StAR protein is expressed in both medulla and cortex of the bovine and rat adrenal gland

We have employed polyclonal antibodies to a peptide sequence of bovine steroidogenic acute regulatory (StAR) protein and human placental 3beta-hydroxysteroid dehydrogenase (3beta-HSD) to determine the localisation and distribution of these proteins in rat and bovine adrenal glands. Immunohistochemical staining demonstrated the presence of StAR protein in the zona glomerulosa (ZG), zona fasciculata (ZF), zona reticularis (ZR) and in the medulla of both species. For 3beta-HSD, immunostaining was observed in the ZG, ZF and ZR of the rat adrenal and was absent in the medulla. Immunoblotting experiments showed intense bands for StAR protein (30 kDa, 37 kDa) in the mitochondria of bovine ZG, ZF and medulla and a less intense band (30 kDa) in the microsomes. In rat ZG and ZF/R mitochondria only the 30 kDa protein was present. For 3beta-HSD, an intense band (42 kDa) was found in microsomes and mitochondria of rat and bovine ZG and ZFR. A very faint signal for 3beta-HSD was seen in adrenal medulla. In conclusion, StAR (or a closely related) protein is present throughout the adrenal gland in rat and bovine species in contrast to 3beta-HSD which is confined to the steroidogenic zones. The possible function of StAR protein in the adrenal medulla merits investigation.     

Relationship between prothrombin activation fragment F1.2 and international normalized ratio in patients with atrial fibrillation. Stroke Prevention in Atrial Fibrillation Investigators

BACKGROUND AND PURPOSE: The prothrombin time (expressed as the international normalized ratio [INR]) is the standard method of monitoring warfarin therapy in patients with atrial fibrillation. Prothrombin activation fragment F1.2 provides an index of in vivo thrombin generation and might provide a better index of the effective intensity of anticoagulation. We examined the relationship between F1.2 and INR in patients with atrial fibrillation. METHODS: We measured INR and F1.2 levels in 846 patients with atrial fibrillation participating in the Stroke Prevention in Atrial Fibrillation III study. Two hundred nineteen (26%) were taking aspirin alone, 326 (39%) were taking adjusted-dose warfarin, and 301 (36%) were taking a low fixed dose of warfarin (1 to 3 mg) plus aspirin (combination therapy). F1.2 levels were measured with an enzyme-linked immunosorbent assay. RESULTS: Patients receiving adjusted-dose warfarin or combination therapy had significantly higher INR and significantly lower F1.2 values than those on aspirin alone (P < or = .0001 for each of the four comparisons). F1.2 values (nanomolar) were inversely correlated with INR (F1.2 = -0.1 + 2.3[1/INR]; R2 = .37; P < .0001; simple linear regression). However, significant variability remained. Among patients receiving warfarin, older patients had higher F1.2 values than younger patients after adjustment for INR intensity (P < .001) in the model. There was no difference in the relationship between F1.2 and INR between men and women. CONCLUSIONS: Increasing intensity of anticoagulation, as measured by the INR, is associated with decreasing thrombin generation as measured by the F1.2 level, but significant variability exists in this relationship. Older anticoagulated patients have higher F1.2 values than younger patients at equivalent INR values. The clinical significance of these differences is not clear. F1.2 measurement might provide information regarding anticoagulation intensity in addition to that reflected by the INR.     

Demonstration of discrete secreted and membrane-bound ocular mucins in the dog

Our aims were to separate and characterize secreted canine ocular mucins, and to provide definitive evidence of membrane-bound mucins at the canine ocular surface. Mucus was collected by suction from the ocular surface of normal dogs and dispersed in guanidine hydrochloride and a cocktail of protease inhibitors. Caesium chloride density gradient centrifugation separated secreted mucins from membranes, which were collected from the top of the gradients. Membranes were extracted with octyl glucoside and screened using lectins and anti-mucin antibodies. Gradient fractions containing secreted mucins were constituted into pools on the basis of differential lectin and antibody staining. High molecular weight material from each pool was purified by gel filtration. This material, and the membrane extract, were reduced and alkylated. Vacuum blotting of separated materials after agarose gel electrophoresis was used to compare subunit structure. Density gradient profiles indicated three principal secreted glycoprotein peaks: one staining strongly with anti-mucin antibodies. Gel filtration demonstrated that each contained high molecular weight material. Vacuum blots demonstrated the presence of two secreted glycoproteins with differently sized subunits. On the basis of buoyant density, one of these may be lipid complexed. Membrane extracted material stained with anti-mucin antibodies, and vacuum blotting of this material provided evidence for two membrane-bound components. In conclusion, we have shown that normal canine ocular mucus contains two secreted mucins, each exhibiting different subunit structure; one of these mucins may undergo lipid complexation. Normal canine ocular mucus also contains two membrane-bound mucins: one of which is unique among membrane mucins in showing subunit structure.     

Assessment of childhood phobias

Childhood phobias can be successfully treated using a variety of behavioral strategies, provided there has been a psychometrically sound assessment. Measures are also important for the evaluation of treatment efficacy and the testing of hypotheses generated by new ideas and theories of children's phobias. This paper outlines broad-based assessment procedures used in the evaluation of children's phobias, including the behavioral or problem-focused interview, the diagnostic interview, self-report inventories, caregiver completed instruments, behavioral observations, self-monitoring and physiological assessment. Reflecting recent theoretical and clinical advances in the study of childhood internalizing disorders, we also explore laboratory-based measures and family assessment measures. Particular attention is given to psychometric issues and developmental sensitivity in our discussion of these assessment procedures.     

