Preoperative routine evaluation of bilateral adrenal glands by endoscopic ultrasound and fine-needle aspiration in patients with potentially resectable lung cancer.

BACKGROUND AND STUDY AIMS: The aim of the current study was to assess the detection rate of the right adrenal gland and the diagnostic ability of endoscopic ultrasound (EUS) and fine-needle aspiration (FNA) for the diagnosis of adrenal metastasis in potentially resectable lung cancer. PATIENTS AND METHODS: This retrospective cohort study included a consecutive series of 150 patients undergoing EUS/EUS - FNA for staging of lung cancer. The detection rate of the right adrenal gland by EUS and the diagnostic accuracies of computed tomography (CT), positron emission tomography-CT (PET-CT), and EUS/EUS - FNA for the diagnosis of adrenal metastasis were evaluated. RESULTS: The right adrenal gland was visualized by EUS in 131 patients (87.3 %); the left adrenal gland was visualized in all patients. Findings suggestive of metastasis in either one of the adrenal glands or in both were observed in 6 patients (4.0 %) by CT, in 5 patients (3.3 %) by PET-CT, and in 11 patients (7.3 %) by EUS. EUS - FNA was performed simultaneously in the 11 patients, and in 4 patients the diagnosis of metastasis was established. The accuracy for the diagnosis of adrenal metastasis was 100 % for EUS/EUS - FNA, 96.0 % for CT, and 97.0 % for PET-CT (P = 0.1146). CONCLUSIONS: As well as the left adrenal gland, the right adrenal gland was also usually visible by EUS. EUS/EUS - FNA provided an accurate diagnosis of adrenal metastasis, although the prevalence of adrenal metastasis was relatively low in these patients with potentially resectable lung cancer.

Thermostabilization of a chimeric enzyme by residue substitutions: four amino acid residues in loop regions are responsible for the thermostability of Thermus thermophilus isopropylmalate dehydrogenase.

A chimeric 3-isopropylmalate dehydrogenase, named 2T2M6T, made of parts from an extreme thermophile, Thermus thermophilus, and a mesophile, Bacillus subtilis, was found to be considerably more labile than the T. thermophilus wild-type isopropylmalate dehydrogenase. In order to identify the molecular basis of the thermal stability of the T. thermophilus isopropylmalate dehydrogenase, 11 amino acid residues in the mesophilic portion of the chimera were substituted by the corresponding residues of the T. thermophilus enzyme, and the effects of the side chain substitutions were analyzed by comparing the reaction rate of irreversible heat denaturation and catalytic parameters of the mutant chimeras with those of the original chimera, 2T2M6T. Four single-site mutants were successfully stabilized without any loss of the catalytic function. All these four sites are located in loop regions of the enzyme. Our results strongly suggest the importance of these loop structures to the extreme stability of the T. thermophilus isopropylmalate dehydrogenase.

The transition of cadherin expression in osteoblast differentiation from mesenchymal cells: consistent expression of cadherin-11 in osteoblast lineage.

Osteoblasts are derived originally from pluripotent mesenchymal stem cells on migration into the bone matrix. To elucidate the contribution of classical cadherins in this differentiation pathway, we developed a new protocol for their analysis and studied their specific expressions in various cell lines of the mesenchymal lineage, including osteoblasts. N-cadherin was expressed constitutively in all cell lines examined except an osteocyte-like cell line whereas cadherin-11 was expressed selectively in preosteoblast and preadipocyte cell lines. P-cadherin also was expressed in primary cultures of calvarial cells and mature osteoblasts at a relatively low level compared with N-cadherin and cadherin-11. M-cadherin was expressed only in a premyoblast cell line. We observed the transition of cadherin expression from M-cadherin to cadherin-11 in the premyoblast cell line when osteogenic differentiation was induced by treatment with bone morphogenetic protein 2 (BMP-2), while the expression of N-cadherin remained unchanged. In contrast, when a preadipocyte cell line, which shows a similar pattern of cadherin expression to osteoblasts, was induced to undergo adipogenic differentiation, the expression of N-cadherin and cadherin-11 was decreased. These observations characterize the cadherin expression profile of mesenchymal lineage cells, especially osteoblasts, which regularly express cadherin-11. Cadherin-11 may affect cell sorting, alignment, and separation through differentiation.

