Sibling Controls Research Articles

Ankle2 deficiency-associated microcephaly and spermatogenesis defects in zebrafish are alleviated by heterozygous deletion of vrk1

Autosomal recessive primary microcephaly (MCPH) is a rare congenital disorder characterized by a below average brain volume at birth and is associated with neurodevelopmental disorders such as growth retardation and intellectual disability. Mutations in ANKLE2 have been identified as one of the causes of MCPH (MCPH16). ANKLE2 is a target molecule of the Zika virus NS4a protein that interferes with ANKLE2 function, resulting in severe microcephaly. ANKLE2 is essential for organizing the nuclear envelope and chromatin structures during the mitotic-end process via barrier to autointegration factor (BAF) dephosphorylation. However, the precise mechanism by which the loss of ANKLE2 function causes the pathogenesis of microcephaly remains unclear. In this study, we generated Ankle2-deficient zebrafish (ankle2−/−) with a significant reduction in brain size compared with that of their control siblings. The ankle2−/− brain showed a significant decrease in the number of radial glial progenitor cells, suggesting that Ankle2 deficiency in zebrafish causes neurogenesis defects. Furthermore, ankle2−/− male zebrafish showed infertility owing to defects in spermatogenesis. Notably, microcephaly was overcome by vrk1 morpholino knockdown or vrk1 heterozygous deletion. In addition, spermatogenesis in ankle2−/− zebrafish males was partially restored by the vrk1 heterozygous deletion, although infertility was not resolved. These results indicate that ANKLE2 and VRK1 coordinate with each other for BAF phosphorylation to maintain normal mitosis during neurogenesis and spermatogenesis.

Kidney disease in very long-term survivors of Wilms tumor: A nationwide cohort study with sibling controls.

Survival after Wilms tumor has significantly increased and focus on late effects has become increasingly important. However, knowledge about long-term renal function in survivors of Wilms tumor is missing. Our aim was to investigate evidence of kidney disease in 20- or more-year survivors of Wilms tumor in a clinical setting, with siblings as comparisons. In this cross-sectional study, we established a cohort of Danish 20-plus-year survivors of Wilms tumor and siblings as controls. Participants answered a comprehensive health questionnaire supplemented by measurements of estimated glomerular filtration rate (eGFR), urine albumin-to-creatinine ratio, and blood pressure and were categorized according to the chronic kidney disease classification. Multiple linear regression analysis, taking family membership into account, was used to describe the differences in eGFR. Logistic regression analysis was performed to describe risk factors for the development of kidney disease. We included 99 survivors of Wilms tumor and 38 sibling controls with a median of 37 years of follow-up. The eGFR of Wilms tumor survivors was 13ml/min/1.73 m2 (95% CI -20; -5) lower when compared to sibling control. Evidence of kidney disease, with risk factors as hypertension and diabetes, was found in 19% of the Wilms tumor survivors and 2% developed end-stage renal disease. Ninety-two percent of the Wilms tumor survivors had an eGFR >60ml/min/1.732 . Long-term Wilms tumor survivors have on average a significantly decreased renal function along with the increased prevalence of kidney disease and end-stage renal disease when compared to sibling controls. Still, most survivors had kidney function within the normal range.

Association of mitochondrial DNA content, heteroplasmies and inter-generational transmission with autism

Mitochondria are essential for brain development. While previous studies linked dysfunctional mitochondria with autism spectrum disorder (ASD), the role of the mitochondrial genome (mtDNA) in ASD risk is largely unexplored. This study investigates the association of mtDNA heteroplasmies (co-existence of mutated and unmutated mtDNA) and content with ASD, as well as its inter-generational transmission and sex differences among two independent samples: a family-based study (n = 1,938 families with parents, probands and sibling controls) and a prospective birth cohort (n = 997 mother-child pairs). In both samples, predicted pathogenic (PP) heteroplasmies in children are associated with ASD risk (Meta-OR = 1.56, P = 0.00068). Inter-generational transmission of mtDNA reveals attenuated effects of purifying selection on maternal heteroplasmies in children with ASD relative to controls, particularly among males. Among children with ASD and PP heteroplasmies, increased mtDNA content shows benefits for cognition, communication, and behaviors (P ≤ 0.02). These results underscore the value of exploring maternal and newborn mtDNA in ASD.

