Oral human cytomegalovirus prevalence and its relationships with periodontitis and Porphyromonas gingivalis in Japanese adults: a cross-sectional study

Objective: This study aimed to clarify the association between oral human cytomegalovirus (HCMV) and periodontitis in Japanese adults. Methodology: In total, 190 patients (75 men and 115 women; mean age, 70.2 years) who visited Hiroshima University Hospital between March 2018 and May 2020 were included. Oral rinse samples were taken to examine the presence of HCMV DNA using real-time polymerase chain reaction (PCR). P. gingivalis was detected by semi-quantitative PCR analysis. Results: HCMV DNA was present in nine of 190 patients (4.7%). There were significant associations between HCMV presence and the presence of ≥4-mm-deep periodontal pockets with bleeding on probing (BOP) (P<0.01) and ≥6-mm-deep periodontal pockets with BOP (P=0.01). However, no significant relationship was observed between HCMV presence and periodontal epithelial surface area scores. Logistic regression analysis revealed that the presence of ≥4-mm-deep periodontal pockets with BOP was significantly associated with HCMV (odds ratio, 14.4; P=0.01). Propensity score matching was performed between patients presenting ≥4-mm-deep periodontal pockets with BOP (i.e., active periodontitis) and patients without ≥4-mm-deep periodontal pockets with BOP; 62 matched pairs were generated. Patients who had ≥4-mm-deep periodontal pockets with BOP showed a higher rate of HCMV presence (9.7%) than those who lacked ≥4-mm-deep periodontal pockets with BOP (0.0%). There was a significant relationship between HCMV presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). A significant relationship was found between HCMV/P. gingivalis DNA presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). Conclusions: Coinfection of oral HCMV and P. gingivalis was significantly associated with active periodontitis. Moreover, interactions between oral HCMV and P. gingivalis may be related to the severity of periodontal disease

再利用を伴う日本での家庭浴槽水中の無機成分変化

For bathing, water is heated to warm the body, and is used to remove cleaning agents from the skin. Usually this water does not enter the body, but there have been cases where people have caught infectious diseases through inhalation of aerosols during bathing. In this study, as a model case of circulating bath, we examined the change in hygiene and chemical composition of the bathtub water in various conditions. To investigate the potential for bacterial growth in domestic bath water in Japan, changes in the concentrations of metal and non-metal inorganic compounds were studied as the number of days the water was re-used for increased. With re-use of bath water over 1 week, the concentrations of most of the compounds increased. However, the concentration of phosphorus decreased, and the concentrations of manganese and titanium showed both increases and decreases. Increasing concentrations could be attributed to sweat secreted from the skin of the bather, compounds eluted from the water heater as the bath water was re-heated, and dissolution of added bath salts. Consumption of inorganic compounds by bacteria and precipitation of metal hydr oxides could contribute to the decreasing concentrations. Common bacteria and coliform bacteria were measured simultaneously, and only the coliform bacteria increased in the water on the second day the water was used for bathing. Bacteria probably grew in the pipes between the water heater and the bat htub.入浴は，身体を温めたり，石鹸などの成分を皮膚から取り除くために行われる。通常，浴槽水は体内に入ることはないが，入浴中のエアロゾルの経気道的吸入により，感染症を引き起こすことが知られている。本研究では，循環型風呂のモデルとして，衛生学的に浴槽水の成分が様々な使用条件でどう変化するかを調べた。日本における家庭用浴槽水中での微生物繁殖の可能性を調べるため，金属，非金属無機化合物の日ごとの濃度変化を追跡した。浴槽水の再利用を1 週間行ったところ，多くの化合物濃度は増加した。リンのみが減少し，マンガンやチタンは増加と減少の両方が観察された。これらの変化において，増加は入浴者の皮膚から排泄された汗，風呂釜からの溶出，入浴剤からの混入が考えられた。減少は浴槽水中細菌による消費と金属水酸化物の沈殿析出が考えられた。同時に一般細菌や大腸菌群についても測定したが，大腸菌は入浴2 日目に増加し，風呂釜中に生育場所がある可能性が示唆された

A Quantitative Immunochromatography Assay of Whole Blood Samples for Antigen-specific IgE—A New Method for Point of Care Testing for Allergens—

