Metallic gold (Au°) is a likely biotransformation product of monovalent gold, Au(I) whenever it is dissociated from in vivo ligands, Au° being formed either by bioreduction or by spontaneous dismutation (with co-production of trivalent gold). This review discusses the preparation and some biologically relevant properties of colloidal metallic gold (CMG) in its nano-particulate form. Tyndall’s purple, a well characterised preparation of CMG, shows potent anti-arthritic activity in rats, approximately 103 times that of sodium aurothiomalate (Myocrysin). Even more remarkable is its broader spectrum of action in rats compared to this classic DMARD.

BACKGROUND:White matter fiber tracts, especially those interconnecting the frontal and temporal lobes, are likely implicated in pathophysiology of schizophrenia. Very few studies, however, have focused on the fornix, a compact bundle of white matter fibers, projecting from the hippocampus to the septum, anterior nucleus of the thalamus and the mamillary bodies. Diffusion Tensor Imaging (DTI), and a new post-processing method, fiber tractography, provides a unique opportunity to visualize and to quantify entire trajectories of fiber bundles, such as the fornix, in vivo. We applied these techniques to quantify fornix diffusion anisotropy in schizophrenia. METHODS:DTI images were used to evaluate the left and the right fornix in 36 male patients diagnosed with chronic schizophrenia and 35 male healthy individuals, group matched on age, parental socioeconomic status, and handedness. Regions of interest were drawn manually, blind to group membership, to guide tractography, and fractional anisotropy (FA), a measure of fiber integrity, was calculated and averaged over the entire tract for each subject. The Doors and People test (DPT) was used to evaluate visual and verbal memory, combined recall and combined recognition. RESULTS:Analysis of variance was performed and findings demonstrated a difference between patients with schizophrenia and controls for fornix FA (p=0.006). Protected post-hoc independent sample t-tests demonstrated a bilateral FA decrease in schizophrenia, compared with control subjects (left side: p=0.048; right side p=0.006). Higher fornix FA was statistically significantly correlated with DPT and measures of combined visual memory (r=0.554, p=0.026), combined verbal memory (r=0.647, p=0.007), combined recall (r=0.516, p=0.041), and combined recognition (r=0.710, p=0.002) for the control group. No such statistically significant correlations were found in the patient group. CONCLUSIONS:Our findings show the utility of applying DTI and tractography to study white matter fiber tracts in vivo in schizophrenia. Specifically, we observed a bilateral disruption in fornix integrity in schizophrenia, thus broadening our understanding of the pathophysiology of this disease.

The aurocyanide anion, Au(CN) 2 − , is a human metabolite of several anti-rheumatic gold complexes containing monovalent gold (I) bound to a sulphur ligand. This article reviews some of the chemical and pharmacological properties of this intriguing metabolite, and reports its anti-arthritic and anti-inflammatory activity in rats. Au(CN) 2 − is generated from the therapeutic gold complexes by small amounts of hydrogen cyanide, HCN, produced from thiocyanate, SCN−, by myeloperoxidase (MPO) an enzyme in neutrophils which normally produces hypochlorite, OCl−. Thus, Au(CN) 2 − is formed at sites of inflammation where activated neutrophils are present. This includes atherosclerotic lesions as well as inflamed joints. MPO also oxidises Au(CN) 2 − to Au(III) complexes such as Au(CN) 4 − .

Most current high throughput purification procedures for the green fluorescent protein (GFP) suffer from poor yields and low purity. An improved purification procedure that delivers highly pure protein (>95% homogeneity) in high yields (>70% of the initial fluorescent protein content) has been developed. The purification procedure requires only two steps: the cell lysate is heated to 60°C for 4min in ammonium sulfate and triethylamine, followed by hydrophobic interaction chromatography using isopropanol during the elution phase. The resulting pure product exhibits the same fluorescence profile as the crude sample. This procedure has been demonstrated on three commercial variants of GFP from Aequorea victoria, enhanced green, enhanced yellow, and enhanced cyan fluorescent protein (Becton–Dickinson). The yield and purity of material are superior to other recently described methods.

The structure of human fibroblasts has been characterised in vitro by atomic force microscopy (AFM) operated in the imaging or in the force vs. distance (F-d) modes. The choice of growth substrate is important to ensure good adhesion. The substrate also affects the imaging conditions for in vitro analysis of live cells; activated tissue culture dishes are shown to promote conditions that routinely result in good quality images. A qualitative model suggests that the activated substrate may act as a preferential scavenger of cellular debris, therefore promoting low adhesion between tip and membrane and preventing the tip from biofouling. Alternatively, the activated substrate may promote a more rigid cell structure, thus resulting in improved imaging. Good imaging conditions provide nondestructive in vitro information about cytoskeletal structure and dynamics; thus, treatment with cytochalasin can be monitored in real time for durations of several hours. (C) 1999 Elsevier Science B.V. All rights reserved.

