Allograft outcomes in outbred mice.

Inbreeding depression and lack of genetic diversity in inbred mice could mask unappreciated causes of graft failure or remove barriers to tolerance induction. To test these possibilities, we performed heart transplantation between outbred or inbred mice. Unlike untreated inbred mice in which all allografts were rejected acutely (6-16 days posttransplantation), untreated outbred mice had heterogeneous outcomes, with grafts failing early (<4 days posttransplantation), acutely (6-24 days) or undergoing chronic rejection (>75 days). Blocking T cell costimulation induced long-term graft acceptance in both inbred and outbred mice, but did not prevent the early graft failure observed in the latter. Further investigation of this early phenotype established that it is dependent on the donor, and not the recipient, being outbred and that it is characterized by hemorrhagic necrosis and neutrophilic vasculitis in the graft without preformed, high titer antidonor antibodies in the recipient. Complement or neutrophil depletion prevented early failure of outbred grafts, whereas transplanting CD73-deficient inbred hearts, which are highly susceptible to ischemia-reperfusion injury, recapitulated the early phenotype. Therefore, outbred mice could provide broader insight into donor and recipient determinants of allograft outcomes but their hybrid vigor and genetic diversity do not constitute a uniform barrier to tolerance induction.

The confounding effect of response amplitude on MVPA performance measures.

Multi-voxel pattern analysis (MVPA) is proving very powerful in the analysis of fMRI timeseries data, yielding surprising sensitivity, in many different contexts, to the response characteristics of neurons in a given brain region. However, MVPA yields a metric (classification performance) that does not readily lend itself to quantitative comparisons across experimental conditions, brain regions or people. This is because performance is influenced by a number of factors other than the sensitivity of neurons to the experimental manipulation. One such factor that varies widely but has been largely ignored in MVPA studies is the amplitude of the response being decoded. In a noisy system, it is expected that measured classification performance will decline with declining response amplitude, even if the underlying neuronal specificity is constant. We document the relationship between response amplitude and classification performance in the context of orientation decoding in the visual cortex. Flickering sine gratings were presented at each of two orthogonal orientations in a block design (multivariate experiment) or an event-related design (univariate experiment). Response amplitude was manipulated by varying stimulus contrast. Orientation classification performance in retinotopically defined occipital area V1 increased approximately linearly with the logarithm of stimulus contrast. As expected, univariate response amplitude also increased with contrast. Similar results were obtained in V2, V3 and V3A. Plotting classification performance against response amplitude gave a function with a compressive non-linearity that was well fit by a power function. Knowledge of this function potentially allows adjustment of classification performance to take account of the effect of response size, making comparisons across brain areas, categories or people more meaningful.

Type I interferons are not critical for skin allograft rejection or the generation of donor-specific CD8+ memory T cells.

Type I interferons (IFN-I) link innate to adaptive immunity in microbial infection, autoimmune disease and tumor immunity. It is not known whether IFN-I have an equally central role in alloimmunity. Here we tested this possibility by studying skin allograft survival and donor-specific CD8+ T-cell responses in mice that lack the IFN-I receptor (IFN-IR-/-). We found that IFN-IR-/- mice reject fully allogeneic wild-type skin grafts at the same rate as wild-type recipients. Similarly, allograft rejection was not delayed if IFN-IR-/- male skin was transplanted to syngeneic IFN-IR-/- female mice. Quantitation of the male (H-Y)-specific CD8+ T-cell response in these mice revealed normal generation of donor-specific CD8+ effector T cells but fourfold reduction in CD8+ memory T cells. Memory CD8+ T cells generated in the absence of IFN-IR had normal phenotype and recall function, assessed by ex vivo cytokine production and the ability of IFN-IR-/- mice to mount second set rejection. Finally, these memory T cells were maintained at a constant number despite their inability to respond to IFN-1. Our findings indicate that IFN-I cytokines are not critical for acute allograft rejection or for the expansion and differentiation of donor-specific CD8+ T cells into long-lived, functional memory T cells.

Mathematical models for conventional and microwave thermal deactivation of Enterococcus faecalis, Staphylococcus aureus and Escherichia coli.

