Precocious puberty in patients with Pompe disease.

Introduction: The life expectancy of Pompe disease patients has increased due to improved neonatal screening and enzyme replacement therapy. Nevertheless, the potential effect of frequent medical device exposure on pubertal development in these patients is not well understood, so further investigation is warranted. Methods: In this cross-sectional study, we assessed the growth and puberty of nine Pompe disease patients. In addition, to determine the effects of frequent plastic medical device exposure in these patients, we measured urinary phthalate metabolites before and one day after enzyme replacement therapy. Results: Five out of nine patients (55%) with Pompe disease on enzyme replacement therapy had precocious puberty. Patients with precocious puberty had significantly shorter predicted adult heights compared to those with normal puberty (p = 0.014). The levels of mono-2-ethylhexyl phthalate (MEHP) and mono(2-ethyl-5-carboxypentyl) phthalate (MECPP) increased after enzyme replacement therapy, but the average levels of phthalate metabolites did not significantly differ between patients with normal and precocious puberty. Conclusion: Pompe disease patients on enzyme replacement therapy tend to have precocious puberty, which may reduce their adult height. There are no significant differences in urinary phthalate metabolites between normal and precocious puberty patients. Regular follow-up of growth and puberty in Pompe disease patients is important to improve their health outcomes.

Serum brain-derived neurotrophic factor levels as a predictor for Alzheimer disease progression.

BACKGROUND: Brain-derived neurotrophic factor (BDNF) has been implicated in the pathophysiology of Alzheimer's disease (AD), and decreased peripheral levels of this protein are associated with an increased risk of developing the disease. This study focuses on whether serum BDNF levels could be used as a predictor of AD progression. METHODS: In this longitudinal observational study, we recruited cognition normal participants (N = 98) and AD (N = 442) from the Clinic at the Taipei Veterans General Hospital. We conducted a mini-mental status exam, a 12-item memory test, a categorical verbal fluency test, and a modified 15-item Boston naming test. A Serum BDNF level and apolipoprotein E ( APOE ) allele status were measured. The AD patients were followed prospectively. Based on the difference of MMSE scores, these patients were divided into fast decliners (decline ≥ 3/y) and slow decliners (MMSE decline < 3/y). Logistic regression was conducted to examine the impact of serum BDNF levels and other factor on the likelihood of AD patients being slow decliners. Pearson's correlation was used to estimate the relationship between serum BDNF levels and the score of neuropsychological tests. RESULTS: In a logistic regression model containing serum BDNF levels, age, sex, APOE4 carrier status, education levels, and baseline MMSE score, higher serum BDNF levels were associated with a slower rate of cognitive decline in the AD group. Serum BDNF levels positively correlated with the results of multiple neuropsychological tests. CONCLUSION: BDNF is a protective factor against AD progression and likely plays a role in establishing a link between AD pathology and clinical manifestations.

Curated incidence of lysosomal storage diseases from the Taiwan Biobank.

Lysosomal storage diseases (LSDs) are a group of metabolic disorders resulting from a deficiency in one of the lysosomal hydrolases. Most LSDs are inherited in an autosomal or X-linked recessive manner. As LSDs are rare, their true incidence in Taiwan remains unknown. In this study, we used high-coverage whole-genome sequencing data from 1,495 Taiwanese individuals obtained from the Taiwan Biobank. We found 3826 variants in 71 genes responsible for autosomal recessive LSDs. We first excluded benign variants by allele frequency and other criteria. As a result, 270 variants were considered disease-causing. We curated these variants using published guidelines from the American College of Medical Genetics and Genomics (ACMG). Our results revealed a combined incidence rate of 13 per 100,000 (conservative estimation by pathologic and likely pathogenic variants; 95% CI 6.92-22.23) to 94 per 100,000 (extended estimation by the inclusion of variants of unknown significance; 95% CI 75.96-115.03) among 71 autosomal recessive disease-associated genes. The conservative estimations were similar to those in published clinical data. No disease-causing mutations were found for 18 other diseases; thus, these diseases are likely extremely rare in Taiwan. The study results are important for designing screening and treatment methods for LSDs in Taiwan and demonstrate the importance of mutation curation to avoid overestimating disease incidences from genomic data.