The structure of human fibroblasts have been characterised in vitro by atomic force microscopy (AFM) operated in the imaging or in the force versus distance (F-d) modes. The choice of cell substrate is important to ensure good adhesion. Of greater significance in the context of AFM analysis, is the observation that the substrate affects the imaging conditions for in vitro analysis of live cells. For instance, very rarely will glass coverslips lead to acceptable outcomes (i.e., resolved cytoskeletal structure). Activated tissue culture dishes, on the other hand, promote conditions that routinely result in good quality images. Those conditions are then unaffected by adoption of relatively high force loadings (more than 10 nN), large fields of view (100 x 100 microm2) and high scan speeds (up to ca. 200 microm/sec), all of which exceed values recommended in the literature. Plasma membranes are fragile in the context of AFM analysis (F-d analysis gives an equivalent Young's Modulus of ca. 5 kPa). However, the present work suggests that fragility per se need not be a problem, rather it is the adhesive interactions with the tip, which under some circumstances may exceed 20 nN, that are the source of poor imaging conditions. The present results, being supported by a qualitative model, suggest that the activated substrate acts as a preferential scavenger of cellular debris thus preventing the tip from biofouling, and will therefore promote low adhesion between tip and membrane. Good imaging conditions provide non-destructive in vitro information about cytoskeletal structure and dynamics, as shown in two examples concerned with cytochalasin treatment and with the MTT assay.

ABSTRACT: The olfactory epithelium is a unique system, in which new neurons are continually generated throughout adult life. Olfactory neurons are derived from stem cells that lie adjacent to the basal lamina of the olfactory epithelium; these stem cells divide several times and their progeny differentiate into mature sensory neurons. In our laboratory immortalized cell lines have been derived from these dividing cells. The morphology of these cell lines and their expression of neuronal markers varies with culture conditions. When grown in low serum medium one of these cells lines, OLF 442, differentiates by extending long neurites and increasing its expression of neurofilament and B50/GAP43 proteins at the same time reducing expression of glial fibrillary acidic protein (GFAP). Identification of differentially expressed mRNA in cell lines has previously relied on both screening for known markers, and the use of subtractive techniques for identification of unique mRNA species. The differential display technique allows simultaneous detection of differentially expressed mRNA at different time periods and growth conditions. A modified Liang and Pardee differential display technique was used to screen OLF 442 over a number of time intervals in serum‐depleted media, and compared with OLF 442 grown in complete media. The differentially displayed fragments were cloned and sequenced, leading to the identification of a number of sequences, both known and unknown. The known sequences include SPARC (encoding a Ca 2+ binding secreted Protein which is Acidic and Rich in Cysteine), which is reported to function as a modulator of the cell matrix, and RHAMM, the receptor for hyaluronan‐mediated motility. Both the known and the unknown sequences are being studied further to provide insight into the differentiation of olfactory neurons.

Being genetically homogeneous, clonal cell lines are potentially important for investigating many aspects of cellular differentiation. We describe here the creation of clonal cell lines by immortalization of neuronal precursor cells from the adult mouse olfactory epithelium. Unlike neurons elsewhere in the vertebrate nervous system, the olfactory sensory neuron can be replaced throughout the lifespan of the animal, However, little is known about the molecular aspects of olfactory neurogenesis. Continuous cell lines were generated by retroviral transduction of the n-myc proto-oncogene into the mitotically active basal cells of the olfactory epithelium which give rise to the sensory neuron, Twenty-one clonal cell lines were produced which could be divided into three distinct morphological classes: one with flat, epithelial-like cells only; another with round, flat, and bipolar cells; and a third with large flat and large bipolar cells, These morphological classes had different patterns of intermediate filament expression, as shown by immunocytochemistry and immunoblot analysis, All cells in all cell lines expressed the intermediate filament protein vimentin, Most bipolar cells, but not other cell types, expressed neurofilament protein and in one morphological class the bipolar cells co-expressed neurofilament and glial fibrillary acidic protein, Several cell lines expressed mRNA for OMP, a marker of mature olfactory sensory neurons, and G(OLF), a guanine nucleotide binding protein involved in olfactory sensory transduction, It is concluded that these cell lines were immortalized from sensory neuron precursors late in the lineage pathway, Other cell lines appear to have been immortalized at earlier stages in the lineage pathway, These cell lines therefore provide useful tools for the investigation of neuronal differentiation and sensory transduction in the olfactory epithelium. (C) 1996 Wiley-Liss, Inc.

