Search results for: milk proteins

Authors: Tatiana R. Simonyan, Elena A. Protasova, Anastasia V. Mamontova, Eugene G. Maksimov, Konstantin A. Lukyanov, Alexey M. Bogdanov

Abstract:

Here, we describe two variants of the calcium indicators based on the GCaMP sensitive core and BrUSLEE fluorescent protein (GCaMP-BrUSLEE and GCaMP-BrUSLEE-145). In contrast to the conventional GCaMP6-family indicators, these fluorophores are characterized by the well-marked responsiveness of their fluorescence decay kinetics to external calcium concentration both in vitro and in cellulo. Specifically, we show that the purified GCaMP-BrUSLEE and GCaMP-BrUSLEE-145 exhibit three-component fluorescence decay kinetics, with the amplitude-normalized lifetime component (t3*A3) of GCaMP-BrUSLEE-145 changing four-fold (500-2000 a.u.) in response to a Ca²⁺ concentration shift in the range of 0—350 nM. Time-resolved fluorescence microscopy of live cells displays the two-fold change of the GCaMP-BrUSLEE-145 mean lifetime upon histamine-stimulated calcium release. The aforementioned Ca²⁺-dependence calls considering the GCaMP-BrUSLEE-145 as a prospective Ca²⁺-indicator with the signal read-out in the time domain.

Keywords: calcium imaging, fluorescence lifetime imaging microscopy, fluorescent proteins, genetically encoded indicators

Procedia PDF Downloads 148

Authors: Mona Hajirasouli

Abstract:

Abstract: introduction: Antimicrobial compounds are important in the first line of the host defense system of many animal species. Material and methods: In the present study antibacterial activity of crude and proteins precipitate of hemolymph and crude hepatopancreas extracts from Portunus segnis and Grapsus albolineatus against a range of 6 different bacterial strains evaluated. Amoxicillin as a positive control were also used. Results: Maximum activity (15.9 mm) was recorded in male haemolymph of p.segnis against Entrobacter and minimum activity (7 mm) was recorded against Serratia marcescens, Enterobacter sp. and Proteus mirabilis from different extracts of Grapsus albolineatus. Data were analyzed using independent-t in SPSS version 16, and results indicate that there were not any significant differences between hemolymph and hepatopancreas extracts of 2 species. Discussion: Antimicrobial activity has been reported earlier in the hemolymph of some brachyuran crabs such as: blue crab Callinectes sapidus, mud crab Scylla serrata, Ocypode macrocera and Carcinus maenas. This study shows that hemolymph and hepatopancreas of Portunus segnis and Grapsus albolineatus may potential antibiotics.

Keywords: brachyuran, Portunus segnis, Grapsus albolineatus, hemolymph, hepatopancreas, antibacterial

Procedia PDF Downloads 162

Authors: Violina R. Angelova, Sava G. Tabakov, Aleksander B. Peltekov, Krasimir I. Ivanov

Abstract:

A field study was conducted to evaluate the chemical composition and quality of the Aronia fruits, as well as the possibilities of Aronia cultivation on soils contaminated with heavy metals. The experiment was performed on an agricultural field contaminated by the Non-Ferrous-Metal Works (NFMW) near Plovdiv, Bulgaria. The study included four varieties of Aronia; Aron variety, Hugin variety, Viking variety and Nero variety. The Aronia was cultivated according to the conventional technology on areas at a different distance from the source of pollution NFMW- Plovdiv (1 km, 3.5 km, and 15 km). The concentrations of macroelements, microelements, and heavy metals in Aronia fruits were determined. The dry matter content, ash, sugars, proteins, and fats were also determined. Aronia is a crop that is tolerant to heavy metals and can successfully be grown on soils contaminated with heavy metals. The increased content of heavy metals in the soil leads to less absorption of the nutrients (Ca, Mg and P) in the fruit of the Aronia. Soil pollution with heavy metals does not affect the quality of the Aronia fruit varieties.

Keywords: aronia, chemical composition, fruits, quality

Procedia PDF Downloads 192

Authors: Akshita Bhardwaj

Abstract:

Keratinase is an enzyme obtained from extracellular sources that is involved in biodegradation of keratin. It is a member of a group of proteases that can break down keratin into amino acids. Keratinases are produced only in the presence of substrate that contain keratin. It attacked the disulfide bond of substrate and involve in keratin degradation. Human hair, feathers, animal hard tissues, horns, claws, and hooves all contain keratin.. It exists in two form alpha keratin (found in soft tissues) and beta keratin (found in hard tissue). By taking part in the degradation of keratin, keratinases derived from microbial sources, often referred to as microbial keratinases, are important in the process of turning wastes containing keratin into products with added value. Chicken feathers contain high level of keratin protein content than other sources and became a suitable protein source. Keratinase production occurs at near alkaline pH and thermophilic temperatures. The bioprocessing of keratinous waste benefits greatly from the use of keratinases. Additionally, it lessens the issue caused by poultry excrement. The use of feather meal, along with keratinase, improves the digestion of proteins and amino acids.

Keywords: mili litre (ml), micro litre (Ul), TCA - trichloroacetic acid, OD - optical density

Procedia PDF Downloads 72

Authors: Ruolan Zhou

Abstract:

Trisomy of human chromosome 21 is the primary cause of Down Syndrome (DS), and this genetic disease has significantly burdened families and countries, causing great controversy. To address this problem, the research takes an approach in exploring the relationship between genetic abnormality and this disease's symptoms, adopting several techniques, including data analysis and enrichment analysis. It also explores open-source websites, such as NCBI, DAVID, SOURCE, STRING, as well as UCSC, to complement its result. This research has analyzed the variety of genes on human chromosome 21 with simple coding, and by using analysis, it has specified the protein-coding genes, their function, and their location. By using enrichment analysis, this paper has found the abundance of keratin production-related coding-proteins on human chromosome 21. By adopting past researches, this research has attempted to disclose the relationship between trisomy of human chromosome 21 and keratin production abnormality, which might be the reason for common diseases in patients with Down Syndrome. At last, by addressing the advantage and insufficiency of this research, the discussion has provided specific directions for future research.

