Volatomics-assisted characterization of aroma and off-flavor contributors in fresh and thermally treated kiwifruit juice.

The distinctive aroma profile of kiwifruit juice was significantly changed during thermal treatment, however, the theoretical basis for clarifying and controlling the changes was deficient. In this study, we applied volatomics techniques to investigate the contributors of off-flavors in thermally treated kiwifruit juice. Sixteen aroma compounds were identified to be responsible for the typical "fruity", "grassy", and "cucumber-like" flavors of fresh kiwifruit by two different fused silica capillary columns coupled with chromatography-olfactometry/detection frequency (GC-O/DF) analysis and calculation of odor activity value (OAV). Thirty-one odor-active compounds were determined as important contributors to the sensory profile of thermally treated kiwifruit juice, 14 of which were common to all varieties investigated. The key aroma compounds on fresh kiwifruit significantly decreased after thermal treatment, while decanal, (E)-2-decenal, methional, ß-damascenone, 1-octen-3-one, DMHF, and dimethyl sulfide which presented undesirable cooked cabbage/potato, roasted fruit, and sulfurous odors, were accumulated in a large amount. By applying PLSR analysis, (E)-2-decenal, methional, ß-damascenone, DMHF, and dimethyl sulfide were further verified to have great contributions to the formation of the cooked off-flavor during thermal treatment. Moreover, XX was found to be more thermal-sensitive and more prone to forming cooked off-flavors after thermal treatment. This study could provide theoretical guidance for the regulation of thermal-induced off-flavors during the manufacturing of kiwifruit juice.

Atypical prosodic realization by Mandarin-speaking autistic children: Evidence from tone sandhi and neutral tone.

Introduction Atypical prosodic features have been widely reported in autism spectrum disorder (ASD), primarily in non-tonal language speakers. Nevertheless, the prosodic realizations in autistic people who speak tonal languages were relatively understudied. This study aimed to investigate the acoustic and phonetic patterns at the word-level speech in Mandarin-speaking autistic and typically developing (TD) children at different age ranges. Methods Thirty Mandarin-speaking autistic children (15 three- to five-year-olds and 15 six- to eight-year-olds) were recruited into the ASD group. The TD group consisted of 30 age- and gender-matched children. We employed a picture-naming task to elicit the spontaneous speech production of Mandarin disyllabic words in which tone change processes occur, namely Tone 3 (T3) sandhi and neutral tone (T0). Results The phonetic analysis showed that the ASD group generally could produce typical-like T3 sandhi and T0 in terms of pitch height. However, relative to the TD group, they exhibited flatter pitch contours during T3 sandhi production. Moreover, the acoustic pitch mean of citation tones in the ASD group was also significantly higher, accompanied by more rigid pitch curves in contour tones. In addition, the atypical temporal realization in the ASD group was manifested by the longer duration of T0 and the earlier inflection position of T3. Conclusions Mandarin-speaking autistic children under eight had the phonological ability to produce context-dependent tones based on connected tonal information at the word level. Nevertheless, their phonetic prosodic realization of tone change processes was atypical. Our findings provide evidence of atypical prosody in autistic children who speak tone languages. Clinically, these findings may be attributable to underlying neural differences in autistic children.

Atypical patterns of tone production in tone-language-speaking children with autism.

Speakers with autism spectrum disorder (ASD) are found to exhibit atypical pitch patterns in speech production. However, little is known about the production of lexical tones (T1, T2, T3, T4) as well as neutral tones (T1N, T2N, T3N, T4N) by tone-language speakers with ASD. Thus, this study investigated the height and shape of tones produced by Mandarin-speaking children with ASD and their age-matched typically developing (TD) peers. A pronunciation experiment was conducted in which the participants were asked to produce reduplicated nouns. The findings from the acoustic analyses showed that although ASD children generally produced both lexical tones and neutral tones with distinct tonal contours, there were significant differences between the ASD and TD groups for tone height and shape for T1/T1N, T3/T3N, and T4/T4N. However, we did not find any difference in T2/T2N. These data implied that the atypical acoustic pattern in the ASD group could be partially due to the suppression of the F0 range. Moreover, we found that ASD children tended to produce more errors for T2/T2N, T3/T3N than for T1/T1N, T4/T4N. The pattern of tone errors could be explained by the acquisition principle of pitch, similarities among different tones, and tone sandhi. We thus concluded that deficits in pitch processing could be responsible for the atypical tone pattern of ASD children, and speculated that the atypical tonal contours might also be due to imitation deficits. The present findings may eventually help enhance the comprehensive understanding of the representation of atypical pitch patterns in ASD across languages.