Core vocabulary for AAC practice from Mandarin Chinese-speaking Taiwanese without disabilities.

The purpose of this study was to select a core vocabulary list obtained from Mandarin Chinese-speaking Taiwanese persons without disabilities. Mandarin Chinese is dominant and official language of Taiwan. A total of 28 participants, equally divided among seven age groups, were recruited for the study. In all, 112 samples across different communication contexts were collected. Results indicated that 100 core words selected had coverage of 66.7% of the entire composite sample. The proportion of function words versus content words in the top 100 core words was 11% and 89%, respectively. The core vocabulary was categorized into eight parts of speech, including nouns, pronouns, numbers, adverbs, determiners, prepositions, adjectives, and verbs. Implications, limitations, and further research are discussed.

Morphological changes of foveal cysts as a predictor for visual response to anti-vascular endothelial growth factor treatments in diabetic macular edema : Degenerative cyst in DME.

PURPOSE: To investigate morphological changes of intraretinal cyst in association with visual acuity following treatment for diabetic macular edema. METHODS: This retrospective study enrolled 105 eyes from 105 treatment naïve patients with diabetic macular edema following anti-vascular endothelial growth factor injections. Best-corrected visual acuity (BCVA) and optical coherence tomography (OCT) data were obtained at baseline, 1, 3, 6, and 12 months. The width and height of the largest intraretinal cyst (IRC) at all different visits were measured and were correlated to final visual acuity by receiver operating characteristic curve. The exudative feature was defined by the presence of hard exudates. Multivariate logistic regression was used to select the independent predictor for visual outcomes. RESULTS: Intraretinal cyst width but not the cyst height after treatment at 1 month independently predicted final visual loss of ten letters or more (multivariate P = 0.009). The optimal cutoff value was 196 um with a sensitivity of 0.889 and a specificity of 0.656. Eyes with large IRC width using this cutoff were consistently larger than those with small IRC width through 12 months (P = 0.008, Mann-Whitney U test). Small IRC width < 196 um at 1 month was more likely to coexist with exudative feature (P = 0.011, Fisher's exact test). Among baseline factors, large IRC width predicted IRC width ≥ 196 um at 1 month (multivariate P < 0.001). CONCLUSION: Cyst morphology following intravitreal injection predicts visual outcomes. Eyes with IRC width ≥ 196 um after treatment at 1 month tends to be more degenerative, and less likely to coexist with exudative feature.

Dyadic Conversation between Mandarin-Chinese-Speaking Healthy Older Adults: From Analyses of Conversation Turns and Speaking Roles.

Older adults' daily conversations with other older adults enable them to connect to their surrounding communities and improve their friendships. However, typical aging processes and fluctuations in family caring might cause conversation changes. The purpose of this study was to explore the quantitative contributions of conversation turns (CTs) and speaking roles (SRs) in Mandarin-Chinese-speaking conversation dyads between mutually familiar healthy older adults (HOAs). A total of 20 HOAs aged 65 or over were recruited. Each dyad conversed for ten minutes once a week for five weeks, five sessions per dyad, for a total of 50 sessions. The frequency and percentages of the coded CTs and SRs contributed by each HOA were individually tallied and calculated. Quantitatively symmetrical contributions of CTs and SRs occurred in Mandarin-Chinese-speaking conversation dyads between mutually familiar HOAs. Although typical aging processes might change conversations, both Mandarin-Chinese-speaking HOAs serve as active interlocutors to each other in taking CTs and SRs to co-construct their conversation processes and content in their dyadic conversation. Sufficient knowledge of conversation co-constructions might lead them to have more supportive environments to connect to surrounding communities and improve their friendships.

Adult height of children with congenital adrenal hyperplasia due to 21-hydroxylase deficiency.

