Room temperature housing results in premature cancellous bone loss in growing female mice: implications for the mouse as a preclinical model for age-related bone loss.

UNLABELLED: Room temperature housing (22 °C) results in premature cancellous bone loss in female mice. The bone loss was prevented by housing mice at thermoneutral temperature (32 °C). Thermogenesis differs markedly between mice and humans and mild cold stress induced by standard room temperature housing may introduce an unrecognized confounding variable into preclinical studies. INTRODUCTION: Female mice are often used as preclinical models for osteoporosis but, in contrast to humans, mice exhibit cancellous bone loss during growth. Mice are routinely housed at room temperature (18-23 °C), a strategy that exaggerates physiological differences in thermoregulation between mice (obligatory daily heterotherms) and humans (homeotherms). The purpose of this investigation was to assess whether housing female mice at thermoneutral (temperature range where the basal rate of energy production is at equilibrium with heat loss) alters bone growth, turnover and microarchitecture. METHODS: Growing (4-week-old) female C57BL/6J and C3H/HeJ mice were housed at either 22 or 32 °C for up to 18 weeks. RESULTS: C57BL/6J mice housed at 22 °C experienced a 62 % cancellous bone loss from the distal femur metaphysis during the interval from 8 to 18 weeks of age and lesser bone loss from the distal femur epiphysis, whereas cancellous and cortical bone mass in 32 °C-housed mice were unchanged or increased. The impact of thermoneutral housing on cancellous bone was not limited to C57BL/6J mice as C3H/HeJ mice exhibited a similar skeletal response. The beneficial effects of thermoneutral housing on cancellous bone were associated with decreased Ucp1 gene expression in brown adipose tissue, increased bone marrow adiposity, higher rates of bone formation, higher expression levels of osteogenic genes and locally decreased bone resorption. CONCLUSIONS: Housing female mice at 22 °C resulted in premature cancellous bone loss. Failure to account for species differences in thermoregulation may seriously confound interpretation of studies utilizing mice as preclinical models for osteoporosis.

Towards a standard for the dynamic measurement of pressure based on laser absorption spectroscopy.

We describe an approach for creating a standard for the dynamic measurement of pressure based on the measurement of fundamental quantum properties of molecular systems. From the linewidth and intensities of ro-vibrational transitions we plan on making an accurate determination of pressure and temperature. The goal is to achieve an absolute uncertainty for time-varying pressure of 5 % with a measurement rate of 100 kHz, which will in the future serve as a method for the traceable calibration of pressure sensors used in transient processes. To illustrate this concept we have used wavelength modulation spectroscopy (WMS), due to inherent advantages over direct absorption spectroscopy, to perform rapid measurements of carbon dioxide in order to determine the pressure. The system records the full lineshape profile of a single ro-vibrational transition of CO2 at a repetition rate of 4 kHz and with a systematic measurement uncertainty of 12 % for the linewidth measurement. A series of pressures were measured at a rate of 400 Hz (10 averages) and from these measurements the linewidth was determined with a relative uncertainty of about 0.5 % on average. The pressures measured using WMS have an average difference of 0.6 % from the absolute pressure measured with a capacitance diaphragm sensor.

Normal bronchial epithelial cell expression of glutathione transferase P1, glutathione transferase M3, and glutathione peroxidase is low in subjects with bronchogenic carcinoma.

