Search results for: bio-activity

Authors: Abdul Matin, Muhammad Ajmal, Uzma Yunus, Noaman-ul Haq, Hafiz M. Shohaib, Ambreen G. Muazzam

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Carbon nanoparticles (CNPs) have unique properties that are useful for the diagnosis and treatment of cancer due to their precise properties like small size (ideal for delivery within the body) stability in solvent and tunable surface chemistry for targeted delivery. Here, highly fluorescent, monodispersed and water-soluble CNPs were synthesized directly from a suitable carbohydrate source (glucose and sucrose) by one-step acid assisted ultrasonic treatment at 35 KHz for 4 hours. This method is green, simple, rapid and economical and can be used for large scale production and applications. The average particle sizes of CNPs are less than 10nm and they emit bright and colorful green-blue fluorescence under the irradiation of UV-light at 365nm. The CNPs were characterized by scanning electron microscopy, fluorescent spectrophotometry, Fourier transform infrared spectrophotometry, ultraviolet-visible spectrophotometry and TGA analysis. Fluorescent CNPs were used as fluorescent probe and nano-carriers for anticancer drug. Functionalized CNPs (with ethylene diamine) were attached with anticancer drug-Methotrexate. In vitro bioactivity and biocompatibility of CNPs-drug conjugates was evaluated by LDH assay and Sulforhodamine B assay using human lung carcinoma cell lines (H157). Our results reveled that CNPs showed biocompatibility and CNPs-anticancer drug conjugates have shown potent cytotoxic effects and high antitumor activities in lung cancer cell lines. CNPs are proved to be excellent substitute for conventional drug delivery cargo systems and anticancer therapeutics in vitro. Our future studies will be more focused on using the same nanoparticles in vivo model system.

Keywords: carbon nanoparticles, carbon nanoparticles-methotrexate conjugates, human lung carcinoma cell lines, lactate dehydrogenase, methotrexate

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Authors: S. Mamoucha, N. Tsafantakis, Α. Ioannidis, S. Chatzipanagiotou, C. Nikolaou, L. Skaltsounis, N. Fokialakis, N. Christodoulakis

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Introduction: Plant secondary metabolites (SM) can be grouped into three chemically distinct groups: terpenes, phenolics, and nitrogen-containing compounds. For many years the adaptive significance of SM was unknown. They were thought to be functionless end-products. Currently it is accepted that many secondary metabolites (also known as natural products) have important ecological roles in plants. For instance, they serve as attractants (odor, color, taste) for pollinators and seed-dispersing animals. Moreover, they protect plants from herbivores, microbial pathogens and from environmental stress (high and low temperatures, drought, alkalinity, salinity, radiation etc). It is well known that both biotic and abiotic stress often increase the accumulation of SM. The local climatic conditions, seasonal changes, external factors such as light, temperature, humidity affect the biosynthesis and composition of secondary metabolites. A well known dioecious evergreen plant, Pistacia lentiscus L. (mastic tree), was selected in order to study the metabolic variations occur in response to the different climate conditions, due to the seasonal variation and its effect on the biosynthesis of bioactive compounds. Materials-methods: Young and mature leaves were collected in January and July 2014, dried and extracted by accelerated solvent extraction (Dionex ASE™ 350) using solvents of increased polarity (DCM, MeOH, and H2O). GC-MS and UHPLC-HRMS analysis were carried out in order to define the nature and the relative abundance of SM. The antibacterial activity was evaluated by using the Agar Disc Diffusion Assay against ATCC and clinical isolates strains: Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Streptococcus mutans and Klebsiella pneumoniae. All tests were carried out in duplicate and the average radii of the inhibition zones were calculated for each extract. Results: According to the phytochemical profile obtained from each extract, the biosynthesis of SM varied both qualitatively and quantitatively under the two different types of seasonal stress. With exception of the biologically inactive nonpolar DCM extract of July, all extracts inhibited the growth of most of the investigated microorganisms. A clear positive correlation has been observed between the relative abundance of SM and the bioactivity of the DCM extracts of January and July. Observed changes during phytochemical analysis were mainly focused on the triterpenoid content. On the other hand, the bioactivity of the polar extracts (MeOH and H2O) of January and July resulted practically invariable against most of the microorganisms, besides the significant variation of the SM content due to the seasonal variation. Conclusion: Our results clearly confirmed the hypothesis of abiotic stress as an important regulating factor that significantly affects the biosynthesis of secondary metabolites and thus the presence of bioactive compounds. Acknowledgment: This work was supported by IKY - State Scholarship Foundation, Athens, Greece.

Keywords: antibacterial screening, phytochemical profile, Pistacia lentiscus, abiotic stress

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Authors: Geminne Manzano, Porfirio Aliño, Clairecynth Yu, Lilibeth Salvador-Reyes, Viviene Santiago

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Many marine benthic organisms produce secondary metabolites that serve as ecological roles to different biological and environmental factors. The secondary metabolites found in these organisms like algae, sponges, tunicates and worms exhibit variation at different scales. Understanding the chemical variation can be essential in deriving the evolutionary and ecological function of the secondary metabolites that may explain their patterns. Ecological surveys were performed on two collection sites representing from two Philippine marine biogeographic regions – in Oriental Mindoro located on the West Philippine Sea (WPS) and in Zamboanga del Sur located at Celebes Sea (CS), where a total of 39 Xestospongia sp. sponges were collected using SCUBA. The sponge samples were transported to the laboratory for taxonomic identification and chemical analysis. Biological and environmental factors were investigated to determine their relation to the abundance and distribution patterns and its spatial variability of their secondary metabolite production. Extracts were subjected to thin-layer chromatography and anti-proliferative assays to confirm the presence of Renieramycin-M and to test its cytotoxicity. The blue sponges were found to be more abundant on the WPS than in CS. Both the benthic community and the fish community in Oriental Mindoro, WPS and Zamboanga del Sur, CS sites are characterized by high species diversity and abundance and a very high biomass category. Environmental factors like depth and monsoonal exposure were also compared showing that wave exposure and depth are associated with the abundance and distribution of the sponges. Renieramycin-M presence using the TLC profiles between the sponge extracts from WPS and from CS showed differences in the Reniermycin-M presence and the presence of other functional groups were observed between the two sites. In terms of bioactivity, different responses were also exhibited by the sponge extracts coming from the different region. Different responses were also noted on its bioactivity depending on the cell lines tested. Exploring the influence of ecological parameters on the chemical variation can provide deeper chemical ecological insights in the knowledge and their potential varied applications at different scales. The results of this study provide further impetus in pursuing studies into patterns and processes of the chemical diversity of the Philippine blue sponge, Xestospongia sp. and the chemical ecological significance of the coral triangle.

