Infectious virus shedding duration reflects secretory IgA antibody response latency after SARS-CoV-2 infection.

Infectious virus shedding from individuals infected with severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) is used to estimate human-to-human transmission risk. Control of SARS-CoV-2 transmission requires identifying the immune correlates that protect infectious virus shedding. Mucosal immunity prevents infection by SARS-CoV-2, which replicates in the respiratory epithelium and spreads rapidly to other hosts. However, whether mucosal immunity prevents the shedding of the infectious virus in SARS-CoV-2-infected individuals is unknown. We examined the relationship between viral RNA shedding dynamics, duration of infectious virus shedding, and mucosal antibody responses during SARS-CoV-2 infection. Anti-spike secretory IgA antibodies (S-IgA) reduced viral RNA load and infectivity more than anti-spike IgG/IgA antibodies in infected nasopharyngeal samples. Compared with the IgG/IgA response, the anti-spike S-IgA post-infection responses affected the viral RNA shedding dynamics and predicted the duration of infectious virus shedding regardless of the immune history. These findings highlight the importance of anti-spike S-IgA responses in individuals infected with SARS-CoV-2 for preventing infectious virus shedding and SARS-CoV-2 transmission. Developing medical countermeasures to shorten S-IgA response time may help control human-to-human transmission of SARS-CoV-2 infection and prevent future respiratory virus pandemics.

Statin treatment rescues FGFR3 skeletal dysplasia phenotypes.

Gain-of-function mutations in the fibroblast growth factor receptor 3 gene (FGFR3) result in skeletal dysplasias, such as thanatophoric dysplasia and achondroplasia (ACH). The lack of disease models using human cells has hampered the identification of a clinically effective treatment for these diseases. Here we show that statin treatment can rescue patient-specific induced pluripotent stem cell (iPSC) models and a mouse model of FGFR3 skeletal dysplasia. We converted fibroblasts from thanatophoric dysplasia type I (TD1) and ACH patients into iPSCs. The chondrogenic differentiation of TD1 iPSCs and ACH iPSCs resulted in the formation of degraded cartilage. We found that statins could correct the degraded cartilage in both chondrogenically differentiated TD1 and ACH iPSCs. Treatment of ACH model mice with statin led to a significant recovery of bone growth. These results suggest that statins could represent a medical treatment for infants and children with TD1 and ACH.

Changes in acetyl-CoA mediate Sik3-induced maturation of chondrocytes in endochondral bone formation.

The maturation of chondrocytes is strictly regulated for proper endochondral bone formation. Although recent studies have revealed that intracellular metabolic processes regulate the proliferation and differentiation of cells, little is known about how changes in metabolite levels regulate chondrocyte maturation. To identify the metabolites which regulate chondrocyte maturation, we performed a metabolome analysis on chondrocytes of Sik3 knockout mice, in which chondrocyte maturation is delayed. Among the metabolites, acetyl-CoA was decreased in this model. Immunohistochemical analysis of the Sik3 knockout chondrocytes indicated that the expression levels of phospho-pyruvate dehydrogenase (phospho-Pdh), an inactivated form of Pdh, which is an enzyme that converts pyruvate to acetyl-CoA, and of Pdh kinase 4 (Pdk4), which phosphorylates Pdh, were increased. Inhibition of Pdh by treatment with CPI613 delayed chondrocyte maturation in metatarsal primordial cartilage in organ culture. These results collectively suggest that decreasing the acetyl-CoA level is a cause and not result of the delayed chondrocyte maturation. Sik3 appears to increase the acetyl-CoA level by decreasing the expression level of Pdk4. Blocking ATP synthesis in the TCA cycle by treatment with rotenone also delayed chondrocyte maturation in metatarsal primordial cartilage in organ culture, suggesting the possibility that depriving acetyl-CoA as a substrate for the TCA cycle is responsible for the delayed maturation. Our finding of acetyl-CoA as a regulator of chondrocyte maturation could contribute to understanding the regulatory mechanisms controlling endochondral bone formation by metabolites.

Dynamic performance of bracket complexes used in traditional timber structures in Japan.

