Use of a modified Robbins device to directly compare the adhesion of Staphylococcus epidermidis RP62A to surfaces

Staphylococcus epidermidis is a frequent cause of infection associated with the use of biomedical devices. Flow cell studies of the interaction between bacteria and surfaces do not generally allow direct comparison of different materials using the same bacterial suspension. The use of a modified Robbins Device (MRD) to compare the adhesion to different surfaces of Staph. epidermidis RP62A grown in continuous culture was investigated. Adhesion to glass was compared with siliconized glass, plasma-conditioned glass, titanium, stainless steel and Teflon. Attachment to siliconized glass was also compared with glass under differing ionic strength, and divalent cation concentrations. Both the differences in numbers adhering and changes in adhesion (slope) through the MRD were compared. There was a trend towards higher numbers adhering to the discs at the in-flow end of the MRD than at the outflow end, probably reflecting depletion of adherent bacteria in the interacting stream. Adhesion of Staph. epidermidis RP62A to siliconized glass and Teflon was reduced when compared to glass with increasing flow rates. Adhesion to stainless steel was not affected by flow rate and titanium gave a different slope of adhesion through the MRD when compared with glass, suggesting an interaction with different sub-populations within the interacting stream. Differences between siliconized glass and glass at flow rates of 300 ml h-1 were abolished by the addition of calcium or EDTA and reduced by the addition of magnesium. Increasing ionic strength reduced the statistical significance of the differences between glass and siliconized glass. Pre-conditioning of glass with pooled human plasma reduced adhesion compared with untreated glass and again gave a different slope to glass. The MRD linked to a chemostat can be used to compare directly bacterial adhesion to potential biomaterials. Variable depletion of the interacting stream should be taken into account in the interpretation of results. Divalent cation concentration, substrate properties and flow rate were important determinants of the comparative adhesion of Staph. epidermidis RP62A to surfaces.     

玻璃粉吸附核酸及其在植物核酸提取中应用的研究

为研究玻璃粉在植物核酸提取中的应用,比较了玻璃粉颗粒大小、离液盐种类及浓度、pH等条件对玻璃粉吸附核酸的影响,得出玻璃粉吸附核酸的各种最佳条件。结果表明,普通玻璃粉吸附核酸能力强于硅胶和硅藻土,玻璃粉颗粒的直径以83 μm为佳,pH 4.0时吸附效果达到最大。提取DNA时,NaCl浓度应大于3 mol/L,而提取RNA时,异硫氰酸胍大于2 mol/L就能取得很好的效果,此外,在玻璃粉吸附RNA前,需要加入50%以上的无水乙醇才能更好地吸附。利用玻璃粉制作简易纯化柱,可用于植物组织核酸提取纯化,所提取的核酸纯度高、完整性好,可用于酶切、杂交和PCR等实验。与传统方法相比,采用玻璃粉简易离心柱提取植物核酸,效果好、环保、快速、经济。     

Host-Acceptance Requirements of Melittobia digitata (Hymenoptera: Eulophidae), a Parasitoid of Mud Dauber Wasps

Behavioral studies were conducted of Melittobia digitata Dahms on Apis mellifera (L.) pupae, pupa-shaped glass dummies, flat glass, and flat glass treated with honey bee extract to determine if females use shape or chemicals to identify a host for oviposition. Response to untreated flat glass was consistently lower than that to pupa-shaped glass. Females spent much more time on a pupa-shaped glass object than on a rectangular piece of glass. Time spent antennating on the bee pupa and on the glass pupa did not differ. However, antennation response to extract-treated flat glass and untreated flat glass was lower than that on pupa-shaped glass. The addition of host extract did not increase probing on flat glass. Wasps probed and antennated the glass dummy about as much as the bee pupa but did not respond much to the rectangular glass objects, indicating that shape plays a major role in the process of host acceptance. In these experiments, only bee pupae were accepted for oviposition and never the glass objects. In further experiments, M. digitata was found to oviposit on Parafilm domes containing agar-based diet but not on domes containing only agar. Females responded to both shape and nutritional content of the host but the surface chemical cues tested were unimportant to females considering an object for oviposition.     

