Search results for: enzootic bovine leukosis

Authors: Eman M. Sarhan, Doaa A. Ghareeb, Gabriella Ortore, Amr A. Amara, Mohamed M. El-Sayed

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Drug salting and nanoparticle-based drug delivery formulations are considered to be an effective means for rendering the hydrophobic drugs’ nano-scale dispersion in aqueous media, and thus circumventing the pitfalls of their poor solubility as well as enhancing their membrane permeability. The current study aims to increase the bioavailability of quaternary ammonium berberine through acid salting and biodegradable bovine serum albumin (BSA)-based nanoparticulate drug formulation. Berberine hydroxide (BBR-OH) that was chemically synthesized by alkalization of the commercially available berberine hydrochloride (BBR-HCl) was then acidified to get Di-berberine sulfate (BBR)₂SO₄. The purified crystals were spectrally characterized. The desolvation technique was optimized for the preparation of size-controlled BSA-BBR-HCl, BSA-BBR-OH, and BSA-(BBR)₂SO₄ nanoparticles. Particle size, zeta potential, drug release, encapsulation efficiency, Fourier transform infrared spectroscopy (FTIR), tandem MS-MS spectroscopy, energy-dispersive X-ray spectroscopy (EDX), scanning and transmitting electron microscopic examination (SEM, TEM), in vitro bioactivity, and in silico drug-polymer interaction were determined. BSA (PDB ID; 4OR0) protonation state at different pH values was predicted using Amber12 molecular dynamic simulation. Then blind docking was performed using Lamarkian genetic algorithm (LGA) through AutoDock4.2 software. Results proved the purity and the size-controlled synthesis of berberine-BSA-nanoparticles. The possible binding poses, hydrophobic and hydrophilic interactions of berberine on BSA at different pH values were predicted. Antioxidant, anti-hemolytic, and cell differentiated ability of tested drugs and their nano-formulations were evaluated. Thus, drug salting and the potentially effective albumin berberine nanoparticle formulations can be successfully developed using a well-optimized desolvation technique and exhibiting better in vitro cellular bioavailability.

Keywords: berberine, BSA, BBR-OH, BBR-HCl, BSA-BBR-HCl, BSA-BBR-OH, (BBR)₂SO₄, BSA-(BBR)₂SO₄, FTIR, AutoDock4.2 Software, Lamarkian genetic algorithm, SEM, TEM, EDX

Procedia PDF Downloads 174

Authors: Jovana Bogojeski, Dusan Cocic, Nenad Jankovic, Angelina Petrovic

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Kinetically-inert transition metal complexes, such as Rh(III) and Os(III) complexes, attract increasing attention as leading scaffolds for the development of potential pharmacological agents due to their inertness and stability. Therefore, we have designed and fully characterized a few novel rhodium(III) and osmium(III) complexes with a tridentate nitrogen−donor chelate system. For some complexes, the crystal X-ray structure analysis was performed. Reactivity of the newly synthesized complexes towards small biomolecules, such as L-methionine (L-Met), guanosine-5’-monophosphate (5’-GMP), and glutathione (GSH) has been examined. Also, the reactivity of these complexes towards the DNA/RNA (Ribonucleic acid) duplexes was investigated. Obtained results show that the newly synthesized complexes exhibit good affinity towards the studied ligands. Results also show that the complexes react faster with the RNA duplex than with the DNA and that in the DNA duplex reaction is faster with 15mer GG than with the 22mer GG. The UV-Vis (Ultraviolet-visible spectroscopy) is absorption spectroscopy, and the EB (Ethidium bromide) displacement studies were used to examine the interaction of these complexes with CT-DNA and BSA (Bovine serum albumin). All studied complex showed good interaction ability with both the DNA and BSA. Furthermore, the DFT (Density-functional theory) calculation and docking studies were performed. The impact of the metal complex on the cytotoxicity was tested by MTT assay (a colorimetric assay for assessing cell metabolic activity) on HCT-116 lines (human colon cancer cell line). In addition, all these tests were repeated in the presence of several water-soluble biologically active ionic liquids. Attained results indicate that the ionic liquids increase the activity of the investigated complexes. All obtained results in this study imply that the introduction of different spectator ligand can be used to improve the reactivity of rhodium(III) and osmium(III) complexes. Finally, these results indicate that the examined complexes show reactivity characteristics needed for potential anti-tumor agents, with possible targets being both the DNA and proteins. Every new contribution in this field is highly warranted due to the current lack of clinically used Metallo-based alternatives to cisplatin.

Keywords: biomolecules, ionic liquids, osmium(III), rhodium(III)

Procedia PDF Downloads 150

Authors: Yasser Y. Lenis, Amy Jo Montgomery, Diego F. Carrillo-Gonzalez

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Introduction: The use of didactic instruments in different learning environments plays a pivotal role in enhancing the level of knowledge in veterinary science students. The direct instruction of basic animal reproduction concepts in students enrolled in veterinary medicine programs allows them to elucidate the biological and molecular mechanisms that perpetuate the animal species in an ecosystem. Therefore, universities must implement didactic strategies that facilitate the teaching and learning processes for students and, in turn, enrich learning environments. Objective: to evaluate the effect of the use of a didactic textbook on the level of theoretical knowledge in embryo-maternal recognition for veterinary medicine students. Methods: the participants (n=24) were divided into two experimental groups: control (Ctrl) and treatment (Treat). Both groups received 4 hours of theoretical training regarding the basic concepts in bovine embryo-maternal recognition. However, the Treat group was also exposed to a guided lecture and the activity play-to-learn from a cow reproduction didactic textbook. A pre-test and a post-test were applied to assess the prior and subsequent knowledge in the participants. Descriptive statistics were applied to identify the success rates for each of the tests. Afterwards, a repeated measures model was applied where the effect of the intervention was considered. Results: no significant difference (p>0,05) was observed in the number of right answers for groups Ctrl (54,2%±12,7) and Treat (40,8%±16,8) in the pre-test. There was no difference (p>0,05) compering the number of right answers in Ctrl pre-test (54,2%±12,7) and post-test (60,8±18,8). However, the Treat group showed a significant (p>0,05) difference in the number of right answers when comparing pre-test (40,8%±16,8) and post-test (71,7%±14,7). Finally, after the theoretical training and the didactic activity in the Treat group, an increase of 10.9% (p<0,05) in the number of right answers was found when compared with the Ctrl group. Conclusion: the use of didactic tools that include guided lectures and activities like play-to-learn from a didactic textbook enhances the level of knowledge in an animal reproduction course for veterinary medicine students.

Keywords: animal reproduction, pedagogic, level of knowledge, learning environment

Procedia PDF Downloads 65

Authors: Z. Arabshahi, I. Afara, A. Oloyede, H. Moody, J. Kashani, T. Klein

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Articular cartilage is a fluid-swollen tissue of synovial joints that functions by providing a lubricated surface for articulation and to facilitate the load transmission. The biomechanical function of this tissue is highly dependent on the integrity of its ultrastructural matrix. Any alteration of articular cartilage matrix, either by injury or degenerative conditions such as osteoarthritis (OA), compromises its functional behaviour. Therefore, the assessment of articular cartilage is important in early stages of degenerative process to prevent or reduce further joint damage with associated socio-economic impact. Therefore, there has been increasing research interest into the functional assessment of articular cartilage. This study developed a characterization parameter for articular cartilage assessment based on the response of cartilage surface to loading/unloading. This is because the response of articular cartilage to compressive loading is significantly depth-dependent, where the superficial zone and underlying matrix respond differently to deformation. In addition, the alteration of cartilage matrix in the early stages of degeneration is often characterized by PG loss in the superficial layer. In this study, it is hypothesized that the response of superficial layer is different in normal and proteoglycan depleted tissue. To establish the viability of this hypothesis, samples of visually intact and artificially proteoglycan-depleted bovine cartilage were subjected to compression at a constant rate to 30 percent strain using a ring-shaped indenter with an integrated ultrasound probe and then unloaded. The response of articular surface which was indirectly loaded was monitored using ultrasound during the time of loading/unloading (deformation/recovery). It was observed that the rate of cartilage surface response to loading/unloading was different for normal and PG-depleted cartilage samples. Principal Component Analysis was performed to identify the capability of the cartilage surface response to loading/unloading, to distinguish between normal and artificially degenerated cartilage samples. The classification analysis of this parameter showed an overlap between normal and degenerated samples during loading. While there was a clear distinction between normal and degenerated samples during unloading. This study showed that the cartilage surface response to loading/unloading has the potential to be used as a parameter for cartilage assessment.

