Immobilization of Arg-Gly-Asp peptides on silk fibroin via Gly-Ala-Gly-Ala-Gly-Ser sequences.

A simple method by which the functional peptide of Gly-Arg-Gly-Asp-Ser (GRGDS) is immobilized on the surface of silk fibroin (SF) films via Gly-Ala-Gly-Ala-Gly-Ser (GAGAGS) sequences is proposed. GAGAGS, a repeating amino acid sequence in the crystal region of Bombyx mori SF, performs a key role in interacting with and immobilizing SF molecules. Immobilization by this proposed method involves no chemical reaction, thereby preserving the original properties of the SF molecule. The density of GRGDS peptides existing on SF film was found to be higher in the GAGAGS-bound type than in the non-GAGAGS-bound type. Furthermore, results showed that the amount of immobilized (GAGAGS)GRGDS peptide increased as the ß-sheet crystallization was promoted in the SF film. Fibroblasts, which adhered to the surface of the SF film, showed more extensibility because of the (GAGAGS)GRGDS immobilization, which suggests that the cell adhesion activity of RGD is functioning effectively.

Combining thermal hydrolysis and methylation-gas chromatography/mass spectrometry with X-ray photoelectron spectroscopy to characterise complex organic assemblages in geological material.

What follows is a method applicable generically to the analysis of low levels of organic matter that is embedded in either loose fine-grained or solid geological material. Initially, the range of organic compounds that could be detected in a geological sample using conventional pyrolysis chromatography/mass spectrometry was compared to the range that was detected using thermally assisted hydrolysis and methylation-gas chromatography/mass spectrometry (THM-GC/MS). This method was used to validate the synthetic components fitted to X-ray photoelectron spectroscopy (XPS) carbon spectra of the sample. Reciprocally, XPS analysis was able to identify the constituent carbon-carbon, carbon-oxygen and carbon-nitrogen bonds of the functional groups in the compounds identified by THM-GC/MS. The two independently derived outputs from the THM-GC/MS and the XPS techniques mutually validated the identification of organic compounds in our geological samples. We describe in detail the improvements to: •The preparation of geological samples for analysis by XPS.•Measurements of organic material in geological samples using GC/MS.•The use of THM-GC/MS and XPS data used together to characterise low levels of organic material in geological samples.

The organic stratigraphy of Ontong Java Plateau Tuff correlated with the depth-related presence and absence of putative microbial alteration structures.

Structures in geological samples are often interpreted as fossilised life; however, such interpretations are equivocal, as abiotic processes can be invoked to explain their presence. Thus, additional lines of chemical evidence are invaluable in confirming or refuting such morphological evidence. Glass shards in tuff from the Ontong Java Plateau (OJP) contain microtubular structures that are in close proximity to functionalised nitrogen substituted aromatic compounds that may be indicative of the chemical remnants of biological activity. The organic composition of the OJP tuff containing microtubular alteration structures was compared with tuff without such features. In addition, organic matter associated with horizons with compacted remnants of woody material buried in the OJP tuff and overlying pelagic calcareous foraminifer sediment were also characterised, to ascertain the provenance of the organic matter found in the OJP tuff. As a further control, the organic material in submarine and terrestrial basalts from other locations were also characterised providing further evidence to support the view that the organic matter in the OJP tuff is authigenic. Carbon-nitrogen chemistry was detected across all OJP tuff samples irrespective of the presence or absence of microtubular features, but was not detected in either the wood material, the overlying pelagic sediments or in the basalts from other locations. The results indicate no direct link between the OJP nitrogenous organic compounds and the presence or absence of microtubular features.

Removing Beam Current Artifacts in Helium Ion Microscopy: A Comparison of Image Processing Techniques.

