 41 (20.4%) people of the comparison group, respectively. Differences were found between the distribution of DD genotype in men with hypertension and in the control group, where the frequencies were 28.4% and 17.3% (p  =  0.04), respectively, in contrast to no differences in women: 25.8% and 23.3% (p  =  0.64), respectively. Carrying the DD genotype in men compared with the ID and DD genotypes (recessive model) of the I/D polymorphism of the ACE gene increased the probability of developing essential hypertension by 1.9 times (OR   =   1.89; 95% CI  =  1.04-3.44). The analysis of the prevalence of risk factors depending on the I/D polymorphism of the ACE gene showed that male patients with the DD genotype more often had burdened heredity in regard to the development of premature cardiovascular diseases (23 patients (37.7%)) compared with the individuals with II and ID genotypes: 13 (21.7%) and 14 (14.9%) patients, respectively (χ2  =  1.16; p  =   0.005), and mainly through the paternal line.Conclusions. Development of essential hypertension is associated with the carriership of the mutant DD genotype of I/D polymorphism of the ACE gene in men. 

Individual and Combined Assessment of Ser680Asn FSH Receptor and FSHβ -211 G>T Gene Polymorphisms in Ovarian Response in IVF/ICSI Program

2020 ◽  
Vol 21 ◽  
Author(s):  
Elli Anagnostou ◽  
Alexia Kafkoutsou ◽  
Despina Mavrogianni ◽  
Ekaterini Domali ◽  
Evangelia Dimitroulia ◽  
...  
Keyword(s):  
Gene Polymorphism ◽  
Gene Polymorphisms ◽  
Ovarian Response ◽  
Greek Population ◽  
Control Group ◽  
Genotype Distribution ◽  
Β Subunit ◽  
Molecular Genetic Study ◽  
The Impact ◽  
First Time

Background: Molecular biology tools, such as the detection of single nucleotide polymorphisms (SNPs), have been considered to assist to the management of the ovarian stimulation protocols. Purpose: The aim of this study was to evaluate the impact of two polymorphisms, the Asn680Ser polymorphism of the FSHR gene, and the FSH β subunit (FSHβ) gene polymorphism -211 G>T, in a Greek population of women undergoing IVF/ICSI program in our center. In addition, a control group of fertile women was studied, to verify whether there are differences in the genotype distribution between fertile and infertile population for both polymorphisms, as the FSHβ gene polymorphism -211 G>T is studied for the first time in the Greek population. Results : The FSH β-211 G>T polymorphism, studied for the first time in the Greek infertile population, appears to be quite rare. When studying the two polymorphisms separately, statistically significant differences were obtained that concerned the LH levels. Discussion: According to the combination analysis of the two polymorphisms by the number of alleles, women with 2-3 polymorphic alleles needed more days of stimulation, but there were no differences in pregnancy rates. Conclusion: This molecular genetic study helps to elucidate whether the polygenic combination of the Asn680Ser and FSH β subunit -211 G>T gene polymorphisms is of additive value in the prediction of ovarian response to exogenous gonadotropins.

scholarly journals The Assessment of IL-21 and IL-22 at the mRNA Level in Tumor Tissue and Protein Concentration in Serum and Peritoneal Fluid in Patients with Ovarian Cancer

2021 ◽  
Vol 10 (14) ◽  
pp. 3058
Author(s):  
Aleksandra Mielczarek-Palacz ◽  
Celina Kruszniewska-Rajs ◽  
Marta Smycz-Kubańska ◽  
Jarosław Strzelczyk ◽  
Wojciech Szanecki ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Peritoneal Fluid ◽  
Tumor Tissue ◽  
Mrna Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
First Time ◽  
Ovarian Serous Cancer

The aim of the analysis was for the first time to assess the expression of genes encoding IL-21 and IL-22 at the mRNA level in ovarian tumor specimens and the concentration of these parameters in serum and peritoneal fluid in patients with ovarian serous cancer. The levels of IL-21 and IL-22 transcripts were evaluated with the use of the real-time RT-qPCR. Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration of proteins. Quantitative analysis of IL-21 gene mRNA in the tumor tissue showed the highest activity in the G1 degree of histopathological differentiation and was higher in G1 compared to the control group. The concentration of IL-21 and IL-22 in the serum and in the peritoneal fluid of women with ovarian cancer varied depending on the degree of histopathological differentiation of the cancer and showed statistical variability compared to controls. The conducted studies have shown that the local and systemic changes in the immune system involving IL-21 and IL-22 indicate the participation of these parameters in the pathogenesis of ovarian cancer, and modulation in the IL-21/IL-22 system may prove useful in the development of new diagnostic and therapeutic strategies used in patients, which require further research.