Keywords: cartilage integrity parameter, cartilage deformation/recovery, cartilage functional assessment, ultrasound

Procedia PDF Downloads 192

Authors: Norhidayu Muhamad Zain

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Gelatin, a purified protein derived mostly from porcine and bovine sources, is used widely in food manufacturing, pharmaceutical, and cosmetic industries. However, the presence of any porcine-related products are strictly forbidden for Muslim and Jewish consumption. Therefore, analytical methods offering reliable results to differentiate the sources of gelatin are needed. The aim of this study was to differentiate the sources of gelatin (porcine and tilapia fish) using Fourier transform infrared- attenuated total reflection (FTIR-ATR) combined with two dimensional infrared (2DIR) correlation analysis. Porcine gelatin (PG) and tilapia fish gelatin (FG) samples were diluted in distilled water at concentrations ranged from 4-20% (w/v). The samples were then analysed using FTIR-ATR and 2DIR correlation software. The results showed a significant difference in the pattern map of synchronous spectra at the region of 1000 cm⁻¹ to 1100 cm⁻¹ between PG and FG samples. The auto peak at 1080 cm⁻¹ that attributed to C-O functional group was observed at high intensity in PG samples compared to FG samples. Meanwhile, two auto peaks (1080 cm⁻¹ and 1030 cm⁻¹) at lower intensity were identified in FG samples. In addition, using 2D correlation analysis, the original broad water OH bands in 1D IR spectra can be effectively differentiated into six auto peaks located at 3630, 3340, 3230, 3065, 2950 and 2885 cm⁻¹ for PG samples and five auto peaks at 3630, 3330, 3230, 3060 and 2940 cm⁻¹ for FG samples. Based on the rule proposed by Noda, the sequence of the spectral changes in PG samples is as following: NH₃⁺ amino acid > CH₂ and CH₃ aliphatic > OH stretch > carboxylic acid OH stretch > NH in secondary amide > NH in primary amide. In contrast, the sequence was totally in the opposite direction for FG samples and thus both samples provide different 2D correlation spectra ranged from 2800 cm-1 to 3700 cm⁻¹. This method may provide a rapid determination of gelatin source for application in food, pharmaceutical, and cosmetic products.

Keywords: 2 dimensional infrared (2DIR) correlation analysis, Fourier transform infrared- attenuated total reflection (FTIR-ATR), porcine gelatin, tilapia fish gelatin

Procedia PDF Downloads 250

Authors: Rima Zakzouk, Yasushi Shimada, Yuan Zhou, Yasunori Sumi, Junji Tagami

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Background and Purpose: Swept source optical coherence tomography (OCT) is an interferometric imaging technique that has been recently used in cariology. In spite of progress made in adhesive dentistry, the composite restoration has been failing due to secondary caries which occur due to environmental factors in oral cavities. Therefore, a precise assessment to effective marginal sealing of restoration is highly required. The aim of this study was evaluating gap formation at composite/cavity walls interface with or without phosphoric acid etching using SS-OCT. Materials and Methods: Round tapered cavities (2×2 mm) were prepared in three locations, mid-coronal, cervical, and root of bovine incisors teeth in two groups (SE and PA Groups). While self-etching adhesive (Clearfil SE Bond) was applied for the both groups, Group PA had been already pretreated with phosphoric acid etching (K-Etchant gel). Subsequently, both groups were restored by Estelite Flow Quick Flowable Composite Resin. Following 5000 thermal cycles, three cross-sectionals were obtained from each cavity using OCT at 1310-nm wavelength at 0°, 60°, 120° degrees. Scanning was repeated after two months to monitor the gap progress. Then the average percentage of gap length was calculated using image analysis software, and the difference of mean between both groups was statistically analyzed by t-test. Subsequently, the results were confirmed by sectioning and observing representative specimens under Confocal Laser Scanning Microscope (CLSM). Results: The results showed that pretreatment with phosphoric acid etching, Group PA, led to significantly bigger gaps in mid-coronal and cervical compared to SE group, while in the root cavity no significant difference was observed between both groups. On the other hand, the gaps formed in root’s cavities were significantly bigger than those in mid-coronal and cervical within the same group. This study investigated the effect of phosphoric acid on gap length progress on the composite restorations. In conclusions, phosphoric acid etching treatment did not reduce the gap formation even in different regions of the tooth. Significance: The cervical region of tooth was more exposing to gap formation than mid-coronal region, especially when we added pre-etching treatment.

Keywords: image analysis, optical coherence tomography, phosphoric acid etching, self-etch adhesives

Procedia PDF Downloads 221

Authors: Himani Sharma, Amit Agrawal

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Stability of the Cassie and Wenzel wetting states depends on intrinsic contact angle and geometric features on a surface that was exploited in capturing biofluids in microwells. However, the mechanism of imbibition of biofluids in the microwells is not well implied in terms of wettability of a substrate. In this work, we experimentally demonstrated filling dynamics in hydrophilic and hydrophobic microwells by protein solutions. Towards this, we utilized lotus leaf as a mold to fabricate microwells on a Polydimethylsiloxane (PDMS) surface. Lotus leaf containing micrometer-sized blunt-conical shaped pillars with a height of 8-15 µm and diameter of 3-8 µm were transferred on to PDMS. Furthermore, PDMS surface was treated with oxygen plasma to render the hydrophilic nature. A 10µL droplets containing fluorescein isothiocyanate (FITC) - labelled bovine serum albumin (BSA) were rested on both hydrophobic (θa = 108o, where θa is the apparent contact angle) and hydrophilic (θa = 60o) PDMS surfaces. A time-dependent fluorescence microscopy was conducted on these modified PDMS surfaces by recording the fluorescent intensity over a 5 minute period. It was observed that, initially (at t=1 min) FITC-BSA was accumulated on the periphery of both hydrophilic and hydrophobic microwells due to incomplete penetration of liquid-gas meniscus. This deposition of FITC-BSA on periphery of microwell was not changed with time for hydrophobic surfaces, whereas, a complete filling was occurred in hydrophilic microwells (at t=5 mins). This attributes to a gradual movement of three-phase contact line along the vertical surface of the hydrophilic microwells as compared to stable pinning in the hydrophobic microwells as confirmed by Surface Evolver simulations. In addition, if the cavities are presented on hydrophobic surfaces, air bubbles will be trapped inside the cavities once the aqueous solution is placed over these surfaces, resulting in the Cassie-Baxter wetting state. This condition hinders trapping of proteins inside the microwells. Thus, it is necessary to impart hydrophilicity to the microwell surfaces so as to induce the Wenzel state, such that, an entire solution will be fully in contact with the walls of microwells. Imbibition of microwells by protein solutions was analyzed in terms fluorescent intensity versus time. The present work underlines the importance of geometry of microwells and surface wettability of substrate in wetting and effective capturing of solid sub-phases in biofluids.

Keywords: BSA, microwells, surface evolver, wettability

Procedia PDF Downloads 198

Authors: Zhen Cao, Yu Zhu, Junxue Fu

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Immunoassay, i.e., antigen-antibody reaction, is crucial for disease diagnostics. To achieve the adequate signal of the antigen protein detection, a large amount of sample and long incubation time is needed. However, the amount of protein is usually small at the early stage, which makes it difficult to detect. Unlike cells and DNAs, no valid chemical method exists for protein amplification. Thus, an alternative way to improve the signal is through particle manipulation techniques to concentrate proteins, among which dielectrophoresis (DEP) is an effective one. DEP is a technique that concentrates particles to the designated region through a force created by the gradient in a non-uniform electric field. Since DEP force is proportional to the cube of particle size and square of electric field gradient, it is relatively easy to capture larger particles such as cells. For smaller ones like proteins, a super high gradient is then required. In this work, three-dimensional Ag/SiO2 nanorods arrays, fabricated by an easy physical vapor deposition technique called as oblique angle deposition, have been integrated with a DEP device and created the field gradient as high as of 2.6×10²⁴ V²/m³. The nanorods based DEP device is able to enrich bovine serum albumin (BSA) protein by 1800-fold and the rate has reached 180-fold/s when only applying 5 V electric potential. Based on the above nanorods integrated DEP platform, an immunoassay of mouse immunoglobulin G (IgG) proteins has been performed. Briefly, specific antibodies are immobilized onto nanorods, then IgG proteins are concentrated and captured, and finally, the signal from fluorescence-labelled antibodies are detected. The limit of detection (LoD) is measured as 275.3 fg/mL (~1.8 fM), which is a 20,000-fold enhancement compared with identical assays performed on blank glass plates. Further, prostate-specific antigen (PSA), which is a cancer biomarker for diagnosis of prostate cancer after radical prostatectomy, is also quantified with a LoD as low as 2.6 pg/mL. The time to signal saturation has been significantly reduced to one minute. In summary, together with an easy nanorod fabrication and integration method, this nanorods based DEP platform has demonstrated highly sensitive immunoassay performance and thus poses great potentials in applications for early point-of-care diagnostics.