A bracket complex is a type of bearing system located on the top of a column and transmits the vertical load of the roof to the column. Bracket complexes are often used in Buddhist temples and Shinto shrines constructed using traditional timber structure. The bracket complex, being a combination of timber blocks and beam like elements, is said to have a positive effect on the seismic performance of traditional timber structures. However, this hypothesis has not been theoretically or quantitatively proven. The aim of the present paper is to clarify the dynamic performance of the bracket complex. The investigations of the present study were performed through experiments and analysis. Shaking table tests were conducted on four different types of bracket complexes. The stiffness, natural frequency, and load-displacement relationships were determined. The results were compared with those obtained by static lateral loading tests on the same specimen by the authors in previous studies. Earthquake response analyses were carried out by applying a structural model proposed by the authors based on the results of static lateral loading tests. The results of the analysis and shaking table tests are compared, and the validity of the proposed structural model is discussed.

p53 Isoforms in Cellular Senescence- and Ageing-Associated Biological and Physiological Functions.

Cellular senescence, a term originally used to define the characteristics of normal human fibroblasts that reached their replicative limit, is an important factor for ageing, age-related diseases including cancer, and cell reprogramming. These outcomes are mediated by senescence-associated changes in gene expressions, which sometimes lead to the secretion of pro-inflammatory factors, or senescence-associated secretory phenotype (SASP) that contribute to paradoxical pro-tumorigenic effects. p53 functions as a transcription factor in cell-autonomous responses such as cell-cycle control, DNA repair, apoptosis, and cellular senescence, and also non-cell-autonomous responses to DNA damage by mediating the SASP function of immune system activation. The human TP53 gene encodes twelve protein isoforms, which provides an explanation for the pleiotropic p53 function on cellular senescence. Recent reports suggest that some short isoforms of p53 may modulate gene expressions in a full-length p53-dependent and -independent manner, in other words, some p53 isoforms cooperate with full-length p53, whereas others operate independently. This review summarizes our current knowledge about the biological activities and functions of p53 isoforms, especially Δ40p53, Δ133p53α, and p53ß, on cellular senescence, ageing, age-related disorder, reprogramming, and cancer. Numerous cellular and animal model studies indicate that an unbalance in p53 isoform expression in specific cell types causes age-related disorders such as cancer, premature ageing, and degenerative diseases.

A-674563 increases chondrocyte marker expression in cultured chondrocytes by inhibiting Sox9 degradation.

The implantation of autologous chondrocytes is a therapeutic treatment for articular cartilage damage. However, the benefits are limited due to the expansion of chondrocytes in monolayer culture, which causes loss of chondrocytic characters. Therefore, culture conditions that enhance chondrocytic characters are needed. We screened 5822 compounds and found that A-674563 enhanced the transcription of several chondrocyte marker genes, including Col2a1, Acan and Col11a2, in mouse primary chondrocytes. Experiments using cycloheximide, MG132 and bafilomycin A1 have revealed that Sox9 is degraded through the ubiquitin-proteasome pathway and that A-674563 inhibits this degradation, resulting in larger amount of Sox9 protein. RNA sequencing transcriptome analysis showed that A-674563 increases the expression of the gene that encodes ubiquitin-specific peptidase 29, which is known to induce the deubiquitination of proteins. Although the precise mechanism remains to be determined, our findings indicated that A-674563 could contribute to culture conditions that expand chondrocytes without losing chondrocytic characters.

UROLITH PREVALENCE AND RISK FACTORS IN ASIAN SMALL-CLAWED OTTERS ( AONYX CINEREUS).

Uroliths (urinary stones) are routinely found in both domestic and exotic animals kept under human care. In zoos, Asian small-clawed otters (ASCOs, Aonyx cinereus) have been identified as being particularly prone to this disease. Risk factors are thought to be nutritional; however, recommendations contradict each other, depending on which physiological model was used. Our study aimed to determine the prevalence of uroliths in ASCOs under human care and to evaluate which feeding patterns and nutrients may be linked to their occurrence. Questionnaires were sent to zoos holding ASCOs in North America (AZA), Europe (EAZA), and Japan and Southeast Asia (Asia) asking about diets and medical histories of all ASCOs alive or dead within the last 10 yr. A risk-factor style binary logistic regression was conducted on these data. A total of 94 questionnaires were received; however, only 56 were usable (15.6% return rate), representing 161 otter cases. AZA had the significantly highest incidence of kidney stones (62.8%), followed by EAZA (12.9%) and Asia (9.4%). Age and calcium were risk factors, whereas crude protein and sodium were protective. Therefore, calcium may need to be controlled within their diet. A diet high in fish and crustaceans may be beneficial and is consistent with wild ASCO diets. The feline model may be the best choice out of other models; however, many factors cannot be compared with ASCO, such as urinary pH.

Prevalence of small for gestational age (SGA) and short stature in children born SGA who qualify for growth hormone treatment at 3 years of age: Population-based study.