Adherence of streptococci to surface-modified glass

Four types of surface-modified glass were prepared. Aminopropyl glass was prepared by alkylsilylation of glass slides with gamma-aminopropyltriethoxysilane. This glass carries primary amino groups which may be protonated at pH 7.2. Owing to the presence of both positively charged ions and hydrophobic ethoxyl groups, the glass is considered to be amphipathic. Three other types of surface-modified glass slides were prepared from aminopropyl glass by forming Schiff's bases with three aldehydes: glucose, glyoxylic acid and hexanal. The aldehyde-treated slides were subsequently reduced using sodium borohydride. Thus, the surface of the glass was rendered hydrophilic, ampholytic or hydrophobic, respectively. The adherence of two Streptococcus sanguis strains and two Streptococcus mutans strains to the surface-modified glass slides was studied. Different strains showed differences in adherence to these slides depending on their physico-chemical surface properties. For S. sanguis ATCC 10556, hydrophobic bonds seemed to be most important, while in S. mutans OMZ 176, ionic interactions made the highest contribution to adhesion. Hydrogen bonds seemed to contribute least to adherence.     

Observations on estuarine microfouling using the scanning electron microscope

Scanning electron microscopy was used to observe microbiological primary fouling of glass surfaces exposed in estuarine waters. Observations on clean glass, and glass treated with water-repellent coatings, showed that bacterial slimes adhered less strongly to the waterrepellent glass. An experiment using pure cultures of bacteria and latex particles showed that attached bacteria promoted the settlement of latex particles on the glass.     

Composition,Production and Procurement of Glass at San Vincenzo al Volturno: An Early Medieval Monastic Complex in Southern Italy

136 glasses from the ninth-century monastery of San Vincenzo and its workshops have been analysed by electron microprobe in order to situate the assemblage within the first millennium CE glass making tradition. The majority of the glass compositions can be paralleled by Roman glass from the first to third centuries, with very few samples consistent with later compositional groups. Colours for trailed decoration on vessels, for vessel bodies and for sheet glass for windows were largely produced by melting the glass tesserae from old Roman mosaics. Some weakly-coloured transparent glass was obtained by re-melting Roman window glass, while some was produced by melting and mixing of tesserae, excluding the strongly coloured cobalt blues. Our data suggest that to feed the needs of the glass workshop, the bulk of the glass was removed as tesserae and windows from a large Roman building. This is consistent with a historical account according to which the granite columns of the monastic church were spolia from a Roman temple in the region. The purported shortage of natron from Egypt does not appear to explain the dependency of San Vincenzo on old Roman glass. Rather, the absence of contemporary primary glass may reflect the downturn in long-distance trade in the later first millennium C.E., and the role of patronage in the “ritual economy” founded upon donations and gift-giving of the time.     

DNA微阵列基片的化学处理

对于合成后点样的DNA微阵列 ,基片的表面化学处理非常重要。它直接影响到样品与基片的结合效率 ,进而影响杂交结果。基片表面的各种化学修饰方法多种多样 ,物理吸附主要以赖氨酸包被为主。共价结合通常使用同源偶联分子或异源偶联分子 ,还可以在基片表面组装线状、分支状连接分子或包被琼脂糖。着重介绍了DNA微阵列的制备 ,即样品如何固定到玻璃基片上。总结了不同类型基片表面的化学修饰方法以及DNA与基片的化学结合。     

A Calorimetric Study of the Glass Transition Behaviors in Axes of Bean Seeds with Relevance to Storage Stability

Although the presence of intracellular aqueous glasses has been established in seeds, their physiological role in storage stability is still conjectural. Therefore, we examined, using differential scanning calorimetry, the thermal behavior of glass transitions in axes of bean (Phaseolus vulgaris L.) with water contents (WC) between 0 and 1 g H2O/g dry weight (g/g) and temperatures between -120 and +120[deg]C. Three types of thermal behaviors associated with the glass transition were observed. The appearance, the glass -> liquid transition temperature, and the amount of energy released during these transitions were dependent on the tissue WC. No glass transitions were observed at WC lower than 0.03 and higher than 0.45 g/g. A brief exposure to 100[deg]C altered the glass properties of tissues with WC between 0.03 and 0.08 g/g but did not affect the thermal behavior of glasses with higher WC, demonstrating that thermal history is important to the intracellular glass behavior at lower WC. Correspondence of data from bean to models predicting the effects of glass components on the glass -> liquid transition temperature suggests that the intracellular glasses are composed of a highly complex sugar matrix, in which sugar and water molecules interact together and influence the glass properties. Our data provide evidence that additional glass properties must be characterized to understand the implications of a glassy state in storage stability of seeds.     

