Search results for: platelet mitochondrial metabolism
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 772

Search results for: platelet mitochondrial metabolism

412 Cereal Bioproducts Conversion to Higher Value Feed by Using Pediococcus Strains Isolated from Spontaneous Fermented Cereal, and Its Influence on Milk Production of Dairy Cattle

Authors: Vita Krungleviciute, Rasa Zelvyte, Ingrida Monkeviciene, Jone Kantautaite, Rolandas Stankevicius, Modestas Ruzauskas, Elena Bartkiene

Abstract:

The environmental impact of agricultural bioproducts from the processing of food crops is an increasing concern worldwide. Currently, cereal bran has been used as a low-value ingredient for both human consumption and animal feed. The most popular bioprocessing technologies for cereal bran nutritional and technological functionality increasing are enzymatic processing and fermentation, and the most popular starters in fermented feed production are lactic acid bacteria (LAB) including pediococci. However, the ruminant digestive system is unique, there are billions of microorganisms which help the cow to digest and utilize nutrients in the feed. To achieve efficient feed utilization and high milk yield, the microorganisms must have optimal conditions, and the disbalance of this system is highly undesirable. Pediococcus strains Pediococcus acidilactici BaltBio01 and Pediococcus pentosaceus BaltBio02 from spontaneous fermented rye were isolated (by rep – PCR method), identified, and characterized by their growth (by Thermo Bioscreen C automatic turbidometer), acidification rate (2 hours in 2.5 pH), gas production (Durham method), and carbohydrate metabolism (by API 50 CH test ). Antimicrobial activities of isolated pediococcus against variety of pathogenic and opportunistic bacterial strains previously isolated from diseased cattle, and their resistance to antibiotics were evaluated (EFSA-FEEDAP method). The isolated pediococcus strains were cultivated in barley/wheat bran (90 / 10, m / m) substrate, and developed supplements, with high content of valuable pediococcus, were used for Lithuanian black and white dairy cows feeding. In addition, the influence of supplements on milk production and composition was determined. Milk composition was evaluated by the LactoScope FTIR” FT1.0. 2001 (Delta Instruments, Holland). P. acidilactici BaltBio01 and P. pentosaceus BaltBio02 demonstrated versatile carbohydrate metabolism, grown at 30°C and 37°C temperatures, and acidic tolerance. Isolated pediococcus strains showed to be non resistant to antibiotics, and having antimicrobial activity against undesirable microorganisms. By barley/wheat bran utilisation using fermentation with selected pediococcus strains, it is possible to produce safer (reduced Enterobacteriaceae, total aerobic bacteria, yeast and mold count) feed stock with high content of pediococcus. Significantly higher milk yield (after 33 days) by using pediococcus supplements mix for dairy cows feeding could be obtained, while similar effect by using separate strains after 66 days of feeding could be achieved. It can be stated that barley/wheat bran could be used for higher value feed production in order to increase milk production. Therefore, further research is needed to identify what is the main mechanism of the positive action.

Keywords: barley/wheat bran, dairy cattle, fermented feed, milk, pediococcus

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411 The Evaluation of Subclinical Hypothyroidism in Children with Morbid Obesity

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Cardiovascular pathology is one of the expected consequences of excessive fat gain. The role of zinc in thyroid hormone metabolism is an important matter. The concentrations of both thyroid stimulating hormone (TSH) and zinc are subject to variation in obese individuals. Zinc exhibits protective effects on cardiovascular health and is inversely correlated with cardiovascular markers in childhood obesity. The association between subclinical hypothyroidism (SCHT) and metabolic disorders is under investigation due to its clinical importance. Underactive thyroid gland causes high TSH levels. Subclinical hypothyroidism is defined as the elevated serum TSH levels in the presence of normal free thyroxin (T4) concentrations. The aim of this study was to evaluate the associations between TSH levels and zinc concentrations in morbid obese (MO) children exhibiting SCHT. The possibility of using the probable association between these parameters was also evaluated for the discrimination of metabolic syndrome positive (MetS+) and metabolic syndrome negative (MetS-) groups. Forty-two children were present in each group. Informed consent forms were obtained. Institutional Ethics Committee approved the study protocol. Tables prepared by World Health Organization were used for the definition of MO children. Children, whose age- and sex-dependent body mass index percentile values were above 99, were defined as MO. Children with at least two MetS components were included in MOMetS+ group. Elevated systolic/diastolic blood pressure values, increased fasting blood glucose, triglycerides (TRG)/decreased high density lipoprotein-cholesterol (HDL-C) concentrations in addition to central obesity were listed as MetS components. Anthropometric measures were recorded. Routine biochemical analyses were performed. Thirteen and fifteen children had SCHT in MOMetS- and MOMetS+ groups, respectively. Statistical analyses were performed. p<0.05 was accepted as statistically significant. In MOMetS- and MOMetS+ groups, TSH levels were 4.1±2.9 mU/L and 4.6±3.1 mU/L, respectively. Corresponding values for SCHT cases in these groups were 7.3±3.1 mU/L and 8.0±2.7 mU/L. Free T4 levels were within normal limits. Zinc concentrations were negatively correlated with TSH levels in both groups. The significant negative correlation calculated in MOMetS+ group (r= -0.909; p<0.001) was much stronger than that found in MOMetS- group (r= -0.706; p<0.05). This strong correlation (r= -0.909; p<0.001) calculated for cases with SCHT in MOMetS+ group was much lower (r= -0.793; p<0.001) when all MOMetS+ cases were considered. Zinc is closely related to T4 and TSH therefore, it participates in thyroid hormone metabolism. Since thyroid hormones are required for zinc absorption, hypothyroidism can lead to zinc deficiency. The presence of strong correlations between TSH and zinc in SCHT cases found in both MOMetS- and MOMetS+ groups pointed out that MO children were under the threat of cardiovascular pathologies. The detection of the much stronger correlation in MOMetS+ group in comparison with the correlation found in MOMetS- group was the indicator of greater cardiovascular risk due to the presence of MetS. In MOMetS+ group, correlation in SCHT cases found higher than correlation calculated for all cases confirmed much higher cardiovascular risk due to the contribution of SCHT.