Temperature dependencies of survival fecal coliforms such as Enterococcus faecalis, Staphylococcus aureus and Escherichia coli in water were investigated between 25-65 degrees C. Measured dependencies had "bell" shaped form with maximum bacterial viability at 35-45 degrees C. The rates of growth and decay of bacterial viability depend on specific forms of bacteria. At temperatures of 60-65 degrees C the number of viable bacteria decreased in one hundred times in comparison with the maximum value. Similar "bell" shape forms were found for dependencies between bacterial viability and time of microwave (dielectric) heating of water. The dependencies had maximum value at 1-2 min of microwave heating. Then, the number of viable bacteria decreased, and at 4-5 min of microwave heating, became insignificantly small. The proposed mathematical models for conventional and microwave heating took into account "growth" and "death" factors of bacteria, and had forms of second degree polynomial functions. The results showed good relationships (with coefficient correlation 0.84-0.99) between the proposed mathematical models and experimental data for both conventional and microwave heating.

Mathematical model of manganese ion catalyzed microwave deactivation of Enterococcus faecalis, Staphylococcus aureus and Escherichia coli.

Enterococcus faecalis, Staphylococcus aureus and Escherichia coli survival was investigated using microwave irradiation (power 130 W) both in a water control and in the presence of a 1 microM manganese ion solution. Measured survival dependencies had "bell" shape form with maximum bacterial viability between 1-2 min of microwave heating. Additional heating revealed bacteria survival decreasing up to 3 min of microwave heating when viability became insignificantly small. The total deactivation time of bacteria in the presence of manganese ions was significantly smaller then that of bacteria irradiated in the microwave without manganese ions present (4-5 min). One possible explanation for the rapid reduction of bacterial survival during microwave irradiation in the presence of manganese ions is that increasing manganese ion penetration into bacteria along with microwave irradiation related to an increase of kinetic energy of ions, and damaging of bacteria by metal ions. The proposed mathematical model for microwave heating took into account "growth" and "death" factors of bacteria. It assumes that rates of bacterial growth and decay are linear functions of water temperature, and rate of bacterial decay that relates with metal concentration into water is also linear, which influenced the differential equation for the dependence between number of survival bacteria and temperature water. By using proportionality between the time of microwave heating and water temperature we derived the differential equation, between bacterial viability and time of microwave irradiation which was used as mathematical model for microwave heating in the presence of metal ions. This model had forms of second-degree polynomial functions. We received good relationships (with coefficient of correlation 0.92-0.99) between proposed mathematical model and experimental data for all bacterial deactivation.

In vitro reconstitution of legumin (11S) mRNA and binding proteins as related to post-transcriptional regulation of protein synthesis in developing alfalfa embryos.

There is undetectable transcription of 11S storage protein (medicagin) mRNA by nuclei isolated from pre-cotyledonary-stage somatic embryos of alfalfa (Medicago sativa L). However, this message exists at steady-state levels in the embryos at this stage of development without concomitant synthesis of the storage protein. At the pre-cotyledonary stage, therefore, the transcriptional rate for 11S mRNA is low; what message is transcribed is sequestered in the form of mRNP complexes and is not recruited into polysomes in vivo (33). Both transcription (in vivo and in isolated nuclei) and translation of the 11S mRNA are evident at the onset of cotyledon development in somatic and zygotic embryos, reaching a maximum during expansion of the cotyledons and then declining as the embryos mature. Pre-cotyledonary-stage somatic embryos which do not utilize the 11S-mRNA in polysomes lack certain mRNA-binding proteins (32, 36 and 38 kD) which are present at later stages of development. These mRNA-binding proteins may be responsible for the initiation of large polysome formation since they were exclusively present in the translational extracts of cotyledonary somatic and zygotic embryos in which there was no repression of storage protein synthesis. In contrast, the pre-cotyledonary somatic embryos contained a different set of 11S-mRNA-binding proteins (28, 50, 55, and 62 kD) whose presence in the cotyledonary stage embryos was very rare or non-existent; these could be responsible for preventing translation.

Effect of Cnidoscolus aconitifolius leaf extract on the blood glucose and insulin levels of inbred type 2 diabetic mice.