scholarly journals MOLECULAR GENETIC STUDY OF THE IMPACT OF SNP C677T OF THE GENE MTHFR IN THE DEVELOPMENT OF CONGENITAL ISOLATED CLEFT LIP AND PALATE

2018 ◽  
Vol 25 (5) ◽  
pp. 104-110
Author(s):  
V. S. Uchaeva ◽  
Yu. A. Vasiliev ◽  
A. S. Gracheva ◽  
O. V. Gulenko ◽  
I. G. Udina
Keyword(s):  
Genetic Study ◽  
Cleft Lip ◽  
Cleft Lip And Palate ◽  
Control Group ◽  
Population Genetic Study ◽  
Molecular Genetic Study ◽  
Increased Risk ◽  
Significant Difference ◽  
Congenital Cleft ◽  
The Impact

Aim. This research was designed to conduct an associative population genetic study for the consideration of the impact of SNP C677T of the gene MTHFR in the congenital maxillofacial developmental anomalies (CMDA): congenital cleft lip (CCL), congenital cleft palate (CCP), congenital cleft lip and palate (CCLP) in the Krasnodar territory. The aim of the study is to establish the associations between SNP C677T of the gene MTHFR and the development of congenital cleft lip and/ or palate.Materials and methods. In this research, the peculiarities of distribution of SNP C667T of the gene MTHFR in children with congenital cleft lip and/or palate (n=223) and their mothers (n=78) in comparison with the control group (n=124) were studied in the Krasnodar territory. The genetic demographic questionnaires were gathered for children with CMDA, the information about diagnosis was obtained from the medical records. The biological samples, including blood or scrapings of oral mucosa, were collected from children with the pathology and their mothers. The DNA was extracted from the samples by the standard method. The study of the peculiarities of distribution of alleles of SNP C677T of the gene MTHFR was performed by PCR-PFLP with endonuclease Hinf I or by tetra-primer ARMS-PCR method in children with CCL, CCP, CCLP, their mothers and the control group. Statistical processing of the obtained data was performed by the algorithms of the “Statistica” program.Results. While comparing the profiles of frequencies of SNP C677T in children with CCL, CCP and CCLP with the control group, there were identified no significant differences in the frequency of this SNP and no peculiarities of genotypes distribution. There was identified a significant difference in the peculiarities of genotypes distribution with the control group (G=19,5232, d.f.=1, p<0,001) as well as united genotypes (С/C и С/T) in accordance to T/T (G=10,4657, d.f.=1; p<0,001) and united genotypes (C/T и T/T) in accordance to C/C (G=15,1896, d.f.=1, p<0,001) for the mothers of children with CCL, CCP and CCLP.Conclusion. As a result of the study, we established the association of SNP C677T of the MTHFR gene with the development of congenital cleft lip and/or palate: mothers’ T/T genotype is associated with the increased risk of giving birth to a child with CCL, CCP and CCLP (in comparison with mothers with C/C+C/T genotype): odds ratio [OR]=16,63, 95% CI: 3,86-71,71; p=0,0003 and also for mothers with genotypes (C/T+T/T) in comparison with mothers with genotypes C/C: OR=3,22, CI:1,71-6,08; p=0,0002. The amount of risk is not significant in children with CMDA for T/T genotype. So it is possible to make a conclusion about the impact of C677T of the gene MTHFR in the development of CCL, CCP and CCLP only in mother’s genotype. 

scholarly journals GENETIC ASPECTS OF PREGNANCY MISCARRIAGE

2019 ◽  
pp. 71-76
Author(s):  
K. M. Lisova ◽  
I. V. Kalinovska ◽  
O. M. Yuzko
Keyword(s):  
Pregnant Women ◽  
Molecular Genetic ◽  
Clinical Signs ◽  
Clinical Manifestations ◽  
Control Group ◽  
Molecular Genetic Study ◽  
Immunological Parameters ◽  
Fetoplacental Complex ◽  
Maternal Genotype ◽  
A1166c Polymorphism

