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Sclerosing angiomatoid nodular transformation (SANT) is a rare benign vascular disease and mostly discovered incidentally in asymptomatic patients. Since SANT grows over time and it is often difficult to rule out malignancy on imaging, splenectomy is frequently the treatment of choice. Image findings of SANT have been reported as low signal intensity on T2-weighted magnetic resonance (MR) images and a characteristic enhancement pattern on dynamic contrast-enhanced computed tomography (CT) and MR images. This report describes a case of a 43-year-old woman with SANT presenting multiple calcifications on CT images, and discusses the characteristics and mechanism of the calcification, while reviewing previously reported cases of SANT., (© 2024 The Authors. Published by Elsevier Inc. on behalf of University of Washington.)

Antimicrobial-resistant bacteria, selected by antimicrobial agent use in livestock, are emerging and their spread to crops from feces via composting represents a public health concern as they are ultimately transmitted to humans. In this study, we investigated Escherichia coli and other ampicillin (AMP)-resistant coliform spread conditions in field soil and dent corn, an agricultural crop, on a livestock-derived compost-applying farm. No AMP-resistant E. coli was detected in any samples of field soil and dent corn. In contrast, AMP-resistant and extended-spectrum β-lactam (ESBL) producing coliforms were consistently present in field soil and dent corn during the entire study period. In particular, extremely high AMP-resistant coliform levels were detected in dent corn stems and roots. AMP-resistant coliform detection in crops is pivotal and raises significant concerns regarding antimicrobial-resistant bacterial spread. Furthermore, AMP-resistant coliform isolate identification defined Enterobacter bugandensis and Enterobacter asburiae as the dominant species among AMP-resistant coliforms, both tested positive for ESBL production. This means that high concentrations of AMP-resistant coliforms are to be present on farms where crops are grown. However, we identified no common species among the AMP-resistant coliforms in the compost, field soil, and dent corn samples. Therefore, the initial hypothesis of the compost being the source of antimicrobial-resistant bacteria was not confirmed. Although their source remains unknown, a certain antimicrobial-resistant bacterial concentration could nonetheless be detected in the field., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Understanding the current situation and risk of environmental contamination by anti-influenza drugs in aquatic environments is key to prevent the unexpected emergence and spread of drug-resistant viruses. However, few reports have been focused on newer drugs that have recently been introduced in clinical settings. In this study, the behaviour of the prodrug baloxavir marboxil (BALM)-the active ingredient of Xofluza, an increasingly popular anti-influenza drug-and its pharmacologically active metabolite baloxavir (BAL) in the aquatic environment was evaluated. Additionally, their presence in urban rivers and a wastewater treatment plant (WWTP) in the Yodo River basin was investigated and compared with those of the major anti-influenza drugs used to date (favipiravir (FAV), peramivir (PER), laninamivir (LAN), and its active metabolite, laninamivir octanoate (LANO), oseltamivir (OSE), and its active metabolite, oseltamivir carboxylate (OSEC), and zanamivir (ZAN)) to comprehensively assess their environmental fate in the aquatic environment. The results clearly showed that BALM, FAV, and BAL were rapidly degraded through photolysis (2-h, 0.6-h, and 0.4-h half-lives, respectively), followed by LAN, which was gradually biodegraded (7-h half-life). In addition, BALM and BAL decreased by up to 47 % after 4 days and 34 % after 2 days of biodegradation in river water. However, the remaining conventional drugs, except for LANO (<1 % after 10 days), were persistent, being transported from the upstream to downstream sites. The LogK d values for the rates of sorption of BALM (0.5-1.6) and BAL (1.8-3.1) on river sediment were higher than those of conventional drugs (-0.5 to 1.7). Notably, all anti-influenza drugs were effectively removed by ozonation (>90-99.9 % removal) after biological treatment at a WWTP. Thus, these findings suggest the importance of introducing ozonation to reduce pollution loads in rivers and the environmental risks associated with drug-resistant viruses in aquatic environments, thereby promoting safe river environments., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests or personal relationships that could have influenced the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a great public health problem and is associated with many disease outbreaks and high mortality rates. Alarmingly, K. pneumoniae has been isolated from food in several recent studies. This study aimed to investigate the prevalence and characteristics of CRKP in food samples from Egypt., Methods: A total of 311 food samples (including 116 minced meat, 92 chicken meat, 75 diced meat, and 28 mutton) were collected from local markets in Egypt and were screened for CRKP with the determination of their antimicrobial resistance profiles. The whole genome sequence was done for 23 CRKP isolates to clarify the relationship between CRKP from food and human cases in Egypt using the SNP core genome. The conjugation probability of the bla NDM-5 harboring plasmid was identified using oriTfinder RESULTS: CRKP was isolated from 11% (35/311) of the samples, with 45.71% (16/35) of them showing resistance to colistin, one of the last-resort options for treating CRKP-mediated infections. In addition to the carbapenem and colistin resistance, the CRKP isolates frequently exhibited resistance to multiple antimicrobials including β-lactams, fluoroquinolones, aminoglycosides, tetracyclines, and chloramphenicol. In addition, most of the CRKP were potentially hypervirulent K. pneumoniae (HvKP) identified as phylogroup Kp1 and of high-risk groups as detected in STs reported in many human outbreaks globally, such as ST383 and ST147. The core-genome phylogeny showed similarities between the isolates from this study and those previously isolated from clinical human samples in Egypt. In addition, analysis of the plasmid on which bla NDM is encoded revealed that several antimicrobial resistance genes such as bla OXA-9 , bla CTX-M-15 , aac(6')-Ib, qnrS1, and several virulence genes are encoded on the same plasmid., Conclusions: This study is significant for food safety and public health and is important to further identify the change in the epidemiology of CRKP infections, especially the consumption of contaminated food products., Competing Interests: Declaration of Competing Interest There are no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The emergence and spread of antimicrobial resistance (AMR) continue on a global scale. The impacts of wastewater on the environment and human health have been identified, and understanding the environmental impacts of hospital wastewater and exploring appropriate forms of treatment are major societal challenges. In the present research, we evaluated the efficacy of ozone (O 3 )-based advanced wastewater treatment systems (O 3 , O 3 /H 2 O 2 , O 3 /UV, and O 3 /UV/H 2 O 2 ) for the treatment of antimicrobials, antimicrobial-resistant bacteria (AMRB), and antimicrobial resistance genes (AMRGs) in wastewater from medical facilities. Our results indicated that the O 3 -based advanced wastewater treatment inactivated multiple antimicrobials (>99.9%) and AMRB after 10-30 min of treatment. Additionally, AMRGs were effectively removed (1.4-6.6 log 10 ) during hospital wastewater treatment. The inactivation and/or removal performances of these pollutants through the O 3 /UV and O 3 /UV/H 2 O 2 treatments were significantly (P < 0.05) better than those in the O 3 and O 3 /H 2 O 2 treatments. Altered taxonomic diversity of microorganisms based on 16S rRNA gene sequencing following the O 3 -based treatment showed that advanced wastewater treatments not only removed viable bacteria but also removed genes constituting microorganisms in the wastewater. Consequently, the objective of this study was to apply advanced wastewater treatments to treat wastewater, mitigate environmental pollution, and alleviate potential threats to environmental and human health associated with AMR. Our findings will contribute to enhancing the effectiveness of advanced wastewater treatment systems through on-site application, not only in wastewater treatment plants (WWTPs) but also in medical facilities. Moreover, our results will help reduce the discharge of AMRB and AMRGs into rivers and maintain the safety of aquatic environments., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests or personal relationships that may have influenced the work reported in this study., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Balloon-Assisted Ultrasound-Guided Percutaneous Thrombin Injection (BATI) is useful for iatrogenic pseudoaneurysm. In previous reports, BATI for pseudoaneurysm of the femoral artery was performed by contralateral transfemoral approach. It has been reported that patients with obesity, hypertension, and diabetes have more hemorrhagic complications at the puncture site of the femoral artery. We report a case in which BATI by transradial artery approach was useful for patients with obesity, hypertension, and diabetes who were expected to be at high risk of hemorrhagic complications of transfemoral artery approach., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Occurrence of profiles of the carbapenem-resistant Escherichia coli (CRE-E) and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli (ESBL-E) in an urban river in a sub-catchment of the Yodo River Basin, one of the representative water systems of Japan was investigated. We conducted seasonal and year-round surveys for the antimicrobial-resistant bacteria (AMRB) and antimicrobial-resistance genes (AMRGs) in hospital effluents, sewage treatment plant (STP) wastewater, and river water; subsequently, contributions to wastewater discharge into the rivers were estimated by analyses based on the mass flux. Furthermore, the characteristics of AMRB in the water samples were evaluated on the basis of antimicrobial susceptibility tests. CRE-E and ESBL-E were detected in all water samples with mean values 11 and 1900 CFU/mL in the hospital effluent, 58 and 4550 CFU/mL in the STP influent, not detected to 1 CFU/mL in the STP effluent, and 1 and 1 CFU/mL in the STP discharge into the river, respectively. Contributions of the pollution load derived from the STP effluent discharged into the river water were 1 to 21%. The resistome profiles for bla IMP , bla TEM , and bla CTX-M genes in each water sample showed that AMRGs were not completely removed in the wastewater treatment process in the STP, and the relative abundances of bla IMP , bla TEM , and bla CTX-M genes were almost similar (P<0.05). Susceptibility testing of antimicrobial-resistant E. coli isolates showed that CRE-E and ESBL-E detected in wastewaters and river water were linked to the prevalence of AMRB in clinical settings. These results suggest the importance of conducting environmental risk management of AMRB and AMRGs in the river environment. To our knowledge, this is the first detailed study that links the medical environment to CRE-E and ESBL-E for evaluating the AMRB and AMRGs in hospital effluents, STP wastewater, and river water at the basin scale on the basis of mass flux as well as the contributions of CRE-E and ESBL-E to wastewater discharge into the river., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In situ continuous glucose monitoring under physiological culture conditions is imperative in understanding the dynamics of cell and tissue behaviors and their physiological responses since glucose plays an important role in principal source of biological energy. We therefore examined physiologically relevant dynamic changes in glucose levels based on glucose metabolism and production during aerobic culture (10% O 2 ) of rat primary hepatocytes stimulated with insulin or glucagon on a highly O 2 permeable plate, which can maintain the oxygen concentration close to the periportal zone of the liver. As glucose monitoring devices, we used oxygen-independent glucose dehydrogenase-modified single-walled carbon nanotube electrodes placed close to the surface of the hepatocytes. The current response of glucose oxidation slightly decreased after the addition of insulin in the presence of glucose due to the acceleration of glucose uptake by the hepatocytes, whereas that significantly increased after the addition of glucagon and fructose even in the absence of glucose due to the conversion of fructose to glucose based on gluconeogenesis. These phenomena might be consistent relatively with the physiological behaviors of hepatocytes in the periportal region. The present monitoring system would be useful for the studies of glucose homeostasis and diabetes in vitro., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Periodontal disease is an inflammatory disease caused by periodontopathogenic bacteria, which eventually leads to bone tissue (alveolar bone) destruction as inflammation persists. Periodontal tissues have an immune system against the invasion of these bacteria, however, due to the persistent infection by periodontopathogenic bacteria, the host innate and acquired immunity is impaired, and tissue destruction, including bone tissue destruction, occurs. Osteoclasts are essential for bone destruction. Osteoclast progenitor cells derived from hematopoietic stem cells differentiate into osteoclasts. In addition, bone loss occurs when bone resorption by osteoclasts exceeds bone formation by osteoblasts. In inflammatory bone disease, inflammatory cytokines act on osteoblasts and receptor activator of nuclear factor-κB ligand (RANKL)-producing cells, resulting in osteoclast differentiation and activation. In addition to this mechanism, pathogenic factors of periodontal bacteria and mechanical stress activate osteoclasts and destruct alveolar bone in periodontitis. In this review, we focused on the mechanism of osteoclast activation in periodontitis and provide an overview based on the latest findings., (© 2021 The Authors.)

