Search results for: phenotypic markers
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 848

Search results for: phenotypic markers

638 Study of Circulatory MiR-122 and MiR-130a Expression among Chronic Hepatitis C Egyptian Patients

Authors: Hend K. Moosa, Eman A. Rashwan, Ezzat M. Hassan, Amany A. Ghazy, Amel G. Sheredy

Abstract:

The stability of microRNA (miR) in the circulation can show a great progress toward the discovery of non-invasive diagnostic and prognostic biomarkers in many diseases. In the present study, circulatory miR-122 and miR-130a were analysed in chronic hepatitis C Egyptian patients in predicting the clinical outcome of interferon treatment. In addition, their expression levels were correlated to viral RNA levels, necro-inflammatory markers (AST, ALT) and to each other. This study was conducted on 51 subjects where 36 were chronic HCV patients in which they were divided into naive and interferon treated HCV patients (responders and non-responders) and 15 matched healthy controls. Serum quantification of miR-122 and miR-130a were performed by quantitative Real-time Polymerase Chain Reaction (qRT-PCR). The results showed a significant upregulation of miR-122 in non-responder patients (P=0.049). By receiver operating characteristic analysis curve, miR-122 revealed 65% sensitivity and 92.3% specificity in predicting non-responsiveness of patients to IFN treatment, while miR-130a showed a sensitivity of 100% and specificity of 53.85%. Remarkably, there was a significant positive correlation between miR-122 and miR-130a in naive HCV patients (r=0.714, p=0.003). However, there was no significant correlation between serum miR-122, miR-130a expression levels and necro-inflammatory markers (AST, ALT). To conclude, miR-122 and miR-130a have a significant association with viral RNA levels and accordingly, they may have a synergistic power in promoting viral replication. Interestingly, miR-122 and miR-130a have a predictive power in predicting clinical outcome of IFN treatment which can be further studied in currently used drugs in order to reduce the socio-economic burden of potentially non-responders.

Keywords: hepatitis C, microRNA, miR-122, miR-130a

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637 Paradigm Shift in Classical Drug Research: Challenges to Mordern Pharmaceutical Sciences

Authors: Riddhi Shukla, Rajeshri Patel, Prakruti Buch, Tejas Sharma, Mihir Raval, Navin Sheth

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Many classical drugs are claimed to have blood sugar lowering properties that make them valuable for people with or at high risk of type 2 diabetes. Vijaysar (Pterocarpus marsupium) and Gaumutra (Indian cow urine) both have been shown antidiabetic property since primordial time and both shows synergistic effect in combination for hypoglycaemic activity. The study was undertaken to investigate the hypoglycaemic and anti-diabetic effects of the combination of Vijaysar and Gaumutra which is a classical preparation mentioned in Ayurveda named as Pramehari ark. Rats with Type 2 diabetes which is induced by streptozotocin (STZ, 35mg/kg) given a high-fat diet for one month and compared with normal rats. Diabetic rats showed raised level of body weight, triglyceride (TG), total cholesterol, HDL, LDL, and D-glucose concentration and other serum, cardiac and hypertrophic parameters in comparison of normal rats. After treatment of different doses of drug the level of parameters like TG, total cholesterol, HDL, LDL, and D-glucose concentration found to be decreased in standard as well as in treatment groups. In addition treatment groups also found to be decreased in the level of serum markers, cardiac markers, and hypertrophic parameters. The findings demonstrated that Pramehari ark prevented the pathological progression of type 2 diabetes in rats.

Keywords: cow urine, hypoglycemic effect, synergic effect, type 2 diabetes, vijaysar

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636 Formulation and Anticancer Evaluation of Beta-Sitosterol in Henna Methanolic Extract Embedded in Controlled Release Nanocomposite

Authors: Sanjukta Badhai, Durga Barik, Bairagi C. Mallick

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In the present study, Beta-Sitosterol in Lawsonia methanolic leaf extract embedded in controlled release nanocomposite was prepared and evaluated for in vivo anticancer efficacy in dimethyl hydrazine (DMH) induced colon cancer. In the present study, colon cancer was induced by s.c injection of DMH (20 mg/kg b.wt) for 15 weeks. The animals were divided into five groups as follows control, DMH alone, DMH and Beta Sitosterol nanocomposite (50mg/kg), DMH and Beta Sitosterol nanocomposite (100 mg/kg) and DMH and Standard Silymarin (100mg/kg) and the treatment was carried out for 15 weeks. At the end of the study period, the blood was withdrawn, and serum was separated for haematological, biochemical analysis and tumor markers. Further, the colonic tissue was removed for the estimation of antioxidants and histopathological analysis. The results of the study displays that DMH intoxication elicits altered haematological parameters (RBC,WBC, and Hb), elevated lipid peroxidation and decreased antioxidants level (SOD, CAT, GPX, GST and GSH), elevated lipid profiles (cholesterol and triglycerides), tumor markers (CEA and AFP) and altered colonic tissue histology. Meanwhile, treatment with Beta Sitosterol nanocomposites significantly restored the altered biochemicals parameters in DMH induced colon cancer mediated by its anticancer efficacy. Further, Beta Sitosterol nanocomposite (100 mg/kg) showed marked efficacy.

Keywords: nanocomposites, herbal formulation, henna, beta sitosterol, colon cancer, dimethyl hydrazine, antioxidant, lipid peroxidation

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635 Genomics of Adaptation in the Sea

Authors: Agostinho Antunes

Abstract:

The completion of the human genome sequencing in 2003 opened a new perspective into the importance of whole genome sequencing projects, and currently multiple species are having their genomes completed sequenced, from simple organisms, such as bacteria, to more complex taxa, such as mammals. This voluminous sequencing data generated across multiple organisms provides also the framework to better understand the genetic makeup of such species and related ones, allowing to explore the genetic changes underlining the evolution of diverse phenotypic traits. Here, recent results from our group retrieved from comparative evolutionary genomic analyses of selected marine animal species will be considered to exemplify how gene novelty and gene enhancement by positive selection might have been determinant in the success of adaptive radiations into diverse habitats and lifestyles.

Keywords: marine genomics, evolutionary bioinformatics, human genome sequencing, genomic analyses

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634 Expression Profiling and Immunohistochemical Analysis of Squamous Cell Carcinoma of Head and Neck (Tumor, Transition Zone, Normal) by Whole Genome Scale Sequencing

Authors: Veronika Zivicova, Petr Broz, Zdenek Fik, Alzbeta Mifkova, Jan Plzak, Zdenek Cada, Herbert Kaltner, Jana Fialova Kucerova, Hans-Joachim Gabius, Karel Smetana Jr.

Abstract:

The possibility to determine genome-wide expression profiles of cells and tissues opens a new level of analysis in the quest to define dysregulation in malignancy and thus identify new tumor markers. Toward this long-term aim, we here address two issues on this level for head and neck cancer specimen: i) defining profiles in different regions, i.e. the tumor, the transition zone and normal control and ii) comparing complete data sets for seven individual patients. Special focus in the flanking immunohistochemical part is given to adhesion/growth-regulatory galectins that upregulate chemo- and cytokine expression in an NF-κB-dependent manner, to these regulators and to markers of differentiation, i.e. keratins. The detailed listing of up- and down-regulations, also available in printed form (1), not only served to unveil new candidates for testing as marker but also let the impact of the tumor in the transition zone become apparent. The extent of interindividual variation raises a strong cautionary note on assuming uniformity of regulatory events, to be noted when considering therapeutic implications. Thus, a combination of test targets (and a network analysis for galectins and their downstream effectors) is (are) advised prior to reaching conclusions on further perspectives.