The development of the helium ion microscope (HIM) enables the imaging of both hard, inorganic materials and soft, organic or biological materials. Advantages include outstanding topographical contrast, superior resolution down to <0.5 nm at high magnification, high depth of field, and no need for conductive coatings. The instrument relies on helium atom adsorption and ionization at a cryogenically cooled tip that is atomically sharp. Under ideal conditions this arrangement provides a beam of ions that is stable for days to weeks, with beam currents in the order of picoamperes. Over time, however, this stability is lost as gaseous contamination builds up in the source region, leading to adsorbed atoms of species other than helium, which ultimately results in beam current fluctuations. This manifests itself as horizontal stripe artifacts in HIM images. We investigate post-processing methods to remove these artifacts from HIM images, such as median filtering, Gaussian blurring, fast Fourier transforms, and principal component analysis. We arrive at a simple method for completely removing beam current fluctuation effects from HIM images while maintaining the full integrity of the information within the image.

Pulsatile drug release from electrospun poly(ethylene oxide)-sodium alginate blend nanofibres.

Novel and highly tuneable pulsatile drug delivery systems have been prepared through the electrospinning of a blend of poly(ethylene oxide) (PEO), sodium alginate (SA), and sodium ibuprofen (SI). The resultant fibres contain crystallites of SI embedded in a PEO-SA matrix, and rather than being obtained as flat mats on the collector plate form novel three dimensional structures extending upwards the needle. Fibres were prepared with a range of loadings of SI and SA. It was found that at pH 6.8 (reminiscent of the intestinal tract) the fibres dissolve very rapidly, freeing all the embedded drug within ca. 20 minutes. However, at pH 3 (representative of the stomach pH in the fed state or in older patients) an unusual two stage release mechanism is seen. This comprises a rapid burst release, followed by a period where no further drug is released for ca. 120-150 minutes, and then a final stage of release freeing the remainder of the drug into solution. The amount of release in the initial stage, and the length of time between the first and final drug release stages, can be controlled by adjusting the SI and SA contents of the fibres respectively. This results in highly tunable pulsatile release materials.

Acrylic-acid-functionalized PolyHIPE scaffolds for use in 3D cell culture.

This study describes the development of a functional porous polymer for use as a scaffold to support 3D hepatocyte culture. A high internal phase emulsion (HIPE) is prepared containing the monomers styrene (STY), divinylbenzene (DVB), and 2-ethylhexyl acrylate (EHA) in the external oil phase and the monomer acrylic acid (Aa) in the internal aqueous phase. Upon thermal polymerization with azobisisobutyronitrile (AIBN), the resulting porous polymer (polyHIPE) is found to have an open-cell morphology and a porosity of 89%, both suitable characteristics for 3D cell scaffold applications. X-ray photo-electron spectroscopy reveals that the polyHIPE surface contained 7.5% carboxylic acid functionality, providing a useful substrate for subsequent surface modifications and bio-conjugations. Initial bio-compatibility assessments with human hepatocytes show that the acid functionality does not have any detrimental effect on cell adhesion. It is therefore believed that this material can be a useful precursor scaffold towards 3D substrates that offer tailored surface functionality for enhanced cell adhesion.

Galactose-functionalized polyHIPE scaffolds for use in routine three dimensional culture of mammalian hepatocytes.

Three-dimensional (3D) cell culture is regarded as a more physiologically relevant method of growing cells in the laboratory compared to traditional monolayer cultures. Recently, the application of polystyrene-based scaffolds produced using polyHIPE technology (porous polymers derived from high internal phase emulsions) for routine 3D cell culture applications has generated very promising results in terms of improved replication of native cellular function in the laboratory. These materials, which are now available as commercial scaffolds, are superior to many other 3D cell substrates due to their high porosity, controllable morphology, and suitable mechanical strength. However, until now there have been no reports describing the surface-modification of these materials for enhanced cell adhesion and function. This study, therefore, describes the surface functionalization of these materials with galactose, a carbohydrate known to specifically bind to hepatocytes via the asialoglycoprotein receptor (ASGPR), to further improve hepatocyte adhesion and function when growing on the scaffold. We first modify a typical polystyrene-based polyHIPE to produce a cell culture scaffold carrying pendent activated-ester functionality. This was achieved via the incorporation of pentafluorophenyl acrylate (PFPA) into the initial styrene (STY) emulsion, which upon polymerization formed a polyHIPE with a porosity of 92% and an average void diameter of 33 µm. Histological analysis showed that this polyHIPE was a suitable 3D scaffold for hepatocyte cell culture. Galactose-functionalized scaffolds were then prepared by attaching 2'-aminoethyl-ß-D-galactopyranoside to this PFPA functionalized polyHIPE via displacement of the labile pentafluorophenyl group, to yield scaffolds with approximately ca. 7-9% surface carbohydrate. Experiments with primary rat hepatocytes showed that cellular albumin synthesis was greatly enhanced during the initial adhesion/settlement period of cells on the galactose-functionalized material, suggesting that the surface carbohydrates are accessible and selective to cells entering the scaffold. This porous polymer scaffold could, therefore, have important application as a 3D scaffold that offers enhanced hepatocyte adhesion and functionality.