Keywords: cardiovascular risk, children, morbid obesity, subclinical hypothyroidism, zinc

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410 Differential Diagnosis of Malaria and Dengue Fever on the Basis of Clinical Findings and Laboratory Investigations

Authors: Aman Ullah Khan, Muhammad Younus, Aqil Ijaz, Muti-Ur-Rehman Khan, Sayyed Aun Muhammad, Asif Idrees, Sanan Raza, Amar Nasir

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Dengue fever and malaria are important vector-borne diseases of public health significance affecting millions of people around the globe. Dengue fever is caused by Dengue virus while malaria is caused by plasmodium protozoan. Generally, the consequences of Malaria are less severe compared to dengue fever. This study was designed to differentiate dengue fever and malaria on the basis of clinical and laboratory findings and to compare the changes in both diseases having different causative agents transmitted by the common vector. A total of 200 patients of dengue viral infection (120 males, 80 females) were included in this prospective descriptive study. The blood samples of the individuals were first screened for malaria by blood smear examination and then the negative samples were tested by anti-dengue IgM strip. The strip positive cases were further screened by IgM capture ELISA and their complete blood count including hemoglobin estimation (Hb), total and differential leukocyte counts (TLC and DLC), erythrocyte sedimentation rate (ESR) and platelet counts were performed. On the basis of the severity of signs and symptoms, dengue virus infected patients were subdivided into dengue fever (DF) and dengue hemorrhagic fever (DHF) comprising 70 and 100 confirmed patients, respectively. On the other hand, 30 patients were found infected with Malaria while overall 120 patients showed thrombocytopenia. The patients of DHF were found to have more leucopenia, raised hemoglobin level and thrombocytopenia < 50,000/µl compared to the patients belonging to DF and malaria. On the basis of the outcomes of the study, it was concluded that patients affected by DF were at a lower risk of undergoing haematological disturbance than suffering from DHF. While, the patients infected by Malaria were found to have no significant change in their blood components.

Keywords: dengue fever, blood, serum, malaria, ELISA

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409 Morphometric Parameters and Evaluation of Persian Fallow Deer Semen in Dashenaz Refuge in Iran

Authors: Behrang Ekrami, Amin Tamadon

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Persian fallow deer (Dama dama mesopotamica) is belonging to the family Cervidae and is only found in a few protected areas in the northwest, north, and southwest of Iran. The aims of this study were analysis of inbreeding and morphometric parameters of semen in male Persian fallow deer to investigate the cause of reduced fertility of this endangered species in Dasht-e-Naz National Refuge, Sari, Iran. The Persian fallow deer semen was collected from four adult bucks randomly during the breeding and non-breeding season from five dehorned and horned deer's BY an artificial vagina. Twelve blood samples was taken from Persian fallow deer and mitochondrial DNA was extracted, amplified, extracted, sequenced, and then were considered for genetic analysis. The Persian fallow deer semen, both with normal and abnormal spermatozoa, is similar to that of domestic ruminants but very smaller and difficult to observe at the primary observation. The post-mating season collected ejaculates contained abnormal spermatozoa, debris and secretion of accessory glands in horned bucks and accessory glands secretion free of any spermatozoa in dehorned or early velvet budding bucks. Microscopic evaluation in all four bucks during the mating season showed the mean concentration of 9×106 spermatozoa/ml. The mean ±SD of age, testes length and testes width was 4.60±1.52 years, 3.58±0.32 and 1.86±0.09 cm, respectively. The results identified 1120 loci (assuming each nucleotide as locus) in which 377 were polymorphic. In conclusion, reduced fertility of male Persian fallow deer may be caused by inbreeding of the protected herd in a limited area of Dasht-e-Naz National Refuge.

Keywords: Persian fallow deer, spermatozoa, reproductive characteristics, morphometric parameters

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408 Revisiting Classic Triad of Japanese Spotted Fever: A Case Series of Forty-Three Patients

Authors: Y. Kunitani, Y. Nakashima, S. Yamauchi, Y. Ishigami, K. Naito, K. Numata, M. Mizobe, Y. Homma, J. Takahashi, T. Inoue, T. Shiga, H. Funakoshi

Abstract:

Background: Japanese Spotted Fever (JSF) is one of the Rickettsial infections, caused by Rickettsia japonica, which is transmitted by ticks. JSF is seen in limited area, such as Japan and South Korea. Its clinical triad is rash, eschar and fever. It often shows leukocytopenia, thrombopenia, elevated transaminase and high C-reactive protein (CRP). Sometimes it can be life-threatening due to disseminated intravascular coagulation or multiple organ failure. Study Aim: The aim of this study is to describe the features of JSF, as this unique infection is rapidly growing in Japan. Methods: This is a case series of JSF from 2009 to 2016, in Mie Prefectural Hospital in Japan. We collected JSF cases, which were diagnosed by polymerase chain reaction (PCR) of the skin or blood serum, or the elevation of the antibody titer of paired blood samples. Results: There were 43 JSF patients (19 male, 24 female) with a median age of 71 years [IQR:65-80]. The median body temperature was 38.1°C[IQR: 37.5-39.0]. 95% had a rash, 67% had eschar and 50% had fever. The median WBC count was 6,700 [IQR: 5,750-8,200] and leukocytopenia was observed in only 7%. The median platelet count was 14x104 [IQR10x104-17x104], thrombopenia was observed in 65%. The median aspartate transaminase (AST) was 53 IU/L [IQR: 41-93]; the median alanine aminotransferase (ALT) was 34 IU/L [IQR: 24-54]; the median CRP was 10.4 mg/dL [IQR:7.2-13.9]; the median lactate dehydrogenase (LDH) was 352IU/L [IQR:282-451]. CRP and LDH were elevated in almost all of the patients. Median length of stay in hospital was 8 days [IQR: 6-11]. All patients were treated with tetracycline and quinolone on the day of the presentation. There was no fatality from JSF. Conclusion: The patients with JSF classically presents with eschar, rash and fever. However, in this study, the half of the patients were afebrile. Although JSF is not a common infectious disease worldwide, if the patient had previously visited Japan or South Korea and presented with rash and eschar with or without fever, we should consider JSF as a potential diagnosis.

Keywords: infectious disease, Japanese spotted fever, Rickettsial disease, Rickettsia japonica

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407 Duplex Real-Time Loop-Mediated Isothermal Amplification Assay for Simultaneous Detection of Beef and Pork

Authors: Mi-Ju Kim, Hae-Yeong Kim

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Product mislabeling and adulteration have been increasing the concerns in processed meat products. Relatively inexpensive pork meat compared to meat such as beef was adulterated for economic benefit. These food fraud incidents related to pork were concerned due to economic, religious and health reasons. In this study, a rapid on-site detection method using loop-mediated isothermal amplification (LAMP) was developed for the simultaneous identification of beef and pork. Each specific LAMP primer for beef and pork was designed targeting on mitochondrial D-loop region. The LAMP assay reaction was performed at 65 ℃ for 40 min. The specificity of each primer for beef and pork was evaluated using DNAs extracted from 13 animal species including beef and pork. The sensitivity of duplex LAMP assay was examined by serial dilution of beef and pork DNAs, and reference binary mixtures. This assay was applied to processed meat products including beef and pork meat for monitoring. Each set of primers amplified only the targeted species with no cross-reactivity with animal species. The limit of detection of duplex real-time LAMP was 1 pg for each DNA of beef and pork and 1% pork in a beef-meat mixture. Commercial meat products that declared the presence of beef and/or pork meat on the label showed positive results for those species. This method was successfully applied to detect simultaneous beef and pork meats in processed meat products. The optimized duplex LAMP assay can identify simultaneously beef and pork meat within less than 40 min. A portable real-time fluorescence device used in this study is applicable for on-site detection of beef and pork in processed meat products. Thus, this developed assay was considered to be an efficient tool for monitoring meat products.