Loss of starch synthase IIIa changes starch molecular structure and granule morphology in grains of hexaploid bread wheat

Starch synthase III plays a key role in starch biosynthesis and is highly expressed in developing wheat grains. To understand the contribution of SSIII to starch and grain properties, we developed wheat ssIIIa mutants in the elite cultivar Cadenza using in silico TILLING in a mutagenized population. SSIIIa protein was undetectable by immunoblot analysis in triple ssIIIa mutants carrying mutations in each homoeologous copy of ssIIIa (A, B and D). Loss of SSIIIa in triple mutants led to significant changes in starch phenotype including smaller A-type granules and altered granule morphology. Starch chain-length distributions of double and triple mutants indicated greater levels of amylose than sibling controls (33.8% of starch in triple mutants, and 29.3% in double mutants vs. 25.5% in sibling controls) and fewer long amylopectin chains. Wholemeal flour of triple mutants had more resistant starch (6.0% vs. 2.9% in sibling controls) and greater levels of non-starch polysaccharides; the grains appeared shrunken and weighed ~ 11% less than the sibling control which was partially explained by loss in starch content. Interestingly, our study revealed gene dosage effects which could be useful for fine-tuning starch properties in wheat breeding applications while minimizing impact on grain weight and quality.

PB1751: ALTERATIONS IN THE GUT MICROBIOME IN PEDIATRIC, ADOLESCENT, AND YOUNG ADULT PATIENTS WITH ACUTE LEUKEMIA

Background: Gut microbiome is involved in the progression of various diseases and may affect the human immune system. The composition of gut microbiome and its change upon anti-leukemic therapy in pediatric, adolescent, and young adult patients with acute leukemia is poorly understood. Aims: We performed a longitudinal analysis of gut microbiome composition to assess associations between gut microbial dynamics and clinical outcomes in pediatric, adolescent, and young adult patients with acute leukemia during chemotherapy and/or hematopoietic cell transplantation (HCT). And also, we aimed to compare the gut microbiome composition of leukemia patients with their healthy sibling controls. Methods: The bacterial 16S rRNA gene was analyzed by high-depth sequencing to determine the diversity and composition of the microbiome. Data collected between August 2020 and September 2021. The median age of patients was 17.1 years (range from 3.5 years to 34.1 years). Seventy-nine stool samples from 15 patients (10 acute lymphoblastic leukemia and 5 acute myeloid leukemia) and 9 stool samples from 9 siblings were analyzed in this study. Relative abundance, overall richness, various diversity indexes were calculated. Results: At diagnosis, microbial diversity index of the patient group was lower than that of their siblings. Clostridia and Lachnospiraceae were reduced in patients, but Actinobacteria were more abundant in patients compared to sibling controls. The Firmicutes/Bacteroidetes (F/B) ratio was not different between patients and their siblings. In patients, microbial diversity decreased after induction chemotherapy, but it recovered to a level similar to that of sibling controls after consolidation chemotherapy. The F/B ratio significantly increased after induction, but it decreased to a similar value at diagnosis after consolidation. Microbial diversity index at diagnosis was significantly lower in patients who experienced fever and/or infection during induction compared to patients who did not. A baseline abundance of Proteobacteria predicted fever and/or infection during induction chemotherapy. In patients underwent HCT, compared to the values before HCT, the number of identified species has decreased to about half at day 28 after HCT and decreased further at day 56 after HCT. However, it recovered to pre-HCT level at day 90 after HCT. Diversity index significantly decreased at day 0 of HCT compared to that of pre-HCT, but it recovered to a level similar to that of pre-HCT at day 90 after HCT. At day 0 of HCT, patients who experienced fever and/or acute graft versus host disease (aGVHD) within day 100 of HCT showed abundant Bacteroidetes with median F/B ratio of 0.1, while patients who did not showed low level of Bacteriodetes with median F/B ratio of 1.4. Diversity index at day 0 of HCT was not different between patients who experienced fever and/or aGVHD and patients who did not. Summary/Conclusion: In pediatric, adolescent, and young adult patients undergoing therapy for acute leukemia, decreased microbial diversity and abundance of Proteobacteria at diagnosis were associated with fever and/or infection during induction chemotherapy. Abundance of Bacteroidetes at day 0 of HCT predicted subsequent fever and/or acute GVHD. Better understanding of the gut microbiome could provide actionable information in guiding precision medicine.