Expression and function of the splice variant of the human cadherin-11 gene in subordination to intact cadherin-11.

Cadherin-11, a member of the type II classic cadherin subfamily, differs from type I family molecules such as P-, E-, and N-cadherins. An isoform of the human cadherin-11 gene, termed the variant form, encodes a truncated protein with a different cytoplasmic domain. The resulting protein does not possess any part of the cytoplasmic domain common to other cadherins. In the present study, analysis of the genomic organization of the cadherin-11 gene revealed that an insertion of 179 bp in an intron generates an alternatively spliced form. The mRNA expression of the variant form of cadherin-11 was examined in normal tissues by reverse transcription-polymerase chain reaction and/or Northern blot analyses. The variant form was expressed in the heart, brain, placenta, lung, and bone, but not in the kidney, skeletal muscle, pancreas, and liver. Western blot analyses revealed that the variant form is expressed as an 85 kDa protein, and that an additional secreted form also exists as an 80 kDa protein originated from cleavage of the intact form. Gene transfer of the variant form into L cells demonstrated that it lacked the adhesion properties characteristic of the intact form of cadherin-11 but enhanced the activity of Ca2+-dependent adhesion of the intact form of cadherin-11. The variant was expressed on the surface together with the intact form and stabilized the interaction between the intact form and beta-catenin. These findings suggest that expression of the variant form of human cadherin-11 may regulate the intact cadherin-11-mediated adhesion and alter the morphogenetic processes during mesenchymal cell differentiation including osteoblasts.

Targeted disruption of cadherin-11 leads to a reduction in bone density in calvaria and long bone metaphyses.

The migration and adhesion of osteoblasts requires several classical cadherins. Cadherin-11, one of the classical cadherins, was expressed in mouse osteoblasts in skull bone and femur, revealed by immunohistochemistry. To elucidate the function of cadherin-11 in osteoblastogenesis, cadherin-11 null mutant mice were investigated. Although apparently normal at birth, Alizarin red staining of null mutant mice showed a reduced calcified area at the frontal suture that caused a round-shaped calvaria with increasing animal age to 3 months. Consequently, there was a reduction in bone density at the femoral metaphyses and the diploë of calvaria in null mutant mice. In the in vitro culture of newborn calvarial cells, the calcified area of mutant cells was smaller than those derived from wild-type littermates. These results show that absence of cadherin-11 leads to reduced bone density in some parts of skeletons including calvaria and long bone metaphyses, and thus suggest that cadherin-11 plays roles in the regulation of osteoblast differentiation and in the mineralization of the osteoid matrix.

Large-scale preparation of recombinant human calcitonin from a multimeric fusion protein produced in Escherichia coli.

In order to develop a large-scale, high-yield production process for human calcitonin (hCT) in Escherichia coli, a stable expression plasmid was constructed and the expressed protein was modified for efficient cleavage by protease. Multiple copies of a synthetic gene encoding hCT-Leu-Arg, a substrate for C-terminal amidation by carboxypeptidase Y (CPY), were inserted into the stable expression plasmid. Using this plasmid, the expression of a multimeric fusion protein was induced by shifting the temperature from 34 to 38 degrees C. The multimeric fusion protein was accumulated as inclusion bodies. After the fermentation, the cells were harvested and disrupted by a homogenizer. The insoluble multimeric fusion protein was suspended in 6.6 M urea solution. At this time, however, the protein could not be solubilized and thus the efficiency of cleavage by protease was low. To solubilize the protein and protect Lys residues against digestion by trypsin, the protein was citraconilated with citraconic anhydride. Following S-sulfonation with Na2SO3-CuSO4, almost all the protein was solubilized. The solubilized, citraconilated, and S-sulfonated protein was digested with trypsin efficiently. By treatment with trypsin, the multimeric fusion protein was cleaved into monomers of citraconilated and S-sulfonated hCT-Leu-Arg (S-hCT-Leu-Arg). The subsequent decitraconilation was performed at low pH. S-hCT-Leu-Arg, isolated by preparative HPLC, was directly converted into S-hCT-HN2 by CPY without removal of the Arg residue. Finally, S-hCT-NH2 was desulfonated and converted into mature hCT. In this way, a large amount of recombinant mature hCT was obtained in a tank fermentation. To our knowledge, this is the first report of industrial-scale, human calcitonin production using a multimeric fusion protein expression system.