The Incidence and Characteristics of Perinatal Stroke in Beijing: A Multicenter Study.

Objective: To assess the incidence, risk factors, and clinical characteristics of perinatal stroke in Beijing. Methods: This multicenter prospective study included all the live births from 17 representative maternal delivery hospitals in Beijing from March 1, 2019 to February 29, 2020. Neonates with a stroke were assigned to the study group. Clinical data, including general information, clinical manifestations, and risk factors, were collected. Up until 18 months after birth, neonates were routinely assessed according to the Ages and Stages Questionnaire (ASQ) and/or the Bayley scale. Statistical analysis was done using the chi-squared, t-tests, and logistic regression analysis using SPSS version 26.0. Outcomes: In total, 27 cases were identified and the incidence of perinatal stroke in Beijing was 1/2,660 live births, including 1/5,985 for ischemic stroke and 1/4,788 for hemorrhagic stroke. Seventeen cases (62.96%) of acute symptomatic stroke and convulsions within 72 h (10 cases, 37.04%) were the most common presentations. Ten patients showed no neurological symptoms and were found to have had a stroke through routine cranial ultrasonography after being hospitalized for non-neurological diseases. The risk factors include primiparity, placental or uterine abruption/acute chorioamnionitis, intrauterine distress, asphyxia, and severe infection. In the study group, 11.1% (3/27) of patients had adverse neurodevelopmental outcomes. The patients in the study group had lower scores for the ASQ than those in the control group in the communication, gross, and fine motor dimensions. Conclusion: The incidence of perinatal stroke in Beijing was consistent with that in other countries. Routine neuroimaging of infants with risk factors may enable identification of asymptomatic strokes in more patients. Patients who have suffered from a stroke may have neurological sequelae; therefore, early detection, treatment, and regular follow-ups are beneficial for improving their recovery outcomes.

Auditory-Motor Mapping Training Facilitates Speech and Word Learning in Tone Language-Speaking Children With Autism: An Early Efficacy Study.

PURPOSE: It has been reported that tone language-speaking children with autism demonstrate speech-specific lexical tone processing difficulty, although they have intact or even better-than-normal processing of nonspeech/melodic pitch analogues. In this early efficacy study, we evaluated the therapeutic potential of Auditory-Motor Mapping Training (AMMT) in facilitating speech and word output for Mandarin-speaking nonverbal and low-verbal children with autism, in comparison with a matched non-AMMT-based control treatment. METHOD: Fifteen Mandarin-speaking nonverbal and low-verbal children with autism spectrum disorder participated and completed all the AMMT-based treatment sessions by intoning (singing) and tapping the target words delivered via an app, whereas another 15 participants received control treatment. Generalized linear mixed-effects models were created to evaluate speech production accuracy and word production intelligibility across different groups and conditions. RESULTS: Results showed that the AMMT-based treatment provided a more effective training approach in accelerating the rate of speech (especially lexical tone) and word learning in the trained items. More importantly, the enhanced training efficacy on lexical tone acquisition remained at 2 weeks after therapy and generalized to untrained tones that were not practiced. Furthermore, the low-verbal participants showed higher improvement compared to the nonverbal participants. CONCLUSIONS: These data provide the first empirical evidence for adopting the AMMT-based training to facilitate speech and word learning in Mandarin-speaking nonverbal and low-verbal children with autism. This early efficacy study holds promise for improving lexical tone production in Mandarin-speaking children with autism but should be further replicated in larger scale randomized studies. Supplemental Material https://doi.org/10.23641/asha.16834627.

A genome-wide association study identifies FSHR rs2300441 associated with follicle-stimulating hormone levels.

Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) play critical roles in female reproduction, while the underlying genetic basis is poorly understood. Genome-wide association studies (GWASs) of FSH and LH levels were conducted in 2590 Chinese females including 1882 polycystic ovary syndrome (PCOS) cases and 708 controls. GWAS for FSH level identified multiple variants at FSHR showing genome-wide significance with the top variant (rs2300441) located in the intron of FSHR. The A allele of rs2300441 led to a reduced level of FSH in the PCOS group (ß = -.43, P = 6.70 × 10-14 ) as well as in the control group (ß = -.35, P = 6.52 × 10-4 ). In the combined sample, this association was enhanced after adjusting for the PCOS status (before: ß = -.38, P = 1.77 × 10-13 ; after: ß = -.42, P = 3.33 × 10-16 ), suggesting the genetic effect is independent of the PCOS status. The rs2300441 explained sevenfold higher proportion of the FSH variance than the total variance explained by the two previously reported FSHR missense variants (rs2300441 R2 = 1.40% vs rs6166 R2 = 0.17%, rs6165 R2 = 0.03%). GWAS for LH did not identify any genome-wide significant associations. In conclusion, we identified genome-wide significant association between variants in FSHR and circulating FSH first, with the top associated variant rs2300441 might be a primary contributor at the population level.

Quantitative proteomics analysis reveals alterations of lysine acetylation in mouse testis in response to heat shock and X-ray exposure.

Environmental stresses are important factors causing male infertility which attracts broad attention. Protein acetylation is a pivotal post-translational modification and modulates diverse physiological processes including spermatogenesis. In this study, we employed quantitative proteomic techniques and bioinformatics tools to analyze the alterations of acetylome profile of mouse testis after heat shock and X-irradiation. Overall, we identified 1139 lysine acetylation sites in 587 proteins in which 1020 lysine acetylation sites were quantified. The Gene Ontology analysis showed that the major acetylated protein groups were involved in generation of precursor metabolites and metabolic processes, and were localized predominantly in cytosolic and mitochondrial. Compared to the control group, 36 sites of 28 acetylated proteins have changed after heat shock, and 49 sites of 43 acetylated proteins for X-ray exposure. Some of the differentially acetylated proteins have been reported to be associated with the progression of spermatogenesis and male fertility. We observed the up-regulated acetylation level change on testis specific histone 2B and heat shock protein upon heat treatment and a sharp decline of acetylation level on histone H2AX under X-ray treatment, suggesting their roles in male germ cells. Notably, the acetylation level on K279 of histone acetyltransferase (Kat7) was down-regulated in both heat and X-ray treatments, indicating that K279 may be a key acetylated site and affect its functions in spermatogenesis. Our results reveal that protein acetylation might add another layer of complexity to the regulation for spermatogenesis, and further functional studies of these proteins will help us elucidate the mechanisms of abnormal spermatogenesis.

Transcriptome analysis of the responses to methyl methanesulfonate treatment in mouse pachytene spermatocytes and round spermatids.

Spermatogenesis is threatened by DNA alkylating agents, one major category of DNA damaging agents. Currently, little is known about the alterations in transcriptome profiling of the mouse spermatogenic cells in response to DNA alkylation at distinct stages of spermatogenesis. In this study, RNA sequencing (RNA-seq) was performed in pachytene spermatocytes (PS) and round spermatids (RS) at 0 or 30min following Methyl Methanesulfonate (MMS) treatment and with untreated controls. A large number of differentially expressed genes (DEGs) were identified by comparison of the three groups in PS and RS, respectively. Functional analyses of all DEGs highlighted the protein ubiquitination pathway and DNA damage response (DDR) network being the two main biological processes in common in the two cell types. Further analyses of the DEGs with 2-fold or more changes between 30min repair and control group indicated that several cytokine signaling pathways were the most strongly affected in PS and DDR related pathways in RS, respectively. Gene ontology (GO) analyses directly showed differential biological process (BP) affected between PS and RS, with "regulation of transcription" being most overrepresented in PS and "cellular response to stress" in RS, respectively. Moreover, 374 DDR-related genes in PS and 158 in RS among all DEGs were filtered and clustered, which showed dynamic expression patterns in PS and RS. Our analyses provide a transcriptional landscape for male germ cells in response to MMS during spermatogenesis.

