Search results for: pollen proteome
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 90

Search results for: pollen proteome

90 Common Ragweed (Ambrosia artemisiifolia): Changing Proteomic Patterns of Pollen under Elevated NO₂ Concentration and/or Future Rising Temperature Scenario

Authors: Xiaojie Cheng, Ulrike Frank, Feng Zhao, Karin Pritsch

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Ragweed (Ambrosia artemisiifolia) is an invasive weed that has become an increasing global problem. In addition to affecting land use and crop yields, ragweed has a strong impact on human health as it produces highly allergenic pollen. Global warming will result in an earlier and longer pollen season enhanced pollen production and an increase in pollen allergenicity with a negative effect on atopic patients. The aims of this study were to investigate the effects of increasing temperature, the future climate scenario in the Munich area, southern Germany, predicted on the basis of RCP8.5 until the end of 2050s, or/and NO₂, a major air pollutant, 1) on the vegetative and reproductive characteristics of ragweed plants, 2) on the total allergenicity of ragweed pollen, 3) on the total pollen proteomic patterns. Ragweed plants were cultivated for the whole plant vegetation period under controlled conditions either under ambient climate conditions or 4°C higher temperatures with or without additional NO₂. Higher temperature resulted in bigger plant sizes, longer male inflorescences, and longer pollen seasons. The total allergenic potential of the pollen was accessed by dot blot using serum from ragweed pollen sensitized patients. The comparative immunoblot analysis revealed that the in vivo fumigation of ragweed plants with elevated NO₂-concentrations significantly increased the allergenic potential of the pollen, and in combination with increased temperature, the allergenic potential was even higher. On the other hand, label-free protein quantification by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed. The results showed that more proteins were significantly up- and down-regulated under higher temperatures with/without elevated NO₂ conditions. Most of the highly expressed proteins were participating intensively in the metabolic process, the cellular process, and the stress defense process. These findings suggest that rising temperature and elevated NO₂ are important environmental factors for higher abiotic stress activities, catalytic activities, and thus higher allergenic potential observed in pollen proteins.

Keywords: climate change, NO₂, pollen proteome, ragweed, temperature

Procedia PDF Downloads 147
89 Logistic Model Tree and Expectation-Maximization for Pollen Recognition and Grouping

Authors: Endrick Barnacin, Jean-Luc Henry, Jack Molinié, Jimmy Nagau, Hélène Delatte, Gérard Lebreton

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Palynology is a field of interest for many disciplines. It has multiple applications such as chronological dating, climatology, allergy treatment, and even honey characterization. Unfortunately, the analysis of a pollen slide is a complicated and time-consuming task that requires the intervention of experts in the field, which is becoming increasingly rare due to economic and social conditions. So, the automation of this task is a necessity. Pollen slides analysis is mainly a visual process as it is carried out with the naked eye. That is the reason why a primary method to automate palynology is the use of digital image processing. This method presents the lowest cost and has relatively good accuracy in pollen retrieval. In this work, we propose a system combining recognition and grouping of pollen. It consists of using a Logistic Model Tree to classify pollen already known by the proposed system while detecting any unknown species. Then, the unknown pollen species are divided using a cluster-based approach. Success rates for the recognition of known species have been achieved, and automated clustering seems to be a promising approach.

Keywords: pollen recognition, logistic model tree, expectation-maximization, local binary pattern

Procedia PDF Downloads 150
88 Development of Allergenic and Melliferous Floral Pollen Spectrum Using Scanning Electron Microscopy

Authors: Mehwish Jamil Noor

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Morphological features of pollen (sculpturing) were useful for identification of different floral taxa. In this study 49 pollen grains, types belonging to 25 families were studied using Scanning Electron Microscope. Shape and sculpturing of pollen ranging from Psilate, scabrate to reticulate, bireticulate and echinolophate. Honey pollen was identified using morphological features, number and arrangement of pore and colpi, size and shape. It presents the first attempt from Pakistan involving extraction of pollen from honey, its identification and taxonomic analysis. Among pollen studied diversity in shape and sculpturing has been observed ranging from Psilate, scabrate to reticulate to bireticulate and echinolophate condition. Pollen has been identified with the help of morphological feature, number and arrangement of pore and colpi, size and shape, reference slides, light microscopic data and previous literature have been consulted for pollen identification. Pollen of closely related species resemble each other therefore pollen identification of airborne and honey pollen is not possible till species level. Survey of flora was carried in parallel to keep the record about the allergenic and melliferous preference of specific sites through surveys and interviews. Their pollination season and geographical distribution were recorded. Two hundred and five including wild and cultivated taxa were identified belonging to sixty-seven families. Major bee attracting wild shrub and trees includes Justicia adhatoda, Acacia nilotica, Ziziphus jujuba, Taraxicum officinalis, Artemisia dubia, Casuarina sp., Ulmus sp., Broussonetia papyrifera, Cupressus sp. or Pinus roxburghii etc. Cultivated crops like Pennisetum typhoides, Nigella sativa, Triticum sativum along with fruit trees of Pyrus, Prunus, Eryobotria, Citrus etc. are popular melliferous floras. Exotic/ introduced species like Eucalyptus or Parthenium hysterophorus, are also frequently visited by bees indicating the significance of those plants in the honey industry. It is concluded that different microscopic analysis techniques give more clear and authentic pictures of and melliferous pollen identification which is well supported by the floral calendar. The diversity of pollen are observed in case of melliferous pollen, and most of the windborne pollen were found less sculptured or psilate expressing the adaptation to the specific mode of pollination. Pollen morphology and sculpturing would serve as a reference for future studies.

Keywords: pollen, allergenic flora, sem, pollen key, Scanning Electron Microscopy (SEM)

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87 Impact of Climate Variability on Dispersal and Distribution of Airborne Pollen and Fungal Spores in Nsukka, South-East Nigeria: Implication on Public Health

Authors: Dimphna Ezikanyi, Gloria Sakwari

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Airborne pollen and fungal spores are major triggers of allergies, and their abundance and seasonality depend on plant responses to climatic and meteorological variables. A survey of seasonal prevalence of airborne pollen and fungal spores in Nsukka, Enugu, South- East Nigeria and relationship to climatic variables were carried out from Jan-June, 2017. The aim of the study was to access climate change and variability over time in the area and their accrued influence on modern pollen and spores rain. Decadal change in climate was accessed from variables collected from meteorological centre in the study area. Airborne samples were collected monthly using a modified Tauber-like pollen samplers raised 5 ft above ground level. Aerosamples collected were subjected to acetolysis. Dominant pollen recorded were those of Poaceae, Elaeis guinensis Jacq. and Casuarina equisetifolia L. Change in weather brought by onset of rainfall evoked sporulation and dispersal of diverse spores into ambient air especially potent allergenic spores with the spores of Ovularia, Bispora, Curvularia, Nigrospora, Helminthosporium preponderant; these 'hydrophilic fungi' were abundant in the rainy season though in varying quantities. Total fungal spores correlated positively with monthly rainfall and humidity but negatively with temperature. There was a negative though not significant correlation between total pollen count and rainfall. The study revealed a strong influence of climatic variables on abundance and spatial distribution of pollen and fungal spores in the ambient atmosphere.