Fungal biodeterioration of stained-glass windows

Biodeterioration of stained-glass windows by fungi was studied using historically accurate glass production methods. Glass reproductions were made according to the chemical composition determined by micro energy dispersive X-Ray Fluorescence of two historical glass windows belonging to King Ferdinand II's collection dating from the 15th and 17th centuries. Three distinct glasses compositions with different colours were selected and reproduced: i) a mixed-alkali colourless glass: ii) a purple potash-glass with manganese as chromophore, and iii) a brown potash-glass coloured by iron ions. The reproduced glass samples, with two initial surface morphologies (corroded and non-corroded), were inoculated with fungi previously isolated and identified on the original stained-glass windows as species of the genera Penicillium and Cladosporium. Physical and chemical glass surface alterations were analysed by means of optical microscopy, Raman microscopy, micro Infrared spectroscopy, and scanning electron microscopy with energy dispersive spectroscopy analysis. Results showed that fungi produced clear damage on all glass surfaces, present as spots and stains, fingerprints, biopiting, leaching and deposition of elements, and formation of biogenic crystals. Therefore, the inoculated fungi were able to biodeteriorate glasses with distinct compositions. Regarding the biodeterioration degree, there were no differences between the initial non-corroded and corroded glass surfaces.     

Enthalpy relaxation of freeze concentrated sucrose-water glass

The enthalpy relaxation of freeze concentrated sucrose-water glass was investigated using 40% sucrose, differential scanning calorimetry (DSC) with isothermal ageing for 1-6 days at various temperatures (-70, -65, -60, and -55 degrees C). The enthalpy relaxation was observed as an endothermic peak superimposed on the endothermic step-wise change due to the glass transition around -47 degrees C. The enthalpy relaxation was found to increase with ageing time and temperature. An 80% sucrose glass was also investigated at ageing temperatures of -60 and -65 degrees C, and this material exhibited a similar glass transition and enthalpy relaxation to that observed with the frozen 40% sucrose solution. The calculated activation energy of the enthalpy relaxation of the sucrose-water glass was smaller than that reported for pure sucrose. These results suggest that the freeze concentrated sucrose-water glass could have a higher molecular mobility and less stability than pure sucrose glass.     

Fabrication of Gold Nanostructures and Studies of Their Morphological and Surface Plasmonic Properties

We report fabrication of gold nanostructures on glass and indium tin oxide (ITO)-coated glass substrates using high fluence and highly energetic gold ions generated by hot, dense, and strongly non-equilibrium plasma. Nanodots and nanorods are observed in scanning electron microscopy (SEM) of nanostructures grown on glass substrate with single and double shots of gold ions which is in conformity with the transmission electron microscopy image. SEM images for single and double shots of gold ions on ITO-coated glass substrate show only nanodots. The mean diameter of nanodots obtained on both glass and ITO-coated glass is found to increase with increase in the number of gold ions shot from one to two. The gold nanostructures exhibit red shift in surface plasmon resonance with increased interaction which is in agreement with other reported work.     

Biodeteriorative processes on glass: experimental proof of the role of fungi and cyanobacteria

Biodeterioration of glass under the influence of fungi and cyanobacteria was simulated on model glasses produced according to the old recipes. Strains of fungi and cyanobacteria chosen for the investigation were isolated from biodeteriorated glass windows or similar indoor environment and are reported to be frequently involved in glass alteration. Growth of fungi and cyanobacteria resulted in the dense colonisation of the material with an expressed biofilm formation on the glass surface. The following deterioration phenomena were observed: micropitting and crack formation by all studied fungi and cyanobacteria; delineatingtraces of cells, hyphae and filaments on the glass surface; colour change of the surface due to fungal or cyanobacterial growth; biogenic minerals deposition as a consequence of the microbial metabolism on the glass surface. The pattern of glass biopitting produced in the experiment was very similar to the biopits observed on antique and medieval glasses (Krumbein et al., 1991). Crack formation pattern was strain-specific, but appeared to be independent of the chemical composition of the glass itself. The degree of deterioration was changing according to the sensitivity of the glass in question to corrosion.     

