Search results for: Lymphocyte proliferation

Authors: Nurliyani, Eni Harmayani, Marsetyawan HNE Soesatyo

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Goat milk has an hypoallergenic effects, and allergic diseases related to abnormal of intestinal flora. Probiotic microorganisms do exert an activity on the immune system in the skin of the individual.The purpose of this study are to determine the number of leukocyte and lymphocyte proliferation in rat supplemented with fermented goat milk (acidophilus milk and kefir) and sensitized with dinitrochlorobenzene (DNCB). Female Wistar rats 6-8 weeks olds were divided into 3 treatment groups. The first group supplemented goat milk kefir, second group acidophilus goat milk, and third group as control. During 28-day experiment, on day 15 rat sensitized with allergen DNCB on the dorsal of the body, and on day 24 was challenged with DNCB on the ear. Sampling of blood and tissue of intestinal Peyer'patch (PP) were performed on day 14 (before DNCB sensitized) and on day 28 (after DNCB sensitized). The results showed the number of neutrophils in rats supplemented with acidophilus milk was higher (P<0.05) in after DNCB sensitized than before, but the lymphocyte count was lower. The number of monocytes, eosinophils, and basophils before and after DNCB sensitized have the same average for all treatments of milk fermented and control. Fermented goat milk (kefir and acidophilus milk) did not affect on rat PP lymphocyte proliferation culture supernatant, whereas the rat that had been DNCB sensitized showed higher in proliferative response to PHA mitogen (P <0.05) than before sensitized. In conclusion, supplementation of acidophilus goat milk with a dose of 2.0 ml / head / day on DNCB sensitized rat, can increase the number of neutrophils that play a role in innate immunity, however it was not able to increase lymphocyte proliferation that related to adaptive immunity.

Keywords: Leukocytes, Lymphocyte proliferation, Kefir, Acidophilus milk, Dinitrochlorobenzene

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Authors: Y. Baba hamed, A. Medjdoub, H. Merzouk, M. Narce

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The aim of this work was to study the in vitro effects of δ-lactam 1 and its 4-chlorophenyl derivative 2, on the proliferative responses of human lymphocytes and Th1 and Th2 cytokine secretion. The possible protective role of vitamin E on intracellular stress oxidative induced by these compounds was also investigated. Peripheral blood lymphocytes were isolated using differential centrifugation on a density gradient of Histopaque. They were cultured with mitogen concanavalin A, vitamin E (10 μM) and with different concentrations of the compounds 1 and 2 (0.1 to 10 μM). Proliferation (MTT assay), IL-2, INFγ and IL-4 (Elisa kits), intracellular superoxide anion were determined. 1 and 2 were immunostimulant and increased cytokine secretion with a shift away from Th1 response to Th2. These properties were however accompanied by an increase in intracellular oxidative stress. The presence of vitamin E exhibited protective effects by reducing δ- lactam-induced superoxide anion generation in lymphocytes.

Keywords: Cytokines, δ-Lactams, In vitro Lymphocyte Proliferation, Superoxide Anion

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Authors: Seyed Mazyar Mortazavi, Masod Memari, Hasan Ali Ahmadi, Zhaleh Abed

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VACTERL association is a rare disorder with various congenital malformations. The aetiology remains unknown. Combination of at least three congenital anomalies of the following criteria is required for diagnosis: vertebral defects, anal atresia, cardiac anomalies, tracheo-esophageal fistula, renal anomalies, and limb defects. The first case was 1-day old male neonate with multiple congenital anomalies was bore from 28 years old mother. The mother had history of pregnancy with lymphocyte therapy. His anomalies included: defects in thoracic and lumbar vertebral, anal atresia, bilateral hydronephrosis, atrial septal defect, and lower limb abnormality. Other anomalies were cryptorchidism and nasal canal narrowing. The second case was born with 32 weeks gestational age from mother with history of pregnancy with lymphocyte therapy. He had thoracic vertebral defect, cardiac anomalies and renal defect. diagnosis based on clinical finding is VACTERL association. Early diagnosis is very important to investigation and treatment of other coexistence anomalies. VACTERL association in mothers with history of pregnancy with lymphocyte therapy has suggested possibly of relationship between VACTERL association and this method of pregnancy.

Keywords: Anal atresia, tracheo-esophageal fistula, atrial septal defect, lymphocyte therapy.

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Authors: Duane Barker, John Holliday

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In a 10-week (May – August, 2008) Phase I trial, 840, 1+ rainbow trout, Oncorhynchus mykiss, received a commercial oral immunomodulator, Fin Immune™, at four different dosages (0, 10, 20 and 30 mg g-1) to evaluate immune response and growth. The overall objective of was to determine an optimal dosage of this product for rainbow trout that provides enhanced immunity with maximal growth and health. Biweekly blood samples were taken from 10 randomly selected fish in each tank (30 samples per treatment) to evaluate the duration of enhanced immunity conferred by Fin-Immune™. The immunological assessment included serum white blood cell (lymphocyte, neutrophil) densities and blood hematocrit (packed cell volume %). Of these three variables, only lymphocyte density increased significantly among trout fed Fin- Immune™ at 20 and 30 mg g-1 which peaked at week 6. At week 7, all trout were switched to regular feed (lacking Fin-Immune™) and by week 10, lymphocyte levels decreased among all levels but were still greater than at week 0. There was growth impairment at the highest dose of Fin-Immune™ tested (30 mg g-1) which can be associated with a physiological compensatory mechanism due to a dose-specific threshold level. Thus, our main objective of this Phase I study was achieved, the 20 mg g-1 dose of Fin-Immune™ should be the most efficacious (of those we tested) to use for a Phase II disease challenge trial.