Keywords: allergy, fungal spores, pollen, weather parameters

Procedia PDF Downloads 142
86 Profile of Cross-Reactivity Allergens Highlighted by Multiplex Technology “Alex Microchip Technique” in the Diagnosis of Type I Hypersensitivity

Authors: Gadiri Sabiha

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Introduction: Current allergy diagnostic tools using Multiplex technology have made it possible to increase the efficiency of the search for specific IgE. This opportunity is provided by the newly developed “Alex Biochip”, consisting of a panel of 282 allergens in native and molecular form, a CCD inhibitor, and the potential for detecting cross-reactive allergens. We evaluated the performance of this technology in detecting cross-reactivity in previously explored patients. Material/Method: The sera of 39 patients presenting sensitization and polysensitization profiles were explored. The search for specific IgE is carried out by the Alex ® IgE Biochip, and the results are analyzed by nature and by molecular family of allergens using specific software. Results/Discussion: The analysis gave a particular profile of cross-reactivity allergens: 33% for the Ole e1 family, 31% for NPC2, 26% for storage proteins, 20% for Tropomyosin, 10% for LTPs, 10% for Arginine Kinase and 10% for Uteroglobin CCDs were absent in all patients. The “Ole e1” allergen is responsible for a pollen-pollen cross allergy. The storage proteins found and LTP are not species-specific, causing cross-pollen-food allergy. The nDer p2 of the NPC2 family is responsible for cross-reactivity between mite species. Conclusion: The cross-reactivities responsible for mixed syndromes at diagnosis in our patients were dominated by pollen-pollen and pollen-food syndromes. They allow the identification of severity factors linked to the prognosis and the best-adapted immunotherapy.

Keywords: specific IgE, allergy, cross reactivity, molecular allergens

Procedia PDF Downloads 34
85 Microorganisms in Fresh and Stored Bee Pollen Originated from Slovakia

Authors: Vladimíra Kňazovická, Mária Dovičičová, Miroslava Kačániová, Margita Čanigová

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The aim of the study was to test the storage of bee pollen at room temperature and in cold store, and to describe microorganisms originated from it. Fresh bee pollen originating in West Slovakia was collected in May 2010. It was tested for presence of particular microbial groups using dilution plating method, and divided into two parts with different storage (in cold store and at room temperature). Microbial analyses of pollen were repeated after one year of storage. Several bacterial strains were isolated and tested using Gram staining, for catalase and fructose-6-phosphate-phosphoketolase presence, and by rapid ID 32A (BioMérieux, France). Micromycetes were identified at genus level. Fresh pollen contained coliform bacteria, which were not detected after one year of storage in both ways. Total plate count (TPC) of aerobes and anaerobes and of yeasts in fresh bee pollen exceeded 5.00 log CFU/g. TPC of aerobes and anaerobes decreased below 2.00 log CFU/g after one year of storage in both ways. Count of yeasts decreased to 2.32 log CFU/g (at room temperature) and to 3.66 log CFU/g (in cold store). Microscopic filamentous fungi decreased from 3.41 log CFU/g (fresh bee pollen) to 1.13 log CFU/g (at room temperature) and to 1.89 log CFU/g (in cold store). In fresh bee pollen, 12 genera of micromycetes were identified in the following order according to their relative density: Penicillium > Mucor > Absidia > Cladosporium, Fusarium > Alternaria > Eurotium > Aspergillus, Rhizopus > Emericella > Arthrinium and Mycelium sterilium. After one year at room temperature, only three genera were detected in bee pollen (Penicillium > Aspergillus, Mucor) and after one year in cold store, seven genera were detected (Mucor > Penicillium, Emericella > Aspergillus, Absidia > Arthrinium, Eurotium). From the plates designated for anaerobes, eight colonies originating in fresh bee pollen were isolated. Among them, a single yeast isolate occurred. Other isolates were G+ bacteria, with a total of five rod shaped. In three out of these five, catalase was absent and fructose-6-phosphate-phosphoketolase was present. Bacterial isolates originating in fresh pollen belonged probably to genus Bifidobacterium or relative genera, but their identity was not confirmed unequivocally. In general, cold conditions are suitable for maintaining the natural properties of foodstuffs for a longer time. Slight decrease of microscopic fungal number and diversity was recorded in cold temperatures compared with storage at room temperature.

Keywords: bacteria, bee product, microscopic fungi, biosystems engineering

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84 Transcriptional Response of Honey Bee to Differential Nutritional Status and Nosema Infection

Authors: Farida Azzouz-Olden, Arthur G. Hunt, Gloria Degrandi-Hoffman

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Bees are confronting several environmental challenges, including the intermingled effects of malnutrition and disease. Intuitively, pollen is the healthiest nutritional choice; however, commercial substitutes, such as BeePro and MegaBee, are widely used. Herein we examined how feeding natural and artificial diets shapes transcription in the abdomen of the honey bee, and how transcription shifts in combination with Nosema parasitism. Gene ontology enrichment revealed that, compared with poor diet (carbohydrates (C)), bees fed pollen (P > C), BeePro (B > C), and MegaBee (M > C) showed a broad upregulation of metabolic processes, especially lipids; however, pollen feeding promoted more functions and superior proteolysis. The superiority of the pollen diet was also evident through the remarkable overexpression of vitellogenin in bees fed pollen instead of MegaBee or BeePro. Upregulation of bioprocesses under carbohydrates feeding compared to pollen (C > P) provided a clear poor nutritional status, uncovering stark expression changes that were slight or absent relatively to BeePro (C > B) or MegaBee (C > M). Poor diet feeding (C > P) induced starvation response genes and hippo signaling pathway, while it repressed growth through different mechanisms. Carbohydrate feeding (C > P) also elicited ‘adult behavior’, and developmental processes suggesting transition to foraging. Finally, it altered the ‘circadian rhythm’, reflecting the role of this mechanism in the adaptation to nutritional stress in mammals. Nosema-infected bees fed pollen compared to carbohydrates (PN > CN) upheld certain bioprocesses of uninfected bees (P > C). Poor nutritional status was more apparent against pollen (CN > PN) than BeePro (CN > BN) or MegaBee (CN > MN). Nosema accentuated the effects of malnutrition since more starvation-response genes and stress response mechanisms were upregulated in CN > PN compared to C > P. The bioprocess ‘Macromolecular complex assembly’ was also enriched in CN > PN, and involved genes associated with human HIV and/or influenza, thus providing potential candidates for bee-Nosema interactions. Finally, the enzyme Duox emerged as essential for guts defense in bees, similarly to Drosophila. These results provide evidence of the superior nutritional status of bees fed pollen instead of artificial substitutes in terms of overall health, even in the presence of a pathogen.