The effects of Cnidoscolus aconitifolius (CA) leaf extract and chlorpropamide on blood glucose and insulin levels in the inbred type 2 diabetic mice are reported. After treatment with CA, the glucose levels were measured at 0 and 2-hour intervals in experimental groups and controls. Group I received no treatment and served as control; Group II was the reference and it received chlorpropamide; Groups I-III were moderately diabetic, 100-300 mg/dL blood glucose levels while Group IV were severely diabetic (> 300 mg/dL). Groups III and IV received CA and served as test groups. There was no significant difference between the blood glucose levels at 0 and 2 hours for the control group, (P>0.23) but there were statistically significant differences for Group II (P<0.0002); Group III (P<0.002) and Group IV (P<0.0001). For moderately diabetic mice, CA and chlorpropamide decreased the glucose levels by 25.6% and 16.3% respectively while for the severely diabetic mice CA decreased the blood glucose by 43.7%. It is proposed that CA has an insulinogenic property that possibly stimulated dormant beta-cells to secrete insulin. The histopathology of several organs in the treated animals was found to differ from the expected. The islets of Langerhans for example were found to be preserved in the time frame examined. Also the liver and kidney were found to display milder pathology in the treated groups.

Teaching teamwork during the Neonatal Resuscitation Program: a randomized trial.

OBJECTIVE: To add a team training and human error curriculum to the Neonatal Resuscitation Program (NRP) and measure its effect on teamwork. We hypothesized that teams that received the new course would exhibit more teamwork behaviors than those in the standard NRP course. STUDY DESIGN: Interns were randomized to receive NRP with team training or standard NRP, then video recorded when they performed simulated resuscitations at the end of the day-long course. Outcomes were assessed by observers blinded to study arm allocation and included the frequency or duration of six team behaviors: inquiry, information sharing, assertion, evaluation of plans, workload management and vigilance. RESULT: The interns in the NRP with team training group exhibited more frequent team behaviors (number of episodes per minute (95% CI)) than interns in the control group: information sharing 1.06 (0.24, 1.17) vs 0.13 (0.00, 0.43); inquiry 0.35 (0.11, 0.42) vs 0.09 (0.00, 0.10); assertion 1.80 (1.21, 2.25) vs 0.64 (0.26, 0.91); and any team behavior 3.34 (2.26, 4.11) vs 1.03 (0.48, 1.30) (P-values <0.008 for all comparisons). Vigilance and workload management were practiced throughout the entire simulated code by nearly all the teams in the NRP with team training group (100% for vigilance and 88% for workload management) vs only 53 and 20% of the teams in the standard NRP. No difference was detected in the frequency of evaluation of plans. CONCLUSION: Compared with the standard NRP, NRP with a teamwork and human error curriculum led interns to exhibit more team behaviors during simulated resuscitations.

Regulation of acetohydroxy acid synthase activities in Escherichia coli K-12 by small metabolites.

The effects of several metabolites (indole acetic acid, imidazole acetic acid and indole) on acetohydroxy acid synthase activities have been examined in both cya+ and cya- strains. Specifically, indole acetic acid caused an increase in the rate of acetohydroxy acid synthase synthesis under both in vivo and in vitro conditions. Taken together, these data suggest that small metabolites, other than cAMP, can alter acetohydroxy acid synthase gene expression.

Growth dynamics of the developing lateral geniculate nucleus.

Segregated binocular maps in the lateral geniculate nucleus (LGN) develop from stages where they initially completely overlap. Here, we show that segregation occurs at different rates across the depth of the nucleus and that the volume of the ipsilateral projection does not decrease significantly during this period, rather LGN volume expands markedly and its shape changes. Hence, we have examined the differential growth of the ferret LGN during the process of segregation by using novel shape modelling techniques. These have facilitated quantification of its three-dimensional structure at successive developmental stages as well as the definition of growth vectors which illustrate shape change. This has been undertaken in direct representations of the LGN and those normalised for size and orientation. Spatiotemporal aspects of shape change have then been compared with different measurements of its cellular population. Initial stages of segregation are associated with a large expansion of the rostrocaudal axis of the nucleus along which segregation takes place, and an expansion of caudal regions that will eventually contain the binocular representation. Later stages are associated with dorsoventral expansions and a consolidation of the rostrocaudal axis. The pace of shape change peaks toward the end of the period of segregation when the nucleus has adopted approximately 50% of its adult shape. After segregation, nuclear growth is mainly isotropic. The mature shape of the nucleus is achieved before it reaches its full size and while cell density and cell sizes are still changing.