Pregnancy miscarriage is a consequence of many factors. The aim of the study was to analyze the effect of miscarriage gene on embryometric, ultrasound, hormonal, immunological parameters in pregnant women, and to evaluate its prognostic value. The main group includes 31 pregnant women who had clinical signs of miscarriage in current or previous pregnancy. The control group consists of 32 healthy pregnant women whose clinical-paraclinical parameters served as a control to compare the data of the pregnancy survey of the main surveillance group. A general clinical examination and a special obstetrical examination (complaints, anamnesis, general medical examination, obstetric examination), biochemical studies (determination of hormones of the fetoplacental complex in blood serum of pregnant women), ultrasound, immunological studies, histological studies of the placenta, molecular genetic study A1166C polymorphism of the AGTR1 gene were made. In the course of the research, the genetic determinism of miscarriage was discovered. The polymorphism of the A1166C of the AGTR1 gene was considered as a prognostic marker of miscarriage in early gestational term and preeclampsia in the second half of pregnancy. A reliable marker of abortion was the maternal genotype 1166AC for the genome AGTR1. The risk of occurrence of clinical manifestations of abortion increased five times. At simultaneous influence of all prognostic factors the risk of abortion increased 6,25 times. Detection of genetic markers of pregnancy miscarriage will allow early correction of this pathology and prevent perinatal loss.

scholarly journals Increased Growth Differentiation Factor 15 in Patients with Hypoleptinemia-Associated Lipodystrophy

2020 ◽  
Vol 21 (19) ◽  
pp. 7214 ◽  
Author(s):  
Susan Kralisch ◽  
Annett Hoffmann ◽  
Juliane Estrada-Kunz ◽  
Michael Stumvoll ◽  
Mathias Fasshauer ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Transforming Growth Factor ◽  
Linear Regression Analysis ◽  
Control Group ◽  
Serum Concentrations ◽  
Metabolic Complications ◽  
Growth Differentiation Factor 15 ◽  
Differentiation Factor ◽  
Obesity And Diabetes ◽  
The Impact

Objective. Similar to obesity, lipodystrophy (LD) causes adipose tissue dysfunction and severe metabolic complications. Growth differentiation factor 15 (GDF15) belongs to the transforming growth factor β superfamily and is dysregulated in metabolic disease including obesity and diabetes mellitus. Circulating levels in LD and the impact of leptin treatment have not been investigated so far. Material and Methods. GDF15 serum levels were quantified in 60 LD patients without human immunodeficiency virus infection and 60 controls matched for age, gender, and body mass index. The impact of metreleptin treatment on circulating GDF15 was assessed in a subgroup of patients. GDF15 mRNA expression was determined in metabolic tissues of leptin-deficient lipodystrophic aP2-nSREBP1c-Tg mice, obese ob/ob mice, and control C57Bl6 mice. Results. Median GDF15 serum concentrations were significantly higher in LD patients (819 ng/L) as compared to the control group (415 ng/L) (p < 0.001). In multiple linear regression analysis, an independent and positive association remained between GDF15 on one hand and age, patient group, hemoglobin A1c, triglycerides, and C-reactive protein on the other hand. Moreover, there was an independent negative association between GFD15 and estimated glomerular filtration rate. Circulating GDF15 was not significantly affected by metreleptin treatment in LD patients. Gdf15 was upregulated in leptin-deficient lipodystrophic mice as compared to controls. Moreover, Gdf15 mRNA expression was downregulated by leptin treatment in lipodystrophic and obese animals. Conclusions. Serum concentrations of GDF15 are elevated in LD patients and independently associated with markers of metabolic dysfunction. Gdf15 expression is higher in lipodystrophic mice and downregulated by leptin treatment.