Keywords: honeybee, immunity, Nosema, nutrition, RNA-seq

Procedia PDF Downloads 125
83 Comparison of Machine Learning and Deep Learning Algorithms for Automatic Classification of 80 Different Pollen Species

Authors: Endrick Barnacin, Jean-Luc Henry, Jimmy Nagau, Jack Molinie

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Palynology is a field of interest in many disciplines due to its multiple applications: chronological dating, climatology, allergy treatment, and honey characterization. Unfortunately, the analysis of a pollen slide is a complicated and time consuming task that requires the intervention of experts in the field, which are becoming increasingly rare due to economic and social conditions. That is why the need for automation of this task is urgent. A lot of studies have investigated the subject using different standard image processing descriptors and sometimes hand-crafted ones.In this work, we make a comparative study between classical feature extraction methods (Shape, GLCM, LBP, and others) and Deep Learning (CNN, Autoencoders, Transfer Learning) to perform a recognition task over 80 regional pollen species. It has been found that the use of Transfer Learning seems to be more precise than the other approaches

Keywords: pollens identification, features extraction, pollens classification, automated palynology

Procedia PDF Downloads 95
82 Aerofloral Studies and Allergenicity Potentials of Dominant Atmospheric Pollen Types at Some Locations in Northwestern Nigeria

Authors: Olugbenga S. Alebiosu, Olusola H. Adekanmbi, Oluwatoyin T. Ogundipe

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Pollen and spores have been identified as major airborne bio-particles inducing respiratory disorders such as asthma, allergic rhinitis and atopic dermatitis among hypersensitive individuals. An aeropalynological study was conducted within a one year sampling period with a view to investigating the monthly depositional rate of atmospheric pollen and spores; influence of the immediate vegetation on airborne pollen distribution; allergenic potentials of dominant atmospheric pollen types at selected study locations in Bauchi and Taraba states, Northwestern Nigeria. A tauber-like pollen trap was employed in aerosampling with the sampler positioned at a height of 5 feet above the ground, followed by a monthly collection of the recipient solution for the sampling period. The collected samples were subjected to acetolysis treatment, examined microscopically with the identification of pollen grains and spores using reference materials and published photomicrographs. Plants within the surrounding vegetation were enumerated. Crude protein contents extracted from pollen types found to be commonly dominant at both study locations; Senna siamea, Terminalia cattapa, Panicum maximum and Zea mays were used to sensitize Musmusculus. Histopathological studies of bronchi and lung sections from certain dead M.musculus in the test groups was conducted. Blood samples were collected from the pre-orbital vein of M.musculus and processed for serological and haematological (differential and total white blood cell counts) studies. ELISA was used in determining the levels of serological parameters: IgE and cytokines (TNF-, IL-5, and IL-13). Statistical significance was observed in the correlation between the levels of serological and haematological parameters elicited by each test group, differences between the levels of serological and haematological parameters elicited by each test group and those of the control, as well as at varying sensitization periods. The results from this study revealed dominant airborne pollen types across the study locations; Syzygiumguineense, Tridaxprocumbens, Elaeisguineensis, Mimosa sp., Borreria sp., Terminalia sp., Senna sp. and Poaceae. Nephrolepis sp., Pteris sp. and a trilete fern also produced spores. This study also revealed that some of the airborne pollen types were produced by local plants at the study locations. Bronchi sections of M.musculus after first and second sensitizations, as well as lung section after first sensitization with Senna siamea, showed areas of necrosis. Statistical significance was recorded in the correlation between the levels of some serological and haematological parameters produced by each test group and those of the control, as well as at certain sensitization periods. The study revealed some candidate pollen allergens at the study locations allergy sufferers and also established a complexity of interaction between immune cells, IgE and cytokines at varied periods of mice sensitization and forming a paradigm of human immune response to different pollen allergens. However, it is expedient that further studies should be conducted on these candidate pollen allergens for their allergenicity potential in humans within their immediate environment.

Keywords: airborne, hypersensitive, mus musculus, pollen allergens, respiratory, tauber-like

Procedia PDF Downloads 105
81 Some Probiotic Traits of Lactobacillus Strains Isolated from Pollen

Authors: Hani Belhadj, Daoud Harzallah, Seddik Khennouf, Saliha Dahamna, Mouloud Ghadbane

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In this study, Lactobacillus strains isolated from pollen were identified by means of phenotypic and genotypic methods, At pH 2, most strains proved to be acid resistants, with losses in cell viability ranging from 0.77 to 4.04 Log orders. In addition, at pH 3 all strains could grew and resist the acidic conditions, with losses in cell viability ranging from 0.40 to 3.61 Log orders. It seems that, 0.3% and 0.5% of bile salts does not affect greatly the survival of most strains, excluding Lactobacillus sp. BH1398. Survival ranged from 81.0±3.5 to 93.5±3.9%. In contrast, in the presence of 1.0% bile salts, survival of five strains was decreased by more than 50%. Lactobacillus fermentum BH1509 was considered the most tolerant strain (77.5% for 1% bile) followed by Lactobacillus plantarum BH1541 (59.9% for 1% bile). Furthermore, all strains were resistant to colistine, clindamycine, chloramphenicol, and ciprofloxacine, but most of the strains were susceptible to Peniciline, Oxacillin, Oxytetracyclin, and Amoxicillin. Functionally interesting Lactobacillus isolates may be used in the future as probiotic cultures for manufacturing fermented foods and as bioactive delivery systems.

Keywords: probiotics, lactobacillus, pollen, bile, acid tolerance

Procedia PDF Downloads 389
80 Changing Colours and Odours: Exploring Cues Used by Insect Pollinators in Two Brassicaceous Plants

Authors: Katherine Y. Barragan-Fonseca, Joop J. A. Van Loon, Marcel Dicke, Dani Lucas-Barbosa

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Flowering plants use different traits to attract pollinators, which indicate flower location and reward quality. Visual and olfactory cues are among the most important floral traits exploited by pollinating insects. Pollination can alter physical and chemical cues of flowers, which can subsequently influence the behaviour of flower visitors. We investigated the main cues exploited by the syrphid fly Episyrphus balteatus and the butterfly Pieris brassicae when visiting flowers of Brassica nigra and Raphanus sativus plants. We studied post-pollination changes and their effects on the behaviour of flower visitors and flower volatile emission. Preference of pollinators was investigated by offering visual and olfactory cues simultaneously as well as separately in two-choice bioassays. We also assessed whether pollen is used as a cue by pollinating insects. In addition, we studied whether behavioural responses could be correlated with changes in plant volatile emission, by collecting volatiles from flower headspace. P. brassicae and E. balteatus did not use pollen as a cue in either of the two plant species studied. Interestingly, pollinators showed a strong bias for visual cues over olfactory cues when exposed to B. nigra plants. Flower visits by pollinators were influenced by post-pollination changes in B. nigra. In contrast, plant responses to pollination did not influence pollinator preference for R. sativus flowers. These results correlate well with floral volatile emission of B. nigra and R. sativus; pollination influenced the volatile profile of B. nigra flowers but not that of R. sativus. Collectively, our data show that different pollinators exploit different visual and olfactory traits when searching for nectar or pollen of flowers of two close related plant species. Although the syrphid fly consumes mostly pollen from brassicaceous flowers, it cannot detect pollen from a distance and likely associates other flower traits with quantity and quality of pollen.