Keywords: chemical ecology, porifera, renieramycin-m, spatial variability, Xestospongia sp.

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Authors: Z. Terzopoulou, I. Koliakou, D. Bikiaris

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Tissue engineering offers a new approach to regenerate diseased or damaged tissues such as bone. Great effort is devoted to eliminating the need of removing non-degradable implants at the end of their life span, with biodegradable polymers playing a major part. Poly(ε-caprolactone) (PCL) is one of the best candidates for this purpose due to its high permeability, good biodegradability and exceptional biocompatibility, which has stimulated extensive research into its potential application in the biomedical fields. However, PCL degrades much slower than other known biodegradable polymers and has a total degradation of 2-4 years depending on the initial molecular weight of the device. This is due to its relatively hydrophobic character and high crystallinity. Consequently, much attention has been given to the tunable degradation of PCL to meet the diverse requirements of biomedicine. Poly(ε-caprolactone) (PCL) is a biodegradable polyester that lacks bioactivity, so when used in bone tissue engineering, new bone tissue cannot bond tightly on the polymeric surface. Therefore, it is important to incorporate reinforcing fillers into PCL matrix in order to result in a promising combination of bioactivity, biodegradability, and strength. Natural clay halloysite nanotubes (HNTs) were incorporated into PCL polymeric matrix, via in situ ring-opening polymerization of caprolactone, in concentrations 0.5, 1 and 2.5 wt%. Both unmodified and modified with aminopropyltrimethoxysilane (APTES) HNTs were used in this study. The effect of nanofiller concentration and functionalization with end-amino groups on the physicochemical properties of the prepared nanocomposites was studied. Mechanical properties were found enhanced after the incorporation of nanofillers, while the modification increased further the values of tensile and impact strength. Thermal stability of PCL was not affected by the presence of nanofillers, while the crystallization rate that was studied by Differential Scanning Calorimetry (DSC) and Polarized Light Optical Microscopy (POM) increased. All materials were subjected to enzymatic hydrolysis in phosphate buffer in the presence of lipases. Due to the hydrophilic nature of HNTs, the biodegradation rate of nanocomposites was higher compared to neat PCL. In order to confirm the effect of hydrophilicity, contact angle measurements were also performed. In vitro biomineralization test confirmed that all samples were bioactive as mineral deposits were detected by X-ray diffractometry after incubation in SBF. All scaffolds were tested in relevant cell culture using osteoblast-like cells (MG-63) to demonstrate their biocompatibility

Keywords: biomaterials, nanocomposites, scaffolds, tissue engineering

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Authors: Zachary Muthii Rukenya, James Mbaria, Peter Mbaabu, Kiama Stephen Gitahi, Ronald Onzago

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Aloe turkanensis is one of the widely used medicinal shrub and in Kenya the plant is mainly found in Baringo, Isiolo, Laikipia, Turkana and West Pokot Counties where it is used in ethno-medicine and ethno-veterinary medicine. The Turkana community uses the plant products to manage malaria, wounds, stomach ache, constipation, pain, skin infection, poultry diseases and retained afterbirth in cows. This evaluated the efficacy and safety of the plant obtained from Turkana County, Kenya. Preliminary data on the use of the plant in the county was collected through observation, photographing and interviews. A sample of the whole plant was harvested in Natira sublocation, in ex-Turkana west district in February 2012 after identification by Aloe-working group herbalists who voluntarily provided information on its medicinal uses. Botanical identification was done at Kenya Forest Research Centre in Karura where voucher specimen was deposited. Cold maceration using 70% methanol and distilled water was used for extraction. Bioassays were to determine the effects of the plant extracts on brine shrimp and selected bacterial and fungal cultures. The extracts were tested in-vitro activity against standard cultures of B. cereus (ATCC 11778), S. aureus (ATCC25923), P. aeroginosa (ATCC 27853), E. coli (ATCC 25922) and a human infections clinical isolate of C. albicans. The extracts of Aloe turkanensis inhibited the growth B. cereus (100-200 mg/ml), S. aureus (50-100 mg/ml), P. aeroginosa (200mg/ml), E. coli (400mg/ml) while C. albicans was not affected. The extracts also inhibited the growth of S. aureus and B. cereus with mean diameters of inhibition zones being 19.75±1 mm and 18.5±05 mm reapectively. Phytochemical screening showed the presence of alkaloids, tarpenoids, steroids, quinones, saponins and tannins in the plant extracts. The extract was found to be non-toxic at a concentration of 1000µg/ml with a 100% survival of Brine Shrimp larva. It was concluded that Aloe turkanensis growing the study area has metabolites that inhibit the growth of microorganisms and is however, there is need for further studies to validate the in-vivo bioactivity of the plant and more generate adequate toxicological data.to support conservation, value chain addition of its products and widespread use as a herbal remedy.

Keywords: Aloe turkanensis, bioactivity, cultivated, human infections

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Authors: Roihatul Mutiah, Sukardiman, Aty Widyawaruyanti

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Calotropis gigantiea (L.) R. Br (Apocynaceae) commonly called as “Biduri” or “giant milk weed” is a well-known weed to many cultures for treating various disorders. Several studies reported that C.gigantea roots has anticancer activity. The main aim of this research was to isolate and identify an active marker compound of C.gigantea roots for quality control purpose of its extract in the development as anticancer natural product. The isolation methods was bioactivity guided column chromatography, TLC, and HPLC. Evaluated anticancer activity of there substances using MTT assay methods. Identification structure active compound by UV, 1HNMR, 13CNMR, HMBC, HMQC spectral and other references. The result showed that the marker active compound was identical as Calotropin.

