Polymer-based nanocomposites have attracted a lot of attention for amperometric biosensor development due to their general physical and chemical properties including biocompatibility, film-forming ability, stability and different functional groups that can be bonded with other biomolecues. In this study, poly-4-vinlyridine homopolymer (P4VP) and polylactic acid-block-poly(2-vinylpyridine) block copolymer (PLA-b-P2VP) were used to hybridize with gold precursors (Au3+) based on the association between the nitrogen of the pyridine group of P4VP or P2VP block with gold precursors. P4VP/Au3+ and PLA-b-P2VP/Au3+ nanocomposites were prepared with ratio of gold to P2VP or P4VP (10:1). The Au3+ in both polymers was reduced to gold nanoparticles (AuNPs) via in-situ approach by using hydrazine. Fourier transform infrared spectroscopy (FTIR), ultraviolet-visible spectroscopy (UV-vis), transmission electron microscopy (TEM) and cyclic voltammetry (CV) were used to characterize the structural, morphological and electrochemical properties of the nanocomposites. The peak currents of P4VP/AuNPs and PLA-b-P2VP/AuNPs nanocomposites modified electrode were 6.685 nA and 69.432 nA, respectively, which are much lower than bare electrode (205.019 nA) due to the non-conductivity of P4VP and PLA-b-P2VP. In order to improve the electron transfer capability of electrode, graphene oxide (GO) was blended and electrochemically reduced to obtain P4VP/AuNPs/rGO and PLA-b-P2VP/AuNPs/rGO nanocomposites. After immobilization of these two nanocomposites on electrode through drop casting method, the peak currents of P4VP/AuNPs/rGO and PLA-b-P2VP/AuNPs/rGO nanocomposites modified electrode were 871.172 nA and 663.947 nA, respectively, which are much higher than bare electrode (205.019 nA) and shown good capability to accelerate electron transfer. Based on these characterizations, P4VP/AuNPs/rGO has potential as the nanocomposite to modify the electrode for enzymatic biosensor development.
Catalytic hydrogenation of carbon dioxide (CO2) to methanol is an attractive way to recycle and utilize CO2. A series of Cu/ZnO/Al2O3/ZrO2 catalysts (CZAZ) containing different molar ratios of Cu/Zn were prepared by the co-precipitation method. The catalysts were characterized by temperature-programmed reduction (TPR), field emission scanning electron microscopy-energy dispersive x-ray analysis (FESEM-EDX) and X-ray diffraction (XRD). Higher surface area, SABET values (42.6-59.9 m2/g) were recorded at low (1) and high (5) Cu/Zn ratios with the minimum value of 35.71 m2/g was found for a Cu/Zn of 3. The reducibility of the metal oxides formed after calcination of catalyst samples was also affected due to change in metal-support interaction. At a reaction temperature of 443 K, total gas pressure of 3.0 MPa and 0.1 g/mL of the CZAZ catalyst, the selectivity to methanol decreased as the Cu/Zn molar ratio increased, and the maximum selectivity of 93.9 was achieved at Cu/Zn molar ratio of 0.33. With a reaction time of 3h, the best performing catalyst was CZAZ75 with Cu/Zn molar ratio of 5 giving methanol yield of 6.4%.