Keywords: dielectrophoresis, immunoassay, oblique angle deposition, protein concentration

Procedia PDF Downloads 103

Authors: Abdulqader Alkhouzaam, Hazim Qiblawey, Majeda Khraisheh

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Membrane treatment for desalination and wastewater treatment is one of the promising solutions to affordable clean water. It is a developing technology throughout the world and considered as the most effective and economical method available. However, the limitations of membranes’ mechanical and chemical properties restrict their industrial applications. Hence, developing novel membranes was the focus of most studies in the water treatment and desalination sector to find new materials that can improve the separation efficiency while reducing membrane fouling, which is the most important challenge in this field. Graphene oxide (GO) is one of the materials that have been recently investigated in the membrane water treatment sector. In this work, ultrafiltration polysulfone (PSF) membranes with high antifouling properties were synthesized by incorporating different loadings of GO. High-oxidation degree GO had been synthesized using a modified Hummers' method. The synthesized GO was characterized using different analytical techniques including elemental analysis, Fourier transform infrared spectroscopy - universal attenuated total reflectance sensor (FTIR-UATR), Raman spectroscopy, and CHNSO elemental analysis. CHNSO analysis showed a high oxidation degree of GO represented by its oxygen content (50 wt.%). Then, ultrafiltration PSF membranes incorporating GO were fabricated using the phase inversion technique. The prepared membranes were characterized using scanning electron microscopy (SEM) and atomic force microscopy (AFM) and showed a clear effect of GO on PSF physical structure and morphology. The water contact angle of the membranes was measured and showed better hydrophilicity of GO membranes compared to pure PSF caused by the hydrophilic nature of GO. Separation properties of the prepared membranes were investigated using a cross-flow membrane system. Antifouling properties were studied using bovine serum albumin (BSA) and humic acid (HA) as model foulants. It has been found that GO-based membranes exhibit higher antifouling properties compared to pure PSF. When using BSA, the flux recovery ratio (FRR %) increased from 65.4 ± 0.9 % for pure PSF to 84.0 ± 1.0 % with a loading of 0.05 wt.% GO in PSF. When using HA as model foulant, FRR increased from 87.8 ± 0.6 % to 93.1 ± 1.1 % with 0.02 wt.% of GO in PSF. The pure water permeability (PWP) decreased with loadings of GO from 181.7 L.m⁻².h⁻¹.bar⁻¹ of pure PSF to 181.1, and 157.6 L.m⁻².h⁻¹.bar⁻¹ with 0.02 and 0.05 wt.% GO respectively. It can be concluded from the obtained results that incorporating low loading of GO could enhance the antifouling properties of PSF hence improving its lifetime and reuse.

Keywords: antifouling properties, GO based membranes, hydrophilicity, polysulfone, ultrafiltration

Procedia PDF Downloads 143

Authors: Brian Chi Yan Cheng, Hui Guo, Tao Su, Xiu‐qiong Fu, Ting Li, Zhi‐ling Yu

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Rheumatoid arthritis (RA) affects around 1% of the globe population. Yet, there is still no cure for RA. Toll-like receptor 4 (TLR4) signalling has been found to be involved in the pathogenesis of RA, making it a potential therapeutic target for RA treatment. A herbal formula (RL) consisting of fruits of Rosa Multiflora (Eijitsu rose) and flowers of Lonicera Japonica (Japanese honeysuckle) has been used in treating various inflammatory disorders for more than a thousand year. Both of them are rich sources of nutrients and bioactive phytochemicals, which can be used in producing different food products and supplements. In this study, we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis (CIA) in rats and investigate the involvement of TLR4 signaling in the mode of action of RL. Anti-arthritic efficacy was evaluated using CIA rats induced by bovine type II collagen. The treatment groups were treated with RL (82.5, 165, and 330 mg/kg bw per day, p.o.) or positive control indomethacin (0.25 mg/kg bw per day, p.o.) for 35 days. Clinical signs (hind paw volume and arthritis severity scores), changes in serum inflammatory mediators, pro-/antioxidant status, histological and radiographic changes of joints were investigated. Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats. The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats, murine RAW264.7 macrophages and human THP-1 monocytic cells. The severity of arthritis in the CIA rats was significantly attenuated by RL. Antioxidant status, histological score and radiographic score were efficiently improved by RL. RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats. RL significantly inhibited the production of various pro-inflammatory mediators, the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines. RL possesses anti-arthritic effect on collagen-induced arthritis in rats. The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum. The inhibition of the TAK1/NF-κB and TAK1/MAPK pathways participate in the anti-arthritic effects of RL. This provides a pharmacological justification for the dietary use of RL in the control of various arthritic diseases. Further investigation should be done to develop RL into a anti-arthritic food products and/or supplements.

Keywords: japanese honeysuckle, rheumatoid arthritis, rosa multiflora, rosehip

Procedia PDF Downloads 432

Authors: Rima Zakzouk, Yasushi Shimada, Yasunori Sumi, Junji Tagami

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Background and purpose: The resin composite has become the main material for the restorations of caries in recent years due to aesthetic characteristics, especially with the development of the adhesive techniques. The quality of adhesion to tooth structures is depending on an exchange process between inorganic tooth material and synthetic resin and a micromechanical retention promoted by resin infiltration in partially demineralized dentin. Optical coherence tomography (OCT) is a noninvasive diagnostic method for obtaining cross-sectional images that produce high-resolution of the biological tissue at the micron scale. The aim of this study was to evaluate the gap formation at adhesive/tooth interface of two-step self-etch adhesives that are preceded with or without phosphoric acid pre-etching in different regions of teeth using SS-OCT. Materials and methods: Round tapered cavities (2×2 mm) were prepared in cervical part of bovine incisors teeth and divided into 2 groups (n=10): first group self-etch adhesive (Clearfil SE Bond) was applied for SE group and second group treated with acid etching before applying the self-etch adhesive for PA group. Subsequently, both groups were restored with Estelite Flow Quick Flowable Composite Resin and observed under OCT. Following 5000 thermal cycles, the same section was obtained again for each cavity using OCT at 1310-nm wavelength. Scanning was repeated after two months to monitor the gap progress. Then the gap length was measured using image analysis software, and the statistics analysis were done between both groups using SPSS software. After that, the cavities were sectioned and observed under Confocal Laser Scanning Microscope (CLSM) to confirm the result of OCT. Results: Gaps formed at the bottom of the cavity was longer than the gap formed at the margin and dento-enamel junction in both groups. On the other hand, pre-etching treatment led to damage the DEJ regions creating longer gap. After 2 months the results showed almost progress in the gap length significantly at the bottom regions in both groups. In conclusions, phosphoric acid etching treatment did not reduce the gap lrngth in most regions of the cavity. Significance: The bottom region of tooth was more exposed to gap formation than margin and DEJ regions, The DEJ damaged with phosphoric acid treatment.

Keywords: optical coherence tomography, self-etch adhesives, bottom, dento enamel junction

Procedia PDF Downloads 227

Authors: Vanessa Castro, Liria H. Okuda, Daniela P. Chiebao, Adriana H. C. N. Romaldini, Harumi Hojo, Marina Grandi, Joao Paulo A. Silva, Alessandra F. C. Nassar

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The Biological Institute of Sao Paulo, in partnership with a private company, develops an Animal Health Family Farming Program (Prosaf) to enable communication among smallholder farmers and scientists, on-farm consulting and lectures, solving health questions that will benefit agricultural productivity. In Vale do Paraiba region, a dairy region of Sao Paulo State, southern Brazil, many of these types of farms are found with several milk quality problems. Most of these farms are profit-based business; however, with non-technified cattle rearing systems and uncertain veterinary assistance. Feedback from Prosaf showed that the biggest complaints from farmers were low milk production, sick animals and, mainly, loss of selling price due to a high somatic cell count (SCC) and a total bacterial count (TBC). The aims of this study were to improve milk quality, animal hygiene and herd health status by adjustments into general management practices and introducing techniques of sanitary control and milk monitoring in five dairy farms from Sao Jose do Barreiro municipality, Sao Paulo State, Brazil, to increase their profits. A total of 119 milk samples from 56 animals positive for California Mastitis Test (CMT) were collected. The positive CMT indicates subclinical mastitis, therefore laboratorial exams were performed in the milk (microbiological, biochemical and antibiogram test) detect the presence of Staphylococcus aureus (41.8%), Bacillus sp. (11.8%), Streptococcus sp. (2.1%), nonfermenting, motile and oxidase-negative Gram-negative Bacilli (2.1%) and Enterobacter (2.1%). Antibiograms revealed high resistance to gentamicin and streptomycin, probably due to indiscriminate use of antibiotics without veterinarian prescription. We suggested the improvement of hygiene management in the complete milking and cooling tanks system. Using the results of the laboratory tests, animals were properly treated, and the effects observed were better CMT outcomes, lower SCCs, and TBCs leading to an increase in milk pricing. This study will have a positive impact on the family farmers from Sao Paulo State dairy region by improving their market milk competitiveness.

