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Background In heart failure with reduced ejection fraction (HFrEF), drugs including angiotensin-converting enzyme inhibitors (ACEi)/ angiotensin receptor blockers (ARB), beta-blockers, and mineralocorticoid receptor antagonists (MRA) have been shown to have robust survival benefits. However, these guideline-recommended therapies remain underutilized in clinical practice. Frailty is common in HFrEF and may lead to underprescription of life-saving therapy. Purpose We aimed to investigate the association between physical frailty and the use of evidence-based pharmacological therapy for HFrEF, and the impact of this on prognosis Methods The FLAGSHIP study included patients hospitalized for acute HF and data on physical frailty were collected prospectively. Of the total 3,272 patients registered in the FLAGSHIP study, 1,041 HFrEF patients (70 years; 73% male) with left ventricular ejection fraction ≤40% were analyzed and were divided into 4 groups by severity of frailty: category I (n=371) [least frail], II (n=275), III (n=224), and IV (n=171) [most frail]. Results An ACEi/ARB was prescribed in 76% of category I and 53% of category IV patients; for a beta-blocker these proportions were 94% and 76%, respectively; for an MRA they were 55% and 46%, respectively. The proportion of patients using receiving all 3 drugs decreased as frailty increased, with approximately twice the rate of use of triple therapy in category I patients (40.2%) compared to category IV patients (23.4%) [p Conclusions Prescription of guideline-recommended therapy decreased as the severity of frailty increased in patients with HFrEF. Sub-optimal medical therapy was associated with a worse outcome and underprescription of guideline-recommended therapy may contribute to the poor prognosis associated with frailty. An effective strategy is needed to improve the medical treatment of frail patients with HFrEF. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): This study issupported by a Grant-in-Aid for Scientifi c Research (A) from the Japan Society for the Promotion of Science (16H01862). ToruKondo receives grants from the Uehara Memorial Foundation and the Japanese Heart Failure Society Tsuchiya Foundation forthe research activities at the University of Glasgow.

Background In clinical practice, we often encounter the patients with sarcoidosis showing relatively high in the normal range of serum angiotensin-converting enzyme (ACE) value. Purpose We aimed to examine the serum ACE value of the patients with sarcoidosis and determine the new cut-off value for detecting the patients with sarcoidosis. Methods and results We retrospectively examined all 3781 subjects (51.1% men, 60.1±17.0 y.o.) in whom ACE was measured for any reasons including suspected sarcoidosis between 2009 and 2020 in our hospital. Of 293 patients with sarcoidosis, 101, 212, 84, and 88 were diagnosed as sarcoidosis in heart, lung, skin, and eyes, respectively. After excluding 477 patients taking ACE inhibitor and/or immunosuppression agent or those with any diseases affecting serum ACE levels, we analyzed 3304 subjects including 215 with sarcoidosis. Serum ACEs were 19.6 IU/L [IQR, 15.1–31.5] in the subjects with sarcoidosis and 10.7 [8.4–16.5] in those without (P Conclusion(s) In the current cut-off value of serum ACE, sensitivity for detecting sarcoidosis was comparatively low, though positive predictive value was low when the new-cut-off value was used in our study. Further examinations may be needed for the patients suspected sarcoidosis with relatively high ACE in the normal range. Funding Acknowledgement Type of funding sources: None.

Publisher Summary The chapter discusses the internationalization of Japanese commercial banking and the yen. The internationalization of finance has three aspects. First, it involves the internationalization of domestic currency, that is, a domestic currency also becomes an international vehicle of currency and a reserve currency. Second, it involves the internationalization of banks and other financial institutions, that is, they make transactions in both domestic and foreign currencies with nonresidents and residents. Third, it involves the internationalization of money and capital markets, that is, both short-term and long-term funds move freely into and out of the markets across the borders of the countries and interest rates are determined by market forces. There are two main courses in the internationalization of banking: 1) banks may internationalize themselves in accordance with internationalization of their domestic business customers or 2) banks may initiate internationalization on their own. There are two types of limitations in the process of internationalization of banking—internal and external. Internal limitations are basically management problems, while external limitations arise from general economic conditions and various regulations imposed on banks. According to the survey, the main internal limitations as perceived by the banks are, in order of importance, the shortage of competent staff to carry out internationalization, the inability to collect adequate information and know-how, the lack of an adequate system of operations for internationalized banking business, and the cost of establishing foreign bases such as branches, representative offices, and local banks.

