Biological freezing and cryofixation

Freezing and freeze fixation are commonly used to achieve ultrastructural and biological preservation. Freezing in biological materials is complex because of their heterogeneous nature—water is unevenly distributed and the various domains are separated by semi-permeable membranes. Processes to be considered include: (1) osmotic gradients leading to redistribution of water, (2) nucleation and uncontrolled growth of ice crystals, (3) recrystallization of nucleated aqueous substrate. To avoid ultrastructural deformation in biological specimens cryofixatives are commonly employed. These are water soluble molecules, able to penetrate cell membranes (e.g. glycerol and dimethylsulphoxide). Interacting strongly with water, ions and bipolymers, they give rise to metabolic and physiological changes which render them useless for X-ray microprobe analytical studies. However, they can enable tissues to survive low temperature storage. Some plants and animals develop in vivo mechanisms which enable them to avoid or tolerate freezing. Alternative means of cryofixation have recently been developed. They rely on non-penetrating polymers of high and specific water binding capacity. These polymers enable the extracellular spaces to be vitrified rather than frozen. Such suppression of ice nuclei enables the cell contents to be maximally subcooled, resulting in the formation of nm dimension ice crystals. Since the polymers have a low osmotic activity and do not penetrate membranes, the interior of the cell is substantially undisturbed. Also hydrophilic polymers used as cryofixatives are physiologically less active than conventional cryoprotectants at equivalent weight concentrations, and their mechanical properties render them useful as matrices for cryosectioning.     

Visual sensitivity of the eye to infrared laser radiation.

The foveal sensitivity to several near-infrared laser wavelengths was measured. It was found that the eye could respond to radiation at wavelengths at least as far as 1064 nm. A continuous 1064 nm laser source appeared red, but a 1060 nm pulsed laser source appeared green, which suggests the presence of harmonic generation in the retina.     

Storage of chicken breast meat in paper coated with different types of hydrophobic agents

Chicken meat is well known for its high nutritional value. Its shelf life is short, due to the pronounced perishability, even under refrigeration. Paper bags are usually used as packaging materials, due to their biodegradability. The main negative aspect concerning paper is its high porosity and low moisture barrier. In this regard, it has to be coated/modified with efficient moisture barrier materials. The storage of chicken breast meat wrapped in four types of paper-based materials currently in use in Romania (RO), Italy (IT), Spain (SP) and Slovenia (SLO) is reported. RO and IT are paper coated with polyethylene (PE) film, SP is waxed paper, and SLO is paper modified with hydrophobized starch. The influence of the coating film on the organoleptical, biochemical and microbiological characteristics of the chicken breast meat stored at 4°C was the aim of the study. The mathematical modelling revealed two aspects: (1) the stability period in which the meat safe for consumption is in the range 1.92–2.18 days in the case of all paper types, and (2) cadaverine, the amine generated by protein degradation, is a less sensitive indicator of the meat alteration than microbial growth. The papers coated with PE (RO and IT samples) and with hydrophobized starch (SLO paper) were packaging materials more efficient that the waxed paper (SP sample). The polyethylene film reduced the conversion of putrescine into spermidine in a higher extent than wax.     

Impact of packaging properties on the physical-chemical-microbiological-sensory characteristics of Ricotta cheese during storage

Ricotta is a highly perishable cheese of which shelf-life is strongly influenced by the properties of packages in which it is stored. Four plastic packages currently in use in Italy (IT), Romania (RO), Slovenia (SLO), and Spain (SP) are characterized in term of structure, barrier, and antimicrobial properties, and their efficiency in the ricotta storage at 4°C is assessed. Differential scanning calorimetry and Fourier-transform infrared spectroscopy analysis indicate that the IT and SP packages are made of low-density polyethylene and the RO and SLO packages consist of polyethylene and polyamide 6. Permeability to the water vapour and oxygen transmission rate vary as follows: IT>SLO > RO > SP and SP > IT>RO > SLO, respectively. The SP package expressed the highest antimicrobial inhibition whilst the IT package the lowest. The maximum amounts of volatile compounds in cheese were reached after 3 days in the IT and SP packages and after 6 days in the SLO and RO packages. The score 2.8 for cheese overall acceptability was exceeded after 7 days in the IT, RO, and SP packages and 6 days in the SLO package. Barrier properties of packages significantly influence the shelf-life of ricotta. Low barrier properties resulted in high level of oxygen and water vapours inside the headspace of packages ultimately favour the microbial development. The simultaneous compliance of microbiological safety level and overall acceptability score gives ricotta a shelf-life of 3 days in the IT, RO, and SP packages and 1 day in the SLO package.     

Development of Long-Wavelength-Emitting Scintillators with Improved Decay Time Characteristics

Liquid and plastic fluors have been prepared which emit in the 550- to 600-nm region. Ternary liquid systems with decay times as short as 1.3 ns at 560 nm and plastic fluors with decay times less than 3 ns at 560 nm are reported. Other liquid and plastic fluors are reported with improved emission characteristics in the region of 600 nm. Conversion efficiences, on a pulse amplitude basis, are generally lower than that of a commercially available 570 nm - 16 ns plastic fluor.     

