Neonatal Stage Research Articles

Prenatal smoke effect on mouse offspring Igf1 promoter methylation from fetal stage to adulthood is organ and sex specific.

Prenatal smoke exposure (PSE) is associated with reduced birth weight, impaired fetal development, and increased risk for diseases later in life. Changes in DNA methylation may be involved, as multiple large-scale epigenome-wide association studies showed that PSE is robustly associated with DNA methylation changes in blood among offspring in early life. Insulin-like growth factor-1 (IGF1) is important in growth, differentiation, and repair processes after injury. However, no studies investigated the organ-specific persistence of PSE-induced methylation change of Igf1 into adulthood. Based on our previous studies on the PSE effect on Igf1 promoter methylation in fetal and neonatal mouse offspring, we now have extended our studies to adulthood. Our data show that basal Igf1 promoter methylation generally increased in the lung but decreased in the liver (except for 2 persistent CpG sites in both organs) across three different developmental stages. PSE changed Igf1 promoter methylation in all three developmental stages, which was organ and sex specific. The PSE effect was less pronounced in adult offspring compared with the fetal and neonatal stages. In addition, the PSE effect in the adult stage was more pronounced in the lung compared with the liver. For most CpG sites, an inverse correlation was found for promoter methylation and mRNA expression when the data of all three stages were combined. This was more prominent in the liver. Our findings provide additional evidence for sex- and organ-dependent prenatal programming, which supports the developmental origins of health and disease (DOHaD) hypothesis.

Early-Life T-Helper 1 Immunity.

This brief review is written in memory of Eli Sercarz, a colleague who among many achievements, pioneered and revitalized early-life immunity, a field that is of high relevance to child health. For a long time, the neonatal stage was viewed as a window during which exposure to antigen (Ag) induces immune tolerance. In early 1990, however, it was discovered that the newborn mouse given Ag on the day of birth develops immunity when challenged later with the same Ag. But, these secondary responses displayed a deficit in T-helper (Th)1 cells and excess Th2 lymphocytes. Such discoveries explain the perceived tolerance of Ags given at the neonatal stage and correlate the paucity of effective neonatal vaccines and vulnerability to allergic reactions. Analyzing the mechanisms underlying neonatal Th1 cell deficits revealed a complex developmental interaction between Ag-presenting cells and the cytokines that they produced. This culminated into limited interleukin (IL)-12 in the environment and up-regulation of IL-13Rα1 expression and its association with IL-4Rα on the surface of Th1 cells. After Ag re-exposure, Th2 cells produce IL-4 and -13. Both bind to the heteroreceptor on Th1 cell surfaces and trigger their death. Usually, cytokines promote growth of T cells, but in this case IL-4 and -13 stimulate production of interferon regulatory factor 1 (IRF-1). IRF-1 translocates from nucleus to cytoplasm and stimulates apoptotic machinery that terminates Th1 cells. This suggests that vaccine formulations that could elevate IL-12 production are likely to counter IL-13Rα1 expression, preserve Th1 cells, and potentiate vaccine efficacy in neonates.

Effect of Stereotaxic Surgery of the Third Ventricle on Growth Performance in Neonatal Chicks.

Feeding behavior and energy metabolism are precisely regulated by humoral and/or neural factors in the central nervous system. In particular, nuclei, such as the arcuate nucleus, ventromedial hypothalamic nucleus, and lateral hypothalamic area located near the third ventricle of the hypothalamus are the centers of feeding and energy metabolism in various vertebrate species, including chickens. In this study, we evaluated the effects of cannulation of the third ventricle on chick growth and feeding behavior in the neonatal stage, to develop a method for local and chronic central nervous system-mediated energy metabolism. Referring to the chick brain atlas, a guide cannula was inserted into the third ventricle of the chick under anesthesia immediately after hatching using a stereotaxic instrument. The chicks that recovered from anesthesia were bred for 11 days under normal feeding management conditions, and then feed intake amount, body weight gain, and metabolic tissue weight were measured. The effects of direct stimulation of the third ventricle with 2-deoxy-D-glucose on the expression level of the immediate-early gene, cFOS, and feed intake in 5-day-old chicks were also evaluated. There were no differences in feed intake, body weight gain, and metabolic tissue weight between 11-day-old cannulated and control chicks. The expression of cFOS mRNA in the ventromedial hypothalamic nucleus was higher than that in the amygdala after the third ventricular administration of 2-deoxy-D-glucose. Additionally, direct third ventricular injection of 2-deoxy-D-glucose attenuated the feeding behavior of chicks for a while. Overall, we speculate that the technique is effective for local and/or chronic stimulation of the nucleus near the third ventricle of the chick hypothalamus, which is important for feed and energy metabolism regulation.