Prematurity Is a Chronic Disease

Few studies have investigated neuroradiographic findings of preterm birth from a clinical perspective into adulthood, or used a sibling comparison to demonstrate these differences. In this volume of The Journal, Kuula et al report deployment of MRI to assess brain abnormalities and volumetric differences in a cohort of adults born preterm at very low birth weight (VLBW), using siblings as controls. At mean age 29 years, the investigators found significantly more structural abnormalities in VLBW subjects than control siblings, 37% vs. 13%, with periventricular leukomalacia the most common finding. The VLBW group had smaller absolute brain volumes and, after adjusting for estimated intracranial volume, also larger ventricles, less gray matter, and smaller thalami, caudate nuclei, right hippocampus, and left pallidum. No differences in total white matter were detected. While other researchers have documented chronic white matter abnormalities in adults following prematurity, the chronic gray matter changes here are intriguing and require further study. Perhaps more notable is the persistence of sequelae far into adult life. Children with cerebral palsy and visual impairments were excluded from this study, but other neurodevelopmental differences in this population may be associated with these abnormalities. Children born premature are already known to be at increased risk of hypertension and chronic cardiovascular disease, chronic pulmonary disease, diabetes mellitus, and even premature death. The findings of Kuula et al underscore that prematurity is a chronic disease also affecting the central nervous system. There are no guidelines for the long-term medical follow-up of people born preterm. Meanwhile there are survivorship guidelines for prevalent yet less common conditions of childhood, such as cancer or liver transplantation. With about 10% of US births categorized as premature and over 1% VLBW, perhaps it is time give more attention to the investigation and management of chronic diseases in adulthood that arise from prematurity. Article page 48

Premature Neural Progenitor Cell Differentiation Into Astrocytes in Retinoic Acid-Induced Spina Bifida Rat Model.

During embryonic spinal cord development, neural progenitor cells (NPCs) generate three major cell lines: neurons, oligodendrocytes, and astrocytes at precise times and locations within the spinal cord. Recent studies demonstrate early astrogenesis in animal models of spina bifida, which may play a role in neuronal dysfunction associated with this condition. However, to date, the pathophysiological mechanisms related to this early astrocytic response in spina bifida are poorly understood. This study aimed to characterize the development of early astrogliosis over time from Pax6+, Olig2+, or Nkx2.2+ NPCs using a retinoic acid-induced spina bifida rat model. At three gestational ages (E15, E17, and E20), spinal cords from fetuses with retinoic acid-induced spina bifida, their healthy sibling controls, or fetuses treated with the vehicle control were analyzed. Results indicated that premature astrogliosis and astrocytic activation were associated with an altered presence of Pax6+, Olig2+, and Nkx2.2+ NPCs in the lesion compared to the controls. Finally, this response correlated with an elevation in genes involved in the Notch-BMP signaling pathway. Taken together, changes in NPC patterning factor expression with Notch-BMP signaling upregulation may be responsible for the altered astrogenesis patterns observed in the spinal cord in a retinoic acid-induced spina bifida model.

The Investigation of Cognitive Functions and Clinical High Risk Status for Psychosis in First-Degree Relatives of Patients with Substance Induced Psychotic Disorder

IntroductionThe etiology of substance-induced psychotic disorder (SIPD) is an important research area to study.ObjectivesIt is aimed to investigate clinical risk status for psychosis, schizotypal features and neurocognitive functions in siblings of the patients who have been diagnosed as SIPD and who have no family history of psychotic spectrum disorder.MethodsThis study included 41 healthy siblings of patients who have been diagnosed as SIPD according to DSM-V and 41 healthy controls without family history of psychiatric disorders (matched on age, gender, and years of education). The data collected with sociodemographic and clinical data form, Digid Span Test, Trail Making Test A, Trail Making Test B, Verbal Fluency Test and Stroop Test, Comprehensive Assesment of At-Risk Mental States (CAARMS) and Structured Interview for Schizotypy-Revised.ResultsIt is determined that %41.5 of siblings and %7.3 of healthy controls are in one of the clinical high risk groups for psychosis according to CAARMS. There is significant difference in Trail Making Test A error and Trail Making Test B error and correction, verbal fluency test- lexical fluency-perseveration mean scores between siblings of patients and healthy controls.ConclusionsSiblings of patients with SIPD have more schizotypal features than healthy control group and they take part more frequent in one of high risk group for psychosis. Schizotypal features are known as trait factor and show genetic predisposition. Siblings who are in high risk groups have more schizotypal features and it may point that predisposition to psychosis is more related to underlying genetic predisposition than environmental factors and social stressors.DisclosureNo significant relationships.