Enhanced Modeling and Solution of Layered Queueing Networks

Layered queues are a canonical form of extended queueing network for systems with nested multiple resource possession, in which successive depths of nesting define the layers. The model has been applied to most modern distributed systems, which use different kinds of client-server and master-slave relationships, and scales up well. The layered queueing network (LQN) model is described here in a unified fashion, including its many more extensions to match the semantics of sophisticated practical distributed and parallel systems. These include efficient representation of replicated services, parallel and quorum execution, and dependability analysis under failure and reconfiguration. The full LQN model is defined here and its solver is described. A substantial case study to an air traffic control system shows errors (compared to simulation) of a few percent. The LQN model is compared to other models and solutions, and is shown to cover all their features.     

Androgen Reduces Mitochondrial Respiration in Mouse Brown Adipocytes: A Model for Disordered Energy Balance in Polycystic Ovary Syndrome

Polycystic ovary syndrome (PCOS) is a common endocrinopathy that is associated with an adverse metabolic profile including reduced postprandial thermogenesis. Although abnormalities in adipose tissue function have been widely reported in women with PCOS, less is known about direct effects of androgen on white and, particularly, brown adipocytes. The purpose of this study was to investigate the effect of the nonaromatizable androgen dihydrotestosterone (DHT) on (1) lipid accumulation and expression of adipogenic markers in immortalized mouse brown adipose cell lines (IMBATs), (2) mitochondrial respiration in IMBATs, (3) mitochondrial DNA content and gene expression, (4) expression of brown adipose tissue (BAT) markers and thermogenic activation. In addition, we profiled the relative levels of 38 adipokines secreted from BAT explants and looked at androgen effects on adipokine gene expression in both IMBATs and immortalized mouse white adipose (IMWATs) cell lines. Androgen treatment inhibited IMBAT differentiation in a dose-dependent manner, reduced markers of adipogenesis, and attenuated the β-adrenoceptor-stimulated increase in uncoupling protein-1 (UCP1) expression. In explants of mouse interscapular BAT, androgen reduced expression of UCP1, peroxisome proliferator-activated receptor-γ coactivator-1 (PCG-1) and Cidea. Significantly, as well as affecting genes involved in thermogenesis in BAT, androgen treatment reduced mitochondrial respiration in IMBATs, as measured by the Seahorse XF method. The results of this study suggest a role for excess androgen in inhibiting brown adipogenesis, attenuating the activation of thermogenesis and reducing mitochondrial respiration in BAT. Together, these data provide a plausible molecular mechanism that may contribute to reduced postprandial thermogenesis and the tendency to obesity in women with PCOS.     

Development of soluble glasses for biomedical use Part II: The biological response of human osteoblast cell lines to phosphate-based soluble glasses

Soluble glasses are considered to be of potential clinical value in orthopaedic and dental surgery. However, the biological response to these materials is not well understood. To determine the effects of these glasses, two human osteoblast cell lines, MG63 and HOS (TE85), were incubated in vitro in the presence of increasing concentrations of extracts of the glasses. The effects of the extracts on cell growth was measured using the MTT assay and an ELISA assay was used to measure the expression of bone sialoprotein (BSP), osteonectin (ON) and fibronectin (FN), antigens which play a fundamental part in the integrity and function of hard connective tissue. The results showed that the proliferation of the cells was adversely affected only by the more soluble glasses, which also down-regulated the expression of the bone-associated proteins. In contrast, the extract of the glass with the lowest dissolution rate, which contains relatively elevated levels of Ca²⁺, was found to enhance bone cell growth and antigen expression. These findings suggest that the compositions of these glasses at least partly determine the response of cells and thus, that the glasses could be modified to elicit a more optimal biological response and clinical efficacy.     

Specimen coating for high resolution scanning electron microscopy

Thin conducting films, produced by evaporation or soft vacuum sputtering generally show cracks and grain formation, when examined under high resolution scanning electron microscopy (SEM). These artefacts can obscure surface features of coated specimens or cause confusion in the interpretation of micrographs. No such structures have been observed in films produced by ion beam deposition. Ion beam deposition equipment is described in which a cold cathode saddle field ion source, operating at low pressure (15mPa), produces a 2 mm diameter beam of energetic ions (5 keV) and neutrals. With the beam directed onto a target at 30° to glancing incidence, the sputtered material coats the specimens, which are held in a planetary system for good coverage. Conditions favouring fine grain growth are a high nucleation density and low energy transfer to the substrate by thermal conduction or radiation or by particle or photon radiation. These conditions are satisfied by ion beam deposition but evidently not by evaporation or soft vacuum sputtering. With the specimen stationary, sharp shadowing is obtained because the target acts almost as a point source, because of the small diameter of the beam and because there is little scatter at the operating pressure.     

Educational status and health

Some of the problems with the traditional measures of socioeconomic status include (1) the loss of information resulting from combining different factors that have varying associations with health problems; (2) the reverse causal pathway that exists from health and illness to income and occupation; and (3) a number of particular problems with deriving socioeconomic status from census tract information. In contrast there are clear advantages to using educational status as the primary socioeconomic index. A wide variety of literature is reviewed pointing to a strong positive relationship between years of schooling and health. Three models that attempt to account for this association are described. It is suggested that the educational status of patients should be part of their data base.