Keywords: milk, family farming, food quality, antibiogram, profitability

Procedia PDF Downloads 155

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

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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

Procedia PDF Downloads 196

Authors: Habtamu Abera Goshu, Ping Yan

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Copy number variation (CNV) is a significant marker of the genetic and phenotypic diversity among individuals that accounts for complex quantitative traits of phenotype and diseases via modulating gene dosage, position effects, alteration of downstream pathways, modification of chromosome structure, and position within the nucleus and disrupting coding regions in the genome. Associating copy number variations (CNVs) with growth and gene expression are a powerful approach for identifying genomic characteristics that contribute to phenotypic and genotypic variation. A previous study using next-generation sequencing illustrated that the choline kinase beta (CHKB), Krüpple-like factor 6 (KLF6), glypican 1(GPC1), and cholinergic receptor muscarinic 3 (CHRM3) genes reside within copy number variable regions (CNVRs) of yak populations that overlap with quantitative trait loci (QTLs) of meat quality and growth. As a result, this research aimed to determine the association of CNVs of the KLF6, CHKB, GPC1, and CHRM3 genes with growth traits in the Datong yak breed. The association between the CNV types of the KLF6, CHKB, GPC1, and CHRM3 genes and the growth traits in the Datong yak breed was determined by one-way analysis of variance (ANOVA) using SPSS software. The CNV types were classified as a loss (a copy number of 0 or 1), gain (a copy number >2), and normal (a copy number of 2) relative to the reference gene, BTF3 in the 387 individuals of Datong yak. These results indicated that the normal CNV types of the CHKB and GPC1 genes were significantly (P<0.05) associated with high body length, height and weight, and chest girth in six-month-old and five-year-old Datong yaks. On the other hand, the loss CNV types of the KLF6 gene is significantly (P<0.05) associated with body weight and length and chest girth at six-month-old and five-year-old Datong yaks. In the contrary, the gain CNV type of the CHRM3 gene is highly (P<0.05) associated with body weight, length, height, and chest girth in six-month-old and five-year-old. This work provides the first observation of the biological role of CNVs of the CHKB, KLF6, GPC1, and CHRM3 genes in the Datong yak breed and might, therefore, provide a novel opportunity to utilize data on CNVs in designing molecular markers for the selection of animal breeding programs for larger populations of various yak breeds. Therefore, we hypothesized that this study provided inclusive information on the application of CNVs of the CHKB, KLF6, GPC1, and CHRM3 genes in growth traits in Datong yaks and its possible function in bovine species.

Keywords: Copy number variation, growth traits, yak, genes

Procedia PDF Downloads 172

Authors: Mohit Kumar, Beklashwar Salona, Shiv Murti, Mukesh Singh

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We report here a case of five-months old male baby, born as second child of non-consanguineous parents with no considerable history of genetic abnormality which was referred to our cytogenetic laboratory for chromosomal analysis. Physical dysmorphic facial features including mongoloid face, cleft palate, simian crease, and developmental delay were observed. We present this case with unique balanced autosomal translocation of t(3;10)(p21;p13). The risk of phenotypic abnormalities based on de novo balanced translocation was estimated to be 7%. The association of balanced chromosomal rearrangement with Down syndrome features such as multiple congenital anomalies, facial dysmorphism and congenital heart anomalies are very rare in a 5-months old male child. Trisomy-21 is not uncommon in chromosomal abnormality with the birth defect and balanced translocations are frequently observed in patients with secondary infertility or recurrent spontaneous abortion (RSA). Two ml heparinized peripheral blood cells cultured in RPMI-1640 for 72 hours supplemented with 20% fetal bovine serum, phytohemagglutinin (PHA), and antibiotics were used for chromosomal analysis. A total 30 metaphases images were captured using Olympus-BX51 microscope and analyzed using Bio-view karyotyping software through GTG-banding (G bands by trypsin and Giemsa) according to International System for Human Cytogenetic Nomenclature 2016. The results showed balanced translocation between short arm of chromosome # 3 and short arm of chromosome # 10. The karyotype of the child was found to be 46,XY,t(3;10)(p21; p13). Chromosomal abnormalities are one of the major causes of birth defect in new born babies. Also, balanced translocations are frequently observed in patients with secondary infertility or recurrent spontaneous abortion. The index case presented with dysmorphic facial features and had a balanced translocation 46,XY,t(3;10)(p21;p13). This translocation with break points at (p21; p13) has not been reported in the literature in a child with facial dysmorphism. To the best of our knowledge, this is the first report of novel balanced translocation t(3;10) with break points in a child with dysmorphic features. We found balanced chromosomal translocation instead of any trisomy or unbalanced aberrations along with some phenotypic abnormalities. Therefore, we suggest that such novel balanced translocation with abnormal phenotype should be reported in order to enable the pathologist, pediatrician, and gynecologist to have a better insight into the intricacies of chromosomal abnormalities and their associated phenotypic features. We hypothesized that dysmorphic features as seen in this case may be the result of change in the pattern of genes located at the breakpoint area in balanced translocations or may be due to deletion or mutation of genes located on the p-arm of chromosome # 3 and p-arm of chromosome # 10.

Keywords: balanced translocation, karyotyping, phenotypic abnormalities, facial dimorphisms

Procedia PDF Downloads 208

Authors: Madelyn N. Moawad, Hermine R. Z. Tadros, Mary G. Ghobrial, Ahmad R. Bassiouny, Kamal M. Kandeel, Athar Ata

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Seaweeds are potential renewable resources in marine environment. They provide an excellent source of bioactive substances such as dietary fibers and various functional polysaccharides that could potentially be used as ingredients for both human and animal health applications. The observations suggested that these bioactive compounds have strong antioxidant properties, which have beneficial effects on human health. The present research aimed at finding new chemical products from local marine macroalgae for natural medicinal uses and consumption for their nutritional values. Macroalgae samples were collected manually mainly from the Mediterranean Sea at shallow subtidal zone of Abu Qir Bay, Alexandria, Egypt. The chemical compositions of lyophilized materials of six selected macroalgal species; Colpomenia sinuosa, Sargassum linifolium, Padina pavonia, Pterocladiella capillacea, Laurencia pinnatifidia, and Caulerpa racemosa, were investigated for proteins using bovine serum albumin, and carbohydrates were assayed by phenol-sulfuric acid reaction. The macroalgae lipid was extracted with chloroform, methanol and phosphate buffer. Vitamins were extracted using trichloroacetic acid. Chlorophylls and total carotenoids were determined spectrophotometrically and total phenols were extracted with methanol. In addition, lipid-soluble, and water-soluble antioxidant, and anti α-glucosidase activities were measured spectrophotometrically. The antioxidant activity of hexane extracts was investigated using phosphomolybdenum reagent. The anti-α-glucosidase effect measurement was initiated by mixing α-glucosidase solution with p-nitrophenyl α-D-glucopyranoside. The results showed that the ash contents varied from 11.2 to 35.4 % on dry weight basis for P. capillacea and Laurencia pinnatifidia, respectively. The protein contents ranged from 5.63 % in brown macroalgae C. sinuosa to 8.73 % in P. pavonia. A relative wide range in carbohydrate contents was observed (20.06–46.75 %) for the test algal species. The highest lipid percentage was found in green alga C. racemosa (5.91%) followed by brown algae P. pavonia (3.57%) and C. sinuosa (2.64%). The phenolic contents varied from 1.32 mg GAE/g for C. sinuosa to 4.00 mg GAE/g in P. pavonia. The lipid-soluble compounds exhibited higher antioxidant capacity (73.18-145.95 µM/g) than that of the water-soluble ones ranging from 24.83 µM/g in C. racemosa to 74.07 µM/g in S. linifolium. The most potent anti-α-glucosidase activity was observed for P. pavonia with IC50 of 17.12 μg/ml followed by S. linifolium (IC50 = 71.75 μg/ml), C. racemosa (IC50 = 84.73 μg/ml), P. capillacea (IC50 = 92.16 μg/ml), C. sinuosa (IC50 = 112.44 μg/ml), and L. pinnatifida (IC50 = 115.11 μg/ml).

Keywords: α-glucosidase, lyophilized, macroalgae, spectrophotometrically

Procedia PDF Downloads 303

Authors: José Maria, Vânia Laranjo, Luís Abrunhosa, António Inês

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The occurrence of mycotoxigenic moulds such as Aspergillus, Penicillium and Fusarium in food and feed has an important impact on public health, by the appearance of acute and chronic mycotoxicoses in humans and animals, which is more severe in the developing countries due to lack of food security, poverty and malnutrition. This mould contamination also constitutes a major economic problem due the lost of crop production. A great variety of filamentous fungi is able to produce highly toxic secondary metabolites known as mycotoxins. Most of the mycotoxins are carcinogenic, mutagenic, neurotoxic and immunosuppressive, being ochratoxin A (OTA) one of the most important. OTA is toxic to animals and humans, mainly due to its nephrotoxic properties. Several approaches have been developed for decontamination of mycotoxins in foods, such as, prevention of contamination, biodegradation of mycotoxins-containing food and feed with microorganisms or enzymes and inhibition or absorption of mycotoxin content of consumed food into the digestive tract. Some group of Gram-positive bacteria named lactic acid bacteria (LAB) are able to release some molecules that can influence the mould growth, improving the shelf life of many fermented products and reducing health risks due to exposure to mycotoxins. Some LAB are capable of mycotoxin detoxification. Recently our group was the first to describe the ability of LAB strains to biodegrade OTA, more specifically, Pediococcus parvulus strains isolated from Douro wines. The pathway of this biodegradation was identified previously in other microorganisms. OTA can be degraded through the hydrolysis of the amide bond that links the L-β-phenylalanine molecule to the ochratoxin alpha (OTα) a non toxic compound. It is known that some peptidases from different origins can mediate the hydrolysis reaction like, carboxypeptidase A an enzyme from the bovine pancreas, a commercial lipase and several commercial proteases. So, we wanted to have a better understanding of this OTA degradation process when LAB are involved and identify which molecules where present in this process. For achieving our aim we used some bioinformatics tools (BLAST, CLUSTALX2, CLC Sequence Viewer 7, Finch TV). We also designed specific primers and realized gene specific PCR. The template DNA used came from LAB strains samples of our previous work, and other DNA LAB strains isolated from elderberry fruit, silage, milk and sausages. Through the employment of bioinformatics tools it was possible to identify several proteins belonging to the carboxypeptidase family that participate in the process of OTA degradation, such as serine type D-Ala-D-Ala carboxypeptidase and membrane carboxypeptidase. In conclusions, this work has identified carboxypeptidase proteins being one of the molecules present in the OTA degradation process when LAB are involved.

