Determination of Nonspherical Morphology of Doxorubicin-Loaded Liposomes by Atomic Force Microscopy

The 3-D morphology of doxorubicin (DOX)–loaded liposomes with a size of circa 100 nm was characterized by atomic force microscopy in an aqueous environment. Prolate liposomes appear in accordance with linear expansion of DOX fiber bundles precipitated inside liposomes. Oblate and concave liposomes were simultaneously observed with increased DOX concentrations; however, their morphologies were not readily determined by 2-D cryo-TEM imaging. Precise data analysis of the 3-D parameters of each liposome allowed semiquantitative evaluation of the transformation of spherical liposomes into nonspherical—prolate, oblate, and concave liposomes. In addition, nonspherical liposomes became spherical on the replacement of the liposomal outer phase consisting of a sucrose solution, with water and subsequent water influx. All spherical liposomes transformed into oblate and concave liposomes with a return to hyperosmotic conditions, when transferred from water to sucrose solution. Furthermore, the concave liposomes did not appear under DOX incubation conditions (65°C), which could be due to the amorphous and supersaturated DOX inside the liposomes that restrained liposomal shrinkage. As atomic force microscopy has improved our ability to image 3-D morphologies of liposomes in various conditions, it is an alternative analytical tool to cryo-TEM and may have future applications in regulatory tests for quality control and assurance.     

Lifestyle factors and social ties associated with the frequency of laughter after the Great East Japan Earthquake: Fukushima Health Management Survey

Purpose

Although mental health problems such as depression after disasters have been reported, positive psychological factors after disasters have not been examined. Recently, the importance of positive affect to our health has been recognised. We therefore investigated the frequency of laughter and its related factors among residents of evacuation zones after the Great East Japan Earthquake of 2011.

Methods

In a cross-sectional study on 52,320 participants aged 20 years and older who were included in the Fukushima Health Management Survey in Japan’s fiscal year 2012, associations of the frequency of laughter with changes in lifestyle after the disaster, such as a changed work situation, the number of family members, and the number of address changes, and other sociodemographic, psychological, and lifestyle factors were examined using logistic regression analysis. The frequency of laughter was assessed using a single-item question: “How often do you laugh out loud?”

Results

The proportion of those who laugh almost every day was 27.1%. Multivariable models adjusted for sociodemographic, psychological, and lifestyle factors demonstrated that an increase in the number of family members and fewer changes of address were significantly associated with a high frequency of laughter. Mental health, regular exercise, and participation in recreational activities were also associated with a high frequency of laughter.

Conclusion

Changes in lifestyle factors after the disaster were associated with the frequency of laughter in the evacuation zone. Future longitudinal studies are needed to examine what factors can increase the frequency of laughter.

     

Development of a novel ELISA for detection of anti-A and anti-B antibodies in recipients of ABO-incompatible living donor liver grafts

The survival rate in ABO-incompatible (ABO-I) liver transplantation was much lower than that in ABO-compatible recipients for the early experiences. It is therefore essential to develop the precise and fast measurement of anti-A and anti-B antibodies (Abs) to prevent humoral rejection in ABO-I liver transplantation. Agglutination titer has been the standard method to measure these Abs, but the interpretation of the results is subject to bias. Here, we have developed an objective and quantitative enzyme-linked immunosorbent assay (ELISA) to measure anti-A and anti-B Abs. This test requires only a small amount (10 microl) of recipient's serum. We applied the newly developed ELISA to monitor living donor liver transplant recipients and investigated the correlation between ELISA and agglutination titer. The Spearman's correlation coefficient for Abs ranged from 0.461 to 0.812. Moreover, in one case of humoral rejection, the increase of Abs was detected by ELISA one day earlier than by the agglutination titer. In conclusion, our ELISA method proved useful to detect an increase of anti-A and anti-B Abs titers at an early stage, thereby contributing to a prompt treatment of humoral rejection due to ABO-I.     