Germline deletion of huntingtin causes male infertility and arrested spermiogenesis in mice.

Human Huntingtin (HTT), a Huntington's disease gene, is highly expressed in the mammalian brain and testis. Simultaneous knockout of mouse Huntingtin (Htt) in brain and testis impairs male fertility, providing evidence for a link between Htt and spermatogenesis; however, the underlying mechanism remains unclear. To understand better the function of Htt in spermatogenesis, we restricted the genetic deletion specifically to the germ cells using the Cre/loxP site-specific recombination strategy and found that the resulting mice manifested smaller testes, azoospermia and complete male infertility. Meiotic chromosome spread experiments showed that the process of meiosis was normal in the absence of Htt. Notably, we found that Htt-deficient round spermatids did not progress beyond step 3 during the post-meiotic phase, when round spermatids differentiate into mature spermatozoa. Using an iTRAQ-based quantitative proteomic assay, we found that knockout of Htt significantly altered the testis protein profile. The differentially expressed proteins exhibited a remarkable enrichment for proteins involved in translation regulation and DNA packaging, suggesting that Htt might play a role in spermatogenesis by regulating translation and DNA packaging in the testis.

Knockdown of p66Shc by siRNA injection rescues arsenite-induced developmental retardation in mouse preimplantation embryos.

Two-cell arrest plays a principal role in the elevated levels of embryo loss during the first week of development in mouse. Previously, we have shown that arsenic can apparently induce 2-cell arrest in mouse preimplantation embryo and the expression of oxidative stress adaptor protein p66(Shc) is up-regulated in this process. In the present study, we demonstrated that microinjection of p66(Shc) siRNA into the pronucleus of zygotes resulted in a markedly decrease in both mRNA and protein levels of p66(Shc). The arsenite-induced 2-cell arrests, along with a reduction in the levels of reactive oxygen species (ROS), were significantly inhibited and the number of embryos developing to morula stage concurrently increased upon p66(shc) siRNA microinjection. These findings indicate that knockdown of p66(shc) improves the developmental competence of arsenite-exposed embryos in vitro by increasing the resistance to oxidative stress. In addition, we highlight the utility of single-embryo analysis in preimplantation embryos.

[Distribution of arsenic species and its DNA damage in subchronic arsenite-exposed mice].

OBJECTIVE: To study the arsenic distribution, speciation, its effects on the balance of other elements and the DNA damage by subchronic arsenite exposure in mice. METHODS: The 8-week-old C57BL/6N mice were matched by weight and divided into control group and supplementation group, which were given 0 or 10 microg/ml of sodium arsenite in the drinking water, and continuous exposed for 6 months. RESULTS: Arsenic was found in various tissues and organs. The highest ones were in the kidney, lung and liver, reached (563.9 +/- 222.5), (458.6 +/- 191.0) and (279.8 +/- 81.2) ng/g, respectively while the lowest in the blood and brain, reached (82.2 +/- 26.7) ng/ml and (101.8 +/- 30.1) ng/g, respectively. Arsenic exists mainly in the form of dimethylarsinous acid (DMA). Compared to the control group, there was a significant difference (P < 0.05) between arsenic and chromium, copper, zinc, selenium, lead in some organs of arsenic exposed group, but not cadmium. Furthermore, the 8-hydroxydeoxyguanosine (8-OHdG) level of the exposed group was (149.1 +/- 1.0) ng/ml, which was significantly higher than the control group of (76.4 +/- 27.9) ng/ml. CONCLUSION: Arsenic accumulated in various tissues and organs mainly in the form of DMA, which affected the balance of chromium, copper, zinc, selenium and lead in the body, and led to DNA damage after subchronic exposure.

Role of spastin and protrudin in neurite outgrowth.