Breast cancer is the abnormal, uncontrollable proliferation of cells in the breast. Conventional treatment modalities like chemotherapy induce deteriorating side effects on healthy cells. Non-viral inorganic nanoparticles (NPs) confer exclusive characteristics, such as, stability, controllable shape and size, facile surface modification, and unique magnetic and optical properties which make them attractive drug carriers. Among them, carbonate apatite (CA) particles are pH-responsive in nature, enabling rapid intracellular drug release, but are typically heterogeneous with the tendency to self-aggregate. Here, we modified the nano-carrier by partially substituting Ca2+ with Mg2+ and Fe3+ into a basic lattice structure of CA, forming Fe/Mg-carbonate apatite (Fe/Mg-CA) NPs with the ability to mitigate self-aggregation, form unique protein corona in the presence of serum and efficiently deliver doxorubicin (DOX), an anti-cancer drug into breast cancer cells. Two formulations of Fe/Mg-CA NPs were generated by adding different concentrations of Fe3+ and Mg2+ along with a fixed amount of Ca2+ in bicarbonate buffered DMEM (Dulbecco's Modified Eagle's Medium), followed by 30 min incubation at 37 °C. Particles were characterized by turbidity analysis, z-average diameter and zeta potential measurement, optical microscopy, field emission scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FTIR), energy dispersive X-ray (EDX), flame atomic absorption spectroscopy (FAAS), pH dissolution, drug binding, cellular uptake, thiazolyl blue tetrazolium bromide (MTT) assay, stability analysis, and protein corona study by LCMS (Liquid chromatography-mass spectrometry). Both formulations of Fe/Mg-CA displayed mostly uniform nano-sized particles with less tendency to aggregate. The EDX and FAAS elemental analysis confirmed the weight (%) of Ca, Fe and Mg, along with their Ca/P ratio in the particles. A constant drug binding efficiency was noticed with 5 μM to 10 μM of initial DOX concentration. A pH dissolution study of Fe/Mg-CA NPs revealed the quick release of DOX in acidic pH. Enhancement of cytotoxicity for the chemotherapy drug was greater for Fe/Mg-CA NPs as compared to CA NPs, which could be explained by an increase in cellular internalization as a result of the small z-average diameter of the former. The protein corona study by LCMS demonstrated that Fe/Mg-CA NPs exhibited the highest affinity towards transport proteins without binding with opsonins. Biodistribution study was performed to study the effect of DOX-loaded Fe/Mg-CA NPs on the tissue distribution of DOX in Balb/c 4T1 tumor-bearing mice. Both formulations of Fe/Mg-CA NPs have significantly increased the accumulation of DOX in tumors. Interestingly, high Fe/Mg-CA NPs exhibited less off-target distribution compared to low Fe/Mg-CA NPs. Furthermore, the blood plasma analysis revealed prolonged blood circulation half-life of DOX-loaded low and high Fe/Mg-CA NPs compared to free DOX solution. Modifying CA NPs with Fe3+ and Mg2+, thereby, led to the generation of nano-sized particles with less tendency to aggregate, enhancing the drug binding efficiency, cellular uptake, and cytotoxicity without hampering drug release in acidic pH, while improving the circulation half-life and tumor accumulation of DOX. Therefore, Fe/Mg-CA which predominantly forms a transport protein-related protein corona could be a proficient carrier for therapeutic delivery in breast cancer.
Demand for medical implants is rising day by day as the world becomes the place for more diseased and older people. Accordingly, in this research, metallocene polyethylene (mPE), a commonly used polymer was treated with UV rays for improving its biocompatibility. Scanning electron microscopy (SEM) images confirmed the formation of crests and troughs, which depicts the improvement of surface roughness of mPE substrates caused by UV etching. Accordingly, the contact angle measurements revealed that the wettability of mPE-2.5 J/cm2 (68.09º) and mPE-5 J/cm2 (57.93º) samples were found to be increased compared to untreated mPE (86.84º) indicating better hydrophilicity. Further, the UV treated surface exhibited enhanced blood compatibility as determined in APTT (untreated mPE- 55.3 ± 2.5 s, mPE-2.5 J/cm2 - 76.7 ± 4.1 s and mPE-5 J/cm2 - 112.3 ± 2 s) and PT (untreated mPE - 24.7 ± 1.5 s, mPE- 2.5 J/cm2 - 34.3 ± 1.1 s and mPE-5 J/cm2 - 43 ± 2 s) assay. Moreover, the treated mPE-2.5 J/cm2 (4.88%) and mPE-5 J/cm2 (1.79%) showed decreased hemolytic percentage compared to untreated mPE (15.40%) indicating better safety to red blood cells. Interestingly, the changes in physicochemical properties of mPE are directly proportional to the dosage of the UV rays. UV modified mPE surfaces were found to be more compatible as identified through MTT assay, photomicrograph and SEM images of the seeded 3T3 cell population. Hence UV-modified surface of mPE may be successfully exploited for medical implants.