Functional properties of the acidic and basic subunits of the glycinin (11S) soy protein fraction

The functional properties of low- and high-M.W. (LMW and HMW, respectively) acidic subunits and the basic subunit separated from the 11S soy protein fraction were studied and compared with the functional properties of the 11S fraction. Among the functional properties investigated were solubility, emulsification, and viscosity. The results showed that the LMW acidic subunit had higher solubility than the HMW acidic subunit. Among all the samples, the LMW subunit separated by using β-mercaptoethanol (ME) was the most soluble, with a solubility of 98–100% at a pH of 6–12. The solubility profile of the HMW subunit followed a pattern similar to the solubility of 11S. The lowest solubility was observed around pH values in the range close to the isoelectric point for both the LMW and HMW subunit. The basic subunit was not soluble in the pH range 3–10; however, the solubility increased more than 50% at pH 13 compared to the solubility at pH 10. The emulsification capacity of all subunits was higher than 11S in the following descending order: LMW, basic, HMW, 11S. Emulsification activity and stability of the subunits were greater than those of the 11S samples at room temperature and 95°C. With the exception of the LMW subunit separated with ME, the subunits had a higher viscosity than 11S. The basic subunit separated with sodium bisulfite had the highest viscosity of all the samples tested.     

Cellulose acetate: Interactions with aqueous phenol and a transition temperature of about 20°C

Sorption of aqueous phenol onto powdered cellulose acetate can be described with the Langmuir equation at phenol concentrations below 0.10M. Unacetylated hydroxyl groups are probably the primary sites where phenol hydrogen bonds. As the phenol concentration is increased (> 0.13M), additional hydrogen bonding may occur with the carbonyl oxygens on the primary acetate groups, followed by secondary acetates and the beta glucosidic oxygens. Sorption at higher concentrations shows a negative slope for the Langmuir equation, perhaps caused by crowding and partial blocking of sites. Extrapolations to higher phenolic concentrations using the equation from the negative slope isotherm and plotting N versus °C reveal two lines with a point of intersection at 0.77M. Because of the increase slope above 0.77M, cellulose acetate may be dissolving in phenol. At concentrations below 0.10M, two processes were identified using a Van't Hoff plot (9.33 ± 0.30) × 10³ and (?1.36 ± 0.18) X 10³ J/mol. The initial moisture present in the polymer appears to be an important experimental variable and a transition temperature of 20.1 ± 0.2°C, probably due to polymer swelling, is reported.     

Thermal decomposition behavior of poly[3,3-bis(ethoxymethyl) oxetane] and related polyethers

The thermal decomposition behavior of poly[3,3-bis(ethoxymethyl)oxetane] (polyBEMO) was examined and compared to the decomposition of poly(ethylene oxide) (PEO) and poly(tetramethylene oxide) (polyTHF). Differential scanning calorimetric (DSC) studies as a function of heating rates and at constant temperature as a function of time yielded activation energies of 45–50 kcal/mol, characteristic of polyether decomposition. First-order decomposition kinetics were found. The reaction is endothermic, with a heat of decomposition of 18.6 kcal/mol. Effusion mass spectroscopy on polyBEMO showed major peaks at 112, 140, 168, and 174 amu. A mechanism is proposed in which the thermal scission of the ether bonds in both the polymer chain and in the appendanges initiates the decomposition. The main decomposition reaction for polyBEMO can be written as where the appendages and main chain are cleaved in an unknown order.     

Incorporating a Polyethyleneglycol Linker to Enhance the Hydrophilicity of Mitochondria-Targeted Triphenylphosphonium Constructs

The targeting of bioactive molecules and probes to mitochondria can be achieved by coupling to the lipophilic triphenyl phosphonium (TPP) cation, which accumulates several hundred-fold within mitochondria in response to the mitochondrial membrane potential (Δψ_m). Typically, a simple alkane links the TPP to its “cargo”, increasing overall hydrophobicity. As it would be beneficial to enhance the water solubility of mitochondria-targeted compounds we explored the effects of replacing the alkyl linker with a polyethylene glycol (PEG). We found that the use of PEG led to compounds that were readily taken up by isolated mitochondria and by mitochondria inside cells. Within mitochondria the PEG linker greatly decreased adsorption of the TPP constructs to the matrix-facing face of the mitochondrial inner membrane. These findings will allow the distribution of mitochondria-targeted TPP compounds within mitochondria to be fine-tuned.     

Characterization of Thiol‐Ene Crosslinked PEG Hydrogels

The properties of synthetic hydrogels can be tuned to address the needs of many tissue‐culture applications. This work characterizes the swelling and mechanical properties of thiol‐ene crosslinked PEG hydrogels made with varying prepolymer formulations, demonstrating that hydrogels with a compressive modulus exceeding 600 kPa can be formed. The amount of peptide incorporated into the hydrogel is shown to be proportional to the amount of peptide in the prepolymer solution. Cell attachment and spreading on the surface of the peptide‐functionalized hydrogels is demonstrated. Additionally, a method for bonding distinct layers of cured hydrogels is used to create a microfluidic channel.

     

The Effect of Water Evaporation on Photoacoustic Signals in Transition and Molecular Flow

Evaporation of water changes the response of photoacoustic instruments to light-absorbing particles. Previous calculations of this effect are valid for particles much larger than the mean free path of air. These calculations are extended here to include transition and molecular flow as well as various mass accommodation coefficients for water. For commonly encountered conditions, evaporation can significantly reduce the photoacoustic signal if the mass accommodation coefficient of water on aerosol particles is larger than about 0.01. Unlike the growth of cloud droplets, the photoacoustic signal is very sensitive to changes in the accommodation coefficient between 0.1 and 1. This may provide a way to measure large accommodation coefficients. For a given accommodation coefficient, the change in the photoacoustic signal depends more on absolute than relative humidity. To minimize the effects of evaporation it is better to remove water from the air rather than reduce relative humidity with heating.