Job loss and fetal growth restriction: identification of critical trimesters of exposure

Previous research suggests that job loss in a household during pregnancy may perturb fetal growth. However, this work often cannot rule out unmeasured confounding due to selection into job loss. Recent work using data on exogenous job loss (due to a plant closure) finds that a father's unexpected job loss during his spouse's pregnancy increases the risk of a low weight birth. Using a unique set of linked registries in Denmark, we build on this work and examine whether associations between a father's unexpected job loss and low birthweight differ by trimester of in utero exposure. We additionally examine trimester-specific associations of job loss with small-for-gestational-age, a proxy for restricted fetal growth, which may cause low birthweight. We apply a sibling control design to over 1.4 million live births in Denmark, 1980 to 2017, to examine whether this plausibly exogenous form of job loss corresponds with increased risk of low weight or small-for-gestational-age births, depending on the timing of displacement in the first, second, or third trimester. Results indicate an elevated risk of low birthweight (OR=1.80, 95% CI: 1.24, 2.62) and small-for-gestational-age (OR=1.40, 95% CI: 1.02, 1.93) among gestations exposed to job loss in the second trimester of pregnancy. Sensitivity analyses using continuous outcome measures (e.g., birthweight in grams, birthweight for gestational age percentile) and maternal fixed effects analyses produce substantively similar inference. Findings support the notion that unexpected job loss may affect fetal growth and that the second trimester in particular appears sensitive to this external stressor.

The Role of Glutamatergic Signaling in Central Noradrenergic Neurons in Respiratory Control

Central noradrenergic (NA) neurons are key constituents of the respiratory homeostatic network. NA dysfunction is implicated in several developmental respiratory disorders such as the fatal Sudden Infant Death Syndrome (SIDS) and Rett Syndrome, in which abnormal chemosensory function is thought to play a role. However, which NA populations modulate respiratory chemoreflexes and the underlying mechanisms are still not fully understood. Histological evidence suggests that glutamate is co‐transmitted in subsets of brainstem NA neurons, some of which may contribute to regulating chemoreflexes. However, the extent of glutamate co‐transmission in the central NA system remains uncharacterized. Further, it remains unclear what role NA‐based glutamatergic signaling plays in various aspects of respiratory homeostasis.To anatomically map potential glutamate‐expressing NA neurons, we characterized the expression profile of all three known of Vesicular Glutamate Transporters (Vglut1, Vglut2 and Vglut3) in NA neurons by intersectional genetic fate mapping. For each Vglut1/2/3 transporter, acorresponding Cre knock‐in recombinase line was combined with DBH‐p2a_Flpomice. These Vglut1/2/3_Cre; DBH‐p2a_Flpocompound lines were crossed with the RC::F_TdTomato_P_GFPintersectional reporter mice. In each of the three intersectional reporter crosses, NA neurons co‐expressing either Vglut1, Vglut2, or Vglut3 are labeled by green fluorescent protein (GFP) while NA neurons without any Vglut1/2/3 expression are labelled by red fluorescent protein (tdTomato). Our preliminary fate maps, which reflect the cumulative expression profiles of glutamate markers in NA neurons throughout the development and lifetime of the mouse, found that about 80% NA neurons express Vglut2 at some point during their development and posterior medullary NA neurons express Vglut3 as well.Prior studies suggest that Vglut2‐based glutamatergic signaling may play a role in respiratory control. To determine what physiological role, if any, Vglut2‐based glutamatergic signaling may play in adult respiratory homeostasis, we conditionally ablated Vglut2 in central NA neurons by crossing DBH‐Cre/+with Vglut2flox/flox mice. Mutants and sibling controls (6‐8 weeks) were assessed for respiratory function by whole‐body barometric plethysmography. Ventilation and VO2 were assessed under room air (21%O2,79%N2), hypercapnia (21%O2, 74/72/69%N2, 5/7/10%CO2) and hypoxia (10% O2, 90% N2). Our preliminary data indicates that loss of Vglut2 in NA neurons does not significantly impact breathing under room air, hypercapnia, or hypoxia. Thus, our data suggests that Vglut2‐based glutamatergic signaling within the central NA system is dispensable for normal baseline breathing and hypercapnic, and hypoxic chemosensory reflexes.

Visual processing difficulties in children with NF1. A common but not widely recognized underlying cause of reading difficulties