Keywords: galectins, genome scale sequencing, squamous cell carcinoma, transition zone

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633 Effect of Online Mindfulness Training to Tertiary Students’ Mental Health: An Experimental Research

Authors: Abigaile Rose Mary R. Capay, Janne Ly Castillon-Gilpo, Sheila A. Javier

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The transition to online learning has been a challenging feat on the mental health of tertiary students. This study investigated whether learning mindfulness strategies online would help in improving students’ imagination, conscientiousness, extraversion, agreeableness and emotional stability, as measured by the International Personality Item Pool (IPIP) Big Five Factor Markers, as well as their dispositional mindfulness as measured by the Mindfulness Attention Awareness Scale (MAAS). Fifty-two college students participated in the experiment. The 23 participants assigned to the treatment condition received 6-weekly experiential sessions of online mindfulness training and were advised to follow a daily mindfulness practice, while the 29 participants from the control group only received a 1-hour lecture. Scores were collected at pretest and posttest. Findings show that there was a significant difference in the pretest and posttest scores of students assigned in the treatment group, likewise medium effect sizes in the variables: dispositional mindfulness (t (22) = 2.64, p = 0.015, d = .550), extraversion (t (22) = 2.76, p = 0.011, d = 0.575), emotional stability (t (22) = 2.99, p = 0.007, d = .624), conscientiousness (t (22) = 2.74, p = 0.012, d = .572) and imagination (t (22) = 4.08, p < .001), but not for agreeableness (t (22) = 2.01, p = 0.057, d = .419). No significant differences were observed on the scores of the control group. Educational institutions are recommended to consider teaching basic mindfulness strategies to tertiary students, as a valuable resource in improving their mental health as they navigate through adjustments in online learning.

Keywords: mindfulness, school-based interventions, MAAS, IPIP Big Five Markers, experiment

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632 Nitric Oxide and Blood Based Ratios as Promising Immuno-Markers in Patients with Complicated Crohn’s Disease: Benefits for Predicting Therapy Response

Authors: Imene Soufli, Abdelkrim Hablal, Manel Amri, Moussa Labsi, Rania Sihem Boussa, Nassim Sid Idris, Chafia Touil-Boukoffa

Abstract:

Crohn’s Disease (CD) is a relapsing–remitting inflammatory bowel disease with a progressive course. The aim of our study was to evaluate the relationship between the immunomarkers: Nitric Oxide (NO), pro-inflammatory cytokines, and blood count-based ratios and the outcome of corticosteroid or anti-TNF-α therapy in patients with complicated Crohn’s Disease. In this context, we evaluated the NLR as the ratio of neutrophil count to lymphocyte count, PLR as the ratio of platelet counts to lymphocyte count, and MLR as the ratio of monocyte count to lymphocyte count in patients and controls. Furthermore, we assessed NO production by the Griess method in plasma along with iNOS and NF-κB expression by immunofluorescence method in intestinal tissues of patients and controls. In the same way, we evaluated plasma TNF-α, IL-17A, and IL-10 levels using ELISA. Our results indicate that blood count-based ratios NLR, PLR, and MLR were significantly higher in patients compared to controls. In addition, increased systemic levels of NO, TNF-α, and IL-17A and colonic expression of iNOS and NF-κB were observed in the same patients. Interestingly, the high ratio of NLR and MLR, as well as NO production, was significantly decreased in treated patients. Collectively, our findings suggest that Nitric Oxide, as well as the blood count-based ratios (NLR, PLR, MLR), could constitute useful immuno-markers in complicated Crohn’s Disease, predicting the response to treatment

Keywords: complicated crohn’s disease, nitric oxide, blood count-based ratios, treatments, pro-inflammatory cytokines

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631 Platelet Volume Indices: Emerging Markers of Diabetic Thrombocytopathy

Authors: Mitakshara Sharma, S. K. Nema

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Diabetes mellitus (DM) is metabolic disorder prevalent in pandemic proportions, incurring significant morbidity and mortality due to associated vascular angiopathies. Platelet related thrombogenesis plays key role in pathogenesis of these complications. Most patients with type II DM suffer from preventable vascular complications and early diagnosis can help manage these successfully. These complications are attributed to platelet activation which can be recognised by the increase in Platelet Volume Indices(PVI) viz. Mean Platelet Volume(MPV) and Platelet Distribution Width(PDW). This study was undertaken with the aim of finding a relationship between PVI and vascular complications of Diabetes mellitus, their importance as a causal factor in these complications and use as markers for early detection of impending vascular complications in patients with poor glycaemic status. This is a cross-sectional study conducted for 2 years with total 930 subjects. The subjects were segregated in 03 groups on basis of glycosylated haemoglobin (HbA1C) as: - (a) Diabetic, (b) Non-Diabetic and (c) Subjects with Impaired fasting glucose(IFG) with 300 individuals in IFG and non-diabetic group & 330 individuals in diabetic group. The diabetic group was further divided into two groups: - (a) Diabetic subjects with diabetes related vascular complications (b) Diabetic subjects without diabetes related vascular complications. Samples for HbA1C and platelet indices were collected using Ethylene diamine tetracetic acid(EDTA) as anticoagulant and processed on SYSMEX-XS-800i autoanalyser. The study revealed stepwise increase in PVI from non-diabetics to IFG to diabetics. MPV and PDW of diabetics, IFG and non diabetics were 17.60 ± 2.04, 11.76 ± 0.73, 9.93 ± 0.64 and 19.17 ± 1.48, 15.49 ± 0.67, 10.59 ± 0.67 respectively with a significant p value 0.00 and a significant positive correlation (MPV-HbA1c r = 0.951; PDW-HbA1c r = 0.875). However, significant negative correlation was found between glycaemic levels and total platelet count (PC- HbA1c r =-0.164). MPV & PDW of subjects with and without diabetes related complications were (15.14 ± 1.04) fl & (17.51±0.39) fl and (18.96 ± 0.83) fl & (20.09 ± 0.98) fl respectively with a significant p value 0.00.The current study demonstrates raised platelet indices & reduced platelet counts in association with rising glycaemic levels and diabetes related vascular complications across various study groups & showed that platelet morphology is altered with increasing glycaemic levels. These changes can be known by measurements of PVI which are important, simple, cost effective, effortless tool & indicators of impending vascular complications in patients with deranged glycaemic control. PVI should be researched and explored further as surrogate markers to develop a clinical tool for early recognition of vascular changes related to diabetes and thereby help prevent them. They can prove to be more useful in developing countries with limited resources. This study is multi-parameter, comprehensive with adequately powered study design and represents pioneering effort in India on account of the fact that both Platelet indices (MPV & PDW) along with platelet count have been evaluated together for the first time in Diabetics, non diabetics, patients with IFG and also in the diabetic patients with and without diabetes related vascular complications.