[Diffuse large B-cell lymphoma with intravascular involvement of neoplastic CD5-positive cells].

A 77-year-old man was admitted because of fever. A small number of large CD20-positive neoplastic cells were seen in the bone marrow specimen. Clinical symptoms improved with oral prednisolone. After 11 months, abdominal CT scan revealed a liver mass. The biopsy specimen from the liver mass showed diffuse infiltration of large CD20-positive neoplastic cells. The patient was diagnosed as having diffuse large B-cell lymphoma. It was of particular interest that only neoplastic B cells within small blood vessels in the liver mass were positive for CD5. The patient died of lymphoma three months after diagnosis.

Nanomechanical measurements of hair as an example of micro-fibre analysis using atomic force microscopy nanoindentation.

The characterisation of nanoscale surface properties of textile and hair fibres is key to developing new effective laundry and hair care products. Here, we develop nanomechanical methods to characterise fibres using an atomic force microscope (AFM) to give their nanoscale modulus. Good mounting methods for the fibre that are chemically inert, clean and give strong mechanical coupling to a substrate are important and here we detail two methods to do this. We show, for elastic nanoindentation measurements, the situation when the tip radius significantly affects the result via a function of the ratio of the radii of the tip and fibre and indicate the importance of using an AFM for such work. A valid method to measure the nanoscale modulus of fibres using AFM is thus detailed and exampled on hair to show that bleaching changes the nanoscale reduced modulus at the outer surface.

The Christiansen effect of brightly colored colloidal dispersion with an amphiphilic polymer.

A novel coloration phenomenon in a colloidal dispersion with an amphiphilic polymer was found. The dispersion consists of tetrahydrofuran (THF), an aqueous solution of sodium thiosulfate (Na(2)S(2)O(3).5H(2)O), and hydroxypropylcellulose (HPC). The dispersion was emulsified by HPC as an amphiphilic polymer, so that the aqueous phase was confined in droplets in the THF matrix. It typically appeared bluish violet at room temperature and turned into blue with increasing temperature. In this system, the refractive indices of the inside and outside of the droplet coincided at a certain wavelength at which the light passes through without scattering, which is called the Christiansen effect. The color observed was successfully simulated by Mie's scattering theory in combination with the Christiansen effect.

[Spontaneous remission of acromegaly after meningitis: a case report].

There have been several reports describing the cases of acromegaly, which show reduction in size of tumor in due to pituitary apoplexy or lymphocytic hypophysitis. We have encountered a patient of acromegaly, who developed panhypopituitarism after suffering from meningitis and showed the reduction of tumor in size. The results of imaging examination suggested the presence of pituitary apoplexy and lymphocytic hypophysitis. The patient was a 27-year-old woman, who visited a local physician with complains of headache and fever. After performing lumbar puncture, she was diagnosed as viral meningitis, and conservative therapy was initiated. The results of biochemical test of blood revealed hyponatremia. Because facial appearance of the patient was similar to that of acromegaly, endocrine dysfunction was suspected. The result of pituitary hormone tests showed high levels of growth hormone (GH) and somatomedin C (IGF-1) and low levels of the other hormones. At the same time, sign of diabetes insipidus was noted, and the patient was referred to our hospital. In the examination at the admission, GH and IGF-1 showed the trends to decrease, and the reduction in size of tumor was also detected. From the results of imaging examination, pituitary apoplexy and lymphocytic hypophysitis were suspected. Operation was performed, and pathological examination revealed inflammation of pituitary adenoma.