Vestibular evaluation in patients with congenital profound hearing loss using ice cold water caloric test and rotational chair test

Introduction: With the introduction of newborn hearing screening, hearing problems are often detected at the neonatal stage and in early infancy, and holistic approach including not only auditory but also vestibular function is needed. This study hopes to establish some corelation between cochlear and vestibular system dysfunction in a patient born with congenital profound hearing loss (CPHL). Materials and Methods: In this observational and descriptive study conducted at a tertiary care hospital, we evaluated eighty patients with bilateral CPHL from 2016 to 2018. After taking a detailed history and thorough ear, nose, and throat examination, the patients underwent vestibular function evaluation by ice cold water caloric test (ICCT) and rotational chair test (RCT) in the outpatient department, and the duration of nystagmus was calculated based on which hypoactive or absent vestibular function was found out. Results: In the present study, out of eighty patients with CPHL, vestibular function of 62 (77.50%) was normal, 9 (11.25%) hypoactive, and 9 (11.25%) with absent vestibular function. Conclusion: In the outpatient department where sophisticated equipment and laboratory facilities are not available, ICCT can be used to determine the vestibular function and also RCT can be used. From the present study, it can be concluded that vestibular dysfunction is present in 22.50% of patients with CPHL with the use of ICCT and RCT. There is a strong possibility of genetic origin etiology for vestibular dysfunction. The present study also concludes that consanguinity may be major etiological factor for vestibular dysfunction.

İzole Konjenital Kalp Hastalılarında Obstetrik ve Neonatal Sonuçlar - Tek Merkez Deneyimi

INTRODUCTION: Evidence shows that rather than postnatal diagnoses, prenatal diagnoses of congenital heart diseases are correlated with better long-term results and lower mortality rates. In this retrospective study, neonatal stage results of infants with isolated cardiac anomalies, their indications for operation or intervention (if any), and the mortality rates in the first age are attempted to be demonstrated. METHODS: The study was conducted through the retrospective analysis of 47 patients who were diagnosed with fetal cardiac anomaly in the antenatal period, and monitored during natal and postnatal stages at the same center in the Departments of Obstetrics and Gynecology and Pediatric Cardiology of Dokuz Eylul University Hospital. RESULTS: In our center 36 patients gave birth by cesarean section, and 11 patients by spontaneous vaginal delivery. Two low birthweight neonates were born (<2500 g), and these neonates were observed to have hypoplastic left heart and Fallot tetralogy. Thirty-nine of the 47 neonates born with isolated cardiac anomaly were operated after birth. Five patients were advised follow-ups. The remaining 3 neonates died within 48 hours after birth. Survival rate of the operated neonates was 59.5% and the general survival rate was 59.5%. Average 1, and 5-minute APGAR scores of the infants were 7.8 (3-9) and 9.0 (8-10), respectively The neonates were operated an average of 3 days after birth, and the average duration hospitalization of neonates were 12 days. Twenty-nine neonates needed preoperative respiratory support and 37 of them received prostaglandin E1 infusion. Follow-up was advised for 3 neonates within the first month and the first year. Five neonates needed operation during the specified period, and 2 neonates died. DISCUSSION AND CONCLUSION: In conclusion, prenatal diagnosis of congenital heart diseases allows planning the delivery in tertiary care centers, stabilization of the infants in the preoperative period and realization of emergency intervention.

Role of oxidative stress, genome damage and DNA methylation as determinants of pathological conditions in the newborn: an overview from conception to early neonatal stage

Increasing evidence suggests that early-life events can predispose the newborn to a variety of health issues in later life. In adverse pre- and perinatal conditions, oxidative stress appears to play an important role in the development of future pathological outcomes. From a molecular point of view, oxidative stress can result in genome damage and changes in DNA methylation that can in turn prime pathogenic mechanisms. Interestingly, both alterations have been related to a reciprocal regulation of oxidative stress. The aim of this review is to give a brief overview of the complex relationship linking oxidative stress to DNA damage and methylation and to go through the different sources of exposure that a neonate can encounter in utero or shortly after birth. In this context, the setup of methodologies to monitor the extent of oxidative stress, genomic damage and instability or the presence of altered methylation patterns contributes to the understanding on how the complex events occurring in early life can lead to either a healthy status or a pathological condition.