BACKGROUND/PURPOSES: Congenital adrenal hyperplasia attributable to 21-hydroxylase deficiency (21-OHD) is a disorder of adrenal steroidogenesis. Achievement of optimal growth by such patients is challenging. We evaluated the adult height of Taiwanese children with 21-OHD and the effect of a gonadotropin-releasing hormone analogue (GnRHa) in patients with central precocious puberty (CPP) complicating 21-OHD. METHODS: Among 116 patients with 21-OHD in Taiwan, 90 who had attained adult height were subjected to an analysis of height outcomes. Nine with progressive CPP were treated with GnRHa and the effects of this therapy on adult height were further analyzed. RESULTS: In the pre-screening era, the percentage of boys with 21-OHD was lower than expected. Although neonatal screening can prevent mortality caused by adrenal crisis, some cases may be missed. The pooled mean adult height of the 78 patients treated with conventional therapy were -1.1 SD and -0.5 SD adjusting for the genetic potential. The disease features affecting height outcomes are the genetic height potential and in boys the simple virilizing type. Nine patients with CPP were treated with GnRHa in addition to conventional therapy; the mean adult height increased from the predicted -4.1 SD to -1.0 SD after 6.0 ± 2.5 years of treatment. CONCLUSION: Patients with 21-OHD had poorer mean adult height. A high caregiver's index of suspicion is required for the early diagnosis of patients with 21-OHD missed on neonatal screening. Adjuvant therapy with GnRHa can improve the adult height of patients with CPP complicating 21-OHD.

Using the ICF Framework to Assess Communicative Competence in Dyadic Communication among Children and Adolescents Who Use Augmentative and Alternative Communication Devices in Taiwan.

Augmentative and alternative communication (AAC) devices enable children and adolescents (CAD) with communication disorders to communicate competently and develop friendships through communicative competence (CC). Existing assessment tools are unable to indicate whether CAD aged 0 to 18 years would competently use the subsidized AAC devices provided by the Ministry of Health and Welfare in Taiwan. This study, thus, aimed to develop an assessment tool by using the International Classification of Functioning, Disability and Health (ICF) to measure CC in dyadic communication among CAD using AAC devices. Five speech-language pathologists (SLPs), five special education teachers, and four AAC experts (14 in total) selected codes relevant to the four domains of CC via the Delphi method. Next, they categorized the selected codes into one of the four domains of CC through a face-to-face expert panel. A total of 112 codes were listed in the tool and fully classified into the four domains of CC. Among these, seven codes were concurrently placed under two domains of CC. Consequently, this study developed an assessment tool by employing the ICF for children and youth core set using universal qualifiers to measure the relative levels of CC in dyadic communication among CAD who use AAC devices in their daily life.

A missense variant in the nuclear localization signal of DKC1 causes Hoyeraal-Hreidarsson syndrome.

Hoyeraal-Hreidarsson syndrome (HHS) is the most severe form of dyskeratosis congenita (DC) and is caused by mutations in genes involved in telomere maintenance. Here, we identified male siblings from a family with HHS carrying a hemizygous mutation (c.1345C > G, p.R449G), located in the C-terminal nuclear localization signal (NLS) of the DKC1 gene. These patients exhibit progressive cerebellar hypoplasia, recurrent infections, pancytopenia due to bone marrow failure, and short leukocyte telomere lengths. Single-cell RNA sequencing analysis suggested defects in the NLRP3 inflammasome in monocytes and the activation and maturation of NK cells and B cells. In experiments using induced pluripotent stem cells (iPSCs) from patients, DKC1_R449G iPSCs had short telomere lengths due to reduced levels of human telomerase RNA (hTR) and increased cytosolic proportions of DKC1. Treatment with dihydroquinolizinone RG7834 and 3'deoxyanosine cordycepin rescued telomere length in patient-derived iPSCs. Together, our findings not only provide new insights into immunodeficiency in DC patients but also provide treatment options for telomerase insufficiency disorders.

Development of ICF core set to profile communicative competence in dyadic communication among adults who use communication devices in Taiwan.