Keywords: calotropin, Calotropis gigantea, anticancer, marker active

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Authors: Uftah Ali M. Shushni, Viola Stuppec, Ulrike Lindequist

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Because of the evolving resistance of microorganisms to existing antibiotics, there is an increasing need for new antibiotics not only in human but also in veterinary medicine. As part of our ongoing work on the secondary metabolites produced by marine fungi, the organic extract of the culture filtrate of an Ascomycetous fungus, which was found on driftwood collected from the coast of the Greifswalder Bodden, Baltic Sea, Germany displayed antimicrobial activity against some fish and human pathogenic bacteria. Bioactivity-guided column chromatographic separation led to the isolation of 6-Deoxybostrycoidin. The structure was determined from the interpretation of spectroscopic data (UV, MS, and NMR). 6-Deoxybostrycoidin exhibited in vitro activity against Bacillus subtilis, Staphylococcus aureus and Flexibacter maritimus with minimal inhibitory concentrations of 25, 12.5 and 12.5 μg/ml respectively.

Keywords: marine fungi, fish pathogenic bacteria, microorganism, medicine

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Authors: Abhichet Nobhiwong, Jiraporn Rojtinnakorn, Udomluk Sompong

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Six probiotic strains isolated from Chiang Mai and Chiang Rai province, Thailand; CR1-2, CM3-4, CM5-2, CR7-8, CM10-5 and CM10-8 were used to study their morphology and inhibition activity on three pathogenic bacteria; Aeromonas sp., Streptococcus sp. and Flavobacterium sp. that isolated from infected Nile tilapia. The agar well diffusion technique was applied for 24 and 48 hours incubation. Interestingly, some probiotics showed good inhibition activity both 24 and 48 hours on each 3 bacterial pathogens. The capable inhibiting Aeromonas sp. were CR1-2 and CR5-2 with inhibition diameters of 13.0 mm and 11.2 mm, respectively. For Streptococcus sp., effective probiotics were CR10-2 with inhibition diameters of 10.7 mm. Whereas for Flavobacterium sp., effective probiotics were CR5-2 with inhibition diameter of 9.7 mm. It can be concluded that these probiotics have potentiality to develop as the pathogens biocontrol products. These will be support for safety and organic aquaculture that which the most worthy for people health.

Keywords: probiotics, Aeromanas sp., Streptococcus sp., Flavobacterium sp.

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Authors: Jae Hong Kim, Sang Cheol Um, Jong Kook Lee

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Strong and biocompatible wollastonite (CaSiO3) was fabricated by pressureless sintering at temperature range of 1250~ 1300 ℃ and phase transition of to β-wollastonite with an addition of MgSiO3. Starting pure α-wollastonite powder were prepared by solid state reaction, and MgSiO3 powder was added to α-wollastonite powder to induce the phase transition α to β-wollastonite over 1250℃. Sintered wollastonite samples at 1250℃ with 5 and 10 wt% MgSiO3 were α+β phase and β phase respectively, and showed higher densification rate than that of α or β-wollastonite, which are almost the same as the theoretical density. Hardness and Young’s modulus of sintered wollastonite were dependent on the apparent density and the amount of β-wollastonite. Young’s modulus (78GPa) of β-wollastonite added 10 wt% MgSiO3 was almost double time of sintered α-wollastonite. From the in-vitro test, biphasic (α+β) wollastonite with 5wt% MgSiO3 addition had good bioactivity in simulated body fluid solution.

Keywords: β-wollastonite, high density, MgSiO3, phase transition

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Authors: Mohammad Hossein Pazandeh, Monir Doudi, Sona Rostampour Yasouri

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—Results The prudent administration of antibiotics aims to avoid the side effects and the microbes' resistance to antibiotics. An approach developing methods of local administration of antibiotics is especially required for localized infections caused by bacterial colonization of medical devices or implant materials. Among the wide variety of materials used as drug delivery systems, bioactive glasses (BG) have large utilization in regenerative medicine . firstly, the production of bioactive glass/nickel oxide/tin dioxide nanocomposite using sol-gel method, and then, the controlled release of imipenem from the double metal oxide/bioactive glass nanocomposite, and finally, the investigation of the antibacterial property of the nanocomposite. against a number of implant-related infectious agents. In this study, BG/SnO2 and BG/NiO single systema with different metal oxide present and BG/NiO/SnO2 nanocomposites were synthesized by sol-gel as drug carriers for tetracycline and imepinem. These two antibiotics were widely used for osteomyelitis because of its favorable penetration and bactericidal effect on all the probable osteomyelitis pathogens. The antibacterial activity of synthesized samples were evaluated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa as bacteria model using disk diffusion method. The BG modification using metal oxides results to antibacterial property of samples containing metal oxide with highest efficiency for nancomposite. bioactivity of all samples was assessed by determining the surface morphology, structural and composition changes using scanning electron microscopy (SEM), FTIR and X-ray diffraction (XRD) spectroscopy, respectively, after soaking in simulated body fluid (SBF) for 28 days. The hydroxyapatite formation was clearly observed as a bioactivity measurement. Then, BG nanocomposite sample was loaded using two antibiotics, separately and their release profiles were studied. The BG nancomposite sample was shown the slow and continuous drug releasing for a period of 72 hours which is desirable for a drug delivery system. The loaded antibiotic nanocomposite sample retaining antibacterial property and showing inactivation effect against bacteria under test. The modified bioactive glass forming hydroxyapatite with controlled release drug and effective against bacterial infections can be introduced as scaffolds for bone implants after clinical trials for biomedical applications . Considering the formation of biofilm by infectious bacteria after sticking on the surfaces of implants, medical devices, etc. Also, considering the complications of traditional methods, solving the problems caused by the above-mentioned microorganisms in technical and biomedical industries was one of the necessities of this research.

Keywords: antibacterial, bioglass, drug delivery system, sol- gel

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Authors: Abdu Zakari, Sai’d Jibril, Adoum A. Omar

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The leaves of Combretum molle was selected on the basis of its uses in folk medicine as insecticides. The leave extracts of Combretum molle was tested against the larvae of Artemia salina, i.e. Brine Shrimp Lethality Test (BST), Culex quinquefasciatus Say (Filaria disease vector) i.e. Larvicidal Test, using crude ethanol, n-hexane, chloroform, ethyl acetate, and methanol extracts. The methanolic extract proved to be the most effective in inducing complete lethality at minimum doses both in the BST and the Larvicidal activity test. The LC50¬ values obtained are 24.85 µg/ml and 0.4µg/ml respectively. The bioactivity-guided column chromatography afforded the pure compound ACM–3. ACM-3 was not active in the BST with LC50 value >1000µg/ml, but was active in the Larvicidal activity test with LC50 value 4.0µg/ml. ACM-3 was proposed to have the structure I, (Cholesta-β-D-Glucopyranoside).