Ayurveda oil contains numerous source of biological constituents which plays an important role in reducing the pain relief caused during bone fracture. The aim of the study is to fabricate the polyurethane (PU) scaffold for bone tissue engineering added with ayurveda amla oil using electrospinning technique. Scanning Electron Microscopy (SEM) analysis showed that the fabricated nanocomposites showed reduced fiber diameter (758 ± 185.46 nm) than the pristine PU (890 ± 116.91 nm). Fourier Infrared Analysis (FTIR) revealed the existence of amla oil in the PU matrix by hydrogen bond formation. The contact angle results revealed the decreased wettability (116° ± 1.528) of the prepared nanocomposites compared to the pure PU (100° ± 0.5774). The incorporation of amla oil into the PU matrix improved the surface roughness. Further, the coagulation assay indicated that the addition of amla oil into PU delayed the blood clotting times and exhibited less toxic to red blood cells. Hence, the fabricated nanocomposites showed enhanced physicochemical and better blood compatibility parameters which may serve as a potential candidate for bone tissue engineering.
The aim of this study was to develop polyurethane (PU) wound dressing incorporated with cobalt nitrate using electrospinning technique. The morphology analysis revealed that the developed composites exhibited reduced fiber and pore diameter than the pristine PU. The electrospun membranes exhibited average porosity in the range of 67% - 71%. Energy-dispersive X-ray spectra (EDS) showed the presence of cobalt in the PU matrix. The interaction of cobalt nitrate with PU matrix was evident in Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). The contact angle results indicated the improved wettability of the prepared PU/cobalt nitrate composites (82° ± 2) than the pure PU (100° ± 1). The incorporation of cobalt nitrate into the PU matrix enhanced the surface roughness and mechanical strength as evident in the atomic force microscopy (AFM) and tensile test analysis. The blood compatibility assays revealed the anticoagulant nature of the prepared composites by displaying prolonged blood clotting time than the PU control. Further, the developed composite exhibited less toxicity nature as revealed in the hemolysis and cytotoxicity studies. It was observed that the PU wound dressing added with cobalt nitrate fibers exhibited enhanced physicochemical, better blood compatibility parameters and enhanced fibroblast proliferation rates which may serve as a potential candidate for wound dressings.
The climbing abilities of two bed bug species, Cimex lectularius L. and Cimex hemipterus (F.), were determined by evaluating their escape rates from smooth surface pitfall traps using four commercial bed bug monitors (Verifi Bed Bug Detector, ClimbUp Insect Interceptor, BlackOut Bed Bug Detector, and SenSci Volcano Bed Bug Detector). All detectors were used in the absence of lures or attractants. Unlike C. lectularius, adult C. hemipterus were able to escape from all traps. On the other hand, no or a low number nymphs of both species escaped, depending on the evaluated traps. Examination of the vertical friction force of adults of both species revealed a higher vertical friction force in C. hemipterus than in C. lectularius. Scanning electron microscope micrograph observation on the tibial pad of adult bed bugs of C. hemipterus showed the presence of a greater number of tenent hairs on the tibial pad than on that of adult C. lectularius. No tibial pad was found on the fourth and fifth instars of both species. Near the base of the hollow tenent hairs is a glandular epithelium that is better developed in adult C. hemipterus than in adult C. lectularius. This study highlights significant morphological differences between C. lectularius and C. hemipterus, which may have implications in the monitoring and management of bed bug infestations.
Bacterial mediated Silver nanoparticles is considered as an emerging Ecofriendly approach to eradicate human pathogens. This paper aims to provide the biological approach for the synthesis of silver nanoparticles from indigenously isolated bacteria. This study will be beneficial to control the nosocomial infections triggered by MRSA (Methicillin-resistant Staphylococcus aureus). The current study is the extracellular synthesis of silver nanoparticles by using the cell free filtrate of bacterial strains isolated from the soil. The optimization study was also carried out to obtain the maximum production of silver nanoparticles. Nanoparticles were confirmed and characterized by UV-Vis spectroscopy and Transmission Electron Microscopy (TEM) having the plasmon resonance peak between 420-450nm with 10-60nm in size range and most were spherical in shape. Synthesized silver nanoparticles showed a potential antibacterial activity against MRSA (Methicillin Resistant Staphylococcus aureus) in-vitro study. This is the green approach for the production of AgNPs, as there was no previous work done on the synthesis of silver nanoparticles by bacteria in this region of Southern Punjab, Pakistan and these nanoparticles can be used to treat nosocomial infection. These silver nanoparticles can be used in effective disease management as antimicrobial agent.
