Influence of double layer filter on mean glandular dose (MGD) and image quality in low energy image of contrast enhanced spectral mammography (LE-CESM).

INTRODUCTION: The aim of this study was to evaluate mean glandular dose (MGD) and image quality in low energy imaging from contrast-enhanced spectral mammography (CESM) when using double-layer filtration. METHODOLOGY: A dedicated phantom was used to quantitatively estimate the MGD and image quality. The target slab of the phantom consisted of three iodine coins having a concentration of 1.0 mgI/cm3, 2.0 mgI/cm3, 4.0 mgI/cm3, a 100% adipose equivalent coin and a 100% glandular equivalent coin. The phantom was exposed using a semiautomated function at 28 k, 30 kV and 32 kV. MGD. Contrast to noise ratio (CNR) and figure of merit (FOM) were estimated for Mo/Rh, Mo/Rh + Cu, Mo/Rh + Al and Mo/Rh + Cd combinations using three breast equivalent compositions. RESULTS: MGD was reduced up to a maximum of 1.03 mGy from 1.17 mGy for 100% adipose tissue. 1.18 mGy from 1.34 mGy for 50% glandular tissue and 1.39 mGy from 1.72 mGy for the 100% glandular phantom when using double-layer filtration. All of the above-mentioned results were obtained for the 50 mm phantom using 32 kV. CNR and FOM values were not significantly reduced with a double-layer filter when compared to a single-layer filter. CONCLUSION: The present study concluded that Mo/Rh + Cu is the best combination to reduce the MGD significantly when compared to Mo/Rh + Al or Mo/Rh + Cd. Mo/Rh + Cu also achieved optimal image quality when compared to the Mo/Rh single filter combination. IMPLICATIONS OF PRACTICE: The use of a double-layer filter in low energy imaging of CESM results in a significant reduction in MGD without degrading the quality of the image.

Enhanced inhibitory effects of TBT chloride on the development of F1 rats.

Neurotoxicity is one of the major effects of tributyltin (TBT). The effects on the next generation of F(1) rats exposed to TBT via the placenta and their dams' milk may be stronger than those on adults. Pregnant Wister rats were exposed to TBT at 0 and 125 ppm in their food. Half of the female F(1) rats in both groups were exposed to TBT at 125 ppm in their food from 9 to 15 weeks of age. Female F(1) rats were divided into the following groups: the control-control (CC) group, with no exposure; the TBT-control (TC) group, exposed to TBT via the placenta and their dams' milk; the control-TBT (CT) group, exposed to TBT via their food from 9 to 15 weeks of age; and the TBT-TBT (TT) group, exposed to TBT via the placenta, their dams' milk, and their food (n = 10/group). After administration, an open-field test and prepulse inhibition (PPI) test were performed at 15 weeks of age. The mean body weights of the TC and TT groups were significantly lower than that of the CC group from 9 to 15 weeks of age. The mean relative thymus weight of the TC and TT groups was significantly lower than that of the CC group. In the open-field test, a marked decrease in the total locomotion distance was observed in the TT group. The mean values in the TT and TC groups were significantly lower than that in the CC group. For the locomotion distance between 15 and 20 min, the mean values in the CT, TC, and TT groups were significantly lower than that in the CC group. The mean locomotor distance between 25 and 30 min in the TT group was significantly lower than that in the CC and TC groups. The mean values of instances of wall rearing in the TC, CT, and TT groups were significantly lower than that in the CC group. The mean value of face washing or body washing in the TT group was significantly lower than that in the CT group. There were no significant differences in indexes of the PPI test. Exposure to TBT via the placenta and their dams' milk inhibited the development of F(1) rats, which continued after weaning. Inhibition of the rats' activity induced by exposure to TBT via the placenta and their dams' milk and/or via their food was suggested. The effects were most evident in the TT group.

Solution structure of the activator contact domain of the RNA polymerase alpha subunit.