Keywords: diabetes, HbA1C, IFG, MPV, PDW, PVI

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630 Liver and Liver Lesion Segmentation From Abdominal CT Scans

Authors: Belgherbi Aicha, Hadjidj Ismahen, Bessaid Abdelhafid

Abstract:

The interpretation of medical images benefits from anatomical and physiological priors to optimize computer- aided diagnosis applications. Segmentation of liver and liver lesion is regarded as a major primary step in computer aided diagnosis of liver diseases. Precise liver segmentation in abdominal CT images is one of the most important steps for the computer-aided diagnosis of liver pathology. In this papers, a semi- automated method for medical image data is presented for the liver and liver lesion segmentation data using mathematical morphology. Our algorithm is currency in two parts. In the first, we seek to determine the region of interest by applying the morphological filters to extract the liver. The second step consists to detect the liver lesion. In this task; we proposed a new method developed for the semi-automatic segmentation of the liver and hepatic lesions. Our proposed method is based on the anatomical information and mathematical morphology tools used in the image processing field. At first, we try to improve the quality of the original image and image gradient by applying the spatial filter followed by the morphological filters. The second step consists to calculate the internal and external markers of the liver and hepatic lesions. Thereafter we proceed to the liver and hepatic lesions segmentation by the watershed transform controlled by markers. The validation of the developed algorithm is done using several images. Obtained results show the good performances of our proposed algorithm

Keywords: anisotropic diffusion filter, CT images, hepatic lesion segmentation, Liver segmentation, morphological filter, the watershed algorithm

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629 Role of Zinc Adminstration in Improvement of Faltering Growth in Egyption Children at Risk of Environmental Enteric Dysfunction

Authors: Ghada Mahmoud El Kassas, Maged Atta El Wakeel

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Background: Environmental enteric dysfunction (EED) is impending trouble that flared up in the last decades to be pervasive in infants and children. EED is asymptomatic villous atrophy of the small bowel that is prevalent in the developing world and is associated with altered intestinal function and integrity. Evidence has suggested that supplementary zinc might ameliorate this damage by reducing gastrointestinal inflammation and may also benefit cognitive development. Objective: We tested whether zinc supplementation improves intestinal integrity, growth, and cognitive function in stunted children predicted to have EED. Methodology: This case–control prospective interventional study was conducted on 120 Egyptian Stunted children aged 1-10 years who recruited from the Nutrition clinic, the National research center, and 100 age and gender-matched healthy children as controls. At the primary phase of the study, Full history taking, clinical examination, and anthropometric measurements were done. Standard deviation score (SDS) for all measurements were calculated. Serum markers as Zonulin, Endotoxin core antibody (EndoCab), highly sensitive C-reactive protein (hsCRP), alpha1-acid glycoprotein (AGP), Tumor necrosis factor (TNF), and fecal markers such as myeloperoxidase (MPO), neopterin (NEO), and alpha-1-anti-trypsin (AAT) (as predictors of EED) were measured. Cognitive development was assessed (Bayley or Wechsler scores). Oral zinc at a dosage of 20 mg/d was supplemented to all cases and followed up for 6 months, after which the 2ry phase of the study included the previous clinical, laboratory, and cognitive assessment. Results: Serum and fecal inflammatory markers were significantly higher in cases compared to controls. Zonulin (P < 0.01), (EndoCab) (P < 0.001) and (AGP) (P < 0.03) markedly decreased in cases at the end of 2ry phase. Also (MPO), (NEO), and (AAT) showed a significant decline in cases at the end of the study (P < 0.001 for all). A significant increase in mid-upper arm circumference (MUAC) (P < 0.01), weight for age z-score, and skinfold thicknesses (P< 0.05 for both) was detected at end of the study, while height was not significantly affected. Cases also showed significant improvement of cognitive function at phase 2 of the study. Conclusion: Intestinal inflammatory state related to EED showed marked recovery after zinc supplementation. As a result, anthropometric and cognitive parameters showed obvious improvement with zinc supplementation.

Keywords: stunting, cognitive function, environmental enteric dysfunction, zinc

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628 The Value of Serum Procalcitonin in Patients with Acute Musculoskeletal Infections

Authors: Mustafa Al-Yaseen, Haider Mohammed Mahdi, Haider Ali Al–Zahid, Nazar S. Haddad

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Background: Early diagnosis of musculoskeletal infections is of vital importance to avoid devastating complications. There is no single laboratory marker which is sensitive and specific in diagnosing these infections accurately. White blood cell count, erythrocyte sedimentation rate, and C-reactive protein are not specific as they can also be elevated in conditions other than bacterial infections. Materials Culture and sensitivity is not a true gold standard due to its varied positivity rates. Serum Procalcitonin is one of the new laboratory markers for pyogenic infections. The objective of this study is to assess the value of PCT in the diagnosis of soft tissue, bone, and joint infections. Patients and Methods: Patients of all age groups (seventy-four patients) with a diagnosis of musculoskeletal infection are prospectively included in this study. All patients were subjected to White blood cell count, erythrocyte sedimentation rate, C-reactive protein, and serum Procalcitonin measurements. A healthy non infected outpatient group (twenty-two patients) taken as a control group and underwent the same evaluation steps as the study group. Results: The study group showed mean Procalcitonin levels of 1.3 ng/ml. Procalcitonin, at 0.5 ng/ml, was (42.6%) sensitive and (95.5%) specific in diagnosing of musculoskeletal infections with (positive predictive value of 87.5% and negative predictive value of 48.3%) and (positive likelihood ratio of 9.3 and negative likelihood ratio of 0.6). Conclusion: Serum Procalcitonin, at a cut – off of 0.5 ng/ml, is a specific but not sensitive marker in the diagnosis of musculoskeletal infections, and it can be used effectively to rule in the diagnosis of infection but not to rule out it.

Keywords: procalcitonin, infection, labratory markers, musculoskeletal

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627 Expression of Fibrogenesis Markers after Mesenchymal Stem Cells Therapy for Experimental Liver Cirrhosis

Authors: Tatsiana Ihnatovich, Darya Nizheharodava, Mikalai Halabarodzka, Tatsiana Savitskaya, Marina Zafranskaya

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Liver fibrosis is a complex of histological changes resulting from chronic liver disease accompanied by an excessive production and deposition of extracellular matrix components in the hepatic parenchyma. Liver fibrosis is a serious medical and social problem. Hepatic stellate cells (HSCs) make a significant contribution to the extracellular matrix deposition due to liver injury. Mesenchymal stem cells (MSCs) have a pronounced anti-inflammatory, regenerative and immunomodulatory effect; they are able to differentiate into hepatocytes and induce apoptosis of activated HSCs that opens the prospect of their use for preventing the excessive fibro-formation and the development of liver cirrhosis. The aim of the study is to evaluate the effect of MSCs therapy on the expression of fibrogenesis markers genes in liver tissue and HSCs cultures of rats with experimental liver cirrhosis (ELC). Materials and methods: ELC was induced by the common bile duct ligation (CBDL) in female Wistar rats (n = 19) with an average body weight of 250 (220 ÷ 270) g. Animals from the control group (n = 10) were sham-operated. On the 56th day after the CBDL, the rats of the experimental (n = 12) and the control (n = 5) groups received intraportal MSCs in concentration of 1×106 cells/animal (previously obtained from rat’s bone marrow) or saline, respectively. The animals were taken out of the experiment on the 21st day. HSCs were isolated by sequential liver perfusion in situ with following disaggregation, enzymatic treatment and centrifugation of cell suspension on a two-stage density gradient. The expression of collagen type I (Col1a1) and type III (Col3a1), matrix metalloproteinase type 2 (MMP2) and type 9 (MMP9), tissue inhibitor of matrix metalloproteinases type 1 (TIMP1), transforming growth factor β type 1 (TGFβ1) and type 3 (TGFβ3) was determined by real-time polymerase chain reaction. Statistical analysis was performed using Statistica 10.0. Results: In ELC rats compared to sham-operated animals, a significant increase of all studied markers expression was observed. The administration of MSCs led to a significant decrease of all detectable markers in the experimental group compared to rats without cell therapy. In ELC rats, an increased MMP9/TIMP1 ratio after cell therapy was also detected. The infusion of MSCs in the sham-operated animals did not lead to any changes. In the HSCs from ELC animals, the expression of Col1a1 and Col3a1 exceeded the similar parameters of the control group (p <0.05) and statistically decreased after the MSCs administration. The correlation between Col3a1 (Rs = 0.51, p <0.05), TGFβ1 (Rs = 0.6, p <0.01), and TGFβ3 (Rs = 0.75, p <0.001) expression in HSCs cultures and liver tissue has been found. Conclusion: Intraportal administration of MSCs to rats with ELC leads to a decreased Col1a1 and Col3a1, MMP2 and MMP9, TIMP1, TGFβ1 and TGFβ3 expression. The correlation between the expression of Col3a1, TGFβ1 and TGFβ3 in liver tissue and in HSCs cultures indicates the involvement of activated HSCs in the fibrogenesis that allows considering HSCs to be the main cell therapy target in ELC.