Prevalence of Listeria (spp.) in wild rats captured in the Kanto area of Japan.

In the Kanto area a total of 245 wild rats were captured. All rats captured in Ikebukuro (110 rats) and 9 out of 41 rats in Yokohama were Rattus rattus, and all other 126 rats were Rattus norvegicus. In Kashima and Ikebukuro, listeria was isolated from 28 rats (77.8%) and 27 rats (24.5%), respectively, but in the other 4 areas listeria was isolated from 0-7% rats. Listeria monocytogenes was isolated from 12 rats (10.9%) captured in Ikebukuro and 2 rats in Kashima and Numazu. The frequent isolation of L. monocytogenes in buildings suggests the possibility of R. rattus as a reservoir of L. monocytogenes and the continual environmental contamination in buildings by L. monocytogenes.

Seroepidemiological survey of lymphocytic choriomeningitis virus in wild house mouse (Mus musculus) in Yokohama Port, Japan.

From 1985 to 1989, a total of 129 mice was captured from 7 piers of Yokohama port. Of these, 9 (7.0%) were positive to lymphocytic choriomeningitis virus (LCMV) antigen in indirect fluorescence antibody test. Six out of 31 mice (19.4%) in 1985 and 3 out of 23 mice (13.0%) captured in 1986 were positive. All the mice (74) captured in 1988 and 1989 were negative. Although 7 out of 17 mice (41.2%) in Osanbashi-Shinko pier and 2 out of 23 (8.7%) in Honmoku pier were positive in 1985 and 1986, all mice captured in other piers were negative. This is the first report detecting LCMV antibody in wild house mice in Japan.

Effect of patellar taping on knee kinetics of patients with patellofemoral pain syndrome.

STUDY DESIGN: Single-group repeated measures examining patients with patellofemoral pain syndrome. OBJECTIVE: To examine the effect of McConnell patellar taping on single-leg vertical jump height and knee extensor moment and power during a vertical jump and lateral step-up. BACKGROUND: Patellar taping is used by clinicians in an attempt to maximize knee extensor function during sporting activities and rehabilitation of patients with patellofemoral pain syndrome. No studies have examined the effect of patellar taping on vertical jump height and knee extensor moment and power during a maximal vertical jump or lateral step-up. METHODS AND MEASURES: Fourteen women (24.4 +/- 5.8 years) with unilateral patellofemoral pain performed a single-leg vertical jump and lateral step-up during 4 knee conditions: patellar tape, placebo tape, no tape, and the uninvolved knee. Maximal knee extensor moment, knee power, and vertical jump height were measured for each condition using a force platform and motion analysis system. RESULTS: Analysis of variance and post hoc analyses revealed a main effect for knee condition. The patellar tape condition resulted in a greater knee extensor moment (1.57 +/- 0.32 N.m/kg) and power (3.47 +/- 0.67 W/kg) than did the no-tape (1.31 +/- 0.39 N.m/kg and 2.79 +/- 1.21 W/kg) and placebo tape (1.33 +/- 0.30 N.m/kg and 2.70 +/- 0.99 W/kg) conditions. Additional analyses showed that the vertical-jump height was significantly greater in the uninvolved lower extremity (25.69 +/- 2.66 cm) compared with the patellar tape (23.33 +/- 4.22 cm), placebo tape (23.08 +/- 4.20 cm), and no-tape (23.45 +/- 4.54 cm) conditions. The patellar tape condition did not show a different vertical jump height than the placebo or no-tape conditions. CONCLUSIONS: These results suggest that patellar taping compared with no tape may improve the knee extensor moment and power during weight-bearing activities such as the lateral step-up exercise and the vertical jump.