Neurofibromatosis type 1 (NF1) is one of the most frequent multi-systemic autosomal dominant rare genetic disorder with an incidence as high as 1/2000 and prevalence of 1/3000 births. NF1 is caused by pathogenic variants of the NF1 gene located on chromosome 17q11.2 [ [1] Ottenhoff M.J. Rietman A.B. Mous S.E. Plasschaert E. Gawehns D. Brems H. Oostenbrink R. Van Minkelen R. Nellist M. Schorry E. Legius E. Moll H.A. Elgersma Y. Examination of the genetic factors underlying the cognitive variability associated with neurofibromatosis type 1. Genet. Med. 2020; 22: 889-897https://doi.org/10.1038/s41436-020-0752-2 Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar ]. Of the affected individuals, 50% have a sporadic disease which causes challenges for early diagnosis. Up to 80% of children aged 6–18 years with NF1 present with moderate to severe impairment in one or more areas of cognitive functioning, and they are among the most important NF1 manifestations during school age [ [2] Johansson E. Kallionpää Böckerman P. Peltonen J. Peltonen S. A rare disease and education: neurofibriomatosis type 1 decreases educational attainment. Clin. Genet. 2021; 99: 529-539 Crossref PubMed Scopus (6) Google Scholar , [3] Van der Vaart T. Plasschaert E. Rietman A.B. Renard M. Oostenbrink R. Vogels A. de Wit M.C.Y. Descheemaeker M. Vergouwe Y. Catsman-Berrevoets C.E. Legius E. Elgersma Y. Moll H.A. Simvastatin for cognitive deficits and behavioural problems in patients with neurofibromatosis type 1 (NF1-SIMCODA): a randomised, placebo-controlled trial. Lancet Neurol. 2013; 12: 1076-1083 Abstract Full Text Full Text PDF PubMed Scopus (90) Google Scholar ]. The average intelligence quotient (IQ) is 10–15 points lower in these children than in population or sibling control groups. Learning disabilities in reading, writing and mathematics have been reported in about 50% of children [ [2] Johansson E. Kallionpää Böckerman P. Peltonen J. Peltonen S. A rare disease and education: neurofibriomatosis type 1 decreases educational attainment. Clin. Genet. 2021; 99: 529-539 Crossref PubMed Scopus (6) Google Scholar ]. Furthermore, achievements may be undermined by attention-deficit–hyperactivity disorder (ADHD), difficulties in social functioning associated with autism spectrum disorder (ASD) and motor difficulties [ 2 Johansson E. Kallionpää Böckerman P. Peltonen J. Peltonen S. A rare disease and education: neurofibriomatosis type 1 decreases educational attainment. Clin. Genet. 2021; 99: 529-539 Crossref PubMed Scopus (6) Google Scholar , 3 Van der Vaart T. Plasschaert E. Rietman A.B. Renard M. Oostenbrink R. Vogels A. de Wit M.C.Y. Descheemaeker M. Vergouwe Y. Catsman-Berrevoets C.E. Legius E. Elgersma Y. Moll H.A. Simvastatin for cognitive deficits and behavioural problems in patients with neurofibromatosis type 1 (NF1-SIMCODA): a randomised, placebo-controlled trial. Lancet Neurol. 2013; 12: 1076-1083 Abstract Full Text Full Text PDF PubMed Scopus (90) Google Scholar , 4 Krab L.C. Aarsen F.K. de Goede-Bolder A. Catsman-Berrevoets C.E. Arts W.F.A. Moll H.A. Elgersma Y. Impact of neurofibromatosis type 1 on school performance. J. Child Neurol. 2008; 23: 1002-1010 Crossref PubMed Scopus (93) Google Scholar ]. Cognitive and behavioural deficits lead to lower academic achievement and loss of quality of life, persisting into adulthood [ [2] Johansson E. Kallionpää Böckerman P. Peltonen J. Peltonen S. A rare disease and education: neurofibriomatosis type 1 decreases educational attainment. Clin. Genet. 2021; 99: 529-539 Crossref PubMed Scopus (6) Google Scholar ]. 40% of children with NF1 need special education and only approximately 10% of children with NF1 do not have any problems with school performance and also do not need remedial teaching [ [4] Krab L.C. Aarsen F.K. de Goede-Bolder A. Catsman-Berrevoets C.E. Arts W.F.A. Moll H.A. Elgersma Y. Impact of neurofibromatosis type 1 on school performance. J. Child Neurol. 2008; 23: 1002-1010 Crossref PubMed Scopus (93) Google Scholar ].

IgA deficiency destabilizes immunological homeostasis towards intestinal microbiota and increases the risk of systemic immune dysregulation

Abstract Mammals produce large quantities of mucosal and systemic antibodies that maintain the intestinal barrier, shape the intestinal microbiome and promote lifelong mutualism with commensal microbes. Here, we developed an integrated host-commensal approach combining microbial flow cytometry and 16S rRNA gene sequencing to define the population of microbes that induce mucosal and systemic antibodies with CyTOF analysis in pediatric selective Immunoglobulin A (IgA) deficient and household control siblings to determine the impacts of IgA deficiency on host cellular immune phenotype. In healthy controls, mucosal secretory IgA and IgM antibodies coat an overlapping subset of microbes, predominantly Firmicutes and Proteobacteria. Serum IgG antibodies target a similar consortium of fecal microbes, revealing connections between mucosal and systemic antibody networks. Furthermore, we find broad systemic immune dysregulation in a subset of children and mice lacking IgA, including enhanced IgG targeting of fecal microbiota, elevated levels of inflammatory and allergic cytokines and alterations in T cell activation state. Thus, IgA tunes systemic interactions between the host and commensal microbiota. Understanding how IgA affects baseline immune tone has implications for predicting and preventing autoimmune, inflammatory and allergic diseases, as well as providing improved prognostic guidance to patients with IgA deficiency. Supported by CHOP Microbiome Pilot Grant K08AI135091 Burroughs Wellcome Fund, the American Academy of Allergy, Asthma, and Immunology, the Immune Deficiency Foundation, the Primary Immune Deficiency Treatment Consortium and Chan Zuckerberg Initiative