Normal bronchial epithelial cells (NBECs) are at risk for damage from inhaled and endogenous oxidative species and from epoxide metabolites of inhaled polycyclic aromatic hydrocarbons. Epidemiological and in vitro data suggest that interindividual variation in this risk may result from variation in NBEC expression of enzymes that inactivate reactive species by conjugating them to glutathione. Quantitative competitive reverse transcription-PCR was used to measure mRNA levels of glutathione transferases (GSTs) and glutathione peroxidases (GSHPxs) in primary NBECs from subjects with or without bronchogenic carcinoma. Mean expression levels (mRNA/10(3) beta-actin mRNA) in NBECs from 23 subjects without bronchogenic carcinoma compared to those from 11 subjects with bronchogenic carcinoma respectively (in parentheses) were: mGST (26.0, 6.11), GSTM3 (0.29, 0.09), combined GSTM1,2,4,5 (0.98, 0.60), GSTT1 (0.84, 0.76), GSTP1 (287, 110), GSHPx (140, 62.1), and GSHPxA (0.43, 0.34). Levels of GSTP1, GSTM3, and GSHPx were significantly (P < 0.05) lower in NBECs from subjects with bronchogenic carcinoma. Further, the gene expression index formed by multiplying the values for mGST x GSTM3 x GSHPx x GSHPxA x GSTP1 had a sensitivity (90%) and specificity (76%) for detecting NBECs from bronchogenic carcinoma subjects that was better than any individual gene. In cultured NBECs derived from eight individuals without bronchogenic carcinoma and incubated under identical conditions such that environmental effects were minimized, the mean level of expression and degree of interindividual variation for each gene evaluated was less than that observed in primary NBECs. Data from these studies support the hypotheses that (a) interindividual variation in risk for bronchogenic carcinoma results in part from interindividual variation in NBEC expression of antioxidant genes; (b) gene expression indices will better identify individuals at risk for bronchogenic carcinoma than individual gene expression values; and (c) both hereditary and environmental exposures contribute to the level of and interindividual variation in gene expression observed in primary NBECs. Many epidemiological studies have been designed to evaluate risk associated with polymorphisms or gene expression levels of putative susceptibility genes based on measurements in surrogate tissues, such as peripheral blood lymphocytes. Based on data presented here, it will be important to include the assessment of NBECs in future studies. Measurement of antioxidant gene expression in NBECs may identify the 5-10% of individuals at risk for bronchogenic carcinoma. Bronchoscopic sampling of NBECs from smokers and ex-smokers then will allow susceptible individuals to be entered into surveillance and/or chemoprevention studies.

Progressive supranuclear palsy: neuropathologic and clinical heterogeneity.

To investigate potential heterogeneity in progressive supranuclear palsy (PSP), we examined 13 patients with neuropathologically diagnosed PSP. The clinical diagnosis of PSP was made in eight of these individuals, whereas probable AD was the primary diagnosis in the remaining five. In addition to PSP neuropathology, seven of the 13 patients (54%) showed concomitant pathologic changes of Alzheimer's disease (AD), Parkinson's disease (PD), or both disorders. These observations indicate that AD and PD changes coexist with PSP neuropathology in a substantive proportion of patients and provide further evidence of clinical and neuropathologic heterogeneity in neurodegenerative disorders. Moreover, our results suggest that PSP may be underdiagnosed and deserves more prominence in the differential diagnosis of dementing illness.

Molecular orientation arrangements in the smectic-C* variant liquid-crystal phases.

Recent experiments have identified three-layer and four-layer distorted helical structures in the smectic liquid-crystal phases Sm C*(FI1) and Sm C*(FI2), respectively. However, no theories have explained the existence of all these phases. A discrete phenomenological model of the free-energy is analyzed and found to predict the stability of distorted three-layer and four-layer structures, as well as simple helical solutions in smectic liquid crystals. A simple physical picture is provided to explain the stability of the phases exhibiting distorted helical structures.

Detailed optical studies of several banana-shaped compounds in liquid crystal B2 phase.

Null-transmission ellipsometry has been performed on free-standing films of one compound in the liquid crystal B2 phase. We have studied films of thickness from 1 to 121 layers. One of the compounds used has an unusually wide 59 K window for the B2 phase. The tilt angle was investigated as a function of temperature and found to be constant over this temperature range to within our resolution of 1 degrees. The one-layer films studied exhibit the same structure as the thicker films, and have helped us to refine the optical model for the B2 phase. For thin films we find that modeling a single smectic layer as two uniaxial layers is a better description of the data than a single biaxial layer, but that for thick films the model used does not effect the simulated result appreciably. Preliminary results also find that the surface layers are less tilted than the interior layers, in contrast to rodlike liquid crystals, which show an enhanced surface tilt.

Surface structures and transitions in the smectic-C* phase of one chiral liquid crystal compound.

Three optical probes have been employed on free-standing films to study surface structure and transitions of the SmC* phase of one liquid crystal compound. While the interior layers show the SmC* structure, the tilt in adjacent surface layers is found to be anticlinic. The number of anticlinic surface layers grows rapidly as the transition to the SmC*(FI2) phase is approached.

Optical studies of the smectic-C(*)(alpha) phase layer structure in free-standing films.

