Calpain-induced light scattering by crystallins from three rodent species

The purpose of the present investigation was to compare in vitro light scattering in the soluble proteins from rodent lenses after hydrolysis by the calcium-activated protease, m-calpain (EC 3.4.22.17). Light scattering was measured in solutions of lens proteins from mice, rats, and guinea pigs after activation of endogenous m-calpain or after addition of purified m-calpain. We found for the first time that, in addition to rat, crystallins from another rodent lens, young mouse, were susceptible to calpain-induced light scattering. As in rats, aging of mouse lens prevented calpain-induced light scattering. Although crystallins from guinea pig lens were also partially hydrolysed by calpain, appreciable light scattering did not occur. Limited proteolysis may cause common changes in the biophysical properties of mouse and rat crystallins to decrease their solubility. Discovery of the nature of these biophysical changes may help our understanding as to why crystallins precipitate under cataractous conditions.     

Mice lacking serum paraoxonase are susceptible to organophosphate toxicity and atherosclerosis

Serum paraoxonase (PON1) is an esterase that is associated with high-density lipoproteins (HDLs) in the plasma; it is involved in the detoxification of organophosphate insecticides such as parathion and chlorpyrifos. PON1 may also confer protection against coronary artery disease by destroying pro-inflammatory oxidized lipids present in oxidized low-density lipoproteins (LDLs). To study the role of PON1 in vivo, we created PON1-knockout mice by gene targeting. Compared with their wild-type littermates, PON1-deficient mice were extremely sensitive to the toxic effects of chlorpyrifos oxon, the activated form of chlorpyrifos, and were more sensitive to chlorpyrifos itself. HDLs isolated from PON1-deficient mice were unable to prevent LDL oxidation in a co-cultured cell model of the artery wall, and both HDLs and LDLs isolated from PON1-knockout mice were more susceptible to oxidation by co-cultured cells than the lipoproteins from wild-type littermates. When fed on a high-fat, high-cholesterol diet, PON1-null mice were more susceptible to atherosclerosis than their wild-type littermates.     

Comparison of methods to assess visual impairment from glare and light scattering with posterior capsule opacification

PURPOSE: To compare 2 glare tests to determine their relative usefulness in the assessment of posterior capsule opacification (PCO) and to evaluate the potential benefits of combined visual, acuity, contrast sensitivity, and glare testing. SETTING: Teaching hospital ophthalmology department. METHODS: Sixteen patients had glare, visual acuity, and contrast sensitivity testing before and after neodymium:YAG (Nd:YAG) capsulotomy. Results with the Brightness Acuity Tester (BAT, Mentor), which measures disability glare, and the Straylightmeter (Foundation for Eye Research, The Netherlands), which quantifies forward scatter by direct compensation techniques, were compared. The correlation between glare, ETDRS visual acuity, and Pelli-Robson contrast sensitivity was determined. RESULTS: Pretreatment visual acuity was significantly correlated with contrast sensitivity (P < .01). However, visual acuity and contrast sensitivity were poorly correlated with both the BAT and Straylightmeter (P > .05), indicating that visual acuity is predictive of contrast sensitivity but a poor predictor of glare. Glare was significantly improved (Straylightmeter, P < .0001; BAT, P < .05) following capsulotomy. While the Straylightmeter consistently measured precapsulotomy forward scatter that improved with treatment, corresponding BAT disability glare was unmeasurable in 18.8% of patients with PCO, as their visual acuities improved rather than deteriorated with glare testing. CONCLUSIONS: Glare testing provided more information than contrast sensitivity when combined with visual acuity in the evaluation of PCO. Glare related to PCO is better assessed using the Straylightmeter because the BAT may yield aberrant disability glare results.     

Testing whether correlation matrices are different from each other.

Developmental psychologists use correlation matrices both as tools for summarizing the relations among sets of variables and as input to multivariate statistical procedures. This article reviews methods for testing whether 2 or more correlation matrices are different from each other. Methods are illustrated for evaluating the similarity of 2 independent correlation matrices, such as those obtained from boys and girls, and 2 dependent correlation matrices, such as those obtained in longitudinal research by using multiple measures. Applications of the models to data from the published developmental literature are provided. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Infants entering foster care compared to other infants using birth status indicators

OBJECTIVE: Infants comprise nearly one fourth of all entries to foster care. Linkage of administrative birth and placement data can provide information about these infants that may otherwise be unavailable or difficult to obtain. METHOD: Statewide birth records and foster care placement histories were linked via probability matching. Logit regression was used to compare 26,460 maltreated infants who entered foster care between 1989 and 1994 with a random sample of 68,401 other infants born during that time frame. RESULTS: Infants in care were more than twice as likely to have single parents and be born with low birthweight, and twice as likely to have been born with a birth abnormality as other infants, controlling for other factors. The largest difference was in the eightfold increased likelihood for mothers of infants in care to have had no prenatal care. Infants in care were nearly three times as likely to be born into larger families (third or greater live births to the mother). Mothers of infants in care were more than twice as likely to be African American compared to White than mothers of other infants, while Hispanic and Other ethnic groups were underrepresented in the group of infants in care. Foreign born mothers, especially Hispanic women, were much less likely to have infants in care than they were to have children in the other group. CONCLUSIONS: Administrative datasets, while often limited in the number of variables they include and scope of their information, can be a valuable tool when used to understand demographics and frame questions for future research. Infants who enter foster care differ in substantial ways from other children. These findings have important implications for future research aimed toward targeting of child welfare services and supports.     