Keywords: plant volatiles, pollinators, post-pollination changes, visual and odour cues

Procedia PDF Downloads 124
79 Dynamic Cardiac Mitochondrial Proteome Alterations after Ischemic Preconditioning

Authors: Abdelbary Prince, Said Moussa, Hyungkyu Kim, Eman Gouda, Jin Han

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We compared the dynamic alterations of mitochondrial proteome of control, ischemia-reperfusion (IR) and ischemic preconditioned (IPC) rabbit hearts. Using 2-DE, we identified 29 mitochondrial proteins that were differentially expressed in the IR heart compared with the control and IPC hearts. For two of the spots, the expression patterns were confirmed by Western blotting analysis. These proteins included succinate dehydrogenase complex, Acyl-CoA dehydrogenase, carnitine acetyltransferase, dihydrolipoamide dehydrogenase, Atpase, ATP synthase, dihydrolipoamide succinyltransferase, ubiquinol-cytochrome c reductase, translation elongation factor, acyl-CoA dehydrogenase, actin alpha, succinyl-CoA Ligase, dihydrolipoamide S-succinyltransferase, citrate synthase, acetyl-Coenzyme A dehydrogenase, creatine kinase, isocitrate dehydrogenase, pyruvate dehydrogenase, prohibitin, NADH dehydrogenase (ubiquinone) Fe-S protein, enoyl Coenzyme A hydratase, superoxide dismutase [Mn], and 24-kDa subunit of complex I. Interestingly, most of these proteins are associated with the mitochondrial respiratory chain, antioxidant enzyme system, and energy metabolism. The results provide clues as to the cardioprotective mechanism of ischemic preconditioning at the protein level and may serve as potential biomarkers for detection of ischemia-induced cardiac injury.

Keywords: ischemic preconditioning, mitochondria, proteome, cardioprotection

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78 Experimental and Modelling Performances of a Sustainable Integrated System of Conditioning for Bee-Pollen

Authors: Andrés Durán, Brian Castellanos, Marta Quicazán, Carlos Zuluaga-Domínguez

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Bee-pollen is an apicultural-derived food product, with a growing appreciation among consumers given the remarkable nutritional and functional composition, in particular, protein (24%), dietary fiber (15%), phenols (15 – 20 GAE/g) and carotenoids (600 – 900 µg/g). These properties are given by the geographical and climatic characteristics of the region where it is collected. There are several countries recognized by their pollen production, e.g. China, United States, Japan, Spain, among others. Beekeepers use traps in the entrance of the hive where bee-pollen is collected. After the removal of foreign particles and drying, this product is ready to be marketed. However, in countries located along the equator, the absence of seasons and a constant tropical climate throughout the year favors a more rapid spoilage condition for foods with elevated water activity. The climatic conditions also trigger the proliferation of microorganisms and insects. This, added to the factor that beekeepers usually do not have adequate processing systems for bee-pollen, leads to deficiencies in the quality and safety of the product. In contrast, the Andean region of South America, lying on equator, typically has a high production of bee-pollen of up to 36 kg/year/hive, being four times higher than in countries with marked seasons. This region is also located in altitudes superior to 2500 meters above sea level, having extremes sun ultraviolet radiation all year long. As a mechanism of defense of radiation, plants produce more secondary metabolites acting as antioxidant agents, hence, plant products such as bee-pollen contain remarkable more phenolics and carotenoids than collected in other places. Considering this, the improvement of bee-pollen processing facilities by technical modifications and the implementation of an integrated cleaning and drying system for the product in an apiary in the area was proposed. The beehives were modified through the installation of alternative bee-pollen traps to avoid sources of contamination. The processing facility was modified according to considerations of Good Manufacturing Practices, implementing the combined use of a cabin dryer with temperature control and forced airflow and a greenhouse-type solar drying system. Additionally, for the separation of impurities, a cyclone type system was implemented, complementary to a screening equipment. With these modifications, a decrease in the content of impurities and the microbiological load of bee-pollen was seen from the first stages, principally with a reduction of the presence of molds and yeasts and in the number of foreign animal origin impurities. The use of the greenhouse solar dryer integrated to the cabin dryer allowed the processing of larger quantities of product with shorter waiting times in storage, reaching a moisture content of about 6% and a water activity lower than 0.6, being appropriate for the conservation of bee-pollen. Additionally, the contents of functional or nutritional compounds were not affected, even observing an increase of up to 25% in phenols content and a non-significant decrease in carotenoids content and antioxidant activity.

Keywords: beekeeping, drying, food processing, food safety

Procedia PDF Downloads 79
77 Proteomics Application in Disease Diagnosis and Reproduction İmprovement in Cow

Authors: Abdollah Sobhani, Hossein Vaseghi-Dodaran

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Proteomics is defined as the study of the component of a cell, tissue and biological fluid. This technique has the potential to identify protein biomarkers of a disease states. In this study which was performed on bovine ovarian follicular cysts (BOFC), eight proteins are over expressed in BOFC that these proteins could be useful biomarkers for BOFC. The difference between serum proteome pattern cows affected by postpartum endometritis with healthy cows revealed that concentrations orosomucoid was decreased in endometritis. The comparison proteome of brucella abortus between laboratory adapted strains and clinical isolates could be useful to better understand this disease and vaccine development. Proteomics experiments identified new proteins and pathways that may be important in future hypothesis-driven studies of glucocorticoid-induced immunosuppression. Understanding the molecular mechanisms of effective parameters on male fertility is essential for obtaining high reproductive efficiency by decreasing cost and time. The investigations on proteome of high fertility spermatozoa indicated that expression of some proteins such as casein kinase 2 (CKII) prime poly peptide and tyrosine kinase in high fertility spermatozoa was higher compared to low fertility spermatozoa. Also, some evidence has indicated that variation in protein types and amounts in seminal fluid regulates fertility indexes in dairy bull. In conclusion, proteomics is a useful technique for discovering drugs, vaccine development, and diagnosis disease by biomarkers and improvement of reproduction efficiency.