153 SUPPLEMENTATION WITH LINOLENIC ACID AND L-CARNITINE DURING IVM REDUCED THE EXPRESSION OF GENES RELATED TO LIPOGENESIS BUT DID NOT ALTER THE LIPID CONTENT AND CRYOTOLERANCE OF IN VITRO-PRODUCED EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 185 ◽  
Author(s):  
B. C. S. Leao ◽  
N. A. S. Rocha Frigoni ◽  
P. C. Dall'Acqua ◽  
M. Ambrogi ◽  
G. B. Nunes ◽  
...  
Keyword(s):  
Gene Expression ◽  
Linolenic Acid ◽  
Lipid Content ◽  
Control Group ◽  
Intracellular Lipid ◽  
Chi Square ◽  
Sudan Black B ◽  
Expression Of Genes ◽  
The Impact

This study was conducted to evaluate the impact of supplementation during in vitro maturation (IVM) with linolenic acid (ALA), l-carnitine (L-car), or the combination of both supplements on the embryo intracellular lipid content and cryotolerance, as well as in the embryo expression of genes involved in lipid metabolism (lipogenesis regulation: SCD1, FASN, and SREBP1; and β-oxidation pathway: CPT1B and CPT2). Cumulus-oocyte complexes (n = 1076) were IVM for 22 h at 38.5°C and 5% CO2 in air, in TCM-199 medium with bicarbonate, hormones, and 10% FCS (control group), supplemented with 100 μM ALA (ALA group), 5 mM L-car (L-car group), or a combination of 100 μM ALA + 5 mM L-car (ALA + L-car group). After IVF, presumptive zygotes were in vitro cultured in SOFaa medium supplemented with 5 mg mL−1 BSA and 2.5% FCS, at 38.5°C and 5% CO2 in air during 7 days. Cleavage and blastocyst rates were evaluated on Day 3 and 7, respectively (IVF = Day 0). At Day 7, the blastocysts were stained with the lipophilic dye Sudan Black B (n = 60), vitrified/warmed (n = 260; Ingámed® protocol, Maringa-PR, Brazil), or collected for analysis of gene expression (n = 180). Embryonic development were analysed by ANOVA and the multiple comparisons of means were determined by Tukey’s test. The embryonic re-expansion data were subjected to chi-square test and the differences in gene expression among groups were evaluated by Duncan’s multiple range test (P < 0.05). Data are presented as means ± standard error means. There was no effect (P > 0.05) of the supplements used during IVM on cleavage (79.54 ± 2.76% to 82.16 ± 1.13%) and blastocyst rates (29.03 ± 3.07% to 30.46 ± 2.01%). Similarly, the intracellular lipid content in Day-7 blastocysts (1.03 ± 0.04 to 1.15 ± 0.07 pixels) and the embryonic cryotolerance, assessed by the re-expansion rates after 24 h (67.3 to 78.3%) hatching rates after 48 h (11.5 to 25.5%) of post-warming culture, were unaffected (P > 0.05) by the supplements of IVM medium. Although the treatments did not alter (P > 0.05) the expression of CPT1B and CPT2 genes, the expression of FASN gene was decreased (P < 0.05) in the ALA group and the expression of SREBP1 gene was decreased (P < 0.05) in the ALA and L-car groups. The expression of the gene SCD1 was reduced (P < 0.05) in all treatments compared with the control group. Thus, despite the lack of effects of the treatments performed during IVM on the intracellular lipid content and cryotolerance of the embryos derived from the treated oocytes, a reduction in the expression of genes related to lipogenesis was observed in Day-7 blastocysts. These results suggest that treatments performed in the oocytes during IVM may have prolonged effects, affecting the subsequent expression of genes in embryos. Further studies are needed to determine the mechanisms related to the differentiation of the oocyte machinery during maturation. Financial support was provided by FAPESP (#2012/10084–4 and #2013/07382–6).

scholarly journals Role of hyaluronan in regulating self-renewal and osteogenic differentiation of mesenchymal stromal cells and pre-osteoblasts

2020 ◽  
Vol 24 (11) ◽  
pp. 3923-3937
Author(s):  
Maria B. Asparuhova ◽  
Vivianne Chappuis ◽  
Alexandra Stähli ◽  
Daniel Buser ◽  
Anton Sculean
Keyword(s):  
Growth Factor ◽  
Osteogenic Differentiation ◽  
Bone Matrix ◽  
Osteogenic Potential ◽  
Differentiation Markers ◽  
Self Renewal ◽  
Downstream Signaling ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
The Impact