Keywords: carboxypeptidase, lactic acid bacteria, mycotoxins, ochratoxin a.

Procedia PDF Downloads 462

Authors: A. M. Tarola, R. Preti, A. M. Girelli, P. Campana

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Goat dairy products are gaining popularity worldwide. In developing countries, but also in many marginal regions of the Mediterranean area, goats represent a great part of the economy and ensure food security. In fact, these small ruminants are able to convert efficiently poor weedy plants and small trees into traditional products of high nutritional quality, showing great resilience to different climatic and environmental conditions. In developed countries, goat milk is appreciated for the presence of health-promoting compounds, bioactive compounds such as conjugated linoleic acids, oligosaccharides, sphingolipids and polyammines. This paper focuses on the recent advances in literature on the nutritional properties of goat milk and on innovative techniques to improve its quality as to become a promising functional food. The environmental sustainability of different methodologies of production has also been examined. Goat milk is valued today as a food of high nutritional value and functional properties as well as small environmental footprint. It is widely consumed in many countries due to high nutritional value, lower allergenic potential, and better digestibility when compared to bovine milk, that makes this product suitable for infants, elderly or sensitive patients. The main differences in chemical composition between a cow and goat milk rely on fat globules that in goat milk are smaller and in fatty acids that present a smaller chain length, while protein, fat, and lactose concentration are comparable. Milk nutritional properties have demonstrated to be strongly influenced by animal diet, genotype, and welfare, but also by season and production systems. Furthermore, there is a growing interest in the dairy industry in goat milk for its relatively high concentration of prebiotics and a good amount of probiotics, which have recently gained importance for their therapeutic potential. Therefore, goat milk is studied as a promising matrix to develop innovative functional foods. In addition to the economic and nutritional value, goat milk is considered a sustainable product for its small environmental footprint, as they require relatively little water and land, and less medical treatments, compared to cow, these characteristics make its production naturally vocated to organic farming. Organic goat milk production has becoming more and more interesting both for farmers and consumers as it can answer to several concerns like environment protection, animal welfare and economical sustainment of rural populations living in marginal lands. These evidences make goat milk an ancient food with novel properties and advantages to be valorized and exploited.

Keywords: goat milk, nutritional quality, bioactive compounds, sustainable production, animal welfare

Procedia PDF Downloads 149

Authors: Nabeeha Hassan Abdel Jalil, Robert Barigye, Hamda Al Alawi, Afra Al Dhaheri, Fatma Graiban Al Muhairi, Maryam Al Khateri, Nouf Al Alalawi, Susan Olet, Khaja Mohteshamuddin, Ahmad Al Aiyan, Mohamed Elfatih Hamad

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A serological survey was carried out to determine the seroprevalence of Brucella spp, Coxiella burnetii, Chlamydophila abortus, and Toxoplasma gondii in sheep and goat herds in the UAE. A total of 915 blood samples [n= 222, [sheep]; n= 215, [goats]) were collected from livestock farms in the Emirates of Abu Dhabi, Dubai, Sharjah and Ras Al-Khaimah (RAK). An additional 478 samples (n= 244, [sheep]; n= 234, (goats]) were collected from the Al Ain livestock central market and tested by indirect ELISA for pathogen-specific antibodies with the Brucella antibodies being further corroborated by the Rose-Bengal agglutination test. Seropositivity for the four pathogens is variably documented in sheep and goats from the study area. Respectively, the overall livestock farm prevalence rates for Brucella spp, C. burnetii, C. abortus, and T. gondii were 2.7%, 27.9%, 8.1%, and 16.7% for sheep, and 0.0%, 31.6%, 9.3%, and 5.1% for goats. Additionally, the seroprevalence rates Brucella spp, C. burnetii, C. abortus, and T. gondii in samples from the livestock market were 7.4%, 21.7%, 16.4%, and 7.0% for sheep, and 0.9%, 32.5%, 19.2%, and 11.1% for goats respectively. Overall, sheep had 12.59 more chances than goats of testing seropositive for Brucella spp (OR, 12.59 [95% CI 2.96-53.6]) but less likely to be positive for C. burnetii-antibodies (OR, 0.73 [95% CI 0.54-0.97]). Notably, the differences in the seroprevalence rates of C. abortus and T. gondii in sheep and goats were not statistically significant (p > 0.0500). The present data indicate that all the four study pathogens are present in sheep and goat populations in the UAE where coxiellosis is apparently the most seroprevalent followed by chlamydophilosis, toxoplasmosis, and brucellosis. While sheep from the livestock market were more likely than those from farms to be Brucella-seropositive than those, the overall exposure risk of C. burnetii appears to be greater for goats than sheep. As more animals from the livestock market were more likely to be seropositive to Chlamydophila spp, it is possible that under the UAE animal production conditions, at least, coxiellosis and chlamydophilosis are more likely to increase the culling rate of domesticated small ruminants than toxoplasmosis and brucellosis. While anecdotal reports have previously insinuated that brucellosis may be a significant animal health risk in the UAE, the present data suggest C. burnetii, C. abortus and T. gondii to be more significant pathogens of sheep and goats in the country. Despite this possibility, the extent to which these pathogens may nationally be contributing to reproductive failure in sheep and goat herds is not known and needs to be investigated. Potentially, these agents may also carry a potentially zoonotic risk that needs to be investigated in risk groups like farm workers, and slaughter house personnel. An ongoing study is evaluating the seroprevalence of bovine coxiellosis in the Emirate of Abu Dhabi and the data thereof will further elucidate on the broader epidemiological dynamics of the disease in the national herd.

Keywords: Brucella spp, Chlamydophila abortus, goat, sheep, Toxoplasma gondii, UAE

Procedia PDF Downloads 201

Authors: Nihan Nugay, Nur Cicek Kekec, Kalman Toth, Turgut Nugay, Joseph P. Kennedy

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In recent years, polyurethane structures are paving the way for elastomer usage in biology, human medicine, and biomedical application areas. Polyurethanes having a combination of high oxidative and hydrolytic stability and excellent mechanical properties are focused due to enhancing the usage of PUs especially for implantable medical device application such as cardiac-assist. Currently, unique polyurethanes consisting of polyisobutylenes as soft segments and conventional hard segments, named as PIB-based PUs, are developed with precise NCO/OH stoichiometry (∽1.05) for obtaining PIB-based PUs with enhanced properties (i.e., tensile stress increased from ∽11 to ∽26 MPa and elongation from ∽350 to ∽500%). Static and dynamic mechanical properties were optimized by examining stress-strain graphs, self-organization and crystallinity (XRD) traces, rheological (DMA, creep) profiles and thermal (TGA, DSC) responses. Annealing procedure was applied for PIB-based PUs. Annealed PIB-based PU shows ∽26 MPa tensile strength, ∽500% elongation, and ∽77 Microshore hardness with excellent hydrolytic and oxidative stability. The surface characters of them were examined with AFM and contact angle measurements. Annealed PIB-based PU exhibits the higher segregation of individual segments and surface hydrophobicity thus annealing significantly enhances hydrolytic and oxidative stability by shielding carbamate bonds by inert PIB chains. According to improved surface and microstructure characters, greater efforts are focused on analyzing protein adsorption and calcification profiles. In biomedical applications especially for cardiological implantations, protein adsorption inclination on polymeric heart valves is undesirable hence protein adsorption from blood serum is followed by platelet adhesion and subsequent thrombus formation. The protein adsorption character of PIB-based PU examines by applying Bradford assay in fibrinogen and bovine serum albumin solutions. Like protein adsorption, calcium deposition on heart valves is very harmful because vascular calcification has been proposed activation of osteogenic mechanism in the vascular wall, loss of inhibitory factors, enhance bone turnover and irregularities in mineral metabolism. The calcium deposition on films are characterized by incubating samples in simulated body fluid solution and examining SEM images and XPS profiles. PIB-based PUs are significantly more resistant to hydrolytic-oxidative degradation, protein adsorption and calcium deposition than ElastEonTM E2A, a commercially available PDMS-based PU, widely used for biomedical applications.

