Characterization of epitopes of human monoclonal antibodies against cytomegalovirus glycoprotein B for neutralization and antibody-dependent phagocytosis.

As congenital cytomegalovirus (CMV) infections are the leading non-genetic cause of sensorineural hearing loss and significant neurological disabilities in children, the development of CMV vaccines should be given the highest public health priority. Although MF59-adjuvanted glycoprotein B (gB) vaccine (gB/MF59) is safe and immunogenic, its efficacy in terms of protection from natural infection was around 50 % in clinical trials. Although gB/MF59 induced high antibody titers, anti-gB antibodies contributed little to the neutralization of infection. Recent studies have found that non-neutralizing functions, including antibody-dependent phagocytosis of virions and virus-infected cells, are likely to play important roles in pathogenesis and vaccine design. Previously, we isolated human monoclonal antibodies (MAbs) that reacted with the trimeric form of gB ectodomain and found that preferential epitopes for neutralization were present on Domains (Doms) I and II of gB, while there were abundant non-neutralizing antibodies targeting Dom IV. In this study, we analyzed the phagocytosis activities of these MAbs and found the following: 1) MAbs effective for phagocytosis of the virions targeted Doms I and II, 2) the MAbs effective for phagocytosis of the virions and those of virus-infected cells were generally distinct, and 3) the antibody-dependent phagocytosis showed little correlation with neutralizing activities. Taking account of the frequency and levels of neutralization and phagocytosis, incorporation of the epitopes on Doms I and II into developing vaccines is considered desirable for the prevention of viremia.

The Body Fat Percentage Rather Than the BMI Is Associated with the CD4 Count among HIV Positive Japanese Individuals.

Maintenance of the cluster of differentiation 4 (CD4) positive lymphocyte count (CD4 count) is important for human immunodeficiency virus (HIV) positive individuals. Although a higher body mass index (BMI) is shown to be associated with a higher CD4 count, BMI itself does not reflect body composition. Therefore, we examined the association of body weight, body composition and the CD4 count, and determined the optimal ranges of CD4 count associated factors in Japanese HIV positive individuals. This cross-sectional study included 338 male patients treated with antiretroviral therapy for ≥12 months. Multiple logistic regression analysis was used to identify factors significantly associated with a CD4 count of ≥500 cells (mm3)-1. The cutoff values of factors for a CD4 ≥ 500 cells (mm3)-1 and cardiovascular disease risk were obtained by receiver operating characteristic curves. Age, body fat percentage (BF%), nadir CD4 count, duration of antiretroviral therapy (ART), years since the HIV-positive diagnosis and cholesterol intake showed significant associations with the CD4 count. The cutoff value of BF% for a CD4 ≥ 500 cells (mm3)-1 and lower cardiovascular disease risk were ≥25.1% and ≤25.5%, respectively. The BF%, but not the BMI, was associated with CD4 count. For the management of HIV positive individuals, 25% appears to be the optimal BF% when considering the balance between CD4 count management and cardiovascular disease risk.

Risk Factors for Falls in Patients with Alzheimer Disease: A Retrospective Study of Balance, Cognition, and Visuospatial Ability.

AIM: This study aimed to determine the main risk factors for falls in patients with Alzheimer disease (AD) by comparing balance, cognition, and visuospatial ability between those who have experienced a fall and those who have not. METHODS: Forty-seven AD patients were admitted to a ward for patients with dementia (22 men and 25 women). The balance of patients was evaluated using the Functional Reach Test (FRT), the one-leg standing duration, and the Timed Up and Go (TUG) test. The Mini-Mental State Examination-Japanese (MMSE-J) was used to evaluate cognition. For visuospatial ability assessment, the Clock-Drawing Test (CDT) as well as overlapping figure identification and shape discrimination in the Visual Perception Test for Agnosia (VPTA) were used. The patients were allocated to either the fall group or the nonfall group based on their history of falls in the past year. The relationships between patients' characteristics and evaluation outcomes were compared and examined. Logistic regression analysis was performed using a fall as the objective variable. The area under the curve (AUC) and the cutoff value were calculated. RESULTS: Of the 47 participants, 22 had experienced falls within the past year (46.8%). The results of the FRT, one-leg standing duration, the TUG, the CDT, and the VPTA were significantly lower in the fall group. No significant difference between the MMSE-J scores of the fall group and those of the nonfall group was observed. The results of the logistic regression analysis indicated that falls in AD patients were significantly associated with the FRT. It was found that a shorter FRT distance (cm) had a significant impact on falls. For the FRT, the fall-related AUC was 0.755. At a cutoff value of 24.5 cm, the level of sensitivity was 68.0%, and the level of specificity was 77.3%. CONCLUSIONS: The findings of this study indicate that balance and visuospatial abilities are risks factors for falls in AD patients. In contrast, cognitive impairment was not a risk factor for falls. It was demonstrated that the FRT could be an appropriate risk predictor for falls in AD patients. In particular, falls in AD patients were strongly affected by a reduced dynamic balance.

Quantitative and Qualitative Analyses of the Clock Drawing Test in Fall and Non-Fall Patients with Alzheimer's Disease.