Comparative immunological studies between canine prolactin and prolactin from other species

Canine pituitary extracts contained material which was capable of inhibiting the binding between antisera to different prolactins and labelled non-canine prolactins. Maximum inhibition of binding was observed in the system consisting of antisera to ovine prolactin and labelled rat prolactin. This system was used to investigate the immunological activity of canine pituitary extracts and partially purified fractions. A rabbit antiserum which was raised against a canine pituitary extract and had significant immunological activity in the above system, together with labelled rat prolactin, was used to develop an assay capable of detecting material in canine pituitary extracts. This material gave parallel dose-response curves to a canine prolactin-rich fraction, D864C and purified ovine and bovine prolactin.     

Isolation of adenoviruses and reoviruses from avian species other than domestic fowl

Adenoviruses and reoviruses were isolated from pigeons and mallard ducks. In addition, adenoviruses were isolated from budgerigars and a bantam and a reovirus was isolated from a turkey. Primary identification of these viruses was by electron-microscope examination. It was further possible to assign the 4 adenoviruses to recognized fowl serotypes, and the reoviruses shared a common antigen with fowl reoviruses. These viruses were isolated from a variety of clinical conditions.     

Microsporidia are related to Fungi: evidence from the largest subunit of RNA polymerase II and other proteins

We have determined complete gene sequences encoding the largest subunit of the RNA polymerase II (RBP1) from two Microsporidia, Vairimorpha necatrix and Nosema locustae. Phylogenetic analyses of these and other RPB1 sequences strongly support the notion that Microsporidia are not early-diverging eukaryotes but instead are specifically related to Fungi. Our reexamination of elongation factors EF-1alpha and EF-2 sequence data that had previously been taken as support for an early (Archezoan) divergence of these amitochondriate protists show such support to be weak and likely caused by artifacts in phylogenetic analyses. These EF data sets are, in fact, not inconsistent with a Microsporidia + Fungi relationship. In addition, we show that none of these proteins strongly support a deep divergence of Parabasalia and Metamonada, the other amitochondriate protist groups currently thought to compose early branches. Thus, the phylogenetic placement among eukaryotes for these protist taxa is in need of further critical examination.     

The effects of a high fat diet on leptin mRNA, serum leptin and the response to leptin are not altered in a rat strain susceptible to high fat diet-induced obesity

Osborne-Mendel (OM) and S5B/Pl rats differ in their sensitivity to develop obesity when fed a high fat (HF) diet; OM rats become obese, whereas S5B/Pl rats remain thin. We have investigated the possibilities that either an impaired leptin response or resistance to leptin action underlies the sensitivity to this form of obesity in OM rats. In Experiment 1, OM and S5B/Pl rats fed a nonpurified diet were killed at d 0 or were fed either a HF (56% fat energy) or a low fat (LF, 10% fat energy) diet for 2 or 7 d. The HF diet increased serum leptin significantly by d 2 to levels that were similar in both rat strains. At 7 d, leptin levels were lower than at d 2 but remained higher than levels in the d 0 control groups. The leptin mRNA:18S RNA ratio in epididymal adipose tissue increased to higher levels in HF-fed OM rats than in S5B/Pl rats fed that diet. However, although the LF diet had no effect in S5B/Pl rats, it increased leptin mRNA levels in epididymal adipose tissue of OM rats compared with the controls fed the nonpurified diet. In Experiment 2, OM and S5B/Pl rats were fed HF or LF diets for 5 wk. At that time, their feeding response to a range of leptin doses (0, 1, 5 or 10 microgram) given intracerebroventricularly was tested after overnight food deprivation. There was a similar dose-dependent reduction in energy intake in response to leptin in both OM and S5B/Pl rats. These responses were independent of the diet. The data suggest that the susceptibility of OM rats to HF diet-induced obesity is not related to either a loss of central sensitivity to leptin or a failure to enhance leptin production acutely, although the failure to maintain chronically increased levels of serum leptin could contribute to the obesity.     