Keywords: cell therapy, experimental liver cirrhosis, hepatic stellate cells, mesenchymal stem cells

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626 Qf-Pcr as a Rapid Technique for Routine Prenatal Diagnosis of Fetal Aneuploidies

Authors: S. H. Atef

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Background: The most common chromosomal abnormalities identified at birth are aneuploidies of chromosome 21, 18, 13, X and Y. Prenatal diagnosis of fetal aneuploidies is routinely done by traditional cytogenetic culture, a major drawback of this technique is the long period of time required to reach a diagnosis. In this study, we evaluated the QF-PCR as a rapid technique for prenatal diagnosis of common aneuploidies. Method:This work was carried out on Sixty amniotic fluid samples taken from patients with one or more of the following indications: Advanced maternal age (3 case), abnormal biochemical markers (6 cases), abnormal ultrasound (12 cases) or previous history of abnormal child (39 cases).Each sample was tested by QF-PCR and traditional cytogenetic. Aneuploidy screenings were performed amplifying four STRs on chromosomes 21, 18, 13, two pseudoautosomal,one X linked, as well as the AMXY and SRY; markers were distributed in two multiplex QFPCR assays (S1 and S2) in order to reduce the risk of sample mishandling. Results: All the QF-PCR results were successful, while there was two culture failures, only one of them was repeated. No discrepancy was seen between the results of both techniques. Fifty six samples showed normal patterns, three sample showed trisomy 21, successfully detected by both techniques and one sample showed normal pattern by QF-PCR but could not be compared to the cytogenetics due to culture failure, the pregnancy outcome of this case was a normal baby. Conclusion: Our study concluded that QF-PCR is a reliable technique for prenatal diagnosis of the common chromosomal aneuploidies. It has the advantages over the cytogenetic culture of being faster with the results appearing within 24-48 hours, simpler, doesn't need a highly qualified staff, less prone to failure and more cost effective.

Keywords: QF-PCR, traditional cytogenetic fetal aneuploidies, trisomy 21, prenatal diagnosis

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625 Endeavor to Develop Immunological and Hematological Early Diagnostic Marker to Check the Conversion of Asymptomatic to Symptomatic Visceral Leishmaniasis

Authors: Roshan Kamal Topno, Maneesh Kumar, Manas Ranjan Dikhit, Krishna Pandey, Major Madhukar, Vidhya Nand Rabidas, Vahab Ali, Ganesh Chandra Sahoo, Bhawana, Devendra Prasad Yadav, Rishikesh Kumar, Pradeep Das

Abstract:

A diagnostic marker for asymptomatic subject becomes a crucial need for advocating early prophylactic majors to control protozoal infection. The main issue in epidemiological affected regions is the presence of an asymptomatic individual that might potentially convert to a symptomatic visceral leishmaniasis (VL). The epidemiological study has been conducted at highly VL endemic Moriyama village in Patna district, Bihar, India that covers total population of 1540 individuals. Here, 1104(74.02%) people had been randomly screened and only 46 (4.17%) asymptomatic individuals were found sero-positive by the rK39 test. After taking signed informed consent form, blood samples were collected from 46 asymptomatic subjects for further hematological and immunological tests. Total leukocyte count, hemoglobin (gm%), neutrophil, lymphocyte, platelet count and interleukin-10 (IL-10) had been included as diagnostic markers. Interestingly only 5 (10.86%) individuals showed their asymptomatic conversion into symptomatic VL patients during quarterly surveillance. In overall analysis only two markers are suggestive for disease conversion that is hemoglobin (gm%) and IL-10. In all the infected patients, both the mean decrease in hemoglobin and mean increase of IL-10 was 19.23% from its normal value. The results might suggest that hematological and immunological changes would become helpful for early diagnosis of asymptomatic to symptomatic VL conversion.

Keywords: asymptomatic, epidemiological, symptomatic visceral leishmaniasis, hemoglobin (gm%), interleukin-10, diagnosis

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624 Combined Aerobic-Resistance Exercise Training and Broccoli Supplementation on Plasma Decitin-1 and Insulin Resistance in Men with Type 2 Diabetes

Authors: Mohammad Soltani, Ayoub Saeidi, Nikoo Khosravi, Hanieh Nohbaradar, Seyedeh Parya Barzanjeh, Hassane Zouhal

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Exercise training and herbs supplement represent have role in the treatment for patients with type 2 diabetes (T2D). However, it is unclear combined effects of exercise training and herbs supplements on diabetic risk markers. This study aimed to determine the effect of 12 weeks of combined exercise and broccoli supplementation on decitin-1 and insulin resistance in men with type 2 diabetes. Forty-four type 2 diabetes men (age, 48.52 ± 4.36) were randomly allocated to training -supplement (TS, n = 11), training- placebo (TP, n = 11), supplement (S, n = 11) and control- placebo (CP, n = 11) groups. The combined exercise program included 12 weeks, three sessions per week, that each session contained 45 minutes of resistance training with intensity 60-70% of one maximal repetition and 30 minutes aerobic training (running) with intensity 60-70% of maximum heart rate. In addition supplement groups consumed 10 grams of Broccoli per day for 12 weeks. Plasma Decitin-1, HOMA-IR, Insulin, glucose and body composition were assessed before and after training. Plasma Dectin-1, HOMA-IR, glucose and BMI significantly decreased in TS, TP and S groups compared with CP group (P < .05). In addition Insulin and skeletal muscles mass showed significant increase in TS and TP groups compared with S and CP groups (P < .05). It is concluded that both combined exercise training (aerobic-resistance) or broccoli supplement can improve plasma Decitin-1 and insulin resistance in two diabetic patients however combine of exercise training and broccoli supplement have more effective on these markers.

Keywords: broccoli supplements, combined training, decitin-1, insulin resistance, type 2 diabetes

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623 The Markers -mm and dämmo in Amharic: Developmental Approach

Authors: Hayat Omar

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Languages provide speakers with a wide range of linguistic units to organize and deliver information. There are several ways to verbally express the mental representations of events. According to the linguistic tools they have acquired, speakers select the one that brings out the most communicative effect to convey their message. Our study focuses on two markers, -mm and dämmo, in Amharic (Ethiopian Semitic language). Our aim is to examine, from a developmental perspective, how they are used by speakers. We seek to distinguish the communicative and pragmatic functions indicated by means of these markers. To do so, we created a corpus of sixty narrative productions of children from 5-6, 7-8 to 10-12 years old and adult Amharic speakers. The experimental material we used to collect our data is a series of pictures without text 'Frog, Where are you?'. Although -mm and dämmo are each used in specific contexts, they are sometimes analyzed as being interchangeable. The suffix -mm is complex and multifunctional. It marks the end of the negative verbal structure, it is found in the relative structure of the imperfect, it creates new words such as adverbials or pronouns, it also serves to coordinate words, sentences and to mark the link between macro-propositions within a larger textual unit. -mm was analyzed as marker of insistence, topic shift marker, element of concatenation, contrastive focus marker, 'bisyndetic' coordinator. On the other hand, dämmo has limited function and did not attract the attention of many authors. The only approach we could find analyzes it in terms of 'monosyndetic' coordinator. The paralleling of these two elements made it possible to understand their distinctive functions and refine their description. When it comes to marking a referent, the choice of -mm or dämmo is not neutral, depending on whether the tagged argument is newly introduced, maintained, promoted or reintroduced. The presence of these morphemes explains the inter-phrastic link. The information is seized by anaphora or presupposition: -mm goes upstream while dämmo arrows downstream, the latter requires new information. The speaker uses -mm or dämmo according to what he assumes to be known to his interlocutors. The results show that -mm and dämmo, although all the speakers use them both, do not always have the same scope according to the speaker and vary according to the age. dämmo is mainly used to mark a contrastive topic to signal the concomitance of events. It is more commonly used in young children’s narratives (F(3,56) = 3,82, p < .01). Some values of -mm (additive) are acquired very early while others are rather late and increase with age (F(3,56) = 3,2, p < .03). The difficulty is due not only because of its synthetic structure but primarily because it is multi-purpose and requires a memory work. It highlights the constituent on which it operates to clarify how the message should be interpreted.