Keywords: biomedical application, calcification, polyisobutylene, polyurethane, protein adsorption

Procedia PDF Downloads 257

Authors: Marcos Silva Tavares, Jamile Raquel Regazzo, Edson José de Souza Sardinha, Murilo Mesquita Baesso

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Common beans are a widely cultivated and consumed legume globally, serving as a staple food for humans, especially in developing countries, due to their nutritional characteristics. Nitrogen (N) is the most limiting nutrient for productivity, and foliar analysis is crucial to ensure balanced nitrogen fertilization. Excessive N applications can cause, either isolated or cumulatively, soil and water contamination, plant toxicity, and increase their susceptibility to diseases and pests. However, the quantification of N using conventional methods is time-consuming and costly, demanding new technologies to optimize the adequate supply of N to plants. Thus, it becomes necessary to establish constant monitoring of the foliar content of this macronutrient in plants, mainly at the V4 stage, aiming at precision management of nitrogen fertilization. In this work, the objective was to evaluate the performance of a deep learning model, Resnet-50, in the classification of foliar nitrogen in common beans using RGB images. The BRS Estilo cultivar was sown in a greenhouse in a completely randomized design with four nitrogen doses (T1 = 0 kg N ha-1, T2 = 25 kg N ha-1, T3 = 75 kg N ha-1, and T4 = 100 kg N ha-1) and 12 replications. Pots with 5L capacity were used with a substrate composed of 43% soil (Neossolo Quartzarênico), 28.5% crushed sugarcane bagasse, and 28.5% cured bovine manure. The water supply of the plants was done with 5mm of water per day. The application of urea (45% N) and the acquisition of images occurred 14 and 32 days after sowing, respectively. A code developed in Matlab© R2022b was used to cut the original images into smaller blocks, originating an image bank composed of 4 folders representing the four classes and labeled as T1, T2, T3, and T4, each containing 500 images of 224x224 pixels obtained from plants cultivated under different N doses. The Matlab© R2022b software was used for the implementation and performance analysis of the model. The evaluation of the efficiency was done by a set of metrics, including accuracy (AC), F1-score (F1), specificity (SP), area under the curve (AUC), and precision (P). The ResNet-50 showed high performance in the classification of foliar N levels in common beans, with AC values of 85.6%. The F1 for classes T1, T2, T3, and T4 was 76, 72, 74, and 77%, respectively. This study revealed that the use of RGB images combined with deep learning can be a promising alternative to slow laboratory analyses, capable of optimizing the estimation of foliar N. This can allow rapid intervention by the producer to achieve higher productivity and less fertilizer waste. Future approaches are encouraged to develop mobile devices capable of handling images using deep learning for the classification of the nutritional status of plants in situ.

Keywords: convolutional neural network, residual network 50, nutritional status, artificial intelligence

Procedia PDF Downloads 19

Authors: Amal Balila, Maria Vahdati

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Earth is a green building material with very low embodied energy and almost zero greenhouse gas emissions. However, it lacks strength and durability in its natural state. By responsibly sourcing stabilisers, it's possible to enhance its strength. This research draws inspiration from the robustness of termite mounds, where termites incorporate glycoproteins from their saliva during construction. Biomimicry explores the potential of these termite stabilisers in producing bio-inspired adobe bricks. The meat industry generates significant waste during slaughter, including blood, skin, bones, tendons, gastrointestinal contents, and internal organs. While abundant, many meat by-products raise concerns regarding human consumption, religious orders, cultural and ethical beliefs, and also heavily contribute to environmental pollution. Extracting and utilising proteins from this waste is vital for reducing pollution and increasing profitability. Exploring the untapped potential of meat industry waste, this research investigates how glycoproteins could revolutionize adobe brick construction. Bovine serum albumin (BSA) from cows' blood and mucin from porcine stomachs were the chosen glycoproteins used as stabilisers for adobe brick production. Despite their wide usage across various fields, they have very limited utilisation in food processing. Thus, both were identified as potential stabilisers for adobe brick production in this study. Two soil types were utilised to prepare adobe bricks for testing, comparing controlled unstabilised bricks with glycoprotein-stabilised ones. All bricks underwent testing for unconfined compressive strength and erosion resistance. The primary finding of this study is the efficacy of BSA, a glycoprotein derived from cows' blood and a by-product of the beef industry, as an earth construction stabiliser. Adding 0.5% by weight of BSA resulted in a 17% and 41% increase in the unconfined compressive strength for British and Sudanese adobe bricks, respectively. Further, adding 5% by weight of BSA led to a 202% and 97% increase in the unconfined compressive strength for British and Sudanese adobe bricks, respectively. Moreover, using 0.1%, 0.2%, and 0.5% by weight of BSA resulted in erosion rate reductions of 30%, 48%, and 70% for British adobe bricks, respectively, with a 97% reduction observed for Sudanese adobe bricks at 0.5% by weight of BSA. However, mucin from the porcine stomach did not significantly improve the unconfined compressive strength of adobe bricks. Nevertheless, employing 0.1% and 0.2% by weight of mucin resulted in erosion rate reductions of 28% and 55% for British adobe bricks, respectively. These findings underscore BSA's efficiency as an earth construction stabiliser for wall construction and mucin's efficacy for wall render, showcasing their potential for sustainable and durable building practices.

Keywords: biomimicry, earth construction, industrial waste management, sustainable building materials, termite mounds.

Procedia PDF Downloads 51

Authors: Bipasa Dey, Varsha Panwar, Tanmay Dutta

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Laccase, a multicopper oxidase (EC 1.10.3.2) have been at the forefront as a superior industrial biocatalyst. They are versatile in terms of bestowing sustainable and ecological catalytic reactions such as polymerisation, xenobiotic degradation and bioremediation of phenolic and non-phenolic compounds. Regardless of the wide biotechnological applications, the critical limiting factors viz. reusability, retrieval, and storage stability still prevail. This can cause an impediment in their applicability. Crosslinked enzyme aggregates (CLEAs) have emerged as a promising technique that rehabilitates these essential facets, albeit at the expense of their enzymatic activity. The carrier free crosslinking method prevails over the carrier-bound immobilisation in conferring high productivity, low production cost owing to the absence of additional carrier and circumvent any non-catalytic ballast which could dilute the volumetric activity. To the best of our knowledge, the ε-amino group of lysyl residue is speculated as the best choice for forming Schiff’s base with glutaraldehyde. Despite being most preferrable, excess glutaraldehyde can bring about disproportionate and undesirable crosslinking within the catalytic site and hence could deliver undesirable catalytic losses. Moreover, the surface distribution of lysine residues in Trametes versicolor laccase is significantly less. Thus, to mitigate the adverse effect of glutaraldehyde in conjunction with scaling down the degradation or catalytic loss of the enzyme, crosslinking with inert substances like gelatine, collagen, Bovine serum albumin (BSA) or excess lysine is practiced. Analogous to these molecules, sugars have been well known as a protein stabiliser. It helps to retain the structural integrity, specifically secondary structure of the protein during aggregation by changing the solvent properties. They are comprehended to avert protein denaturation or enzyme deactivation during precipitation. We prepared crosslinked enzyme aggregates (CLEAs) of laccase from T. versicolor with the aid of sugars. The sugar CLEAs were compared with the classic BSA and glutaraldehyde laccase CLEAs concerning physico-chemical properties. The activity recovery for the fructose CLEAs were found to be ~20% higher than the non-sugar CLEA. Moreover, the 𝐾𝑐𝑎𝑡𝐾𝑚⁄ values of the CLEAs were two and three-fold higher than BSA-CLEA and GACLEA, respectively. The half-life (t1/2) deciphered by sugar-CLEA was higher than the t1/2 of GA-CLEAs and free enzyme, portraying more thermal stability. Besides, it demonstrated extraordinarily high pH stability, which was analogous to BSA-CLEA. The promising attributes of increased storage stability and recyclability (>80%) gives more edge to the sugar-CLEAs over conventional CLEAs of their corresponding free enzyme. Thus, sugar-CLEA prevails in furnishing the rudimentary properties required for a biocatalyst and holds many prospects.