Keywords: proteomics, reproduction, biomarker, immunity

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76 Palyno-Morphological Characteristics of Gymnosperm Flora of Pakistan and Its Taxonomic Implications with Light Microscope and Scanning Electron Microscopy Methods

Authors: Raees Khan, Sheikh Z. Ul Abidin, Abdul S. Mumtaz, Jie Liu

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The present study is intended to assess gymnosperms pollen flora of Pakistan using Light Microscope (LM) and Scanning Electron Microscopy (SEM) for its taxonomic significance in identification of gymnosperms. Pollens of 35 gymnosperm species (12 genera and five families) were collected from its various distributional sites of gymnosperms in Pakistan. LM and SEM were used to investigate different palyno-morphological characteristics. Five pollen types (i.e., Inaperturate, Monolete, Monoporate, Vesiculate-bisaccate, and Polyplicate) were observed. In equatorial view seven types of pollens were observed, in which ten species were sub-angular, nine species were triangular, six species were perprolate, three species were rhomboidal, three species were semi-angular, two species were rectangular and two species were prolate. While five types of pollen were observed in polar view, in which ten species were spheroidal, nine species were angular, eight were interlobate, six species were circular, and two species were elliptic. Eighteen species have rugulate and 17 species has faveolate ornamentation. Eighteen species have verrucate and 17 have gemmate type sculpturing. The data was analysed through cluster analysis. The study showed that these palyno-morphological features have significance value in classification and identification of gymnosperms. Based on these different palyno-morphological features, a taxonomic key was proposed for the accurate and fast identifications of gymnosperms from Pakistan.

Keywords: gymnosperms, palynology, Pakistan, taxonomy

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75 CMPD: Cancer Mutant Proteome Database

Authors: Po-Jung Huang, Chi-Ching Lee, Bertrand Chin-Ming Tan, Yuan-Ming Yeh, Julie Lichieh Chu, Tin-Wen Chen, Cheng-Yang Lee, Ruei-Chi Gan, Hsuan Liu, Petrus Tang

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Whole-exome sequencing focuses on the protein coding regions of disease/cancer associated genes based on a priori knowledge is the most cost-effective method to study the association between genetic alterations and disease. Recent advances in high throughput sequencing technologies and proteomic techniques has provided an opportunity to integrate genomics and proteomics, allowing readily detectable mutated peptides corresponding to mutated genes. Since sequence database search is the most widely used method for protein identification using Mass spectrometry (MS)-based proteomics technology, a mutant proteome database is required to better approximate the real protein pool to improve disease-associated mutated protein identification. Large-scale whole exome/genome sequencing studies were launched by National Cancer Institute (NCI), Broad Institute, and The Cancer Genome Atlas (TCGA), which provide not only a comprehensive report on the analysis of coding variants in diverse samples cell lines but a invaluable resource for extensive research community. No existing database is available for the collection of mutant protein sequences related to the identified variants in these studies. CMPD is designed to address this issue, serving as a bridge between genomic data and proteomic studies and focusing on protein sequence-altering variations originated from both germline and cancer-associated somatic variations.

Keywords: TCGA, cancer, mutant, proteome

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74 Fatty Acid Composition and Therapeutic Effects of Beebread

Authors: Sibel Silici

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Palynological spectrum, proximate and fatty acids composition of eight beebread samples obtained from different geographical origins were determined. Beebread moisture contents varied between 11.4-15.9 %, ash 1.9-2.54 %, fat 5.9-11.5 %, and protein between 14.8-24.3 %. To our knowledge, this is the first study investigating fatty acids (FAs) composition of the selected monofloral beebreads. A total of thirty-seven FAs were identified. Of these (9Z, 12Z, 15Z)-octadeca-9, 12, 15-trienoic acid, (9Z, 12Z)-octadeca-9, 12-dienoic acid, hexadecanoic acid, (Z)-octadec-9-enoic acid, (Z)-icos-11-enoic acid and octadecanoic acid were the most abundant in all the samples. Cotton beebread contained the highest level of ω-3 FAs, 41.3 %. Unsaturated/saturated FAs ratios ranged between 1.38 and 2.39 indicating that beebread is a good source of unsaturated FAs. The pollen, proximate and FAs composition of beebread samples of different botanical and geographical origins varied significantly.

Keywords: bee bread, fatty acid composition, proximate composition, pollen analysis

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73 Palynological Investigation and Quality Determination of Honeys from Some Apiaries in Northern Nigeria

Authors: Alebiosu Olugbenga Shadrak, Victor Victoria

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Honey bees exhibit preferences in their foraging behaviour on pollen and nectar for food and honey production, respectively. Melissopalynology is the study of pollen in honey and other honey products. Several work have been conducted on the palynological studies of honeys from the southern parts of Nigeria but with relatively scant records from the Northern region of the country. This present study aimed at revealing the favourably visited plants by honey bees, Apis melifera var. adansonii, at some apiaries in Northern Nigeria, as well as determining the quality of honeys produced. Honeys were harvested and collected from four apiaries of the region, namely: Sarkin Dawa missionary bee farm, Taraba State; Eleeshuwa Bee Farm, Keffi, Nassarawa State, Bulus Beekeeper Apiaries, Kagarko, Kaduna State and Mai Gwava Bee Farm, Kano State. These honeys were acetolysed for palynological microscopic analysis and subjected to standard treatment methods for the determination of their proximate composition and sugar profiling. Fresh anthers of two dominantly represented plants in the honeys were then collected for the quantification of their pollen protein contents, using the micro-kjeldhal procedure. A total of 30 pollen types were identified in the four honeys, and some of them were common to the honeys. A classification method for expressing pollen frequency class was employed: Senna cf. siamea, Terminalia cf. catappa, Mangifera indica, Parinari curatelifolia, Vitellaria paradoxa, Elaeis guineensis, Parkia biglobosa, Phyllantus muellerianus and Berlina Grandiflora, as “Frequent” (16-45%); while the others are either Rare (3-15%) or Sporadic (less than 3 %). Pollen protein levels of the two abundantly represented plants, Senna siamea (15.90mg/ml) and Terminalia catappa (17.33mg/ml) were found to be considerably lower. The biochemical analyses revealed varying amounts of proximate composition, non-reducing sugar and total sugar levels in the honeys. The results of this study indicate that pollen and nectar of the “Frequent” plants were preferentially foraged by honeybees in the apiaries. The estimated pollen protein contents of Senna same and Terminalia catappa were considerably lower and not likely to have influenced their favourable visitation by honeybees. However, a relatively higher representation of Senna cf. siamea in the pollen spectrum might have resulted from its characteristic brightly coloured and well scented flowers, aiding greater entomophily. Terminalia catappa, Mangifera indica, Elaeis guineensis, Vitellaria paradoxa, and Parkia biglobosa are typical food crops; hence they probably attracted the honeybees owing to the rich nutritional values of their fruits and seeds. Another possible reason for a greater entomophily of the favourably visited plants are certain nutritional constituents of their pollen and nectar, which were not investigated in this study. The nutritional composition of the honeys was observed to fall within the safe limits of international norms, as prescribed by Codex Alimentarius Commission, thus they are good honeys for human consumption. It is therefore imperative to adopt strategic conservation steps in ensuring that these favourably visited plants are protected from indiscriminate anthropogenic activities and also encourage apiarists in the country to establish their bee farms more proximally to the plants for optimal honey yield.