Keywords: beef, duplex real-time LAMP, meat identification, pork

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406 Genomic Adaptation to Local Climate Conditions in Native Cattle Using Whole Genome Sequencing Data

Authors: Rugang Tian

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In this study, we generated whole-genome sequence (WGS) data from110 native cattle. Together with whole-genome sequences from world-wide cattle populations, we estimated the genetic diversity and population genetic structure of different cattle populations. Our findings revealed clustering of cattle groups in line with their geographic locations. We identified noticeable genetic diversity between indigenous cattle breeds and commercial populations. Among all studied cattle groups, lower genetic diversity measures were found in commercial populations, however, high genetic diversity were detected in some local cattle, particularly in Rashoki and Mongolian breeds. Our search for potential genomic regions under selection in native cattle revealed several candidate genes related with immune response and cold shock protein on multiple chromosomes such as TRPM8, NMUR1, PRKAA2, SMTNL2 and OXR1 that are involved in energy metabolism and metabolic homeostasis.

Keywords: cattle, whole-genome, population structure, adaptation

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405 Bioinformatics Analysis of DGAT1 Gene in Domestic Ruminnants

Authors: Sirous Eydivandi

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Diacylglycerol-O-acyltransferase (DGAT1) gene encodes diacylglycerol transferase enzyme that plays an important role in glycerol lipid metabolism. DGAT1 is considered to be the key enzyme in controlling the synthesis of triglycerides in adipocytes. This enzyme catalyzes the final step of triglyceride synthesis (transform triacylglycerol (DAG) into triacylglycerol (TAG). A total of 20 DGAT1 gene sequences and corresponding amino acids belonging to 4 species include cattle, goats, sheep and yaks were analyzed, and the differentiation within and among the species was also studied. The length of the DGAT1 gene varies greatly, from 1527 to 1785 bp, due to deletion, insertion, and stop codon mutation resulting in elongation. Observed genetic diversity was higher among species than within species, and Goat had more polymorphisms than any other species. Novel amino acid variation sites were detected within several species which might be used to illustrate the functional variation. Differentiation of the DGAT1 gene was obvious among species, and the clustering result was consistent with the taxonomy in the National Center for Biotechnology Information.

Keywords: DGAT1gene, bioinformatic, ruminnants, biotechnology information

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404 Can Bone Resorption Reduce with Nanocalcium Particles in Astronauts?

Authors: Ravi Teja Mandapaka, Prasanna Kumar Kukkamalla

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Poor absorption of calcium, elevated levels in serum and loss of bone are major problems of astronauts during space travel. Supplementation of calcium could not reveal this problem. In normal condition only 33% of calcium is absorbed from dietary sources. In this paper effect of space environment on calcium metabolism was discussed. Many surprising study findings were found during literature survey. Clinical trials on ovariectomized mice showed that reduction of calcium particles to nano level make them more absorbable and bioavailable. Control of bone loss in astronauts in critical important In Fortification of milk with nana calcium particles showed reduces urinary pyridinoline, deoxypyridinoline levels. Dietary calcium and supplementation do not show much retention of calcium in zero gravity environment where absorption is limited. So, the fortification of foods with nano calcium particles seemed beneficial for astronauts during and after space travel in their speedy recovery.

Keywords: nano calcium, astronauts, fortification, supplementation

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403 Sexually Dimorphic Effects of Chronic Exercise and Myocytic Androgen Receptor Overexpression on Body Composition in Sprague dawley Rats

Authors: Sabrina Barsky, Ashley Monks

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In humans, exercise improves symptoms of various pathological states, although exercise adaptations seem to differ in response to sex. Skeletal muscle anabolism is thought to be regulated by androgen receptor (AR) through poorly specified mechanisms. Interactions of AR and exercise on muscle phenotype remain inconclusive in males, and undetermined in females. We hypothesized that sex differences in exercise adaptations are regulated by the androgenic system and the type of exercise performed. Here we examined interactions between a muscle-specific AR overexpression transgene (HSA-AR) and forced aerobic exercise paradigm on muscle and adipose exercise adaptation in male and female rats. We used dual-energy X-ray absorptiometry (DXA) to examine body composition adaptations post 9-week exercise protocol. We replicated the effects of HSA-AR on body composition, with males only having increased % lean mass and reduced % fat mass (P<0.05). Aerobic exercise improved lean body phenotype significantly, with lesser indices of total and % fat mass (P<0.01) in both sexes. Sex-specific effects of exercise included decreased total body mass (P<0.01) in males and increased lean mass % (P<0.001) in females. Surprisingly, neither AR manipulation nor exercise affected bone parameters in either sex. This varied response in total mass and lean mass according to exercise presents a sexually dimorphic response to exercise. Neither sex showed an interaction between HSA-AR and forced aerobic exercise on body composition. Future work is proposed to examine the effects of exercise type (aerobic versus resistance) and the role of gonadal androgens in sexually dimorphic exercise-mediated mitochondrial adaptations. This work implicates the development of sex-specific exercise therapies.