Background: Distal embolism is a frequent complication in percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI) due to thrombotic lesions. Distal embolism causes no reflow phenomenon, which leads to worse patient prognosis after AMI. There is no established treatment to prevent distal embolism in PCI for thrombotic lesions. The aim of this study is to investigate the usefulness of long inflation balloon angioplasty (LIBA) with perfusion balloon in PCI for AMI due to thrombotic lesions., Methods and Results: This is a case series study. We investigated 10 cases treated with LIBA for cases with massive thrombus remaining after thrombus aspiration therapy in primary PCI for acute myocardial infarction. We investigated the success rate of the procedure, residual stenosis rate, TIMI flow grade, TIMI frame count, and myocardial blush score in 10 cases of primary PCI with LIBA at our hospital. In all 10 cases, distal embolism was not observed by angiogram after LIBA. In 9 cases, residual stenosis was improved to less than 25% and the procedure was completed without a stent. Before PCI, all cases had TIMI flow grade 0, but in all 10 cases, TIMI flow grade 3 was obtained after PCI. The mean TIMI frame count was 19.6 ± 2.50 for RCA lesions and 27.5 ± 1.5 for LAD lesions. Regarding myocardial blush score, grade 3 was obtained in 8 cases and grade 2 was obtained in 2 cases., Conclusion: LIBA using a perfusion balloon is a useful technique in thrombus lesion to prevent distal embolism., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest. The corresponding author and co-author have not received any funding or device assistance for this study., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Introduction: In contrast to the study in other part of the world, information about characteristics of plasmids carrying antimicrobial resistance genes (ARGs) in Enterobacteriaceae derived from environmental water in tropical Asian countries including Thailand is limited. This study, therefore, aimed to gain insight into genetic information of antimicrobial resistance in environmental water in Thailand., Methods: Coliform bacteria were isolated from environmental water collected at 20 locations in Thailand and identified. Then, susceptibility profiles to ampicillin, cefazoline, cefotaxime, kanamycin, ciprofloxacin, sulfamethoxazole, tetracycline, and nalidixic acid were assessed. In addition, antimicrobial resistant genes integrons, and replicon types were analyzed. And furthermore, plasmids carrying bla TEM and tetM were identified by S1-PFGE analysis and confirmed transmissibility by transconjugation experiments., Results: In 130 coliform bacteria isolated, 89 were resistant to cefazoline while 41 isolates were susceptible. Cefazoline-resistant coliform bacteria were found to be significantly resistant to cefotaxime and tetracycline as compared to susceptible isolates. Hence, bla TEM and tetM correlating with β-lactam antibiotics and tetracycline, respectively, were analyzed found to co-localize on the IncFrepB plasmids in isolates from pig farms' wastewater by S1-PFGE analysis. And furthermore, transmissibility of the plasmids was confirmed., Conclusions: Results obtained in this study suggested that ARGs in coliform bacteria may have been spreading on the farm via IncFrepB plasmids. Hence, appropriate use of antimicrobials and good hygiene management on the farm are required to prevent the emergence and spread of resistant bacteria., (Copyright © 2021 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Background: An increase in the rate of relative poverty and the number of welfare recipients is a serious social problem in Japan. A recent overseas survey demonstrated that lack of health insurance was associated with increased in-hospital mortality in patients with ST-segment elevation myocardial infarction (STEMI). The aim of this study is to investigate the clinical features of STEMI patients who receive welfare public assistance in Japan., Methods: We enrolled 525 STEMI patients who were hospitalized in our hospital from 2010 to 2019. We divided patients into groups of patients receiving welfare public assistance (WPA group, N = 67) and groups of patients not receiving welfare public assistance (non-WPA group, N = 458). Patient characteristics, clinical outcome, and cardiac function on transthoracic echocardiography were compared., Results: WPA group were younger than non-WPA group (61.2 ± 10.9 years VS 64.5 ± 13.3 years, p = 0.03). The prevalence of smoking was higher in WPA group compared to non-WPA group (91.0% VS 81.1% p = 0.04) and high-density lipoprotein cholesterol value of WPA group was lower than in non-WPA group (43.2 ± 9.9 mg/dl vs 47.1 ± 12.8 mg/dl, p = 0.005). Ventricular arrhythmia on admission was significantly more frequent in WPA group (11.9% VS 4.8%, p = 0.02). In acute and chronic phase, left ventricular ejection fraction in WPA group was lower than non-WPA group (in acute phase 46.6 ± 10.7% vs 53.3 ± 8.6% p = 0.001, in chronic phase 48.7 ± 10.1% vs 55.3 ± 9.4%, p = 0.008)., Conclusion: STEMI patients receiving welfare public assistance had poorer control of coronary risk, increased risk of fatal arrhythmia, and reduced systolic function than those not receiving welfare public assistance. It is necessary to have a system that can strengthen lifestyle management, such as diet and smoking cessation for the purpose of improving the prognosis of welfare recipients after AMI., (Copyright © 2020 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.)