Keywords: honeybees, melissopalynology, preferentially foraged, nutritional, bee farms, proximally

Procedia PDF Downloads 241
72 Differential Response of Cellular Antioxidants and Proteome Expression to Salt, Cadmium and Their Combination in Spinach (Spinacia oleracea)

Authors: Rita Bagheri, Javed Ahmed, Humayra Bashir, M. Irfan Qureshi

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Agriculture lands suffer from a combination of stresses such as salinity and metal contamination including cadmium at the same time. Under such condition of multiple stresses, plant may exhibit unique responses different from the stress occurring individually. Thus, it would be interesting to investigate that how plant respond to combined stress at level of antioxidants and proteome expression, and identifying the proteins which are involved in imparting stress tolerance. With an approach of comparative proteomics and antioxidant analysis, present study investigates the response of Spinacia oleracea to salt (NaCl), cadmium (Cd), and their combination (NaCl+Cd) stress. Two-dimensional gel electrophoresis was used for resolving leaf proteome, and proteins of interest were identified using PDQuest software. A number of proteins expressed differentially, those indicated towards their roles in imparting stress tolerance, were digested by trypsin and analyzed on mass spectrometer for peptide mass fingerprinting (PMF). Data signals were then matched with protein databases using MASCOT. Results show that NaCl, Cd and both together (NaCl+Cd) induce oxidative stress which was highest in combined stress of Cd+NaCl. Correspondingly, the activities of enzymatic antioxidants viz., SOD, APX, GR and CAT, and non-enzymatic antioxidants had highest changes under combined stress compares to single stress over their respective controls. Among the identified proteins, several interesting proteins were identified that may be have role in Spinacia oleracia tolerance in individual and combinatorial stress of salt and cadmium. The functional classification of identified proteins indicates the importance and necessity of keeping higher ratio of defence and disease responsive proteins.

Keywords: Spinacia oleracea, Cd, salinity, proteomics, antioxidants, combinatorial stress

Procedia PDF Downloads 349
71 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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70 Functional Expression and Characterization of a Novel Indigenous Endo-Beta 1,4- Glucanase from Apis mellifera

Authors: Amtul Jamil Sami

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Apis mellifera is an insect of immense economic importance lives on rich carbohydrate diet including cellulose, nectar, honey and pollen. The carbohydrate metabolism in A mellifera has not been understood fully, as there are no data available, on the functional expression of cellulase gene. The cellulose hydrolyzing enzyme is required for the digestion of pollen cellulose wall, to release the important nutrients (amino acids, minerals, vitamins etc.) from the pollen. A dissection of Apis genome had revealed that there is one gene present for the expression of endo-beta-1,4-glucanase, for cellulose hydrolysis. In the presented work, functional expression of endo-beta-1,4 glucanase gene is reported. Total soluble proteins of the honey bee were isolated and were tested cellulose hydrolyzing enzyme activity, using carboxy-methyl cellulose, as a substrate. A mellifera proteins were able to hydrolyze carboxy-methyl cellulose, confirming its endo- type mode of action. Endo beta-1,4 glucanase enzyme was only present in the gut tissues, no activity was detected in the salivary glands. The pH optima of the enzyme were in the acidic pH range of 4-5-5-0, indicating its metabolic role in the acidic stomach of A mellifera. The reported enzyme is unique, as endo-beta- 1,4 glucanase was able to generate non reducing sugar, as an end product. The results presented, are supportive to the information that the honey bee is capable of producing its novel endo-beta-1,4 glucanase. Further it could be helpful, in understanding, the carbohydrate metabolism in A mellifera.

Keywords: honey bees, Endo-beta 1, 4- glucanase, Apis mellifera, functional expression

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69 Proteomic Analysis of Excretory Secretory Antigen (ESA) from Entamoeba histolytica HM1: IMSS

Authors: N. Othman, J. Ujang, M. N. Ismail, R. Noordin, B. H. Lim

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Amoebiasis is caused by the Entamoeba histolytica and still endemic in many parts of the tropical region, worldwide. Currently, there is no available vaccine against amoebiasis. Hence, there is an urgent need to develop a vaccine. The excretory secretory antigen (ESA) of E. histolytica is a suitable biomarker for the vaccine candidate since it can modulate the host immune response. Hence, the objective of this study is to identify the proteome of the ESA towards finding suitable biomarker for the vaccine candidate. The non-gel based and gel-based proteomics analyses were performed to identify proteins. Two kinds of mass spectrometry with different ionization systems were utilized i.e. LC-MS/MS (ESI) and MALDI-TOF/TOF. Then, the functional proteins classification analysis was performed using PANTHER software. Combination of the LC -MS/MS for the non-gel based and MALDI-TOF/TOF for the gel-based approaches identified a total of 273 proteins from the ESA. Both systems identified 29 similar proteins whereby 239 and 5 more proteins were identified by LC-MS/MS and MALDI-TOF/TOF, respectively. Functional classification analysis showed the majority of proteins involved in the metabolic process (24%), primary metabolic process (19%) and protein metabolic process (10%). Thus, this study has revealed the proteome the E. histolytica ESA and the identified proteins merit further investigations as a vaccine candidate.

Keywords: E. histolytica, ESA, proteomics, biomarker

Procedia PDF Downloads 307
68 Combined Proteomic and Metabolomic Analysis Approaches to Investigate the Modification in the Proteome and Metabolome of in vitro Models Treated with Gold Nanoparticles (AuNPs)

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

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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

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67 High Throughput LC-MS/MS Studies on Sperm Proteome of Malnad Gidda (Bos Indicus) Cattle

Authors: Kerekoppa Puttaiah Bhatta Ramesha, Uday Kannegundla, Praseeda Mol, Lathika Gopalakrishnan, Jagish Kour Reen, Gourav Dey, Manish Kumar, Sakthivel Jeyakumar, Arumugam Kumaresan, Kiran Kumar M., Thottethodi Subrahmanya Keshava Prasad

Abstract:

Spermatozoa are the highly specialized transcriptionally and translationally inactive haploid male gamete. The understanding of proteome of sperm is indispensable to explore the mechanism of sperm motility and fertility. Though there is a large number of human sperm proteomic studies, in-depth proteomic information on Bos indicus spermatozoa is not well established yet. Therefore, we illustrated the profile of sperm proteome in indigenous cattle, Malnad gidda (Bos Indicus), using high-resolution mass spectrometry. In the current study, two semen ejaculates from 3 breeding bulls were collected employing the artificial vaginal method. Using 45% percoll purification, spermatozoa cells were isolated. Protein was extracted using lysis buffer containing 2% Sodium Dodecyl Sulphate (SDS) and protein concentration was estimated. Fifty micrograms of protein from each individual were pooled for further downstream processing. Pooled sample was fractionated using SDS-Poly Acrylamide Gel Electrophoresis, which is followed by in-gel digestion. The peptides were subjected to C18 Stage Tip clean-up and analyzed in Orbitrap Fusion Tribrid mass spectrometer interfaced with Proxeon Easy-nano LC II system (Thermo Scientific, Bremen, Germany). We identified a total of 6773 peptides with 28426 peptide spectral matches, which belonged to 1081 proteins. Gene ontology analysis has been carried out to determine the biological processes, molecular functions and cellular components associated with sperm protein. The biological process chiefly represented our data is an oxidation-reduction process (5%), spermatogenesis (2.5%) and spermatid development (1.4%). The highlighted molecular functions are ATP, and GTP binding (14%) and the prominent cellular components most observed in our data were nuclear membrane (1.5%), acrosomal vesicle (1.4%), and motile cilium (1.3%). Seventeen percent of sperm proteins identified in this study were involved in metabolic pathways. To the best of our knowledge, this data represents the first total sperm proteome from indigenous cattle, Malnad Gidda. We believe that our preliminary findings could provide a strong base for the future understanding of bovine sperm proteomics.

Keywords: Bos indicus, Malnad Gidda, mass spectrometry, spermatozoa

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66 Proteomic Analysis of the Inhibition of Prolyl Oligopeptidase Induced by Z-Pro-Prolinal in Filarial Parasites

Authors: Mohit Wadhawan, Sushma Rathaur

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Lymphatic filariasis, also called elephantiasis is a tropical disease afflicting over 120 million people in 81 countries worldwide. Existing anti filarial drugs are effective against the larval stages of filarial parasites which call for an urgent need of drugs which are macrofilaricidal. Identification of molecular targets crucial for survival of filarial parasites is a prerequisite for drug designing. Prolyl oligopeptidase (POP) is one such crucial enzyme involved in the maturation and degradation of neuropeptides and peptide hormones. We have identified this peptidase in the bovine filarial parasite, Setaria cervi. Effect of inhibition of POP on the proteome profile of filarial parasite has been discussed in this study. Filarial parasites were exposed to Z-pro-prolinal (ZPP), a specific POP inhibitor for 8 h and the motility and viability of the parasites was observed. It significantly reduced the motility and viability of the parasites. To study the proteome profile, the cytosolic, endoplasmic reticulum (ER) and mitochondrial extracts of the adult female parasites were subjected to 2-dimensional electrophoresis. As analyzed by the PD-Quest software, the ZPP caused the alteration in the different subcellular proteins, and the significantly altered proteins were identified using MALDI-MS/MS spectrometry. The major proteins identified were found to play important role in diverse biological functions like signaling, redox regulation, energy metabolism, stress response, and cytoskeleton formation. Moreover, we found upregulation in the calcium binding proteins such as calreticulin, calponin, and calpain-6 suggesting that POP inhibition regulates calcium release. This relates to earlier reports that POP plays non-catalytic role in inositol 1,4,5-trisphosphate (IP3) signaling inducing release of calcium from ER. Taken together, the data demonstrated that inhibition of prolyl oligopeptidase alter the overall proteome signifying its role in survival of the filarial parasites. Thus this study provides a basis for the use of POP as a chemotherapeutic target for the treatment of lymphatic filariasis.

Keywords: lymphatic filariasis, setaria cervi, prolyl oligopeptidase, proteomics

Procedia PDF Downloads 253
65 A Laser Instrument Rapid-E+ for Real-Time Measurements of Airborne Bioaerosols Such as Bacteria, Fungi, and Pollen

Authors: Minghui Zhang, Sirine Fkaier, Sabri Fernana, Svetlana Kiseleva, Denis Kiselev

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The real-time identification of bacteria and fungi is difficult because they emit much weaker signals than pollen. In 2020, Plair developed Rapid-E+, which extends abilities of Rapid-E to detect smaller bioaerosols such as bacteria and fungal spores with diameters down to 0.3 µm, while keeping the similar or even better capability for measurements of large bioaerosols like pollen. Rapid-E+ enables simultaneous measurements of (1) time-resolved, polarization and angle dependent Mie scattering patterns, (2) fluorescence spectra resolved in 16 channels, and (3) fluorescence lifetime of individual particles. Moreover, (4) it provides 2D Mie scattering images which give the full information on particle morphology. The parameters of every single bioaerosol aspired into the instrument are subsequently analysed by machine learning. Firstly, pure species of microbes, e.g., Bacillus subtilis (a species of bacteria), and Penicillium chrysogenum (a species of fungal spores), were aerosolized in a bioaerosol chamber for Rapid-E+ training. Afterwards, we tested microbes under different concentrations. We used several steps of data analysis to classify and identify microbes. All single particles were analysed by the parameters of light scattering and fluorescence in the following steps. (1) They were treated with a smart filter block to get rid of non-microbes. (2) By classification algorithm, we verified the filtered particles were microbes based on the calibration data. (3) The probability threshold (defined by the user) step provides the probability of being microbes ranging from 0 to 100%. We demonstrate how Rapid-E+ identified simultaneously microbes based on the results of Bacillus subtilis (bacteria) and Penicillium chrysogenum (fungal spores). By using machine learning, Rapid-E+ achieved identification precision of 99% against the background. The further classification suggests the precision of 87% and 89% for Bacillus subtilis and Penicillium chrysogenum, respectively. The developed algorithm was subsequently used to evaluate the performance of microbe classification and quantification in real-time. The bacteria and fungi were aerosolized again in the chamber with different concentrations. Rapid-E+ can classify different types of microbes and then quantify them in real-time. Rapid-E+ enables classifying different types of microbes and quantifying them in real-time. Rapid-E+ can identify pollen down to species with similar or even better performance than the previous version (Rapid-E). Therefore, Rapid-E+ is an all-in-one instrument which classifies and quantifies not only pollen, but also bacteria and fungi. Based on the machine learning platform, the user can further develop proprietary algorithms for specific microbes (e.g., virus aerosols) and other aerosols (e.g., combustion-related particles that contain polycyclic aromatic hydrocarbons).