Keywords: androgen receptor, forced exercise, muscle physiology, sexual dimorphism

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402 Examining the Relationship between Concussion and Neurodegenerative Disorders: A Review on Amyotrophic Lateral Sclerosis and Alzheimer’s Disease

Authors: Edward Poluyi, Eghosa Morgan, Charles Poluyi, Chibuikem Ikwuegbuenyi, Grace Imaguezegie

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Background: Current epidemiological studies have examined the associations between moderate and severe traumatic brain injury (TBI) and their risks of developing neurodegenerative diseases. Concussion, also known as mild TBI (mTBI), is however quite distinct from moderate or severe TBIs. Only few studies in this burgeoning area have examined concussion—especially repetitive episodes—and neurodegenerative diseases. Thus, no definite relationship has been established between them. Objectives : This review will discuss the available literature linking concussion and amyotrophic lateral sclerosis (ALS) and Alzheimer’s disease (AD). Materials and Methods: Given the complexity of this subject, a realistic review methodology was selected which includes clarifying the scope and developing a theoretical framework, developing a search strategy, selection and appraisal, data extraction, and synthesis. A detailed literature matrix was set out in order to get relevant and recent findings on this topic. Results: Presently, there is no objective clinical test for the diagnosis of concussion because the features are less obvious on physical examination. Absence of an objective test in diagnosing concussion sometimes leads to skepticism when confirming the presence or absence of concussion. Intriguingly, several possible explanations have been proposed in the pathological mechanisms that lead to the development of some neurodegenerative disorders (such as ALS and AD) and concussion but the two major events are deposition of tau proteins (abnormal microtubule proteins) and neuroinflammation, which ranges from glutamate excitotoxicity pathways and inflammatory pathways (which leads to a rise in the metabolic demands of microglia cells and neurons), to mitochondrial function via the oxidative pathways.

Keywords: amyotrophic lateral sclerosis, Alzheimer's disease, mild traumatic brain injury, neurodegeneration

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401 Apoptosis Inducing Potential of Onosma Bracteata Wall. in Mg-63 Human Osteosarcoma Cells via cdk2/Cyclin E Pathway

Authors: Ajay Kumar, Satwinderjeet Kaur

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Onosma bracteata Wall. (Boraginaceae), is known to be a medicinal plant, useful in the treatment of body swellings, abdominal pain and urinary calculi, etc. The present study focused on the radical scavenging and cancer growth inhibitory properties of isolates from O. bracteata. Obea fraction demonstrated noticeable free radical scavenging ability along with antiproliferative activity in human osteosarcoma MG-63, human neuroblastoma IMR-32, and human lung cancer A549 cell lines using MTT assay with GI50 values of 88.56, 101.61 and 112.7 μg/ml, respectively. The scanning electron and confocal microscopy studies showed morphological alterations including nuclear condensation and formation of apoptotic bodies in osteosarcoma MG-63 cells. Obea fraction in osteosarcoma MG-63 cells augmented the reactive oxygen species (ROS) level and decreased the mitochondrial membrane potential. Flow cytometry analysis revealed the Obea treated cells to be arrested in the G0/G1 phase in a dose dependent manner supported by the observed increase in the early apoptotic cell population. Western blotting analysis showed that the expression of p-NF-kB, COX-2, p-Akt, and Bcl-xL decreased whereas, the expression of GSK-3β, p53, caspase-3 and caspase-9 proteins increased. The downregulation of Bcl-2, Cyclin E, CDK2 and mortalin gene expression and upregulation of p53 genes was unfolded in RT-qPCR studies. The presence of catechin, kaempferol, Onosmin A and epicatechin, as revealed in high-performance liquid chromatography (HPLC) studies, contributes towards the chemopreventive potential of O. bracteata which can be tapped for chemotherapeutic use.

Keywords: apoptosis, confocal microscopy, HPLC, mitochondria membrane potential, reactive oxygen species

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400 Protective Effect of Vitamin D on Cardiac Apoptosis in Obese Rats

Authors: Kadeejah Alsolami, Zainab Alrefay, Husaam Awad

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Obesity and vitamin D deficiency have both been related to cardiovascular disease. The present work aimed to investigate the possible protective effect of vitamin D on cardiac apoptosis in a rat model of dietary-induced obesity. Methods: 30 male Wistar rats included in this study. They were allocated into 4 groups: Control (n=5), animal were fed standard diet for 3 months: Control + vitamin D (VD) (n=5),animals were fed a standard diet with 400IU VD/kg for 3 months: hypercaloric diets group (n=10), animals were fed a high fat diet for 3 months: hypercaloric diet with VD group (n=10), animals were fed a high fat diet with 400IU VD/kg for 3 months. At the beginning of the experiment, the weight and length were measured to assess body mass index (BMI) and repeated every 45 days. Food intake and body weight were monitored throughout the study period. Then rats were sacrificed and heart tissues collected for Quantitative Real-time polymerase chain reaction (qRT-PCR). qRT-PCR used to detect different genetic markers of apoptosis (anti-apoptotic gene (BCL2), a pro-apoptotic gene(BAX), pro-apoptotic genes (FAS, FAS-L), tumour necrosis factor (TNF), mitogen-activated protein kinases (MAPK). Results: FAS and FAS-L gene expression were significantly upregulated in rats fed with high fat diet. And FAS-L gene expression was significantly upregulated in all groups on comparison with control. Whereas Bax gene expression was significantly downregulated in rats fed with high-fat diet supplied with vitamin D. TNF was significantly upregulated in rats fed with high-fat diet treated with vitamin D. MAPK was significantly upregulated in rats fed with high fat diet group, and in rats fed with high-fat diet supplied with vitamin D. Conclusion: The cardiac apoptotic pathways were more activated in rats fed with high-fat than lean rats. And vitamin D protect the heart from the cardiac mitochondrial-dependent apoptotic pathway.

Keywords: apoptosis, heart, obesity, Vitamin D

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399 Coumestrol Induced Apoptosis in Breast Cancer MCF-7 Cells via Redox Cycling of Copper and ROS Generation: Implications of Copper Chelation Strategy in Cancer Treatment

Authors: Atif Zafar Khan, Swarnendra Singh, Imrana Naseem

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Breast cancer is one of the most frequent malignancies in women worldwide and a leading cause of cancer-related deaths among women. Therefore, there is a need to identify new chemotherapeutic strategies for cancer treatment. Unlike normal cells, cancer cells contain elevated copper levels which play an integral role in angiogenesis. Copper is an important metal ion associated with the chromatin DNA, particularly with guanine. Thus, targeting copper via copper-specific chelators in cancer cells can serve as effective anticancer strategy. Keeping in view these facts, we evaluated the anticancer activity and copper-dependent cytotoxic effect of coumestrol (phytoestrogen in soybean products) in breast cancer MCF-7 cells. Coumestrol inhibited proliferation and induced apoptosis in MCF-7 cells, which was prevented by copper chelator neocuproine and ROS scavengers. Coumestrol treatment induced ROS generation coupled to DNA fragmentation, up-regulation of p53/p21, cell cycle arrest at G1/S phase, mitochondrial membrane depolarization and caspases 9/3 activation. All these effects were suppressed by ROS scavengers and neocuproine. These results suggest that coumestrol targets elevated copper for redox cycling to generate ROS leading to DNA fragmentation. DNA damage leads to p53 up-regulation which directs the cell cycle arrest at G1/S phase and promotes caspase-dependent apoptosis of MCF-7 cells. In conclusion, coumestrol induces pro-oxidant cell death by chelating cellular copper to produce copper-coumestrol complexes that engages in redox cycling in breast cancer cells. Thus, targeting elevated copper levels might be a potential therapeutic strategy for selective cytotoxic action against malignant cells.