ABSTRACTBackgroundThe development of an inexpensive point-of-care testing system for antigen-specific IgE is greatly needed. We, therefore, tried to develop a quantitative enzyme immunochromatography assay system for antigen-specific IgE in fresh whole blood.MethodsWhole blood sample was mixed with a reagent containing detergent to lyse red blood cells, and the mixture was applied to an immunochromatography strip. The lysate was observed to migrate in the strip and was washed away by the substrate buffer. When the sample contained the specific IgE, the antigen-specific IgE line was clearly observed on the strip macroscopically.ResultsResults were obtained 20 minutes after the application of hemolysed blood sample to immunochromatography, and these results showed positive correlation with those obtained by the AlaSTAT system, which is one of the popular assay kits for specific IgE. The results were not affected significantly by the hematocrit value of the blood sample, by the kind of anticoagulant in the blood collection tube, or by the concentration of the total IgE, provided it was lower than 20000IU/ml.ConclusionsThese results indicate that our system is applicable for point-of-care testing for antigen-specific IgE

Changes in bacterial number at different sites of oral cavity during perioperative oral care management in gastrointestinal cancer patients: preliminary study

Objective: The objective of this study was to clarify differences in bacterial accumulation between gastrointestinal cancer patients who underwent severely invasive surgery and those who underwent minimally invasive surgery. Material and Methods: We performed a preliminary investigation of gastrointestinal cancer patients who were treated at the Department of Surgery, Takarazuka Municipal Hospital, from 2015 to 2017 (n=71; 42 laparoscopic surgery, 29 open surgery) to determine changes in bacterial numbers at different sites of the oral cavity (tongue dorsum, gingiva of upper anterior teeth, palatoglossal arch), as well as mouth dryness and tongue coating indices. Specifically, patients received professional tooth cleaning (PTC), scaling, tongue cleaning, and self-care instruction regarding tooth brushing from a dental hygienist a day before the operation. Professional oral health care was also performed by a dental hygienist two and seven days after surgery. Oral bacteria numbers were determined using a bacterial counter with a dielectrophoretic impedance measurement method. Results: The number of bacteria at all three examined sites were significantly higher in the open surgery group when compared to the laparoscopic surgery group on the second postoperative day. Relevantly, bacterial count in samples from the gingiva of the upper anterior teeth remained greater seven days after the operation in patients who underwent open surgery. Furthermore, the dry mouth index level was higher in the open surgery group when compared to the laparoscopic surgery group on postoperative days 2 and 7. Conclusions: Even with regular oral health care, bacterial numbers remained high in the upper incisor tooth gingiva in gastrointestinal cancer patients who received open surgery. Additional procedures are likely needed to effectively reduce the number of bacteria in the gingival area associated with the upper anterior teet

回転振盪下での動的細胞接着能測定法の開発

The purpose of this study was to develop a method for determining the adhesive ability of cultured cells. The method differs from the conventional methods, and considers the blood stream on the cell adhesion. Also, to keep the cell suspension to be uniform, the procedure was conducted by rotary shaking (120 rpm) the 1X10⁶ cultured cells/10 mL in a 10 cm diameter laboratory dish at 37℃ using a bio-shaker. Measurements of the number of cells remaining in the supernatant were performed over time by using the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5- diphenyl-2H-tetrazolium bromide) method. The adhesive ability of the cells was determined as the time when 50% of the cells had been floating in the supernatant. Four cell lines (8505C, human thyroid carcinoma ; 8305C, human thyroid carcinoma ; A431, human epidermoid carcinoma ; CHO-K1, Chinese hamster ovary) were distinguishable for the adhesive ability (113.1, 131.6, 33.2 and 221.1 min, respectively). For example, by trypsin treatment to harvest nearly all the cell from the laboratory dish bottoms, general reaction times (8505C, 3 min ; 8304/C, 1.5 min ; A431, 7 min ; CHO-K1, 1.5 min) are reverse order above. These results do not contradict the conventional methods. This method is simple and does not need skilled techniques unlike the conventional methods

Possible utilization of salivary IFN-γ/IL-4 ratio as a marker of chronic stress in healthy individuals