AIM: The clock drawing test (CDT) is widely used as a visual spatial ability test and screening test for dementia patients. The appearance frequency of qualitative errors obtained through the qualitative analysis of CDT may be related to the participant's falls. The aim of this study was to clarify the difference in the number of people who presented with qualitative errors in the CDT between a fall and non-fall group of patients with Alzheimer's disease (AD). METHODS: The CDT was implemented for 47 patients with AD. A quantitative analysis was conducted, and a qualitative analysis was performed for errors. The patients were divided into two groups based on their history of falls over the past year. The results of the CDT quantitative analysis were tested using the Mann-Whitney U test, and Fisher's exact test was employed to determine the difference in the number of people who presented with error types between the two groups (fall group, non-fall group) in the CDT qualitative analysis. RESULTS: In the quantitative analysis, a significant difference was found for the total scores, with the total CDT score of the fall group (n = 22) significantly lower than that of the non-fall group (n = 25) (p = 0.006, effect size: φ = 0.40). In the qualitative analysis, a significantly higher number of patients in the fall group than in the non-fall group presented with a conceptual deficit (p =0.001, φ = 0.51). No differences were found in the number of patients in the two groups who presented with the other five error types. CONCLUSIONS: These results showed that a lower score in the CDT quantitative analysis might suggest an increased risk of falls. It was also clarified that a larger number of patients in the fall group than in the non-fall group presented with a conceptual deficit of the qualitative error types in the CDT. Therefore, these results suggest that the appearance of a conceptual deficit may be an index for the selection of patients with AD prone to falling when implementing fall prevention measures.

Resistance to Thyroid Hormone Complicated with Type 2 Diabetes and Cardiomyopathy in a Patient with a TRß Mutation.

Resistance to thyroid hormone (RTH) is a genetic disorder characterized by reduced tissue responsiveness to thyroid hormone. We herein describe a 60-year old man who presented with the clinical features of cardiomyopathy, diabetes mellitus and elevated thyroid hormones with unsuppressed thyroid stimulating hormone. A genetic analysis of thyroid hormone receptor (TR) revealed a missense mutation (A268D) in the TRß gene. Clinical manifestations of RTH may be variable due to different tissue distributions of TR subtypes and different actions of mutant receptors. The current case demonstrates that patients with a TRß mutation may have impaired his glucose metabolism and a reduced cardiac function, although patients appear clinically euthyroid.

Ig L-chain shuffling for affinity maturation of phage library-derived human anti-human MCP-1 antibody blocking its chemotactic activity.

Monocyte chemotactic protein-1 (MCP-1, CC-chemokine ligand 2; CCL2) is involved in the development of various forms of chronic inflammations. Employing the naive human single-chain Fv displaying phage library, we established seven MCP-1-specific scFvs. The MC8 and MC32 clones exhibited blocking activity for the MCP-1-induced chemotaxis of THP-1 cells, in spite of their monovalency. The analysis of V gene usage showed that all clones bore the identical Vh1 gene, IGHV1-24*01, with variable DJ joining sequences, while their Vl usage was relatively varied, suggesting the preferential contribution of the Vh gene. Based on these findings, to minimize the deteriorative influences on the MCP-1 specificity of MC32, we aimed to achieve the affinity maturation of MC32 using MC32 L-chain shuffling library and select MC32 variants. Most MC32 variants increased their affinity by reducing the k(off) value with no influence of the antigen specificity. MC32 variants #22 or #56 showed approximately 15-fold higher affinity than MC32, indicating that the L-chain shuffling library is useful if the Vh is dominantly involved in the determination of the antigen specificity.

Metabolism of 5'alpha,8'-cycloabscisic acid, a highly potent and long-lasting abscisic acid analogue, in radish seedlings.

We synthesized 5'alpha,8'-cycloabscisic acid (CycloABA), a highly potent and long-lasting abscisic acid (ABA) analogue, by a different method from that reported before. CycloABA fed to radish seedlings had more metabolic tolerance than ABA. The major metabolite of CycloABA was the glucose conjugate, which was the minor metabolite of ABA. The 8'-hydroxylated metabolite and its cyclized isomer, which were major metabolites of ABA, were not found as metabolites of CycloABA. The present results suggest that the highly potent and long-lasting activity of CycloABA is caused by resistance to ABA 8'-hydroxylase, and that CycloABA is partially metabolized to the glucose conjugate by ABA glucosyltransferase.

Current hair mercury levels in Japanese: survey in five districts.

To understand the current Japanese hair mercury levels, we planned a survey of hair mercury among the general populations of different regions in Japan. The present paper, as the first report of the survey, summarized the results obtained in five districts, Minamata, Kumamoto, Tottori, Wakayama and Chiba. Hair samples were collected at beauty salons, barbershops and primary schools in each district with questionnaires on age, sex, amount and species of fish usually consumed, hair-dyed and artificial hair waving "permanent wave." The total mercury levels of 3686 hair samples collected were analyzed by an oxygen combustion-gold amalgamation method. The geometric mean of the total mercury concentration was significantly higher in males than in females, i.e., 2.55 microg/g and 1.43 microg/g, respectively. The sex difference was also observed on hair samples without artificial waving, i.e., 2.64 microg/g and 1.64 microg/g, respectively. The geometric mean in each district varied from 2.23 to 4.79 microg/g for males and from 1.23 to 2.50 microg/g for females. The average hair mercury levels were highest in Chiba among the five districts both in males and females. A multiple regression analysis revealed a significant correlation of the mercury level with age, sex, amount of daily fish consumption, tuna and bonito as usually consumed fish, artificial waving and Chiba as a residential area. In the laboratory experiment, we found that the treatment of hair samples with a lotion for artificial waving caused a 30%-reduction in the mercury content. Furthermore, longitudinal hair analysis showed a marked difference in the concentration between the hair root and the tip of the hair taken from artificially waved females; higher values were observed at the hair root. These results suggested that artificial waving significantly removes hair mercury and that hair analysis at the hair root should be necessary to estimate an accurate methylmercury exposure for waved persons.