Hereditary spastic paraplegia (HSP) is a neurodegenerative disorder characterized by retrograde axonal degeneration that primarily affects long spinal neurons. The gene encoding spastin has a well-established association with HSP, and protrudin is a known binding partner of spastin. Here, we demonstrate that the N-terminal domain of protrudin mediates the interaction with spastin, which is responsible for neurite outgrowth. We show that spastin promotes protrudin-dependent neurite outgrowth in PC12 cells. To further confirm these physiological functions in vivo, we microinjected zebrafish embryos with various protrudin/spastin mRNA and morpholinos. The results suggest that the spinal cord motor neuron axon outgrowth of zebrafish is regulated by the interaction between spastin and protrudin. In addition, the putative HSP-associated protrudinG191V mutation was shown to alter the subcellular distribution and impair the yolk sac extension of zebrafish, but without significant defects in neurite outgrowth both in PC12 cells and zebrafish. Taken together, our findings indicate that protrudin interacts with spastin and induces axon formation through its N-terminal domain. Moreover, protrudin and spastin may work together to play an indispensable role in motor axon outgrowth.

Two novel HAND1 mutations in Chinese patients with ventricular septal defect.

BACKGROUND: The HAND1 gene encodes a basic helix-loop-helix (bHLH) transcription factor which plays an essential role in the development of heart. Mutations in HAND1 have been identified in congenital heart disease (CHD) patients with hypoplastic hearts and septal defects. The spectrum of CHD relating to HAND1 mutations needs further study. METHODS AND RESULTS: We screened HAND1 coding regions for mutations in 498 Chinese patients with CHD and 250 control subjects. We identified two novel non-synonymous mutations, c.217G>A (p.Gly73Ser) and c.456G>T (p.Lys152Asn), in the patients with ventricular septal defect (VSD). The two mutations were located in HAND1 evolutionarily conserved residues and enhanced the capability of HAND1 to form homodimers. CONCLUSIONS: This is the first report of mutations in the HAND1 gene in Chinese patients with VSD and provides new insight into the etiology of VSD.

CFC1 mutations in Chinese children with congenital heart disease.

BACKGROUND: Congenital heart disease (CHD) is one of the most common human birth defects. Over the last few decades, a variety of CHD-causing gene mutations have been identified. The aim of this study was to identify potential pathological mutations in the Cryptic (CFC1) gene in 500 Chinese children with CHD and to gain insight into the etiology of CHD. METHOD: Sequence analysis of CFC1 in 500 non-syndromic CHD patients and 250 healthy controls. RESULT: We identified three novel non-synonymous variants (c. G506T p. Gly169Val; c. G517A p. Gly173Arg; c. G658T p. Leu219Phe). These variants were not observed in 250 controls. All of the non-synonymous variants were located in or very close to the hydrophobic C-terminus. CONCLUSIONS: To our knowledge, this is the first study to suggest that CFC1 may be involved in the etiology of non-syndromic CHD in a Chinese population.

Teratocarcinoma-derived growth factor 1 (TDGF1) sequence variants in patients with congenital heart defect.

The goal of our study was to identify potential pathogenic mutations in the TDGF1 gene in Chinese people with isolated CHD, particularly those with VSD, and to provide further insight into the etiology of CHD. A total of 500 CHD Chinese patients were investigated for mutations in the TDGF1 gene. Thirteen variants were found among the 500 isolated VSD patients and 250 controls, including one non-synonymous variant identified in patients but not in controls. This work firstly provides human genetic evidence of TDGF1 involved in the pathogenesis of VSD, expanding our knowledge of the causative mutations of congenital heart defects, in particular, the causative mutations of VSD.

Interleukin 1 beta (IL-1ß) promoter C [-511] T polymorphism but not C [+3953] T polymorphism is associated with polycystic ovary syndrome.

PCOS is a complex multifactorial disorder involving a number of genetic and environmental factors. One of the genetic factors that has been associated with PCOS is Interleukin 1 beta (IL-1ß), is an important inflammatory cytokine that plays a regulatory role in both the body's immune and the inflammatory responses. All these responses appear to be are also affected in at least some women with PCOS. To investigate the possible association of polymorphisms of the IL-1ß gene with the occurrence and clinical characteristics of PCOS, we evaluated two common polymorphisms of the IL-1ß gene (promoter C [-511] T and exon 5 position [+3953]) in 200 Chinese women with PCOS and 177 healthy Chinese controls. We found the frequency of IL-1ß C/C [-511] genotype in PCOS was significantly higher than that in the controls (χ(2) = 15.48, df = 1, P < 0.001 OR = 2.73 95% CI: 1.64-4.56 by genotype; χ(2) = 10.21, df = 1, P = 0.001 by allele). However in contrast, no association between genotype and relative allele frequencies was observed for the C [+3953] T polymorphism for Chinese women with PCOS when compared to that for a similar group of Chinese women without PCOS (P = 0.35).

