Microbiome analysis of contact lens care solutions and tear fluids of contact lens wearers: Possible involvement of streptococcal antigens in allergic symptoms related to contact lens wear.

The present study elucidated the pathogenesis of allergic symptoms (AS) related to contact lens (CL) wear by assaying CL care solutions in lens storage cases and tears from subjects with AS using molecular biology techniques. A total of 15 CL storage cases were collected from subjects with AS (n=9) and healthy, asymptomatic control CL wearers (n=6). Bacterial populations in CL care solutions and tears were assayed by culture and 16S rDNA sequencing. Histamine levels in tears were measured by high­performance liquid chromatography. Western blot analysis was performed to identify the bacteria recognized by tear IgE from subjects with AS. No significant differences were found in the culture results between the subjects with AS and asymptomatic subjects. Histamine was detected in 2 subjects with AS. Meta­16S rDNA sequencing identified a cluster of 4 subjects with AS that were distinguished from others by principal coordinate analysis. Detailed population analysis revealed that the abundance of Gram­positive bacteria in the microbiomes of CL care solutions used by the subjects with AS were higher than those of asymptomatic subjects (42.24±9.47 vs. 16.85±22.76% abundance). Among these, Streptococcus was the dominant genus (12.1­18.3% abundance). Tear microbiome analysis revealed that the abundance of Streptococcus in the subjects with AS was significantly higher than that in other subjects (19.02±5.50 vs. 3.08±3.35%, P<0.01). Western blot analysis demonstrated that the tear IgE in all subjects with AS reacted with Streptococcus (100%), but not with Staphylococcus. On the whole, these results provide novel insight into the pathogenesis of AS and identify Streptococcus as an important factor in AS associated with CL wear.

Effect of thymoquinone on Fusobacterium nucleatum­associated biofilm and inflammation.

Periodontitis affects oral tissues and induces systemic inflammation, which increases the risk of cardiovascular disease and metabolic syndrome. Subgingival plaque accumulation is a trigger of periodontitis. Fusobacterium nucleatum (FN) contributes to subgingival biofilm complexity by intercalating with early and late bacterial colonizers on tooth surfaces. In addition, inflammatory responses to FN are associated with the progression of periodontitis. Nigella sativa Lin. seed, which is known as black cumin (BC), has been used as a herbal medicine to treat ailments such as asthma and infectious diseases. The current study examined the inhibitory effect of BC oil and its active constituents, thymol (TM) and thymoquinone (TQ), on FN­associated biofilm and inflammation. FN­containing biofilms were prepared by co­cultivation with an early dental colonizer, Actinomyces naeslundii (AN). The stability and biomass of FN/AN dual species biofilms were significantly higher compared with FN alone. This effect was retained even with prefixed cells, indicating that FN/AN co­aggregation is mediated by physicochemical interactions with cell surface molecules. FN/AN biofilm formation was significantly inhibited by 0.1% TM or TQ. Confocal laser scanning microscopy indicated that treatment of preformed FN/AN biofilm with 0.01% of BC, TM or TQ significantly reduced biofilm thickness, and TQ demonstrated a cleansing effect equivalent to that of isopropyl methylphenol. TQ dose­dependently suppressed TNF­α production from a human monocytic cell line, THP­1 exposed to FN, yet showed no toxicity to THP­1 cells. These results indicated that oral hygiene care using TQ could reduce FN­associated biofilm and inflammation in gingival tissue.

Ethanolamine utilization supports Clostridium perfringens growth in infected tissues.