[Conscious levels of processing a preceding stimulus and priming effect].

This study was conducted to investigate how two different conditions of processing a prime would affect the lexical decision time of a target. One condition was called the conscious condition, where a prime was presented for 250 ms and a subject was instructed to expect any target from that prime. Another condition was the unconscious condition, where a prime was briefly exposed and followed by a pattern mask so that a subject could not identify it. The effect of an associative strength of prime-target relatedness was also investigated. The associative strength of prime-target relatedness in the related condition was relatively weak in Exp. I, and strong in Exp. II. In the conscious condition, facilitation and inhibition were found both in Exp. I and in Exp. II. On the other hand, in the unconscious condition, only inhibition was found in Exp. I, but only facilitation in Exp. II. These results suggest that automatic activation and attention system were both needed to account for priming effect.

[Semantic priming effect on the perception of a Kanji].

A priming experiment was conducted to investigate how semantic context affects the perception of a briefly exposed Kanji target. On each trial, a Kanji or a plus (neutral) was presented as the prime stimulus for 1 s. When the prime was a Kanji, subjects were required to imagine any compound word, the first letter of which was to be that prime. After a blank of 500 ms, a target Kanji was briefly exposed followed by a patterned mask and two response alternatives, one of which the subject was asked to choose. The duration of the target was so adjusted that each subject could respond correctly at approximately 75% level. The % choice response was higher when the prime and the target made up a compound word (related condition) and also matched to the imagined word (facilitation) than when not (neutral condition). When the prime and the target were unrelated, the performance was the worst (inhibition). In the related condition, however, responses were more accurate even when the prime-target pair did not match to the imagined compound word than in the unrelated condition. These results suggest that semantic priming effect occurs automatically regardless of whether the subject may expect the target or not.

[A temporal feature of memory for plans].

How do we remember future plans? In this study, three experiments were conducted to examine this issue. Thirty to forty different undergraduate or graduate students participated as subjects in each experiment. In Experiment 1, subjects were asked to memorize plans for a day, assuming that they were of tomorrow in the 'future' condition and that they were of yesterday in the 'past' condition. The result showed that plans for the morning and the evening were recalled better than plans for the daytime (the U-shape effect), only in the future condition. In Experiment 2, plans were presented without time, so that subjects could not use any specific schema associated with time. In this condition, the U-shape effect disappeared. In Experiment 3, subjects were required to memorize plans for two days, tomorrow and the day after tomorrow. The result showed the U-shape effect for each day, not for two consecutive days. These results lead to the conclusion that some 'temporal information' about a day may affect the memory for future plans.

[Measurement of the feeling of knowing: norms of 216 general-information questions].

The purpose of this study is to provide the normalized list of questions for investigating the feeling of knowing. Normative data were collected on 216 general-information questions from a wide variety of topics, including history, sports, art, literature, and so on. Three hundred and sixty-three undergraduates made a one-word response to each question, or rated their feeling of knowing on a 7-point scale if they could not recall any responses. The results showed that the questions distributed over a wide range on a scale of difficulty. There were few incorrect responses to the questions. Feeling-of-knowing ratings were moderately correlated with the probabilities of correct answers. These findings suggested that the questions provided in this study would serve for the research on the feeling of knowing. It was also suggested that the use of easier questions should generate a strong feeling of knowing.

[The effects of conscious and subconscious processing of hostility- or friendliness-related words on the personality impression of others].

Four experiments were conducted to investigate how the prior processing of trait relevant information influenced upon the impression formation. Twenty university students participated in each experiment. First subjects performed a cognitive task in which they processed hostility (Exp. I, II) or friendliness (Exp. III, IV) related words, consciously (Exp. I, III) or subconsciously (Exp. II, IV). In another ostensibly unrelated task, subjects rated a stimulus person on several trait scales based on an ambiguous behavioral description, regarding to hostility (Exp. I, II) or friendliness (Exp. III, IV). It was shown that the more hostility words subjects processed either consciously or subconsciously, the more extreme and negative ratings they yielded. As for friendliness words, however, such effects were found only when they were subconsciously processed, not when they were consciously processed. It was argued that conscious processing was affected by positiveness or negativeness of trait words, but subconscious processing was not.