Percolation theory has been used with great interest in understanding the design and characterization of dosage forms. In this study, work has been carried out to investigate the behavior of binary mixture tablets containing excipients of similar and different deformation properties. The binary mixture tablets were prepared by direct compression using lactose, polyvinyl chloride (PVC), Eudragit RS 100, and microcrystalline cellulose (MCC). The application of percolation theory on the relationships between compactibility, Pmax, or compression susceptibility (compressibility), gamma, and mixture compositions reveals the presence of percolation thresholds even for mixtures of similar deformation properties. The results showed that all mixture compositions exhibited at least one discreet change in the slope, which was referred to as the percolation threshold. The PVC/Eudragit RS100 mixture compositions showed significant percolation threshold at 80% (w/w) PVC loading. Two percolation thresholds were observed from a series of binary mixtures containing similar plastic deformation materials (PVC/MCC). The percolation thresholds were determined at 20% (w/w) and 80% (w/w) PVC loading. These are areas where one of the components percolates throughout the system and the properties of the tablets are expected to experience a sudden change. Experimental results, however, showed that total disruption of the tablet physical properties at the specified percolation thresholds can be observed for PVC/lactose mixtures at 20-30% (w/w) loading while only minor changes in the tablets' strength for PVC/MCC or PVC/Eudragit RS 100 mixtures were observed.
It is generally accepted that the tablet elastic relaxation during compaction plays a vital role in undermining the final tablet mechanical integrity. One of the least investigated stages of the compaction process is the ejection stage.
The effects of a mixed infection of Mycoplasma gallinarum and Newcastle disease virus (F strain) on the tracheal epithelium of village chickens were investigated and observed by scanning electron microscopy. Day-old village chicks were vaccinated intranasally with F strain Newcastle disease virus and inoculated intratracheally on the same day with 10(8) colony forming units of M gallinarum. In another study the chicks were vaccinated and then infected with M gallinarum seven days later. The first group of chicks was euthanased three, seven, 10, 14 and 21 days after vaccination and infection and the vaccinated chicks were euthanased three, seven, 10 and 14 days after infection. In the chicks vaccinated and infected on the same day, major alterations to the tracheal epithelium were visible three days later. There were fewer ciliated cells and the borders of the non-ciliated cells were prominent. Several depressions had formed in the epithelial surface. At higher magnification, clumps of microvilli were visible on some of the non-ciliated cells. Seven days after vaccination and infection, the tracheal epithelium appeared normal, with an increase in the numbers of ciliated cells, although raised borders were observed on the non-ciliated cells in some areas. No clumping of microvilli or depressions in the epithelial surface were observed. In the chicks infected seven days after vaccination, the tracheal epithelium appeared normal with no visible changes on its surface.
The effects of exogenously introduced oestradiol-17 beta (E) and relaxin (RLX) on cervical extensibility and collagen organisation were tested in rats ovariectomised in late pregnancy. When the cervices were stretched in vitro by 1 mm increments, it was found that those from rats given E alone generated significantly higher tensions than those from control rats, while cervices from rats given both E and RLX had tensions similar to controls. Examination of cervical sections under the light microscope and ultra-thin sections under the electron microscope showed that the collagen fibres in the cervices from E-treated rats were highly organised, whereas those from animals given E+RLX and control animals were disorganised and dispersed. It was concluded that E decreased cervical extensibility, while RLX counteracted the effect of E to maintain a soft and easily extensible cervix.
Two experiments were conducted to evaluate the effect of formaldehyde vaporization of a hatcher on the tracheal epithelium of chick embryos, and on the production performance and behaviour of commercial broiler chicks. In experiment 1, chick embryos were exposed to 23.5 ppm of formaldehyde vapour during the last 3 days of incubation. Tracheal samples were taken at 0, 6, 30 and 54 h after exposure to formaldehyde and examined by scanning electron microscopy for pathological changes. Observable lesions included excessive accumulation of mucus, matted cilia, loss of cilia and sloughing of the epithelium. The lesions were more severe in chicks exposed for 54 h as compared to those exposed for 6 or 30 h. In experiment 2, 60 chicks that had been exposed to formaldehyde vapour as above and 60 control chicks were used to investigate the effect of formaldehyde fumigation on production performance and behaviour. Formaldehyde vaporization resulted in higher weekly (days 0-6 and 21-27) and total (days 0-41) feed intake and poorer weekly (days 0-6, 7-13, 21-27 and 28-34) and overall (days 0-41) feed conversion ratios. Body weight, mortality and behaviour (eating, drinking, sitting and standing activities) were not affected by formaldehyde fumigation.