The layer structure of the smectic-C(*)(alpha) phase of one liquid-crystal compound has been acquired from both differential optical reflectivity and ellipsometry measurements in the free-standing film geometry. The data from both techniques display characteristic oscillations as a function of temperature, which can be described by a model for the film consisting of surface anticlinic layers and an interior short-pitched azimuthal helix. These results are consistent with those found previously for another compound. Depolarized reflected light microscopy is used to study the films when the unique features of the aforementioned oscillations occur.

Dramatic effect of an additional CH2 group on the temperature variation of the Sm-C(*)(alpha) short helical pitch.

High-resolution differential optical reflectivity was used to study the temperature evolution of the short helical pitch in the Sm-C(*)(alpha) phase of successive members from two liquid-crystal homologous series. With the addition of one CH2 group, the magnitude and temperature evolution of the pitch change dramatically, and the molecular arrangements between consecutive surface layers found in free-standing films change from being anticlinic to synclinic.

Optical studies of the liquid crystal B2 phase formed by banana-shaped molecules.

Null-transmission ellipsometry and depolarized light microscopy have been performed on free-standing films of three achiral banana-shaped compounds in the B2 phase. Our results support a two-layer unit cell previously proposed to explain the observed antiferroelectricity in thin films and bulk samples. We have studied thicker films than previously reported and have found no deviations in the film structure from the earlier findings. Moreover, we can determine the layer spacing, the molecular tilt from the layer normal, and the three principal indices of refraction in the molecular reference frame.

Nonplanar structure of molecular tilt planes in the surface layers of smectic-A free-standing liquid crystal films.

We present ellipsometric results from thin free-standing films of one chiral liquid crystal compound. In the bulk SmA range with surface-induced molecular tilt, a nonplanar arrangement of the molecular orientations of the tilted surface layers is found under a small applied electric field.

Effect of enantiomeric excess on surface structures and phase sequences in free-standing liquid-crystal films.

We present differential optical reflectivity and null transmission ellipsometry data from several mixtures of MHPBC enantiomers. From this data, we have determined the chiral smectic phase sequence as a function of enantiomeric excess. The data suggest a change in the structure of the smectic-C(*)(alpha) phase and the surface tilt state as the enantiomeric excess is reduced. The results also show that the intermediate phases, smectic C(*)(FI2), smectic C(*)(FI1), and smectic C(*)(alpha), disappear sequentially on reducing the net chirality.

Gelation, oxygen permeability, and mechanical properties of mammalian and fish gelatin films.

The objective of this study was to evaluate the gelation, thermal, mechanical, and oxygen permeability properties of different mammalian, warm- and cold-water fish gelatin solutions and films. Mammalian gelatin solutions had the highest gel set temperatures, followed by warm-water fish and then cold-water fish gelatin solutions. These differences were related to concentrations of imino acids present in each gelatin, with mammalian gelatin having the highest and cold-water fish gelatin having the lowest concentrations. Mammalian and warm-water fish gelatin films contained helical structures, whereas cold-water fish gelatin films were amorphous. This was due to the films being dried at room temperature (23 °C), which was below or near the gelation temperatures of mammalian and warm-water fish gelatin solutions and well above the gelation temperature of cold-water fish gelatin solutions. Tensile strength, percent elongation, and puncture deformation were highest in mammalian gelatin films, followed by warm-water fish gelatin film and then by cold-water fish gelatin films. Oxygen permeability values of cold-water fish gelatin films were significantly lower than those for mammalian gelatin films. These differences were most likely due to higher moisture sorption in mammalian gelatin films, leading to higher oxygen diffusivity.

Analysis of bacterial isolates from cerebrospinal fluid.

We set out to determine the frequency of occurrence of contamination of cerebrospinal fluid with bacteria, seeking also to identify aids to differentiating contaminants from etiologically significant isolates. From 2,091 specimens, 182 bacterial isolates were obtained from 129 patients. Meningitis was the source of 81 isolates (32 patients); contamination yielded 101 isolates (97 patients). The cell counts and protein and glucose concentrations in the cerebrospinal fluid were significantly more often abnormal in specimens from patients with meningitis. Haemophilus influenzae and enteric gram-negative bacilli were usually cause for meningitis, whereas Staphylococcus epidermidis was the most common contaminant. In view of the reported high rate of procedural error in carrying out lumbar puncture, a program aimed at teaching proper technique is recommended to decrease the frequency of false-positive cultures of cerebrospinal fluid.

And a doctor shall lead them....

With managed care reshaping every aspect of health care, physicians need to assume leadership roles more than ever before. This requires skills they don't teach in medical school.