Keywords: Blood Lymphocyte, Neutrophil Response of Cultured Rainbow Trout, Oncorhynchus mykiss, Oral Immunomodulator – 'Fin-ImmuneTM'.

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Authors: Forough Ataollahi, Belinda Pingguan-Murphy, Wan Abu Bakar Wan Abas, Noor Azuan Abu Osman

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Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al₂O₃) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al₂O₃ at curing temperature 50˚C for 4h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.

Keywords: Bovine aortic endothelial cells, extra cellular matrix, proliferation, stiffness.

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Authors: Mustafa M. Donma, Orkide Donma

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Obesity is defined as a severe chronic disease characterized by a low-grade inflammatory state. Therefore, inflammatory markers gained utmost importance during the evaluation of obesity and metabolic syndrome (MetS), a disease characterized by central obesity, elevated blood pressure, increased fasting blood glucose and elevated triglycerides or reduced high density lipoprotein cholesterol (HDL-C) values. Some inflammatory markers based upon complete blood cell count (CBC) are available. In this study, it was questioned which inflammatory marker was the best to evaluate the differences between various obesity groups. 514 pediatric individuals were recruited. 132 children with MetS, 155 morbid obese (MO), 90 obese (OB), 38 overweight (OW) and 99 children with normal BMI (N-BMI) were included into the scope of this study. Obesity groups were constituted using age- and sex-dependent body mass index (BMI) percentiles tabulated by World Health Organization. MetS components were determined to be able to specify children with MetS. CBC were determined using automated hematology analyzer. HDL-C analysis was performed. Using CBC parameters and HDL-C values, ratio markers of inflammation, which cover neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), monocyte-to-HDL-C ratio (MHR) were calculated. Statistical analyses were performed. The statistical significance degree was considered as p < 0.05. There was no statistically significant difference among the groups in terms of platelet count, neutrophil count, lymphocyte count, monocyte count, and NLR. PLR differed significantly between OW and N-BMI as well as MetS. Monocyte-to HDL-C value exhibited statistical significance between MetS and N-BMI, OB, and MO groups. HDL-C value differed between MetS and N-BMI, OW, OB, MO groups. MHR was the ratio, which exhibits the best performance among the other CBC-based inflammatory markers. On the other hand, when MHR was compared to HDL-C only, it was suggested that HDL-C has given much more valuable information. Therefore, this parameter still keeps its value from the diagnostic point of view. Our results suggest that MHR can be an inflammatory marker during the evaluation of pediatric MetS, but the predictive value of this parameter was not superior to HDL-C during the evaluation of obesity.

Keywords: Children, complete blood cell count, high density lipoprotein cholesterol, metabolic syndrome, obesity.

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Authors: M. Bastanfard, B. Jalaeian, S. Jafari

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Sonogram images of normal and lymphocyte thyroid tissues have considerable overlap which makes it difficult to interpret and distinguish. Classification from sonogram images of thyroid gland is tackled in semiautomatic way. While making manual diagnosis from images, some relevant information need not to be recognized by human visual system. Quantitative image analysis could be helpful to manual diagnostic process so far done by physician. Two classes are considered: normal tissue and chronic lymphocyte thyroid (Hashimoto's Thyroid). Data structure is analyzed using K-nearest-neighbors classification. This paper is mentioned that unlike the wavelet sub bands' energy, histograms and Haralick features are not appropriate to distinguish between normal tissue and Hashimoto's thyroid.

Keywords: Sonogram, thyroid, Haralick feature, wavelet.

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Authors: Jie Sun, Guoyou Yin, Xianqing Zhang, Qiusheng She, Zhaohui Xie, Lanying Chen, Anfang Zhao

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To assess the effects of functional protein on osteoblast, Large quantity of high-purity osteoblasts had been cultivated successfully by adopting sequential enzyme digestion. The growth curve of osteoblasts was protracted by cell counting. Proliferation of osteoblasts was assessed by MTT colorimetry. The experimental results show the functional protein can enhance proliferation, the properties of adhesion and discuss the effect of osteopontin on osteoblast.

Keywords: functional protein, osteoblast, MTT

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Authors: Mustafa M. Donma, Orkide Donma