Gliding motility of Mycoplasma mobile can occur by repeated binding to N-acetylneuraminyllactose (sialyllactose) fixed on solid surfaces

Mycoplasma mobile relies on an unknown mechanism to glide across solid surfaces including glass, animal cells, and plastics. To identify the direct binding target, we examined the factors that affect the binding of Mycoplasma pneumoniae to solid surfaces and concluded that N-acetylneuraminyllactose (sialyllactose) attached to a protein can mediate glass binding on the basis of the following four lines of evidence: (i) glass binding was inhibited by N-acetylneuraminidase, (ii) glass binding was inhibited by N-acetylneuraminyllactose in a structure-dependent manner, (iii) binding occurred on glass pretreated with bovine serum albumin attached to N-acetylneuraminyllactose, and (iv) gliding speed depended on the density of N-acetylneuraminyllactose on glass.     

多聚左旋赖氨酸包被玻片的可视化检测方法

分子量15万—30万的多聚左旋赖氨酸(Poly-L-Lysine, PLL)常被用于盖玻片的包被,以促进培养细胞贴壁或切片组织黏附。然而,用市场上劣质的PLL将使组织或细胞不能牢固黏附在包被玻片上,进而导致实验失败。因而,建立实用直观的PLL质量检测方法,对于PLL包被玻片的质量控制至关重要。然而,目前世界上未见相关方法报道。因此,作者本着实用原则建立了一种实验室可视化检测盖玻片表面PLL的方法,这种方法利用化学反应将异硫氰酸荧光素(Fluorescein isothiocyanate, FITC)添加到PLL的侧链氨基(-NH2)上,再利用荧光成像检测附着在玻片表面的FITC-PLL,同时荧光照片也可以通过Fiji软件进行分析,获得定量的检测结果。利用此方法,研究发现利用不同浓度的PLL包被盖玻片后, PLL偶联的FITC荧光强度和PLL浓度成正相关。通过检测市面上的两种商品化PLL,与对照相比,两种商品化PLL附着密度极低,推断利用这两种不合格的PLL包被玻片,将导致切片样本从玻片上脱落。此方法适用于实验室自制或商品化PLL黏附盖玻片质量的可视化检测,也为附着在玻璃或塑料表面的带有...     

A method of removing gels from narrow-diameter glass tubes as part of a mini-two-dimensional electrophoresis procedure

A glass chamber that facilitates the removal of polyacrylamide gels from narrow glass tubes after the completion of isoelectric focusing is described. The tube is firmly held at one end of the glass chamber and a syringe is connected to the other end. By slowly forcing water from the syringe into the glass chamber, sufficient pressure can be generated to cause the rod gel to move slowly out of the glass tube. The rod gel is then used for the second stage of a mini-two-dimensional electrophoresis technique. Rod gels of 1.7-mm diameter have been used most extensively and can be removed from the glass tubes with almost 100% success.     

Factors Affecting the Chemical Durability of Glass Used in the Pharmaceutical Industry

Delamination, or the generation of glass flakes in vials used to contain parenteral drug products, continues to be a persistent problem in the pharmaceutical industry. To understand all of the factors that might contribute to delamination, a statistical design of experiments was implemented to describe this loss of chemical integrity for glass vials. Phase I of this study focused on the effects of thermal exposure (prior to product filling) on the surface chemistry of glass vials. Even though such temperatures are below the glass transition temperature for the glass, and parenteral compounds are injected directly into the body, data must be collected to show that the glass was not phase separating. Phase II of these studies examined the combined effects of thermal exposure, glass chemistry, and exposure to pharmaceutically relevant molecules on glass delamination. A variety of tools was used to examine the glass and the solution contained in the vial including: scanning electron microscopy and dynamic secondary ion mass spectroscopy for the glass; and visual examination, pH measurements, laser particle counting, and inductively coupled plasma–optical emission spectrometry for the analysis of the solution. The combined results of phase I and II showed depyrogenation does not play a significant role in delamination. Terminal sterilization, glass chemistry, and solution chemistry are the key factors in the generation of glass flakes. Dissolution of silica may be an effective indicator that delamination will occur with a given liquid stored in glass. Finally, delamination should not be defined by the appearance of visible glass particulates. There is a mechanical component in the delamination process whereby the flakes must break away from the interior vial surface. Delamination should be defined by the observation of flakes on the interior surface of the vial, which can be detected by several other analytical techniques.     