The structure of the carboxyl-terminal domain of the Escherichia coli RNA polymerase alpha subunit (alpha CTD), which is regarded as the contact site for transcription activator proteins and for the promoter UP element, was determined by nuclear magnetic resonance spectroscopy. Its compact structure of four helices and two long arms enclosing its hydrophobic core shows a folding topology distinct from those of other DNA-binding proteins. The UP element binding site was found on the surface comprising helix 1, the amino-terminal end of helix 4, and the preceding loop. Mutation experiments indicated that the contact sites for transcription activator proteins are also on the same surface.

Thymic hyperplasia in patients with Graves' disease. Identification of thyrotropin receptors in human thymus.

Thymic size and density were studied in 23 untreated patients with Graves' disease and 38 control subjects using computed tomography. Both thymic size and density were higher in untreated patients with Graves' disease than in control subjects in the age-matched group. After treatment with antithyroid drugs, both thymic size and density were significantly reduced, with a concomitant decrease in thyrotropin receptor antibodies. PCR of human thymic cDNA using primers for human thyrotropin receptor amplified a fragment in a size expected for the receptor, and its nucleotide sequence was identical to human thyrotropin receptor cDNA in the thyroid. Northern blot analysis of human thymic poly(A)+ RNA demonstrated the presence of the full length form of thyrotropin receptor mRNA. Western blot analysis of human thymic membrane using anti-thyrotropin receptor peptide antibodies demonstrated a band of 100 kD that was also observed in the thyroid membrane. Immunohistochemistry of thymic tissue using mouse antihuman thyrotropin receptor monoclonal antibodies demonstrated the immunostaining of epithelial cells. These results indicate that thymic hyperplasia is apparently associated with Graves' disease and suggest that thymic thyrotropin receptor may act as an autoantigen that may be involved in the pathophysiology of development of Graves' disease.

Structural map of the alpha subunit of Escherichia coli RNA polymerase: structural domains identified by proteolytic cleavage.

The alpha subunit of Escherichia coli RNA polymerase plays essential roles in protein-protein contacts, not only for RNA polymerase assembly, but also for transcription activation by class I factors. To reveal the structure-function relationship of the alpha subunit, we attempted to elucidate the organization of the structural domains by analysis of the pattern of limited proteolysis with two endoproteases, V8 protease and trypsin. The results indicate that one region, Arg235 to Glu244, is highly accessible to endoproteases. We propose that the alpha subunit consists of two major structural domains, the amino-terminal domain upstream from Arg235 and the carboxy-terminal domain downstream from Glu245, each being connected by an inter-domain linker formed by the spacer between these two amino acid residues. The structural organization is in good agreement with its functional map, i.e., the amino-terminal subunit assembly determinants and the carboxy-terminal transcription activation determinants, including the contact sites with class I transcription factors and DNA UP (enhancer) elements. The secondary proteolytic cleavage sites were also determined, in order to analyse intra-domain structures.

Localization of ubiquitin carboxyl-terminal hydrolase-L1 in cynomolgus monkey placentas.

Ubiquitin carboxyl-terminal hydrolase-L1 (UCH-L1) is a restrictedly expressed enzyme in neural and reproductive tissues, and it is considered to have a significant role in reproduction. In the present study, we investigated the localization of UCH-L1 in placenta of cynomolgus monkeys (Macaca fascicularis). UCH-L1 protein was detected in cytotrophoblasts of chorionic plate and villi, and decidual cells of decidua basalis in cynomolgus monkey placenta, and the amount of UCH-L1 protein in whole placenta increased as pregnancy progressed. These results supported that UCH-L1 is necessary for placental and fetal development in primate placenta. This is the first report to demonstrate the presence of UCH-L1 in primate placenta, and the cynomolgus monkey may be a useful model for the study of the functions of the ubiquitin-proteasome system in human pregnancy.

Ceramide enhances growth hormone (GH)-releasing hormone-stimulated cyclic adenosine 3',5'-monophosphate accumulation but inhibits GH release in rat anterior pituitary cells.