A new evaluation method for upper extremity dexterity of patients with hemiparesis after stroke: the 10-second tests

We developed a new, sensitive evaluation method for upper extremity dexterity in patients with hemiparesis after stroke. This evaluation method consists of three types of test: the Finger Individual Movement Test, Hand Pronation and Supination Test, and Finger Tapping Test. In each test, the number of movements that can be performed during a 10-s period is measured. Data obtained by these 10-s tests were compared with those obtained by the Nine-Hole Peg Test (NHPT), and the usefulness of this new method was evaluated. In patients with hemiparesis after stroke, three types of 10-s test and NHPT were performed for the upper extremities on both the affected and unaffected sides. Simple correlation analysis and multiple regression analysis of data obtained by the three types of test and NHPT were performed using the affected/unaffected ratios. Twenty patients participated (54-85 years; 0.23-43.83 months after stroke) with mild upper extremity hemiparesis. Significant correlations exist between Finger Individual Movement Test and NHPT data (r=-0.584, P=0.0068) and between Hand Pronation and Supination Test and NHPT data (r=-0.707, P=0.0005). The multiple regression analysis model using the three types of 10-s test as explanatory variables was significant (P=0.0025) and explained 52.4% of the NHPT results. A significant association was observed between NHPT and the 10-s tests. The 10-s tests require no special examination instruments and can be readily performed in a short period. This evaluation method consisting of the 10-s tests can be a new clinical parameter of dexterity in patients with hemiparesis after stroke.     

Profiling of bacterial flora in crevices around titanium orthodontic anchor plates

OBJECTIVES: The aims of this study were to characterize the microflora in crevices around titanium orthodontic anchor plates using anaerobic culture and molecular biological techniques for bacterial identification, and to compare the microbial composition between crevices around anchor plates and gingival crevices. MATERIAL AND METHODS: Samples from crevices around titanium anchor plates and healthy gingival crevices of 17 subjects (aged 20-29) were cultured anaerobically, and isolated bacteria were identified by 16S rRNA sequencing. RESULTS: The average logarithm colony-forming units/ml were 6.84, 7.51 and 8.88 in healthy anchor plate crevices, inflamed anchor plate crevices and healthy gingival crevices, respectively, indicating that the bacterial density of anchor plate crevices was lower than that of healthy gingival crevices. Of 184 strains isolated from healthy anchor plate crevices of seven subjects, 108 (59%) were anaerobic bacteria, while 73 (40%) were facultative bacteria. Predominant isolates were Gram-negative rods, such as Campylobacter (12%), Fusobacterium (10%) and Selenomonas (10%), and Gram-positive facultative bacteria, such as Actinomyces (17%) and Streptococcus (8.2%). Of 133 strains isolated from inflamed anchor plate crevices of three subjects, 110 (83%) were anaerobic bacteria, while predominant isolates were Gram-negative rods, such as Prevotella (47%), Fusobacterium (33%) and Campylobacter (16%). On the other hand, of 146 strains isolated from healthy gingival crevices of seven subjects, 98 (67%) were facultative bacteria, while 45 (31%) were anaerobic bacteria. Predominant isolates were Gram-positive facultative bacteria, such as Actinomyces (37%) and Streptococcus (20%). CONCLUSIONS: These results suggest that the environment in crevices around titanium orthodontic anchor plates is anaerobic and supportive of anaerobic growth of bacteria, which may trigger inflammation in the tissue around the plates.     

Induction of immune responses and prevention of alveolar bone loss by intranasal administration of mice with Porphyromonas gingivalis fimbriae and recombinant cholera toxin B subunit

INTRODUCTION: Adult periodontitis is initiated by specific periodontal pathogens represented by Porphyromonas gingivalis; however, an effective measure for preventing the disease has not yet been established. In this study, the effectiveness of a vaccine composed of fimbriae of P. gingivalis and recombinant cholera toxin B subunit (rCTB) was evaluated using BALB/c mice. METHODS: Fimbriae and rCTB were co-administered intranasally to BALB/c mice on days 0, 14, 21, and 28. On day 35, mice were sacrificed to determine immunoglobulin levels in serum, saliva, and nasal and lung extracts by enzyme-linked immunosorbent assay. The prevention effect of the vaccine on P. gingivalis-induced periodontitis in mice was evaluated by measuring alveolar bone loss. RESULTS: The rCTB significantly increased serum immunoglobulin (Ig)A levels when mice were administered with a minimal amount (0.5 microg) of the fimbrial antigen. The adjuvant effect on serum IgG production was indistinct because the minimal amount of the antigen still induced a large amount of IgG. In contrast to systemic responses, a fimbria-specific secretory IgA response was strongly induced by co-administration of rCTB and 0.5 microg fimbriae; the same amount of the antigen alone scarcely induced a response. Histopathological examination revealed IgA-positive plasma cells in the nasal mucosal tissue but no observable mast cells in the area. In addition, nasal administration of the fimbrial vaccine significantly protected the mice from P. gingivalis-mediated alveolar bone loss. CONCLUSION: Nasal vaccination with a combination of fimbriae and rCTB can be an effective means of preventing P. gingivalis-mediated periodontitis.