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Obesity is a clinical state associated with low-grade inflammation. It is also a major risk factor for insulin resistance (IR). In its advanced stages, metabolic syndrome (MetS), a much more complicated disease which may lead to life-threatening problems, may develop. Obesity-mediated IR seems to correlate with the inflammation. Human studies performed particularly on pediatric population are scarce. The aim of this study is to detect possible associations between inflammation and IR in terms of some related ratios. 549 children were grouped according to their age- and sex-based body mass index (BMI) percentile tables of WHO. MetS components were determined. Informed consent and approval from the Ethics Committee for Clinical Investigations were obtained. The principles of the Declaration of Helsinki were followed. The exclusion criteria were infection, inflammation, chronic diseases and those under drug treatment. Anthropometric measurements were obtained. Complete blood cell, fasting blood glucose, insulin, and C-reactive protein (CRP) analyses were performed. Homeostasis model assessment of insulin resistance (HOMA-IR), systemic immune inflammation (SII) index, tense index, alanine aminotransferase to aspartate aminotransferase ratio (ALT/AST), neutrophils to lymphocyte (NLR), platelet to lymphocyte, and lymphocyte to monocyte ratios were calculated. Data were evaluated by statistical analyses. The degree for statistical significance was 0.05. Statistically significant differences were found among the BMI values of the groups (p < 0.001). Strong correlations were detected between the BMI and waist circumference (WC) values in all groups. Tense index values were also correlated with both BMI and WC values in all groups except overweight (OW) children. SII index values of children with normal BMI were significantly different from the values obtained in OW, obese, morbid obese and MetS groups. Among all the other lymphocyte ratios, NLR exhibited a similar profile. Both HOMA-IR and ALT/AST values displayed an increasing profile from N towards MetS3 group. BMI and WC values were correlated with HOMA-IR and ALT/AST. Both in morbid obese and MetS groups, significant correlations between CRP versus SII index as well as HOMA-IR versus ALT/AST were found. ALT/AST and HOMA-IR values were correlated with NLR in morbid obese group and with SII index in MetS group, (p < 0.05), respectively. In conclusion, these findings showed that some parameters may exhibit informative differences between the early and late stages of obesity. Important associations among HOMA-IR, ALT/AST, NLR and SII index have come to light in the morbid obese and MetS groups. This study introduced the SII index and NLR as important inflammatory markers for the discrimination of normal and obese children. Interesting links were observed between inflammation and IR in morbid obese children and those with MetS, both being late stages of obesity.

Keywords: Children, inflammation, insulin resistance, metabolic syndrome, obesity.

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Authors: Fawzi Irshaid, Fatiha Dilmi, Khaled Tarawneh, Raji Hadeth, Adnan Jaran, Ahad Al-Khatib

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P16/INK4A is tumor suppressor protein that plays a critical role in cell cycle regulation. Loss of P16 protein expression has been implicated in pathogenesis of many cancers, including lymphoma. Therefore, we sought to investigate if loss of P16 protein expression is associated with lymphoma and/or any specific lymphoma subtypes (Hodgkin-s lymphoma (HL) and nonHodgkin-s lymphoma (NHL)). Fifty-five lymphoma cases consisted of 30 cases of HL and 25 cases of NHL, with an age range of 3 to 78 years, were examined for loss of P16 by immunohistochemical technique using a specific antibody reacting against P16. In total, P16 loss was seen in 33% of all lymphoma cases. P16 loss was identified in 47.7% of HL cases. In contrast, only 16% of NHL showed loss of P16. Loss of P16 was seen in 67% of HL patients with 50 years of age or older, whereas P16 loss was found in only 42% of HL patients with less than 50 years of age. P16 loss in HL is somewhat higher in male (55%) than in female (30%). In subtypes of HL, P16 loss was found exclusively in all cases of lymphocyte depletion, lymphocyte predominance and unclassified cases, whereas P16 loss was seen in 39% of mixed cellularity and 29% of nodular sclerosis cases. In low grade NHL patients, P16 loss was seen in approximately one-third of cases, whereas no or very rare of P16 loss was found in intermediate and high grade cases. P16 loss did not show any correlation with age or gender of NHL patients. In conclusion, the high rate of P16 loss seen in our study suggests that loss of P16 expression plays a critical role in the pathogenesis of lymphoma, particularly with HL.

Keywords: B-cells, immunostaining, P16 protein, Reed-Sternberg cells, tumors.

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Authors: M. S. Hosseini, M. Tazzoli-Shadpour, I. Amjadi, A. A. Katbab, E. Jaefargholi-Rangraz

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Bionanotechnology deals with nanoscopic interactions between nanostructured materials and biological systems. Polymer nanocomposites with optimized biological activity have attracted great attention. Nanoclay is considered as reinforcing nanofiller in manufacturing of high performance nanocomposites. In current study, organomodified-nanoclay with negatively charged silicate layers was incorporated into biomedical grade silicone rubber. Nanoparticle loading has been tailored to enhance cell behavior. Addition of nanoparticles led to improved mechanical properties of substrate with enhanced strength and stiffness while no toxic effects was observed. Results indicated improved viability and proliferation of cells by addition of nanofillers. The improved mechanical properties of the matrix result in proper cell response through adjustment and arrangement of cytoskeletal fibers. Results can be applied in tissue engineering when enhanced substrates are required for improvement of cell behavior for in vivo applications.

Keywords: Biocompatibility, Composite, Organomodified- Nanoclay, Proliferation

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Authors: Mayada Alqaisi, Nasser Al-Shanti, Quiyu Wang, William S. Gilmore