Keywords: acquisition, cohesion, connection, contrastive topic, contrastive focus, discourse marker, pragmatics

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622 Clinical and Chemokine Profile in Leprosy Patients During Multidrug Therapy (MDT) and Their Healthy Contacts: A Randomized Control Trial

Authors: Rohit Kothari

Abstract:

Background: Leprosyis a chronic granulomatous diseasecaused by Mycobacterium leprae (M. Lepra). Reactions may interrupt its usual chronic course.Type-1 (T1R)and type-2 lepra reaction(T2R) are acute events and signifytype-IV and type-III hypersensitivity responses, respectively. Various chemokines like CCL3, 5, 11, and CCL24 may be increased during the course of leprosy or during reactions and may serve as markers of early diagnosis, response to therapy, and prognosis. Objective: To find correlation of CCL3, 5, 11, and CCL24 in leprosy patients on multidrug therapy and their family contacts after ruling out active disease during leprosy treatment and during periods of lepra reactions. Methodology: This randomized control trial was conducted in 50 clinico-histopathologically diagnosed cases of leprosy in a tertiary care hospital in Bengaluru, India. 50 of their family contacts were adequately examined and investigated should the need be to rule out active disease. The two study-groups comprised of leprosy cases, and the age, sex, and area of residence matched healthy contactswho were given single-dose rifampicin prophylaxis, respectively. Blood samples were taken at baseline, six months, and after one yearin both the groups (on completion of MDT in leprosy cases)and also during periods of reaction if occurred in leprosy cases. Results: Our study found that at baseline, CCL5, 11, and 24 were higher in leprosy cases as compared to the healthy contacts, and the difference was statistically significant.CCL3 was also found to be higherat baseline in leprosy cases, however, the difference was not statistically significant. At six months and one year, the levels of CCL 5, 11, and 24 reduced, and the difference was statistically significant in leprosy cases, whereas it remained almost static in all the healthy contacts. Twenty patients of leprosy developed lepra reaction during the course of one year, and during reaction, the increase in CCL11 and 24 was statistically significant from baseline, whereas CCL3 and 5 did not rise significantly. One of the healthy contacts developed signs of leprosy in the form of hypopigmented numb patch and was clinico-histopathologically, and CCL11 and 24 were found to be higher with a statistically significant difference from the baseline values. Conclusion: CCL5, 11, and 24 are sensitive markers of diagnosing leprosy, response to MDT, and prognosis and are not increased in healthy contacts. CCL11 and 24 are sensitive markers of lepra reactions and may serve as one of the early diagnostic modalities for identifying lepra reaction and also leprosy in healthy contacts. To the best of our knowledge, this is the first study to evaluate these biomarkers in leprosy cases and their healthy contacts with a follow-up of upto one year with one of them developing the disease, and the same was confirmed based on these biomarkers as well.

Keywords: chemokine profile, healthy contacts, leprosy, lepra reactions

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621 Evaluation of Adaptive Fitness of Indian Teak (Tectona grandis L. F.) Metapopulation through Inter Simple Sequence Repeat Markers

Authors: Vivek Vaishnav, Shamim Akhtar Ansari

Abstract:

Teak (Tectona grandis L.f.) belonging to plant family Lamiaceae and the most commercialized timber species is endemic to South-Asia. The adaptive fitness of the species metapopulation was evaluated through its genetic differentiation and assessing the influence of geo-climatic conditions. 290 genotypes were sampled from 29 locations of its natural distribution and the genetic data was incorporated with geo-climatic parameters. Through Bayesian approach based analysis of 43 highly polymorphic ISSR markers, six homogeneous clusters (0.8% genetic variability) were identified. The six clusters were found with the various regimes of the temperature range, i.e., I - 9.10±1.35⁰C, II -6.35±0.21⁰C, III -12.21±0.43⁰C, IV - 10.8±1.06⁰C, V - 11.67±3.04⁰C, and VI - 12.35±0.21⁰C. The population had a very high percentage of LD (21.48%) among the amplified loci possibly due to experiencing restricted gene flow as well as co-adaptation and association of distant/diverse loci/alleles as a result of the stabilized climatic conditions and countless cycles of historical recombination events on a large geological timescale. The same possibly accounts for the narrow distribution of teak as a climax species in the tropical deciduous forests of the country. The regions of strong LD in teak genome significantly associated with climatic parameters also reflect that the species is tolerant to the wide regimes of the temperature range and may possibly withstand global warming and climate change in the coming millennium.

Keywords: Bayesian analysis, inter simple sequence repeat, linkage disequilibrium, marker-geoclimatic association

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620 Neuroimaging Markers for Screening Former NFL Players at Risk for Developing Alzheimer's Disease / Dementia Later in Life

Authors: Vijaykumar M. Baragi, Ramtilak Gattu, Gabriela Trifan, John L. Woodard, K. Meyers, Tim S. Halstead, Eric Hipple, Ewart Mark Haacke, Randall R. Benson

Abstract:

NFL players, by virtue of their exposure to repetitive head injury, are at least twice as likely to develop Alzheimer's disease (AD) and dementia as the general population. Early recognition and intervention prior to onset of clinical symptoms could potentially avert/delay the long-term consequences of these diseases. Since AD is thought to have a long preclinical incubation period, the aim of the current research was to determine whether former NFL players, referred to a depression center, showed evidence of incipient dementia in their structural imaging prior to diagnosis of dementia. Thus, to identify neuroimaging markers of AD, against which former NFL players would be compared, we conducted a comprehensive volumetric analysis using a cohort of early stage AD patients (ADNI) to produce a set of brain regions demonstrating sensitivity to early AD pathology (i.e., the “AD fingerprint”). A cohort of 46 former NFL players’ brain MRIs were then interrogated using the AD fingerprint. Brain scans were done using a T1-weighted MPRAGE sequence. The Free Surfer image analysis suite (version 6.0) was used to obtain the volumetric and cortical thickness data. A total of 55 brain regions demonstrated significant atrophy or ex vacuo dilatation bilaterally in AD patients vs. healthy controls. Of the 46 former NFL players, 19 (41%) demonstrated a greater than expected number of atrophied/dilated AD regions when compared with age-matched controls, presumably reflecting AD pathology.

Keywords: alzheimers, neuroimaging biomarkers, traumatic brain injury, free surfer, ADNI

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619 Modulation of Tamoxifen-Induced Cytotoxicity in Breast Cancer Cell Lines by 3-Bromopyruvate

Authors: Yasmin M. Attia, Hanan S. El-Abhar, Mahmoud M. Al Marzabani, Samia A. Shouman

Abstract:

Background: Tamoxifen (TAM) is the most commonly used hormone therapy for the treatment of early and metastatic breast cancer. Although it significantly decreases the tumor recurrence rate and provides an overall benefit, as much as 20–30% of women still relapse during or after long-term therapy. 3-Bromopyruvate (3-BP) is a promising agent with impressive antitumor effects in several models of animal tumors and cell lines. Aim: This study was designed to investigate the combined effect of (TAM) and (3-BP) in breast cancer cells and to explore their molecular interaction via assessment of apoptotic, angiogenic, and metastatic markers. Methods: In vitro cytotoxicity study was carried out for both compounds to determine the combination regimen producing a synergistic effect and mechanistic pathways were studied using RT-PCR and western techniques. Moreover, the anti-oncolytic and anti-angiogenic potentials were assessed in mice bearing solid Ehrlich carcinoma (SEC). Results: The combined treatment significantly increased the expressions and protein levels of caspase 7, 9, and 3 and decreased of angiogenic markers VEGF, HIF-1α, and HK2 compared to cells treated with either drug individually. However, there were no significant changes in MMP-2 and MMP-9 protein levels. Interestingly, the in vivo results supported the in vitro findings; there was a decrease in the tumor volume and VEFG using immunohistochemistry in the combination-treated groups compared to either TAM or 3-BP treated one. Conclusion: 3-BP synergizes the cytotoxic effect of TAM by increasing apoptosis and decreasing angiogenesis which makes this combination a promising regimen to be applied clinically.

