Flow cytometric evaluation of sperm apoptosis in semen of silver foxes in the breeding period

The objective of the study was to evaluate cytometrically the percentage of apoptotic and necrotic spermatozoa in fresh semen of silver foxes in the breeding season. In males F3 and F4 with high percentages of early apoptotic (A+Pi−), late apoptotic (A+Pi+) and necrotic (A−Pi+) spermatozoa as well as 56–65% of living spermatozoa (A−Pi−) with progressive motility, the semen was characterised by reduced fertility. In males F1 and F2 with spermatozoa showing the motility and viability of 89–90% and high percentages of living cells that do not bind Annexin V and propidium iodide, the semen was assessed as valuable and useful for artificial insemination. Amongst 16 females of group I and II inseminated with semen from F1 and F2 males, 15 (93.75%) had multi-cub litters – on average 6.1 and 4.8, respectively. In contrast, amongst 16 females of group III and IV inseminated with semen from F3 and F4 males, only 10 (62.5%) had litters with few cubs (on average 2.6 in group III and 2.1 in group IV). Our findings explicitly indicate that semen of farm male foxes should be evaluated before the breeding season, as one of the causes of reproduction failures is likely to be a high percentage of apoptotic and necrotic spermatozoa. Thanks to flow cytometry, fresh ejaculates can be speedily evaluated and their usefulness for artificial insemination determined.     

A silvering index for the Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

To establish a simple and reliable index for determining silvering stages of the Japanese eel, Anguilla japonica, we observed the colorations of various body parts and biological characteristics of the eels collected in a coastal area of Japan (Mikawa Bay). The four silvering stages are characterized by the colorations of pectoral fins and ventral skin as follows: (1) Y1, yellow eel without a metallic hue at the base of pectoral fins, (2) Y2, late yellow eel with a metallic hue at the base of the pectoral fins but without melanization at the tip of pectoral fins, (3) S1, silver eel with complete melanization at the tip of pectoral fins but without full pigmented belly in black or dark brown, and (4) S2, late silver eel with black or dark brown belly. The body size, eye diameter and sexual maturity of each stage increased in the order of Y1, Y2, S1 and S2 stages, whereas the digestive tract degenerated in the same order, suggesting a sequential development of these ontogenetic stages identified in the study. The Y1, Y2 and S1 stages could be also distinguished by canonical discriminant function analysis using three internal (gonad-somatic index, GSI; hepato-somatic index, HIS; and gut index) and two morphometric (condition factor and eye index) parameters, supporting the significance of these stages. This method of staging for the silvering process of the Japanese eel appeared to be applicable to all specimens of this species, since this index used only simple external characteristics that would be easy to observe during field surveys.     

Preparation and Optical Characterization of Core–Shell Bimetal Nanoparticles

Chemical approaches allow for the synthesis of highly defined metal heteronanostructures, such as core–shell nanospheres. Because the material in the metal nanoparticles determines the plasmon resonance-induced absorption band, control of particle composition results in control of the position of the absorption band. Metal deposition on gold or silver nanoparticles yielded core–shell particles with modified optical properties. UV–vis spectroscopy on solution-grown, as well as surface-grown, particles was conducted and provided ensemble measurements in solution. Increasing the layers of a second metal leads to a shift in the absorption band. A shell diameter comparable to the original particle diameter leads to a predominant influence by the shell material. Extent of shell growth could be controlled by reaction time or the concentration of metal salt or reducing agent. Besides optical characterization, the utilization of atomic force microscopy, scanning electron microscopy, and transmission electron microscopy yielded important information about the ultrastructure of nanoparticle complexes. Surface-grown core–shell particles were superior in terms of achievable shell thickness, because of difficulties encountered with solution-grown particles due to salt-induced aggregation.     