In this study, the effect of ceramide on GH-releasing hormone (GHRH)-stimulated cAMP accumulation and GH release in rat anterior pituitary cells was investigated. C2-, C6-, and C8-ceramide were found to enhance GHRH-stimulated cAMP accumulation. In contrast, their effects on GHRH-stimulated GH release were inhibitory. Treatment with a glucosylceramide synthase inhibitor produced a similar enhancing effect on cAMP accumulation and an inhibitory effect on GH release. To identify the pathway through which ceramide mediated its effect, it was found that ceramide inhibited GH release stimulated by KCl, BayK 8644, and a GH-releasing peptide, but not that stimulated by ionomycin or an activator of protein kinase C. Direct measurement of intracellular Ca2+ revealed that C2-ceramide inhibited GHRH- and KCl-mediated increases in intracellular Ca2+, suggesting that ceramide probably inhibits GH release through inhibition of the L-type Ca2+ channels. As for its mechanism on cAMP accumulation, the enhancing effect of ceramide on GHRH-stimulated cAMP accumulation was abolished in the presence of a phosphodiesterase inhibitor, isobutylmethylxanthine, suggesting that ceramide enhances the cAMP response through inhibition of its metabolism. Taken together, our results suggest that ceramide plays an important role in the regulation of GHRH-stimulated responses in somatotrophs. By reducing GH secretion while enhancing cAMP accumulation, ceramide may promote the synthesis and storage of GH in rat anterior pituitary cells.

Ceramide inhibits L-type calcium channel currents in rat pinealocytes.

In rat pinealocytes, ceramide can inhibit the KCl- and BayK 8644-mediated potentiation of cAMP and cGMP accumulation, suggesting that the L-type Ca2+ channel is a target of ceramide action. This was examined in the present study using intracellular Ca2+ measurement and patch-clamp studies. In fura-2-loaded pinealocytes, C2- and C6-ceramide inhibited the Ca2+ increase caused by BayK 8644 and KCl, but not that caused by norepinephrine, suggesting an inhibitory effect of ceramide on the L-type Ca2+ channels. Patch-clamp analysis confirmed that C2- and C6-ceramide, but not C2-dihydroceramide (the inactive analog) inhibited the L-type Ca2+ channel current. Furthermore, treatments known to increase cellular ceramide levels, including a glucosylceramide synthase inhibitor and sphingomyelinase, also inhibited this current. The inhibitory effect of ceramide on the current was attenuated by lavendustin A, a tyrosine kinase inhibitor, but not by H7, a serine/threonine kinase inhibitor. The effect of ceramide was mimicked by interleukin-1beta, a cytokine highly expressed in the pineal that is known to activate the sphingomyelin pathway. These results indicate that the sphingomyelin pathway is another important signaling mechanism that regulates the L-type Ca2+ channel, and tyrosine kinase appears to be involved in the effect of ceramide.

Activated T lymphocyte subsets in experimental allergic neuritis.

Changes in activated T cell subsets in peripheral blood were examined during the course of experimental allergic neuritis (EAN), using two-color immunofluorescence flow cytometry. Both CD4+ and CD8+ activated T cells decreased transiently before the onset of clinical signs, and increased just around the time of onset of the disease. In contrast, during the recovery phase, the numbers of CD4+ activated T cells returned to the normal range, whereas CD8+ activated T cells continued to increase. These findings imply that activation of CD4+ helper/inducer cells contributes mainly to the evolution of EAN, and that of CD8+ suppressor cells are necessary for recovery.

Cell-mediated immunity to bovine P2 protein and neuritogenic synthetic peptide in experimental allergic neuritis.

Cellular reactivity to bovine P2 protein (P2) and its two synthetic peptides, SP66-78 and SP70-78, was serially examined by the lymphocyte proliferation test in animals with experimental allergic neuritis (EAN). SP66-78 and SP70-78 correspond to residues 66-78 and 70-78 of bovine P2. Proliferative response to SP66-78 as well as P2 appeared at day 7 before the onset of EAN and was clearly manifested at day 14 in the active stage, thereafter disappearing in the stable stage, whereas no response to SP70-78 was detected during the course of the disease. These results suggest that cell-mediated immune response to P2 and the specific part residues 66-78 of P2 play an important role in the pathogenesis of EAN.

Inhibitory effect of hemin on the mutagenic activities of carcinogens.