Clostridioides difficile infections (CDIs) are predominantly a healthcare-associated illness in developed countries, with the majority of cases being elderly and hospitalize patients who used antibiotic therapy. Recently, the incidence of community-associated CDIs (CA-CDIs) in younger patients without a previous history of hospitalization or antibiotic treatment has been increasing globally. C. difficile is sometimes found in the intestine of many animals, such as pigs, calves, and dogs. Food products such as retail meat products and vegetables sometimes contain C. difficile. C. difficile has also been isolated from several environments such as compost manure, rivers, and soils. Yet, direct transmission of C. difficile from animals, food products, and environments to humans has not been proven, although these strains have similar molecular characteristics. Therefore, it has been suggested that there is a relationship between CA-CDIs and C. difficile from animals, food products, and the environment. To clarify the importance of the presence of C. difficile in several sources, characterization of C. difficile in these sources is required. However, the epidemiology of C. difficile in animals, food products, and the environment is not well studied in Japan. This review summarizes recent trends of CDIs and compares the molecular characteristics of C. difficile in Japanese animals, food products, and the environment. The prevalence trends of C. difficile in Japan are similar to those in the rest of the world. Therefore, I recommend using a One Health approach to CDI surveillance, monitoring, and control., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Extended-spectrum β-lactamase (ESBL)-producing bacteria are spreading rapidly, posing a threat to human and animal health. Contamination of vegetables with antimicrobial-resistant bacteria or those harboring antimicrobial resistance genes or a combination of both presents a potential route of transmission to humans. Therefore, the aim of this study was to determine the prevalence of these bacteria in fresh vegetables in Japan. A total of 130 samples of fresh vegetables were collected from seven supermarkets in Japan. The predominant genus detected was Pseudomonas spp., including 10 ESBL-producing strains, isolated from 10 (7.7%) of the vegetable samples. Two ESBL genes were detected, bla TEM-116 ( n = 7) and bla SHV-12 ( n = 3), and some of these strains were resistant to multiple antibiotics. Because vegetables are often consumed raw, those contaminated with ESBL producers could represent an important route of transmission to humans in Japan. Thus, more stringent hygiene measures and monitoring are required to prevent transmission via this source.

The aim of the present study was to determine and compare the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and their molecular characteristics among veterinary staff in Sapporo in 2008 and 2016. We isolated MRSA from veterinarians (Vet; n = 91), veterinary technicians (VT; n = 113), and other staff members (n = 24) from 45 small animal hospitals (animal hospitals), as well as from surface swabs (n = 123) obtained from 37 animal hospitals, in 2016. MRSA was observed in 14 Vets (15%), 7 VTs (6%), 2 other staff members (8%), and 6 environmental samples (5%). The prevalence of MRSA among veterinary staff tended to decrease, in comparison to 2008. All the MRSA isolates were classified as CC5/SCCmecII, which is commonly observed in medical settings in Japan. Upon performing pulse-field gel electrophoresis, with SmaI and EagI, and clfB sequence typing, it was observed that 16 of the MRSA isolates from 2016 were highly similar to those obtained in 2008. This suggests that some MRSA isolates persisted throughout 8 years, although their origins remain unclear. The continuation of education and monitoring of MRSA is necessary for the prevention and control of infection in these settings., (Copyright © 2018 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Niemann-Pick disease type C (NPC) is a rare, progressive autosomal recessive disease. It is caused by mutations in either the NPC1 or NPC2 genes, resulting in defective regulation of intracellular lipid trafficking. Miglustat, which reversibly inhibits glucosylceramide synthase, reportedly has beneficial effects on the progressive neurological symptoms of NPC and was approved in Japan in 2012. Some reports suggested that miglustat therapy delayed the onset or progression of NPC when treatment was initiated before the onset of neurological manifestation or at an early stage. We report here a patient with the early-infantile form of NPC who started on miglustat at 4months of ages. To our knowledge, this patient is the youngest reported patient with NPC in which miglustat therapy was initiated. Our patient, who had hypotonia and developmental delay before treatment, remained stable and showed no new neurological symptoms. In addition, pulmonary involvement was improved during miglustat therapy. Our case and previous reports underscore the importance of early initiation of miglustat therapy for NPC., (Copyright © 2017 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