Oral administration of Debaryomyces hansenii CBS8339-β-glucan induces trained immunity in newborn goats

Beta-glucans from yeast can induce trained immunity in in vitro and in vivo models. Intraperitoneal doses of β-glucans in mammals have shown to induce trained immunity, but the training effects of orally administering β-glucans are unknown. Newborn goats are susceptible to infections in the neonatal stage, so the induction of trained immunity could improve animal survival. This study aimed to describe the in vitro effects of immunological training by β-glucan from Debaryomyces hansenii (β-Dh) on caprine monocytes, as well as its in vivo effects using oral doses on newborn goats upon challenge with lipopolysaccharide (LPS). Hence in vitro, goat monocytes trained with β-Dh up-regulated the gene expression of macrophage surface markers (CD11b and F4/80) whereas enhanced cell survival and high phagocytic ability was found upon LPS challenge. In the in vivo experiment, newborn goats stimulated with two doses (day −7 and - 4) of β-Dh (50 mg/kg) and challenged (day 0) with LPS showed an increase in respiratory burst activity, IL-1β, IL-6, and TNFα production in plasma, and transcription of the macrophage surface markers. This study has demonstrated for the first time that trained immunity was induced with oral doses of β-glucan upon LPS challenge in mammals using newborn goats.

Development of spermatogonial stem cell niche and immunoexpression of vimentin filaments in the testes of prenatal and postnatal Indian buffalo (Bubalusbubalis)

In this study, we established the components of the buffalo spermatogonial stem cell (SSC) niche in prenatal (38.5-280 days of gestation), neonatal, prepubertal, and adult testes histochemically and immunohistochemically. Immunohistochemically, the changes in vimentin expression were evaluated. During prenatal life, the niche of primordial germ cells was formed by the surrounding mesenchymal cells and fibroblasts. The basement membrane (BM) was not organized at this stage. At 7.6 cm crown-rump length, testicular cords with a thin BM were seen. At the late gestational period, testicular cords with well-organized BM were seen. At the neonatal stage, the testicular cords were lined with distinct BM, while during the prepubertal period, distinct BM with peritubular myoid cells (PMCs) were seen. In adult testes, the seminiferous tubules were surrounded by a thick BM surrounded by PMCs. PM cells were only single-layered as seen in H&E-stained paraffin sections, picrosirius red-stained, and Masson?s trichrome-stained paraffin sections. Collagen fibers were seen to increase quantitatively to provide strength and cushioning to the developing niche of spermatogonial stem cells in the testes. The expression of vimentin increased to a certain extent until adult life, indicating its significant role as an intermediate filament during development.

Stomach content and stable isotopes reveal an ontogenetic dietary shift of young-of-the-year scalloped hammerhead sharks (Sphyrna lewini) inhabiting coastal nursery areas

Sphyrna lewini is a placental viviparous shark that uses coastal nursery areas in Jalisco, Mexico, where pups stay 4–12 months. Changes in size or swimming speed may be reflected in diet composition. The main objectives of this study were as follows: (1) analyze the differences in trophic ecology of juvenile S. lewini from coastal nursery areas of Jalisco by sex and size, through stomach content and stable isotope analyses; and (2) analyze changes in muscle and liver δ15N and δ13C values with shark size. Samples were collected from the artisanal fishery from September 2013 to December 2016. Three size classes were compared: neonates, stretched total length (STL) ≤ 75 cm, and STL > 75 cm (75–100 cm). Bony fishes were the most important group in all size classes, and the importance of shrimps decreased with STL. Significant differences in diet composition were found between neonates and STL > 75 cm, which showed the lowest niche overlap (0.32). STL ≤ 75 cm showed the largest niche width (0.75). The δ15N muscle and liver values declined with total length, reflecting the maternal isotopic signal. No differences in liver δ13C values were observed among size classes, but larger size class showed higher muscle δ13C values. Both isotope and stomach content analyses classified all sharks as tertiary consumers, but the trophic position (TP) estimated using δ15N was higher in neonates. No significant differences in the diet and TP were observed between sexes. The liver C:N ratio decreased sharply with STL up to 55 cm, from which increased smoothly, reflecting the lipid reserves consumed during their neonatal stage.