Keywords: apoptosis, breast cancer, copper chelation, coumestrol, reactive oxygens species, redox cycling

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398 Yeasts Associated to Spontaneous Date Vinegar Process

Authors: F. Halladj, H. Amellal, S. Benamara

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Current consumer trends go towards natural products defined as the products obtained by a traditional manufacturing method. Vinegar is one of those products marketed; it may be industrially obtained by a submerged (fast) or traditional (slow) processes. The latter exhibited a high quality because of its complex microbiological transformations (or two-stage fermentation) by the native must flora. Moreover, although that Acetic acid bacteria have traditionally been considered to play the leading role in vinegar production, some studies have recently highlighted that also yeasts metabolism can affect traditional vinegar chemical properties in a remarkable way. Thus, the aim of this study was to monitor a traditional slow process of vinegar as applied in the south of Algeria using date with hard texture (Degla-Beida variety) to isolate and identify the involved yeasts in order to select them as starter culture. Phenotypic and molecular analysis show that the non-Saccharomyces were the main yeasts species isolated throughout the alcoholic spontaneous fermentation and they included Hanseniaspora guilliermondii and Torulaspora delbrueckii.

Keywords: date vinegar, traditional production, yeasts, Phenotypic, Algeria

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397 Nanoparticle Induced Neurotoxicity Mediated by Mitochondria

Authors: Nandini Nalika, Suhel Parvez

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Nanotechnology has emerged to play a vital role in developing all through the industrial world with an immense production of nanomaterials including nanoparticles (NPs). Many toxicological studies have confirmed that due to unique small size and physico-chemical properties of NPs (1-100nm), they can be potentially hazardous. Metallic NPs of small size have been shown to induce higher levels of cellular oxidative stress and can easily pass through the Blood Brain Barrier (BBB) and significantly accumulate in brain. With the wide applications of titanium dioxide nanoparticles (TNPs) in day-to-day life in form of cosmetics, paints, sterilisation and so on, there is growing concern regarding the deleterious effects of TNPs on central nervous system and mitochondria appear to be important cellular organelles targeted to the pro-oxidative effects of NPs and an important source that contribute significantly for the production of reactive oxygen species after some toxicity or an injury. The aim of our study was to elucidate the effect of TNPs in anatase form with different concentrations (5-50 µg/ml) following with various oxidative stress markers in isolated brain mitochondria as an in vitro model. Oxidative stress was determined by measuring the different oxidative stress markers like lipid peroxidation as well as the protein carbonyl content which was found to be significantly increased. Reduced glutathione content and major glutathione metabolizing enzymes were also modulated signifying the role of glutathione redox cycle in the pathophysiology of TNPs. The study also includes the mitochondrial enzymes (Complex 1, Complex II, complex IV, Complex V ) and the enzymes showed toxicity in a relatively short time due to the effect of TNPs. The study provide a range of concentration that were toxic to the neuronal cells and data pointing to a general toxicity in brain mitochondria by TNPs, therefore, it is in need to consider the proper utilization of NPs in the environment.

Keywords: mitochondria, nanoparticles, brain, in vitro

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396 Impact of Heavy Metal Toxicity on Metabolic Changes in the Diazotrophic Cyanobacterium Anabaena PCC 7120

Authors: Rishi Saxena

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Cyanobacteria is a photosynthetic prokaryote, and these obtain their energy through photosynthesis. In this paper, we studied the effect of iron on metabolic changes in the diazotrophic cyanobacterium Anabaena PCC 7120. Nowadays, metal contamination due to natural and anthropogenic sources is a global environment concern. Iron induced changes in growth, N2-fixation, CO2 fixation and photosynthetic activity were studied in a diazotrophic cyanobacterium Anabaena PCC 7120. Iron at 50 uM concentration supported the maximum growth, heterocyst frequency, CO2 fixation, photosystem I (PS I), photosystem II (PS II) and nitrogenase activities in the organism. Higher concentration of iron inhibited these processes. Chl a and PS II activities were more sensitive to iron than the protein and PS I activity. Here, it is also mentioned that heavy metal induced altered macromolecules metabolism and changes in the central dogma of life (DNA→ mRNA → Protein). And also recent advances have been made in understanding heavy metal-cyanobacteria interaction and their application for metal detoxification.

Keywords: cyanobacterium anabaena 7120, nitrogen fixation, photosystem I (PS I), photosystem II (PS II)

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395 Biofilm Formation Due to the Proteome Changes Of Enterococcus Faecium in Response to Sub-Mic of Gentamicin

Authors: Amin Abbasi, Mahdi Asghari Ozma

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Background and Objective:Enterococcus faecium is a normal flora of the human gastrointestinal tract that causes infection in the host body under conditions such as biofilm formation, in which the use of antibiotics causes changes in these pathogenic mechanisms. In this study, we aimed to evaluate comprehensively the changes in E.faecium when exposed to sub-MIC of the gentamicin,especiallythe biofilm formation rate. Materials and Methods: For this study, the keywords "Enterococcus faecium ", "Biofilm", and "Gentamicin" in the databases PubMed, Google Scholar, Sid, and MagIran between 2015 and 2021 were searched, and 14 articles were chosen, studied, and analyzed. Results: Gentamicin significantly had increased biofilm formation in most of the isolates in the studies. Increased expression of the genes (efaA and esp) and proteins involved in biofilm formation and decreased expression of the genes (gelE and cylA) involved in spreading and proteins involved in metabolism and cell division in E.faecium were the most significant cause of the biofilm formation, which were increased in sub-MIC gentamicin-treated situation. Conclusion: Inadequate use of gentamicin intensify biofilm formation of E.faecium, which can make the treatment of infections caused by this bacterium difficult.