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In-vitro mouse co-culture of E14 embryonic stem cells (ESCs) and OP9 stromal cells can recapitulate the earliest stages of haematopoietic development, not accessible in human embryos, supporting both haemogenic precursors and their primitive haematopoietic progeny. 1α, 25-Dihydroxy-vitamin D3 (VD3) has been demonstrated to be a powerful differentiation inducer for a wide variety of neoplastic cells, and could enhance early differentiation of ESCs into blood cells in E14/OP9 co-culture. This study aims to ascertain whether VD3 is key in promoting differentiation and suppressing proliferation, by separately investigating the effects of VD3 on the proliferation phase of the E14 cell line and on stromal OP9 cells.The results showed that VD3 inhibited the proliferation of the cells in a dose-dependent manner, quantitatively by decreased cell number, and qualitatively by alkaline-phosphatase staining that revealed significant differences between VD3-treated and untreated cells, characterised by decreased enzyme expression (colourless cells). Propidium-iodide cell-cycle analyses showed no significant percentage change in VD3-treated E14 and OP9 cells within their G and S-phases, compared to the untreated controls, despite the increased percentage of G-phase compared to the S-phase in a dosedependent manner. These results with E14 and OP9 cells indicate that adequate VD3 concentration enhances cellular differentiation and inhibits proliferation. The results also suggest that if E14 and OP9 cells were co-cultured andVD3-treated, there would be furtherenhanced differentiation of ESCs into blood cells.

Keywords: Differentiation, embryonic stem cells, OP9 stromal cells, 1α, 25-dihydroxy-vitamin D3

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Authors: Mustafa M. Donma, Orkide Donma

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Obesity is a low-grade inflammatory state. Childhood obesity is a multisystem disease, which is associated with a number of complications as well as potentially negative consequences. Gender is an important universal risk factor for many diseases. Hematological indices differ significantly by gender. This should be considered during the evaluation of obese children. The aim of this study is to detect hematologic indices that differ by gender in morbid obese (MO) children. A total of 134 MO children took part in this study. The parents filled an informed consent form and the approval from the Ethics Committee of Namik Kemal University was obtained. Subjects were divided into two groups based on their genders (64 females aged 10.2±3.1 years and 70 males aged 9.8±2.2 years; p ≥ 0.05). Waist-to-hip as well as head-to-neck ratios and body mass index (BMI) values were calculated. The children, whose WHO BMI-for age and sex percentile values were > 99 percentile, were defined as MO. Hematological parameters [haemoglobin, hematocrit, erythrocyte count, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red blood cell distribution width, leukocyte count, neutrophil %, lymphocyte %, monocyte %, eosinophil %, basophil %, platelet count, platelet distribution width, mean platelet volume] were determined by the automatic hematology analyzer. SPSS was used for statistical analyses. P ≤ 0.05 was the degree for statistical significance. The groups included children having mean±SD value of BMI as 26.9±3.4 kg/m² for males and 27.7±4.4 kg/m² for females (p ≥ 0.05). There was no significant difference between ages of females and males (p ≥ 0.05). Males had significantly increased waist-to-hip ratios (0.95±0.08 vs 0.91±0.08; p=0.005) and mean corpuscular hemoglobin concentration values (33.6±0.92 vs 33.1±0.83; p=0.001) compared to those of females. Significantly elevated neutrophil (4.69±1.59 vs 4.02±1.42; p=0.011) and neutrophil-to-lymphocyte ratios (1.70±0.71 vs 1.39±0.48; p=0.004) were detected in females. There was no statistically significant difference between groups in terms of C-reactive protein values (p ≥ 0.05). Adipose tissue plays important roles during the development of obesity and associated diseases such as metabolic syndrom and cardiovascular diseases (CVDs). These diseases may cause changes in complete blood cell count parameters. These alterations are even more important during childhood. Significant gender effects on the changes of neutrophils, one of the white blood cell subsets, were observed. The findings of the study demonstrate the importance of considering gender in clinical studies. The males and females may have distinct leukocyte-trafficking profiles in inflammation. Female children had more circulating neutrophils, which may be the indicator of an increased risk of CVDs, than male children within this age range during the late stage of obesity. In recent years, females represent about half of deaths from CVDs; therefore, our findings may be the indicator of the increasing tendency of this risk in females starting from childhood.

Keywords: Children, gender, morbid obesity, neutrophil-to-lymphocyte ratio.

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Authors: Morteza Elsa, Amirhossein Moghanian

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The major aim of this study was to evaluate the effect of CaO content on in vitro hydroxyapatite formation, MC3T3 cells cytotoxicity and proliferation as well as antibacterial efficiency of sol-gel derived SiO₂–CaO–P₂O₅ ternary system. For this purpose, first two grades of bioactive glass (BG); BG-58s (mol%: 60%SiO₂–36%CaO–4%P₂O₅) and BG-68s (mol%: 70%SiO₂–26%CaO–4%P₂O₅)) were synthesized by sol-gel method. Second, the effect of CaO content in their composition on in vitro bioactivity was investigated by soaking the BG-58s and BG-68s powders in simulated body fluid (SBF) for time periods up to 14 days and followed by characterization inductively coupled plasma atomic emission spectrometry (ICP-AES), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM) techniques. Additionally, live/dead staining, 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and alkaline phosphatase (ALP) activity assays were conducted respectively, as qualitatively and quantitatively assess for cell viability, proliferation and differentiations of MC3T3 cells in presence of 58s and 68s BGs. Results showed that BG-58s with higher CaO content showed higher in vitro bioactivity with respect to BG-68s. Moreover, the dissolution rate was inversely proportional to oxygen density of the BG. Live/dead assay revealed that both 58s and 68s increased the mean number live cells which were in good accordance with MTT assay. Furthermore, BG-58s showed more potential antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) bacteria. Taken together, BG-58s with enhanced MC3T3 cells proliferation and ALP activity, acceptable bioactivity and significant high antibacterial effect against MRSA bacteria is suggested as a suitable candidate in order to further functionalizing for delivery of therapeutic ions and growth factors in bone tissue engineering.