A clinical evaluation of xenon enhancement for computed tomography.

Xenon increases the attenuation coefficients of tissues in which it dissolves and, therefore, enhances lipid-rich material. The authors evaluated xenon enhancement for computed tomography. Patients and normal subjects were given xenon in subanesthetic concentrations and were scanned in a large aperture CT scanner. Patients tolerated 30-50% inhaled xenon well. Cerebral tissues and lung were enhanced by this concentration of xenon. Xenon inhalation in subanesthetic concentrations will be a useful technique for characterizing abnormal lipid tissues and for estimating cerebral blood flow.

Quantification of contrast in clinical MR brain imaging at high magnetic field.

The relative contrast between two tissues in a magnetic resonance (MR) image is shown to be quantifiable for any combination of pulse timing parameters, provided the intrinsic parameters are known. Based on multiple inversion-recovery and spin echo images, a region-of-interest T1, T2 and density analysis was conducted at 1.4T in selected patients with diagnosed neuropathology for various brain tissues. The resulting tissue parameters subsequently served to calculate the contrast-to-noise (C/N) ratio for typical tissue interfaces as a function of the operator-variable pulse timing parameters and the data were compared with the images. Although such calculations may be useful as a protocol selection aid, it is obvious that an optimized pulse protocol can only be established for a single tissue interface. The data also reveal that a T2-discriminating pulse sequence like Carr-Purcell-Meiboom-Gill with long repetition time, generally advocated as clinically most effective, may not always be ideal.

Model studies toward the total synthesis of the Lycopodium alkaloid spirolucidine.

[reaction: see text] A strategy for the synthesis of the spirocyclic core of spirolucidine was explored through a model study. The diene 4a was prepared and photolyzed to give the desired [2 + 2] photoadduct 17 containing the correct relative stereochemistry corresponding to spirolucidine.

Pneumoperitoneum does not potentiate the nephrotoxicity of aminoglycosides in rats.

OBJECTIVES: Pneumoperitoneum is associated with transient renal dysfunction. To our knowledge, the safety of administering nephrotoxins such as aminoglycosides during pneumoperitoneum has not been studied. Our hypothesis was that pneumoperitoneum potentiates the nephrotoxicity of aminoglycosides. METHODS: From 29 rats we obtained preprocedure 24-hour urine collections. In the pneumoperitoneum group (n = 7), carbon dioxide was insufflated intra-abdominally at 15 mm Hg pressure for 2 hours. In the gentamicin group (n = 7), 10 mg/kg gentamicin was administered intravenously. In the combined pneumoperitoneum/gentamicin group (n = 8), the same dose of gentamicin was administered 10 minutes before pneumoperitoneum. Sham rats (n = 7) received anesthesia only. Urine was collected for the 24 hours after the procedure, and 1 week later blood for creatinine determination and final 24-hour urine collections were obtained. All urine samples were assayed for creatinine and N-acetyl-beta-glucosaminidase (NAG). RESULTS: Only the gentamicin and combined pneumoperitoneum/gentamicin groups presented day 1 values for NAG excretion that were significantly greater than same day sham or paired preprocedure values; the rest of the urinary creatinine and NAG day 1 levels and all the day 7 levels were not significantly different from same day sham or paired preprocedure levels. Day 7 serum creatinine and creatinine clearance did not differ significantly among the groups. CONCLUSIONS: We found that intravenous gentamicin transiently increased urinary excretion of NAG in rats, which resolved within 1 week. Pneumoperitoneum for 2 hours at 15 mm Hg did not increase urinary NAG, either alone or in gentamicin-treated rats. Moreover, our data are sufficient to refute with 95% certainty the possibility that gentamicin plus pneumoperitoneum decreases creatinine clearance more than approximately 60%. These results do not support the hypothesis that pneumoperitoneum potentiates the nephrotoxicity of aminoglycosides.