The aim of this study was to prepare β-wollastonite using a green synthesis method (autoclaving technique) without organic solvents and to study its bioactivity. To prepare β-wollastonite, the precursor ratio of CaO:SiO₂ was set at 55:45. This mixture was autoclaved for 8 h and later sintered at 950 °C for 2 h. The chemical composition of the precursors was studied using X-ray fluorescence (XRF), in which rice husk ash consists of 89.5 wt % of SiO₂ in a cristobalite phase and calcined limestone contains 97.2 wt % of CaO. The X-ray diffraction (XRD) patterns after sintering showed that only β-wollastonite was detected as the single phase. To study its bioactivity and degradation properties, β-wollastonite samples were immersed in simulated body fluid (SBF) for various periods of time. Throughout the soaking period, the molar ratio of Ca/P obtained was in the range of 1.19 to 2.24, and the phase detected was amorphous calcium phosphate, which was confirmed by scanning electron microscope with energy dispersive X-ray analysis (SEM/EDX) and XRD. Fourier-transform infrared spectroscopy (FTIR) analysis indicated that the peaks of the calcium and phosphate ions increased when an amorphous calcium phosphate layer was formed on the surface of the β-wollastonite sample. A cell viability and proliferation assay test was performed on the rice husk ash, calcined limestone, and β-wollastonite samples by scanning electron microscope. For heavy metal element evaluation, a metal panel that included As, Cd, Pb, and Hg was selected, and both precursor and β-wollastonite fulfilled the requirement of an American Society for Testing and Materials (ASTM F1538-03) standard specification. Apart from that, a degradation test showed that the loss of mass increased incrementally as a function of soaking period. These results showed that the β-wollastonite materials produced from rice husk ash and limestone possessed good bioactivity, offering potential for biomedical applications.
Conventional delivery of anticancer drugs is less effective due to pharmacological drawbacks such as lack of aqueous solubility and poor cellular accumulation. This study reports the increased drug loading, therapeutic delivery, and cellular accumulation of silibinin (SLB), a poorly water-soluble phenolic compound using a hydrophobically-modified chitosan nanoparticle (pCNP) system. In this study, chitosan nanoparticles were hydrophobically-modified to confer a palmitoyl group as confirmed by 2,4,6-Trinitrobenzenesulfonic acid (TNBS) assay. Physicochemical features of the nanoparticles were studied using the TNBS assay, and Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) analyses. The FTIR profile and electron microscopy correlated the successful formation of pCNP and pCNP-SLB as nano-sized particles, while Dynamic Light Scattering (DLS) and Field Emission-Scanning Electron Microscopy (FESEM) results exhibited an expansion in size between pCNP and pCNP-SLB to accommodate the drug within its particle core. To evaluate the cytotoxicity of the nanoparticles, a Methylthiazolyldiphenyl-tetrazolium bromide (MTT) cytotoxicity assay was subsequently performed using the A549 lung cancer cell line. Cytotoxicity assays exhibited an enhanced efficacy of SLB when delivered by CNP and pCNP. Interestingly, controlled release delivery of SLB was achieved using the pCNP-SLB system, conferring higher cytotoxic effects and lower IC50 values in 72-h treatments compared to CNP-SLB, which was attributed to the hydrophobic modification of the CNP system.
Bacterial infections have remained significant despite our advances in the development of a plethora of disinfectants as well as antimicrobial chemotherapy. This is in part due to our incomplete understanding of the prevalence of bacterial pathogens in the environmental and clinical settings. Several lines of evidence suggest that Acanthamoeba is one of the most ubiquitous/resilient protists that also acts as a host/reservoir for pathogenic microbes. Thus targeting the hardy host, which harbour microbial pathogens, offer a potential avenue to counter infection transmission, particularly hospital/community-acquired infections. This will complement existing approach of applying disinfectants that are targeted against bacterial pathogens directly.