BACKGROUND: To treat children born small for gestational age (SGA) with severe short stature, treatment with growth hormone (GH) has been approved in the USA, Europe, and Japan, but no population-based studies have reported their prevalence. The aims of this study were to investigate the prevalence of SGA and short stature in children born SGA who qualify for GH treatment at 3 years of age in a Japanese population. METHODS: A population-based study was conducted in Kobe, Japan with 27 228 infants who were born between 2006 and 2008 and followed until 3 years of age. Prevalence of birthweight (BW) or birth length (BL) ≤ -2.0 standard deviation scores (SDS) for gestational age (GA; definition of SGA) was calculated. Short children born SGA who qualify for GH treatment at 3 years of age were estimated using the following criteria: BW and BL below the 10th percentile for GA, BW or BL ≤ -2.0 SDS for GA, and 2.5 SDS below the mean height for age. RESULTS: The prevalence of SGA was 3.5%. The estimated prevalence of short stature in children born SGA who met the criteria for GH treatment was 0.06%. The prevalence in infants born <34 weeks (0.39%) was significantly higher than that in infants born 34-41 weeks GA (0.05%, P = 0.02). CONCLUSIONS: The prevalence of SGA and short stature in children born SGA who qualify for GH treatment is approximately 1 of 30 infants and 1 of 1800 children, respectively. The risk is increased when children are born <34 weeks GA.

Fluorescence fingerprint as an instrumental assessment of the sensory quality of tomato juices.

BACKGROUND: Sensory analysis is an important standard for evaluating food products. However, as trained panelists and time are required for the process, the potential of using fluorescence fingerprint as a rapid instrumental method to approximate sensory characteristics was explored in this study. RESULTS: Thirty-five out of 44 descriptive sensory attributes were found to show a significant difference between samples (analysis of variance test). Principal component analysis revealed that principal component 1 could capture 73.84 and 75.28% variance for aroma category and combined flavor and taste category respectively. Fluorescence fingerprints of tomato juices consisted of two visible peaks at excitation/emission wavelengths of 290/350 and 315/425 nm and a long narrow emission peak at 680 nm. The 680 nm peak was only clearly observed in juices obtained from tomatoes cultivated to be eaten raw. The ability to predict overall sensory profiles was investigated by using principal component 1 as a regression target. Fluorescence fingerprint could predict principal component 1 of both aroma and combined flavor and taste with a coefficient of determination above 0.8. CONCLUSION: The results obtained in this study indicate the potential of using fluorescence fingerprint as an instrumental method for assessing sensory characteristics of tomato juices.

Method of determining the optimal dilution ratio for fluorescence fingerprint of food constituents.

Quantitative determination by fluorescence spectroscopy is possible because of the linear relationship between the intensity of emitted fluorescence and the fluorophore concentration. However, concentration quenching may cause the relationship to become nonlinear, and thus, the optimal dilution ratio has to be determined. In the case of fluorescence fingerprint (FF) measurement, fluorescence is measured under multiple wavelength conditions and a method of determining the optimal dilution ratio for multivariate data such as FFs has not been reported. In this study, the FFs of mixed solutions of tryptophan and epicatechin of different concentrations and composition ratios were measured. Principal component analysis was applied, and the resulting loading plots were found to contain useful information about each constituent. The optimal concentration ranges could be determined by identifying the linear region of the PC score plotted against total concentration.

[Stem cell aging and the implications for stem cell-based therapies for aging-related diseases and aged tissues].

Adult stem cells exist in most mammalian tissues to maintain their homeostasis and help repair them. Reductions in adult stem cell function and/or number are clearly associated with aging, however, the causal correlations between such findings and the effects of aging are largely unknown. Some stem cell functional changes, such as the loss of lineage specificity and self-renewal capacity, senescence and transformation, arise in stem cells autonomously during the aging process. These autonomous changes of stem cell functions reflect the damaging effects of age on the genome, epigenome, and proteome. Other stem cell functional changes are influenced by the age-related changes in the local microenvironments (niches) or systemic environments. If stem cell-based therapy can be used not only for age-related degenerative diseases, but also normal functional declines associated with aging, consideration of the behavior of stem cells based on effects from the local microenvironments (niches) and systemic environments in older individuals will therefore be needed.

Clinical Utility of SARS-CoV-2 Antibody Titer Multiplied by Binding Avidity of Receptor-Binding Domain (RBD) in Monitoring Protective Immunity and Clinical Severity.