Post-pulmonary embolism (PE) syndrome is an important clinical condition that can affect the long-term prognosis after acute PE.We aimed to evaluate the prevalence of residual pulmonary thrombi and the thrombotic burden 1 year after acute PE, by using our refined computed tomography (CT) imaging method.In this prospective study, patients diagnosed with acute PE were recruited and examinations were conducted at 1 month, 6 months, and 1 year. Especially at 1 year, patients were evaluated multifacetedly, including by laboratory tests, quality-of-life, 6-min walking test, and enhanced CT.Fifty-two patients were enrolled. Two patients (3.8%) developed chronic thromboembolic pulmonary hypertension. A total of 43 patients completed evaluation at 1 year, among whom (74%) had residual thrombi, with a median modified CT obstruction index (mCTOI) of 10.7%. In multivariate analysis, residual thrombi at 1 month was the only factor significantly related to residual thrombi at 1 year (odds ratio, 103.4; 95% confidence interval, 4.2-2542.1). The tricuspid regurgitation pressure gradient ≥60 mmHg and left ventricular end-diastolic dimension at diagnosis were significantly related to mCTOI at 1 year (β = 0.367, P = .003; and β = -0.435, P = .001, respectively).Using our improved CT imaging protocol, we found a high prevalence of residual thrombi 1 year after acute PE. Furthermore, right ventricular overload was related to the thrombotic burden. The long-term treatment strategy of acute PE could be modified to include precise CT imaging.

Background Blood pressure (BP) variability has reportedly been a risk factor for various clinical events. To clarify the influence of BP visit‐to‐visit variability on adverse events in patients with nonvalvular atrial fibrillation, a post hoc analysis of the J‐RHYTHM Registry was performed. Methods and Results Of 7406 outpatients with nonvalvular atrial fibrillation from 158 institutions, 7226 (age, 69.7±9.9 years; men, 70.7%), in whom BP was measured 4 times or more (14.6±5.0 times) during the 2‐year follow‐up period or until occurrence of an event, constituted the study group. SD and coefficient of variation of BP values were calculated as BP variability. Thromboembolism, major hemorrhage, and all‐cause death occurred in 110 (1.5%), 121 (1.7%), and 168 (2.3%) patients, respectively. When patients were divided into quartiles of systolic BP‐SD (P =0.015 for thromboembolism; HR, 2.60, 95% CI, 1.36–4.97, P =0.004 for major hemorrhage; and HR, 1.85, 95% CI, 1.11–3.07, P =0.018 for all‐cause death) after adjusting for components of the CHA 2 DS 2 ‐VASc score, warfarin and antiplatelet use, atrial fibrillation type, BP measurement times, and others. These findings were consistent when BP‐coefficient of variation was used instead of BP‐SD. Conclusions Systolic BP visit‐to‐visit variability was significantly associated with all adverse events in patients with nonvalvular atrial fibrillation. Further studies are needed to clarify the causality between BP variability and adverse outcomes in patients with nonvalvular atrial fibrillation. Registration URL: https://www.umin.ac.jp/ctr/ ; Unique Identifier: UMIN000001569.

Honeycomb-like porous chitosan (CS) films are attractive tools for developing functional materials for filters, catalyses, adsorbents, biomaterials

Background To clarify the influence of hypertension and blood pressure ( BP ) control on thromboembolism and major hemorrhage in patients with nonvalvular atrial fibrillation, a post hoc analysis of the J‐ RHYTHM Registry was performed. Methods and Results A consecutive series of outpatients with atrial fibrillation was enrolled from 158 institutions. Of 7937 patients, 7406 with nonvalvular atrial fibrillation (70.8% men, 69.8±10.0 years) were followed for 2 years or until an event occurred. Hypertension was defined as a systolic BP ≥140 mm Hg, a diastolic BP ≥90 mm Hg, a history of hypertension, and/or antihypertensive drug use. Hypertension was an independent risk factor for major hemorrhage (hazard ratio 1.52, 95% CI 1.05–2.21, P =0.027) but not for thromboembolism (hazard ratio 1.05, 95% CI 0.73–1.52, P =0.787). When patients were divided into quartiles according to their systolic BP at the time closest to the event or at the end of follow‐up (Q1, CI 1.75–4.74, P CI 1.02–2.53, P =0.041) after adjustment for components of CHA 2 DS 2 ‐ VAS c score, warfarin use, and antiplatelet use. A systolic BP of ≥136 mm Hg was an independent risk factor for thromboembolism and major hemorrhage. Conclusions BP control appears to be more important than a history of hypertension and baseline BP values at preventing thromboembolism and major hemorrhage in patients with nonvalvular atrial fibrillation. Clinical Trial Registration URL : http://www.umin.ac.jp/ctr . Unique identifier: UMIN 000001569.

