Study on Distribution of Four Pseudomonas Species in Living Environment Using Multiplex PCR

Purpose: The genus Pseudomonas is a ubiquitous microorganism frequently detected from immunocompromised patients. The inherent resistance to numerous antimicrobial agents contributes to the opportunistic character of this pathogen exhaustive monitoring of this pathogen is considered of critical importance to public health organizations. The reliable identification method able to distinguish genetic close Pseudomonas species is needed, because these organisms are difficult to differentiate by phenotypic or biochemical methods. The purpose of the present study was to design species-specific primers in order to identify and detect four Pseudomonas species which are frequently detected from the human oral cavities, and to investigate the distribution of these organisms in the living environment using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the rpoD gene of four Pseudomonas species. Swab samples were collected from fifty washstands, and the distribution of Pseudomonas species was investigated using a conventional PCR at genus level and a multiplex PCR at species level. Results: Multiplex PCR method developed in this study was able to distinguish four Pseudomonas species clearly. The genus Pseudomonas was detected from all samples (100%), whereas P. putida, P, aeruginosa, P. stutzeri and P. fluorescens were detected at 44%, 8%, 4% and 2% in fifty swab samples, respectively. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and works without requiring DNA extraction. It was indicated that washstands were the uninhabitable environment for P. putida, P, aeruginosa, P. stutzeri and P. fluorescens.

Quantification of Ride Comfort Using Musculoskeletal Mathematical Model Considering Vehicle Behavior

This research aims to quantify driver ride comfort due to changes in damper characteristics between comfort mode and sport mode,considering the vehicle’s inertial behavior.The comfort of riding in an automobile has been evaluated in recent years on the basis of a subjective sensory evaluation given by the driver.However,reflecting driving sensations in design work to improve ride comfort is abstract in nature and difficult to express theoretically.Therefore,we evaluated the human body’s effects while driving scientifically by quantifying the driver’s behavior while operating the steering wheel and the behavior of the automobile while in motion using physical quantities.To this end,we collected driver and vehicle data using amotion capture system and vehicle CAN and IMU sensors.We also constructed a three-dimensional musculoskeletal mathematical model to simulate driver movements and calculate the power and amount of energy per unit of time used for driving the joints and muscles of the human body.Here,we used comfort mode and sport mode to compare damper characteristics in terms of hardness.In comfort mode,damper characteristics are soft and steering stability is mild,but vibration from the road is not easily transmitted to the driver making for a lighter load on the driver.In sport mode,on the other hand,damper characteristics are hard and steering stability is comparatively better.Still,vibration from the road is easily transmitted to the driver,whichmakes it easy for a load to be placed on the driver.As a result of this comparison,it was found that a load was most likely to be applied to the driver’s neck.This result in relation to the neck joint can therefore be treated as an objective measure for quantifying ride comfort.

Silicone oil injection and removal in 27-gauge vitreous surgery

Dear Editor,The advent of 23-gauge(G),25-G,and 27-G vitrectomy systems[1]since the beginning of the 21stcentury has allowed vitrectomy via a smaller incision compared to the 20-G system;hence the name"microincision vitrectomy surgery".Microincision vitrectomy surgery has gainedpopularity among vitreoretinal surgeons.

Histopathological and Immunohistochemical Study of Peri-Implant Epithelium

Background: Human beta-defensins (HBDs), epithelial-derived antimicrobial peptides, contribute to gingival health and periodontal disease. Particularly, the upregulation of HBD-3 expression in periodontal tissues is described to contribute to periodontal tissue regeneration. Specifically, the upregulation of HBD-3 expression in periodontal tissues is described to contribute to periodontal tissue regeneration. But there has been insufficient investigation of the inner epithelium in peri-implantitis, especially the HBD expression patterns as protection against oral infection. Objectives: The purpose of this study was to investigate the histopathological characteristics of the inner implant epithelium in peri-implantitis using histopathological and immunohistochemical methods and to compare it with the inner marginal epithelium in periodontitis. Materials and Methods: The biopsied cases consisted of 10 peri-implantitis, 11 periodontitis, and 10 controls. HE observation, measurement of the thickness of the inner epithelium, and immunohistochemical analysis for Ki-67 and HBD-3 were conducted. Results: Concerning the thickness of the inner epithelium, it was significantly higher in periodontitis (156.2 [138.0, 186.4] μm) than in peri-implantitis and control, 70.7 [67.5, 97.5] μm and 80.7 [76.6, 89.4] μm, respectively (p Conclusion: Decreased production of HBD-3 in keratinocytes composing the inner implant epithelium may be one of the factors affecting tissue repair in peri-implantitis.