Conventional serum antibody titer, which expresses antibody level, does not provide antigen binding avidity of the variable region of the antibody, which is essential for the defense response to infection. Here, we quantified anti-SARS-CoV-2 antibody binding avidity to the receptor-binding domain (RBD) by competitive binding-inhibition activity (IC50) between SARS-CoV-2 S1 antigen immobilized on the DCP microarray and various RBD doses added to serum and expressed as 1/IC50 nM. The binding avidity analyzed under equilibrium conditions of antigen-antibody binding reaction is different from the avidity index measured with the chaotropic agent, such as urea, under nonequilibrium and short-time conditions. Quantitative determination of the infection-protection potential of antibodies was assessed by ABAT (antigen binding avidity antibody titer), which was calculated by the quantity (level) × quality (binding avidity) of antibodies. The binding avidity correlated strongly (r = 0.811) with cell-based virus-neutralizing activity. Maturation of the protective antibody induced by repeated vaccinations or SARS-CoV-2 infection was classified into three categories of ABAT, such as an initial, low, and high ABAT. Antibody maturity correlated with the clinical severity of COVID-19. Once a mature high binding avidity was achieved, it was maintained for at least 6-8 months regardless of the subsequent change in the antibody levels.

Changes in the texture and viscoelastic properties of bread containing rice porridge during storage.

The objective of this study was to investigate the effects of rice porridge on the texture and viscoelastic properties of bread during storage. Three types of bread, wheat flour bread, 15% rice flour bread, and 15% rice porridge bread, were prepared. After baking and storing the bread for 24 h, 48 h, and 72 h at room temperature, we measured the texture and viscoelastic properties of the bread crumbs by texture profile analysis (TPA) and creep test. The 15% rice porridge bread showed a significantly higher specific volume and maintained softer crumbs than the other two types (p<0.05). It also had a slightly stickier texture than the others. It can be concluded that rice porridge improves the specific volume, texture, and viscoelastic properties of bread crumbs during storage.

Predicting the buckwheat flour ratio for commercial dried buckwheat noodles based on the fluorescence fingerprint.

A rapid method for predicting the buckwheat flour ratio of dried buckwheat noodles was developed by using the fluorescence fingerprint and partial least squares regression. Fitting the calibration model to validation datasets showed R(2)=0.78 and SEP=12.4%. The model was refined for a better fit by deleting several samples containing additional ingredients. The best fit was finally obtained (R(2)=0.84 and SEP=10.4%) by deleting the samples containing vinegar, green tea, seaweed, polysaccharide thickener, and yam. This result demonstrates that a calibration model with high accuracy could be constructed based on samples similar in material composition. The developed methodology requires no complex preprocessing, enables rapid measurement with a small sample amount, and would thus be suitable for practical application to the food industry.

Visualization of gluten and starch distributions in dough by fluorescence fingerprint imaging.

A novel method combining imaging techniques and fluorescence fingerprint (FF) data measurement was developed to visualize the distributions of gluten and starch in dough without any preprocessing. Fluorescence images of thin sections of gluten, starch, and dough were acquired under 63 different combinations of excitation and emission wavelengths, resulting in a set of data consisting of the FF data for each pixel. Cosine similarity values between the FF of each pixel in the dough and those of gluten and starch were calculated. Each pixel was colored according to the cosine similarity value to obtain a pseudo-color image showing the distributions of gluten and starch. The dough sample was then fluorescently stained for gluten and starch. The stained image showed patterns similar to the pseudo-color FF image, validating the effectiveness of the FF imaging method. The method proved to be a powerful visualization tool, applicable in fields other than food technology.

[Implementation and evaluation of a childhood dental health program in a community: twenty-year observational data].

UNLABELLED: PURPOSE To evaluate a long-term community-based childhood dental health program comprising of caries screenings of one- and two-year old kindergarten and pre-school children as well as in groups aged 1.5- and 3-years. METHODS: Fluoride treatment was introduced to 1.5-year, 2-year, 3-year screening from 1995, 1-year screening from 1997, and kindergarten/pre-school children from 2002 in Kyowa town, Makabe, Ibaraki (Kyowa district of Chikusei city) . Tests for caries prediction and caries bacteria load were also introduced from 1990 and 1995, respectively. To evaluate improvement in the dental health program between 1984 and 2004, we compared the prevalence trend for dental caries in Kyowa town with that in 7 other adjacent communities within the same public health center area, the Ibaraki prefectural average and data for all Japan. RESULTS: Between 1984 and 2004, the prevalence of dental caries declined by 59% for 3-year children with a large decrease from 1995, and by 57% for 1.5-year children. Decline during the twenty years ranked first and second among the communities, respectively, and was also larger than the prefectural and national averages. For 3-year children, the reduced prevalence of dental caries may have been enhanced by the introduction of fluoride treatment in 1995. CONCLUSION: A community-based dental health program including fluoride treatment for 1- and 2-year old kindergarten/pre-school children as well as for those aged 3- and 1.5 years may have had a larger impact on reducing dental caries in Kyowa town than in other communities, Ibaraki prefecture and all Japan.