Clinical characterization of familial hypercholesterolemia due to an amish founder mutation in Apolipoprotein B

BackgroundFamilial hypercholesterolemia (FH) due to a founder variant in Apolipoprotein B (ApoBR3500Q) is reported in 12% of the Pennsylvania Amish community. By studying a cohort of ApoBR3500Q heterozygotes and homozygotes, we aimed to characterize the biochemical and cardiac imaging features in children and young adults with a common genetic background and similar lifestyle.MethodsWe employed advanced lipid profile testing, carotid intima media thickness (CIMT), pulse wave velocity (PWV), and peripheral artery tonometry (PAT) to assess atherosclerosis in a cohort of Amish ApoBR3500Q heterozygotes (n = 13), homozygotes (n = 3), and their unaffected, age-matched siblings (n = 9). ApoBR3500Q homozygotes were not included in statistical comparisons.ResultsLDL cholesterol (LDL-C) was significantly elevated among ApoBR3500Q heterozygotes compared to sibling controls, though several ApoBR3500Q heterozygotes had LDL-C levels in the normal range. LDL particles (LDL-P), small, dense LDL particles, and ApoB were also significantly elevated among subjects with ApoBR3500Q. Despite these differences in serum lipids and particles, CIMT and PWV were not significantly different between ApoBR3500Q heterozygotes and controls in age-adjusted analysis.ConclusionsWe provide a detailed description of the serum lipids, atherosclerotic plaque burden, vascular stiffness, and endothelial function among children and young adults with FH due to heterozygous ApoBR3500Q. Fasting LDL-C was lower than what is seen with other forms of FH, and even normal in several ApoBR3500Q heterozygotes, emphasizing the importance of cascade genetic testing among related individuals for diagnosis. We found increased number of LDL particles among ApoBR3500Q heterozygotes but an absence of detectable atherosclerosis.

Inhibition of miR-128 Enhances Vocal Sequence Organization in Juvenile Songbirds.

The molecular mechanisms underlying learned vocal communication are not well characterized. This is a major barrier for developing treatments for conditions affecting social communication, such as autism spectrum disorder (ASD). Our group previously generated an activity-dependent gene expression network in the striatopallidal song control nucleus, Area X, in adult zebra finches to identify master regulators of learned vocal behavior. This dataset revealed that the two host genes for microRNA-128, ARPP21 and R3HDM1, are among the top genes whose expression correlates to how much birds sing. Here we examined whether miR-128 itself is behaviorally regulated in Area X and found that its levels decline with singing. We hypothesized that reducing miR-128 during the critical period for vocal plasticity would enhance vocal learning. To test this, we bilaterally injected an antisense miR-128 construct (AS miR-128) or a control scrambled sequence into Area X at post-hatch day 30 (30 d) using sibling-matched experimental and control pupils. The juveniles were then returned to their home cage and raised with their tutors. Strikingly, inhibition of miR-128 in young birds enhanced the organization of learned vocal sequences. Tutor and pupil stereotypy scores were positively correlated, though the correlation was stronger between tutors and control pupils compared to tutors and AS miR-128 pupils. This difference was driven by AS miR-128 pupils achieving higher stereotypy scores despite their tutors’ lower syntax scores. AS miR-128 birds with tutors on the higher end of the stereotypy spectrum were more likely to produce songs with faster tempos relative to sibling controls. Our results suggest that low levels of miR-128 facilitate vocal sequence stereotypy. By analogy, reducing miR-128 could enhance the capacity to learn to speak in patients with non-verbal ASD. To our knowledge, this study is the first to directly link miR-128 to learned vocal communication and provides support for miR-128 as a potential therapeutic target for ASD.

Characterizing Emotion Recognition and Theory of Mind Performance Profiles in Unaffected Siblings of Autistic Children.