PURPOSE: This study aimed to develop an ICF core set to profile communicative competence in dyadic communication among adults who use communication devices in Taiwan. MATERIALS AND METHODS: The study was conducted following the release of a manual by the World Health Organisation (WHO) on how to develop the International Classification of Functioning, Disability and Health (ICF) core set in three phases: Preparatory Phase, Phase I, and Phase II. The Preparatory Phase helped collect and sort second-level ICF codes into a candidate list based on different perspectives, including the Systematic Literature Review Phase (i.e., researchers' perspectives), Empirical Multi-Center Study Phase (i.e., clinical perspectives), Qualitative Study Phase (i.e., perspectives of individuals with a health condition), and Expert Survey Phase (i.e., health professionals' perspectives). An ICF core set was developed from the candidate list in Phase I through the Delphi technique, and the content validity of this core set was assessed in Phase II. RESULTS: Altogether, 94 s-level ICF codes in the candidate list from the Preparatory Phase were included in the three rounds of the Delphi technique for Phase I. Finally, these 94 s-level ICF codes were validated and included in the core set for Phase II, including 28 in Body Functions (b), 0 in Body Structures (s), 42 in Activities and Participation (d), and 24 in Environmental Factors (e). CONCLUSIONS: The developed ICF core set provides an evaluation tool to profile communicative competence in dyadic communication among adults using communication devices. This core set identifies the gap and future opportunities for further examining the care providers' roles, together with exploring the environmental facilitators and barriers. The implications concerning rehabilitation, limitations, and the way forward are discussed.Implications to RehabilitationThis core set was first developed in the context of Taiwan from the perspectives of adults and professionals who used communication devices to profile communicative competence in dyadic communication.This core set, which can be utilised across health care disciplines, can serve as the foundation for more holistic evaluation, profiling levels of communicative competence in daily dyadic communication among adults who use communication devices.Rehabilitation providers may decide, based on this core set, if communication devices should be proposed for adults again in order to minimise the abandonment of subsidised communication devices.Levels of communicative competence in daily dyadic communication among adults who use communication devices can be profiled through this ICF core set.

Aggregation Control, Surface Passivation, and Optimization of Device Structure toward Near-Infrared Perovskite Quantum-Dot Light-Emitting Diodes with an EQE up to 15.4.

In recent years, the performance of perovskite quantum dots (QDs) and QD-based light-emitting diodes (QLEDs) has improved greatly, with electroluminescence (EL) efficiency of green and red emission exceeding 20%. However, the development of perovskite near-infrared (NIR) QLEDs has reached stagnation, where the reported maximum EL efficiency is still below 6%, limiting their further applications. In this work, new NIR-emissive FAPbI3 QDs are developed by post-treating long alkyl-encapsulated QDs with 2-phenylethylammonium iodide (PEAI). The incorporation of PEAI reduces the QD surface defects for giving a high photoluminescence quantum yield up to 61.6%. The n-octane solution of PEAI-passivated FAPbI3 QDs is spin coated on top of the PEDOT:PSS-treated ITO electrode modified with a thermally crosslinked hole-transporting layer to give a full-coverage, smooth, and dense QD film. Incorporating with an effective electron-transporting material, CN-T2T, which has deep lowest unoccupied molecular orbital and good electron mobility, the optimal device with EL λmax at 772 nm achieves an external quantum efficiency up to 15.4% at a current density of 0.54 mA cm-2 (2.6 V), which is the highest efficiency ever reported for perovskite-based NIR QLEDs. This study provides a facile strategy to prepare high-quality perovskite QD films suitable for highly efficient NIR QLED applications.

Cross-Camera External Validation for Artificial Intelligence Software in Diagnosis of Diabetic Retinopathy.

Aims: To investigate the applicability of deep learning image assessment software VeriSee DR to different color fundus cameras for the screening of diabetic retinopathy (DR). Methods: Color fundus images of diabetes patients taken with three different nonmydriatic fundus cameras, including 477 Topcon TRC-NW400, 459 Topcon TRC-NW8 series, and 471 Kowa nonmyd 8 series that were judged as "gradable" by one ophthalmologist were enrolled for validation. VeriSee DR was then used for the diagnosis of referable DR according to the International Clinical Diabetic Retinopathy Disease Severity Scale. Gradability, sensitivity, and specificity were calculated for each camera model. Results: All images (100%) from the three camera models were gradable for VeriSee DR. The sensitivity for diagnosing referable DR in the TRC-NW400, TRC-NW8, and non-myd 8 series was 89.3%, 94.6%, and 95.7%, respectively, while the specificity was 94.2%, 90.4%, and 89.3%, respectively. Neither the sensitivity nor the specificity differed significantly between these camera models and the original camera model used for VeriSee DR development (p = 0.40, p = 0.065, respectively). Conclusions: VeriSee DR was applicable to a variety of color fundus cameras with 100% agreement with ophthalmologists in terms of gradability and good sensitivity and specificity for the diagnosis of referable DR.