Microwave spectra of six isotopic species of (E)- and (Z)-acetaldehyde oxime (CH3CD NOH, CH3CH 15NOH, CH313CH NOH, 13CH313CH NOH, CD3CH NOH, and CD3CD NOH) were observed in the frequency range of 26.0–40.0 GHz. Adding to normal species and CH3CH NOD, the rs coordinates of H1, N, C1, C2, and H3 atoms (refer to Fig. 1 ) of (E) and (Z) isomers in A-state were calculated using Kraitchman's equation. The structural parameters, three bond lengths and three bond angles of (E)- and (Z)-acetaldehyde oxime in A-state were determined. The structural parameters obtained are as follows: for E isomer, r(C1 N): 1.265(15) A, r(C1–C2): 1.510(11) A, r(C1–H1): 1.110(9) A, ∠C2C1N: 117.7(8)°, ∠H1C1N: 118.1(5)°, and ∠C2C1H1: 124.2(7)°, and for Z isomer, r(C1 N): 1.275(4) A, r(C1–C2): 1.510(14) A, r(C1–H1): 1.091(8) A, ∠C2C1N: 124.3(7)°, ∠H1C1N: 113.6(5)°, and ∠C2C1H1: 122.1(7)°. The bond angle ∠C2C1N in Z isomer is much larger than that of (E) isomer, because of the steric repulsion between the methyl and hydroxyl groups in (Z) isomer. The steric repulsion may be related to the low barrier height (V3) of the methyl group in Z isomer.

近年,鳥取県内においてニホンナシ‘ゴールド二十世紀’などにカビ臭と赤アザ症状を伴う汚れ果症状が多発した.この「カビ梨症」と呼ばれる汚れ果症状の果実から酵母様コロニーを示す2菌が高率に分離された.それらは形態学的観察,生化学的性状調査,26S rDNAのD1/D2領域および5.8S rDNAを含むITS領域の塩基配列解析などの結果から,それぞれAcaromyces ingoldiiおよびMeira sp.であると同定された.これらの担子菌系酵母様菌を健全な‘ゴールド二十世紀’ナシ幼果に噴霧接種した結果,自然発病に類似した症状が再現され,発病果実からは接種菌が再分離された.ナシ汚果病はAlternaria sp.など数種の糸状菌によって引き起こされることが報告されているが,今回の結果から,A. ingoldiiおよびMeira sp.をナシ汚果病の病原として追加する.また,最近の分類体系に従い,ナシ汚果病の病原として記録されているHyalodendron sp.をA. ingoldiiと同一種として取り扱った.

The measurement technique on the pattern placement accuracy of an x-ray mask for synchrotron radiation (SR) lithography has been basically studied using a highly-accurate optical measurement machine (Laser XY-3i). The repeatability and reproducibility obtained as an aim were 15 nm and 18 nm in 3 σ respectively, thus providing sufficient measurement accuracies for x-ray masks to attain a ≤ 60 nm overlay accuracy. It was found that the mask setting direction toward a Laser XY-3i mask holder affected measurement accuracies. It is considered that an x-ray mask holding mechanism plays a main role in mask pattern placement accuracy measurement.