Microcracks in dental porcelain and their behavior during multiple firing.

Dental porcelains rely on the high-thermal-expansion mineral leucite to elevate their bulk thermal expansion to levels compatible with dental PFM alloys. The microcracks that form around these leucite particles when cooled during porcelain manufacture are a potential source of change in bulk porcelain thermal expansion during fabrication of porcelain-fused-to-metal crowns and bridges. The purpose of the present study was to determine whether multiple firings of commercial dental porcelains could produce changes in microcrack density. Specimens of six commercial porcelains and the "Component No. 1" of the Weinstein patent were fabricated and subjected to 1, 2, 4, 8, and 16 firings. The microcrack densities were determined by quantitative stereology, whereby intersections of microcracks were counted with a test grid. The microcrack data were subjected to linear regression analysis and analysis of variance. The microcrack densities of four of the six porcelains and the Component No. 1 frit were not significantly affected by the number of firings (p > 0.05). One porcelain exhibited a weak but highly significant positive correlation between microcrack density and multiple firings (r2 = 0.24, p = 0.0003), while the remaining porcelain exhibited a weak but statistically significant negative correlation between microcrack density and multiple firings (r2 = 0.15, p = 0.006). The results of this study indicate that even for porcelains that exhibit a measurable change in microcrack density as a function of multiple firings, the magnitude of the increase or decrease in microcrack density after several firings is sufficiently small to cause only negligible shifts in porcelain bulk thermal expansion.

High-temperature X-ray diffraction measurement of sanidine thermal expansion.

Dental porcelains that are designed to be fused to PFM (porcelain-fused-to-metal) alloys are formulated by their manufacturers to be closely matched in thermal expansion to these alloys. The high thermal expansion of the mineral leucite has been exploited to regulate porcelain expansion. Leucite, however, has been observed to convert to the sanidine polymorph of feldspar during certain heat treatments within the normal firing range of dental porcelain. The effects of this conversion on porcelain thermal expansion and porcelain-metal thermal compatibility have been uncertain, due to the paucity of published data on the thermal expansion of sanidine. The purpose of this study was to measure the thermal expansion of sanidine by high-temperature X-ray diffraction over the temperature range in which thermal mismatch stresses can develop in porcelain-fused-to-metal restorations, i.e., from room temperature to 700 degrees C. The lattice parameters a, b, c, and beta were determined from the d-spacings and hkl values of multiple reflections by means of a least-squares iteration. The dependence of each lattice parameter on temperature was determined via analysis of variance, and the coefficient of thermal expansion, alpha, was obtained from this analysis. The lattice parameters of sanidine at room temperature were determined to be: a = 0.8524 +/- 0.0015 nm, b = 1.3020 +/- 0.0004 nm, c = 0.7165 +/- 0.0002 nm, and beta = 116.02 degrees +/- 0.01 degree (mean +/- 95% confidence interval). The linear thermal expansion coefficient, a, over the range from room temperature to 700 degrees C was determined to be 4.1 x 10(-6) K(-1) +/- 0.6 x 10(6) K(-1) (mean +/- 95% confidence interval). Because the coefficient of thermal expansion for sanidine is substantially lower than that of leucite (the effective linear thermal coefficient of thermal expansion of leucite over the range of 25 degrees to 700 degrees C is 28 x 10(-6) K(-1)), the conversion of leucite to sanidine during porcelain heat treatments would produce a detrimental lowering of the porcelain thermal expansion.

MR imaging of prolactin-secreting microadenomas.

Eight proven and three presumed prolactin-secreting microadenomas were studied with magnetic resonance (MR) imaging. All 11 cases had CT evidence of a tumor. Technical factors for MR included use of 1.3-1.5 T MR systems, 3 mm slice thickness, short repetition time (TR) (T1-weighted) and long TR (T2-weighted) spin-echo pulse sequences. Six microadenomas were demonstrated with MR. Four tumors had a long T1 and long T2, one had a long T1 and short T2, and one bromocriptine-treated tumor had a short T1 and short T2. MR failed to delineate the focal mass within the pituitary gland in the other five patients. The cause of the MR failures was not determined; however, tumor size did not appear to be a factor. MR signals arising from microadenomas are variable and possibly related to the activity of the prolactin-secreting cells. Correlation of findings from MR imaging, spectroscopy, and electron microscopy may lead to an understanding of the variable MR appearance of microadenomas.