Keywords: biofilm, enterococcus faecium, gentamicin, proteome

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394 Effect of Age and Physiological Status on Some Serum Energy Metabolites and Progesterone in Ouled Djellal Breed Ewes in Algeria

Authors: B. Safsaf, M. Tlidjane, B. Mamache, M. A. Dehimi, H. Boukrous, Aly A. Hassan

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The aim of this study is to determine the effect of age and physiological status on progesterone and energy metabolism of Ouled Djellal (O.D) breed ewes. 40 healthy ewes were divided into two groups, primiparous and multiparous, with 20 ewes in each group. The body weights (BW) (kg) were 46.6 ± 4.20 and 59.2 ± 3.02, and consuming less 25 to 30% of their basal energetic requirements. The values of serum glucose, triglycerides and cholesterol were lower in pregnant than in non-pregnant ewes. The high to very high significant differences were found during the 15th week of pregnancy for glycaemia and triglyceridemia respectively. Concerning serum progesterone, a very highly significant difference (p < 0.001) was noted in the pregnant group, and the values were higher in MP than in PP. After lambing, the triglyceridemia values were slightly lower in primiparous than in multiparous pregnant ewes. In order to prevent imbalance during critical periods of reproduction, we can use the serum metabolic profile.

Keywords: age, energy metabolites, ouled djellal breed ewes, physiologic status, progesterone

Procedia PDF Downloads 557
393 Unsupervised Classification of DNA Barcodes Species Using Multi-Library Wavelet Networks

Authors: Abdesselem Dakhli, Wajdi Bellil, Chokri Ben Amar

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DNA Barcode, a short mitochondrial DNA fragment, made up of three subunits; a phosphate group, sugar and nucleic bases (A, T, C, and G). They provide good sources of information needed to classify living species. Such intuition has been confirmed by many experimental results. Species classification with DNA Barcode sequences has been studied by several researchers. The classification problem assigns unknown species to known ones by analyzing their Barcode. This task has to be supported with reliable methods and algorithms. To analyze species regions or entire genomes, it becomes necessary to use similarity sequence methods. A large set of sequences can be simultaneously compared using Multiple Sequence Alignment which is known to be NP-complete. To make this type of analysis feasible, heuristics, like progressive alignment, have been developed. Another tool for similarity search against a database of sequences is BLAST, which outputs shorter regions of high similarity between a query sequence and matched sequences in the database. However, all these methods are still computationally very expensive and require significant computational infrastructure. Our goal is to build predictive models that are highly accurate and interpretable. This method permits to avoid the complex problem of form and structure in different classes of organisms. On empirical data and their classification performances are compared with other methods. Our system consists of three phases. The first is called transformation, which is composed of three steps; Electron-Ion Interaction Pseudopotential (EIIP) for the codification of DNA Barcodes, Fourier Transform and Power Spectrum Signal Processing. The second is called approximation, which is empowered by the use of Multi Llibrary Wavelet Neural Networks (MLWNN).The third is called the classification of DNA Barcodes, which is realized by applying the algorithm of hierarchical classification.

Keywords: DNA barcode, electron-ion interaction pseudopotential, Multi Library Wavelet Neural Networks (MLWNN)

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392 DNA Methylation 6mA and Histone Methylation Involved in Multi-/Trans-Generational Reproductive Effects in Caenorhabditis elegans Induced by Atrazine

Authors: Jiechen Yin, Xiang Hong, Ran Liu

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Atrazine (ATR), a widely used triazine herbicide, is an environmental endocrine disruptor that can cause health problems. However, whether there are multi/trans-generational reproductive impacts of ATR have not been studied to our best knowledge. Therefore, in this study, Caenorhabditis elegans was used as a preferable model organism to identify the multi/trans-generational reproductive toxicity of ATR. L1 larvae were exposed to different concentrations (0.0004–40 mg/L) of ATR for 48 h. Successive generations (F1 to F5) were fed without ATR and consecutive exposure. The results showed that ATR exposure during P0 decreased fecundity, including a reduction in fertilized eggs, oocytes, and ovulation rate, delayed gonadal development, and decreased the relative area of the gonad arm and germ cell number. Furthermore, continuous ATR exposure (P0–F5) causes a significant increase in reproductive toxicity in subsequent generations, although no significant toxicity occurred in the P0 generation after exposure to environmental-related concentrations, suggesting that ATR exposure might have cumulative effects. Likewise, parental exposure to ATR caused transgenerational toxicity impairments. Interestingly, reproductive toxicity not development toxicity was transmitted to several generations (F1–F4), and the F2 generation showed the most notable changes. QRT-PCR results showed that genes related to DNA methylation 6mA (damt-1, nmad-1) and histone H3 methylation (mes-4, met-2, set-25, set-2, and utx-1) can also be passed on to offspring. The function of H3K4 and H3K9 methylation were explored by using loss-of-function mutants for set-2, set-25, and met-2. Transmissible reproductive toxicity was absent in met-2(n4256), set-2(ok952), and set-25(n5021) mutants, which suggests that the histone methyltransferases H3K4 and H3K9 activity are indispensable for the transgenerational effect of ATR. Finally, the downstream genes of DNA methylation and histone H3 methylation were determined. ATR upregulated the expression of ZC317.7, hsp-6, and hsp-60. Mitochondrial stress in parental generation dependent transcription 6mA modifiers may establish these epigenetic marks in progeny.

Keywords: ATR, Caenorhabditis elegans, multi-/trans-generation, reproductive toxicity

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391 Semi-Automatic Segmentation of Mitochondria on Transmission Electron Microscopy Images Using Live-Wire and Surface Dragging Methods

Authors: Mahdieh Farzin Asanjan, Erkan Unal Mumcuoglu

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Mitochondria are cytoplasmic organelles of the cell, which have a significant role in the variety of cellular metabolic functions. Mitochondria act as the power plants of the cell and are surrounded by two membranes. Significant morphological alterations are often due to changes in mitochondrial functions. A powerful technique in order to study the three-dimensional (3D) structure of mitochondria and its alterations in disease states is Electron microscope tomography. Detection of mitochondria in electron microscopy images due to the presence of various subcellular structures and imaging artifacts is a challenging problem. Another challenge is that each image typically contains more than one mitochondrion. Hand segmentation of mitochondria is tedious and time-consuming and also special knowledge about the mitochondria is needed. Fully automatic segmentation methods lead to over-segmentation and mitochondria are not segmented properly. Therefore, semi-automatic segmentation methods with minimum manual effort are required to edit the results of fully automatic segmentation methods. Here two editing tools were implemented by applying spline surface dragging and interactive live-wire segmentation tools. These editing tools were applied separately to the results of fully automatic segmentation. 3D extension of these tools was also studied and tested. Dice coefficients of 2D and 3D for surface dragging using splines were 0.93 and 0.92. This metric for 2D and 3D for live-wire method were 0.94 and 0.91 respectively. The root mean square symmetric surface distance values of 2D and 3D for surface dragging was measured as 0.69, 0.93. The same metrics for live-wire tool were 0.60 and 2.11. Comparing the results of these editing tools with the results of automatic segmentation method, it shows that these editing tools, led to better results and these results were more similar to ground truth image but the required time was higher than hand-segmentation time