Keywords: cross-linked enzyme aggregates, laccase immobilization, enzyme reusability, enzyme stability

Procedia PDF Downloads 102

Authors: Abdirahman Barre, Abdihamid Salad Hassan, Iftin Abdi Mohamud, Abdirahman Mohamed Mohamud, Ahmed Adan Mohamed, Mukhtaar Mohamed Idow

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Pasteurellosis (Hemorrhagic septicemia) is a common respiratory disease of camel that is an acutely fatal disease caused by Pasteurella multocida type A or several serotypes of Mannheimia hemolytic, which also affect other animals. The disease had shown to spread between animals, across herds and to humans. Meaning that the disease is Zoonosis. The study aimed at establishment of sero-prevalence of Pasteurellosis in some selected Districts of camel rearing in the Benadir Region. It was a cross-sectional study, where the study population was purposively chosen to consist of animals taken within three sub-Districts of Benadir Region, namely Sub-District (Daynile Township), Sub-District (Yaaqshid) Sub-District (kaxda). This was because they normally handle many camels in a day, thus making it easy for the investigator to access the required number conveniently; it was also assumed that data collected from these for-slaughter camels was representative of the situation in the sub-District/county. A total of one hundred and sixty camels were tested using four serological tests: Rose Bengal Plate Test (RBPT),) and Complex Fixation Test (CFT). The serological tests were purposively chosen to increase the chances of picking positive cases and also to compare their sensitivities with respect to camel serum since they were originally meant for use on bovine serum. Blood samples (15 ml) were collected for serum harvesting from the jugular veins of the animals as they were waiting to be examined. Rose Bengal plate test and CFT were run at a laboratory within the Department of Veterinary Medicine, University of Horsed, 21 October campus; serum samples having been transported in a cool box. On average, out of an overall total of 300 serum samples tested, 180 samples were selected as sample procedures and were given eleven (11) positive results, amounting to a prevalence of 6.67%. For the three Districts, respective prevalence (averaged from the two (2) serological tests run) were: 7% (3/50) for Yaqshiid; 8% (3/60) for Deyniile and 10% (3/70) for Kaxda. When sensitivities of the two (2) serological tests were compared, there was no significant difference between them with respect to the picking of positive cases (p=0.05). The study has demonstrated presence of Pasterolosis in camels in Benadir Region and the authors are recommending the usage of RBPT and CFT as screening tests, since they are cheap, quick, and easy to carry out. Any of the other three involving tests can then be used if one wants to establish respective titers. Therefore, further detailed investigation needs to be conducted so as to understand specific etiological agents causing pasteurollosis in camel and can be instituted to optimize the benefit obtained from the camel sector.

Keywords: hemorrhagic septicemia, camel, prevalence, Benadir region, Somalia

Procedia PDF Downloads 72

Authors: Eva Filová, Jana Daňková, Věra Sovková, Matej Daniel

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Titanium alloys are biocompatible metals that are widely used in clinical practice as load bearing implants. The chemical modification may influence cell adhesion, proliferation, and differentiation as well as stiffness of the material. The aim of the study was to evaluate the adhesion, growth and differentiation of pig mesenchymal stem cells on the novel beta titanium alloy Ti36Nb6Ta compared to standard medical titanium alloy Ti6Al4V. Discs of Ti36Nb6Ta and Ti6Al4V alloy were sterilized by ethanol, put in 48-well plates, and seeded by pig mesenchymal stem cells at the density of 60×103/cm2 and cultured in Minimum essential medium (Sigma) supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cell viability was evaluated using MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay;Promega), cell proliferation using Quant-iT™ ds DNA Assay Kit (Life Technologies). Cells were stained immunohistochemically using monoclonal antibody beta-actin, and secondary antibody conjugated with AlexaFluor®488 and subsequently the spread area of cells was measured. Cell differentiation was evaluated by alkaline phosphatase assay using p-nitrophenyl phosphate (pNPP) as a substrate; the reaction was stopped by NaOH, and the absorbance was measured at 405 nm. Osteocalcin, specific bone marker was stained immunohistochemically and subsequently visualized using confocal microscopy; the fluorescence intensity was analyzed and quantified. Moreover, gene expression of osteogenic markers osteocalcin and type I collagen was evaluated by real-time reverse transcription-PCR (qRT-PCR). For statistical evaluation, One-way ANOVA followed by Student-Newman-Keuls Method was used. For qRT-PCR, the nonparametric Kruskal-Wallis Test and Dunn's Multiple Comparison Test were used. The absorbance in MTS assay was significantly higher on titanium alloy Ti6Al4V compared to beta titanium alloy Ti36Nb6Ta on days 7 and 14. Mesenchymal stem cells were well spread on both alloys, but no difference in spread area was found. No differences in alkaline phosphatase assay, fluorescence intensity of osteocalcin as well as the expression of type I collagen, and osteocalcin genes were observed. Higher expression of type I collagen compared to osteocalcin was observed for cells on both alloys. Both beta titanium alloy Ti36Nb6Ta and titanium alloy Ti6Al4V Ti36Nb6Ta supported mesenchymal stem cellsˈ adhesion, proliferation and osteogenic differentiation. Novel beta titanium alloys Ti36Nb6Ta is a promising material for bone implantation. The project was supported by the Czech Science Foundation: grant No. 16-14758S, the Grant Agency of the Charles University, grant No. 1246314 and by the Ministry of Education, Youth and Sports NPU I: LO1309.

Keywords: beta titanium, cell growth, mesenchymal stem cells, titanium alloy, implant

Procedia PDF Downloads 316

Authors: Busra Gundede, Ozal Mutlu, Nagihan Gulsoy

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Background: Over the years, material research has led to the development of numerous metals and alloys for using in biomedical applications. One of the major tasks of biomaterial research is the functionalization of the material surface to improve the biocompatibility according to a specific application. 316L and 316L alloys are excellent for various bio-applications. This research was investigated the effect of titanium (Ti), niobium (Nb), and zirconium (Zr) additives on injection molded austenitic grade 316L stainless steels in vitro biocompatibility. For this purpose, cytotoxic tests were performed to evaluate the potential biocompatibility of the specimens. Materials and Methods: 3T3 fibroblast were cultivated in DMEM supplemented with 10% fetal bovine serum and %1 penicillin-streptomycin at 37°C with 5% CO2 and 95%humidity. Trypsin/EDTA solution was used to remove cells from the culture flask. Cells were reseeded at a density of 1×105cell in 25T flasks. The medium change took place every 3 days. The trypan blue assay was used to determine cell viability. Cell viability is calculated as the number of viable cells divided by the total number of cells within the grids on the cell counter machine counted the number of blue staining cells and the number of total cells. Cell viability should be at least 95% for healthy log-phase cultures. MTT assay was assessed for 96-hours. Cells were cultivated in 6-well flask within 5 ml DMEM and incubated as same conditions. 0,5mg/ml MTT was added for 4-hours and then acid-isoprohanol was added for solubilize to formazan crystals. Cell morphology after 96h was investigated by SEM. The medium was removed, samples were washed with 0.15 M PBS buffer and fixed for 12h at 4- 8°C with %2,5 gluteraldehyte. Samples were treated with 1% osmium tetroxide. Samples were then dehydrated and dried, mounted on appropriate stubs with colloidal silver and sputter-coated with gold. Images were collected using a scanning electron microscope. ROS assay is a cell viability test for in vitro studies. Cells were grown for 96h, ROS solution added on cells in 6 well plate flask and incubated for 1h. Fluorescence signal indicates ROS generation by cells. Results: Trypan Blue exclusion assay results were 96%, 92%, 95%, 90%, 91% for negative control group, 316L, 316L-Ti, 316L-Nb and 316L-Zr, respectively. Results were found nearly similar to each other when compared with control group. Cell viability from MTT analysis was found to be 100%, 108%, 103%, 107%, and 105% for the control group, 316L, 316L-Ti, 316L-Nb and 316L-Zr, respectively. Fluorescence microscopy analysis indicated that all test groups were same as the control group in ROS assay. SEM images demonstrated that the attachment of 3T3 cells on biomaterials. Conclusion: We, therefore, concluded that Ti, Nb and Zr additives improved physical properties of 316L stainless. In our in vitro experiments showed that these new additives did not modify the cytocompatibility of stainless steel and these additives on 316L might be useful for biomedical applications.

Keywords: 316L stainles steel, biocompatibility, cell culture, Ti, Nb, Zr

Procedia PDF Downloads 513

Authors: Raj Kishore Swain, Kamdev Sethy, Sumanta Kumar Mishra

Abstract:

Selenium is an essential trace element for the chicken with a variety of biological functions like growth, fertility, immune system, hormone metabolism, and antioxidant defense systems. Selenium deficiency in chicken causes exudative diathesis, pancreatic dystrophy and nutritional muscle dystrophy of the gizzard, heart and skeletal muscle. Additionally, insufficient immunity, lowering of production ability, decreased feathering of chickens and increased embryo mortality may occur due to selenium deficiency. Nano elemental selenium, which is bright red, highly stable, soluble and of nano meter size in the redox state of zero, has high bioavailability and low toxicity due to the greater surface area, high surface activity, high catalytic efficiency and strong adsorbing ability. To assess the effect of dietary nano-Se on performance and expression of gene in Vencobb broiler birds in comparison to its inorganic form (sodium selenite), four hundred fifty day-old Vencobb broiler chicks were randomly distributed into 9 dietary treatment groups with two replicates with 25 chicks per replicate. The dietary treatments were: T1 (Control group): Basal diet; T2: Basal diet with 0.3 ppm of inorganic Se; T3: Basal diet with 0.01875 ppm of nano-Se; T4: Basal diet with 0.0375 ppm of nano-Se; T5: Basal diet with 0.075 ppm of nano-Se, T6: Basal diet with 0.15 ppm of nano-Se, T7: Basal diet with 0.3 ppm of nano-Se, T8: Basal diet with 0.60 ppm of nano-Se, T9: Basal diet with 1.20 ppm of nano-Se. Nano selenium was synthesized by mixing sodium selenite with reduced glutathione and bovine serum albumin. The experiment was carried out in two phases: starter phase (0-3 wks), finisher phase (4-5 wk) in deep litter system. The body weight at the 5th week was best observed in T4. The best feed conversion ratio at the end of 5th week was observed in T4. Erythrocytic catalase, glutathione peroxidase and superoxide dismutase activity were significantly (P < 0.05) higher in all the nano selenium treated groups at 5th week. The antibody titers (log2) against Ranikhet diseases vaccine immunization of 5th-week broiler birds were significantly higher (P < 0.05) in the treatments T4 to T7. The selenium levels in liver, breast, kidney, brain, and gizzard were significantly (P < 0.05) increased with increasing dietary nano-Se indicating higher bioavailability of nano-Se compared to inorganic Se. The real time polymer chain reaction analysis showed an increase in the expression of antioxidative gene in T4 and T7 group. Therefore, it is concluded that supplementation of nano-selenium at 0.0375 ppm over and above the basal level can improve the body weight, antioxidant enzyme activity, Se bioavailability and expression of the antioxidative gene in broiler birds.