Keywords: tamoxifen, 3-bromopyruvate, breast cancer, cytotoxicity, angiogenesis

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618 The Role of Leukocyte-Derived IL-10 on Postoperative ileus and Intestinal Macrophage Differentiation in Mice

Authors: Kathy Stein, Mariola Lysson, Anja Schmidt, Beatrix Schumak, Sabine Specht, Hicham Bouabe, Jürgen Heesemann, Axel Roers, Joerg C. Kalff, Sven Wehner

Abstract:

Objective: Postoperative ileus (POI) is a common complication of abdominal surgery. Monocyte infiltration is a hallmark of POI. The polarization of macrophages/monocytes in this process is not well understood. We aimed to investigate if and how M2 macrophage/monocyte differentiation is involved in POI pathogenesis. Design: POI was induced by intestinal manipulation (IM). C57Bl/6, CCR2-/-, IL-10 reporter (ITIB), IL-10-/- and LysMcre/IL-10fl/fl mice underwent IM. At various points in time leukocyte influx, gene and protein expression of cytokines, chemokines and M2 differentiation markers and intestinal motility were analyzed. Results: IM induced the postoperative expression of the M2 markers Arginase-1 and YM-1, predominantly in F4/80+Ly6C+ monocytes. Gene expression analyses indicated an IL-10-dependent, IL-4-independent M2 polarization of these monocytes. IL-10 dependency of M2 differentiation was confirmed in IL-10 deficient mice. Leukocytes, in the order of infiltrating monocytes, neutrophils, and resident macrophages were the main IL-10 producers during POI. IL-10 producing monocytes as well as M2 marker expression were almost absent in CCR2-deficient mice. However, postoperative IL-10 expression was not altered in CCR2-/- mice. The loss of M2 polarized monocytes neither protected CCR2-/- mice from nor affected resolution of POI. In contrast, IL-10 deficiency reduced postoperative neutrophil numbers and ameliorated POI. IL-10Ra expression was strongly induced in neutrophils but not in monocytes. Conclusion: We conclude that IL-10 counteracts POI resolution by activating IL-10Ra-expressing neutrophils in the late phase of disease while IL-10-dependent M2 differentiation is not pivotal to POI manifestation and resolution.

Keywords: interleukin-10, macrophages, neutrophils, postoperative ileus

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617 Shielding Engineered Islets with Mesenchymal Stem Cells Enhance Survival under Hypoxia by Inhibiting p38 MAPK

Authors: Bhawna Chandravanshi, Ramesh Bhonde

Abstract:

In the present study, we focused on the improvisation of islet survival in hypoxia. The Islet-like cell aggregates (ICAs) derived from Wharton's jelly mesenchymal stem cells (WJ-MSC) were cultured with and without WJ-MSC for 48h in hypoxia and normoxia and tested for their direct trophic effect on β cell survival. The WJ MSCs themselves secreted insulin upon glucose challenge and expressed the pancreatic markers at both transcription and translational level (C-peptide, Insulin, Glucagon and Glut 2). Direct contact of MSCs with ICAs facilitate the highest viability under hypoxia as evidenced by fluorescein diacetate/propidium iodide and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cytokine analysis of the co-cultured ICAs revealed amplification of anti-inflammatory cytokine-like TGFβ and TNFα accompanied by depletion of pro-inflammatory cytokines. The increment in VEGF and PDGFa was also seen showing their ability to vascularize upon transplantation. This was further accompanied by reduction in total reactive oxygen species, nitric oxide, and super oxide ions and down-regulation of Caspase3, Caspase8, p53 and up regulation of Bcl2 confirming prevention of apoptosis in ICAs. There was a significant reduction in the expression of p38 protein in the presence of MSCs making the ICAs responsive to glucose. Taken together our data demonstrate for the first time that the WJ-MSC expressed pancreatic markers and their supplementation protected engineered islets against hypoxia, oxidative stress, and inflammatory cytokines by inhibiting p38 MAPK protein.

Keywords: hypoxia, islet-like cell aggregates, inflammatory cytokines, oxidative stress

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616 Impact of Cd and Pb Impregnation on the Health of an Adult Population Neighbouring a Landfill

Authors: M. Cabral, A. Verdin, G. Garçon, A. Touré, C. Diop, M. Fall, S. Bouhsina, D. Dewaele, F.Cazier, A. Tall Dia, P. Shirali, A. Diouf

Abstract:

This case-control study dealt with the health adverse effects within the population neighboring the Mbeubeuss waste dump, which is located near the district of Malika (Diamalaye II) in Dakar (Senegal). All the household and industrial waste arising from Dakar are stored in this open landfill without being covered and are therefore possible sources of Pb and Cd contaminated air emissions and lixiviates. The objective of this study is part of improving the health of the population neighboring Mbeubeuss by determining Pb and Cd concentrations both in environment and humans, and studying possible renal function alterations within the adults. Soil and air samples were collected in the control site (Darou Salam) and the waste dump neighboring site (Diamalaye II). Control and exposed adults were recruited as living in Darou Salam (n = 52) and in Diamalaye II (n = 77). Pb and Cd concentrations in soil, air and biological samples were determined. Moreover, we were interested in analyzing some impregnation (zinc protoporphyrin, d-aminolevulinic acid dehydratase) and oxidative stress biomarkers (malonedialdehyde, gluthatione status), in addition to several nephrotoxicity parameters (creatinuria, proteinuria, lactate dehydrogenase, CC16 protein, glutathione S-transferase-alpha and retinol binding protein) in blood and/or urine. The results showed the significant Pb and Cd contamination of the soil and air samples derived from the landfill, and therefore of the neighboring population of adults. This critical exposure to environmental Pb and Cd had some harmful consequences for their health, as shown by the reported oxidative stress and nephrotoxicity signs.

Keywords: Pb and Cd environmental exposure, impregnation markers, landfill, nephrotoxicity markers

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615 Effect of Omega-3 Supplementation on Stunted Egyptian Children at Risk of Environmental Enteric Dysfunction: An Interventional Study

Authors: Ghada M. El-Kassas, Maged A. El Wakeel, Salwa R. El-Zayat

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Background: Environmental enteric dysfunction (EED) is asymptomatic villous atrophy of the small bowel that is prevalent in the developing world and is associated with altered intestinal function and integrity. Evidence has suggested that supplementary omega-3 might ameliorate this damage by reducing gastrointestinal inflammation and may also benefit cognitive development. Objective: We tested whether omega-3 supplementation improves intestinal integrity, growth, and cognitive function in stunted children predicted to have EED. Methodology: 100 Egyptian stunted children aged 1-5 years and 100 age and gender-matched normal children as controls. At the primary phase of the study, we assessed anthropometric measures and fecal markers such as myeloperoxidase (MPO), neopterin (NEO), and alpha-1-anti-trypsin (AAT) (as predictors of EED). Cognitive development was assessed (Bayley or Wechsler scores). Oral n-3 (omega-3) LC-PUFA at a dosage of 500 mg/d was supplemented to all cases and followed up for 6 months after which the 2ry phase of the study included the previous clinical, laboratory and cognitive assessment. Results: Fecal inflammatory markers were significantly higher in cases compared to controls. (MPO), (NEO) and (AAT) showed a significant decline in cases at the end of the 2ry phase (P < 0.001 for all). Omega-3 supplementation resulted also in a significant increase in mid-upper arm circumference (MUAC) (P < 0.01), weight for age z-score, and skinfold thicknesses (P< 0.05 for both). Cases showed significant improvement of cognitive function at phase 2 of the study. Conclusions: Omega-3 supplementation successfully improved intestinal inflammatory state related to EED. Also, some improvement of anthropometric and cognitive parameters showed obvious improvement with omega-3 supplementation.