A versatile tool in controlling aggregation and Ag nanoparticles assisted in vitro folding of thermally denatured zDHFR

BackgroundProteins have tendency to form inactive aggregates at higher temperatures due to thermal instability. Maintenance of thermal stability is essential to gain the protein in sufficient quantity and biologically active form during their commercial production.MethodsBL21-DE3 Rosetta E. coli cells which contains plasmid pET43.1a vector was used for producing zDHFR protein commercially. The purification of N-terminal Histidine tagged zDHFR was performed by Immobilized Metal Ion chromatography (IMAC). Investigations were performed in existence and non existence of Silver nanoparticles (AgNPs). The inactivation kinetics of zDHFR in existence and non existence of AgNPs were monitored over a range of 40–80 °C as monitored by UV–Visible absorption spectroscopy.ResultsThe protein completely lost its activity at 55 °C. Kinetics of inactivated zDHFR follows first order model in presence and absence of AgNPs. Decrease in rate constant (k) values at respective temperatures depicts that AgNPs contribute in the thermostability of the protein. AgNPs also assists in regaining the activity of zDHFR protein.ConclusionsAgNPs helps in maintaining thermostability and reducing the aggregation propensity of zDHFR protein.General significanceResult explains that AgNPs are recommended as a valuable system in enhancing the industrial production of biologically active zDHFR protein which is an important component in folate cycle and essential for survival of cells and prevents the protein from being aggregated.     

Antibiofilm and antimicrobial activities of green synthesized silver nanoparticles using marine red algae Gelidium corneum

In our study, green synthesis of silver nanoparticles was carried out using a red algae Gelidium corneum extract as reducing agent. The obtained silver nanoparticles were characterized by UV–vis, TEM, XRD, FTIR and ICP-MS measurements. FTIR measurements indicated the possible functional groups responsible for the stabilization and reduction of nanoparticles, while XRD analysis results explained the crystalline structure of the particles with centric cubic geometry. TEM micrographs showed that the size of the nanoparticles was between 20–50 nm. According to the broth microdilution test results, AgNPs showed a high antimicrobial activity with very low MIC values (0.51 μg/ml for Candida albicans yeast and 0.26 μg/ml for Escherichia coli bacteria). The different ultrastructural effects of silver nanoparticles on yeast and bacterial cells were observed by TEM. Antibiofilm efficacy studies were also examined in two stages as prebiofilm and postbiofilm effect. In prebiofilm effect studies, AgNPs (0.51 μg/ ml) exhibited 81% reducing effect on biofilm formation. The highest reduction rate in postbiofilm studies was 73.5% and this was achieved with 2.04 μg/ml AgNPs. Our data support that the silver nanoparticles obtained by this environmentally friendly process have potential to be used for industrial and therapeutic purposes.     

Preparation of silver nanoparticles and riboflavin embedded electrospun polymer nanofibrous scaffolds for in vivo wound dressing application

Biocompatible silver-based nanofibrous frameworks have attracted intensive attention in wound dressing materials ascribed to their greater stability, minimal toxicity, excellent antibacterial activity, and extended therapeutic efficiency. The present investigation delineates a simple approach to synthesize silver nanoparticles (Ag NPs), and riboflavin (RF) decorated polyvinyl alcohol/β-Cyclodextrin (PVA/β-CD) electrospun nanofibrous scaffolds envisioning their application in wound dressings. PVA/β-CD polymer matrix regulates the stabilization of Ag NPs and RF. Also, it promotes the wound healing process and skin regeneration. The morphology, thermal properties, and their structure were also evaluated. Likewise, mechanical properties, biodegradation and drug release profile of the nanofibrous scaffolds were evaluated. In addition Antibacterial studies of the resultant nanofibrous scaffolds showed a strong inhibitory effect against Staphylococcus aureus and Escherichia coli at a considerable level. Moreover, Ag NPs-RF/PVA/β-CD nanofibrous scaffold were studied for its in vitro cytotoxicity using human embryonic kidney cells (HEK-293), and the results suggested that Ag NPs and RF present in the nanofibrous scaffolds exhibited its cytotoxicity. Besides, wound healing efficiency of the Ag NPs-RF decorated nanofibrous scaffolds was assessed using full thickness excision wounds in rat models displayed as an excellent biomaterial for wound dressings.     