Human parainfluenza viruses (HPIV) 1–4 had been analyzed as being one of the most frequent causes of hospitalizations for young children with respiratory tract illnesses.This retrospective study was performed from children virologically confirmed as HPIV infection through throat swab or nasopharyngeal aspirates at a tertiary care university hospital, between January 2012 and December 2014. HPIV4 was not checked and analyzed, due to not include in the commercial kit. The demographic, epidemiological, clinical presentations, diagnosis, treatment, outcomes, and laboratory data were analyzed.Totally 398 cases were enrolled, including 39 (9.8%) of HPIV1, 67 (16.8%) of HPIV2, and 292 (73.4%) of HPIV3. The mean age of HPIV-infected children was 2.9 year-old, and 50.5% were among one to three year-old. A total of 56.8% HPIV3-infected children were among one to three years old, however, no HPIV2-infected children was younger than one year-old. The HPIV1-infected patients were more common to develop wheezing and diagnose as acute bronchiolitis. HPIV2-infected children were more likely to have hoarseness (23.9%), and were associated with croup (25.4%). HPIV3 was isolated from two fatal cases, with neurological underlying diseases.The impact caused by HPIVs infections is significant in hospitalized children. In the current study, our results contribute to the epidemiologic, clinical and laboratory information of HPIV infection in children in the important areas of respiratory tract infection that could support the development of optimization management.

Neurogenesis in the adult olfactory epithelium is highly regulated in vivo. Little is known of the molecular signals which control this process, although contact with the olfactory bulb or with astrocytes has been implicated. Explants of mouse olfactory epithelium were grown in the presence or absence of several peptide growth factors. Basic fibroblast growth factor (FGF2) stimulated differentiation of sensory neurons in adult and embryonic olfactory epithelium. Other growth factors tested were ineffective. FGF2-stimulated neurons were born in vitro and expressed neurofilament, neural cell adhesion molecule, and β-tubulin. The cells also expressed olfactory marker protein, a marker for mature olfactory sensory neurons in vivo. These bipolar neurons did not express glial fibrillary acidic protein or low-affinity nerve growth factor receptor. These results indicate that neither astrocytes nor olfactory bulb are necessary for differentiation of olfactory sensory neurons in vitro. © 1996 Wiley-Liss, Inc.

This report describes neurogenesis in the adult human olfactory epithelium in vitro. Olfactory epithelium was collected at autopsy and by biopsy, and grown in serum-free medium. Basic fibroblast growth factor induced the differentiation of bipolar cells which were immunopositive for several neuronal proteins but not glial proteins. [3H]thymidine autoradiography confirmed that these neurones were born in vitro. The results demonstrate that the adult human olfactory epithelium retains the capacity for neurogenesis and neuronal differentiation, at least until the age of 72 years. It is now possible to examine neurones and neurogenesis in biopsies from patients with disorders that may involve a neurodevelopmental or neurodegenerative aetiology such as schizophrenia, bipolar disorder and Alzheimer's disease.

SummaryTobacco mosaic virus (TMV) has been deposited on freshly cleaved mica substrates. The topography was investigated by contact, non‐contact and lateral‐force microscopy under ambient conditions in air. The results were in accord with known dimensions of TMV (i.e. 18 nm in diameter and 300 nm in length). However, convolution of tip shape with TMV morphology resulted in an apparent width of 80–140 nm in the lateral plane, a factor of 4–7 greater than the known diameter. Other artefacts ‐ broadening and double images ‐ were observed and ascribed to tip anomalies.High force loadings and slow repetitive scanning resulted in controlled removal of parts of the TMV structure. Accordingly, it was possible to reveal and image the central core channel of the TMV. The precision and resolution of dissection induced by AFM is currently limited by the shape of the tip, having a 40‐nm radius of curvature for standard Si3N4 tips. It is estimated that sharper tips, with a radius of curvature of less than 10 nm, should be able to resolve, non‐destructively, the protein subunits in the non‐contact mode, and selectively remove single subunits in the contact mode.

Olfactory neurons are replaced in the adult olfactory epithelium by division and differentiation of cells close to the basement membrane of the olfactory epithelium. Among the basal cells is a stem cell which undergoes an asymmetric division, giving rise to a new stem cell and a precursor cell. The precursor cell divides again symmetrically and there may be additional symmetrical divisions before the progeny leave the cell cycle to differentiate [1]. We report here the characterization of cell lines created by retroviral insertion of the n-myc proto-oncogene into dividing cells of the adult mouse olfactory epithelium [2]. We have approximately 60 cell lines generated by this procedure, many of them cloned by single cell micromanipulation. These clones are heterogeneous in morphology but they fall into three broad categories: class 1 are mainly flat, epithelial-like cells; class 2 are a mixture of epithelial-like cells, round cells, and bipolar cells with short processes; and class 3 are a mixture of very large epithelial-like cells and bipolar cells with very long processes. Due to the slow growth of the class 3 cell lines and the difficulty of cloning them, we have concentrated our analysis on the class 1 and class 2 cells. Expression of neural and non-neural proteins was studied with a combination of immunocytochemistry, Western analysis, and reverse transcriptase polymerase chain reaction (RT-PCR). All cell lines express vimentin and keratin. Class 1 cell lines failed to express any neural cell markers when grown under our standard growth conditions (10% fetal calf serum, Dulbecco’s modified Eagle’s medium (DMEM), 5% CO2). Class 2 cell lines expressed a variety of neuronal proteins (neurofilament, 13tubulin, MAP-2, NCAM), although not all cell lines expressed all markers. Some clonal cell lines expressed the intermediate filament protein of glial cells (GFAP) in addition to neurofilament.