Short stature leads to a diagnosis of Jansen-de Vries syndrome in two unrelated Taiwanese girls: A case report and literature review.

Short stature and intellectual disability are two of the major components of many dysmorphic syndromes. Jansen-de Vries syndrome (JDVS) is a rare syndromic disorder that was discovered recently using next-generation sequencing. It is characterized by hypotonia, developmental delay, a dysmorphic face, short stature, and high pain threshold and is caused by the variants of the protein phosphatase magnesium-dependent 1D (PPM1D) gene. Here, we report the first two cases of PPM1D mutations in Taiwan; both had de novo variants in exon 6. Both presented with short stature, developmental delay, and dysmorphic faces. In addition to the characteristics listed above, syndactyly was noted in one. Genetic studies should be considered when approaching a patient with growth retardation, intellectual disability, and other major or minor dysmorphisms.

Conversation turns and speaking roles contributed by Mandarin Chinese dyadic conversations between adults who use speech-generating devices and adults who use natural speech.

Few studies have critically examined contributed conversation turns and speaking roles in dyadic Mandarin Chinese conversations between adults who use natural speech and adults who use SGDs. An analysis of conversation turns considers how conversation participants co-construct conversation processes, whereas an analysis of speaking roles considers how conversation participants co-construct conversation contents. The purpose of the current study was to explore the quantitative contributions of conversation turns and speaking roles in familiar Mandarin Chinese conversation dyads between adults who use SGDs and adults who use natural speech. Five dyads were recruited, and each dyad contained two participants; an adult using an SGD and an adult using natural speech. Each dyadic conversation was individually video-recorded for about 20 minutes, six times. A total of 30 dyadic conversations were analyzed. The means and percentages of the coded conversation turns and speaking roles were computed. Quantitatively asymmetrical contributions of conversation turns and speaking roles occurred in the Mandarin Chinese conversation dyads between the adults who use SGDs and the adults who use natural speech. The adults who use natural speech contributed significantly more conversation turns and speaking roles than did the adults who use SGDs. Implications, limitations, and further research studies were discussed.

Patterns of ellipsoid zone change associated with visual outcome for diabetic macular oedema.

Clinical relevance: Optical coherence tomography biomarkers are dynamic and possibly change over time. The dynamic biomarkers may better predict improvement of final vision than those at baseline for diabetic macular oedema after treatment.Background: To investigate predictors for ellipsoid zone (EZ) dynamics and the association with visual acuity after treatment for diabetic macular oedemaMethods: A total of 135 eyes from 135 patients with anti-vascular endothelial growth factor treatment for diabetic macular oedema were retrospectively enrolled in this study. EZ status was evaluated by optical coherence tomography at baseline, 1, 3, 6, and 12 months post-operatively. Macular perfusion status was analysed by fluorescein angiography. Duration of EZ disruption was quantified by giving scores from 0 to 5 based on the sum of visits with evidence of EZ disruption. Linear regression models were used to predict factors for scores of EZ disruption. Binary logistic regression was used to predict various EZ biomarkers for final visual acuity improvement.Results: Thirty of 87 eyes (34.4%) with intact EZ at baseline developed subsequent EZ disruption. For those with EZ disruption at baseline, 17 of 48 (35.4%) patients had restoration of disrupted EZ over time. Non-perfused macula was associated with both earlier development of EZ disruption and poor restoration (p < 0.001 and p = 0.011, respectively). Non-perfused macula, absence of epiretinal membrane, and EZ disruption at baseline predicted higher EZ scores (p < 0.001, p = 0.022 and p < 0.001, respectively). Higher EZ scores (p = 0.016), but not baseline EZ disruption (p = 0.56), were less likely to have improvement of final vision 5 letters or more.Conclusion: Macular perfusion status played an independent role in EZ dynamics. Duration of EZ disruption could be more predictive than baseline status for improvement of vision over 12 months.