Epidemiological analysis of neonatal CHD near Petrochemical Complex.

To study the incidence, types, and characteristics of CHD in all live births delivered in Jinshan Hospital from 1 January, 2016 to 31 December, 2017, and to analyse the correlation between CHD and the distance between maternal residence and Shanghai Petrochemical Complex. All live births, delivered in Jinshan Hospital in 2016 and 2017, have received CHD screening and neonatal follow-up after delivery. For those both positive for CHD screening and negative for CHD screening but with heart murmur found during physical examination on neonatal follow-up after delivery, echocardiography was performed to confirm CHD diagnosis. All maternal residential addresses have been grouped for analysis according to the distance between maternal residence and Shanghai Petrochemical Complex. There were 5544 live births in total, and a total of 79 children with CHD were confirmed by CHD screening and echocardiography, of which the types of diseases with high incidence, in descending order of incidence, are atrial septal defect (48/79), ventricular septal defect (25/79), patent ductus arteriosus (21/79), and pulmonary artery stenosis (9/79). There was no statistically significant difference in the incidence of CHD among the groups divided by the distance between maternal residence and Shanghai Petrochemical Complex. The incidence of neonatal CHD near Shanghai Petrochemical Complex is 1.42%. Most of the children with CHD can be screened out through CHD screening and physical examination in neonatal period and early infancy stage. The distance between maternal residence and Shanghai Petrochemical Complex has no significant direct effect on the incidence of CHD in neonates near Shanghai Petrochemical Complex.

Thyroxine modulation of immune toxicity induced by mixture pesticides mancozeb and fipronil in mice

AimThe cross regulation between neuroendocrine system, particularly Hypothalamus-Pituitary-Thyroid (HPT) axis and immune system during embryonic/early neonatal developmental stages shapes the functional attribute of immune response throughout the life. Thus, disruption of immune system was anticipated on exposure to thyroid disrupting pesticides (TDPs) mancozeb (MCZ) and fipronil (FPN) during critical windows of early postnatal days (PND) development. Main methodsMice were exposed to MCZ and FPN as individual (0.5% LD 50 each) and as mixtures (0.25% and 0.5% LD 50 each) from PND 31 (initiation phase of immune response) till PND 60 (Maturation phase). Thyroxine (T4) supplementation was given from PND 51 to PND 60. Assessment was done at PND 61 as well as at PND 91 (adults). Key findingsPlasma level of thyroid hormones (T3 and T4) was reduced but pituitary hormone (TSH) increased till adulthood on exposure to mixture pesticides but not on individual exposure. Mixture pesticides also increased body weight gain and reduced survival rate in adults. Exposure of individual pesticides exert immunotoxicity but more pronounced immune suppression was observed in mixture pesticides exposed group as reflected in reduced relative weight and cellularity in spleen and thymus, reduced in vitro mitogenic (Con A/LPS) response of splenocytes and thymocytes (reduced proliferative index and increased apoptotic/necrotic death). T4 supplementation ameliorated thyroid disruptive and immunotoxic effect of pesticides. SignificanceThe additive/synergistic toxicity as well as hypothyroidism induced by mixture pesticides has produced pronounced immune suppression that reflected till adulthood. Supplementation of T4 prevented thyroid axis disruption mediated immunosuppression.

Fetal and neonatal genistein exposure aggravates to interfere with ovarian follicle development of obese female mice induced by high-fat diet

With epidemic of obesity, it affects aspects of female reproduction. Genistein could ameliorate obesity in people and animals, but might exert adverse effects on the female reproductive system. To evaluate the effects of fetal and neonatal genistein exposure on the ovarian health of F1 obese female mice with obesity induced by high-fat diet after weaning, we simulated a diet-induced obesity model to observe and determine biological effects of genistein exposure on the ovarian follicle of overfed female mice. Results showed that F1 female mice with obesity induced by high-fat diet significantly prolonged the estrus cycle, disrupted sex hormonal balance and ovarian follicle development after they were exposed to 25 mg/kg b.w./day of genistein during the fetal and neonatal stages. Genistein significantly up-regulated the ovarian mRNA expression of estrogen receptor beta in F1 obese female mice, and high-fat diet influenced the ovarian mRNA expression of estrogen receptor alpha, luteinizing hormone receptor and follicle-stimulating hormone receptor. Hence, genistein exposure from the fetal stage might increase the risk of reproductive diseases in obese females in later life. Thus, the long-term risks of genistein to obese females should be thoroughly assessed.