In this research, we analyzed the main sequence types (ST) and ST complexes of human- and chicken-derived isolates of Campylobacter jejuni in Japan by using multilocus sequence typing (MLST). We also analyzed lipooligosaccharide biosynthesis locus classes (LOS locus classes) and the numbers of isolates carrying genes coding resistance factors against various antibiotics, and observed their relationships. ST-21 complex was the main ST complex in isolates from humans (n = 38) and chickens (n = 25). None of the isolates showed resistance to imipenem, chloramphenicol, or erythromycin. Few isolates were resistant to ampicillin and streptomycin (1.3%-15%), whereas many showed resistance to tetracycline, ciprofloxacin, and nalidixic acid (38%-48%). Among the ST-21 complex isolates, ST4526 was detected at a very high rate. Those isolates showed resistance to tetracycline and ciprofloxacin, and were susceptible to ampicillin. Among the chicken-derived isolates, 37 of the 38 isolates that showed resistance to ciprofloxacin and nalidixic acid had threonine to isoleucine amino acid substitution in GyrA at codon 86 (T86I). Among the human-derived isolates, 17 of the 47 isolates that showed resistance to ciprofloxacin and 16 of the 48 isolates that showed resistance to nalidixic acid did not have T86I amino acid mutations in GyrA. The human-derived ST-21 complex isolates were classified into LOS locus classes A, B, C, D, and E. The chicken-derived ST-21 complex isolates, with the exception of one isolate, were all classified into LOS locus classes C and D. Among chicken-derived isolates, the most prevalent was ST51 (ST-443 complex) (10 isolates) and all of those were LOS locus class E., (Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

We describe a novel technology for detecting nucleic acids: Probe Alteration Link Self-Assembly Reactions (PALSAR). PALSAR comprises DNA self-assembly of pairs of short DNA probes formed by alternate hybridization of three complementary regions in a pair of honeycomb probes (HCPs). Self-assembly occurs at designated salt concentrations and reaction temperatures and requires no enzymes. We prepared pairs of HCPs to detect mRNAs encoded by the GAPDH gene β-actin (BA) gene, CD3D gene, CD4 gene, major vault protein (MV) gene and the signalling lymphocytic activation molecule-associated protein (SAP) gene, and succeeded in quantitatively detecting these mRNAs. PALSAR could detect mRNA directly without synthesizing cDNA. Moreover, multiple mRNAs could be detected simultaneously in a single reaction tube and there was a good correlation between the results obtained PALSAR and those by real-time PCR., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)

Quinolones have long been used as the first-line treatment for Campylobacter infections. However, an increased resistance to quinolones has raised public health concerns. The development of new quinolone-based antibiotics with high activity is critical for effective, as DNA gyrase, the target of quinolones, is an essential enzyme for bacterial growth in several mechanisms. The evaluation of antibiotic activity against Campylobacter jejuni largely relies on drug susceptibility tests, which require at least 2 days to produce results. Thus, an in vitro method for studying the activity of quinolones against the C. jejuni DNA gyrase is preferred. To identify potent quinolones, we investigated the interaction of C. jejuni DNA gyrase with a number of quinolones using recombinant subunits. The combination of purified subunits exhibited DNA supercoiling activity in an ATP dependent manner. Drug concentrations that inhibit DNA supercoiling by 50% (IC50s) of 10 different quinolones were estimated to range from 0.4 (sitafloxacin) to >100 μg/mL (nalidixic acid). Sitafloxacin showed the highest inhibitory activity, and the analysis of the quinolone structure-activity relationship demonstrated that a fluorine atom at R-6 might play the important role in the inhibitory activity against C. jejuni gyrase. Measured quinolone IC50s correlated well with minimum inhibitory concentrations (R = 0.9943). These suggest that the in vitro supercoiling inhibition assay on purified recombinant C. jejuni DNA gyrase is a useful and predictive technique to monitor the antibacterial potency of quinolones. And furthermore, these data suggested that sitafloxacin might be a good candidate for clinical trials on campylobacteriosis., (Copyright © 2015 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Phenotypic observation of thioredoxin peroxidase-1 (TPx-1) gene-disrupted Plasmodium berghei (TPx-1 KO) in the liver-stage was performed with an in vitro infection system in order to investigate defective liver-stage development in a mouse infection model. Indirect immunofluorescence microscopy assay with anti-circumsporozoite protein antibody revealed that in the liver schizont stage, TPx-1 KO parasite cells were significantly smaller than cells of the wild-type parent strain (WT). Indirect immunofluorescence microscopy assay with anti-merozoite surface protein-1 antibody, which was used to evaluate late schizont-stage development, indicated that TPx-1 KO schizont development was similar to WT strain development towards the merozoite-forming stage (mature schizont). However, fewer merozoites were produced in the mature TPx-1 KO schizont than in the mature WT schizont. Taken together, the results suggest that TPx-1 may be involved in merozoite formation during liver schizont development., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Resistance to broad-spectrum cephalosporins (BSCs) in Enterobacteriaceae in companion animals has become a great concern for public health. To estimate the dissemination of BSC-resistant bacteria between dog and human, we examined the BSC-resistance determinants of and genetic similarities between 69 BSC-resistant Escherichia coli isolates derived from canine rectal swabs (n = 28) and human clinical samples (n = 41). Some E. coli isolates possessed blaTEM-1b (14 canine and 16 human isolates), blaCTx-M-2 (6 human isolates), blaCTx-M-14 (3 canine and 14 human isolates), blaCTx-M-27 (1 canine and 15 human isolates), and blaCMY-2 (11 canine and 3 human isolates). The possession of CTX-M-type β-lactamases was significantly more frequent in human isolates, whereas CMY-2 was more common in canine isolates. Bacterial typing methods (phylogenetic typing, O-antigen serotyping, and pulsed-field gel electrophoresis) showed little clonal relationship between canine isolates and human isolates. Plasmid analysis and Southern blotting indicated that the plasmids encoding CMY-2 were similar among canine and human isolates. Based on the differences in the major β-lactamase and the divergence of bacterial types between canine and human isolates, it seems that clonal dissemination of BSC-resistant E. coli between canines and humans is limited. The similarity of the CMY-2-encoding plasmid suggests that plasmid-mediated β-lactamase gene transmission plays a role in interspecies diffusion of BSC-resistant E. coli between dog and human., (Copyright © 2013 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