Keywords: Antibacterial, bioactive glass, hydroxyapatite, proliferation, sol-gel processes.

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Authors: Faten A. Khorshid

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Breast cancer is the most common malignancy in the world among women. Many therapies have been designed to treat this disease. Mamectomy, chemotherapy and radiotherapy are still the main therapies of breast cancer. However, the results were unsatisfactory and still far from the ideal treatment. PM 701is a natural product, has anticancer activity. The bioactive fraction PMF and subfraction PMFK had been isolated from PM701. PM 701 and its fractions were proved to have a cytotoxic properties against different cancer cell lines. This article is directed for the further examination of lyophilized PM701 and its active fractions on the growth of breast cancer cells (MCF-7). PM 701, PMF or PMFK were adding to the cultural medium, where MCF-7 is incubated. PM 701, PMF or PMFK were able to inhibit significantly the proliferation of MCF-7 cells, Moreover these new agents were proved to induce apoptosis of the breast cancer cells; through its direct effect on the nuclei.

Keywords: Anticancer agent, breast carcinoma, MCF-7 cell line, PM 701

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Authors: Mehrnaz Aminitabar, Moghan Amirhosseinian, Morteza Elsa

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Bioactive glasses (BGs) are a group of surface-reactive biomaterials used in clinical applications as implants or filler materials in the human body to repair and replace diseased or damaged bone. Sol-gel technique was employed to prepare a SiO₂-CaO-P₂O₅ glass with nominal composition of 58S BG with the addition of Sr and Li modifiers which imparts special properties to the BG. The effect of simultaneous addition of Sr and Li on bioactivity and biocompatibility, proliferation, alkaline phosphatase (ALP) activity of osteoblast cell line MC3T3-E1 and antibacterial property against methicillin-resistant Staphylococcus aureus (MRSA) bacteria were examined. BGs were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy before and after soaking the samples in the simulated body fluid (SBF) for different time intervals to characterize the formation of hydroxyapatite (HA) formed on the surface of BGs. Structural characterization indicated that the simultaneous presence of 5% Sr and 5% Li in 58S-BG composition not only did not retard HA formation because of opposite effect of Sr and Li of the dissolution of BG in the SBF but also, stimulated the differentiation and proliferation of MC3T3-E1s. Moreover, the presence of Sr and Li on dissolution of the ions resulted in an increase in the mean number of DAPI-labeled nuclei which was in good agreement with live/dead assay. The result of antibacterial tests revealed that Sr and Li-substituted 58S BG exhibited a potential antibacterial effect against MRSA bacteria. Because of optimal proliferation and ALP activity of MC3T3-E1cells, proper bioactivity and high antibacterial potential against MRSA, BG-5/5 is suggested as a multifunctional candidate for bone tissue engineering.

Keywords: Antibacterial activity, bioactive glass, sol-gel, strontium.

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Authors: A. Rohana, A. M., Fadzilah Adibah, M. S. Muhammad Roji

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Oleic acid (C18:1) play an important role in proliferation of fat cells. In this study, the effect of oleate on cells viability in 3T3-L1 cells (fat cells) was investigated. The 3T3-L1 cells were treated with various concentrations of oleate in the presence of 23 mM glucose. Oleate was added to adipogenic media (day 0) to investigate the influence of oleate on proliferation of postconfluent preadipocytes after 24 h induction. 0.1 mM oleate promoted cell division by increasing 33.9% number of cells from basal control in postconfluent preadipocytes. However, there were no significantly different in cells viability with control cells when oleate concentrations were increased up to 0.5 mM. When added to differentiated adipocytes (day 12) for 48 h, the number of cells decreased as oleate concentrations increased. 92.7% of cells lost demonstrated apoptosis and necrosis after 48 h with 0.5 mM oleate. The fluorochrome staining was examined under fluorescence microscopy using acridine orange and ethidium bromide double staining. Furthermore, the presence of high lactate (60.6% increased from basal control) released into plasma has shown the direct cytotoxicity of 0.5 mM oleate on adipocytes.

Keywords: adipocytes, apoptosis, oleate, postconfluentpreadipocytes

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Authors: Z. Hormozi Moghaddam, M. Mokhtari-Dizaji, M. Movahedin, M. E. Ravari

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Mechanical index (MI) is used for quantifying acoustic cavitation and the relationship between acoustic pressure and the frequency. In this study, modeling of the MI was applied to provide treatment protocol and to understand the effective physical processes on reproducibility of stem cells. The acoustic pressure and MI equations are modeled and solved to estimate optimal MI for 28, 40, 150 kHz and 1 MHz frequencies. Radial and axial acoustic pressure distribution was extracted. To validate the results of the modeling, the acoustic pressure in the water and near field depth was measured by a piston hydrophone. Results of modeling and experiments show that the model is consistent well to experimental results with 0.91 and 0.90 correlation of coefficient (p<0.05) for 1 MHz and 40 kHz. Low intensity ultrasound with 0.40 MI is more effective on the proliferation rate of the spermatogonial stem cells during the seven days of culture, in contrast, high MI has a harmful effect on the spermatogonial stem cells. This model provides proper treatment planning in vitro and in vivo by estimating the cavitation phenomenon.