Keywords: chicken, growth, immunity, nano selenium

Procedia PDF Downloads 177

Authors: Rossana Roila, Raffaella Branciari, Lorenzo Cardinali, David Ranucci

Abstract:

The wild boar (Sus scrofa) population has strongly increased across Europe in the last decades, also causing severe fauna management issues. In central Italy, wild boar is the main hunted wild game species, with approximately 40,000 animals killed per year only in the Umbria region. The meat of the game is characterized by high-quality nutritional value as well as peculiar taste and aroma, largely appreciated by consumers. This type of meat and products thereof can meet the current consumers’ demand for higher quality foodstuff, not only from a nutritional and sensory point of view but also in relation to environmental sustainability, the non-use of chemicals, and animal welfare. The game meat production chain is characterized by some gaps from a hygienic point of view: the harvest process is usually conducted in a wild environment where animals can be more easily contaminated during hunting and subsequent practices. The definition and implementation of a certified and controlled supply chain could ensure quality, traceability and safety for the final consumer and therefore promote game meat products. According to European legislation in some animal species, such as bovine, the use of weak acid solutions for carcass decontamination is envisaged in order to ensure the maintenance of optimal hygienic characteristics. A preliminary study was carried out to evaluate the applicability of similar strategies to control the hygienic level of wild boar carcasses. The carcasses, harvested according to the selective method and processed into the game-handling establishment, were treated by nebulization with two different solutions: a 2% food-grade lactic acid solution and aromatic vinegar. Swab samples were performed before treatment and in different moments after-treatment of the carcasses surfaces and subsequently tested for Total Aerobic Mesophilic Load, Total Aerobic Psychrophilic Load, Enterobacteriaceae, Staphylococcus spp. and lactic acid bacteria. The results obtained for the targeted microbial populations showed a positive effect of the application of the lactic acid solution on all the populations investigated, while aromatic vinegar showed a lower effect on bacterial growth. This study could lay the foundations for the optimization of the use of a lactic acid solution to treat wild boar carcasses aiming to guarantee good hygienic level and safety of meat.

Keywords: game meat, food safety, process hygiene criteria, microbial population, microbial growth, food control

Procedia PDF Downloads 158

Authors: M. Shahin Alam, Satoru Takahashi, Mariko Itoh, Miyuki Komura, Mayuko Suzuki, Natthanan Sangsriratanakul, Kazuaki Takehara

Abstract:

Cleaning and disinfection are key components of routine biosecurity in livestock farming and food processing industry. The usage of suitable disinfectants and their proper concentration are important factors for a successful biosecurity program. Disinfectants have optimum bactericidal and virucidal efficacies at temperatures above 20°C, but very few studies on application and effectiveness of disinfectants at low temperatures have been done. In the present study, the bactericidal efficacies of food additive grade calcium hydroxide (FdCa(OH)), quaternary ammonium compound (QAC) and their mixture, were investigated under different conditions, including time, organic materials (fetal bovine serum: FBS) and temperature, either in suspension or in carrier test. Salmonella Infantis and Escherichia coli, which are the most prevalent gram negative bacteria in commercial poultry housing and food processing industry, were used in this study. Initially, we evaluated these disinfectants at two different temperatures (4°C and room temperature (RT) (25°C ± 2°C)) and 7 contact times (0, 5 and 30 sec, 1, 3, 20 and 30 min), with suspension tests either in the presence or absence of 5% FBS. Secondly, we investigated the bactericidal efficacies of these disinfectants by carrier tests (rubber, stainless steel and plastic) at same temperatures and 4 contact times (30 sec, 1, 3, and 5 min). Then, we compared the bactericidal efficacies of each disinfectant within their mixtures, as follows. When QAC was diluted with redistilled water (dW2) at 1: 500 (QACx500) to obtain the final concentration of didecyl-dimethylammonium chloride (DDAC) of 200 ppm, it could inactivate Salmonella Infantis within 5 sec at RT either with or without 5% FBS in suspension test; however, at 4°C it required 30 min in presence of 5% FBS. FdCa(OH)2 solution alone could inactivate bacteria within 1 min both at RT and 4°C even with 5% FBS. While FdCa(OH)2 powder was added at final concentration 0.2% to QACx500 (Mix500), the mixture could inactivate bacteria within 30 sec and 5 sec, respectively, with or without 5% FBS at 4°C. The findings from the suspension test indicated that low temperature inhibited the bactericidal efficacy of QAC, whereas Mix500 was effective, regardless of short contact time and low temperature, even with 5% FBS. In the carrier test, single disinfectant required bit more time to inactivate bacteria on rubber and plastic surfaces than on stainless steel. However, Mix500 could inactivate S. Infantis on rubber, stainless steel and plastic surfaces within 30 sec and 1 min, respectively, at RT and 4°C; but, for E. coli, it required only 30 sec at both temperatures. So, synergistic effects were observed on different carriers at both temperatures. For a successful enhancement of biosecurity during winter, the disinfectants should be selected that could have short contact times with optimum efficacy against the target pathogen. The present study findings help farmers to make proper strategies for application of disinfectants in their livestock farming and food processing industry.

Keywords: carrier, food additive grade calcium hydroxide (FdCa(OH)₂), quaternary ammonium compound, synergistic effects

Procedia PDF Downloads 294

Authors: Daniel Dahis, Haim Azhari

Abstract:

Noninvasive image-guided intracranial treatments using high intensity focused ultrasound (HIFU) are on the course of translation into clinical applications. They include, among others, tumor ablation, hyperthermia, and blood-brain-barrier (BBB) penetration. Since many of these procedures are associated with local temperature elevation, thermal monitoring is essential. MRI constitutes an imaging method with high spatial resolution and thermal mapping capacity. It is the currently leading modality for temperature guidance, commonly under the name MRgHIFU (magnetic-resonance guided HIFU). Nevertheless, MRI is a very expensive non-portable modality which jeopardizes its accessibility. Ultrasonic thermal monitoring, on the other hand, could provide a modular, cost-effective alternative with higher temporal resolution and accessibility. In order to assess the feasibility of ultrasonic brain thermal monitoring, this study investigated the usage of brain tissue attenuation coefficient (AC) temporal changes as potential estimators of thermal changes. Newton's law of cooling describes a temporal exponential decay behavior for the temperature of a heated object immersed in a relatively cold surrounding. Similarly, in the case of cerebral HIFU treatments, the temperature in the region of interest, i.e., focal zone, is suggested to follow the same law. Thus, it was hypothesized that the AC of the irradiated tissue may follow a temporal exponential behavior during cool down regime. Three ex-vivo bovine brain tissue specimens were inserted into plastic containers along with four thermocouple probes in each sample. The containers were placed inside a specially built ultrasonic tomograph and scanned at room temperature. The corresponding pixel-averaged AC was acquired for each specimen and used as a reference. Subsequently, the containers were placed in a beaker containing hot water and gradually heated to about 45ᵒC. They were then repeatedly rescanned during cool down using ultrasonic through-transmission raster trajectory until reaching about 30ᵒC. From the obtained images, the normalized AC and its temporal derivative as a function of temperature and time were registered. The results have demonstrated high correlation (R² > 0.92) between both the brain AC and its temporal derivative to temperature. This indicates the validity of the hypothesis and the possibility of obtaining brain tissue temperature estimation from the temporal AC thermal changes. It is important to note that each brain yielded different AC values and slopes. This implies that a calibration step is required for each specimen. Thus, for a practical acoustic monitoring of the brain, two steps are suggested. The first step consists of simply measuring the AC at normal body temperature. The second step entails measuring the AC after small temperature elevation. In face of the urging need for a more accessible thermal monitoring technique for brain treatments, the proposed methodology enables a cost-effective high temporal resolution acoustical temperature estimation during HIFU treatments.

Keywords: attenuation coefficient, brain, HIFU, image-guidance, temperature

Procedia PDF Downloads 161