Keywords: Down Syndrome, protein production, genome, enrichment analysis

Procedia PDF Downloads 115

Authors: Gianella Franco, Karen Suarez, María Quijano, Patricia Manzano

Abstract:

Agro-industries generate large amounts of waste, which are mostly untapped. This research was carried out to use cocoa, coffee and orange industrial by-products to develop a dairy drink. The product was prepared by making a 10% aqueous extract of the mixture of cocoa and coffee beans shells and orange peel. Extreme Vertices Mixture Design was applied to vary the proportions of the ingredients of the aqueous extract, getting 13 formulations. Each formulation was mixed with skim milk and pasteurized. The attributes of taste, smell, color and appearance were evaluated by a semi-trained panel by multiple comparisons test, comparing the formulations against a standard marked as "R", which consisted of a coffee commercial drink. The formulations with the highest scores were selected to maximize the Total Polyphenol Content (TPC) through a process of linear optimization resulting in the formulation 80.5%: 18.37%: 1.13% of cocoa bean shell, coffee bean shell and orange peel, respectively. The Total Polyphenol Content was 4.99 ± 0.34 mg GAE/g of drink, DPPH radical scavenging activity (%) was 80.14 ± 0.05 and caffeine concentration of 114.78 mg / L, while the coffee commercial drink presented 3.93 ± 0.84 mg GAE / g drink, 55.54 ± 0.03 % and 47.44 mg / L of TPC, DPPH radical scavenging activity and caffeine content, respectively. The results show that it is possible to prepare an antioxidant - rich drink with good sensorial attributes made of industrial by-products.

Keywords: DPPH, polyphenols, waste, food science

Procedia PDF Downloads 461

Authors: Mohana Babu Amberkar, Meena Kumari K, Ravi, Arjun, Christopher Rockson

Abstract:

The aim of this research is to evaluate the hepatoprotective activity of Terminalia paniculata (Tp) against ATT induced hepatic damage in rats.Three hepatotoxic ATT drugs Isoniazid + Rifampicin + Pyrazinamide, silymarin as standard hepatoprotective drug and 0.5% carboxymethylcellulose (CMC) as a control were used. Tp extract and silymarin were administered orally with ATT drugs for 90 days. Two doses 250 and 500 mg/kg of Tp extract, ATT drugs and silymarin were administered as suspensions with 0.5% CMC. ATT treated rats showed a significant increase in aspartate transaminase, alanine transaminase, alkaline phosphatase, lactate dehydrogenase, and lipid peroxides in the serum vs. control. Treatment of silymarin and Tp (250mg/kg) extract showed hepatoprotective activity against the hepatic damage by ATT. This was evident from significant reduction in serum liver enzymes levels, and also there was a significant increase in serum proteins, albumin and total liver tissue thiols as compared to the ATT treated groups. Tp was found to possess hepatoprotective property.

Keywords: antitubercular drugs, hepatoprotective, liver enzymes, Terminalia paniculata

Procedia PDF Downloads 429

Authors: Dubravka Štajner, Boris M. Popović, Saša Orlović, Ružica Ždero, Milan Popović, Aleksandra Popović

Abstract:

Many plants possess antioxidant ingredients that provides efficacy by additive or synergistic activities. Present article highlights an antioxidant capacity of Serbian conifer plants. Antioxidant activities of the crude extracts were assessed using different assays. In this study, quantities of phenolic compounds (total phenols, flavonoids, tannins and proanthocyanidins), contents of photosynthetic pigments (chlorophyll a and b and carotenoids), soluble proteins and proline were examined. MDA quantities and ability of extracts to remove reactive nitrogen and oxygen species (RNOS) were also investigated. Furthermore, antioxidant activities of extracts against DPPH∙, ferric reducing antioxidant power, permanganate reducing antioxidant capacity were also determined. According to almost all used assays, antioxidant and scavenging capacities of silver fir (Abies alba Mill.), and Douglas fir (Pseudotsuga menziesii) were superior compared to spruce. Presented results implicated that leaves of Douglas fir and silver fir possessed outstanding antioxidant characteristics that could diminish damage caused by oxygen radicals which are responsible for many of the bodily changes and susceptibility to different diseases.

Keywords: conifers, antioxidant activity, reducing power, lipid peroxidation

Procedia PDF Downloads 341

Authors: Jwan J. Abdullah, Greetham Darren, Gregory A, Tucker, Chenyu Du

Abstract:

Solid-state fermentation (SSF) is an alternative to liquid fermentations for the production of commercially important products such as antibiotics, single cell proteins, enzymes, organic acids, or biofuels from lignocellulosic material. This paper describes the optimisation of SSF on municipal solid waste (MSW) for the production of cellulase enzyme. Production of cellulase enzymes was optimised by Trichoderma reesei or Aspergillus niger for temperature, moisture content, inoculation, and period of incubation. Also, presence of minerals, and alternative carbon and nitrogen sources. Optimisation revealed that production of cellulolytic enzymes was optimal when using Trichoderma spp at 30°C with an incubation period of 168 hours with a 60% moisture content. Crude enzymes produced from MSW, by Trichoderma were evaluated for the saccharification of MSW and compared with activity of a commercially available enzyme, results demonstrated that MSW can be used as inexpensive lignocellulosic material for the production of cellulase enzymes using Trichoderma reesei.