Objective: The value of surgical intervention in the management of hypophysitis remains con- troversial. Patent: We describe a 57-year-old man pre- senting with general fatigue persisting for three months. Endocrine examination revealed hypo- pituitarism and diabetes insipidus. Magnetic reso- nance (MR) imaging showed a dumbbell-shaped pituitary mass lesion without a cystic compo- nent. We partially removed the lesion via a trans- sphenoidal approach. Histological examination yielded the diagnosis of granulomatous hypo- physitis associated with Rathke ' s cleft cyst. No deterioration of pituitary function was observed postoperatively. Twenty months after the sur- gery, the lesion has spontaneously regressed on MR imaging, and he is doing well with continu- ing replacement therapy of hydrocortisone, levo- thyroxine and desmopressin acetate. Conclusion: The diagnosis of hypophysitis, apart from typical lymphocytic hypophysitis, is diffi cult even with a surgical biopsy. Because a small specimen may lead to a diagnosis of non- specifi c hypophysitis, partial removal of the lesion is less invasive and recommended for preservation of the pituitary function.

1.8± 1.3 cm at P2. In the condition UCon, the COG of all subjects displaced to dominant side at both postures. The hip joint angular velocity of Con was significantly higher than that of UCon in both postures (p< 0.01). Moreover, the jerk cost on an anteroposterior direction and the right and left direction was significantly larger Con than UCon in both postures (p< 0.01). The jerk cost on vertical direction was not significantly difference between Con and UCon in both postures. Conclusion(s): The energy cost influenced the formation of COG trajectory during sit-to-stand motion with asymmetrical foot placement. Implications: Rising from chair in asymmetrical foot placement is useful treatment for involved lower extremity of stroke patients. The results of this study gave the evidence of the treatment.

Brazzein is a thermostable, sweet-tasting protein isolated from the fruit of the African plant, Pentadiplandra brazzeana Baillon [1]. Brazzein consists of 54 amino acid residues and has four intramolecular disulfide bonds [2]. It is wellknown that the carboxyl groups of sweet compounds participate in binding with the receptor. Brazzein contains five Asp residues at positions 2, 25, 29, 40 and 50, and four Glu residues at positions 9, 36, 41 and 53. The roles of these acidic amino acids were investigated by replacing each with an Ala residue by the fluoren-9-yl-methoxycarbonyl (Fmoc) solid-phase synthesis.

In this paper, a flexible robotic assembly system with decentralized architecture is presented. The system is designed to achieve high reconfigurability so that it can adapt to changes in manufacturing environment; a new robot can be easily installed to the system and execute assembly tasks immediately, together with other devices. For easier reconfiguration, a semi-automated calibration method for positions of newly installed robots is integrated into the system. Our implementation of the assembly system consists of conventional manipulators and a belt conveyor. In the experiment, a new mobile manipulator was installed successfully and performed an assembly operation jointly with other existing manipulators.

In this paper, a robotic assembly system with decentralized architecture is studied. In order to achieve high flexibility and reconfigurability, the system consists of autonomous agents, "holons"; each of assembly devices is controlled by its own holon, "execution holon" and each of assembly operations is handled by a holon, "operation holon". Operation holons are dynamically created and destroyed to execute assembly tasks. When an assembly task is launched in the system, the task is decomposed recursively into primitive operations, which are allocated to assembly devices in real time by inter-holon negotiation. The performance of this task allocation mechanism is validated in simulation. Experimental results on a real assembly system are also presented.

The authors consider a stable slow-speed drive and high-precision positioning mechanism, applying a linear voltage drive to the brushless DC motor. To compare the performance of the linear voltage drive and switch drive, the brushless DC motor and two driving circuits have been modeled, and computer simulations have been performed by using MATRIXx. The validity of the slow-speed brushless DC motor control system was also proved by experiment. According to the experimental results, a rotational speed fluctuation can be obtained within 1%, where the rotational speed is 2 rad/sec and the given load is 0.3 N-m. It was verified that the performance of the positioning mechanism is sufficiently precise by testing the response to a step load. >

We have developed two types of micro mobile machine with wheels made up of two electromagnetic ultrasmall motors and four wheels. One is a wired type of size of less than 10 mm/spl times/10 mm/spl times/10 mm. The energy and control signal is supplied through the electric wire. It can be controlled by changing the frequency supplied to the two motors respectively in four directions: forward, backward, left-turn, and right-turn. It has achieved a speed of 300 mm/sec and a gradability of 15 degrees. The other is a wireless type of size of less than 1 in/spl times/1 in/spl times/1 in. It is installed with the sensor, the driving control circuit and battery. Wireless operation is made possible by optically transmitting infrared ray signals. It can drive in four directions on a horizontal plane. This paper describes these structures and how to control these machines.