Intravitreal Aflibercept versus Ranibizumab for Diabetic Macular Edema in a Taiwanese Health Service Setting.

Purpose: To compare the visual and anatomical outcomes between intravitreal aflibercept and ranibizumab for diabetic macular edemaMethods: A total of 194 eyes from 194 patients (aflibercept n = 95, ranibizumab n = 99) were retrospectively enrolled in the study. All eyes fulfilled the key criteria including a baseline best-corrected visual acuity (BCVA) between 20 and 70 ETDRS letters, a central subfield thickness (CST) 300 µm or more. Primary outcomes were BCVA and CST at 1, 3, 6, and 12 months. Maintenance of vision was defined as visual loss of less than 5 letters over 6 to 12 months. Predictors for final visual acuity and visual maintenance were analyzed using multivariate regression models.Results: Both agents achieved comparable visual and anatomical outcomes at any time point over the course of follow-up (all p > .05). At 12 months, aflibercept group had higher proportions of visual gains 5, 10 and 15 letters or more (p = .014, p = .011, and p = .034, respectively). The mean number of injections was 5.0 ± 1.9 in ranibizumab group and 4.5 ± 1.9 in aflibercept group (p = .09). Ranibizumab predicted poor maintenance of vision (p = .009), but not the final visual acuity (univariate p = .1). Ranibizumab was more likely to have recurrence of subretinal fluid than aflibercept in 12 months after resolution of subretinal fluid at baseline (p = .016). Both aflibercept and ranibizumab had similar rates of loss to follow-up (p = .47) and occurrence of vitreous hemorrhage (p = .21).Conclusion: While both agents improved vision with resolution of edema, aflibercept maintained vision more effectively with less recurrence of subretinal fluid at 12 months in real-world settings.

Assessing Physiological Stress Responses to Word Retrieval in Individuals with Aphasia: A Preliminary Study.

OBJECTIVE: Word retrieval difficulty can be distressing for individuals with aphasia. However, physiological stress responses to word retrieval are relatively unstudied. This study aims to measure the physiological response (i.e., heart rate and respiratory rate) and perceived stress in individuals with aphasia during a naming task to understand whether word retrieval may be a stressor to them. METHOD: An aphasia group (n = 7) and a healthy group (n = 38) participated in 4 experimental conditions, including rest, counting, and high-frequency and low-frequency word conditions, while their physiological parameters were measured throughout the sessions. Stress ratings were also obtained for each condition. RESULTS: Significant differences in heart rate were found while participants were engaged in speech and/or language tasks, compared to the rest condition, for both groups. Significant differences in respiratory rate were found between the rest and low-frequency word conditions in the aphasia group. No significant difference was found in heart rate between 2 groups, but a difference in respiratory rate was found between 2 groups in the low-frequency word condition. There were some discrepancies between stress level and physiological parameters. CONCLUSIONS: Individuals with aphasia perceived greater stress in the naming tasks, however, the perceived stress was not necessarily parallel to the physiological parameters.

Association of Body Fluid Expansion With Optical Coherence Tomography Measurements in Diabetic Retinopathy and Diabetic Macular Edema.