Keywords: SSF, enzyme hydrolysis, municipal solid waste (MSW), optimizing conditions, enzyme hydrolysis

Procedia PDF Downloads 547

Authors: S. Ghalem, M. Mesmoudi, I. Daoudand, H. Allali

Abstract:

The nitrogen-containing bisphosphonates (N-BPs) are well established as the treatments of choice for disorders of excessive bone resorption, myeloma and bone metastases, and osteoporosis. They inhibit farnesyl pyrophosphate synthase (FFPS), a key enzyme in the mevalonate pathway, resulting in inhibition of the prenylation of small GTP-binding proteins in osteoclasts and disruption of their cytoskeleton, adhesion/spreading, and invasion of cancer cells. A very few examples for synthesis of α-amino bisphosphonates based on several amino acids are known from the literature. In the present work, esters of aminoacid react with ketophsophonate (or their analog acid or acyl) to afford the desired products, α-iminophosphonates. The reaction of imine with dimethyl phosphate in the presence of catalytic amount of I2 give ester of α-aminobisphosphonate as sole product in good yield. Finally, we used computational docking methods to predict how several α-aminobisphosphonates bind to FPPS and how R and X influence. Pamidronate, β-aminobisphosphonate already marketed, was used as reference. These results are of interest since they represent a new and simple way to sythesize α-aminobisphosphonates with a free COOH group increased by R2 functionalisable and opening up the possibility of using the molecular docking to facilitate the design of other, novel FFPS inhibitors.

Keywords: drug research, cancer, α-amino bisphosphonates, molecular docking

Procedia PDF Downloads 265

Authors: Abhilasha Dadhich, Manisha Mathur, Sanjay Kumar, Hemant Dadhich

Abstract:

The camel (Camelus dromedarius) is an important animal component of the fragile desert eco-system of India. Apart from others, impaired milk and meat production decrease in performance and even death are some of the major consequences of parasitic disease like coccidiosis in camel. Coccidiosis which is an acute invasion and destruction of intestinal mucosa by protozoa of the genera Eimeria or isospora spp. Post-Mortem examinations of 5 carcasses of dromedary of different age groups aged from 2 to 5 years were conducted. The history indicated that the camels were suffering from diarrhoea, dysentery, pyrexia, inappetence, weight loss, and emaciation. Post mortem examinations showed macroscopic and microscopic alterations in the small intestine, particularly in jejunum and ileum regions. The mucosae were congested, and haemorrhagic on which there were numerous whitish-grey nodular foci were observed. The affected intestinal tissue specimens were preserved in 10% formal saline and processed mechanically for paraffin embedding by acetone and benzene technique. The sections were stained with haematoxylin and eosin method of staining for histopathological examinations. Histologically, typical lesions such as congestion and haemorrhages were present. The intestinal villi were oedematous; mucosa degenerated and desquamated, along with infiltration of eosinophils and macrophages. Crypts of lieberkuhn were obliterated due to presence of schizonts in lamina propria. Older camels served as the source of spread of coccidial infection and were also predisposed to secondary infections.

Keywords: camel, coccidiosis, Eimeria, histopathology

Procedia PDF Downloads 182

Authors: Hamed A. El-Serehy, Khaled A. Al-Rasheid, Fahad A Al-Misned

Abstract:

The meiofaunal population fluctuation, microbial dynamic and the composition of the sedimentary organic matter were investigated seasonally in the Egyptian shores along the northern part of Red Sea. Total meiofaunal population densities were extremely low with an annual average of 109 ±26 ind./10 cm2 and largely dominated by nematodes (on annual average from 52% to 94% of total meiofaunal density). The benthic microbial population densities ranged from 0.26±0.02 x 108 to 102.67±18.62 x 108/g dry sediment. Total sedimentary organic matter concentrations varied between 5.8 and 11.6 mg/g and the organic carbon, which was measured as summation of the carbohydrates, proteins and lipids, accounted for only a small fraction of being 32 % of the total organic matter. Chlorophyll a attained very low values and fluctuated between 2 and 11 µg/g. The very low chlorophyll a concentration in the Egyptian coasts along the Red Sea can suggest that the sedimentary organic matter along the Egyptian coasts is dominated by organic detrital and heterotrophic bacteria on one hand, and do not promote carbon transfer towards the higher trophic level on the other hand. However, the present study indicates that the existing of well diversified meiofaunal group, with a total of ten meiofaunal taxa, can serve as food for higher trophic levels in the Red Sea marine ecosystem.

Keywords: bacteria, meiofauna, intertidal sediments, Red Sea

Procedia PDF Downloads 418

Authors: Khiem M. Nguyen, Ming C. Yang

Abstract:

Chloroplast pigments are extremely important during photosynthesis since they play essential roles in light absorption and energy transfer. Therefore, understanding the efficiency of chlorophyll (Chl) biosynthesis could facilitate enhancement in photo-assimilates accumulation, and ultimately, in crop yield. The Chl-deficient mutants have been used extensively to study the Chl biosynthetic pathways and the biogenesis of the photosynthetic apparatus. Rice (Oryza sativa L.) is one of the most leading food crops, serving as staple food for many parts of the world. To author’s best knowledge, Chl b–lacking rice has been found; however the molecular mechanism of Chl biosynthesis still remains unclear compared to wild-type rice. In this study, the ultrastructure analysis, photosynthetic properties, and transcriptome profile of wild-type rice (Norin No.8, N8) and its Chl b-lacking mutant (Chlorina 1, C1) were examined. The finding concluded that total Chl content and Chl b content in the C1 leaves were strongly reduced compared to N8 leaves, suggesting that reduction in the total Chl content contributes to leaf color variation at the physiological level. Plastid ultrastructure of C1 possessed abnormal thylakoid membranes with loss of starch granule, large number of vesicles, and numerous plastoglobuli. The C1 rice also exhibited thinner stacked grana, which was caused by a reduction in the number of thylakoid membranes per granum. Thus, the different Chl a/b ratio of C1 may reflect the abnormal plastid development and function. Transcriptional analysis identified 23 differentially expressed genes (DEGs) and 671 transcription factors (TFs) that were involved in Chl metabolism, chloroplast development, cell division, and photosynthesis. The transcriptome profile and DEGs revealed that the gene encoding PsbR (PSII core protein) was down-regulated, therefore suggesting that the lower in light-harvesting complex proteins are responsible for the lower photosynthetic capacity in C1. In addition, expression level of cell division protein (FtsZ) genes were significantly reduced in C1, causing chloroplast division defect. A total of 19 DEGs were identified based on KEGG pathway assignment involving Chl biosynthesis pathway. Among these DEGs, the GluTR gene was down-regulated, whereas the UROD, CPOX, and MgCH genes were up-regulated. Observation through qPCR suggested that later stages of Chl biosynthesis were enhanced in C1, whereas the early stages were inhibited. Plastid structure analysis together with transcriptomic analysis suggested that the Chl a/b ratio was amplified both by the reduction in Chl contents accumulation, owning to abnormal chloroplast development, and by the enhanced conversion of Chl b to Chl a. Moreover, the results indicated the same Chl-cycle pattern in the wild-type and C1 rice, indicating another Chl b degradation pathway. Furthermore, the results demonstrated that normal grana stacking, along with the absence of Chl b and greatly reduced levels of Chl a in C1, provide evidence to support the conclusion that other factors along with LHCII proteins are involved in grana stacking. The findings of this study provide insight into the molecular mechanisms that underlie different Chl a/b ratios in rice.

Keywords: Chl-deficient mutant, grana stacked, photosynthesis, RNA-Seq, transcriptomic analysis

Procedia PDF Downloads 111

Authors: Ragy Raafat Gaber Attaalla

Abstract:

PIP: This review, based on 2 large-scale studies, discusses the pharmacology and physiology of oral steroid contraceptives (OCs). The pharmacological distinction between synthetic and naturally occurring steroids centers on changes in biological activity dependent on compound formulation and an individual's metabolism. OC mechanism of action is explained as the main prevention of ovulation by interference with gonadotropin-releasing hormone. Since some 52 metabolic alterations have been reported in OC users, these phenomena are dealt with in 3 categories: 1) effects on the primary target organs of the female reproductive tract (ovary, myometrium, endometrium, cervix, vagina, breasts, and hypothalamus), 2) general metabolic effects (serum proteins, carbohydrate metabolism, lipid metabolism, water and electrolyte metabolism, body weight, tryptophan metabolism, and vitamins and minerals), and 3) effects on other organ systems (liver, central nervous system, skin, genitourinary, gastrointestinal tract, eye, immune phenomena, and effect on subsequent fertility). The choice of the proper OC formulation and use of OCs by adolescents are discussed. Assessment of OC safety, contraindications, and patient monitoring are provided.

Keywords: steroid oral contraceptives, ovulation, female reproductive tract, metabolic effects

Procedia PDF Downloads 91

Authors: Hassan Ali Abood Nassrullah, Jabbar Fadeel Mahdi, Mohammed Salih Mahdi Alkurdi, Ali Al Mousawi, Saad Ibrahim Al-Ghabban, Abdul Amir H. Kadhum, Ahmed Al-Amiery

Abstract:

Background: Q fever is a zoonotic disease characterized by its clinical polymorphism and can present acutely as fever, pneumonia, hepatitis, and chronically as infective endocarditis, arthritis, osteomyelitis, or hepatitis. Objective: The aim of this study is To estimate the prevalence of cases of Q fever in hospitalized febrile patients in Imam Al Hussain Teaching Medical City in Karbala. Methods: One hundred patients with pyrexia were admitted to the medical ward from 1st August to 31st December 2019. Serological procedures fortified by Enzyme-linked Immunosorbent Assay test. Patients were considered to have acute Q fever when the specific antibodies (IgM and IgG) of phase II of Coxiella burnetii were positive. Results: The mean age of the patients was 35.05±12.93 years; females constituted 60% of them. Eighteen patients (18%) showed positive results for IgM, a lower proportion (13% n=13) had positive IgG levels, and 9% showed equivocal results. Statistical analysis revealed a significant association between positive IgM levels of the female gender and in patients consuming unpasteurized milk. One patient (female aged 60 years) died in the hospital, while all other patients were discharged well. Two female patients were pregnant, and one of them had an abortion. Conclusions: Q fever is more common in febrile patients. The study indicates that this disease should not be overlooked in the differential diagnosis of acute fever. Serological testing should be performed in all patients with acute febrile illness with an unsettling diagnosis.

Keywords: antibodies, frequency, immunoglobulin IgM, Q fever

Procedia PDF Downloads 114

Authors: Phuriwat Laomethakorn, Siritron Samosorn, Ramida Watanapokasin

Abstract:

Cancer metastasis is the major cause of cancer-related death. Therefore searching for a compound that could inhibit cancer metastasis is necessary. 13-Butoxyberberine bromide is a berberine derivative that has not been reported an anti-metastatic effect on skin cancer cells. This study aimed to investigate the anti-metastatic effect of 13-butoxyberberine bromide on skin cancer A431 cells. The effect of 13-butoxyberberine bromide on A431 cell viability was examined by MTT assay. Suppression of cell migration and invasion in A431 cells were determined by wound healing assay, transwell migration assay, and transwell invasion assay. Metastasis proteins were determined by western blotting. The results demonstrated that 13-butoxyberberine bromide decreased A431 cell viability in a dose-dependent manner. In addition, sub-toxic concentrations of 13-butoxyberberine bromide suppressed cell migration and invasion in A431 cells. In addition, 13-butoxyberberine bromide showed anti-metastatic effects by down-regulated MMP-2 and MMP-9 expression. These findings may be useful in the development of 13-butoxyberberine bromide as an anti-metastatic drug in the future.