Clostridium perfringens possesses the ethanolamine (EA) utilization (eut) system encoded within the eut operon, which utilizes the EA as a carbon, nitrogen and energy source. To determine the role of the eut system in C. perfringens growth, an in-frame deletion of the eutABC genes was made in strain HN13 to generate the eutABC-deleted mutant strain HY1701. Comparison of HN13 and HY1701 growth in media supplemented with 1.0% glucose and/or 1.0% EA showed that glucose enhanced the growth of both strains, whereas EA enhanced HN13 growth, but not that of HY1701, indicating that the eut system is necessary for C. perfringens to utilize EA. The two-component regulatory system EutVW is needed to induce eut gene expression in response to EA whereas the global virulence regulator VirRS differentially controlled eut gene expression depending on glucose and EA availability. To assess the role of the eut system in vivo, an equal number of HN13 and HY1701 cells were injected into the right thigh muscles of mice. Mice infected with HY1701 showed fewer symptoms than those injected with HN13. The mortality rate of mice infected with HY1701 tended to be lower than for mice infected with HN13. In addition, in infected tissues from mice injected with a mixture of HN13 and HY1701, HN13 outnumbered HY1701. PCR screening demonstrated that C. perfringens isolated from gas gangrene and sporadic diarrhea cases carried both eut genes and the perfringolysin O gene (pfoA) as well as the phospholipase C gene (plc). However, pfoA was not detected in isolates from food poisoning patients and healthy volunteers. Culture supernatants prepared from HN13 grown in media containing 7.5% sheep red blood cells induced significantly higher eutB expression levels compared to those from plc- and/or pfoA-deletion mutants. Together, these results indicate that the eut system plays a nutritional role for C. perfringens during histolytic infection.

[Experimental study of detectability of simulated lung nodule on computed radiography system using various X-ray beam qualities].

The purpose of this study was to discuss the optimal X-ray beam quality for detection of simulated lung nodule on a computed radiography system. We set up four types of X-ray beam quality (90 kV, 120 kV, 150 kV, and 120 kV + gadolinium filter), and kept the incident dose on the patient at 0.3 mGy. A receiver operating characteristic (ROC) analysis and a granularity measurement were used to evaluate the relationship between the detection of a low-contrast object and X-ray beam quality. As a result, the areas under the ROC curve (AUC) of 90 kV and 120 kV + gadolinium filter were significantly superior to those of 120 kV and 150 kV (p<0.05). However, a significant difference was not observed between 90 kV and 120 kV + gadolinium filter, 120 kV and 150 kV. The order of the granularity values gave good agreement with the results of visual evaluation. In conclusion, we considered that the optimal X-ray beam quality was 90 kV or 120 kV + gadolinium filter.

[A Study of uncertainty factors in cross calibration of dosimeter for diagnostic X-rays].

Although patient exposure has been increasing in recent years, few institutions have dosimeters and are able to ascertain patients' exposure dose. Internationally, however, it is necessary to adopt safety levels for patient exposure doses, and guidance levels have been introduced. Therefore, the need for measurement in areas of x-ray diagnosis has been increasing. As a result, several societies concerned with radiation dose have been endeavoring to establish a calibration system of radiation measurement and a dosimeter calibration system, which are the basics of radiation protection. Ten regional centers for standardization of doses in x-ray diagnosis were established and have begun trials relating to dosimeter cross calibration. Our institution, as one of these centers, has instituted a trial. In this study, the cross-calibration field, the reliability of the cross-calibration skill of our regional center, and the standard uncertainty of cross calibration were investigated. As a consequence of the investigation, it was determined that our cross-calibration field follows the protocol of the Japanese Society of Radiological Technology, the difference between calibration factor/cross calibration factor obtained by JQA and our regional center is within 2.5%, and the expanded uncertainty of our cross calibration is about 7.2% (k=2).

[Measurement of half-value layer in mammography].

The half-value layer (HVL) of an X-ray beam for film-screen mammography is considered an important parameter for image quality and patient dose. Thus, HVL must be measured in accordance with The Manual of Accuracy for Mammography printed by the Japanese Society of Radiological Technology. The manual prescribes exactly the geometry of measurement, chamber position of measurement in the field, selection of chamber, and so on. However, the measurement of HVL is difficult in the actual clinical setting. This study examined the results of failure to perform the measurement of HVL in accordance with the manual for measuring HVL in the clinical setting. The investigation indicated that serious problems do not arise when measuring HVL for routine quality control even if the chamber in the field is not always set according to the manual and if a chamber for radiotherapy or diagnosis is used that is not recommended for soft X-ray by the manual.