An inhibitory effect of hemin on mutagenicities of a range of carcinogens was found by adding hemin to the preincubation mixture of the Ames' test. Strong inhibitions were observed for benzo[alpha]pyrene, 3-methylcholanthrene, 7,10-dimethylbenz[alpha]anthracene, chrysene, 2-acetylaminofluorene, 2-nitrofluorene and aflatoxin B1. Generally, 50% inhibition was caused by an amount of hemin 1--2 equivalents to the mutagen. Excess of hemin caused complete inhibitions. Hemin did not affect the mutagenicities of 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, 4-nitroquinoline-1-oxide, nitromin, N-methyl-N-nitrosourea, N-methyl-N'-nitro-N-nitrosoguanidine, N-nitrosodi-n-butyl-amine, quinoxaline-1,4-dioxide and carbadox. Biliverdin, bilirubin and chlorophyllin were also effective as inhibitors for the mutagenicity of benzo[alpha]pyrene.

Porphyrins as possible preventers of heterocyclic amine carcinogenesis.

Our studies have shown that hemin and chlorophyllin can directly interact with heterocyclic amines (HAs) and prevent their mutagenic actions. Hemin and chlorophyllin can trap HAs efficiently, probably by forming face-to-face complexes with them. The trapping was most clearly demonstrated by use of solid-supported porphyrins, hemin-agarose and chlorophyllin-chitosan. Furthermore, spectroscopic measurements have suggested that there are interactions in solution between the porphyrins and the HAs. A number of in vivo data have been accumulated by efforts from many laboratories for the anticarcinogenic and antigenotoxic properties of porphyrins, particularly chlorophyllin, against HAs.

Inhibitory activity of chlorophyllin on the genotoxicity of carcinogens in Drosophila.

Antimutagenic activity of copper chlorophyllin against various carcinogenic mutagens was assayed with Drosophila genotoxicity tests, i.e., the wing spot test for detecting somatic cell mutations and the DNA repair test for detecting DNA damage. In these tests, Drosophila larvae were fed carcinogens together with chlorophyllin. Polycyclic aromatic compounds, including heterocyclic amines, polycyclic aromatic hydrocarbons, aromatic amines and aromatic nitro compounds, were subject to inhibition, with a few exceptions. The results support the view that chlorophyllin traps carcinogens by forming complexes, thereby inhibiting the absorption of these compounds from the digestive tract. Consistent with this mechanism, Sepharose-supported chlorophyllin in the feed inhibited the Trp-P-2-induced wing spot formation, while Sepharose itself was ineffective.

Enhancement of non-specific resistance to viral infection by muramyldipeptide and its analogs.

Antiviral activity of muramyldipeptide (MDP) and its lipophilic derivatives, B30-MDP and MDP-Lys(L18), was investigated in mice infected with vaccinia virus (VV) and herpes simplex virus type 2 (HSV-2). Mice administered these compounds subcutaneously or orally were protected against VV in tail lesion tests and against HSV-2 in skin lesion tests, respectively. Since in vitro antiviral activity was not demonstrated with these compounds in cultured mammalian cells infected with either VV or HSV-2, host-mediated defense mechanisms may play a role in the activity of the compounds. As for serum interferon (IFN) induction, MDP and its analogs showed no activity in mice, suggesting that IFN does not participate in the antiviral mechanisms against VV and HSV-2. An extrinsic antiviral activity was demonstrated when peritoneal macrophages from the mice administered these compounds were cocultivated with VV-infected 3T3 cells. The results indicate that macrophage activation by MDP and its analogs plays a role in the defense mechanisms against viral infection. This activity was not virus-specific. We also demonstrate that the introduction of lipophilic residue(s) into MDP enhances the antiviral activity of mice against VV and HSV-2.

Isolation and characterization of the human chromosomal gene for prostacyclin-stimulating factor.

Prostacyclin-stimulating factor (PSF) is a protein which acts on vascular endothelial cells and stimulates the production of prostacyclin. Recently, we were able to purify PSF from the conditioned medium of cultured human diploid fibroblasts and clone PSF cDNA. In this study, we screened a human genomic library and isolated genomic clones to determine the structure of the human chromosomal PSF gene. By determining the nucleotide sequence and transcription initiation site of this gene, we found that it comprises 5 exons and 4 introns. Southern hybridization analysis indicated the presence of a single copy of the PSF gene per haploid set of chromosomes. The 300 bp upstream of the transcription initiation site had a very high GC content, and 7 binding sites for the transcription regulating factor Sp1 were present.

