A Preterm Infant with Feeding Aspiration Diagnosed with BOR Syndrome, Confirmed Case by Whole-Genome Sequencing and Structural Variant Calling

Branchiootorenal (BOR) syndrome is a rare autosomal dominant inherited disease with a prevalence of approximately 1 in 40,000 newborns. This disease is characterized by hearing loss, preauricular pits, branchial fistulas or cysts, and renal dysplasia. We discovered a case of BOR syndrome in a premature 2-week-old female infant with a gestational age of 32 weeks and two days. She and her family had major symptoms and a family history of BOR. BOR syndrome was confirmed by whole-genome sequencing and structural variant calling, which revealed an EYA1 exon 5–6 deletion. The infant had recurrent sleep and feeding cyanosis with second branchial anomalies. Via videofluoroscopic swallowing study and a modified barium swallow test, penetration into the vocal cords was observed before and during swallowing when bottle feeding. This is the first report of a preterm infant early diagnosed with BOR syndrome in which deletion margin was accurately identified by whole-genome sequencing and structural variant calling in Republic of Korea

Rapid activation of CLOCK by Ca(2+)-dependent protein kinase C mediates resetting of the mammalian circadian clock

In mammals, immediate-early transcription of the Period 1 (Per1) gene is crucial for resetting the mammalian circadian clock. Here, we show that CLOCK is a real signalling molecule that mediates the serum-evoked rapid induction of Per1 in fibroblasts through the Ca(2+)-dependent protein kinase C (PKC) pathway. Stimulation with serum rapidly induced nuclear translocation, heterodimerization and Ser/Thr phosphorylation of CLOCK just before the surge of Per1 transcription. Serum-induced CLOCK phosphorylation was abolished by treatment with PKC inhibitors but not by other kinase inhibitors. Consistently, the interaction between CLOCK and PKC was markedly increased shortly after serum shock, and the Ca(2+)-dependent PKC isoforms PKCα and PKCγ phosphorylated CLOCK in vitro. Furthermore, phorbol myristic acetate treatment triggered immediate-early transcription of Per1 and also CLOCK phosphorylation, which were blocked by a Ca(2+)-dependent PKC inhibitor. These findings indicate that CLOCK activation through the Ca(2+)-dependent PKC pathway might have a substantial role in phase resetting of the circadian clock

Meal Time Shift Disturbs Circadian Rhythmicity along with Metabolic and Behavioral Alterations in Mice

<div><p>In modern society, growing numbers of people are engaged in various forms of shift works or trans-meridian travels. Such circadian misalignment is known to disturb endogenous diurnal rhythms, which may lead to harmful physiological consequences including metabolic syndrome, obesity, cancer, cardiovascular disorders, and gastric disorders as well as other physical and mental disorders. However, the precise mechanism(s) underlying these changes are yet unclear. The present work, therefore examined the effects of 6 h advance or delay of usual meal time on diurnal rhythmicities in home cage activity (HCA), body temperature (BT), blood metabolic markers, glucose homeostasis, and expression of genes that are involved in cholesterol homeostasis by feeding young adult male mice in a time-restrictive manner. Delay of meal time caused locomotive hyperactivity in a significant portion (42%) of subjects, while 6 h advance caused a torpor-like symptom during the late scotophase. Accordingly, daily rhythms of blood glucose and triglyceride were differentially affected by time-restrictive feeding regimen with concurrent metabolic alterations. Along with these physiological changes, time-restrictive feeding also influenced the circadian expression patterns of low density lipoprotein receptor (LDLR) as well as most LDLR regulatory factors. Strikingly, chronic advance of meal time induced insulin resistance, while chronic delay significantly elevated blood glucose levels. Taken together, our findings indicate that persistent shifts in usual meal time impact the diurnal rhythms of carbohydrate and lipid metabolisms in addition to HCA and BT, thereby posing critical implications for the health and diseases of shift workers.</p> </div

ANOVA <i>F</i> and <i>p</i> values for each physiological index.

<p>Significant differences (*<i>p</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 and ****<i>p</i><0.0001) are indicated in bold type.</p><p>ANOVA, analysis of variance; ns, not significant.</p

Daily rhythms of blood glucose and some metabolic parameters related to cholesterol homeostasis in mice fed time-restrictively.

<p>Young adult male C57BL/6J mice were first entrained to a 12∶12 LD photoperiodic cycle for two weeks. Then mice were fed time-restrictively for seven consecutive days as denoted in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044053#pone-0044053-g001" target="_blank">Figure 1</a>. Mice were sacrificed by cervical dislocation at the indicated ZT and whole blood samples were collected. Total cholesterol (A), HDL cholesterol (B), plasma triglyceride (C), blood glucose (D) levels were determined by specific kits obtained from Callegari^TM. All data are expressed as mean ± S.E.M. (n = 4–8). Statistical analyses are summarized in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044053#pone-0044053-t002" target="_blank">Table 2</a>.</p

ANOVA <i>F</i> and <i>p</i> values for BT and HCA rhythms.

<p>Significant differences (****<i>p</i><0.0001) are indicated in bold type.</p><p>ANOVA, analysis of variance.</p

Daily and circadian expressions of LDLR and some LDLR regulatory factors in the mouse liver.

<p>To determine daily expression patterns, young adult C57BL/6J male mice were entrained to a 12L:12D cycle for two weeks and liver samples were quickly obtained at the indicated ZT. For circadian sampling, mice were entrained to a 12L:12D cycle for two weeks and released to constant darkness (DD). On the second day after light-off, liver samples were obtained at the indicated circadian time (CT). RNA isolation, reverse transcription, and real-time polymerase chain reaction to measure specific messages for mouse <i>ldlr</i>, and LDLR regulatory factors. All mRNA levels were normalized to <i>tbp</i> mRNA levels. Data are expressed as mean ± S.E.M. (n = 8).</p

Experimental scheme.

<p>Young adult C57BL/6J male mice (8 weeks old) were first entrained to a 12∶12 LD photoperiodic cycle for two weeks with food and water available <i>ad libitum</i>. Then mice were randomly divided into three groups: food available <i>ad libitum</i> (AF), food available during the late day (ZT 6 to 11) (DF), and food available during the late night (ZT 18 to 23) (NF) with water available all the time. This time-restrictive feeding regimen was maintained either up to the end of each experiment or for 4 weeks and then returned to <i>ad libitum</i> feeding (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044053#pone-0044053-g002" target="_blank">Figure 2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044053#pone-0044053-g003" target="_blank">3</a>). Gray bar indicates major meal time of normal adult mouse (Figure S1A).</p

Effects of one week of time-restrictive feeding on the phases of Per1, LDLR, and LDLR regulatory factors gene expression in the mouse liver.

<p>Young adult male mice, entrained to a 12∶12 photoperiodic cycle, were fed time-restrictively for seven consecutive days as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044053#pone-0044053-g001" target="_blank">Figure 1</a>. On the 8^th day, mice were sacrificed at the indicated zeitgeber time (ZT) and liver samples were obtained. RNA isolation, reverse transcription, and real-time polymerase chain reaction were performed to measure specific messages for mouse <i>Per1</i>, <i>ldlr</i>, and LDLR regulatory factors. All mRNA levels were normalized to <i>tbp</i> mRNA levels. Data are expressed as mean ± S.E.M. (n = 4).</p