Keywords: toxicity, larvicidal, Combretum molle, Artemia salina, Culex quinquefasciatus Say.

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Authors: Raquel C. A. G. Mota, Livia R. Menezes, Emerson O. da Silva

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Both silver nanoparticles and titanium dioxide have been extensively used in tissue engineering since they’ve been approved by the Food and Drug Administration (FDA), and present a bactericide effect when added to a polymeric matrix. In this work, the focus is on fabricating binary systems with both nanoparticles so that the synergistic effect can be investigated. The systems were tested by Nuclear Magnetic Resonance (NMR), Thermogravimetric Analysis (TGA), Fourier-Transformed Infrared (FTIR), and Differential Scanning Calorimetry (DSC), and X-ray Diffraction (XRD), and had both their bioactivity and bactericide effect tested. The binary systems presented different properties than the individual systems, enhancing both the thermal and biological properties as was to be expected. The crystallinity was also affected, as indicated by the finding of the DSC and XDR techniques, and the NMR showed a good dispersion of both nanoparticles in the polymer matrix. These findings indicate the potential of combining TiO₂ and silver nanoparticles in biomedicine.

Keywords: metallic nanoparticles, nanotechnology, polymer nanocomposites, polymer science

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Authors: Kanchan Maji, Debasmita Pani, Sudip Dasgupta

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Calcium phosphate cement (CPC) due to its high bioactivity and optimum bioresorbability shows excellent bone regeneration capability. Despite it has limited applications as bone implant due to its macro-porous microstructure causing its poor mechanical strength. The reinforcement of apatitic CPCs with biocompatible fibre glass phase is an attractive area of research to improve its mechanical strength. Here we study the setting behaviour of Si-doped and un-doped alpha tri-calcium phosphate (α-TCP) based CPC and its reinforcement with the addition of E-glass fibre. Alpha tri-calcium phosphate powders were prepared by solid state sintering of CaCO3, CaHPO4 and tetra ethyl ortho silicate (TEOS) was used as silicon source to synthesise Si doped α-TCP powders. Alpha tri-calcium phosphate based CPC hydrolyzes to form hydroxyapatite (HA) crystals having excellent osteoconductivity and bone-replacement capability thus self-hardens through the entanglement of HA crystals. Setting time, phase composition, hydrolysis conversion rate, microstructure, and diametral tensile strength (DTS) of un-doped CPC and Si-doped CPC were studied and compared. Both initial and final setting time of the developed cement was delayed because of Si addition. Crystalline phases of HA (JCPDS 9-432), α-TCP (JCPDS 29-359) and β-TCP (JCPDS 9-169) were detected in the X-ray diffraction (XRD) pattern after immersion of CPC in simulated body fluid (SBF) for 0 hours to 10 days. The intensities of the α-TCP peaks of (201) and (161) at 2θ of 22.2°and 24.1° decreased when the time of immersion of CPC in SBF increased from 0 hours to 10 days, due to its transformation into HA. As Si incorporation in the crystal lattice stabilised the TCP phase, Si doped CPC showed a little slower rate of conversion into HA phase as compared to un-doped CPC. The SEM image of the microstructure of hardened CPC showed lower grain size of HA in un-doped CPC because of premature setting and faster hydrolysis of un-doped CPC in SBF as compared that in Si-doped CPC. Premature setting caused generation of micro and macro porosity in un-doped CPC structure which resulted in its lower mechanical strength as compared to that in Si-doped CPC. This lower porosity and greater compactness in the microstructure attributes to greater DTS values observed in Si-doped CPC. E-glass fibres of the average diameter of 12 μm were cut into approximately 1 mm in length and immersed in SBF to deposit carbonated apatite on its surface. This was performed to promote HA crystal growth and entanglement along the fibre surface to promote stronger interface between dispersed E-glass fibre and CPC matrix. It was found that addition of 10 wt% of E-glass fibre into Si-doped α-TCP increased the average DTS of CPC from 8 MPa to 15 MPa as the fibres could resist the propagation of crack by deflecting the crack tip. Our study shows that biocompatible E-glass fibre in optimum proportion in CPC matrix can enhance the mechanical strength of CPC without affecting its bioactivity.

Keywords: Calcium phosphate cement, biocompatibility, e-glass fibre, diametral tensile strength

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Authors: Zhihao Peng, Qiong Zhang, Xiyao Li, Yongzhen Peng

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Nowadays, conventional nitrogen removal process (nitrification and denitrification) was adopted in most wastewater treatment plants, but many problems have occurred, such as: high aeration energy consumption, extra carbon sources dosage and high sludge treatment costs. The emergence of anammox has bring about the great revolution to the nitrogen removal technology, and only the ammonia and nitrite were required to remove nitrogen autotrophically, no demand for aeration and sludge treatment. However, there existed many challenges in anammox applications: difficulty of biomass retention, insufficiency of nitrite substrate, damage from complex organic etc. Much effort was put into the research in overcoming the above challenges, and the payment was rewarded. It was also imperative to establish an innovative process that can settle the above problems synchronously, after all any obstacle above mentioned can cause the collapse of anammox system. Therefore, in this study, a two-stage process was established that the sequencing batch reactor (SBR) and upflow anaerobic sludge blanket (UASB) were used in the pre-stage and post-stage, respectively. The domestic wastewater entered into the SBR first and went through anaerobic/aerobic/anoxic (An/O/A) mode, and the draining at the aerobic end of SBR was mixed with domestic wastewater, the mixture then entering to the UASB. In the long term, organic and nitrogen removal performance was evaluated. All along the operation, most COD was removed in pre-stage (COD removal efficiency > 64.1%), including some macromolecular organic matter, like: tryptophan, tyrosinase and fulvic acid, which could weaken the damage of organic matter to anammox. And the An/O/A operating mode of SBR was beneficial to the achievement and maintenance of partial nitrification (PN). Hence, sufficient and steady nitrite supply was another favorable condition to anammox enhancement. Besides, the flexible mixing ratio helped to gain a substrate ratio appropriate to anammox (1.32-1.46), which further enhance the anammox. Further, the UASB was used and gas recirculation strategy was adopted in the post-stage, aiming to achieve granulation by the selection pressure. As expected, the granules formed rapidly during 38 days, which increased from 153.3 to 354.3 μm. Based on bioactivity and gene measurement, the anammox metabolism and abundance level rose evidently, by 2.35 mgN/gVss·h and 5.3 x109. The anammox bacteria mainly distributed in the large granules (>1000 μm), while the biomass in the flocs (<200 μm) and microgranules (200-500 μm) barely displayed anammox bioactivity. Enhanced anammox promoted the advanced autotrophic nitrogen removal, which increased from 71.9% to 93.4%, even when the temperature was only 12.9 ℃. Therefore, it was feasible to enhance anammox in the multiple favorable conditions created, and the strategy extended the application of anammox to the full-scale mainstream, enhanced the understanding of anammox in the aspects of culturing conditions.