Emotion recognition skills and the ability to understand the mental states of others are crucial for normal social functioning. Conversely, delays and impairments in these processes can have a profound impact on capability to engage in, maintain, and effectively regulate social interactions. Therefore, this study aimed to compare the performance of 42 autistic children (Mage = 8.25 years, SD = 2.22), 45 unaffected siblings (Mage = 8.65 years, SD = 2.40), and 41 typically developing (TD) controls (Mage = 8.56 years, SD = 2.35) on the Affect Recognition (AR) and Theory of Mind (TOM) subtests of the Developmental Neuropsychological Assessment Battery. There were no significant differences between siblings and TD controls. Autistic children showed significantly poorer performance on AR when compared to TD controls and on TOM when compared to both TD controls and unaffected siblings. An additional comparison of ASD, unaffected sibling and TD control subsamples, matched on full-scale IQ, revealed no group differences for either AR or TOM. AR and TOM processes have received less research attention in siblings of autistic children and remain less well characterized. Therefore, despite limitations, findings reported here contribute to our growing understanding of AR and TOM abilities in siblings of autistic children and highlight important future research directions.

Oral Health Status of Children Living with Type 1 Diabetes Mellitus.

Background: Diabetes is a well-known predisposing factor for oral diseases, so prevention in an early age is mandatory. Objective: To provide oral screening for children living with type 1 diabetes. We aimed to investigate the oral and general health indexes of T1DM children and compare these data to healthy siblings and controls. Methods: In this cross-sectional study, 120 DM patients and 78 siblings, thereafter 80 DM children and 95 controls, took part. A detailed questionnaire, panoramic radiographs, and lateral cephalograms were obtained in every orthodontic consultation. We used Pearson’s chi-square test for statistical analysis and compared the data of the study and control groups. Results: The oral health values of DM children were significantly better (DMF-T 0.83–1.3) than the national average (3.8–4.5). A total of 75% (n = 60) of the children needed orthodontic treatment for orthodontic or skeletal anomalies. The prevalence of skeletal anomalies was significantly (p < 0.05) higher among patients with diabetes mellitus (DM) than in the control group. The frequency of coeliac disease was significantly elevated compared to any literature data (1–3.5%) in the study (15%) and in the control sibling group (13%). Conclusions: Co-morbidities such as CD should get more attention as a prognostic factor for a future higher incidence of diabetes. T1DM children can be motivated and health-conscious patients with excellent oral hygiene and dental status. Orthodontic treatment can help eliminate the oral complications of DM. Special diabetes ambulances may help provide oral care for patients with DM.

Characterization of wheat lacking B-type starch granules

The physicochemical and agronomic properties of a new form of bread wheat lacking B-type starch granules (BlessT) were assessed. Three BlessT mutant lines made by combining homoeologous deletions of BGC1, a gene responsible for the control of B-granule content, were compared with two sibling lines with normal starch phenotype and the parent line, cv. Paragon. Quantification of starch granule size and number in developing grain confirmed the lack of small, B-type starch granules throughout development in BlessT. Most starch, flour, grain and loaf characteristics did not vary between BlessT and the wild type sibling controls. However, BlessT starches had higher water absorption, reduced grain hardness and higher protein content, and dough made from BlessT flour required more water and had increased elasticity. Despite the lack of B-granules, BlessT lines do not display a significant decrease in total starch content suggesting that it should be possible to produce commercial wheat varieties that lack B-type starch granules without compromising yield. These findings support the potential utility of this novel type of wheat as a specialist crop in applications ranging from bread making and alcohol production to improved industrial starch products.

Impact of Risk-Stratified Therapy on Health Status in Survivors of Childhood Acute Lymphoblastic Leukemia: A Report from the Childhood Cancer Survivor Study.

Prior studies have identified that survivors of childhood acute lymphoblastic leukemia (ALL) report poor health status. It is unknown how risk-stratified therapy impacts the health status of ALL survivors. We estimated and compared the prevalence of self-reported poor health status among adult (≥18 years) survivors of childhood ALL diagnosed at age <21 years from 1970 to 1999 and sibling controls, excluding proxy reports. Therapy combinations defined treatment groups representative of 1970s therapy (70s), standard- and high-risk 1980s and 1990s therapy (80sSR, 80sHR, 90sSR, 90sHR), and relapse/bone marrow transplant (R/BMT). Log-binomial models, adjusted for clinical and demographic factors, compared outcomes between groups using prevalence ratios (PR) with 95% confidence intervals (CI). Among 5,119 survivors and 4,693 siblings, survivors were more likely to report poor health status in each domain including poor general health (13.5% vs. 7.4%; PR = 1.92; 95% CI, 1.69-2.19). Compared with 70s, 90sSR and 90sHR were less likely to report poor general health (90sSR: PR = 0.75; 95% CI, 0.57-0.98; 90sHR: PR = 0.58; 95% CI, 0.39-0.87), functional impairment (90sSR: PR = 0.56; 95% CI, 0.42-0.76; 90sHR: PR = 0.63; 95% CI, 0.42-0.95), and activity limitations (90sSR: 0.61; 95% CI, 0.45-0.83; 90sHR: PR = 0.59; 95% CI, 0.38-0.91). An added adjustment for chronic conditions in multivariable models partially attenuated 90sSR risk estimates. Risk-stratified ALL therapy has succeeded in reducing risk for poor general health, functional impairment, and activity limitations among more recent survivors of standard- and high-risk therapy. Future research into the relationship between risk-stratified therapy, health status, and late health outcomes may provide new opportunities to further improve late morbidity among survivors.