Investigation of Bacteria Species Most Involved in Peri-Implantitis

Purpose: Currently, bacteriological examinations of implant treatments target periodontopathic bacteria such as red complex bacteria, including Porphyromonas gingivalis, and detect them qualitatively or quantitatively. However, it seems that those examinations do not reflect the peri-implant tissue conditions precisely, because periodontopathic bacteria are also frequently detected from healthy peri-implant sites. The purpose of the present study was to investigate bacteria species most involved in peri-implantitis using a PCR method. Methods: Polymerase chain reaction (PCR) primers in this study were designed based on partial sequences of 16S rDNA of bacteria species involved in peri-implantitis that were described in numerous previous studies. Peri-implant sulcus fluid (PISF) samples were collected from thirty periodontally healthy patients with implants (HI) and thirty patients with peri-implantitis (PI). Each detection frequency of bacteria species in PISFs of both groups was investigated using a PCR method, and was compared using Fisher’s exact test. Results: In PI group, detection frequencies of Corynebacterium durum, Fretibacterium fastidiosum and Slackia exigua were significantly higher than those of HI group (p P. gingivalis and Tannerella forsythia belonging to red complex were frequently detected in the PISF samples of HI group (p > 0.05). Conclusion: It was suggested that monitoring C. durum and F. fastidiosum levels in PISF samples was useful as a clinical indicator for the evaluation of peri-implant tissue conditions.

Isolation and Identification Methods for Oral Klebsiella pneumoniae Involved in Onset of Inflammatory Bowel Disease

Purpose: Recently, it was reported that Klebsiella pneumoniae is related to the onset of inflammatory bowel disease including the Crohn disease. It was frequently reported that K. pneumoniae was detected in human oral cavities. Regrettably, it currently remains unclear whether K. pneumoniae is part of the normal oral flora. The aim of this study was to establish the isolation and identification methods for K. pneumoniae from human oral cavities, and investigate its transmission pattern. Methods: A selective medium, OKPSM, for the isolation of K. pneumoniae from oral cavities was developed in this study. Also, PCR primer for the identification and detection at subspecies level of K. pneumoniae was designed. Results: OKPSM and PCR method using the primers designed in this study were useful for the isolation and identification of K. pneumoniae from human oral cavities. K. pneumoniae subsp. pneumoniae was detected at 10.0% in 30 saliva samples. On the other hand, K. pneumoniae subsp. ozaenae and K. pneumoniae subsp. rhinoscleromatis were detected from no sample. Moreover, K. pneumoniae subsp. pneumoniae isolates from same subject at 0 month and after 3 months showed same genotypes on AP-PCR using OPA-07 primer. Conclusion: These results indicated that human oral cavities were not suitable for the habitat of K. pneumoniae.

Study on Distribution of Acinetobacter baumannii Complex in Dental Hospital Using Multiplex PCR

Purpose: In recent years, multidrug-resistant Acinetobacter baumannii has appeared and caused outbreaks of hospital infections all over the world. Close monitoring of this pathogen and other A. baumanii complex species is considered of critical importance to public health organizations. The reliable identification method able to distinguish A. baumannii from genetically close Acinetobacter species is needed, because these species are unable to be differentiated by phenotypic or biochemical methods. The purpose of the present study was to design species-specific primers in order to identify and detect A. baumanii complex species, and Acinetobacter lwoffii which is frequently detected from the human specimens, and to investigate the distribution of these organisms in dental hospital using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the 16S ribosomal RNA (16S rRNA) gene and DNA gyrase subunit B (gyrB) of each species of A. baumanii complex species. Swab samples were collected from ten dental spittoon units in dental hospital, and the distribution of A. baumanii complex species was investigated using a multiplex PCR. Results: These primers were able to distinguish each species of A. baumanii complex species clearly. A. baumanii and A. calcoaceticus were detected at 20.0% and 10.0% in ten swab samples, respectively. On the other hand, A. nosocomialis, A. lowffii, and A. pittii were detected from no sample. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and worked without requiring DNA extraction.