We analyzed PyNPase expression discriminating between cancer and tumor stroma of the colorectum by Western blotting using a newly developed extraction method from microdissected tissue sections fixed with buffered formalin. Analysis of 98 colorectal cancers revealed that PyNPase was as high as 70.2 +/- 18.5 unit/mg protein in the stroma fraction (SF), whereas it was 45.1 +/- 10.5 in the cancer fraction (CF) (p0.0001). Vessel density was correlated with PyNPase in the SF but not in the CF. In stage IIIb, 11 cases expressing a high level of PyNPase in the CF showed poorer prognosis than 10 cases with low-level PyNPase expression (p0.05), although the level of PyNPase expression in the SF did not affected the patients prognosis. Immunohistochemical examination indicated that PyNPase in the SF was mainly produced by macrophages (M phi), and therefore we investigated the profile of PyNPase production by M phi. In in vitro experiments PyNPase production by M phi was greatly enhanced by stimulation with OK-432, and the culture supernatant had the ability to convert 5'DFUR to 5-FU.

To evaluate residual effects of inhalational anesthetics after reversal of neuromuscular blocking agent, neuromuscular function was monitored after halothane or sevoflurane anesthesia in thirty-seven patients (ASA physical status I or II) for elective surgery after obtaining informed consent. Electromyograph of the adductor pollicis muscle in response to train of four (TOF) stimulation was monitored throughout the study. The first twitch of TOF (T1; % of its control) and the ratio of the fourth twitch to the first twitch of TOF (T4/T1; TR) were recorded at 0, 2, 5, 10, and 15 min after reversal. The patients were divided into five groups; 1) the fentanyl group (n = 7) received fentanyl/N2O; 2) in the halothane stop group (n = 6), halothane was discontinued at least fifteen minutes before neostigmine administration; 3) in the halothane stable group (n = 7), 0.7% halothane was maintained until fifteen minutes after neostigmine; 4) in the sevoflurane stop group (n = 12), sevoflurane was discontinued fifteen minutes before the reversal; 5) in the sevoflurane stable group (n = 5), 3% sevoflurane was maintained until fifteen minutes after the reversal. Anesthesia was induced by thiopental 4 mg.kg-1 and suxamethonium 1 mg.kg-1 and the patients were intubated. After initial dose of vecuronium 0.1 mg.kg-1, the additional dose of 0.02 mg.kg-1 was administered to maintain T1 under 10% of the control value. At the end of the surgery atropine 0.015 mg.kg-1 and neostigmine 0.04 mg.kg-1 were administered to reverse vecuronium when T1 had recovered to 25% of its control. Halothane groups did not differ from fentanyl group. Recovery of T1 at 15 min was suppressed after discontinuation of sevoflurane (86.0 +/- 8.2%) in comparison with fentanyl (97.0 +/- 8.3%). Both T1 (75.4 +/- 12.2%) and TR (68.0 +/- 12.6%) at 15 min after the reversal during 3% sevoflurane inhalation were below those of the stable group. We conclude that the residual sevofulrane after discontinuation of inhalation may impair the neuromuscular transmission after the reversal of neuromuscular blockade. Neuromuscular function should be monitored after the end of anesthesia even though the patient is fully awake.

We investigated the effect of mild exercise on global and regional diastolic function in patients with hypertensive left ventricular hypertrophy (LVH), using simultaneous measurement of micromanometric left ventricular pressure and echocardiographic indexes. Although left ventricular end-diastolic pressure (LVEDP) at rest in the hypertensive LVH group was similar to that in the control normotensive group, who had no abnormality on echocardiography or cardiac catheterization, LVEDP showed a marked increase during exercise only in the hypertensive LVH group. The pressure-dimension relation was shifted upward during exercise in all patients with hypertensive LVH. We utilized a new index of regional diastolic myocardial stiffness, KDM, which is derived from the diastolic relation between the mean wall stress and the natural logarithm of the reciprocal of wall thickness. Although KDM at rest was similar in the two groups, KDM increased significantly during exercise, both in the septum and posterior wall in patients with hypertensive LVH. The present results imply that even low-grade exercise may have deleterious effects on regional myocardial performance in patients with hypertensive LVH.