Keywords: 13-butoxyberberine bromide, metastasis, skin cancer, MMP

Procedia PDF Downloads 98

Authors: Victor Arokia Doss, Kuberapandian Dharaniyambigai, K. Julia Rose Mary

Abstract:

Advanced Glycation End (AGE) products are the end products due to the reaction between excess reducing sugar present in diabetes and free amino group in protein lipids and nucleic acids. Thus, non-enzymic glycation of molecules such as hemoglobin, collagen, and other structurally and functionally important proteins add to the pathogenic complications such as diabetic retinopathy, neuropathy, nephropathy, vascular changes, atherosclerosis, Alzheimer's disease, rheumatoid arthritis, and chronic heart failure. The most common non-cross linking AGE, carboxymethyl lysine (CML) is formed by the oxidative breakdown of fructosyllysine, which is a product of glucose and lysine. CML is formed in a wide variety of tissues and is an index to assess the extent of glycoxidative damage. Thus we have constructed a mathematical and computational model that predicts the effect of temperature differences in vivo, on the formation of CML, which is now being considered as an important intracellular milieu. This hybrid model that had been tested for its parameter fitting and its sensitivity with available experimental data paves the way for designing novel laboratory experiments that would throw more light on the pathological formation of AGE adducts and in the pathophysiology of diabetic complications.

Keywords: advanced glycation end-products, CML, mathematical model, computational model

Procedia PDF Downloads 127

Authors: Mohd Sidek Ahmad, Zainon Mohd Noor, Zaidah Zainal Ariffin

Abstract:

Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Keywords: isolation, identification, fibrinolytic protease, endophytic fungi, Hibiscus leaves

Procedia PDF Downloads 425

Authors: K. Żak, S. Przetocka, R. Kitel, K. Guzik, B. Musielak, S. Malicki, G. Dubin, T. A. Holak

Abstract:

Studies on tumor genesis revealed a number of factors that may potentially serve as molecular targets for immunotherapies. One of such promising targets are PD1 and PDL1 proteins. PD1 (Programmed cell death protein 1) is expressed by activated T cells and plays a critical role in modulation of the host's immune response. One of the PD1 ligands -PDL1- is expressed by macrophages, monocytes and cancer cells which exploit it to avoid immune attack. The notion of the mechanisms used by cancer cells to block the immune system response was utilized in the development of therapies blocking PD1-PDL1 interaction. Up to date, human PD1-PDL1 complex has not been crystallized and structure of the mouse-human complex does not provide a complete view of the molecular basis of PD1-PDL1 interactions. The purpose of this study is to obtain crystal structure of the human PD1-PDL1 complex which shall allow rational design of small molecule inhibitors of the interaction. In addition, the study presents results of binding small-molecules to PD1 and fragment docking towards PD1 protein which will facilitate the design and development of small–molecule inhibitors of PD1-PDL1 interaction.

Keywords: PD1, PDL1, cancer, small molecule, drug discovery

Procedia PDF Downloads 390

Authors: Marina Passero, Tianhua Zhai, Zuyi (Jacky) Huang

Abstract:

Alzheimer’s disease has become a major public health issue, as indicated by the increasing populations of Americans living with Alzheimer’s disease. After decades of extensive research in Alzheimer’s disease, only seven drugs have been approved by Food and Drug Administration (FDA) to treat Alzheimer’s disease. Five of these drugs were designed to treat the dementia symptoms, and only two drugs (i.e., Aducanumab and Lecanemab) target the progression of Alzheimer’s disease, especially the accumulation of amyloid-b plaques. However, controversial comments were raised for the accelerated approvals of either Aducanumab or Lecanemab, especially with concerns on safety and side effects of these two drugs. There is still an urgent need for further drug discovery to target the biological processes involved in the progression of Alzheimer’s disease. Excessive cholesterol has been found to accumulate in the brain of those with Alzheimer’s disease. Cholesterol can be synthesized in both the blood and the brain, but the majority of biosynthesis in the adult brain takes place in astrocytes and is then transported to the neurons via ApoE. The blood brain barrier separates cholesterol metabolism in the brain from the rest of the body. Various proteins contribute to the metabolism of cholesterol in the brain, which offer potential targets for Alzheimer’s treatment. In the astrocytes, SREBP cleavage-activating protein (SCAP) binds to Sterol Regulatory Element-binding Protein 2 (SREBP2) in order to transport the complex from the endoplasmic reticulum to the Golgi apparatus. Cholesterol is secreted out of the astrocytes by ATP-Binding Cassette A1 (ABCA1) transporter. Lipoprotein receptors such as triggering receptor expressed on myeloid cells 2 (TREM2) internalize cholesterol into the microglia, while lipoprotein receptors such as Low-density lipoprotein receptor-related protein 1 (LRP1) internalize cholesterol into the neuron. Cytochrome P450 Family 46 Subfamily A Member 1 (CYP46A1) converts excess cholesterol to 24S-hydroxycholesterol (24S-OHC). Cholesterol has been approved for its direct effect on the production of amyloid-beta and tau proteins. The addition of cholesterol to the brain promotes the activity of beta-site amyloid precursor protein cleaving enzyme 1 (BACE1), secretase, and amyloid precursor protein (APP), which all aid in amyloid-beta production. The reduction of cholesterol esters in the brain have been found to reduce phosphorylated tau levels in mice. In this work, a computational pipeline was developed to identify the protein targets involved in cholesterol regulation in brain and further to identify chemical compounds as the inhibitors of a selected protein target. Since extensive evidence shows the strong correlation between brain cholesterol regulation and Alzheimer’s disease, a detailed literature review on genes or pathways related to the brain cholesterol synthesis and regulation was first conducted in this work. An interaction network was then built for those genes so that the top gene targets were identified. The involvement of these genes in Alzheimer’s disease progression was discussed, which was followed by the investigation of existing clinical trials for those targets. A ligand-protein docking program was finally developed to screen 1.5 million chemical compounds for the selected protein target. A machine learning program was developed to evaluate and predict the binding interaction between chemical compounds and the protein target. The results from this work pave the way for further drug discovery to regulate brain cholesterol to combat Alzheimer’s disease.