Comparative Cytological and Histopathological Study of Peri-Implantitis and Periodontitis

Peri-implant diseases, such as peri-implant mucositis and peri-implantitis, pose significant challenges to the long-term prognosis of dental implants. This study aimed to comprehensively compare peri-implantitis with periodontitis from cytological and histopathological perspectives, shedding light on the morphological characteristics associated with peri-implantitis. Thirteen patients, including six with peri-implantitis and seven with periodontitis, were included in the study. Cytological examination of affected gingival mucosa revealed distinct differences between the two conditions. Peri-implantitis exhibited an inflammatory background predominantly composed of neutrophils with lobulated nuclei, accompanied by stratified squamous epithelial cells showing signs of keratinization. In contrast, periodontitis showed a similar neutrophilic inflammatory background but with non-keratinized epithelial cells. Histopathological examination further confirmed these differences, with peri-implantitis showing keratinized epithelium in the inner epithelial layer. This histological finding aligns with the notion that peri-implantitis has a distinct mucosal profile compared to periodontitis. Additionally, cytological analysis revealed that peri-implantitis had a lower occurrence rate of Light green-positive cells, indicating a tendency toward keratinization. This finding suggests that the presence of keratinized mucosa might be associated with peri-implant health, although further research is needed to clarify this relationship. Overall, this study demonstrates the potential of cytological examination and Papanicolaou staining for assessing mucosal inflammatory conditions and distinguishing between keratinized and non-keratinized cells. These findings underscore the utility of oral mucosal smears as a valuable tool for diagnosing peri-implantitis and enhancing our understanding of its pathogenesis.

Mesenchymal stem cell-derived apoptotic bodies alleviate alveolar bone destruction by regulating osteoclast differentiation and function

Periodontitis is caused by overactive osteoclast activity that results in the loss of periodontal supporting tissue and mesenchymal stem cells(MSCs)are essential for periodontal regeneration.However,the hypoxic periodontal microenvironment during periodontitis induces the apoptosis of MSCs.Apoptotic bodies(ABs)are the major product of apoptotic cells and have been attracting increased attention as potential mediators for periodontitis treatment,thus we investigated the effects of ABs derived from MSCs on periodontitis.MSCs were derived from bone marrows of mice and were cultured under hypoxic conditions for 72 h,after which ABs were isolated from the culture supernatant using a multi-filtration system.The results demonstrate that ABs derived from MSCs inhibited osteoclast differentiation and alveolar bone resorption.miRNA array analysis showed that miR-223-3p is highly enriched in those ABs and is critical for their therapeutic effects.Targetscan and luciferase activity results confirmed that Itgb1 is targeted by miR-223-3p,which interferes with the function of osteoclasts.Additionally,DC-STAMP is a key regulator that mediates membrane infusion.ABs and pre-osteoclasts expressed high levels of DC-STAMP on their membranes,which mediates the engulfment of ABs by pre-osteoclasts.ABs with knock-down of DC-STAMP failed to be engulfed by pre-osteoclasts.Collectively,MSC-derived ABs are targeted to be engulfed by pre-osteoclasts via DC-STAMP,which rescued alveolar bone loss by transferring miR-223-3p to osteoclasts,which in turn led to the attenuation of their differentiation and bone resorption.These results suggest that MSC-derived ABs are promising therapeutic agents for the treatment of periodontitis.