Patch electrode glass composition affects ion channel currents.

The influence of patch electrode glass composition on macroscopic IV relations in inside-out patches of the cGMP-activated ion channel from rod photoreceptors was examined for a soda lime glass, a Kovar sealing glass, a borosilicate glass, and several soft lead glasses. In several glasses the shape or magnitude of the currents changed as the concentration of EGTA or EDTA was increased from 200 microM to 10 mM. The changes in IV response suggest that, at low concentrations of chelator, divalent cations are released from the electrode glass and interact with the cGMP-activated channel. Possible mechanisms are discussed to explain the observations, and several comments are made concerning the choice of a glass for patching.     

Capture of genomic DNA on glass microscope slides

It is well known that DNA strands bind to silica surfaces in the presence of high concentrations of chaotropic salts. We developed simple methods to evaluate binding and recovery of DNA on flat glass microscope slides and compared their properties with commercially available silica "spin columns". Surprisingly, genomic DNA was recovered efficiently from untreated glass slides. Binding and elution times from glass slides were optimized in experiments with DNA samples of various sizes and defined buffers. Average DNA recovery from 500 ng of input genomic DNA varied from 25 to 53% for the glass slide protocol. Yields were comparable to those of spin columns. Human serum albumin and plasma components decreased DNA binding to glass several-fold in a concentration-dependent manner. These results support the concept of using flat glass slides as DNA purification surfaces in microfluidic devices for PCR sample preparation.     

Alteration of a Submarine Basaltic Glass under Environmental Conditions Conducive for Microorganisms: Growth Patterns of the Microbial Community and Mechanism of Palagonite Formation

In basaltic glass from the southern Mid-Atlantic-Ridge conducive environmental conditions for biogenic weathering resulted in excellent preserved microbial morphologies on glass surfaces. The distinct glass interface and open spaces between palagonite sheet and glass indicate a dissolution-reprecipitation mechanism of glass alteration potentially supported by microorganisms. On internal fracture surfaces, branching channels with widths at 20–30 μm containing longish structures with targeted dissolution of the glass by growing tips were observed. Alteration resulted in enrichment of Fe, Ti, P, and K in palagonite in amorphous mineral forms.     

Late Byzantine mineral soda high alumina glasses from Asia Minor: a new primary glass production group

The chemical characterisation of archaeological glass allows the discrimination between different glass groups and the identification of raw materials and technological traditions of their production. Several lines of evidence point towards the large-scale production of first millennium CE glass in a limited number of glass making factories from a mixture of Egyptian mineral soda and a locally available silica source. Fundamental changes in the manufacturing processes occurred from the eight/ninth century CE onwards, when Egyptian mineral soda was gradually replaced by soda-rich plant ash in Egypt as well as the Islamic Middle East. In order to elucidate the supply and consumption of glass during this transitional period, 31 glass samples from the assemblage found at Pergamon (Turkey) that date to the fourth to fourteenth centuries CE were analysed by electron microprobe analysis (EPMA) and by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). The statistical evaluation of the data revealed that the Byzantine glasses from Pergamon represent at least three different glass production technologies, one of which had not previously been recognised in the glass making traditions of the Mediterranean. While the chemical characteristics of the late antique and early medieval fragments confirm the current model of glass production and distribution at the time, the elemental make-up of the majority of the eighth- to fourteenth-century glasses from Pergamon indicate the existence of a late Byzantine glass type that is characterised by high alumina levels. Judging from the trace element patterns and elevated boron and lithium concentrations, these glasses were produced with a mineral soda different to the Egyptian natron from the Wadi Natrun, suggesting a possible regional Byzantine primary glass production in Asia Minor.