Abstract Objectives The aim of the study was to investigate the impact of two hyaluronan (HA) formulations on the osteogenic potential of osteoblast precursors. Materials and methods Proliferation rates of HA-treated mesenchymal stromal ST2 and pre-osteoblastic MC3T3-E1 cells were determined by 5-bromo-20-deoxyuridine (BrdU) assay. Expression of genes encoding osteogenic differentiation markers, critical growth, and stemness factors as well as activation of downstream signaling pathways in the HA-treated cells were analyzed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunoblot techniques. Results The investigated HAs strongly stimulated the growth of the osteoprogenitor lines and enhanced the expression of genes encoding bone matrix proteins. However, expression of late osteogenic differentiation markers was significantly inhibited, accompanied by decreased bone morphogenetic protein (BMP) signaling. The expression of genes encoding transforming growth factor-β1 (TGF-β1) and fibroblast growth factor-1 (FGF-1) as well as the phosphorylation of the downstream signaling molecules Smad2 and Erk1/2 were enhanced upon HA treatment. We observed significant upregulation of the transcription factor Sox2 and its direct transcription targets and critical stemness genes, Yap1 and Bmi1, in HA-treated cells. Moreover, prominent targets of the canonical Wnt signaling pathway showed reduced expression, whereas inhibitors of the pathway were considerably upregulated. We detected decrease of active β-catenin levels in HA-treated cells due to β-catenin being phosphorylated and, thus, targeted for degradation. Conclusions HA strongly induces the growth of osteoprogenitors and maintains their stemness, thus potentially regulating the balance between self-renewal and differentiation during bone regeneration following reconstructive oral surgeries. Clinical relevance Addition of HA to deficient bone or bony defects during implant or reconstructive periodontal surgeries may be a viable approach for expanding adult stem cells without losing their replicative and differentiation capabilities.

Proteasome Inhibitors and a Fusicoccin Derivative (ISIR-042) Cooperatively Inhibit Proliferation of Myeloma Cells through Induction of C/EBP-Homologous Protein (CHOP/GADD153)

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 5657-5657
Author(s):  
Takaaki Miyake ◽  
Yoshio Honma ◽  
Tsutomu Takahashi ◽  
Koshi Kawakami ◽  
Takahiro Okada ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mrna Expression ◽  
Research Funding ◽  
Synergistic Effects ◽  
Proteasome Inhibitors ◽  
Cytotoxic Effects ◽  
Myeloma Cells ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Human Multiple Myeloma