One Step Multiplex PCR for Identification of Candida haemulonii complex and Candia auris

Purpose: Recently, Candida haemulonii complex (Candida haemulonii, Candida duobushaemulonii and Candida haemulonii var. vulnera) and two genetically close species (Candida pseudohaemulonii and Candida auris) have emerged as an opportunistic fungal pathogen associated with various infectious diseases of humans, and most of those isolates have displayed antifungal resistance. Because it is difficult to differentiate these microorganisms by a current technique, unfortunately, it is important to establish a method for identifying them accurately. The purpose of the present study was to design species-specific primers in order to identify and detect C. auris, C. pseudohaemulonii, and C. haemulonii complex, i.e. , C. haemulonii, C. duobushaemulonii and C. haemulonii var. vulnera using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the 26S rRNA, 18S rRNA, and RPB1 genes and ITS region of five Candida species. Results: The multiplex PCR method developed in this study was able to distinguish five Candida species clearly. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and works without requiring DNA extraction.

岩石剪切面纳米粒子层的近期研究进展

20世纪90年代,在各类岩石剪切面中相继发现纳米级和微米级超微粒子结构,近期结合力学、物理学成因研究又有新的进展.在超微观上,通过国内外的实例实测,从岩石剪切面纳米粒子层赋存展布的普遍性,结构特征的层次性,成因机制的非线性和功能效应的多样性四个方面,进行简要阐述,萃取更多信息.进而初步揭示了剪切摩擦黏性发育与纳米粒子形成间的共生关系,和剪切运动中岩石的分离、分凝和分层作用首先由纳米粒子层的滑移启动开始的.并分析了岩石塑性流变实质上是超微粒子(纳米-微米级磨粒)的运动,以及岩石剪切面纳米粒子层一些特殊的物理和化学地质作用等.诚然,纳米粒子层蕴涵的信息是丰富的,该项研究还处于起步开拓阶段,仅试图为构造地质学和相关学科领域提出新的理念和新的探讨平台.

小型激光天文动力学空间计划概念

小型激光天文动力学空间计划是 :使用在太阳轨道上无拖曳航天器和地面站以激光干涉和脉冲测距的方法 ,精确地探讨天文动力学 ,检测相对论与时空基本定律 ,改进探测引力波的灵敏度以及更准确地测定太阳、行星和小行星的参数。 1 969年开始的月球激光 (反射 )测距 ,对地球物理、参考坐标的选定、相对论的检验均有重要的贡献。 3 0年来 ,激光技术的长足进步 ,使现在正是适合于开始进行研究空间有源 (主动 )测距和光波空间通讯的时候。激光天文动力学的兴起是必然的趋势 ,其精确度将比现在提高 3到 6个数量级 ,将是天文动力学革命性的发展。小型激光天文动力学空间计划可以起到带头作用。它的关键技术有三 ,即 :弱光锁相、极精确无拖曳航天和高衰减日冕仪。弱光锁相已有长足的进步。对高衰减日冕仪的研究 ,也有了初步的方案。LISA空间计划将于 2 0 0 6年 8月发射SMART -2 ,研究测试极精确无拖曳航天。小型激光天文动力学空间计划的关键技术已日趋成熟。在第一届国际激光天文动力学研讨会 ( 2 0 0 1 ,9.1 3 -2 3 )中介绍了各相关学科背景及前沿研究 ,讨论了激光天文动力学空间计划科学目标及相关技术 ,并召开了两次小型激光天文动力学空间计划预研究筹备会 ,建立了和欧洲的合作关系。会后着手进行此项对基础?