Keywords: Alzheimer’s disease, drug discovery, ligand-protein docking, gene-network analysis, cholesterol regulation

Procedia PDF Downloads 65

Authors: Saleh Gorji, Sahil Gulati, Ana Pakzad

Abstract:

Continuous electron diffraction tomography, also known as microcrystal electron diffraction (MicroED) or three-dimensional electron diffraction (3DED), is a powerful technique, which in combination with cryo-electron microscopy (cryo-ED), can provide atomic-scale 3D information about the crystal structure and composition of different classes of crystalline materials such as proteins, peptides, and small molecules. Unlike the well-established X-ray crystallography method, 3DED does not require large single crystals and can collect accurate electron diffraction data from crystals as small as 50 – 100 nm. This is a critical advantage as growing larger crystals, as required by X-ray crystallography methods, is often very difficult, time-consuming, and expensive. In most cases, specimens studied via 3DED method are electron beam sensitive, which means there is a limitation on the maximum amount of electron dose one can use to collect the required data for a high-resolution structure determination. Therefore, collecting data using a conventional scintillator-based fiber coupled camera brings additional challenges. This is because of the inherent noise introduced during the electron-to-photon conversion in the scintillator and transfer of light via the fibers to the sensor, which results in a poor signal-to-noise ratio and requires a relatively higher and commonly specimen-damaging electron dose rates, especially for protein crystals. As in other cryo-EM techniques, damage to the specimen can be mitigated if a direct detection camera is used which provides a high signal-to-noise ratio at low electron doses. In this work, we have used two classes of such detectors from Gatan, namely the K3® camera (a monolithic active pixel sensor) and Stela™ (that utilizes DECTRIS hybrid-pixel technology), to address this problem. The K3 is an electron counting detector optimized for low-dose applications (like structural biology cryo-EM), and Stela is also a counting electron detector but optimized for diffraction applications with high speed and high dynamic range. Lastly, data collection workflows, including crystal screening, microscope optics setup (for imaging and diffraction), stage height adjustment at each crystal position, and tomogram acquisition, can be one of the other challenges of the 3DED technique. Traditionally this has been all done manually or in a partly automated fashion using open-source software and scripting, requiring long hours on the microscope (extra cost) and extensive user interaction with the system. We have recently introduced Latitude® D in DigitalMicrograph® software, which is compatible with all pre- and post-energy-filter Gatan cameras and enables 3DED data acquisition in an automated and optimized fashion. Higher quality 3DED data enables structure determination with higher confidence, while automated workflows allow these to be completed considerably faster than before. Using multiple examples, this work will demonstrate how to direct detection electron counting cameras enhance 3DED results (3 to better than 1 Angstrom) for protein and small molecule structure determination. We will also show how Latitude D software facilitates collecting such data in an integrated and fully automated user interface.

Keywords: continuous electron diffraction tomography, direct detection, diffraction, Latitude D, Digitalmicrograph, proteins, small molecules

Procedia PDF Downloads 99

Authors: Marcela Menah de Sousa Lima, Victor Ushijima Leone, Natasha Aparecida Grande de Franca, Barbara Santarosa Emo Peters, Ligia Araujo Martini

Abstract:

Conjugated linoleic acid (CLA) intake has been constantly related to benefits to human health since having a positive effect on reducing body fat. The aim of the present study was to investigate the association between CLA intake and biochemical measurements and body composition of adults and the elderly. Subjects/Methods: 287 adults and elderly participants in an epidemiological study in Sao Paulo Brazil, were included in the present study. Participants had their dietary data obtained by two non-consecutive 24HR, a body composition assessed by dual-energy absorptiometry exam (DXA), and a blood collection. Mean differences and a correlation test was performed. For all statistical tests, a significance of 5% was considered. Results: CLA intake showed a positive correlation with HDL-c levels (r = 0.149; p = 0.011) and negative with VLDL-c levels (r = -0.134; p = 0.023), triglycerides (r = -0.135; p = 0.023) and glycemia (r = -0.171; p = 0.004), as well as negative correlation with visceral adipose tissue (VAT) (r = -0.124, p = 0.036). Evaluating individuals in two groups according to VAT values, a significant difference in CLA intake was observed (p = 0.041), being the group with the highest VAT values, the one with the lowest fatty acid intake. Conclusions: This study suggests that CLA intake is associated with a better lipid profile and lower visceral adipose tissue volume, which contributes to the investigation of the effects of CLA on obesity parameters. However, it is necessary to investigate the effects of CLA from milk and dairy products in the control adiposity.