Identification of novel missense mutations of GDF9 in Chinese women with polycystic ovary syndrome.

The gene for growth differentiation factor 9 (GDF9) is expressed in human oocytes and has an important function in regulating early follicle growth and fertility. Polycystic ovary syndrome (PCOS) is one of the common defects that causes ovary dysfunction and is linked to aberrant processes in folliculogenesis. Previous studies have discovered several mutations in the screening of GDF9 in premature ovarian failure but none in PCOS. This current study focused on the mutational analysis of the coding region of GDF9 among 216 Chinese PCOS patients. Of the 10 different variants found in this study, five novel missense mutations in GDF9 were discovered namely c.15C>G, c.118T>G, c.133A>G, c.1025A>T and c.1275C>A. The above-mentioned mutations indicate GDF9 may be potentially associated with PCOS patients. As far as is known, this study is the first to provide evidence for such an association.

Forkhead box H1 (FOXH1) sequence variants in ventricular septal defect.

OBJECTIVE: To investigate the possible genetic role of FOXH1 in the pathogenesis of ventricular septal defect (VSD). PATIENTS AND METHODS: 301 VSD Chinese patients and 111 Chinese patients with the other subtypes of congenital heart defects were investigated for mutations in the FOXH1 gene by direct sequencing. RESULT: Four variants were found among the isolated VSD patients, including one pathogenic mutation (c.659_660ins.C).

Genetic variation of SAL-Like 4 (SALL4) in ventricular septal defect.

BACKGROUND: Ventricular septal defect (VSD) accounts for about half of congenital heart disease (CHD). SAL-Like 4 (SALL4) gene mutations have been identified to be the cause of Okihiro syndrome which is characterized by association limb and multiple other organ developmental defects including heart defect. METHODS: We screened SALL4 gene coding regions for mutations using DNA sequencing approach in 300 nonsyndromic VSD patients and 250 with no reported cardiac phenotype controls. RESULTS: We discovered two novel variants: c.586C>T (p.Arg196Trp) and c.2389A>T (p.Ser797Cys) in 300 nonsyndromic VSD patients. CONCLUSIONS: The two non-synonymous variations were located in the SALL4 evolutionarily conserved residues and the amino acid change may affect the function of SALL4. Our finding is the first to suggest that SALL4 may be a potential candidate gene of ventricular septal defect (VSD).

Lack of an association between CYP1A1 gene Ile462Val polymorphism and polycystic ovary syndrome in Chinese.

Polycystic ovary syndrome (PCOS) is a complex multi-factorial disorder involving a number of genetic and environmental factors. CYP1A1 (Cytochrome P450, family 1, subfamily A, polypeptide 1) gene, which belongs to Cytochrome P450 (CYP) super family, encodes a phase I cytochrome P450 enzyme, involved in the oxidative metabolism of estrogens. A recent study suggested that a common polymorphism Ile462Val of the CYP1A1 gene might be associated with PCOS development in Turkish women. To investigate a possible association between the CYP1A1 Ile462Val polymorphism and PCOS in Chinese women, we examined 205 PCOS patients and 177 healthy controls. All subjects were genotyped for CYP1A1. There was no statistical difference in CYP1A1 genotype and allele frequencies between PCOS cases and controls (chi(2) = 0.956, df = 2, P = 0.089 by genotype; chi(2) = 0.005, df = 1, P = 0.941 by allele). Compared with controls, there were no statistical difference in Val/Val genotype and Val allele frequency in the PCOS cases (4.9% vs. 5.1% by genotype; 51.7% vs. 52.0% by allele) (chi(2) = 0.009, df = 1, P = 0.926 by genotype; chi(2) = 0.005, df = 1, P = 0.941 by allele). Moreover, no association between CYP1A1 Ile462Val genotypes and metabolic parameters was observed in PCOS women. Our findings clearly indicated that this polymorphism does not represent an additional genetic risk factor for PCOS in Chinese women.