Leptin mutation and mycobacterial infection lead non-synergistically to a similar metabolic syndrome

IntroductionThe leptin signaling pathway plays an important role as a key regulator of glucose homeostasis, metabolism control and systemic inflammatory responses. However, the metabolic effects of leptin on infectious diseases, for example tuberculosis (TB), are still little known.ObjectivesIn this study, we aim to investigate the role of leptin on metabolism in the absence and presence of mycobacterial infection in zebrafish larvae and mice.MethodsMetabolites in entire zebrafish larvae and the blood of mice were studied using high-resolution magic-angle-spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy and mass spectrometry, respectively. For transcriptome studies of zebrafish larvae, deep RNA sequencing was used.ResultsThe results show that leptin mutation leads to a similar metabolic syndrome as caused by mycobacterial infection in the two species, characterized by the decrease of 11 amine metabolites. In both species, this metabolic syndrome was not aggravated further when the leptin mutant was infected by mycobacteria. Therefore, we conclude that leptin and mycobacterial infection are both impacting metabolism non-synergistically. In addition, we studied the transcriptomes of lepbibl54 mutant zebrafish larvae and wild type (WT) siblings after mycobacterial infection. These studies showed that mycobacteria induced a very distinct transcriptome signature in the lepbibl54 mutant zebrafish compared to WT sibling control larvae. Furthermore, lepbibl55 Tg (pck1:luc1) zebrafish line was constructed and confirmed this difference in transcriptional responses.ConclusionsLeptin mutation and TB lead non-synergistically to a similar metabolic syndrome. Moreover, different transcriptomic responses in the lepbibl54 mutant and TB can lead to the similar metabolic end states.

Elevated neurofibrillary tau levels emerge with subthreshold Aβ accumulation in Down syndrome

AbstractBackgroundAdults with Down syndrome (DS) are genetically predisposed to Alzheimer’s disease (AD) and accumulate beta‐amyloid plaques (Aβ) early in life. The aim of this study was to evaluate neurofibrillary tau deposition in DS between groups of Aβ negative (A‐) and subthreshold Aβ accumulators (subthreshold A+).MethodA total of 130 adults with DS (age=36.5 [6.77] years) and 40 healthy, non‐DS sibling controls (43.2 [12.6] years) underwent T1w‐MRI, [C‐11]PiB and [F‐18]AV‐1451 PET scans. MRI images were processed using FreeSurfer v5.3.0 to generate ROI masks encompassing the Braak staging of tau pathology. PiB and AV‐1451 SUVr images were generated using a cerebellar gray matter reference region. Global Aβ burden was calculated using the amyloid load metric (AβL). Of the 130 DS adults, N=26 were classified as subthreshold A+ (13.3 &lt; AβL &lt; 20), and the remaining 104 were classified as A‐ (AβL &lt; 13.3). AV‐1451 images were averaged for each group, and the difference was taken to generate a ΔSUVr image. Regional tau was assessed using AV‐1451 SUVr Z‐scores relative to the sibling control group. Partial volume correction was performed on AV‐1451 SUVrs in the Braak I‐II regions using the geometric transfer matrix method. Braak regional SUVr Z‐scores were then compared across the two groups using Student’s t‐tests while adjusting for imaging site. The analysis was then repeated using an age‐matched sample of A‐ (N=68) and subthreshold A+ (N=26) DS.ResultThe SUVr difference image revealed higher AV‐1451 uptake in Braak regions I‐III for the subthreshold A+ group (Figure 1). Student’s t‐tests revealed significantly higher AV‐1451 uptake in Braak regions I‐III for the subthreshold A+ group (Table 1). No difference in AV‐1451 uptake was observed between groups in Braak regions IV‐VI (Table 1). Higher AV‐1451 uptake in Braak regions I‐III was also observed in the subthreshold A+ group when compared to the age‐matched A‐ group (Table 2).ConclusionThese results show that subthreshold Aβ in DS is accompanied by elevated tau in the early Braak stage regions. These findings indicate that there is a short latency between the onset of Aβ and the spread of neurofibrillary tau in DS.