Keywords: adiposity, dairy products, diet, fatty acids

Procedia PDF Downloads 133

Authors: Saimir Heta, Ilma Robo, Dhimiter Papakozma, Eduart Kapaj, Vera Ostreni

Abstract:

Introduction: The biocompatible materials applied to the implant surfaces are the target of recent literature studies. Methodologies: Modification of implant surfaces in different ways such as application of additional ions, surface microstructure change, surface or laser ultrasound alteration, or application of various substances such as recombinant proteins are among the most affected by articles published in the literature. The study is of review type with the main aim of finding the different ways that the mesenchymal cell reaction to these materials is, according to the literature, in the same percentage positive to the osteointegration process. Results: It is emphasized in the literature that implant success as a key evaluation key has more to implement implant treatment protocol ranging from dental health amenity and subsequent of the choice of implant type depending on the alveolar shape of the ridge level. Conclusions: Osteointegration is a procedure that should initially be physiologically independent of the type of implant pile material. With this physiological process, it can not "boast" for implant success or implantation depending on the brand of the selected implant, as the breadth of synthetic or natural materials that promote osteointegration is relatively large.

Keywords: mesenchymal cells, implants, review, biocompatible materials

Procedia PDF Downloads 83

Authors: Eman Alzahrani

Abstract:

Efficient extraction of proteins by removing interfering materials is necessary in proteomics, since most instruments cannot handle such contaminated sample matrices directly. In this study, chitosan-coated magnetic nanoparticles (CS-MNPs) for purification of myoglobin were successfully fabricated. First, chitosan (CS) was prepared by a deacetylation reaction during its extraction from shrimp-shell waste. Second, magnetic nanoparticles (MNPs) were synthesised, using the coprecipitation method, from aqueous Fe2+ and Fe3+ salt solutions by the addition of a base under an inert atmosphere, followed by modification of the surface of MNPs with chitosan. The morphology of the formed nanoparticles, which were about 23 nm in average diameter, was observed by transmission electron microscopy (TEM). In addition, nanoparticles were characterised using X-ray diffraction patterns (XRD), which showed the naked magnetic nanoparticles have a spinel structure and the surface modification did not result in phase change of the Fe3O4. The coating of MNPs was also demonstrated by scanning electron microscopy (SEM) analysis, energy dispersive analysis of X-ray spectroscopy (EDAX), and Fourier transform infrared (FT-IR) spectroscopy. The adsorption behaviour of MNPs and CS-MNPs towards myoglobin was investigated. It was found that the difference in adsorption capacity between MNPs and CS-MNPs was larger for CS-MNPs. This result makes CS-MNPs good adsorbents and attractive for using in protein extraction from biological samples.

Keywords: chitosan, magnetic nanoparticles, coprecipitation, adsorption

Procedia PDF Downloads 412

Authors: H. Chassaigne, S. Gioria, J. Lobo Vicente, D. Carpi, P. Barboro, G. Tomasi, A. Kinsner-Ovaskainen, F. Rossi

Abstract:

Emerging approaches in the area of exposure to nanomaterials and assessment of human health effects combine the use of in vitro systems and analytical techniques to study the perturbation of the proteome and/or the metabolome. We investigated the modification in the cytoplasmic compartment of the Balb/3T3 cell line exposed to gold nanoparticles. On one hand, the proteomic approach is quite standardized even if it requires precautions when dealing with in vitro systems. On the other hand, metabolomic analysis is challenging due to the chemical diversity of cellular metabolites that complicate data elaboration and interpretation. Differentially expressed proteins were found to cover a range of functions including stress response, cell metabolism, cell growth and cytoskeleton organization. In addition, de-regulated metabolites were annotated using the HMDB database. The "omics" fields hold huge promises in the interaction of nanoparticles with biological systems. The combination of proteomics and metabolomics data is possible however challenging.

Keywords: data processing, gold nanoparticles, in vitro systems, metabolomics, proteomics

Procedia PDF Downloads 495

Authors: F. R. Chen, C. Kisielowski, D. Van Dyck

Abstract:

In principle, the latest generation aberration-corrected transmission electron microscopes (TEMs) could achieve sub-Å resolution, but there is bottleneck that hinders the final step towards revealing 3D structure. Firstly, in order to achieve a resolution around 1 Å with single atom sensitivity, the electron dose rate needs to be sufficiently large (10⁴-10⁵eÅ⁻² s⁻¹). With such large dose rate, the electron beam can induce surfaces alterations or even bulk modifications, in particular, for electron beam sensitive (soft) materials such as nm size particles, organic materials, proteins or macro-molecules. We will demonstrate methodology of low dose electron holography for observing 3D structure for soft materials such as single Oleic acid molecules at atomic resolution. The main improvement of this new type of electron holography is based on two concepts. Firstly, the total electron dose is distributed over many images obtained at different defocus values from which the electron hologram is then reconstructed. Secondly, in contrast to the current tomographic methods that require projections from several directions, the 3D structural information of the nano-object is then extracted from this one hologram obtained from only one viewing direction.