Didelphis albiventris: an overview of unprecedented transcriptome sequencing of the white-eared opossum

BackgroundThe white-eared opossum (Didelphis albiventris) is widely distributed throughout Brazil and South America. It has been used as an animal model for studying different scientific questions ranging from the restoration of degraded green areas to medical aspects of Chagas disease, leishmaniasis and resistance against snake venom. As a marsupial, D. albiventris can also contribute to the understanding of the molecular mechanisms that govern the different stages of organogenesis. Opossum joeys are born after only 13 days, and the final stages of organogenesis occur when the neonates are inside the pouch, depending on lactation. As neither the genome of this opossum species nor its transcriptome has been completely sequenced, the use of D. albiventris as an animal model is limited. In this work, we sequenced the D. albiventris transcriptome by RNA-seq to obtain the first catalogue of differentially expressed (DE) genes and gene ontology (GO) annotations during the neonatal stages of marsupial development.ResultsThe D. albiventris transcriptome was obtained from whole neonates harvested at birth (P0), at 5 days of age (P5) and at 10 days of age (P10). The de novo assembly of these transcripts generated 85,338 transcripts. Approximately 30% of these transcripts could be mapped against the amino acid sequences of M. domestica, the evolutionarily closest relative of D. albiventris to be sequenced thus far. Among the expressed transcripts, 2077 were found to be DE between P0 and P5, 13,780 between P0 and P10, and 1453 between P5 and P10. The enriched GO terms were mainly related to the immune system, blood tissue development and differentiation, vision, hearing, digestion, the CNS and limb development.ConclusionsThe elucidation of opossum transcriptomes provides an out-group for better understanding the distinct characteristics associated with the evolution of mammalian species. This study provides the first transcriptome sequences and catalogue of genes for a marsupial species at different neonatal stages, allowing the study of the mechanisms involved in organogenesis.

Cortical Foxp2 Supports Behavioral Flexibility and Developmental Dopamine D1 Receptor Expression.

Genetic studies have associated FOXP2 variation with speech and language disorders and other neurodevelopmental disorders (NDDs) involving pathology of the cortex. In this brain region, FoxP2 is expressed from development into adulthood, but little is known about its downstream molecular and behavioral functions. Here, we characterized cortex-specific Foxp2 conditional knockout mice and found a major deficit in reversal learning, a form of behavioral flexibility. In contrast, they showed normal activity levels, anxiety, and vocalizations, save for a slight decrease in neonatal call loudness. These behavioral phenotypes were accompanied by decreased cortical dopamine D1 receptor (D1R) expression at neonatal and adult stages, while general cortical development remained unaffected. Finally, using single-cell transcriptomics, we identified at least five excitatory and three inhibitory D1R-expressing cell types in neonatal frontal cortex, and we found changes in D1R cell type composition and gene expression upon cortical Foxp2 deletion. Strikingly, these alterations included non-cell-autonomous changes in upper layer neurons and interneurons. Together, these data support a role for Foxp2 in the development of dopamine-modulated cortical circuits and behaviors relevant to NDDs.

Myocardial overexpression of ANKRD1 causes sinus venosus defects and progressive diastolic dysfunction

Increased Ankyrin Repeat Domain 1 (ANKRD1) levels linked to gain of function mutations have been associated to total anomalous pulmonary venous return and adult cardiomyopathy occurrence in humans. The link between increased ANKRD1 level and cardiac structural and functional disease is not understood. To get insight into this problem, we have generated a gain of function ANKRD1 mouse model by overexpressing ANKRD1 in the myocardium. Ankrd1 is expressed non-homogeneously in the embryonic myocardium, with a dynamic nucleo-sarcomeric localization in developing cardiomyocytes. ANKRD1 transgenic mice present sinus venosus defect, which originates during development by impaired remodelling of early embryonic heart. Adult transgenic hearts develop diastolic dysfunction with preserved ejection fraction, which progressively evolves into heart failure, as shown histologically and haemodynamically. Transgenic cardiomyocyte structure, sarcomeric assembly, and stability are progressively impaired from embryonic to adult life. Postnatal transgenic myofibrils also present characteristic functional alterations: impaired compliance at neonatal stage and impaired lusitropism in adult hearts. Altogether, our combined analyses suggest that impaired embryonic remodelling and adult heart dysfunction in ANKRD1 transgenic mice present a common ground of initial cardiomyocyte defects, which are exacerbated postnatally. Molecular analysis showed transient activation of GATA4-Nkx2.5 transcription in early transgenic embryos and subsequent dynamic transcriptional modulation within titin gene. ANKRD1 is a fine mediator of cardiomyocyte response to haemodynamic load in the developing and adult heart. Increased ANKRD1 levels are sufficient to initiate an altered cellular phenotype, which is progressively exacerbated into a pathological organ response by the high ventricular workload during postnatal life. Our study defines for the first time a unifying picture for ANKRD1 role in heart development and disease and provides the first mechanistic link between ANKRD1 overexpression and cardiac disease onset.