Fiber-type caffeine sensitivities in skinned muscle fibers from humans susceptible to malignant hyperthermia

BACKGROUND: The response to contracture tests may depend upon the relative proportion of muscle fiber types within the muscle specimen. To determine whether a difference in fiber-type caffeine sensitivities exists between malignant hyperthermia susceptible (MHS) and malignant hyperthermia-nonsusceptible (MHN) skeletal muscle, we compared the fiber-type caffeine sensitivities in chemically skinned muscle fibers dissected from vastus lateralis muscle from 15 MHS and 16 MHN patients. METHODS: Muscle fiber type was determined in each fiber by the difference in strontium-induced tension measurements and in 36 fibers, after contracture testing, by ATPase enzyme histochemistry. Caffeine sensitivity was defined as the threshold concentration inducing more than 10% of the maximal tension obtained with a calcium 1.6 x 10(-2) mM solution. RESULTS: Significant difference in the mean (+/- SD) caffeine sensitivity was found between type I MHS fibers (2.63 +/- 0.85 mM) versus type II MHS fibers (3.47 +/- 1.2 mM) and between type I MHN fibers (5.89 +/- 1.8 mM) versus type II MHN fibers (10.46 +/- 2.6 mM). The mean (+/- SD) caffeine sensitivities for a given muscle fiber type (I or II) were different between groups of MHS and MHN patients. Both type I and II MHS fibers had significantly lower caffeine sensitivities, and this increase in caffeine sensitivity was significantly smaller in type I than in type II fiber. CONCLUSIONS: The current study indicates that a truly MHS patient cannot have a false-negative result solely related to abnormal type II fibers contained in a given muscle strip. Although the occurrence of a very high proportion of type I fibers in MHN human muscle could result in a false-positive contracture outcome, such an occurrence is expected to be rare.     

Hydrodynamic properties of nitrogenase--the MoFe protein from Azotobacter vinelandii studied by dynamic light scattering and hydrodynamic modelling

Trichloroethylene (TRI) is an industrial solvent with a history of use in anesthesia, and is a common groundwater contaminant. Cytochrome P450 (CYP)-dependent metabolism of TRI produces chloral hydrate (CH) and is rate limiting in the ultimate production of trichloro- and/or dichloroacetic acid from TRI. Exposure of rodents to TRI results in lung and liver tumors (mice) and nephrotoxicity (rats). The toxicity is exacerbated by pretreatment of mice with CYP inducers. We report significant variability in TRI metabolism in a sample of 23 human hepatic microsomal samples and demonstrate the dependence of TRI metabolism on CYP2E1. K(m) values in this limited sample population are not normally distributed. We have correlated microsomal CH formation with the activity toward routine CYP2E1 substrates and with immunologically detectable CYP2E1 protein. Further, TRI metabolism in microsomes from lymphoblastoid cell lines expressing CYP2E1, CYP1A1, CYP1A2, or CYP3A4 indicated minimal involvement of the latter forms, with CYP2E1 catalyzing more than 60% of total microsomal TRI metabolism. These results indicate that humans are not uniform in their capacity for CYP-dependent metabolism of TRI and increased CYP2E1 activity may increase susceptibility to TRI-induced toxicity in the human.     

HTLV-Is in Argentina are phylogenetically similar to those of other South American countries, but different from HTLV-Is in Africa

We present a method for obtaining the line spread function (LSF) of any radiation detector from measured data. The problem of finding a LSF is essentially a discrete deconvolution from known values of the input (Monte Carlo generated data) and the output (measured data) which can be put into matrix form. We applied the total least squares (TLS) method which is particularly useful when there are errors in both the input and output data. Results from computer simulation as well as from actual data are shown. In a practical application, however, our technique is currently limited by the ability of the Monte Carlo data to simulate correctly the inherent data from the head of the linear accelerator (linac). To overcome this difficulty we have solved by deconvolution and TLS for a more realistic inherent beam profile of our linac using the information from both profile data as measured with film and the film densitometer response function. The LSF of the densitometer was estimated with a simple method of direct measurement of a slit image and a full width at half maximum (FWHM) of 0.997 mm was recorded. Additionally, using the knowledge of this realistic inherent profile of the linac, a blurring function representing the finite source size effect missing in our current Monte Carlo profile simulation was determined. Finally, with the realistic inherent beam profile we have applied the deconvolution and TLS method to find a LSF for the Markus chamber and found a resulting FWHM of 5.39 mm. The TLS approach for deconvolving can find a useful application for both finding the LSF and correcting for the detector size effect once its LSF is known. This type of correction is required when a high spatial resolution is needed (e.g., in small field off-axis measurements). Convolved and measured profiles are also presented to illustrate the effect of the blurring due to different LSFs.     

Discrimination of Burkholderia gladioli from other Burkholderia species detectable in cystic fibrosis patients by PCR

A procedure for molecular identification of Burkholderia gladioli is described. Specific 16S and 23S rRNA gene signature sequences were defined as primers for PCR. The method allows rapid and specific discrimination of B. gladioli from related species (B. cepacia, B. multivorans, B. vietnamiensis, B. mallei, B. pseudomallei, Ralstonia pickettii, and R. eutropha) and should contribute to the clarification of its role as a human pathogen, e.g., in cystic fibrosis.