Effect of potato flour on quality and staling properties of wheat-potato flour bread.

To elucidate the impact of potato flour (PF) on quality changes and staling characteristics of the composite bread from wheat-potato flour (WPF), the physicochemical (specific volume, colority, sensory value, texture, and viscosity) properties, and staling (X-ray diffraction and water migration) properties of bread were investigated. The quality of composite bread was comparable to wheat bread when addition level of PF at 20%, but decreased when the addition level increased to 30% or more, and became unacceptable at 50%. A chewy mouthfeel and an elastic and none-crumbly texture were observed on composite bread, which had higher hardness than wheat bread, and could keep on both longer linear distance and higher linear force during compression test. It indicated that such new parameters other than hardness should be introduced to coordinate with the texture quality of composite bread. During storage, the higher addition level of PF significantly decreased crystallinity of composite bread and slowed water migration rate from the crumb to crust, suggesting that PF had antistaling effect on composite bread, which was further emphasized by the fact that the setback value of the WPF decreased with the increase of PF addition.

Evaluation of FGFR inhibitor ASP5878 as a drug candidate for achondroplasia.

Achondroplasia is caused by gain-of-function mutations in FGFR3 gene and leads to short-limb dwarfism. A stabilized analogue of C-type natriuretic peptide (CNP) is known to elongate bone by interacting with FGFR3 signals and thus is a promising drug candidate. However, it needs daily administration by percutaneous injection. FGFR inhibitor compounds are other drug candidates for achondroplasia because they directly fix the mutant protein malfunction. Although FGFR inhibitors elongate the bone of model mice, their adverse effects are not well studied. In this study, we found that a new FGFR inhibitor, ASP5878, which was originally developed as an anti-cancer drug, elongated the bone of achondroplasia model male mice at the dose of 300 µg/kg, which confers an AUC of 275 ng·h/ml in juvenile mice. Although ASP5878 was less effective in bone elongation than a CNP analogue, it is advantageous in that ASP5878 can be administered orally. The AUC at which minimal adverse effects were observed (very slight atrophy of the corneal epithelium) was 459 ng·h/ml in juvenile rats. The positive discrepancy between AUCs that brought efficacy and minimal adverse effect suggests the applicability of ASP5878 to achondroplasia in the clinical setting. We also analyzed effects of ASP5878 in a patient-specific induced pluripotent stem cell (iPSC) model for achondroplasia and found the effects on patient chondrocyte equivalents. Nevertheless, cautious consideration is needed when referring to safety data obtained from its application to adult patients with cancer in clinical tests.

Diagnostic performance of serum interferon gamma, matrix metalloproteinases, and periostin measurements for pulmonary tuberculosis in Japanese patients with pneumonia.

Serum markers that differentiate between tuberculous and non-tuberculous pneumonia would be clinically useful. However, few serum markers have been investigated for their association with either disease. In this study, serum levels of interferon gamma (IFN-γ), matrix metalloproteinases 1 and 9 (MMP-1 and MMP-9, respectively), and periostin were compared between 40 pulmonary tuberculosis (PTB) and 28 non-tuberculous pneumonia (non-PTB) patients. Diagnostic performance was assessed by analysis of receiver-operating characteristic (ROC) curves and classification trees. Serum IFN-γ and MMP-1 levels were significantly higher and serum MMP-9 levels significantly lower in PTB than in non-PTB patients (p < 0.001, p = 0.002, p < 0.001, respectively). No significant difference was observed in serum periostin levels between groups. ROC curve analysis could not determine the appropriate cut-off value with high sensitivity and specificity; therefore, a classification tree method was applied. This method identified patients with limited infiltration into three groups with statistical significance (p = 0.01), and those with MMP-1 levels < 0.01 ng/mL and periostin levels ≥ 118.8 ng/mL included only non-PTB patients (95% confidence interval 0.0-41.0). Patients with extensive infiltration were also divided into three groups with statistical significance (p < 0.001), and those with MMP-9 levels < 3.009 ng/mL included only PTB patients (95% confidence interval 76.8-100.0). In conclusion, the novel classification tree developed using MMP-1, MMP-9, and periostin data distinguished PTB from non-PTB patients. Further studies are needed to validate our cut-off values and the overall clinical usefulness of these markers.