Keywords: Ultrasound, mechanical index, modeling, stem cell.

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Authors: A. N. Gornostaeva, P. I. Bobyleva, E. R. Andreeva, L. B. Buravkova

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Multipotent mesenchymal stromal cells (MSCs) possess immunomodulatory properties. The effect of MSCs on the crucial cellular immunity compartment – T-cells is of a special interest. It is known that MSC tissue niche and expected milieu of their interaction with T- cells are characterized by low oxygen concentration, whereas the in vitro experiments usually are carried out at a much higher ambient oxygen (20%). We firstly evaluated immunomodulatory effects of MSCs on T-cells at tissue-related oxygen (5%) after interaction implied cell-to-cell contacts and paracrine factors only. It turned out that MSCs under reduced oxygen can effectively suppress the activation and proliferation of PHAstimulated T-cells and can provoke decrease in the production of proinflammatory and increase in anti-inflammatory cytokines. In hypoxia some effects were amplified (inhibition of proliferation, antiinflammatory cytokine profile shift). This impact was more evident after direct cell-to-cell interaction; lack of intercellular contacts could revoke the potentiating effect of hypoxia.

Keywords: Cell-to-cell interaction, low oxygen, MSC immunosuppression, T-cells.

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Authors: Mustafa M. Donma, Orkide Donma

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Obesity in children particularly draws attention, because it may threaten the individual’s future life due to many chronic diseases it may lead to. Most of these diseases including obesity itself altogether are related to inflammation. For this reason, inflammation-related parameters gain importance. Within this context, complete blood cell counts, ratios or indices derived from these counts have recently found some platform to be used as inflammatory markers. So far, mostly adipokines were investigated within the field of obesity. Metabolic inflammation is closely associated with cellular dysfunction. In this study, hematologic inflammatory markers and cytokines produced predominantly by the liver (fibroblast growth factor-21 (FGF-21) and fetuin A) were investigated in pediatric obesity. Two groups were constituted from 76 obese children based on World Health Organization criteria. Group 1 was composed of children, whose age- and sex-adjusted body mass index (BMI) percentiles were between 95 and 99. Group 2 consists of children, who are above 99th percentile. The first and the latter groups were defined as obese (OB) and morbid obese (MO). Anthropometric measurements of the children were performed. Informed consent forms and the approval of the institutional ethics committee were obtained. Blood cell counts and ratios were determined by automated hematology analyzer. The related ratios and indexes were calculated. Statistical evaluation of the data was performed by SPSS program. There was no statistically significant difference in terms of neutrophil-to lymphocyte ratio, monocyte-to-high density lipoprotein cholesterol ratio and platelet-to-lymphocyte ratio between the groups. Mean platelet volume and platelet distribution width values were decreased (p < 0.05), total platelet count, red cell distribution width (RDW) and systemic immune inflammation index values were increased (p < 0.01) in MO group. Both hepatokines were increased in the same group, however increases were not statistically significant. In this group, also a strong correlation was calculated between FGF-21 and RDW when controlled by age, hematocrit, iron and ferritin (r = 0.425; p < 0.01). In conclusion, the association between RDW, a hematologic inflammatory marker, and FGF-21, an inflammation-related hepatokine, found in MO group is an important finding discriminating between OB and MO children. This association is even more powerful when controlled by age and iron-related parameters.

Keywords: Childhood obesity, fetuin A, fibroblast growth factor-21, hematologic markers, red cell distribution width.

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Authors: Ibrahim Mohammed Saeed Shnawa

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The avian phytohaemagglutinin skin test is being proved as an in vivo system for the evaluation an avian in vivo T cell mitogenicity. The test system was one week old Gallus domesticus broiler Chickens. Five replicates were done for each of the whole, 1:10 dilutions of each of 0.05 IU tuberculin, tetanus immunoglobulin and DPT vaccine as test materials. The evaluation parameters were the skin indurations and lymphoblast percentages in bone marrow lymphocytes. Tuberculin indurations were 2.06 and 1.26mm for 0.05 IU respectively while lymphoblast percent were 0.234 and 0.1 accordingly. The skin indurations of 135mg/ml and 1.35mg/ml tetanus immunoglobulin were 4.86 and 3.96mm while lymphoblast percentages were 0.3 and 0.14 respectively. The whole DPT and 1:10 concentration were with 4.5 and 3.2mm while their lymphoblast percentages were 0.28 and 0.12 accordingly. Thus the mitogenicity of the test materials was of dependant type.

Keywords: DPT, Mitogenicity, Tetenus, immunoglobulin, Tubercular.

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Authors: Kiranmai S.Rai

Abstract:

The neurogenic potential of many herbal extracts used in Indian medicine is hitherto unknown. Extracts derived from Clitoria ternatea Linn have been used in Indian Ayurvedic system of medicine as an ingredient of “Medhya rasayana", consumed for improving memory and longevity in humans and also in treatment of various neurological disorders. Our earlier experimental studies with oral intubation of Clitoria ternatea aqueous root extract (CTR) had shown significant enhancement of learning and memory in postnatal and young adult Wistar rats. The present study was designed to elucidate the in vitro effects of 200ng/ml of CTR on proliferation, differentiation and growth of anterior subventricular zone neural stem cells (aSVZ NSC-s) derived from prenatal and postnatal rat pups. Results show significant increase in proliferation and growth of neurospheres and increase in the yield of differentiated neurons of aSVZ neural precursor cells (aSVZNPC-s) at 7 days in vitro when treated with 200ng/ml of CTR as compared to age matched control. Results indicate that CTR has growth promoting neurogenic effect on aSVZ neural stem cells and their survival similar to neurotrophic factors like Survivin, Neuregulin 1, FGF-2, BDNF possibly the basis for enhanced learning and memory.