mRNA and protein expression profiles for three peroxiredoxins (TPx-1, TPx-2 and 1-Cys Prx) of liver stage Plasmodium berghei were examined through quantitative reverse transcription-PCR (RT-PCR) and indirect immunofluorescence microscopy assay (IFA). RT-PCR experiments revealed that mRNA expression for the TPx-1 was detected shortly after the sporozoite infection and kept expressed until the schizont stage. In contrast, the mRNA expression for 1-Cys Prx had begun increasing when the parasite developed into the schizont stage. Using the IFA, TPx-1 and 1-Cys Prx were detected in the cytosol. This finding suggested the developmental stage-specific expression of the cytosolic enzymes in the liver stage parasite. On the other hand, the mRNA expression for TPx-2 had begun increasing at the trophozoite stage and peaked at the schizont stage. In the IFA, TPx-2 was found localized in the mitochondria. The increase of TPx-2 might be explained by the exponential development of the parasite during the schizont stage requiring ATP production which may induce reactive oxygen species (ROS) in the mitochondria., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Wheat (Triticum aestivum) and rice (Oryza sativa) are two of the most agriculturally important cereal crop plants. Rice is known to produce numerous diterpenoid natural products that serve as phytoalexins and/or allelochemicals. Specifically, these are labdane-related diterpenoids, derived from a characteristic labdadienyl/copalyl diphosphate (CPP), whose biosynthetic relationship to gibberellin biosynthesis is evident from the relevant expanded and functionally diverse family of ent-kaurene synthase-like (KSL) genes found in rice the (OsKSLs). Herein reported is the biochemical characterization of a similarly expansive family of KSL from wheat (the TaKSLs). In particular, beyond ent-kaurene synthases (KS), wheat also contains several biochemically diversified KSLs. These react either with the ent-CPP intermediate common to gibberellin biosynthesis or with the normal stereoisomer of CPP that also is found in wheat (as demonstrated by the accompanying paper describing the wheat CPP synthases). Comparison with a barley (Hordeum vulgare) KS indicates conservation of monocot KS, with early and continued expansion and functional diversification of KSLs in at least the small grain cereals. In addition, some of the TaKSLs that utilize normal CPP also will react with syn-CPP, echoing previous findings with the OsKSL family, with such enzymatic promiscuity/elasticity providing insight into the continuing evolution of diterpenoid metabolism in the cereal crop plant family, as well as more generally, which is discussed here., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Two of the most agriculturally important cereal crop plants are wheat (Triticum aestivum) and rice (Oryza sativa). Rice has been shown to produce a number of diterpenoid natural products as phytoalexins and/or allelochemicals--specifically, labdane-related diterpenoids, whose biosynthesis proceeds via formation of an eponymous labdadienyl/copalyl diphosphate (CPP) intermediate (e.g., the ent-CPP of gibberellin phytohormone biosynthesis). Similar to rice, wheat encodes a number of CPP synthases (CPS), and the three CPS characterized to date (TaCPS1-3) all have been suggested to produce ent-CPP. However, several of the downstream diterpene synthases will only react with CPP intermediate of normal or syn, but not ent, stereochemistry, as described in the accompanying report. Investigation of additional CPS did not resolve this issue, as the only other functional synthase (TaCPS4) also produced ent-CPP. Chiral product characterization of all the TaCPS then established that TaCPS2 uniquely produces normal, rather than ent-, CPP, thus, providing a suitable substrate source for the downstream diterpene synthases. Notably, TaCPS2 is most homologous to the similarly stereochemically differentiated syn-CPP synthase from rice (OsCPS4), while the non-inducible TaCPS3 and TaCPS4 cluster with the rice OsCPS1 required for gibberellin phytohormone biosynthesis, as well as with a barley (Hordeum vulgare) CPS (HvCPS1) that also is characterized here as similarly producing ent-CPP. These results suggest that diversification of labdane-related diterpenoid metabolism beyond the ancestral gibberellins occurred early in cereal evolution, and included the type of stereochemical variation demonstrated here., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: Surgical treatment of few complex pituitary adenomas, mostly large/giant and multilobulated, still remains troublesome. We present our case series using a simultaneous combined supra-infrasellar approach for these tumors., Methods: We retrospectively reviewed our 29 consecutive patients who have undergone procedures using this approach since 2004. Preoperative characteristics and surgical outcomes were reviewed., Results: Total removal of the adenoma was achieved in 7 cases, subtotal removal in 21 cases, and partial removal in 1 case. Although surgical complications apart from pituitary insufficiency were noted in 12 patients, most were transient but permanent oculomotor nerve palsy and worsening of preoperative visual disturbance developed in 2 and 1 patient, respectively., Conclusions: Because these complex adenomas are not only large and invasive but also tend to lack a well-defined tumor capsule, the combined approach was effective for the accurate and safe removal by manipulating from both sides. The present single-stage approach is a viable option for these few selected patients., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Periodontitis is characterized by chronic gingival tissue inflammation, and inflammatory mediators such as IL-8 and prostaglandin E(2) (PGE(2)) are associated with disease progression. Previously we showed that oxidatively modified low-density lipoprotein (oxLDL) was present in gingival crevicular fluid. In this study, the role of oxLDL in the gingival epithelial cell inflammatory response was further investigated using Ca9-22 cells and primary human oral keratinocytes (HOK). Treatment of Ca9-22 cells and HOK with oxLDL induced an up-regulation of IL-8 and the PGE(2)-producing enzymes, cyclooxygenase-2 and microsomal PGE(2) synthase-1. These responses induced by oxLDL were significantly suppressed by a nuclear factor-kappa B (NF-κB) inhibitor. However, unlike the result in macrophages, oxLDL did not lead to an increase in CD36 expression in these two cells. These results suggest that oxLDL elicits gingival epithelial cell inflammatory responses through an activation of the NF-κB pathway. These data suggest a mechanistic link between periodontal disease and lipid metabolism-related disorders, including atherosclerosis., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Background: Osteosarcoma is the most frequent primary malignant bone tumor. In Europe and the United States, its prognosis has been greatly improved by the use of multimodal treatment, including preoperative and postoperative chemotherapy as well as surgery. In Japan, however, only a few clinical studies on osteosarcoma have been carried out., Methods: To evaluate the efficacy of neoadjuvant chemotherapy on nonmetastatic, operable osteosarcoma arising in the extremities, a prospective multi-institutional phase II trial, the Neoadjuvant Chemotherapy for Osteosarcoma (NECO) study, was conducted. Preoperative chemotherapy included high-dose methotrexate (HD-MTX), cisplatin (CDDP), and adriamycin (ADR). If the induction therapy was assessed as not effective, high-dose ifosfamide (IFO) was added to the chemotherapy regimen. A total of 124 patients were enrolled in this trial, and ultimately 113 patients were eligible., Results: The 5-year overall survival (OAS) and event-free survival (EFS) rates in the NECO study were 77.9% and 65.5%, respectively. A good histological response to the induction chemotherapy resulted in favorable OAS (78.7%). The patients assessed as poor histological responders with progressive disease after the induction chemotherapy exhibited comparable outcomes (OAS 89.5%, EFS 68.2%). There were no significant differences between the OAS and EFS rates of the patients in terms of response to preoperative chemotherapy., Conclusions: We analyzed the results of the intensive neoadjuvant chemotherapy and the effects of adding IFO on patients with osteosarcoma in Japan. The results suggest efficacy of the high-dose IFO addition to the standard three-drug chemotherapy regimen. However, a randomized clinical study is needed to establish the true impact of IFO on patients with osteosarcoma.