The aim of this study was to clarify that the depressed mechanoenergetics in patients with idiopathic dilated cardiomyopathy (DCM) resulted from compensation for the decreased contractility. The study population consisted of eight control subjects, with normal left ventricular size and ejection fraction and 31 patients with DCM. Left ventricular end-systolic elastance (Ees), effective arterial elastance (Ea), external work (EW), and the pressure-volume area (PVA) were measured, using a dual-field volume conductance catheter equipped with a micromanometer-tipped catheter. Ea/Ees was evaluated as ventriculoarterial coupling. Normal hearts worked at almost optimal condition (defined as Ea/Ees = 1/2), while ventriculoarterial coupling was far from the optimum (EaEes) in patients with DCM. Ees in patients with DCM was less than that in control subjects; however, Ea was similar in the two groups. The mismatch of Ea/Ees observed in DCM leads to depressed mechanoenergetics as a result of the compensatory response to maintain adequate blood pressure. Volume enlargement plays an important role in maintaining adequate blood pressure and cardiac output in the course of chronic deterioration of contractility.

Forty-three sporadic gastric cancers were analyzed with regard to whether mutations of simple repeated sequences in the transforming growth factor beta type II receptor (TbetaR-II) gene are associated with microsatellite instability (MSI) and gastric carcinogenesis. In 12 of the 43 cancers (28%), MSI was observed at least at 1 of the 2 microsatellite loci. Frameshift mutations of the TbetaR-II gene, all of which were 1 base deletion of 10 adenine repeats, were detected in 3 of 6 cancers, with MSI at 2 loci. However, mutations were not detected in 6 cancers, with MSI only at 1 locus and 31 cancers without MSI. Moreover, microanalysis in these cases revealed that the mutant-type alleles of TbetaR-II were invariably common in different areas within the tumor, in contrast to the markedly variable alleles of microsatellite loci. Our results suggest that frameshift mutation of the TbetaR-II gene may be a critical event associated with MSI and may contribute to carcinogenesis of the stomach. One of the possible mechanisms of escape from growth control by TGFbeta during gastric carcinogenesis could involve frameshift mutations of the TbetaR-II gene caused by DNA replication errors.

pRB, the retinoblastoma tumor suppressor gene product, regulates the cell cycle at G1/S transition negatively. Many cell cycle regulators modulate pRB function through its phosphorylation status. G1 cyclins (cyclin D, E)/cyclin-dependent kinases (cdk2, 4) inactivates pRB through its phosphorylation, while p21 (WAF1) and p16 inhibit cdks. In several kinds of cancer, Rb gene alteration or functional inactivation of pRB has been reported. In esophageal cancer, loss of heterozygosity of Rb gene and cyclin D gene amplification were frequently detected. But in gastric and colorectal cancer, Rb gene loss or deletion has been shown to be rare. In this study we investigated the expression of pRB, G1 cyclins, cdks and cdk-inhibitors in adenoma-carcinoma sequence of colorectum. And we compared the phosphorylation status of pRB in colorectal normal mucosa and cancer tissue. In adenoma only cyclin D and E were overexpressed but not cdks. In cancer in adenoma pRB and cdk2 were overexpressed with high frequency, and cdk4 overexpression was detected in advanced cancer. p16 overexpression was detected in almost all cancers, but in contrast p21 overexpression was rare event. Comparative study showed that pRB-positive cancer cells also expressed both cdc2/cdk2 and cyclin E. Densitometric analysis revealed that in advanced cancer pRB was hyperphosphorylated compared with normal mucosa. These results indicate that overexpression of cyclin D/cdk4 and cyclin E/cdk2 would phosphorylate pRB, and insufficient expression of p21 may accelerate pRB inactivation.