Abstract Combinations of bortezomib and novel targeted therapeutic agents may synergistically increase antitumor effect and may overcome specific cellular resistance and/or antiapoptotic processes. However, highly effective combinations of bortezomib with targeted therapeutic agents have not been developed to date. We examined the synergistic effects of various compounds and bortezomib on myeloma cells and identified fusicoccin derivative ISIR-042 as the most potent drug. We developed ISIR-042 that exerts higher cytotoxic effects on hypoxic cells than on normoxic cells and that preferentially inhibits stem/progenitor cells in pancreatic cancer cell lines compared with other chemotherapeutic agents (Kawakami et al, Anticancer Agents Med Chem 2012). In the present study, we determined the synergistic effects of bortezomib and ISIR-042 cotreatment on human multiple myeloma cells. Human multiple myeloma cell lines RPMI8226, KMS-11, and KMS-26 were cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere of 5% CO2 in air. Viability of cells cultured with or without the test compounds for indicated time was examined by performing a modified MTT assay.Bone marrow specimens and ascitic fluids were collected from patients at diagnosis or relapse after obtaining written informed consent for sample collection in accordance with institutional policy. The study was approved by the Institutional Review Board of Shimane University. For performing colony-forming assay, the cells (1 × 104/dish) were plated in 1.1 ml semisolid methylcellulose medium supplemented with 0.8% methylcellulose and 20% FBS in triplicates for 10 days. CHOP-/- and CHOP+/+ MEF cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% FBS. Expression of cell surface antigens was analyzed by performing flow cytometry with anti-CD38 or anti-CD138 antibody. Combined effects of myeloma cells in primary culture were analyzed by performing flow cytometry with anti-CD19 and anti-CD138 antibodies. Expression of caspase-10, BCL-2, p62, BCLAF1, LC3, actin, and CHOP was determined by performing western blotting with respective monoclonal antibodies. mRNA expression of genes encoding XBP-1u and XBP-1s was analyzed by performing reverse transcription-PCR. CHOP mRNA levels were measured by performing real-time PCR. ISIR-042 inhibited the growth of RPMI8226 cells in a concentration-dependent manner and exerted synergistic effects with bortezomib. ISIR-042 also exerted synergistic effects with carfilzomib. Results of flow cytometry showed that the combination of ISIR-042 and bortezomib decreased the number of myeloma cells in the primary culture in a concentration-dependently. Colony-forming assay showed that RPMI8226 cells were more sensitive than normal mouse hematopoietic cells to ISIR-042 and bortezomib cotreatment. ISIR-042 decreased the number of CD38-CD138- cells and increased the number of CD38+CD138+ cells in KMS11 cell population. mRNA expression of genes encoding XBP-1u and XBP-1s was not affected in RPMI8226 cells treated with ISIR-042 or bortezomib alone. However, mRNA expression of the gene encoding XBP-1s significantly increased in RPMI8226 cells cotreated with ISIR-042 and bortezomib. Growth-inhibitory effects of ISIR-042 and bortezomib were not associated with caspase-10 which modulates autophagic response for survival. Expression of autophagic markers such as LC3 and p62 was also examined. ISIR-042 exerted modest effects in the presence or absence of bortezomib. ER stress-mediated CHOP induction promotes the cytotoxic effects of proteasome inhibitors on many cancer cells. Carfilzomib and ISIR-042 cotreatment significantly induced CHOP mRNA expression, suggesting that ISIR-042 and proteasome inhibitor cotreatment induced apoptosis by enhancing ER stress and activating CHOP. This was confirmed using CHOP-/- MEF cells. Our results showed that carfilzomib and ISIR-042 cotreatment did not exert cytotoxic effects on CHOP-/- MEF cells. These results suggest that treatment of multiple myelomas with ISIR-042-supplemented proteasome inhibitor-based chemotherapy exerts beneficial anticancer effects. Disclosures Suzuki: Chugai: Honoraria; Bristol-Myers Squibb: Honoraria; Kyowa Hakko kirin: Honoraria. Suzumiya:Astellas: Research Funding; Takeda: Honoraria; Eisai: Honoraria, Research Funding; Kyowa Hakko kirin: Research Funding; Chugai: Honoraria, Research Funding; Toyama Chemical: Research Funding.

Using muscle gene expression to estimate triacylglyceride deposition, and relative contributions of fatty acid synthesis and fatty acid import in intramuscular fat in cattle

2014 ◽  
Vol 54 (9) ◽  
pp. 1436 ◽  
Author(s):  
B. P. Dalrymple ◽  
B. Guo ◽  
G. H. Zhou ◽  
W. Zhang
Keyword(s):  
Gene Expression ◽  
Fatty Acid ◽  
De Novo ◽  
Acid Synthesis ◽  
Intramuscular Fat ◽  
Muscle Gene Expression ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Muscle Gene ◽  
The Impact

Intramuscular fat content (IMF%) in cattle influences the value of individual animals, especially for higher marbling markets. IMF is triacylglyceride (TAG) in lipid droplets in the intramuscular adipocytes. However, there are many different pathways from feed intake to the final common process of TAG synthesis and storage as IMF. To evaluate the relative importance of different pathways we compared changes in the expression of genes encoding proteins involved in the TAG and fatty acid (FA) synthesis pathways in the longissimus muscle of Piedmontese × Hereford (P×H) and Wagyu × Hereford (W×H) crosses. Based on these changes we have estimated the relative contributions of FA synthesised de novo in the intramuscular adipocyte and the uptake of circulating FA (both free and from TAG), from the diet or synthesised de novo in other tissues, to TAG deposition as IMF. We have analysed the impact of different developmental times and different diets on these processes. Increased de novo FA synthesis in intramuscular adipocytes appeared to contribute more than increased FA uptake from circulation to the additional TAG deposition in W×H compared with P×H cattle between 12 and 25 months (forage diet). Changing diet from forage to concentrate appeared to increase the importance of FA uptake from circulation relative to de novo FA synthesis for TAG synthesis in intramuscular adipocytes. These results are consistent with the literature based on analysis of lipid composition. Gene expression appears to provide a simple assay for identification of the source of FA for the deposition of IMF.

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