Keywords: anammox, granules, nitrite accumulation, nitrogen removal efficiency

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Authors: Alaa M. M. Badr Eldin, Marwa I. Ezzat, Mohammed S. Sedeek, Manal S. Afifi, Omar M. Sabry

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Cytotoxic activity of the total methanol extract against breast cancer cell line MCF-7 was amazing IC50 at 54 ug/ml. 130 polyphenolic compounds were tentatively identified in pomegranate peel (Punica granatum L.) methanol extract using HPLC-MS/MS technique. The antiestrogenic activity of the polyphenolic constituents found in pomegranate extract was confirmed experimentally in-vitro and by the in-silico molecular docking using gallagic acid, ellagic acid, and Punicalagin as these are considered model compounds confirmed in pomegranate peel extract. The methanolic extract was found to suppress ER, TGF-β, and NF-kB in-vitro gene expression strongly, and that was verified by qPCR and Western Blot gel electrophoresis techniques.

Keywords: HPLC-MS/MS, pomegranate, breast cancer, ovarian cancer, ER, TGF-β, NF-kB

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Authors: M. H. Pazandeh, M. Doudi, S. Barahimi, L. Rahimzadeh Torabi

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The use of antibiotics is essential in reducing the occurrence of adverse effects and inhibiting the emergence of antibiotic resistance in microbial populations. The necessity for a novel methodology concerning local administration of antibiotics has arisen, with particular focus on dealing with localized infections prompted by bacterial colonization of medical devices or implant materials. Bioactive glasses (BG) are extensively employed in the field of regenerative medicine, encompassing a diverse range of materials utilized for drug delivery systems. In the present investigation, various drug carriers for imipenem and tetracycline, namely single systems BG/SnO2, BG/NiO with varying proportions of metal oxide, and nanocomposite BG/SnO2/NiO, were synthesized through the sol-gel technique. The antibacterial efficacy of the synthesized samples was assessed through the utilization of the disk diffusion method with the aim of neutralizing Staphylococcus aureus as the bacterial model. The current study involved the examination of the bioactivity of two samples, namely BG10SnO2/10NiO and BG20SnO2, which were chosen based on their heightened bacterial inactivation properties. This evaluation entailed the employment of two techniques: the measurement of the pH of simulated body fluid (SBF) solution and the analysis of the sample tablets through X-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transform infrared (FTIR) spectroscopy. The sample tablets were submerged in SBF for varying durations of 7, 14, and 28 days. The bioactivity of the composite bioactive glass sample was assessed through characterization of alterations in its surface morphology, structure, and chemical composition. This evaluation was performed using scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, and X-ray diffraction spectroscopy. Subsequently, the sample was immersed in simulated liquids to simulate its behavior in biological environments. The specific body fat percentage (SBF) was assessed over a 28-day period. The confirmation of the formation of a hydroxyapatite surface layer serves as a distinct indicator of bioactivity. The infusion of antibiotics into the composite bioactive glass specimen was done separately, and then the release kinetics of tetracycline and imipenem were tested in simulated body fluid (SBF). Antimicrobial effectiveness against various bacterial strains have been proven in numerous instances using both melt and sol-gel techniques to create multiple bioactive glass compositions. An elevated concentration of calcium ions within a solution has been observed to cause an increase in the pH level. In aqueous suspensions, bioactive glass particles manifest a significant antimicrobial impact. The composite bioactive glass specimen exhibits a gradual and uninterrupted release, which is highly desirable for a drug delivery system over a span of 72 hours. The reduction in absorption, which signals the loss of a portion of the antibiotic during the loading process from the initial phosphate-buffered saline solution, indicates the successful bonding of the two antibiotics to the surfaces of the bioactive glass samples. The sample denoted as BG/10SnO2/10NiO exhibits a higher loading of particles compared to the sample designated as BG/20SnO2 in the context of bioactive glass. The enriched sample demonstrates a heightened bactericidal impact on the bacteria under investigation while concurrently preserving its antibacterial characteristics. Tailored bioactive glass that incorporates hydroxyapatite, with a regulated and efficient release of drugs targeting bacterial infections, holds promise as a potential framework for bone implant scaffolds following rigorous clinical evaluation, thereby establishing potential future biomedical uses. During the modification process, the introduction of metal oxides into bioactive glass resulted in improved antibacterial characteristics, particularly in the composite bioactive glass sample that displayed the highest level of efficiency.

Keywords: antibacterial, bioactive glasses, implant infections, multi drug resistant

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Authors: Paweł Stączek, Tomasz Plech, Aleksandra Strzelczyk, Katarzyna Dzitko, Monika Wujec, Edyta Kuśmierz, Piotr Paneth, Agata Paneth

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In this contribution, we will describe the inhibitory potency of nine thiosemicarbazide derivatives against bacterial type IIA topoisomerases, their antibacterial profile, and molecular modeling evaluation. We have found that one of the tested compounds, 4-benzoyl-1-(2-methyl-furan-3-ylcarbonyl) thiosemicarbazide, remarkably inhibits the activity of S. aureus DNA gyrase with the IC50 below 5 μM. Besides, this compound displays antibacterial activity on Staphylococcus spp. and E. faecalis at non-cytotoxic concentrations in mammalian cells, with minimal inhibitory concentrations (MICs) values at 25 μg/mL. Based on the enzymatic and molecular modeling studies we propose two factors, i.e. geometry of molecule and hydrophobic/hydrophilic balance as important molecular properties for developing thiosemicarbazide derivatives as potent Staphylococcus aureus DNA gyrase inhibitors.