Apoptosis and non-alcoholic fatty liver diseases

The number of patients with nonalcoholic fatty liver diseases(NAFLD) including nonalcoholic steatohepatitis(NASH), has been increasing. NASH causes cirrhosis and hepatocellular carcinoma(HCC) and is one of the most serious health problems in the world. The mechanism through which NASH progresses is still largely unknown. Activation of caspases, Bcl-2 family proteins, and c-Jun N-terminal kinase-induced hepatocyte apoptosis plays a role in the activation of NAFLD/NASH. Apoptotic hepatocytes stimulate immune cells and hepatic stellate cells toward the progression of fibrosis in the liver through the production of inflammasomes and cytokines. Abnormalities in glucose and lipid metabolism as well as microbiota accelerate these processes. The production of reactive oxygen species, oxidative stress, and endoplasmic reticulum stress is also involved. Cell death, including apoptosis, seems very important in the progression of NAFLD and NASH. Recently, inhibitors of apoptosis have been developed as drugs for the treatment of NASH and may prevent cirrhosis and HCC. Increased hepatocyte apoptosis may distinguish NASH from NAFLD, and the improvement of apoptosis could play a role in controlling the development of NASH. In this review, the association between apoptosis and NAFLD/NASH are discussed. This review could provide their knowledge, which plays a role in seeing the patients with NAFLD/NASH in daily clinical practice.

Improving recognition of hepatic perivascular epithelioid cell tumor:Case report and literature review

A 58-year-old man presented with the chief complaint of abdominal bloating and was incidentally found to have a liver tumor.As diagnostic imaging studies could not rule out malignancy,the patient underwent partial resection of segment 3 of the liver.The lesion pathologically showed eosinophilic proliferation,in addition to immunohistochemical positivity for human melanoma black 45 and Melan-A,thereby leading to the diagnosis of a hepatic perivascular epithelioid cell tumor(PEComa).A PEComa arising from the liver is relatively rare.Moreover,the name ‘PEComa' has not yet been widely recognized,and the same disease entity has been called epithelioid angiomyolipoma(EAML),further diminishing the recognition of PEComa.In addition,PEComa imaging findings mimic those of malignant liver tumors,and clinically,this tumor tends to enlarge.Therefore,a PEComa is difficult to diagnose.We conducted a systematic review of PEComa and EAML cases and discuss the results,including findings useful for differentiating perivascular epithelioid cell tumors from malignant liver tumors.

Development of Helicobacter pylori treatment: How do we manage antimicrobial resistance?

Helicobacter pylori(H. pylori) antimicrobial resistance is an urgent, global issue. In2017, the World Health Organization designated clarithromycin-resistant H.pylori as a high priority bacterium for antibiotic research and development. In addition to clarithromycin, resistance to metronidazole and fluoroquinolones has also increased worldwide. Recent international guidelines for management of H.pylori infection recommend bismuth or non-bismuth quadruple therapy for 14 d as a first-line treatment for H. pylori in areas of high clarithromycin and/or metronidazole resistance. Although these treatment regimens provide acceptable H. pylori eradication rates, the regimens used should not contribute to future resistance of H. pylori to antimicrobials. Moreover, these regimens can promote resistance, due to prolonged therapy with multiple antibiotics. A new strategy that can eradicate H. pylori as well as reduce the antibiotics used is required to prevent future antimicrobial resistance in H. pylori. Dual-therapy with vonoprazan and amoxicillin could be a breakthrough for H. pylori eradication in an era of growing antimicrobial resistance. This regimen may provide a satisfactory eradication rate of H. pylori and also minimize antimicrobial resistance due to single antibiotic use and the strong inhibitory effect of vonoprazan on gastric acid secretion.

Anti-inflammatory activity of probiotic Bifidobacterium: Enhancement of IL-10 production in peripheral blood mononuclear cells from ulcerative colitis patients and inhibition of IL-8 secretion in HT-29 cells

AIM: To determine the anti-inflammatory activity of probiotic Bifidobacteria in Bifidobacteria-fermented milk (BFM) which is effective against active ulcerative colitis (UC) and exacerbations of UC, and to explore the immunoregulatory mechanisms. METHODS: Peripheral blood mononuclear cells (PBMNC) from UC patients or HT-29 cells were co-cultured with heat-killed probiotic bacteria or culture supernatant of Bifidobacterium breve strain Yakult (BbrY) or Bifidobacterium bifidum strain Yakult (BbiY) to estimate the amount of IL-10 or IL-8 secreted. RESULTS: Both strains of probiotic Bifidobacteria contained in the BFM induced IL-10 production in PBMNC from UC patients, though BbrY was more effective than BbiY. Conditioned medium (CM) and DNA of both strains inhibited IL-8 secretion in HT-29 cells stimulated with TNF-α, whereas no such effect was observed with heat- killed bacteria. The inhibitory effect of CM derived from BbiY was greater than that of CM derived from BbrY. DNAs of the two strains had a comparable inhibitory activity against the secretion of IL-8. CM of BbiY induced a repression of IL-8 gene expression with a higher expression of IκB-ζ mRNA 4 h after culture of HT-29 cells compared to that in the absence of CM.CONCLUSION: Probiotic Bifidobacterium strains in BFM enhance IL-10 production in PBMNC and inhibit IL-8 secretion in intestinal epithelial cells, suggesting that BFM has anti-inflammatory effects against ulcerative colitis.