Keywords: cognitive functions, EED, omega-3, stunting

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614 Multimodal Integration of EEG, fMRI and Positron Emission Tomography Data Using Principal Component Analysis for Prognosis in Coma Patients

Authors: Denis Jordan, Daniel Golkowski, Mathias Lukas, Katharina Merz, Caroline Mlynarcik, Max Maurer, Valentin Riedl, Stefan Foerster, Eberhard F. Kochs, Andreas Bender, Ruediger Ilg

Abstract:

Introduction: So far, clinical assessments that rely on behavioral responses to differentiate coma states or even predict outcome in coma patients are unreliable, e.g. because of some patients’ motor disabilities. The present study was aimed to provide prognosis in coma patients using markers from electroencephalogram (EEG), blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) and [18F]-fluorodeoxyglucose (FDG) positron emission tomography (PET). Unsuperwised principal component analysis (PCA) was used for multimodal integration of markers. Methods: Approved by the local ethics committee of the Technical University of Munich (Germany) 20 patients (aged 18-89) with severe brain damage were acquired through intensive care units at the Klinikum rechts der Isar in Munich and at the Therapiezentrum Burgau (Germany). At the day of EEG/fMRI/PET measurement (date I) patients (<3.5 month in coma) were grouped in the minimal conscious state (MCS) or vegetative state (VS) on the basis of their clinical presentation (coma recovery scale-revised, CRS-R). Follow-up assessment (date II) was also based on CRS-R in a period of 8 to 24 month after date I. At date I, 63 channel EEG (Brain Products, Gilching, Germany) was recorded outside the scanner, and subsequently simultaneous FDG-PET/fMRI was acquired on an integrated Siemens Biograph mMR 3T scanner (Siemens Healthineers, Erlangen Germany). Power spectral densities, permutation entropy (PE) and symbolic transfer entropy (STE) were calculated in/between frontal, temporal, parietal and occipital EEG channels. PE and STE are based on symbolic time series analysis and were already introduced as robust markers separating wakefulness from unconsciousness in EEG during general anesthesia. While PE quantifies the regularity structure of the neighboring order of signal values (a surrogate of cortical information processing), STE reflects information transfer between two signals (a surrogate of directed connectivity in cortical networks). fMRI was carried out using SPM12 (Wellcome Trust Center for Neuroimaging, University of London, UK). Functional images were realigned, segmented, normalized and smoothed. PET was acquired for 45 minutes in list-mode. For absolute quantification of brain’s glucose consumption rate in FDG-PET, kinetic modelling was performed with Patlak’s plot method. BOLD signal intensity in fMRI and glucose uptake in PET was calculated in 8 distinct cortical areas. PCA was performed over all markers from EEG/fMRI/PET. Prognosis (persistent VS and deceased patients vs. recovery to MCS/awake from date I to date II) was evaluated using the area under the curve (AUC) including bootstrap confidence intervals (CI, *: p<0.05). Results: Prognosis was reliably indicated by the first component of PCA (AUC=0.99*, CI=0.92-1.00) showing a higher AUC when compared to the best single markers (EEG: AUC<0.96*, fMRI: AUC<0.86*, PET: AUC<0.60). CRS-R did not show prediction (AUC=0.51, CI=0.29-0.78). Conclusion: In a multimodal analysis of EEG/fMRI/PET in coma patients, PCA lead to a reliable prognosis. The impact of this result is evident, as clinical estimates of prognosis are inapt at time and could be supported by quantitative biomarkers from EEG, fMRI and PET. Due to the small sample size, further investigations are required, in particular allowing superwised learning instead of the basic approach of unsuperwised PCA.

Keywords: coma states and prognosis, electroencephalogram, entropy, functional magnetic resonance imaging, machine learning, positron emission tomography, principal component analysis

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613 Neuroprotective Effects of Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg, a Peptide Isolated from Lupinus angustifolius L. Protein Hydrolysate

Authors: Maria Del Carmen Millan-Linares, Ana Lemus Conejo, Rocio Toscano, Alvaro Villanueva, Francisco Millan, Justo Pedroche, Sergio Montserrat-De La Paz

Abstract:

GPETAFLR (Glycine-Proline-Glutamine-Threonine-Alanine-Phenylalanine-Leucine-Arginine) is a peptide isolated from Lupinus angustifolius L. protein hydrolysate (LPH). Herein, the effect of this peptide was investigated in two different models of neuroinflammation: in the immortalized murine microglia cell line BV-2 and in a high-fat-diet-induced obesity mouse model. Methods and Results: Effects of GPETAFLR on neuroinflammation were evaluated by RT-qPCR, flow cytometry, and ELISA techniques. In BV-2 microglial cells, Lipopolysaccharides (LPS) enhanced the release of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) whereas GPETAFLR decreased pro-inflammatory cytokine levels and increased the release of the anti-inflammatory cytokine IL-10 in BV2 microglial cells. M1 (CCR7 and iNOS) and M2 (Arg-1 and Ym-1) polarization markers results showed how the GPETAFLR octapeptide was able to decrease M1 polarization marker expression and increase the M2 polarization marker expression compared to LPS. Animal model results indicate that GPETAFLR has an immunomodulatory capacity, both decreasing pro-inflammatory cytokine IL-6 and increasing the anti-inflammatory cytokine IL-10 in brain tissue. Polarization markers in the brain tissue were also modulated by GPETAFLR that decreased the pro-inflammatory expression (M1) and increased the anti-inflammatory expression (M2). Conclusion: Our results suggest that GPETAFLR isolated from LPH has significant potential for management of neuroinflammatory conditions and offer benefits derived from the consumption of Lupinus angustifolius L. in the prevention of neuroinflammatory-related diseases.

Keywords: GPETAFLR peptide, BV-2 cell line, neuroinflammation, cytokines, high-fat-diet

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612 Use of RAPD and ISSR Markers in Detection of Genetic Variation among Colletotrichum falcatum Went Isolates from South Gujarat India

Authors: Prittesh Patel, Rushabh Shah, Krishnamurthy Ramar, Vakulbhushan Bhaskar

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The present research work aims at finding genetic differences in the genomes of sugarcane red rot isolates Colletotrichum falcatum Went using Random Amplified Polymorphic DNA (RAPD) and interspersed simple sequence repeat (ISSR) molecular markers. Ten isolates of C. falcatum isolated from different red rot infected sugarcane cultivars stalk were used in present study. The amplified bands were scored across the lanes obtained in 15 RAPD primes and 21 ISSR primes successfully. The data were analysed using NTSYSpc 2.2 software. The results showed 80.6% and 68.07% polymorphism in RPAD and ISSR analysis respectively. Based on the RAPD analysis, ten genotypes were grouped into two major clusters at a cut-off value of 0.75. Geographically distant C. falcatum isolate cfGAN from south Gujarat had a level of similarity with Coimbatore isolate cf8436 presented on separate clade of bootstrapped dendrograms. First and second cluster consisted of five and three isolates respectively, indicating the close relation among them. The 21 ISSR primers produced 119 distinct and scorable loci in that 38 were monomorphic. The number of scorable loci for each primer varied from 2 (ISSR822) to 8 (ISSR807, ISSR823 and ISSR15) with an average of 5.66 loci per primer. Primer ISSR835 amplified the highest number of bands (57), while only 16 bands were obtained by primers ISSR822. Four primers namely ISSR830, ISSR845, ISSR4 and ISSR15 showed the highest value of percentage of polymorphism (100%). The results indicated that both of the marker systems RAPD and ISSR, individually can be effectively used in determination of genetic relationship among C falcatum accessions collected from different parts of south Gujarat.