Introduction: Several studies show that psychological stress reduces Th1/Th2 ratio in blood samples. However, evidence is scarce regarding the cytokine alterations during stress in saliva. We investigated the influence of chronic stress on Th1/Th2 ratio and cytokine profiles in the saliva of healthy individuals. Further, we examined the associations of the salivary cytokine levels with sleep and attention problems, which are closely related with psychological stress. / Methods: Salivary levels of 27 cytokines were measured by multiplex bead array assays in 31 healthy young individuals (health science students and hospital staff consisting of 11 men and 20 women, mean age [standard deviation] =21.5 [0.8] years). The Kessler Psychological Distress scale (K10) and Athens Insomnia Scale (AIS) were administered to assess subjective chronic psychological stress and sleep problems. Further, participants were asked to wear Actigraph GT3X accelerometers for 3 days to assess the total sleep time. Attention problems were assessed by administering the Integrated Visual and Auditory Continuous Performance Test (IVA-CPT). / Results: Thirteen cytokines with >80% detectable results were included in the main analyses. The IFN-gamma/IL-4 ratio, which is a commonly used index for Th1/Th2 ratio, showed significant negative correlations with the K10 and AIS scores. None of the cytokines were significantly associated with sex, body mass index, sleep index measured by Actigraph, or IVA-CPT scores. / Conclusion: Chronic stress may be associated with alterations of the Th1/Th2 balance in salivary cytokine production. IFN-gamma/IL-4 ratio in saliva may serve as a potential biomarker of chronic stress in healthy individuals.ArticleNeuropsychopharmacology reports 41(1) : 65-72(2021)journal articl

Liver fluke infections by Amphimerus sp. (Digenea: Opisthorchiidae) in definitive and fish intermediate hosts in Manabí province, Ecuador

This work is licensed under a Creative Commons Attribution 4.0 International License.Amphimerus sp. is a fluke that dwells in the biliary tracts of vertebrate definitive hosts including humans, domestic, and wild mammals in Latin America. Opisthorchiid liver infections are rarely studied in the Americas confirming its status as a neglected tropical disease. In Ecuador, small trematode eggs were reported in human cases from the province of Manabí in 1949, and recently, Amphimerus sp. adults were recovered from human and reservoir hosts in the province of Esmeraldas. Due to the lack of research on the infectious sources of Amphimerus sp. in the continent, we have developed a series of epidemiological studies with parasitological and molecular techniques to elucidate the endemicity of opisthorchiid fluke infections. We developed a cross-sectional study in three communities at Pedro Pablo Gómez parish in the province of Manabí, Ecuador. We examined a total of 176 fecal samples to detect opisthorchiid eggs, and four fish species to find opisthorchiid metacercariae. To study adult worms, we treated and purged seven patients in a family and dissected the livers of a dog and a cat infected. We observed morphological features of adults and metacercariae and used polymerase chain reaction with restricted fragment length polymorphism (PCR-RFLP) and DNA sequencing of a section of the ITS2 gene for identification. Small trematode eggs were detected in 63 (35.8%) out of 176 fecal samples of residents in the three study sites. Adult opisthorchiid flukes were recovered from human patients, a dog and a cat, and they were morphologically and molecularly identified as Amphimerus sp. Opisthorchiid metacercariae were also identified molecularly as Amphimerus sp. in four fish species, i.e., Rhoadsia altipinna, Bryconamericus bucay, Andinoacara rivulatus, and Piabucina aureoguttata. Metacercariae of the heterophyid Haplorchis pumilio were also found in the four fish species examined. This is the first study to confirm the current endemicity of Amphimerus sp. in Pedro Pablo Gómez, Manabí, Ecuador. The adult worms isolated here shared morphological characteristics with previous Amphimerus sp. descriptions and were molecularly similar to Amphimerus sp. described in the province of Esmeraldas. Moreover, this study is the first to document four fish species as infection sources of Amphimerus sp. detected via a molecular protocol targeting the metacercariae of the parasite. Fish species identified here should be targeted for public health campaigns to avoid further human liver-fluke infections by Amphimerus sp. or potential intestinal-fluke infections by H. pumilio or others.Japan Society for the Promotion of Science (25305011 and 16H05820)Emerging and Re-emerging Infectious Diseases from the Ministry of Health, Labour and Welfare, Japanese government (H23-Shinko-ippan-014 and H26-Shinko-ippan-009)Japan Agency for Medical Research and Development (20fk0108136j0101)Dirección General de Investigaciones, Universidad de las Américas, Quit