Keywords: Anterior subventricular zone (aSVZ) neural stemcell, Clitoria ternatea, Learning and memory, Neurogenesis.

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Authors: S. A. H. Feghhi, Z. Gholamzadeh, Z. Alipoor, C. Tenreiro

Abstract:

Currently, thorium fuel has been especially noticed because of its proliferation resistance than long half-life alpha emitter minor actinides, breeding capability in fast and thermal neutron flux and mono-isotopic naturally abundant. In recent years, efficiency of minor actinide burning up in PWRs has been investigated. Hence, a minor actinide-contained thorium based fuel matrix can confront both proliferation resistance and nuclear waste depletion aims. In the present work, minor actinide depletion rate in a CANDU-type nuclear core modeled using MCNP code has been investigated. The obtained effects of minor actinide load as mixture of thorium fuel matrix on the core neutronics has been studied with comparing presence and non-presence of minor actinide component in the fuel matrix. Depletion rate of minor actinides in the MA-contained fuel has been calculated using different power loads. According to the obtained computational data, minor actinide loading in the modeled core results in more negative reactivity coefficients. The MA-contained fuel achieves less radial peaking factor in the modeled core. The obtained computational results showed 140 kg of 464 kg initial load of minor actinide has been depleted in during a 6-year burn up in 10 MW power.

Keywords: Minor actinide burning, CANDU-type reactor, MCNPX code, Neutronic parameters.

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Authors: N. Khatiashvili, Ch. Pirumova, V. Akhobadze

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In the paper the mathematical model of tumor growth is considered. New capillary network formation, which supply cancer cells with the nutrients, is taken into the account. A formula estimating a tumor growth in connection with the number of capillaries is obtained.

Keywords: Differential Equations, Mathematical Models, Vascular Endothelial, Tumor

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Authors: Elhamalsadat Ghaffari, Moghan Amirhosseinian, Amir Khaleghipour

Abstract:

Multifunctional bioactive glasses (BGs) are designed with a focus on the provision of bactericidal and biological properties desired for angiogenesis, osteogenesis, and ultimately potential applications in bone tissue engineering. To achieve these, six sol-gel copper/magnesium substituted derivatives of 58S-BG, i.e. a mol% series of 60SiO₂-4P₂O₅-5CuO-(31-x) CaO/xMgO (where x=0, 1, 3, 5, 8, and 10), were synthesized. Afterwards, the effect of MgO/CaO substitution on the in vitro formation of nano-hydroxyapatite (HA), osteoblast-like cell responses and BGs antibacterial performance were studied. During the BGs synthesis, the elimination of nitrates was achieved at 700 °C that prevented the BGs crystallization and stabilized the obtained dried gels. The structural and morphological evaluations were performed with X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). These characterizations revealed that Cu-substituted 58S-BG consisting of 5 mol% MgO (BG-5/5) slightly had retarded the formation of HA. In addition, Cu-substituted 58S-BGs consisting 8 mol% and 10 mol% MgO (BG-5/8 and BG-5/10) displayed lower bioactivity probably due to the lower ion release rate of Ca–Si into the simulated body fluid (SBF). The determination of 3-(4, 5 dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and alkaline phosphate (ALP) activities proved that the highest values of both differentiation and proliferation of MC3T3-E1 cells can be obtained from a 5 mol% MgO substituted BG, while the over addition of MgO (8 mol% and 10 mol%) decreased the bioactivity. Furthermore, these novel Cu/Mg-substituted 58S-BGs displayed antibacterial effect against methicillin-resistant Staphylococcus aureus bacteria. Taken together, the results suggest the equally-substituted BG-5/5 (i.e. the one consists of 5 mol% of both CuO and MgO) as a promising candidate for bone tissue engineering, among all newly designed BGs in this work, owing to its desirable cell proliferation, ALP activity and antibacterial properties.

Keywords: Apatite, bioactivity, biomedical applications sol-gel processes.

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Authors: Chih-Wei Chao, Jiashing Yu

Abstract:

To mimic the natural circumstances of cell growth in an organism, we present three-dimensional (3D) scaffolds fabricated by microfluidics for cultivation. This work investigates the cellular behaviors of rat cardiomyocytes in gelatin 3D scaffolds compared to those on 2D control, such as proliferation, viability and morphology. We found that the scaffolds may induce skeletal differentiation of H9c2 cells.

Keywords: Microfluidic device, H9c2, tissue engineering, 3D scaffolds.