Keywords: medical image segmentation, semi-automatic methods, transmission electron microscopy, surface dragging using splines, live-wire

Procedia PDF Downloads 142
390 Kauffman Model on a Network of Containers

Authors: Johannes J. Schneider, Mathias S. Weyland, Peter Eggenberger Hotz, William D. Jamieson, Oliver Castell, Alessia Faggian, Rudolf M. Füchslin

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In the description of the origin of life, there are still some open gaps, e.g., the formation of macromolecules cannot be fully explained so far. The Kauffman model proposes the existence of autocatalytic sets of macromolecules which mutually catalyze reactions leading to each other’s formation. Usually, this model is simulated in one well-stirred pot only, with a continuous inflow of small building blocks, from which larger molecules are created by a set of catalyzed ligation and cleavage reactions. This approach represents the picture of the primordial soup. However, the conditions on the early Earth must have differed geographically, leading to spatially different outcomes whether a specific reaction could be performed or not. Guided by this picture, the Kauffman model is simulated in a large number of containers in parallel, with neighboring containers being connected by diffusion. In each container, only a subset of the overall reaction set can be performed. Under specific conditions, this approach leads to a larger probability for the existence of an autocatalytic metabolism than in the original Kauffman model.

Keywords: agglomeration, autocatalytic set, differential equation, Kauffman model

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389 Metabolic Regulation of Rhizobacteria for Cool-Season Grass Tolerance to Heat Stress

Authors: Kashif Jaeel, Bingru Huang

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Stress-induced accumulation of ethylene exacerbates drought damages in plants, and suppressing stress induction of ethylene may promote plant tolerance to heat stress. The objective of this study was to investigate the effects of endophytic bacteria (Paraburkholderia aspalathi) with 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzymes in suppressing ethylene production on plant tolerance to heat stress and underlying physiological mechanisms of P. aspalathi-regulation in creeping bentgrass (Agrostis stolonifera). A novel strain of P. aspalathi, ‘WSF23’, with ACC deaminase activity was used to inoculate the roots of plants (cv. ‘Penncross’) subjected to heat stress in controlled-environment chambers. Inoculation with WSF23 bacteria resulted in improved shoot and root growth during heat stress. The differential changes in metabolite regulation due to the bacterial inoculation could contribute to ACC deamination bacteria-improved heat tolerance in cool-season grass species.

Keywords: rhizobacteria, grass, heat, plant metabolism, soil bacteria

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388 Phylogeographic Reconstruction of the Tiger Shrimp (Penaeus monodon) Invasion in the Atlantic Ocean: The Role of the Farming Systems in the Marine Biological Invasions

Authors: Juan Carlos Aguirre Pabon, Stephen Sabatino, James Morris, Khor Waiho, Antonio Murias

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The tiger shrimp Penaeus monodon is one of the most important species in aquaculture and is native to the Indo-Pacific Ocean. During its greatest success in world production (70s and 80s) was introduced in many Atlantic Ocean countries for cultivation purposes and is currently reported as established in several countries of this area. Because there are no studies to understand the magnitude of the invasion process, this is an exciting opportunity to test evolutionary hypotheses in the context of marine invasions mediated by culture systems; therefore, the purpose of this study was to reconstruct the scenario of invasion of P. monodon in the Atlantic Ocean, by using mitochondrial DNA and eight loci microsatellites. In addition, samples of the invasion area in the Atlantic Ocean (US, Colombia, Venezuela, Brazil, Guienne Bissau, Senegal), the Indo-Pacific Ocean (Indonesia, India, Mozambique), and some cultivation systems (India, Bangladesh, Madagascar) were collected; and analysis of phylogenetic relationships (using some species of the family), genetic diversity, structure population, and demographic changes were performed. High intraspecific divergence in P. semisulcatus and P. monodon were found, high genetic variability in all sites (especially with microsatellites) and the presence of three clusters or populations. In addition, signs of demographic expansion in the culture population and bottlenecks in the invasive and native populations were found, as well as evidence of gene mixtures from all of the populations studied, implying that cropping systems play an essential role in mitigating the negative effects of the founder effect and providing a source of genetic variability that can ensure the success of the invasion.

Keywords: species introduction, increased variability, demographic changes, promoting invasion.

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387 Synthesis and Anti-Inflammatory Activity of Pyrazol-3-yl Thiazole 4-Carboxylic Acid Derivatives Targeting Enzyme in the Leukotriene Pathway

Authors: Shweta Sinha, Mukesh Doble, Manju S. L.

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Pyrazole scaffold is an important group of compound in heterocyclic chemistry and is found to possess numerous uses in chemistry. Pyrazole derivatives are also known to possess important biological activities including antitumor, antimicrobial, antiviral, antifungal, anticancer and anti-inflammatory. Inflammation is associated with pain, allergy and asthma. Leukotrienes are mediators of various inflammatory and allergic disorders. 5-Lipoxygenase (5-LOX) is an important enzyme involved in the biosynthesis of leukotrienes and metabolism of arachidonic acid (AA) and thus targeted for anti-inflammation. In vitro inhibitory activity of pyrazol-3-yl thiazole 4-carboxylic acid derivatives is tested against enzyme 5-LOX. Most of these compounds exhibit good inhibitory activity against this enzyme. Binding mode study of these compounds is determined by computational tool. Further experiments are being done to understand the mechanism of action of these compounds in inhibiting this enzyme. To conclude, these compounds appear to be a promising target in drug design against 5-LOX.

Keywords: inflammation, inhibition, 5-lipoxygenase, pyrazole

Procedia PDF Downloads 218
386 Disruption of MoNUC1 Gene Mediates Conidiation in Magnaporthe oryzae