Keywords: bioaerosols, laser-induced fluorescence, Mie-scattering, microorganisms

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64 Identification and Characterization of Small Peptides Encoded by Small Open Reading Frames using Mass Spectrometry and Bioinformatics

Authors: Su Mon Saw, Joe Rothnagel

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Short open reading frames (sORFs) located in 5’UTR of mRNAs are known as uORFs. Characterization of uORF-encoded peptides (uPEPs) i.e., a subset of short open reading frame encoded peptides (sPEPs) and their translation regulation lead to understanding of causes of genetic disease, proteome complexity and development of treatments. Existence of uORFs within cellular proteome could be detected by LC-MS/MS. The ability of uORF to be translated into uPEP and achievement of uPEP identification will allow uPEP’s characterization, structures, functions, subcellular localization, evolutionary maintenance (conservation in human and other species) and abundance in cells. It is hypothesized that a subset of sORFs are translatable and that their encoded sPEPs are functional and are endogenously expressed contributing to the eukaryotic cellular proteome complexity. This project aimed to investigate whether sORFs encode functional peptides. Liquid chromatography-mass spectrometry (LC-MS) and bioinformatics were thus employed. Due to probable low abundance of sPEPs and small in sizes, the need for efficient peptide enrichment strategies for enriching small proteins and depleting the sub-proteome of large and abundant proteins is crucial for identifying sPEPs. Low molecular weight proteins were extracted using SDS-PAGE from Human Embryonic Kidney (HEK293) cells and Strong Cation Exchange Chromatography (SCX) from secreted HEK293 cells. Extracted proteins were digested by trypsin to peptides, which were detected by LC-MS/MS. The MS/MS data obtained was searched against Swiss-Prot using MASCOT version 2.4 to filter out known proteins, and all unmatched spectra were re-searched against human RefSeq database. ProteinPilot v5.0.1 was used to identify sPEPs by searching against human RefSeq, Vanderperre and Human Alternative Open Reading Frame (HaltORF) databases. Potential sPEPs were analyzed by bioinformatics. Since SDS PAGE electrophoresis could not separate proteins <20kDa, this could not identify sPEPs. All MASCOT-identified peptide fragments were parts of main open reading frame (mORF) by ORF Finder search and blastp search. No sPEP was detected and existence of sPEPs could not be identified in this study. 13 translated sORFs in HEK293 cells by mass spectrometry in previous studies were characterized by bioinformatics. Identified sPEPs from previous studies were <100 amino acids and <15 kDa. Bioinformatics results showed that sORFs are translated to sPEPs and contribute to proteome complexity. uPEP translated from uORF of SLC35A4 was strongly conserved in human and mouse while uPEP translated from uORF of MKKS was strongly conserved in human and Rhesus monkey. Cross-species conserved uORFs in association with protein translation strongly suggest evolutionary maintenance of coding sequence and indicate probable functional expression of peptides encoded within these uORFs. Translation of sORFs was confirmed by mass spectrometry and sPEPs were characterized with bioinformatics.

Keywords: bioinformatics, HEK293 cells, liquid chromatography-mass spectrometry, ProteinPilot, Strong Cation Exchange Chromatography, SDS-PAGE, sPEPs

Procedia PDF Downloads 157
63 Nectariferous Plant Genetic Resources for Apicultural Entrepreneurship in Nigeria: Prerequisite for Conservation, Sustainable Management and Policy

Authors: C. V. Nnamani, O. L. Adedeji

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The contemporary global economic meltdown has devastating effect on the Nigerian’s economy and its frantic search for alternative source of national revenue aside from oil and gas has become imperative for economic emancipation for Nigerians. Apicultural entrepreneurship could provide a source of livelihood if the basic knowledge of those plant genetic resources needed by bees is made available. A palynological evaluation of those palynotaxa which honey bees forage for pollen and nectar was carried out after standard acetolysis method. Results showed that the honey samples were highly diversified and rich in honey plants. A total of 9544.3 honey pollen, consisting of 39 honey plants belonging to 21 plant families and distributed within 38 genera were identified excluding 238 unidentified pollen grains. Data from the analysis equally revealed that Elaeis guineensis Jacq, Anacardium occidentale L, Diospyros mespiliformis Hochist xe ADC, Alchornea cordifolia Muell, Arg, Daniella oliveri (Rolfe) Hutch & Dalz, Irvingia wombolu Okafor ex Baill, Treculia africana Decne, Nauclea latifolia Smith and Crossopteryx febrifuga Afzil ex Benth were the predominant honey plants. It provided a guide to the optimal utilization of floral resources by honeybees in these regions, showing the opportunity and amazing potentials for apiculture entrepreneurship of these palytaxa. Most of these plants are rare, threatened and endangered. It calls for urgent conservation techniques and step by all players. Critical awareness creation to ensure farmers knowledge of these palynotaxa to ensure proper understanding and attendance boost from them as economic empowerment is needed.

Keywords: palynotaxa, acetolysis, enterprise, livelihood, Nigeria

Procedia PDF Downloads 264
62 Green Synthesis of Silver Nanoparticles from Citrus aurantium Aqueous Pollen Extract and Their Antibacterial Activity

Authors: Mohammad Ali Karimi, Hossein Tavallali, Abdolhamid Hatefi-Mehrjardi

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Pollen extract of in vitro plants raised of Citrus aurantium as reducer and stabilizer was assessed for the green synthesis of silver nanoparticles (AgNPs). The synthesis of AgNPs was performed at room temperature assisting in solutions by reduction takes place rapidly for 10 min. Surface plasmon resonance (SPR) peaks in UV–Vis spectra indicated the formation of polydispersive AgNPs. Silver ions concentration, pH, temperature and reaction time were optimized in the synthesis of AgNPs. The nanoparticles obtained were characterized by UV-Vis spectrophotometer, transmission electron microscopy (TEM). X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy techniques. The synthesized AgNPs were mostly spherical in shape with an average size of 15 nm. XRD study shows that the AgNPs are crystalline in nature with face-centered cubic (fcc) geometry. It shows the significant antibacterial efficacy against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) by disk diffusion method using Mueller-Hinton Agar.

Keywords: green synthesis, Citrus aurantium, silver nanoparticles, antibacterial activity

Procedia PDF Downloads 258
61 Segmentation Using Multi-Thresholded Sobel Images: Application to the Separation of Stuck Pollen Grains

Authors: Endrick Barnacin, Jean-Luc Henry, Jimmy Nagau, Jack Molinie

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Being able to identify biological particles such as spores, viruses, or pollens is important for health care professionals, as it allows for appropriate therapeutic management of patients. Optical microscopy is a technology widely used for the analysis of these types of microorganisms, because, compared to other types of microscopy, it is not expensive. The analysis of an optical microscope slide is a tedious and time-consuming task when done manually. However, using machine learning and computer vision, this process can be automated. The first step of an automated microscope slide image analysis process is segmentation. During this step, the biological particles are localized and extracted. Very often, the use of an automatic thresholding method is sufficient to locate and extract the particles. However, in some cases, the particles are not extracted individually because they are stuck to other biological elements. In this paper, we propose a stuck particles separation method based on the use of the Sobel operator and thresholding. We illustrate it by applying it to the separation of 813 images of adjacent pollen grains. The method correctly separated 95.4% of these images.

Keywords: image segmentation, stuck particles separation, Sobel operator, thresholding

Procedia PDF Downloads 101