Keywords: Colletotrichum falcatum, ISSR, RAPD, Red Rot

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611 Assessment of Genetic Diversity and Population Structure of Goldstripe Sardinella, Sardinella gibbosa in the Transboundary Area of Kenya and Tanzania Using mtDNA and msDNA Markers

Authors: Sammy Kibor, Filip Huyghe, Marc Kochzius, James Kairo

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Goldstripe Sardinella, Sardinella gibbosa, (Bleeker, 1849) is a commercially and ecologically important small pelagic fish common in the Western Indian Ocean region. The present study aimed to assess genetic diversity and population structure of the species in the Kenya-Tanzania transboundary area using mtDNA and msDNA markers. Some 630 bp sequence in the mitochondrial DNA (mtDNA) Cytochrome C Oxidase I (COI) and five polymorphic microsatellite DNA loci were analyzed. Fin clips of 309 individuals from eight locations within the transboundary area were collected between July and December 2018. The S. gibbosa individuals from the different locations were distinguishable from one another based on the mtDNA variation, as demonstrated with a neighbor-joining tree and minimum spanning network analysis. None of the identified 22 haplotypes were shared between Kenya and Tanzania. Gene diversity per locus was relatively high (0.271-0.751), highest Fis was 0.391. The structure analysis, discriminant analysis of Principal component (DAPC) and the pair-wise (FST = 0.136 P < 0.001) values after Bonferroni correction using five microsatellite loci provided clear inference on genetic differentiation and thus evidence of population structure of S. gibbosa along the Kenya-Tanzania coast. This study shows a high level of genetic diversity and the presence of population structure (Φst =0.078 P < 0.001) resulting to the existence of four populations giving a clear indication of minimum gene flow among the population. This information has application in the designing of marine protected areas, an important tool for marine conservation.

Keywords: marine connectivity, microsatellites, population genetics, transboundary

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610 Prognostic Value of Tumor Markers in Younger Patients with Breast Cancer

Authors: Lola T. Alimkhodjaeva, Lola T. Zakirova, Soniya S. Ziyavidenova

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Background: Breast cancer occupies the first place among the cancer in women in the world. It is urgent today to study the role of molecular markers which are capable of predicting the dynamics and outcome of the disease. The aim of this study is to define the prognostic value of the content of estrogen receptor (ER), progesterone receptor (PgR), and amplification of HER-2 / neu oncoprotein by studying 3 and 5-year overall and relapse-free survival in 470 patients with primary operable and 280 patients with locally–advanced breast cancer. Materials and methods: Study results of 3 and 5-year overall and relapse-free survival, depending on the content of RE, PgR in primary operable patients showed that ER positive (+) and PgR (+) survival was 100 (96.2%) and 97.3 (94.6%), for ER negative (-) and PgR (-) - 69.2 (60.3%) and 65.4 (57.7%), for ER positive (+) and negative PgR (-) 87.4 (80.1%) and 81.5 (79.3%), for ER negative (-) and positive PgR (+) - 97.4 (93.4%) and 90.4 (88.5%), respectively. Survival results depended also on the level of HER-2 / neu expression. In patients with HER-2 / neu negative the survival rates were as follows: 98.6 (94.7%) and 96.2 (92.3%). In group of patients with the level of HER-2 / neu (2+) expression these figures were: 45.3 (44.3%) and 45.1 (40.2%), and in group of patients with the level of HER-2 / neu (3+) expression - 41.2 (33.1%) and 34.3 (29.4%). The combination of ER negative (-), PgR (-), HER-2 / neu (-) they were 27.2 (25.4%) and 19.5 (15.3%), respectively. In patients with locally-advanced breast cancer the results of 3 and 5-year OS and RFS for ER (+) and PgR (+) were 76.3 (69.3%) and 62.2 (61.4%), for ER (-) and RP (-) 29.1 (23.7%) and 18.3 (12.6%), for ER (+) and PgR (-) 61.2 (47.2%) and 39.4 (25.6%), for ER (-) and PgR (+) 54.3 (43.1%) and 41.3 (18.3%), respectively. The level of HER-2 / neu expression also affected the survival results. Therefore, in HER-2/ neu negative patients the survival rate was 74.1 (67.6%) and 65.1 (57.3%), with the level of expression (2+) 20.4 (14.2%) and 8.6 (6.4%), with the level of expression (3+) 6.2 (3.1%) and 1.2 (1.5%), respectively. The combination for ER, PgR, HER-2 / neu negative was 22.1 (14.3%) and 8.4 (1.2%). Conclusion: Thus, the presence of steroid hormone receptors in breast tumor tissues at primary operable and locally- advanced process as the lack of HER-2/neu oncoprotein correlates with the highest rates of 3- and 5-year overall and relapse-free survival. The absence of steroid hormone receptors as well as of HER-2/neu overexpression in malignant breast tissues significantly degrades the 3- and 5-year overall and relapse-free survival. Tumors with ER, PgR and HER-2/neu negative have the most unfavorable prognostics.

Keywords: breast cancer, estrogen receptor, oncoprotein, progesterone receptor

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609 Protective Effect of Celosia Argentea Leaf Extract on Cadmium Induced Toxicity and Oxidative Stress in Rats

Authors: Sulyman Abdulhakeem Olarewaju, S. O. Malomo, M. T. Yakubu, J. O. Akolade

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The ameliorative effect of Celosia argentea var. cristata leaf extract against cadmium (Cd) induced oxidative stress and toxicity in selected tissues of rats was investigated. Toxicity coupled with oxidative stress was induced in rats by oral administration of Cd (8 mg/kg b. wt). Preliminary quantitative phytochemical and in vitro antioxidant analyses showed that the methanolic extract of C. argentea leaves was constituted by polyphenols (5.72%), saponins (3.20%), tannins (0.65%) and cadenolides (0.006%). IC50 of 9800, 7406, and 45.04 μg/ml were recorded for inhibition of linoleic acid oxidation, 2, 2-diphenyl-1-picrylhydrazyl and hydrogen peroxide radicals respectively. Simultaneous administration of C. argentea leaf extract with Cd significantly attenuated Cd-induced elevation of serum enzyme markers such as aspartate and alanine transaminase, alkaline and acid phosphatase as well as γ-glutaryltransferase in a dose-dependent fashion, while their reduced level in the liver were significantly increased. Higher levels of enzymatic antioxidants; superoxide dismutase and catalase activities were observed in the liver, brain, kidney and testes of the Cd-induced rats treated with C. argentea extract, while lipid peroxidation expressed in malondialdehyde concentrations were lower when compared to values in rats administered Cd only. Other Cd-induced toxicity and stress markers in the serum viz. reduced uric acid and albumin levels as well as elevated total and unconjugated bilirubin were attenuated by the extract and their values compared favorably with those animals co-administered cadmium with ascorbic acid. Data from the study showed that oral administration of extract from the leaf C. argentea may ameliorate Cd-induced oxidative stress and toxicity in rats.

Keywords: toxicity, cadmium, celosia, antioxidants, oxidative stress

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