G1/S transition of cell cycle is regulated by G1 cyclins/cyclin-dependent kinases (CDKs) and their inhibitors. Cyclin D/CDK4 complex phosphorylates retinoblastoma gene product and p16 inhibits CDK4 in competition with cyclin D. Aberrant expression of these proteins may deregulate cell proliferation and moreover may lead to tumor formation and progression. In this study we examined the expression of cyclin D, CDK4 and p16 in adenoma-carcinoma sequence of colorectum. Immunohistochemical staining revealed that cyclin D was overexpressed in 55 of 66 (83%) adenomas, 8 of 8 (100%) carcinomas in adenoma (CIAs), 26 of 45 (58%) advanced carcinomas and even in hyperplastic mucosa. CDK4 was overexpressed in 0 of 52 (0%) adenomas, 0 of 14 (0%) CIAs and 39 of 45 (87%) advanced carcinomas, but not in hyperplastic mucosa. p16 was overexpressed in only 5 of 52 (10%) adenomas, 10 of 14 (71%) CIA and 59 of 60 (98%) advanced carcinomas, but not in hyperplastic mucosa. And comparative study of CDK4 and p16 showed that CDK4-positive cancer cells also expressed p16. Western blot analysis also revealed that primary regions of advanced colorectal cancer tissues showed 4.61, 3.30, 1.65 and 8.03-fold overexpression on the average in cyclin D1, cyclin D2, CDK4 and p16 respectively. These results indicate that both cyclin D and CDK4 may contribute to phosphorylation of pRB. p16 overexpression would be induced as a brake at G1/S transition through pRB phosphorylation by CDK4 overexpression in advanced colorectal carcinomas.

The nucleotide sequence of the citC coding for the citrate carrier in several Salmonella serovars has been determined, and the amino acid sequence of the carrier protein was deduced. The predicted citrate carrier from Salmonella pullorum and Salmonella enteritidis consists of 446 amino acids with a molecular weight of 47,621, whereas that from Salmonella dublin is the same 446 amino acids with the slightly different molecular weight of 47,591, because 1 amino acid residue was substituted. The predicted proteins are highly hydrophobic (69% nonpolar amino acids). The hydropathy profile suggests that the proteins are composed of 11-12 hydrophobic membrane-spanning segments with two hydrophilic cores in the middle of the protein sequence. No homology in the nucleotide and amino acid sequences was found in the molecular structures of citA, citP, and tctI genes. The citC-coding citrate transport activity is Na(+)-dependent and specific for citrate only. The transcript from the citC gene was not detected in the total RNA from several Salmonella serovars except S. dublin in Northern blot analysis, although the promoter of the citC genes appeared to be functional in Escherichia coli and Salmonella typhimurium strains using the lacZ fusion assay. These results suggested that the citC gene-coding citrate carrier is probably a TctIII system such as that identified previously in S. typhimurium.

A cryptic citrate transport gene (citA) from Salmonella typhimurium chromosome was cloned and its nucleotide sequence was determined. The cloned plasmid conferred citrate-utilizing ability on wild-type Escherichia coli, which cannot grow on citrate as the sole source of carbon. The resultant E. coli transformant was able to transport citrate. A 1,302-base-pair open reading frame with a preceding ribosomal binding site was found in the cloned DNA fragment. The 434-amino-acid protein that could be translated from this open reading frame is highly hydrophobic (69% nonpolar amino acid residues), consistent with the fact that the transport protein is an intrinsic membrane protein. The molecular weight of this protein was calculated to be 47,188. The gene sequence determined is highly homologous to those of Cit+ plasmid-mediated citrate transport gene, citA, from E. coli, the chromosomal citA gene from Citrobacter amalonaticus and the chromosomal cit+ gene from Klebsiella pneumoniae. The hydropathy profile of the deduced amino acid sequence suggests that this carrier has 12 hydrophobic segments, which may span the membrane lipid bilayer.

The activation of the complement system of rainbow trout by trout C-reactive protein (CRP) was investigated. Complement fixation tests were performed by using rabbit hemolysin-sensitized sheep erythrocytes and rainbow trout complement. Purified CRP increased the consumption of complement in the presence of Streptococcus pneumoniae C-polysaccharide (CPS), indicating the activation of the complement system. In contrast to this, acute phase serum activated the complement in the absence of CPS. Consumption of the complement by acute-phase serum was depressed when CRP was removed from acute-phase serum by CPS-sepharose 4B affinity chromatography. The acute-phase serum, as well as CRP plus CPS, suppressed in vitro growth of Vibrio anguillarum in the presence of complement, and enhanced the phagocytosis of the bacteria by glass-adherent peritoneal exudate cells. These results indicated that CRP has a role in host defense during acute-phase response through the activation of the complement system, enhancement of phagocytosis, and suppression of bacterial growth.