Authors: Irshad Ali Khan, Jian-Ping Lu, Xiao-Hong Liu, Fu-Cheng Lin

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This study reports the functional analysis of a gene MoNUC1 in M. oryzae, which is homologous to the Saccharomyces cerevisiae NUC1 encoding a mitochondrial nuclease protein. The MoNUC1 having a gene locus MGG_05324 is 1002-bp in length and encodes an identical protein of 333 amino acids. We disrupted the gene through gene disruption strategy and isolated two mutants confirmed by southern blotting. The deleted mutants were then used for phenotypic studies and their phenotypes were compared to those of the Guy-11 strain. The mutants were first grown on CM medium to find the effect of MoNUC1 gene disruption on colony growth and the mutants were found to show normal culture colony growth similar to that of the Guy-11 strain. Conidial germination and appressorial formation were also similar in both the mutants and Guy-11 strains showing that this gene plays no significant role in these phenotypes. For pathogenicity, the mutants and Guy-11 mycelium blocks were inoculated on blast susceptible barley seedlings and it was found that both the strains exhibited full pathogenicity showing coalesced and necrotic blast lesions suggesting that this gene is not involved in pathogenicity. Mating of the mutants with 2539 strain formed numerous perithecia showing that MoNUC1 is not essential for sexual reproduction in M. oryzae. However, the mutants were found to form reduced conidia (1.06±8.03B and 1.08±9.80B) than those of the Guy-11 strain (1.46±10.61A) and we conclude that this protein is not required for the blast fungus to cause pathogenicity but plays significant role in conidiation. Proteins of signal transduction pathways that could be disrupted/ intervened genetically or chemically could lead to antifungal products of important fungal cereal diseases and reduce rice yield losses. Tipping the balance toward understanding the whole of pathogenesis, rather than simply conidiation will take some time, but clearly presents the most exciting challenge of all.

Keywords: appressorium formation, conidiation, NUC1, Magnaporthe oryzae, pathogenicity

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385 A Review of Pharmacological Prevention of Peri-and Post-Procedural Myocardial Injury After Percutaneous Coronary Intervention

Authors: Syed Dawood Md. Taimur, Md. Hasanur Rahman, Syeda Fahmida Afrin, Farzana Islam

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The concept of myocardial injury, although first recognized from animal studies, is now recognized as a clinical phenomenon that may result in microvascular damage, no-reflow phenomenon, myocardial stunning, myocardial hibernation and ischemic preconditioning. The final consequence of this event is left ventricular (LV) systolic dysfunction leading to increased morbidity and mortality. The typical clinical case of reperfusion injury occurs in acute myocardial infarction (MI) with ST segment elevation in which an occlusion of a major epicardial coronary artery is followed by recanalization of the artery. This may occur either spontaneously or by means of thrombolysis and/or by primary percutaneous coronary intervention (PCI) with efficient platelet inhibition by aspirin (acetylsalicylic acid), clopidogrel and glycoprotein IIb/IIIa inhibitors. In recent years, percutaneous coronary intervention (PCI) has become a well-established technique for the treatment of coronary artery disease. PCI improves symptoms in patients with coronary artery disease and it has been increasing the safety of procedures. However, peri- and post-procedural myocardial injury, including angiographical slow coronary flow, microvascular embolization, and elevated levels of cardiac enzyme, such as creatine kinase and troponin-T and -I, has also been reported even in elective cases. Furthermore, myocardial reperfusion injury at the beginning of myocardial reperfusion, which causes tissue damage and cardiac dysfunction, may occur in cases of the acute coronary syndrome. Because patients with myocardial injury is related to larger myocardial infarction and have a worse long-term prognosis than those without myocardial injury, it is important to prevent myocardial injury during and/or after PCI in patients with coronary artery disease. To date, many studies have demonstrated that adjunctive pharmacological treatment suppresses myocardial injury and increases coronary blood flow during PCI procedures. In this review, we highlight the usefulness of pharmacological treatment in combination with PCI in attenuating myocardial injury in patients with coronary artery disease.

Keywords: coronary artery disease, percutaneous coronary intervention, myocardial injury, pharmacology

Procedia PDF Downloads 426
384 Genetic Diversity and Discovery of Unique SNPs in Five Country Cultivars of Sesamum indicum by Next-Generation Sequencing

Authors: Nam-Kuk Kim, Jin Kim, Soomin Park, Changhee Lee, Mijin Chu, Seong-Hun Lee

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In this study, we conducted whole genome re-sequencing of 10 cultivars originated from five countries including Korea, China, India, Pakistan and Ethiopia with Sesamum indicum (Zhongzho No. 13) genome as a reference. Almost 80% of the whole genome sequences of the reference genome could be covered by sequenced reads. Numerous SNP and InDel were detected by bioinformatic analysis. Among these variants, 266,051 SNPs were identified as unique to countries. Pakistan and Ethiopia had high densities of SNPs compared to other countries. Three main clusters (cluster 1: Korea, cluster 2: Pakistan and India, cluster 3: Ethiopia and China) were recovered by neighbor-joining analysis using all variants. Interestingly, some variants were detected in DGAT1 (diacylglycerol O-acyltransferase 1) and FADS (fatty acid desaturase) genes, which are known to be related with fatty acid synthesis and metabolism. These results can provide useful information to understand the regional characteristics and develop DNA markers for origin discrimination of sesame.

Keywords: Sesamum indicum, NGS, SNP, DNA marker

Procedia PDF Downloads 304
383 Cratoxy Formosum (Jack) Dyer Leaf Extract-Induced Human Breast and Liver Cancer Cells Death

Authors: Benjaporn Buranrat, Nootchanat Mairuae

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Cratoxylum formosum (Jack) Dyer (CF) has been used for the traditional medicines in South East Asian and Thailand. Normally, northeast Thai vegetables have proven cytotoxic to many cancer cells. Therefore, the present study aims to explore the molecular mechanisms underlying CF-induced cancer cell death and apoptosis on breast and liver cancer cells. The cytotoxicity and antiproliferative effects of CF on the human breast MCF-7 and liver HepG2 cancer cell lines were evaluated using sulforhodamine B assay and colony formation assay. Cell migration assay was measured using wound healing assay. The apoptosis induction mechanisms were investigated through reactive oxygen species formation, caspase 3 activity, and JC-1 activity. Gene expression by real-time PCR and apoptosis related protein levels by Western blot analysis. CF induced MCF-7 and HepG2 cell death by time- and dose-dependent manner. Furthermore, CF had the greater cytotoxic potency on MCF-7 more than HepG2 cells with IC50 values of 85.70+4.52 μM and 219.03±9.96 μM respectively, at 24 h. Treatment with CF also caused a dose-dependent decrease in colony forming ability and cell migration, especially on MCF-7 cells. CF induced ROS formation, increased caspase 3 activities, and decreased the mitochondrial membrane potential, and causing apoptotic body production and DNA fragmentation. CF significantly decreased expression of the cell cycle regulatory protein RAC1 and downstream proteins, cdk6. Additionally, CF enhanced p21 and reduced cyclin D1 protein levels. CF leaf extract induced cell death, apoptosis, antimigration in both of MCF-7 and HepG2 cells. CF could be useful for developing to anticancer drug candidate for breast and liver cancer therapy.

Keywords: cratoxylum formosum (jack) dyer, breast cancer, liver cancer, cell death

Procedia PDF Downloads 185