Purpose: Although rare, hand injury caused by puncture with the sea urchin spine can result in serious complications. To emphasize its clinical significance, this article describes a group of patients who sustained chronic granulomatous arthritis induced by puncture with sea urchin spine (designated sea urchin spine arthritis)., Methods: Five patients who developed sea urchin spine arthritis of the hand after puncture with sea urchin spine were treated at our hospitals. All lesions involved the proximal interphalangeal (PIP) joint (4 index fingers and 1 middle finger). Patients experienced pain, swelling, and discomfort around the site of puncture immediately after the injury. These initial symptoms subsided within a few days, and secondary symptoms including fusiform swelling, limited motion, and mild pain of the PIP joint appeared from 1 to 2 months later. Laboratory tests of inflammation and blood cell counts were negative. Plain radiographs showed soft tissue swelling and osteolysis but no visible spine. Thorough synovectomy of the PIP joint was performed, and the granulation tissue around the joint was also removed., Results: No microorganism was identified from tissue culture or polymerase chain reaction in any of the 5 patients. At a mean follow-up of 21 months, 2 patients exhibited essentially normal active motion of the affected PIP joint, whereas the remaining 3 patients had diminished range of motion., Conclusions: Diagnosis of sea urchin spine arthritis can be made by history of sea urchin spine injury, a symptom-free period before the development of synovitis, and the absence of laboratory test abnormalities. Neither antibiotics nor nonsteroidal anti-inflammatory agents are effective. Undertaken early enough, thorough synovectomy might avoid complications and obtain favorable results., Type of Study/level of Evidence: Therapeutic IV.

We report a case of free vascularized, second-toe metatarsophalangeal joint transfer in a 34-year-old manual laborer. The metatarsophalangeal joint of the left second toe was transferred to the metacarpophalangeal joint of the right ring finger in a reversed fashion. Fourteen years after surgery the transferred joint is painless and stable and flexion is 80 degrees with extension lag of 52 degrees without arthritic changes on radiographs.

An autopsy case of penetrating atheromatous ulcer of the ascending thoracic aorta is presented. The triplet imaging findings of mediastinal dilatation, interstitial pulmonary hemorrhage and pericardial effusion were diagnostic for penetrating atheromatous ulcer at the posterior wall of the ascending thoracic aorta. Immediate surgical treatment is indicated when these three findings emerge during examination.

Peroxisome proliferator-activated receptor (PPAR) gamma is a nuclear receptor known to regulate adipogenesis. Deletion of the PPARgamma gene in the mouse results in death by embryonic day 10.0 (E10.0) due to the failure of establishment of a labyrinth layer in the placenta, which suggests that PPARgamma is involved in trophoblast differentiation. To define PPARgamma function further in placental development, the expression and localization of the PPARgamma gene in the rat placenta was investigated. RT-PCR analysis shows the presence of PPARgamma mRNA in the placenta of day 11 of pregnancy (d11). The expression level is higher at d13 and then later decreased. Immunohistochemistry detects both PPARgamma and its putative intrinsic ligand, 15-deoxy-Delta(12,14)-prostaglandin J(2), in the trophoblast of layer I which lined the maternal sinus. Oral administration of troglitazone, an agonist of PPARgamma, to pregnant rats between d9 and d11 increases the expression level of PPARgamma in the placenta and reduces the mortality of the fetuses by half. These results suggest that PPARgamma is required not only for trophoblast differentiation but also trophoblast maturation to establish maternal-fetal transport.

Inducible nitric oxide (NO) synthase (iNOS) is believed to contribute to the pathogenesis of endotoxin-induced uveitis (EIU). In the present study, we investigated the inhibitory effects of N(G)-nitro-L-arginine methyl ester (L-NAME), a non-selective NOS inhibitor, and S,S'-1,4-phenylene-bis(1,2-ethanediyl)bis-isothiourea (PBITU), a potent and selective iNOS inhibitor, on intraocular NO production in EIU rabbits using an in vivo intraocular microdialysis technique. The flare level in the anterior chamber increased from 1h after the injection of 100 micro g/kg lipopolysaccharide (LPS), and continued to increase for 24h. Aqueous humor protein concentrations were significantly increased at 24h after LPS-injection. These changes were significantly reduced by L-NAME (10mg/kg) and PBITU (1mg/kg), but not by D-NAME (10mg/kg). The increase in NO(2)(-) and NO(3)(-) levels in the dialysate induced by LPS was significantly inhibited by L-NAME (10mg/kg) and PBITU (1mg/kg), but not by D-NAME (10mg/kg). These results suggest that activation of iNOS may play a key role in the development of EIU, and selective inhibitors of iNOS may have therapeutic applications in the treatment of EIU.

Cutaneous sensory nerves mediate inflammation and wound healing by releasing neuropeptides, such as substance P, which stimulates pro-inflammatory responses by keratinocytes, fibroblasts, and endothelial cells. The cell surface enzyme, neutral endopeptidase, degrades substance P, thereby regulating its biologic actions. We hypothesized that neutral endopeptidase enzymatic activity is increased in chronic wounds and skin from subjects with diabetes. We compared cutaneous neutral endopeptidase expression and enzymatic activity between normal controls and diabetic subjects with neuropathy and chronic wounds. Skin samples from subjects with diabetes were taken at the time of amputation for nonhealing ulcers. Skin taken from the ulcer margin, 1 cm from the ulcer (adjacent), and from the most proximal region of the amputated leg were studied. Skin biopsies from the leg of healthy control subjects were also studied. Neutral endopeptidase was localized by immunohistochemistry in all tissue sections. Neutral endopeptidase activity was measured using a fluorimetric assay. The median neutral endopeptidase activity of the ulcer margin was 1.21 x higher (p>0.2) than adjacent skin, 5.26 (p<0.001) than proximal skin, and 15.22 x higher (p<0.001) than control skin. Adjacent skin had a median neutral endopeptidase activity 4.34 x higher (p<0.001) than proximal skin and 12.58 x higher (p<0.001) than control skin. The median neutral endopeptidase activity of proximal skin was 2.90 x higher (p<0.001) than control skin. This elevated neutral endopeptidase activity in the skin and chronic ulcers of subjects with diabetes combined with peripheral neuropathy may contribute to deficient neuroinflammatory signaling and may impair wound healing in subjects with diabetes.