[Patient skin dose in interventional radiology using radiochromic dosimetry film].

Various types of X-ray examinations are currently being carried out for the purpose of diagnosis. However, since dose limits for contamination by medical examinations have not been set, management of dose measurements and contamination records is called for. With increasing use of the IVR technique, reports of radiation injury and the symptoms associated with it have become more common. To advance our understanding of this situation and to reduce contamination, it is necessary to carry out contamination management. The reflection film on which colors are formed by irradiating X-rays has recently come into use. Dose measurement is possible with the use of this film, and, because effective results can be obtained as a result of performing fundamental examinations, the film actually provides dose measurements for the IVR technique. Another benefit is that maximum patient skin dose and dose distribution can be determined in addition to dose measurement. Moreover, since various methods were examined in this study, the method of dose evaluation is also reported for those wishing to employ it in the clinical setting.

Alternative splicing variants of the human DBL (MCF-2) proto-oncogene.

The DBL (MCF-2) proto-oncogene is a prototype guanine nucleotide exchange factor (GEF) that modulates the Rho family of GTPases. In this communication we describe the isolation of three novel splicing variants of Dbl. The prototype Dbl gene (designated var.1 here) contains 25 exons, while splicing variant 2 (var.2) lacks exons 23 and 24. Var.3 contains additional 3 exons from 5(')-UTR in place of exon 1, while var.4, var.2, and var.3 contain a 48bp insertion between exons 10 and 11, resulting in the insertion of 16 amino acids. We found that var.1 was expressed only in brain, whereas var.3 was expressed in heart, kidney, spleen, liver, and testis, and var.4 in brain, heart, kidney, testis, placenta, stomach, and peripheral blood. The Dbl protein was detectable in brain, heart, kidney, intestine, muscle, lung, and testis. An assay for GEF activity revealed that the var.2 exhibits decreased GEF activity towards Cdc42, var.3 exhibits a weak but significant activity toward Rac1 and Cdc42, var.4 exhibits significant activity toward RhoA and Cdc42, while var.1 exhibits no activity toward RhoA, Rac1, or Cdc42. In summary, we describe 4 splicing variants of the human DBL proto-oncogene that show different tissue distributions and GEF specificities.

[Radiation exposure in CT].

The use of CT has increased rapidly in the past two decades, in part because of the development of helical CT and multidetector row CT. The benefits obtained by CT are not, however, without risks. This paper emphasizes the importance of keeping radiation doses during CT procedures as low as reasonably possible, especially for pediatric and small adult patients. In order to evaluate the risk of exposure, precise dosimetry is important. This paper introduces a new film dosimetry system that can be used easily in any institution and with any CT apparatus.

[Medical exposure from computed tomographic examination -a recommendation of dosimetry by film method-].

Already, the 30 year have passed since that the CT scanner was developed by Hunsfield. And now above 16,000 sets are at work. On the other hand, as the improvements of image reconstruction algorithm and variations os scan method, the medical exposure is increasing. For example, in the case of computed radiography, medical exposure is stability by using an auto exposure controller but in the case of CT, there is no unit and usually the exposure condition of CT examination are used that of recommended an exposure condition by maker. So the medical exposure greatly differs according to the operators or the kinds of CT scanner. In addition, there is the close connection between the x-ray intensity and the image quality. Especially, in the case of CT scanner there is no saturation of x-ray intensity like the optical density of film screen system because the digital radiographic systems have the wide dynamic range. So the increase in x-ray intensity makes a contribution to improvement of the image quality. But the increase in x-ray intensity makes a contribution to the increase in medical exposure. Therefore it is very important that the CT operators grasp the medical exposure per each CT examination and patient. There are many methods of medical exposure dosimetry on CT examination. But almost all the methods except the film method are impossible to get the medical exposure per each CT examination and patient. So in this paper the film method is recommended. Because the film method is low-priced, simple, easy and immediately shows the medical exposure per each CT examination and patient. Furthermore, it is confirmed that the film method is getting the correct medical exposure because that values gotten by the film method and ionization chamber are almost equivalent.