Blastocystis sp. is known to be the most commonly found intestinal protozoan parasite in human fecal surveys and has been incriminated to cause diarrhea and abdominal bloating. Binary fission has been widely accepted as the plausible mode of reproduction for this parasite. The present study demonstrates that subjecting the parasites in vitro to higher temperature shows the proliferation of parasite numbers in cultures. Transmission electron microscopy was used to compare the morphology of Blastocystis sp. subtype 3 isolated from a dengue patient having high fever (in vivo thermal stress) and Blastocystis sp. 3 maintained at 41 °C (in vitro thermal stress) and 37 °C (control). Fluorescence stains like acridine orange (AO) and 4',6'-diamino-2-phenylindole (DAPI) were used to demonstrate the viability and nuclear content of the parasite for both the in vitro and in vivo thermal stress groups of parasites. Blastocystis sp. at 37 °C was found to be mostly vacuolar whereas the in vitro thermal stressed isolates at 41 °C were granular with electron dense material seen to protect the granules within the central body. Parasites of the in vivo thermal stressed group showed similar ultrastructure as the in vitro ones. AO and DAPI staining provided evidence that these granules are viable which develop into progenies of Blastocystis sp. These granular forms were then observed to rupture and release progenies from the mother cells whilst the peripheral cytoplasmic walls were seen to degrade. Upon exposure to high temperature both in vitro and in vivo, Blastocystis sp. in cultures show higher number of granular forms seen to be protected by the electron dense material within the central body possibly acting as a protective mechanism. This is possibly to ensure the ability to survive for the granules to be developed as viable progenies for release into the host system.
A chitosan/polyvinyl alcohol (PVA)/zeolite composite was fabricated in this study. The composite was analyzed through field emission scanning electron microscopy, Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), thermogravimetric analysis, and weight loss test. FTIR and XRD results revealed a strong interaction among chitosan, PVA, and zeolite. Weight loss test results indicated that the composite was stable in acidic and basic media. Congo red was removed through flocculation, and the removal rate was 94% at an initial concentration of 100mg/L for a dose of 1g/L. The removal rate of methyl orange was controlled by adsorption at an initial concentration of less than 100mg/L. Flocculation occurred at high concentrations. The removal rate was also 94% at an initial concentration of 500mg/L for a dose of 5g/L. The adsorption behavior of the composite for the removal of methyl orange and Cr(VI) was described by using a pseudo-second-order kinetic model. The adsorption capacity of the composite for Cr(VI) was 450mg/g. Therefore, the synthesized composite exhibited versatility during the removal of dyes and heavy metals.
Chitin ranks next to cellulose as the most important bio-polysaccharide which can primarily be extracted from crustacean shells. However, the emergence of new areas of the application of chitin and its derivatives are on the increase and there is growing demand for new chitin sources. In this study, therefore, an attempt was made to extract chitin from the house cricket (Brachytrupes portentosus) by a chemical method. The physicochemical properties of chitin and chitosan extracted from crickets were compared with commercial chitin and chitosan extracted from shrimps, in terms of proximate analysis in particular, of their ash and moisture content. Also, infrared spectroscopy, x-ray diffraction (XRD), scanning electron microscopy and elemental analysis were conducted. The chitin and chitosan yield of the house cricket ranges over 4.3%-7.1% and 2.4%-5.8% respectively. Chitin and chitosan from crickets compares favourably with those extracted from shrimps, and were found to exhibit some similarities. The result shows that cricket and shrimp chitin and chitosan have the same degree of acetylation and degree of deacetylation of 108.1% and 80.5% respectively, following Fourier transform infrared spectroscopy. The characteristic XRD strong/sharp peaks of 9.4 and 19.4° for α-chitin are common for both cricket and shrimp chitin. The percentage ash content of chitin and chitosan extracted from B. portentosus is 1%, which is lower than that obtained from shrimp products. Therefore, cricket chitin and chitosan can be said to be of better quality and of purer form than commercially produced chitin and chitosan from shrimp. Based on the quality of the product, chitin and chitosan isolated from B. portentosus can replace commercial chitin and chitosan in terms of utilization and applications. Therefore, B. portentosus is a promising alternative source of chitin and chitosan.