Purpose: To evaluate associations of body fluid status with optical coherence tomography measurements in patients with diabetic retinopathy (DR). Methods: This prospective, cross-sectional study enrolled a total of 104 eyes from 104 patients with diabetes mellitus for fundus evaluations of DR and optical coherence tomography examinations. DR severity was graded via fundus photography. Systemic body fluid status was recorded via a body composition monitor with output values of total body water, extracellular water (ECW), intracellular water, and overhydration (OH). Relative overhydration (ROH) was defined as OH/ECW. Volume overload was defined as ROH ≥7%. Correlations of central subfield thickness (CST) with body fluid status were analyzed by partial correlation with adjustment for age, sex, and body mass index (BMI). Logistic regression analysis was used to evaluate factors associated with diabetic macular edema (DME). Results: Higher levels of ECW, OH, and ROH were correlated with thick CST in patients with DR (P = 0.006, 0.021, and 0.008, respectively), but not in those without any DR (all P > 0.05), after adjusting for age, sex, and BMI. Patients with DME (n = 31) had higher OH than DR patients without DME (n = 28) or those without any DR (n = 45) (P = 0.002 and P < 0.001, respectively). Multiple regression model showed that volume overload was the independent factor for the presence of DME (odds ratio, 9.532; 95% confidence interval, 2.898-31.348; P < 0.001). Conclusions: While both ECW and OH reflect CST in patients with DR, overhydration had particularly strong associations with DME. This study provides a novel insight into our current understanding regarding the pathogenesis for DME.

Dengue and Zika Virus Domain III-Flagellin Fusion and Glycan-Masking E Antigen for Prime-Boost Immunization.

The viral E proteins of dengue virus (DENV) and Zika virus (ZIKV) are the major viral proteins involved in receptor binding and fusion, and for the induction of protective antibodies against viral infections. DIII of the E proteins is an independent domain and stretches out on the virion surface that can elicit type-specific neutralizing antibodies. For recombinant DIII vaccine development, prime-boost immunizations can provide an advantage of eliciting more type-specific neutralizing antibodies by recalling DIII antigens after DIII booster to improve protection. Methods: The DIII of the E genes of DENV and ZIKV were fused with bacterial fliC gene for the expression of flagellin-DIII (FliC-DIII) fusion proteins. Prime-boost immunization strategies by the second-dose booster of four DENV serotype or ZIKV FliC-DIII fusion proteins were used to investigate the induction of neutralizing antibodies and protection against viral infections. Cross-reactive non-neutralizing antibodies in each group of antisera were also examined using in vitro antibody-dependent enhancement (ADE) assay. A series of glycan-masking E antigens were finally constructed for prime-boost immunizations to abolish the elicitation of cross-reactive non-neutralizing antibodies for ADE activity. Results: We showed that inclusion of a bivalent live-attenuated vaccine with a FliC-DIII booster is superior in eliciting neutralization titers and protection in vivo against all four-serotype DENVs. We also demonstrated that recombinant adenovirus vectors encoding four-serotype DENV prMEs with a FliC-DIII prime-boost scheme is capable of eliciting good antibody responses. In contract, recombinant adenovirus vector of ZIKV prME gene priming, followed by ZIKV FliC-DIII booster did not improve vaccine efficacy. The glycan-masking mutation on the ZIKV E protein ij loop (E-248NHT), but not on DENV2 E protein ij loop (E-242NHT), resulted in abolishing the elicitation of cross-reactive antibodies for DENV and ZIKV infection enhancements. Conclusions: Our findings can provide useful information for designing novel immunogens and vaccination strategies in an attempt to develop a safe and efficacious DENV or ZIKV vaccine.

Crystal structure of the programmed cell death 5 protein from Sulfolobus solfataricus.

Programmed cell death 5 (PDCD5) is a vital signaling protein in the apoptosis pathway in eukaryotes. It is known that there are two dissociated N-terminal regions and a triple-helix core in eukaryotic PDCD5. Structural and functional studies of PDCD5 from hyperthermophilic archaea have been limited to date. Here, the PDCD5 homolog Sso0352 (SsoPDCD5) was identified in Sulfolobus solfataricus, the SsoPDCD5 protein was expressed and crystallized, and the phase was identified by single-wavelength anomalous diffraction. The native SsoPDCD5 crystal belonged to space group C2 and diffracted to 1.49 Šresolution. This is the first crystal structure of a PDCD5 homolog to be solved. SsoPDCD5 shares a similar triple-helix bundle with eukaryotic PDCD5 but has a long α-helix in the N-terminus. A structural search and biochemical data suggest that SsoPDCD5 may function as a DNA-binding protein.