Peripheral steroid hormones act on brain tissues through intracellular receptor-mediated mechanisms to regulate several important brain neuronal functions. Therefore, the brain is considered to be a target site of steroid hormones. However, it is now established that the brain itself also synthesizes steroids de novo from cholesterol. The pioneering discovery of Baulieu and his colleagues, using mammals, and our studies with non-mammals have opened the door of a new research field. Such steroids synthesized in the brain are called neurosteroids. Because certain structures in vertebrate brains have the capacity to produce neurosteroids, identification of neurosteroidogenic cells in the brain is essential to understand the physiological role of neurosteroids in brain functions. Glial cells are generally accepted to be the major site for neurosteroid formation, but the concept of neurosteroidogenesis in brain neurons has up to now been uncertain. We recently demonstrated neuronal neurosteroidogenesis in the brain and indicated that the Purkinje cell, a typical cerebellar neuron, actively synthesizes several neurosteroids de novo from cholesterol in both mammals and non-mammals. Pregnenolone sulfate, one of neurosteroids synthesized in the Purkinje neuron, may contribute to some important events in the cerebellum by modulating neurotransmission. Progesterone, produced as a neurosteroid in this neuron only during neonatal life, may be involved in the promotion of neuronal and glial growth and neuronal synaptic contact in the cerebellum. More recently, biosynthesis and actions of neurosteroids in pyramidal neurons of the hippocampus were also demonstrated. These serve an excellent model for the study of physiological roles of neurosteroids in the brain, because both cerebellar Purkinje neurons and hippocampal neurons play an important role in memory and learning. This paper summarizes the advances made in our understanding of neurosteroids, produced in neurons, and their actions.

Objective/background: To identify the etiologic agent of rapidly progressive outer retinal necrosis (PORN) in a 32-year-old man with acquired immunodeficiency syndrome (AIDS), who had retinitis developed from cytomegalovirus (CMV). Multiple yellowish spots appeared in the deep retina without evidence of intraocular inflammation or retinal vasculitis, diagnosed clinically as PORN. Death occurred after failure of multiple organs., Design: Case report., Methods: Both globes were taken at autopsy, fixed in formalin, and examined histopathologically and immunohistochemically to identify causative agents in the retinal lesions., Main Outcome Measure: Immunohistochemistry., Results: All layers of the retina were severely damaged and contained focal calcification. Cytomegalic inclusion bodies were found in cells in the damaged retina of the right eye. Immunohistochemical studies for herpesviruses revealed the presence of CMV antigens in the right retina at the posterior pole and herpes simplex virus type 1 (HSV-1)-specific antigen in the periphery of both retinas. No varicella-zoster virus (VZV) antigen was detected in either retina., Conclusions: PORN has been described as a variant of necrotizing herpetic retinopathy, occurring particularly in patients with AIDS. Although the etiologic agent has been reported to be VZV, HSV-1 can be an etiologic agent.

Nitric oxide (NO) plays a role in controlling vascular tone and regulates the contractile properties of cardiac myocytes. Patients with heart failure exhibit high plasma levels of nitrite/nitrate (NOx), a stable metabolite of NO, and of cytokines such as tumor necrosis factor-alpha, a potent inducer of NO synthase. An increase in inducible NO synthase activity has been found in cardiac tissue from patients with dilated cardiomyopathy. These findings raise the possibility that local or systemic overproduction of NO induced by cytokines exerts a chronic negative inotropic effect on the myocardium and may have detrimental effects on systemic hemodynamics in patients with heart failure. Plasma levels of NG,NG-dimethylarginine (asymmetric dimethylarginine; ADMA), a circulating endogenous NO synthase inhibitor, were measured in control subjects and patients with valvular, hypertensive, or ischemic heart diseases or idiopathic cardiomyopathy. The plasma levels of NOx and ADMA were assessed by high performance liquid chromatography. The plasma levels of NOx and ADMA were significantly elevated in patients with heart failure. Both NOx and ADMA were positively correlated with New York Heart Association functional class. There was a significant inverse correlation between plasma NOx and ejection fraction, as estimated by echocardiography. A significant relationship between plasma NOx and ADMA was found only in patients with moderate to severe heart failure (r=0.41, p=0.01). Findings suggest a compensatory role of a circulating endogenous NO synthase inhibitor against induced NO synthase activity in patients with heart failure.

Parathyroid-hormone-related protein (PTHrP) is widely expressed not only in malignant tumors but also in both epithelial and nonepithelial cells of normal tissues. Secreted PTHrP is suspected to act as a paracrine or autocrine regulator. However, little is known about its secretory pathway. To cast light on this question, we studied the intracytoplasmic distribution of parathyroid hormone (PTH) and PTHrP immunohistochemically and immunoelectron microscopically in 10 surgically resected parathyroid adenomas. Double immunostaining was performed using anti-PTH antibody and a newly established anti-PTHrP antibody to reveal the relationship between their two distributions. Additional examination by cell immunoblot assay was performed to determine whether both PTH and PTHrP are secreted simultaneously. Both PTH and PTHrP were actually secreted from individual parathyroid cells simultaneously on cell immunoblot assay. Immunohistochemically, there were two different types of adenoma cells, i.e., one positive only for PTH and the other positive for both PTH and PTHrP. PTH was distributed linearly or fine granularly along the cytoplasmic membrane, whereas PTHrP was distributed diffusely or coarse granularly in the cytoplasm. The intracytoplasmic distributions of PTH and PTHrP often overlapped. Immunoelectron microscopical examination demonstrated that PTHrP co-localized with PTH in the same secretory granules. The results clearly demonstrated that PTHrP can be co-secreted with PTH via a regulated pathway using secretory granules.

Sugi basic protein (SBP), a major allergen of Japanese cedar (Cryptomeria japonica) pollen, conjugated to pullulan (alpha-1,4'-, alpha-1,6'-glucan) reportedly suppresses IgE anti-SBP antibody production and enhances IgG anti-SBP antibody production in mice. We analyzed cytokine production by SBP-specific T cells after stimulation with an SBP-pullulan conjugate (SBP-P), native SBP, or a mixture of SBP and pullulan. When SBP-specific T cell lines were stimulated with the SBP-P conjugate in the presence of antigen-presenting cells (APC), the production of IFN-gamma, IL-4, IL-5, and IL-10 decreased compared with the cytokine levels produced by SBP-stimulated T cells. However, when these T cells were repeatedly stimulated with the SBP-P conjugate, the production of IFN-gamma increased progressively, while that of IL-4, IL-5, and IL-10 remained decreased compared with the T cells that were repeatedly stimulated with native SBP. Stimulation of the T cells with the mixture of SBP and pullulan showed little difference in the cytokine production profile from that observed after stimulation with native SBP alone. Interestingly, when the T cell lines stimulated repeatedly with SBP-P were subsequently stimulated with native SBP, a further increase in IFN-gamma production was observed, while IL-10 production decreased. Inhibition of IL-4 production was also observed when SBP-specific Th2 clones were stimulated with SBP-P. These results indicate that stimulation of T cells with SBP-P up-regulates Th1 cytokine production, while down-regulating that of Th2. It is, therefore, conceivable that immunotherapeutic treatment with the SBP-P conjugate rather than with conventional SBP solutions is preferable for improving Japanese cedar pollen allergy.