Keywords: bioactivity, drug design, topoisomerase, molecular modeling

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Authors: Raja Arunprasath, P. Gajalakshmi

Abstract:

Antimycobacterial activity of C. roseus rosea and piperine was evaluated against ofloxacin resistant M. tuberculosis. Among the 68 suspected sputum samples, 32 were AFB positive belongs to age group of 40-50years. Susceptibility of M. tuberculosis was evaluated against ofloxacin and streptomycin by colorimetric assay. Of these 32 positive samples, 20 isolates were resistant to ofloxacin, 12 were resistant to Streptomycin and none of them were found to be multidrug resistant. The sensitivity pattern of ofloxacin resistant M. tuberculosis against two tested plant extracts showed potent tubercular activity. Antimycobacterial activity of C. roseus was 22 + 2.21mm and piperine was found to be 20 + 1.08 mm. The percentage of relative inhibitory zone of C. roseus was 133 % and piperine was found to be 111 %. The MIC of C. roseus and piperine was found at 50 µg/ml. Based on the FICI value 0.37 confirms that both the tested phytochemicals were synergistically active against M. tuberculosis. The MIC of ofloxacin was reduced from 8 mg to 2 mg/l in the presence of piperine but not by C. roseus. This is the first report on Synergistic bioactivity of C. roseus rosea and piperine fractionation leads development of novel antimycobacterial prophylaxis in future.

Keywords: C. roseus, ofloxacin, piperine, synergistic

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Authors: Sani Abubakar, Oumar Al-Mubarak Adoum

Abstract:

The leaves extracts of Cassia sieberiana D. C. were screened for antimicrobial bioassay against Staphylococcus aureus, Salmonella typhi, and Escherichia coli and cytotoxicity using Brine Shrimp Test (BST). The crude ethanol extract, Chloroform soluble fraction, aqueous soluble fraction, ethyl acetate soluble fraction, methanol soluble fraction, and n-hexane soluble fraction were tested against antimicrobial and cytotoxicity. The Ethyl acetate fraction obtained proved to be most active in inducing complete lethality at minimum doses in BST and also active on Salmonella typhi. The bioactivity result was used to guide the column chromatography, which led to the isolation of pure compound CSB-8, which was found active in the BST with an LC₅₀ value of 34(722-182)µg/ml and showed remarkable activity on Salmonella typhi (zone of inhibition 25mm) at 10,000µg/ml. The ¹H-NMR, ¹³C NMR, FTIR, and GC-MS spectra of the compound suggested the proposed structure to be 2-pentadecanone.

Keywords: antimicrobial bioassay, cytotoxicity, column chromatagraphy, Cassia sieberiana D.C.

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Authors: Aura Maria Lopera Echavarria, Angela Maria Lema Perez, Daniela Medrano David, Pedronel Araque Marin, Marta Elena Londoño Lopez

Abstract:

Bone regeneration is the process by which the formation of new bone is stimulated. Bone fractures can originate at any time due to trauma, infections, tumors, congenital malformations or skeletal diseases. Currently there are different strategies to treat bone defects that in some cases, regeneration does not occur on its own. That is why they are treated with bone substitutes, which provide a necessary environment for the cells to synthesize new bone. The Demineralized Bone Matrix (DBM) is widely used as a bone implant due to its good properties, such as osteoinduction and bioactivity. However, the use of DBM is limited, because its presentation is powder, which is difficult to implant with precision and is susceptible to migrating to other sites through blood flow. That is why the DBM is commonly incorporated into a variety of vehicles or carriers. The objective of this project is to evaluate the bioactive and confinement properties of a bone substitute based on demineralized bone matrix (DBM). Also, structural and morphological properties were evaluated. Bone substitute was obtained from EIA Biomaterials Laboratory of EIA University and the DBM was facilitated by Tissue Bank Foundation. Morphological and structural properties were evaluated by scanning electron microscopy (SEM), X-ray diffraction (DRX) and Fourier transform infrared spectroscopy with total attenuated reflection (FTIR-ATR). Water absorption capacity and degradation were also evaluated during three months. The cytotoxicity was evaluated by the MTT test. The bioactivity of the bone substitute was evaluated through immersion of the samples in simulated body fluid during four weeks. Confinement tests were performed on tibial fragments of a human donor with bone defects of determined size, to ensure that the substitute remains in the defect despite the continuous flow of fluid. According of the knowledge of the authors, the methodology for evaluating samples in a confined environment has not been evaluated before in real human bones. The morphology of the samples showed irregular surface and presented some porosity. DRX confirmed a semi-crystalline structure. The FTIR-ATR determined the organic and inorganic phase of the sample. The degradation and absorption measurements stablished a loss of 3% and 150% in one month respectively. The MTT showed that the system is not cytotoxic. Apatite clusters formed from the first week were visualized by SEM and confirmed by EDS. These calcium phosphates are necessary to stimulate bone regeneration and thanks to the porosity of the developed material, osteinduction and osteoconduction are possible. The results of the in vitro evaluation of the confinement of the material showed that the migration of the bone filling to other sites is negligible, although the samples were subjected to the passage of simulated body fluid. The bone substitute, putty type, showed stability, is bioactive, non-cytotoxic and has handling properties for specialists at the time of implantation. The obtained system allows to maintain the osteoinductive properties of DBM and it can fill completely fractures in any way; however, it does not provide a structural support, that is, it should only be used to treat fractures without requiring a mechanical load.

Keywords: bone regeneration, cytotoxicity, demineralized bone matrix, hydrogel

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Authors: Mohammad Shakir, Reshma Jolly, Mohammad Shoeb Khan, Noor-E-Iram

Abstract:

A nanocomposite system incorporating dextrin into nano-hydroxyapatite/chitin matrix (n-HA/DX/CT) has been successfully synthesized via co-precipitation route at room temperature for the application in bone tissue engineering by investigating biocompatibility, cytotoxicity and mechanical properties. The FTIR spectra of n-HA/DX/CT nanocomposite indicated a considerable intermolecular interaction between the various components of the system. The results of XRD, TEM and TGA/DTA revealed that the crystallinity, size and thermal stability of the n-HA/DX/CT scaffold has decreased and increased respectively. The result of SEM image of the n-HA/DX/CT scaffold indicated that the incorporation of dextrin affected the surface morphology while considerable in-vitro bioactivity has been observed in n-HA/DX/CT based on SBF study, referring a step towards possibility of making direct bond to living bone if implanted. Moreover, MTT assay suggested the non-toxic nature of n-HA/DX/CT to murine fibroblast L929 cells. The swelling study of n-HA/DX/CT scaffold indicated the low swelling rate for n-HADX/CT. All these results have paved the way for n-HA/DX/CT to be used as a competent material for bone tissue engineering.