Spinal epidural lymphomas: CT features in seven patients.

The computed tomographic (CT) images and records of seven patients with epidural lymphomas were reviewed to analyze their CT characteristics. Although no single specific pattern was identified, several signs were seen: (1) an intraspinal epidural irregular mass extending over several vertebral levels; (2) a mass with osteolysis or sclerosis; and (3) a combined intra- and extraspinal mass. Lymphoma must be considered in epidural masses when there are associated bone changes.

Evidence of a critical leucite particle size for microcracking in dental porcelains.

The leucite particles in dental porcelains are often partially encircled by microcracks that are the result of the thermal expansion mismatch between leucite and the surrounding glass matrix. Although the magnitude of the stress at the particle-matrix interface is independent of the particle size (Selsing, 1961), Davidge and Green (1968) showed experimentally that there is a critical particle size below which microcracking is absent. The critical particle size is explained by a Griffith-type energy balance criterion: Below the critical size, the stress magnitude may be sufficient to cause cracking, but there is insufficient strain energy for the creation of the new surfaces of the microcrack. The purpose of the present study was to determine whether the mean leucite particle size of a dental porcelain influences the degree of microcracking in the porcelain. Microcrack density, leucite particle surface area per unit volume, and leucite mean volume-surface diameter, D3,2, were determined by quantitative stereology on 10 specimens each of 6 dental porcelains and Component No. 1 of the Weinstein et al. patent (US Patent 3,052,982, 1962). The fraction of leucite particles with microcracks around them, f(mc), was estimated for each porcelain from the microcrack density and the leucite surface area. Using the equations of Selsing (1961) and Davidge and Green (1968), we calculated the critical particle diameter, Dc, for leucite to be 4 microm. The porcelains were partitioned according to whether their mean leucite particle diameters, D3,2, fell above or below Dc, and their values of f(mc) were analyzed by a permutation test with random re-sampling. The porcelains with mean leucite particle diameters below Dc had a significantly lower fraction of cracked particles compared with the porcelains with mean leucite particle diameters above Dc (p < 0.05). This study provides evidence that microcracking in dental porcelain can be minimized by a reduction of the mean leucite particle diameter to less than 4 microm.

A comparison of MR sequences for lesions of the parotid gland.

PURPOSE: To compare six MR sequences (plain and gadolinium-enhanced fat suppressed T1-weighted spin echo, T2-weighted standard spin echo, fat-suppressed and non-fat-suppressed T2-weighted fast spin echo, and inversion-recovery T2-weighted fast spin echo) in their ability to detect, delineate, and characterize lesions of the parotid gland. METHODS: Fifty-eight parotid gland lesions imaged on 47 examinations were retrospectively evaluated by three blinded observers. Several outcome-related variables were compared by the above six sequences: imaging time, image quality, anatomic sharpness of parotid space, subjective lesion conspicuity, detected abnormality volume, number of individual lesions or discrete lobulations, conspicuity of invasion into adjacent boundaries and structures, and overall diagnostic value. RESULTS: Differences in the above outcome variables between sequences did not correlate with MR scanner software upgrade level, coil type, or lesion-dependent characteristics. Fat-suppressed fast spin-echo T2-weighted and inversion-recovery fast spin-echo T2-weighted sequences resulted in significantly higher scores for lesion conspicuity, detected abnormality volume, and overall diagnostic value. T1-weighted images resulted in the next highest scores, whereas gadolinium-enhanced T1-weighted and standard spin-echo T2-weighted sequences performed poorly for most parotid lesions. CONCLUSION: MR imaging of the parotid gland should include fat-suppressed, long-repetition-time, fast spin-echo T2-weighted, and T1-weighted sequences. Gadolinium-enhanced images need not be obtained routinely.