Incidence of clavicular rhomboid fossa (impression for costoclavicular ligament) in the Brazilian population: forensic application.

In the last years, anthropology has been widely explored mainly when related to bones due to its morphologic characteristics, such as the rhomboid fossa of the clavicle. This study examined the incidence of the rhomboid fossa in paired clavicles of Brazilian subjects obtained from 209 adult bodies of known age and sex (107 males and 102 females) on which postmortem examinations had been performed by the senior author. The data were submitted to qualitative statistical analysis according to Fisher. There was a statistical difference (p= 5.98 x 10-23) between sexes related to the frequency of the rhomboid fossa. The fossa was absent in 97,1% of the female clavicles and the incidence of bilateral fossa was present in 2,9% of females. The incidence of bilateral fossa was 29% for male clavicles. The sexual or side differences in the incidence of the fossa could be found in this study, and qualitative analysis can corroborate sex determination of unidentified bodies in forensic medicine.

The rubble-pile asteroid Itokawa as observed by Hayabusa.

During the interval from September through early December 2005, the Hayabusa spacecraft was in close proximity to near-Earth asteroid 25143 Itokawa, and a variety of data were taken on its shape, mass, and surface topography as well as its mineralogic and elemental abundances. The asteroid's orthogonal axes are 535, 294, and 209 meters, the mass is 3.51 x 10(10) kilograms, and the estimated bulk density is 1.9 +/- 0.13 grams per cubic centimeter. The correspondence between the smooth areas on the surface (Muses Sea and Sagamihara) and the gravitationally low regions suggests mass movement and an effective resurfacing process by impact jolting. Itokawa is considered to be a rubble-pile body because of its low bulk density, high porosity, boulder-rich appearance, and shape. The existence of very large boulders and pillars suggests an early collisional breakup of a preexisting parent asteroid followed by a re-agglomeration into a rubble-pile object.

Touchdown of the Hayabusa spacecraft at the Muses Sea on Itokawa.

After global observations of asteroid 25143 Itokawa by the Hayabusa spacecraft, we selected the smooth terrain of the Muses Sea for two touchdowns carried out on 19 and 25 November 2005 UTC for the first asteroid sample collection with an impact sampling mechanism. Here, we report initial findings about geological features, surface condition, regolith grain size, compositional variation, and constraints on the physical properties of this site by using both scientific and housekeeping data during the descent sequence of the first touchdown. Close-up images revealed the first touchdown site as a regolith field densely filled with size-sorted, millimeter- to centimeter-sized grains.

PT pretreatment inhibits 48/80-induced activation of Ca(+)-permeable channels in rat peritoneal mast cells.

We recently reported that the secretagogue, compound 48/80, activated Ca2(+)-permeable channels of mast cells possibly via a second messenger [Kuno, Okada, and Shibata. Am. J. Physiol. 256 (Cell Physiol. 25): C560-C568, 1989]. The effects of pretreatment with pertussis toxin (PT) on compound 48/80 (48/80)-induced activation of the Ca2(+)-permeable channel have now been investigated by measuring Ca2+ signals of cell suspensions using the Ca2+ indicator fura-2 and by recording Ba2+ currents through the channel using the patch-clamp technique. In the presence of extracellular Ca2+, the fluorescence change was biphasic, with an immediate rise and a delayed peak at room temperature. The delayed peak, mainly due to Ca2+ entry through plasma membranes, was greatly reduced by pretreatment with PT. The quenching of the fluorescence by 48/80-induced Mn2+ influx was also decreased by PT, whereas the Ca2+ transients due to Ca2+ release from the intracellular stores apparently did not change. In patch-clamp recordings from cell-attached patches with pipettes containing isotonic Ba2+, the 48/80-induced Ba2+ currents were either suppressed or delayed in the PT-treated cells, under conditions where degranulation was absent. These results suggest that PT-sensitive GTP-binding protein is involved in activating the Ca2(+)-permeable channel in mast cells during stimulus-secretion coupling.