Monoclonal antibodies specific to the integral membrane protein P0 of bovine peripheral nerve myelin.

Two monoclonal antibodies (mAbs), 58A and 46E, were generated against the major protein P0 of bovine peripheral nervous system myelin (PNSM). The reactivities of the mAbs were assessed by enzyme-linked immunosorbent assay (ELISA), Western blot, and immunohistochemistry. Both mAbs, 58A and 46E, reacted to PNSM of bovine, human, rat and rabbit, but not to chicken PNSM or the brains of rat and rabbit. In the Western blot, these mAbs showed specific binding to bovine P0 as well as deglycosylated P0, but not to myelin-associated glycoprotein (MAG) of bovine spinal cord. The analyses of the lysylendopeptidase-digested peptides of bovine P0 revealed that the epitopes for the mAbs 58A and 46E were located on the amino acid residues 68-79 and 210-216, respectively. Since the mAbs 58A and 46E recognize the extracellular domain and the cytoplasmic domain of P0, respectively, they could be useful for studies on P0's role in myelin formation, its adhesive properties, and functions of the N-terminal extracellular and C-terminal cytoplasmic domains of the protein.

Parameatal urethral cysts.

Two cases of parameatal urethral cysts in boys are reported. Both had been present from early infancy. The etiology of parameatal urethral cysts is discussed.

Expression and nocturnal increase of type II iodothyronine deiodinase mRNA in rat pineal gland.

It has been demonstrated that thyroxine deiodinating activity is present in rat pineal gland, and its activity increases significantly during the night time. We have studied whether mRNA for type II iodothyronine deiodinase is expressed in rat pineal gland and whether the nocturnal rise of pineal T4 deiodinating activity is due to the change in type II iodothyronine deiodinase mRNA level. Reverse transcription-polymerase chain reaction amplification and Northern blot analyses have demonstrated that type II iodothyronine deiodinase mRNA is expressed in rat pineal gland and its mRNA level increases markedly at midnight. These results suggest that the nocturnal rise in pineal T4 deiodinating activity is due to the change in type II iodothyronine deiodinase mRNA level.

Studies on Ca2+-stimulated adenosine triphosphatase in rat submandibular glands: its distribution and properties.

Intracellular distribution and some characteristics of a Ca2+-stimulated adenosine triphosphatase (ATPase) in rat submandibular glands were investigated. This enzyme was activated by calcium alone, and magnesium was not necessary for its activation. Mg2+-stimulated ATPase also was investigated in the same enzyme preparations.

Early tensile bond strengths of several enamel and dentin bonding systems.

Tensile bond strength tests are commonly used for the evaluation of adhesive dental materials. The majority of these tests are carried out after 24 h of storage in water. However, determination of the early tensile bond strength could be more important, especially in relation to gap formation between the cavity surface and the restorative material. This study investigated the tensile bond strengths of five enamel/dentin bonding systems and two experimental dentin bonding systems. Tensile bond strengths were obtained at one min, ten min, and 24 h after the resin composite was cured. Bond strengths at the early stages were always somewhat less than the 24-hour test results. For the enamel/dentin bonding systems, a significant difference was found between the enamel and dentin bond strengths at all time periods, except with Superbond D-liner and Liner Bond. The experimental group with glyceryl methacrylate as the primer produced a good 24-hour result (14.3 MPa), but the early bond strengths were no different from those in the non-primer-treated groups. It was concluded that this material may actually retard the polymerization of the bonding resin. Previous workers have suggested that a tensile bond strength in the order of 20 MPa is necessary for gap-free restorations to be obtained. Should this be the case, then all of the materials tested, from the aspect of early bond strength, lack the strength for prevention of gap formation, although Superbond D-liner and Liner Bond approached this hypothetical figure. These systems, Superbond D-liner and Liner Bond, also exhibit small differences between the enamel and dentin tensile bond strengths.