Gold and silver complexes with the ferrocenyl-pyrazolate ligand FcCH2pz

We have synthesized a series of gold(I), gold(III) or silver(I) derivatives with ferrocenylmethylpyrazole (pzCH₂Fc) as ligand. X-ray structure determination for [Au(C₆F₅)(pzCH₂Fc)] reveals a linear geometry for the gold centre and association of the molecules in pairs through intermolecular gold(I)-gold(I) interactions of 3.1204(6) Å. These molecules are further assembled into chains and the chains into a 3D structure through several hydrogen bonds. [Ag(OTf)(pzCH₂Fc)] (OTf = triflate) is a dimer with a distorted tetrahedral geometry at the silver(I) centre and the triflate acting as bridging ligand.     

Effect of gemini surfactant (16-6-16) on the synthesis of silver nanoparticles: A facile approach for antibacterial application

In this report, we describe the effect of Gemini surfactants1, 6-Bis (N, N-hexadecyldimethylammonium) adipate (16-6-16) on synthesis, stability and antibacterial activity of silver nanoparticles (AgNPs). The stabilizing effect of Gemini surfactant and aggregation behavior of AgNPs was evaluated by plasmonic property and morphology of the AgNPs were characterized by UV–vis spectroscopy, Dynamic Light Scattering (DLS), X-ray diffraction (XRD), High resolution transmission electron microscopy (HRTEM) and Energy dispersive X-ray analysis (EDX) techniques. Interestingly, the formation of quite mono-dispersed spherical particles was found. Apart from the stabilizing role, the Gemini surfactant has promoted the agglomeration of individual AgNPs in small assemblies whose Plasmon band features differed from those of the individual nanoparticles. The antibacterial activity of the synthesized AgNPs on Gram-negative and Gram-positive bacterium viz., E. coli and S. aureus was carried out by plate count, growth kinetics and cell viability assay. Furthermore, the mechanism of antibacterial activity of AgNPs was tested by Zeta potential and DLS analysis, to conclude that surface charge of AgNPs disrupts the cells causing cell death.     

Production and Characterization of Protein Encapsulated Silver Nanoparticles by Marine Isolate Streptomyces parvulus SSNP11

Production of protein encapsulated silver nanoparticles (AgNPs) assisted by marine actinomycetes strain has been investigated. The selective isolate was identified as Streptomyces parvulus SSNP11 based on chemotaxonomic and 16S rRNA analysis. Maximum AgNPs production was observed within 24 h incubation time. The produced AgNPs are spherical in shape with monodispersive and crystalline in nature. The particle size distribution ranges from 1.66 to 11.68 nm with a mean size of 2.1 nm. The biosynthesized AgNPs revealed stretching vibrations of primary and secondary amines along with C–H and C–N, suggesting that metabolically produced proteins are involved in size regulation of reduced AgNPs. These particles possess an average negative zeta potential value of 81.5 mV with an electrophoretic mobility of 0.000628 cm²/Vs. The biosynthesized nanoparticles revealed antimicrobial property against gram negative as well as gram positive bacterial strains.     

Influence of Silver Colloid Presence on Stilbazolium Merocyanine Emission

Absorption, fluorescence emission, and fluorescence excitation spectra of stilbazolium merocyanine (1-(n-butyl)-4[(3,5-dimethoxy-4-oxocyclohexa-2,5-dienylidene)ethylidene]-1,4-dihydropyridyne) dye in water solution without and with colloidal silver addition were measured. In the presence of the colloid, besides the absorption band assigned to the protonated species of the dye (at 391 nm), an absorption band related to the free-base species appears at 490 nm. From the absorption and emission spectra, measured at various dye concentrations, follows that the aggregates are not effectively formed. Therefore, the long-wavelength absorption and fluorescence bands have to be related to some dye forms created by the solvatochromic effects. The fluorescence bands of the protonated and the free-base species are located at 559 nm and at about 630 nm, respectively. The shape of the long-wavelength band suggests the occurrence of more than one free-base form of the dye. At some dye and colloid concentrations, the global emission of the sample is enhanced as a result of silver addition. The increase in the emission yield of the dye could be partially due to not only the change in the concentrations of dye forms exhibiting various emission yields but is also due to the resonance surface plasmon effect. Because of the superposition of several effects, before the practical application of merocyanine as an indicator of metal presence in biological samples, its spectral properties in the system investigated should be established.