Keywords: autograft, chitin, dextrin, nanocomposite

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Authors: Jong Kook Lee, Sangcheol Eum, Jaehong Kim

Abstract:

Wollastonite/hydroxyapatite composite coatings on zirconia were obtained by room temperature spray process. Wollastonite powder was synthesized by solid-state reaction between calcite and silica powder. Hydroxyapatite powder was prepared from bovine bone by the calcination at 1200oC 1h. From two starting raw powders, three kinds of powder mixture were obtained by the ball milling for 24h. By using these powders, wollastonite/hydroxyapatite coatings were fabricated on zirconia substrates by a room temperature spray process, and their microstructure and biological behavior were investigated and compared with pure wollastonite and hydroxyapatite coatings. Wollastonite/hydroxyapatite coatings on zirconia substrates were homogeneously formed in microstructure and had a nanoscaled grain size. The phase composition of the resultant wollastonite/hydroxyapatite coatings was similar to that of the starting powders, however, the grain size of the wollastonite or hydroxyapatite particles was reduced to about 100 nm due to their formation by particle impaction and fracture. The wollastonite/hydroxyapatite coating layer exhibited bioactivity in a stimulated body fluid and forming ability of new hydroxyapatite precipitates of 25 nm during in vitro test in SBF solution, which was enhanced by the increasing wollastonite content.

Keywords: wollastonite, hydroxyapatite composite coatings, room temperature spay process, zirconia

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Authors: Paul Awolade, Parvesh Singh

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The global threat of pathogenic resistance to available therapeutic agents has become a menace to clinical practice, public health and man’s existence inconsequential. This has therefore led to an exigency in the development of new molecular scaffolds with profound activity profiles. In this vein, a versatile synthetic tool for accessing new molecules by incorporating two or more pharmacophores into a single entity with the unique ability to be recognized by multiple receptors hence leading to an improved bioactivity, known as molecular hybridization, has been explored with tremendous success. Accordingly, aware of the similarity in pharmacological activity spectrum of quinoline and 1,2,3-triazole pharmacophores such as; anti-Alzheimer, anticancer, anti-HIV, antimalarial and antimicrobial to mention but a few, the present study sets out to synthesize hybrids of quinoline and 1,2,3-triazole. The hybrids were accessed via click chemistry using copper catalysed azide-alkyne 1,3-dipolar cycloaddition reaction. All synthesized compounds were evaluated for their pharmaco-potential in an antimicrobial assay out of which the 3-OH derivative emerged as the most active with MIC value of 4 μg/mL against Cryptococcus neoformans; a value superior to standard Fluconazole and comparable to Amphotericin B. Structures of synthesized hybrids were elucidated using appropriate spectroscopic techniques (1H, 13C and 2D NMR, FT-IR and HRMS).

Keywords: bioisostere, click chemistry, molecular hybridization, quinoline, 1, 2, 3-triazole

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Authors: D. Baranova, E. Neborak

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Polyphenols, particularly flavonoids like quercetin, fisetin, and kaempferol, have gained significant attention in nutrition due to their antioxidant, senolytic, and anti-inflammatory properties. These compounds are commonly found in various plant-based foods and are represented by diverse subclasses, each with unique health benefits. Understanding their absorption, metabolism, and bioactivity within the human body is crucial for unlocking their full potential. Quercetin, for instance, exists in multiple forms, impacting its solubility and absorption in the intestine. Its intake, often derived from sources like apples, is affected by cooking methods, with medium heat retaining its potency. Fisetin, also present in fruits and vegetables, demonstrates neuroprotective qualities and stability under varied conditions compared to quercetin. Similarly, kaempferol, found in fruits and vegetables, displays antioxidative effects but is influenced by cooking techniques, with specific methods preserving its polyphenolic content better. Overall, these polyphenols offer promising health benefits, yet their optimal dosage and specific dietary recommendations warrant further research to harness their full nutritional potential.

Keywords: polyphenols, flavonoids, absorption, quercetin, kaempferol, fisetin, senolytics, absorption, cooking method

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Authors: Maryam Ehsanpou, Majid Afkhami, Flora Mohammadizadeh, Amirhoushang Bahri, Rastin Afkhami

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Within the frame of a biodiversity and bioactivity study of marine macro organisms from the Persian Gulf, three hard coral species extracts were investigated for cytotoxic, antibacterial and antifungal activities against five human pathogenic microorganisms. All concentrations of extracts from three hard corals showed no antifungal activity towards the tested strains. In antibacterial assays, the hard coral extracts showed significant activity solely against Staphylococcus aureus with MICs ranging from 3 to 9 μg/ml. The highest antibacterial activity was found in the aqueous methanol extract of Porites compressa with an inhibition zone of 22 mm against Staphylococcus aureus at 18 μg/ml extract concentration. Methanol extracts from Porites harrisoi and Porites compressa exhibited only weak cytotoxic activities. It is important for future research to concentrate on finding the mechanisms employed by corals to defend themselves against invasion, the mechanism of infections and the type of chemical compounds in coral extracts that inhibit antibacterial growth or proliferation in underexplored areas such as the Persian Gulf.