Keywords: low dose electron microscopy, in-line electron holography, atomic resolution tomography, soft materials

Procedia PDF Downloads 183

Authors: V. Viswanathan, Md. Irshad Ahmad, Prashant K. Singh, Nayeem Ahmad, Pradeep Sharma, Sujata Sharma, Tej P Singh

Abstract:

Lactoperoxidase (LPO) is a heme containing mammalian enzyme which uses hydrogen peroxide (H2O2) to catalyze the conversion of substrates into oxidized products. LPO is found in body fluids and tissues such as milk, saliva, tears, mucosa and other body secretions. The previous structural studies have shown that LPO converts substrates, thiocyanate (SCN‾) and iodide (I‾) ions into oxidized products, hypothiocyanite (OSCN‾) and hypoiodite (IO‾) ions, respectively. We report here a new structure of the complex of LPO with an oxidized product, nitrite (NO2‾). This product was generated from NO using the two step reaction of LPO by adding hydrogen peroxide (H2O2) in the solution of LPO in 0.1M phosphate buffer at pH 6.8 as the first step. In the second step, NO gas was added to the above mixture. This was crystallized using 20% (w/v) PEG-3350 and 0.2M ammonium iodide at pH 6.8. The structure determination showed the presence of NO2‾ ion in the distal heme cavity of the substrate binding site of LPO. The structure also showed that the propionate group, which is linked to pyrrole ring D of the heme moiety, was disordered. Similarly, the side chain of Asp108, which is covalently linked to heme moiety, was also split into two components. As a result of these changes, the conformation of the side chain of Arg255 was altered, allowing it to form new interactions with the disordered carboxylic group of propionate moiety. These structural changes are indicative of an intermediate state in the catalytic reaction pathway of LPO.

Keywords: lactoperoxidase, structure, nitric oxide, nitrite ion, intermediate, complex

Procedia PDF Downloads 96

Authors: Yu-Ching Huang

Abstract:

Background: Delivery bridges the antepartum and postpartum period. Subsequent fatigue can affect indices, including postpartum recovery and life quality. Milk secretion, breastfeeding quality, and newborn participation may be compromised. Correspondingly, using proper pushing skills during the second stage of labor has the potential to effectively reduce postpartum fatigue and enhance birth satisfaction in new mothers. Purpose: To compare the effects of using different pushing skills on maternal pushing perception, postpartum fatigue, and birth satisfaction. Methodology: The present study used a descriptive research approach and recruited 382 participants from a medical center in northern Taiwan. Data were collected using a structured questionnaire, which included a demographic and obstetrics information datasheet, the Labor Pushing Experience Scale, a fatigue scale, and a birth satisfaction scale. Research Results: Using pushing skills (including upright position [t= 2.28, p < .05] and delayed pushing [t= -1.98, p < .05] during the second stage of labor was shown to enhance birth satisfaction in participants. Additionally, open glottis pushing ( t = 5.46, p < .001) resulted in a mean duration of second-stage labor that was 17.67 minutes less than that achieved using Valsalva pushing. Moreover, a better perceived pushing experience was associated with lower perceived postpartum fatigue (r = .46, p < .05) and higher birth satisfaction (r = -.16, p < .05). Finally, postpartum fatigue perception was negatively associated with birth satisfaction (r = -.16, p < .05). Conclusion and Clinical Application: The findings suggest that midwives should advocate that women adopt upright positions, delayed pushing, and open glottis pushing during the second stage of labor in order to enhance their birth satisfaction.

Keywords: second stage labor duration of pushing skill, pushing experience perception, postpartum fatigue, birth satisfaction

Procedia PDF Downloads 263

Authors: Muhammad Ali Syed, Arshad Saleem Bhatti, Chen-zhong Li, Habib Ali Bokhari

Abstract:

Surface plasmon resonance (SPR) biosensors have emerged as a promising technique for bioanalysis as well as microbial detection and identification. Real time, sensitive, cost effective, and label free detection of biomolecules from complex samples is required for early and accurate diagnosis of infectious diseases. Like many other types of optical techniques, SPR biosensors may also be successfully utilized for microbial detection for accurate, point of care, and rapid results. In the present study, we have utilized a commercially available automated SPR biosensor of BI company to study the microbial detection form water samples spiked with different concentration of Staphylococcus aureus bacterial cells. The gold thin film sensor surface was functionalized to react with proteins such as protein G, which was used for directed immobilization of monoclonal antibodies against Staphylococcus aureus. The results of our work reveal that this immunosensor can be used to detect very small number of bacterial cells with higher sensitivity and specificity. In our case 10^3 cells/ml of water have been successfully detected. Therefore, it may be concluded that this technique has a strong potential to be used in microbial detection and identification.

Keywords: surface plasmon resonance (SPR), Staphylococcus aureus, biosensors, microbial detection

Procedia PDF Downloads 469

Authors: Hilal Ahmad Punoo

Abstract:

Conjugated linoleic acid (CLA) is a mixture of positional and geometric isomers of octadecadienoic acid with two conjugated double bonds. Of more than a dozen isomers of CLA found naturally in dairy and meat products from ruminants, c-9, t-11 and t-10, c-12 are the two isomers with known physiological importance, including anticarcinogenic, antidiabetic, antilipogenic, and antiatherosclerotic effects. Conjugated linoleic acids (CLA) may influence the onset and severity of several chronic diseases, including various cancers, atherosclerosis, obesity, bone density loss, and diabetes. These findings are of special interest to the agriculture community, because dietary sources of CLA are almost exclusively beef and dairy products. Thus, a better understanding of the specific isomers and mechanisms responsible for these positive effects of CLA on human health would be both prudent and economically beneficial. To date, research related to the advantages of CLA consumption on human health has been conducted using experimental laboratory animals and cell culture systems. These data consistently show that relatively low levels of CLA can influence risk of cancer. Further, very recent investigations suggest that the predominate CLA isoform (cis-9, trans-11 CLA or rumenic acid) found in beef and milk fat possesses anticarcinogenic effects but does not alter body composition; the trans-10, cis-12 CLA has been shown to inhibit lipogenesis. Clearly, further work, especially using human subjects, will be required to characterize the potential benefits of CLA consumption on human health. Moreover, we suggest that foods naturally containing high amounts of CLA (e.g., beef and dairy products) be considered as meeting the definition of functional foods.

Keywords: conjugated linoleic acid, potential health benefits, fats, animals, humans

Procedia PDF Downloads 302