Pericardiocentesis with cisplatin for malignant pericardial effusion and tamponade

AIM:To evaluate the role and outcome of pericardiocentesis with intrapericardial cisplatin instillation for malignant pericardial effusion resulting from esophageal cancer. METHODS:We retrospectively studied 7 patients who underwent pericardiocentesis with intrapericardial cisplatin instillation for malignant pericardial effusion resulting from esophageal cancer.After pericardiocentesis,we performed catheterization of the pericardial space under ultrasonogram guidance.Malignant etiology of the pericardial fluid was confirmed by cytological examination.Subsequently,cisplatin(10 mg in 20 mL normal saline) was instilled into the pericardial space. RESULTS:The mean total volume of the aspirated effusion fluid was 782±264 mL(range,400-1200 mL) . The drainage catheter was successfully removed in all patients,and the mean duration of pericardial drainagewas 7.7±2.7 d(range,5-13 d) .No fluid reaccumulation was observed.Mean survival time was 120±71 d(range,68-268 d) . CONCLUSION:Pericardiocentesis along with catheter drainage appears to be a safe and effective for pericardial malignant effusion and tamponade,and cisplatin instillation prevents recurrence.

Scoring system for prediction of metastatic spine tumor prognosis

Assessing the prognosis before treatment for metastatic spine tumor is extremely important in therapy selection.Therefore,we review some prognostic scoring systems and their outcomes.Articles with combinations of two keywords among"metastatic spine tumor"and"prognosis","score","scoring system","predicting",or"life expectancy"were searched for in Pub Med.As a result,236 articles were extracted.Those referring to representative scoring systems about predicting the survival of patients with metastatic spine tumors were used.The significance and limits of these scoring systems,and the future perspectives were described.Tokuhashi score,Tomita score,Baur score,Linden score,Rades score,and Katagiri score were introduced.They are all scoring systems prepared by combining factors that affect prognosis.The primary site of cancer and visceral metastasis were common factors in all of these scoring systems.Other factors selected to influence the prognosis varied.They were useful to roughly predict thesurvival period,such as,"more than one year or not"or"more than six months or not".In particular,they were utilized for decision-making about operative indications and avoidance of excessive medical treatment.Because the function depended on the survival period in the patients with metastatic spine tumor,it was also utilized in assessing functional prognosis.However,no scoring system had more than 90%consistency between the predicted and actual survival periods.Future perspectives should adopt more oncological viewpoints with adjustment of the process of treatment for metastatic spine tumor.

Adenocarcinoma arising from heterotopic pancreas at the third portion of the duodenum

A 62-year-old Japanese man presented to our hospital with a history of weight loss of 6 kg in 4 mo. Imaging examinations revealed a tumor located on the third portion of the duodenum with stenosis. We suspected duodenal carcinoma and performed pancreas-preserving segmental duodenectomy. Adenocarcinoma arising from a heterotopic pancreas at the third portion of the duodenum was finally diagnosed by immunohistochemical staining. Malignant transformation in the duodenum arising from a heterotopic pancreas is extremely rare; to our knowledge, only 13 cases have been reported worldwide, including the present case. The most common location of malignancy is the proximal duodenum at the first and descending portion. Herein, we describe the first case of adenocarcinoma arising from a heterotopic pancreas, which was located in the third portion of the duodenum, with a review of the literature.