Keywords: antibacterial, antifungal, cytotoxic, hard corals, Persian Gulf

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Authors: Jiraporn Rojtinnakorn, Mallika Supa-Aksorn, Sudaporn Tongsiri, Prachaub Chaibu

Abstract:

Marble goby is one of high demand consuming in Southeast Asia. However, the product was from riparian fisheries because of low yield in aquaculture, especially in nursery stage. Therefore, we studied for herb supplement in pellet feed of marble goby fingering. Garlic, a common herb and illustrated novel pharmaceutical and medical effectiveness, was considered. Garlic extracts with water (DW), 50% EtOH (50E), 95% EtOH (95E) and diethyl ether (DE) were subjected for feed additive to induce immune response in marble goby fingering for 0 (control), 0.3, 0.5, 1.0, 3.0 and 5.0 % (w/w). After seven days of feeding, blood was collected for analysis of blood composition; i.e. haematocrit (HCT), red blood cells (RBC), white blood cells (WBC) and humoral immune responses; i.e. lysozyme activity (Lys). It was resulted that values of HCT, WBC and Lys in all garlic fed group were significantly different from control (p < 0.05). For HCT, the highest values belonged to 5% DW and 0.5% 95E. For WBC and Lys, the highest values were 5% DW. For RBC, there was not obviously significant (p < 0.05). There were only 3 groups; 0.5% 95E, 1% and 5% DW, showed distinct statistical significance from the other groups. It was concluded that garlic extracts showed satisfy bioactivity to enhancing innate immune response in marble goby fingering. This result will be valuable for specific feed formula of marble goby nursery.

Keywords: garlic extract, innate immune, marble goby, Oxyeleotris marmoratus

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Authors: SeyedMohammad Hosseini, Amirhossein Moghanian

Abstract:

The main purpose of this research was to design a biomimetic system by freeze-drying method for evaluating the effect of adding 5 and 10 mol. % of zinc (Zn)in 58S bioactive glass and gelatin (5ZnBG/G and 10ZnBG/G) in terms of structural and biological changes. The structural analyses of samples were performed by X-Ray Diffraction (XRD), scanning electron microscopy (SEM), and Fourier-transform infrared spectroscopy (FTIR). Also, 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT) and alkaline phosphate (ALP) activity test were carried out for investigation of MC3T3-E1cell behaviors. The SEM results demonstrated the spherical shape of the formed hydroxyapatite (HA) phases, and also HA characteristic peaks were detected by X-ray diffraction spectroscopy (XRD)after 3 days of immersion in the simulated body fluid (SBF) solution. Meanwhile, FTIR spectra proved that the intensity of P–O peaks for 5ZnBG/G was more than 10ZnBG/G and control samples. Moreover, the results of alkaline phosphatase activity (ALP) test illustrated that the optimal amount of Zn (5ZnBG/G) caused a considerable enhancement in bone cell growth. Taken together, the scaffold with 5 mol.% Zn was introduced as an optimal sample because of its higher biocompatibility, in vitro bioactivity, and growth of MC3T3-E1cellsin in comparison with other samples in bone tissue engineering.

Keywords: scaffold, gelatin, modified bioactive glass, alp, bone tissue engineering

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Authors: Funmilola O. Omoya

Abstract:

Muscid flies are known to be vectors of disease agents and species that annoy humans and domesticated animals. An example of these flies is Musca domestica (house fly) whose adult and immature stages occur in a variety of filthy organic substances including household garbage and animal manures. They contribute to microbial contamination of foods. It is therefore imperative to control these flies as a result of their role in Public health. The second and third instars of Musca domestica (Linn) were infected with varying cell loads of Bacillus subtilis in vitro for a period of 48 hours to evaluate its larvicidal activities. Mortality of the larvae increased with incubation period after treatment with the varying cell loads. Investigation revealed that the second instars larvae were more susceptible to treatment than the third instars treatments. Values obtained from the third instar group were significantly different (P0.05) from those obtained from the second instars group in all the treatments. Lethal concentration (LC50) at 24 hours for 2nd instars was 2.35 while LC50 at 48 hours was 4.31.This study revealed that Bacillus subtilis possess good larvicidal potential for use in the control of Musca domestica in poultry farms.

Keywords: Bacillus subtilis, Musca domestica, larvicidal activities, poultry farms

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Authors: Indu Lata Kanwar, Preeti K. Suresh

Abstract:

The aim of this study is to fabricate hydroxyapatite based nanocomposites for local drug delivery in periodontal pockets. Hydroxyapatite is chemically similar to the mineral component of bones and hard tissues in mammals. Synthetic biocompatibility and bioactivity with human teeth and bone, making it very attractive for biomedical applications. Nanocomposite is a multiphase solid material where one of the phases has one, two or three dimensions of less than 100 nanometres (nm), or structures having nano­scale repeat distances between the different phases that make up the material. Nanostructured calcium phosphate materials play an important role in the formation of hard tissues in nature. It is reported that calcium phosphates materials in nano-size can mimic the dimensions of constituent components of calcified tissues. Nano-sized materials offer improved performances compared with conventional materials due to their large surface-to-volume ratios. The specific biological properties of the nanocomposites, as well as their interaction with cells, include the use of bioactive molecules. The approach of periodontal tissue engineering is considered promising to restore bone defect through the use of engineered materials with the aim that they will prohibit the invasion of fibrous connective tissue and help repair the function during bone regeneration.

Keywords: bioactive, hydroxyapatite, nanocomposities, periondontal

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Authors: Muhammad Mustaqeem, Musa Kaleem Baloch, Irfan Ullah, Ammarah Luqman, Afshan Ahmad

Abstract:

Flurbiprofen is one of the most potent nonsteroidal anti-inflammatory drugs. It is widely used for relief of pain in patients suffering from rheumatic diseases, migraine, sore throat and primary dysmenorrhea. However, its aqueous solubility is very low and hinders the skin permeation. Thus, it is imperative to develop such a drug delivery systems which can improve its aqueous solubility and hence improve the skin permeation and therapeutic compliance. Microemulsions have been also proven to increase the cutaneous absorption of lipophilic drugs as compared to conventional vehicles. Micro-emulsion is thermodynamically stable emulsion that has the capacity to ‘hide/solubilize’ water-insoluble molecules within a continuous oil phase. Therefore, flurbiprofen was converted to Easters through chemical reactions with alcohols such as methanol, ethanol, propanol and butanol. The product was further treated with hydrazine to get hydrazide. The solubility of the parent drug Flurbiprofen and the products were solubilized in microemulsions formed using various surfactants like ionic, non-ionic and zwitterions. It has been concluded that the product was more soluble than the parent compound. The biological activities of these were also investigated. The outcome was very promising and the product was more active than the parent compound. It, therefore, concluded that in this way, we can not only enhance the solubility of the drug and increase its bioactivity, but also reduce the risk of stomach cancer.

Keywords: Flurbiprofen, microemulsion, surfactants, hyrazides

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