フォトダイオードアレイ検出HPLC を用いた微分クロマトグラフ法によるカフェインの定量

We assessed caffeine levels in bathwater and diluted urine samples by derivative processing of chromatograms obtained through the use of HPLC with photodiode array detection. Chromatograms were differentiated on the basis of d（Abs）/d（min）．The peak position of the target analyte, caffeine, was observed to shift depending on the derivative order applied, but was constant when applying the same derivative order, such that retention times were constant even after differentiation. When the method detection limit was examined using standard solutions, the varying absorption of acetonitrile during a linear gradient elution from 1 to 100 % acetonitrile was shown to have an effect, such that the linearity of the peak height deteriorated if caffeine was present at low concentrations. Recovery of caffeine from real samples was optimized in the case of both linear gradient elution and isocratic elution（ 10 % acetonitrile）when applying the second derivative and using a detection wavelength of 259 nm. Interestingly, this does not correspond to the maximum absorption of caffeine. Thus the optimum wavelength for use with this differential chromatographic method is one that is not affected by the absorbance of the eluent and at which the target analyte shows significant absorbance, even if it is not the maximum absorbance. Such conditions were found to be necessary for the high-sensitivity measurement of caffeine. It is believed that this method is applicable to the HPLC analysis of compounds other than caffeine and this technique is expected to be effective when applied to an automated analyzer and should be helpful to analysts unfamiliar with HPLC.フォトダイオードアレイ検出HPLC を用いた微分クロマトグラフ法による希釈尿あるいは風呂水からのカフェインの回収について検討した。通常のクロマトグラムを微分（d（Abs）/d（min））し，微分クロマトグラムを得る。目的の化合物のピーク位置は，微分次数によるずれるが，同じ微分次数で比較すると一定しており，保持時間による定性能は微分後も維持されていた。標準溶液を用いた定量下限値は，リニアグラジエント溶出（1 → 100 ％アセトニトリル）では溶離液として用いたアセトニトリルの吸収が影響を与え，カフェインが低濃度の場合にはピーク高さとの直線性が悪化した。実試料からのカフェインの回収は，波長259 nm，2 次微分実行時にリニアグラジエント溶出，アイソクラティック溶出（10 ％アセトニトリル）共最適であった。この条件は必ずしもカフェインが極大吸収を持つ波長，微分次数ではない。本結果は微分クロマトグラフ法での検出条件として，1）溶離液吸収の影響を受け難い波長を選択すること，2）目的物質であるカフェインの吸光度が極大でなくてもある程度大きいことが高感度測定に必要であることを示す。今回の結果はカフェインのみならず，他の化合物をHPLC 分析する場合，特にHPLC に不慣れな使用者が条件検討する場合や自動分析装置に応用する場合にも利用可能な知見であると考えられる

Multifrequency VLBI Observations of the Broad Absorption Line Quasar J1020+4320: Recently Restarted Jet Activity?

This paper reports very-long-baseline interferometry observations of the radio-loud broad absorption line (BAL) quasar J1020+4320 at 1.7, 2.3, 6.7, and 8.4 GHz using the Japanese VLBI network (JVN) and European VLBI network (EVN). The radio morphology is compact with a size of ~10 pc. The convex radio spectrum is stable over the last decade; an observed peak frequency of 3.2 GHz is equivalent to 9.5 GHz in the rest frame, suggesting an age of the order of ~100 years as a radio source, according to an observed correlation between linear size and peak frequency of compact steep spectrum (CSS) and giga-hertz peaked spectrum (GPS) radio sources. A low-frequency radio excess suggests relic of past jet activity. J1020+4320 may be one of the quasars with recurrent and short-lived jet activity during a BAL-outflowing phase.Comment: 7 pages, 2 figures, 2 tables, accepted for publication in PAS

Negative Correlation between Serum Cytokine Levels and Cognitive Abilities in Children with Autism Spectrum Disorder

Evidence suggests that cytokines may be one of the major factors influencing cognitive development in those with autism spectrum disorder (ASD). To shed light on the neural and cognitive mechanisms of ASD, we investigated the association between peripheral cytokine levels and cognitive profiles in children with ASD. The serum levels of 10 cytokines (granulocyte macrophage colony-stimulating factor, interferon (IFN)-γ, interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, and tumor necrosis factor-α) were examined in 14 children with ASD using the Human Ultrasensitive Cytokine Magnetic 10-Plex Panel for the Luminex platform. The Wechsler Intelligence Scale for Children (WISC) was administered to each subject, and the relationships between WISC scores and serum levels of the cytokines were examined. The full-scale intelligence quotient (IQ) was significantly negatively correlated with the levels of IL-6 (Spearman’s rank, p < 0.0001, false discovery rate q < 0.01). The levels of IL-6 and IFN-γ showed significant negative correlations with the verbal comprehension index (p < 0.001, q < 0.01) and working memory index (p < 0.01, q < 0.05), respectively. No other cytokines were significantly correlated with full-scale IQ or with any of the subscale scores of the WISC. The present results suggest negative correlations of IL-6 and IFN-γ levels with cognitive development of children with ASD. Our preliminary findings add to the evidence that cytokines may play a role in the neural development in ASD.ArticleJOURNAL OF INTELLIGENCE. 5(2):19 (2017)journal articl