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Authors: Ali Asghar Saki, Sayed Ali Hosseini Siyar, Abbass Ashoori

Abstract:

This study was conducted to evaluate the effect of Echinacea purpurea on the expression of cyclooxygenase-2 (COX-2), interleukin-17F (IL-17F) in seven-day-old broiler chickens. Four groups were fed with concentration of 0 g/kg, 5 g/kg, 10 g/kg and 20 g/kg from the root of E. purpurea in the basal diet and two other groups were only fed with the basal diet for 21 days. At the 28^th day, lipopolysaccharide (LPS, 2 mg/kg diet) was injected in four groups and the basal diet group was injected by saline as control. The chickens’ spleen RNA expression was measured for the COX-2 and IL-17F genes by Real-Time PCR. The results have shown that chickens which were fed E. purpurea had a lower COX-2 and IL-17F mRNA expression. The chickens who have received LPS only, lymphocyte was lower than other treatments. Vital organ weights were not significantly different, but body weight loss was recovered by dietary herbs inclusion. The results of this study have shown the positive effect of an anti-inflammatory herb to prevent the undesirable effect of inflammation.

Keywords: Echinacea purpurea, broiler chickens, gene expression, lipopolysaccharide.

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Authors: Hazim Saadoon Aljewari, Mohammed Ibraheem Nader, Abdul Hussain M. Alfaisal, NatthidaWeerapreeyakul, Sahapat

Abstract:

L-asparaginase was extracted from pathogenic Escherichia coli which was isolated from urinary tract infection patients. L-asparaginase was purified 96-fold by ultrafiltration, ion exchange and gel filtration giving 39.19% yield with final specific activity of 178.57 IU/mg. L-asparaginase showed 138,356±1,000 Dalton molecular weight with 31024±100 Dalton molecular mass. Kinetic properties of enzyme resulting 1.25×10-5 mM Km and 2.5×10-3 M/min Vmax. L-asparaginase showed a maximum activity at pH 7.5 when incubated at 37 ºC for 30 min and illustrated its full activity (100%) after 15 min incubation at 20-37 ºC, while 70% of its activity was lost when incubated at 60 ºC. L-asparaginase showed cytotoxicity to U937 cell line with IC50 0.5±0.19 IU/ml, and selectivity index (SI=7.6) about 8 time higher selectivity over the lymphocyte cells. Therefore, the local pathogenic E. coli strains may be used as a source of high yield of L-asparaginase to produce anti cancer agent with high selectivity.

Keywords: L-asparaginase, Purification, Cytotoxicity, selectivity index

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Authors: N. Hussain, G. Jaffery, A.N. Sabri, S. Hasnain

Abstract:

The main aim is to perform mutational analysis of CTLA4 gene Exon 1 in SLE patients. A total of 61 SLE patients fulfilling “American College of Rheumatology (ACR) criteria" and 61 controls were enrolled in this study. The region of CTLA4 gene exon 1 was amplified by using Step-down PCR technique. Extracted DNA of band 354 bp was sequenced to analyze mutations in the exon-1 of CTLA-4 gene. Further, protein sequences were identified from nucleotide sequences of CTLA4 Exon 1 by using Expasy software and through Blast P software it was found that CTLA4 protein sequences of Pakistani SLE patients were similar to that of Chinese SLE population. No variations were found after patients sequences were compared with that of the control sequence. Furthermore it was found that CTLA4 protein sequences of Pakistani SLE patients were similar to that of Chinese SLE population. Thus CTLA4 gene may not be responsible for an autoimmune disease SLE.

Keywords: American College of Rheumatology criteria, autoimmune disease, Cytotoxic T Lymphocyte Antigen-4, Polymerase Chain Reaction, Systemic Lupus Erythematosus

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Authors: Alireza Arab, Morteza Elsa, Amirhossein Moghanian

Abstract:

In this study, experiments were carried out to achieve a promising multifunctional and modified silicate based bioactive glass (BG). The main aim of the study was investigating the effect of lithium (Li) and magnesium (Mg) substitution, on in vitro bioactivity of substituted-58S BG. Moreover, it is noteworthy to state that modified BGs were synthesized in 60SiO₂–(36-x)CaO–4P₂O₅–(x)Li₂O and 60SiO₂–(36-x)CaO–4P₂O₅–(x)MgO (where x = 0, 5, 10 mol.%) quaternary systems, by sol-gel method. Their performance was investigated through different aspects such as biocompatibility, antibacterial activity as well as their effect on alkaline phosphatase (ALP) activity, and proliferation of MC3T3 cells. The antibacterial efficiency was evaluated against methicillin-resistant Staphylococcus aureus bacteria. To do so, CaO was substituted with Li₂O and MgO up to 10 mol % in 58S-BGs and then samples were immersed in simulated body fluid up to 14 days and then, characterized by X-ray diffraction, Fourier transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometry, and scanning electron microscopy. Results indicated that this modification led to a retarding effect on in vitro hydroxyapatite (HA) formation due to the lower supersaturation degree for nucleation of HA compared with 58s-BG. Meanwhile, magnesium revealed further pronounced effect. The 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) and ALP analysis illustrated that substitutions of both Li₂O and MgO, up to 5 mol %, had increasing effect on biocompatibility and stimulating proliferation of the pre-osteoblast MC3T3 cells in comparison to the control specimen. Regarding to bactericidal efficiency, the substitution of either Li or Mg for Ca in the 58s BG composition led to statistically significant difference in antibacterial behaviors of substituted-BGs. Meanwhile, the sample containing 5 mol % CaO/Li₂O substitution (BG-5L) was selected as a multifunctional biomaterial in bone repair/regeneration due to the improved biocompatibility, enhanced ALP activity and antibacterial efficiency among all of the synthesized L-BGs and M-BGs.

Keywords: Alkaline, alkaline earth, bioactivity, biomedical applications, sol-gel processes.

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