The relationship between parathyroid hormone-related protein (PTHrP) release from parathyroid cells and extracellular calcium ion concentration was investigated in three cases of parathyroid hyperplasia secondary to chronic renal failure and in four cases of parathyroid adenoma. Amounts of PTHrP released from individual parathyroid cells dispersed from surgical specimens were estimated by cell immunoblot assay. Parathyroid cells from both hyperplasias and adenomas showed significant suppression in the release of PTHrP with increase in extracellular calcium ions, but the amounts of PTHrP released from adenoma cells were significantly larger than from hyperplasia cells. The maximal value for PTHrP released within 120 minutes from adenoma cells was 2.91 +/- 2.11 x 10(-2) fmol/cell ([Ca2+], 0.4 mmol/L), and the minimal value was 1.32 +/- 0.35 x 10(-2) fmol/cell ([Ca2+], 2.0 mmol/L). On the other hand, the maximal value for PTHrP released from hyperplasia cells was 1.79 +/- 1.56 x 10(-2) fmol/cell ([Ca2+], 0.4 mmol/L), and the minimal value was 0.32 +/- 0.19 x 10(-2) fmol/cell ([Ca2+], 2.0 mmol/L). These results demonstrate actual release of PTHrP from abnormal parathyroid tissues into the extracellular space with the response to extracellular calcium ions depending on the cell status. Given the lack of definite histological criteria to differentiate between hyperplasias and adenomas in the parathyroid gland, the presently demonstrated significant difference in the ability to release PTHrP is important in pointing to parathyroid hyperplasia secondary to chronic renal failure as a distinct pathological entity separate from parathyroid adenoma.

We studied the effect of flexor tendon sheath closure on flexor tendon function after delayed primary flexor tendon suture in white leghorn chickens. The tendon suture was carried out after a tendon laceration. In the left foot the tendon sheath was closed after tendon suture, and in the right the sheath was excised over the tendon suture. Tendon gliding, joint motion, the fate of the closed sheath, peritendinous adhesions, and tendon healing were studied. The sutured sheath disappeared after suturing and was associated with poor tendon healing. Sheath closure did not improve flexor tendon function in a delayed primary repair.

Dorsal and circumferential flexor sheaths were reconstructed by autogenous sheath graft in flexor tendon repair in 28 white leghorn chickens. The effect of the sheath reconstructions was evaluated by use of an experimental model of tendon transection associated with bony injury, superficial tendon excision, and 3 weeks of immobilization. Six weeks after reconstruction, the gliding excursion was assessed and adhesion formation, tendon healing, and survival of the grafted sheath were evaluated both macroscopically and histologically. Dorsal and circumferential sheath grafts resulted in tendon gliding that was significantly better than the group with sheath defect not reconstructed. The adhesion formation was apparently less severe, and tendon healing was better in the groups with sheath reconstruction than in the group without reconstruction. The group with dorsal sheath reconstruction did not differ significantly from that with circumferential reconstruction in gliding excursion and adhesion formation. This study demonstrates that separation of the injured tendons from the bony surface by autogenous sheath grafts may be beneficial to tendon gliding and for reducing adhesion formation.

We report a case of multifocal neurilemomas originating from different nerves in the same upper extremity. In the excision of all seven tumors care was taken not to damage the nerves involved. The common features of multifocal neurilemomas that originated from different nerves of the same upper extremity are outlined and the consequent risk of potential nerve damage during the operation is discussed. We recommend intracapsular excisions of multifocal neurilemomas.

Rat-mouse hybridoma cells producing anti-mouse IgE antibodies were intraperitoneally or subcutaneously inoculated into newborn or suckling hamsters receiving rabbit anti-hamster thymocyte globulin from the day of birth twice a week for at least 3 weeks. The hybridoma cells were found to grow in the abdominal cavity of the hamsters as ascites tumor or in subcutaneous tissue as solid tumor without loss of antibody-secreting activities. For the production of ascites, 2-week-old hamsters were preferable to newborn hamsters. In 3-week-old hamsters, the hybridoma cells could scarcely survive. The antibody titers of the ascites were determined to be 10(5)-10(6) in the ELISA and in the ability to neutralize the skin-sensitizing capacity of mouse IgE antibodies. The rat monoclonal antibodies were easily separated from ascites, serum or cell culture supernatant with affinity chromatography using Affigel protein A-Sepharose and anti-hamster IgG-Sepharose columns. The described method could be efficiently applicable for the proliferation of other hybridomas, such as human-human, human-mouse or hamster-mouse, etc.

The sensibility of 25 fingers reconstructed with three different types of free sensory flaps from the toe was evaluated. The relationships between sensory restoration and the patient's age, type of sensory flap, selection of sutured nerves, and circulatory complications in the flap were analyzed. Wrap-around flaps were used in 5 fingers, hemipulp flaps in 11, and toe-to-finger transfers in 9. The follow-up period ranged from 16 to 79 months. Restoration of sensibility in the flap was closely related to the patient's age and vascular insufficiency of the flap. Therefore, establishing and maintaining and adequate circulation in the flap at operation is essential for good sensory restoration.

Hind legs of dogs were amputated at the middle of the thigh and preserved in three different conditions: in ice water, in a refrigerator, and at room temperature. After 6 or 12 hours of ischemia, recirculation was established. The survival rate of the animals was observed and measurement of limb edema, potassium, pH, and lactate in the blood was performed to study the effects of hypothermia on prevention of "replantation toxemia." Cooling of the amputated limb was effective for prevention of toxemia, and the cooling effect was greater in ice water than in a refrigerator. However, when cooled in ice water, some animals died due to toxemia when the time of ischemia was prolonged to 12 hours. In the dead animals, a close relationship was observed between the developement of toxemia and metabolic acidosis due to the increase in lactate.

Of 32 patients with Kienböck's disease, 19 had lunate silicone implant replacements, and 13 had coiled palmaris longus tendon replacement. The carpal height was measured and osteoarthritic changes were noted in plain x-ray films before and after the operation. Follow-up periods ranged from 3 to 11 years, with an average of 6 years 4 months. In patients in the mild stages of the preoperative carpal collapse, both excisional arthroplasties had good clinical results. The x-ray films showed that a silicone implant played a more important role in preventing further carpal collapse than a palmaris longus tendon replacement in patients in the early carpal collapse stage. However, the clinical results of the silicone implant replacement were unsatisfactory because of the postoperative progression of osteoarthritic changes or subluxation of the prosthesis in the advanced stages of carpal collapse. The palmaris longus tendon replacement gave satisfactory results in four of seven patients with advanced carpal collapse. These results suggest that silicone implant replacement is indicated primarily in patients with the early stages of carpal collapse, and replacement of the palmaris longus tendon is rarely recommended in either the early or late stages of carpal collapse.

The immunologic antigenicity of tendons after various treatments was investigated by means of the complement-dependent cytotoxicity test in two strains of inbred rats. Allogeneic tendons treated by various methods were grafted into rats, and the survival of the grafted tendons was observed macro- and microscopically. Paraformaldehyde-treated allogeneic tendons showed the least immunologic response. The remaining treated allogeneic tendons showed moderate to marked immunologic responses. These in vivo results correlated with those obtained in previous in vitro experiments.