Homeostatic Function of Dermokine in the Skin Barrier and Inflammation

Dermokine is a chiefly skin-specific secreted glycoprotein localized in the upper epidermis, and its family consists of three splice variants in mice and five in humans. To investigate the pathophysiological impact of dermokine, we generated mice deficient for two (βγ) or all dermokine isoforms (αβγ). Both variants, especially dermokine αβγ-deficient mice exhibited scale and wrinkle formation resembling ichthyosis accompanied by transepidermal water imbalance at the neonatal stage. Several dermokine αβγ-deficient mice died by postnatal day 21 when reared under low humidity. Moreover, the cornified envelope was vulnerable, and skin barrier lipid ceramides were reduced in the epidermis of dermokine αβγ-deficient mice. cDNA microarray and quantitative reverse transcriptase-PCR assays of the epidermis revealed the upregulation of small proline-rich protein and late cornified envelope family members, as well as antimicrobial peptides in the dermokine αβγ-deficient mice. These barrier gene signatures were similar to that seen in psoriasis, whereas recent studies demonstrated that congenital ichthyosis has gene profiles resembling psoriasis. In line with these findings, adult dermokine αβγ-deficient mice exhibited aggravated phenotypes in psoriasis-like dermatitis models but not in allergic dermatitis models. Dermokine may play a regulatory role in inflammatory dyskeratotic diseases, such as congenital ichthyosis and psoriasis, in the crosstalk between barrier dysfunction and inflammation.

Early Development of Resident Macrophages in the Mouse Cochlea Depends on Yolk Sac Hematopoiesis.

Resident macrophages reside in all tissues throughout the body and play a central role in both tissue homeostasis and inflammation. Although the inner ear was once believed to be “immune-privileged,” recent studies have shown that macrophages are distributed in the cochlea and may play important roles in the immune system thereof. Resident macrophages have heterogeneous origins among tissues and throughout developmental stages. However, the origins of embryonic cochlear macrophages remain unknown. Here, we show that the early development of resident macrophages in the mouse cochlea depends on yolk sac hematopoiesis. Accordingly, our results found that macrophages emerging around the developing otocyst at E10.5 exhibited dynamic changes in distribution and in situ proliferative capacity during embryonic and neonatal stages. Cochlear examination in Csf1r-null mice revealed a substantial decrease in the number of Iba1-positive macrophages in the spiral ganglion and spiral ligament, whereas they were still observed in the cochlear mesenchyme or on the intraluminal surface of the perilymphatic space. Our results demonstrated that two subtypes of resident macrophages are present in the embryonic cochlea, one being Csf1r-dependent macrophages that originate from the yolk sac and the other being Csf1r-independent macrophages that appear to be derived from the fetal liver via systemic circulation. We consider the present study to be a starting point for elucidating the roles of embryonic cochlear resident macrophages. Furthermore, resident macrophages in the embryonic cochlea could be a novel target for the treatment of various inner ear disorders.

Early skeletal muscle pathology and disease progress in the dy3K/dy3K mouse model of congenital muscular dystrophy with laminin \u03b12 chain-deficiency

Deficiency of laminin α2 chain leads to a severe form of congenital muscular dystrophy (LAMA2-CMD), and dystrophic symptoms progress rapidly in early childhood. Currently, there is no treatment for this detrimental disorder. Development of therapies is largely hindered by lack of understanding of mechanisms involved in the disease initiation and progress, both in patients but also in mouse models that are commonly used in the preclinical setup. Here, we unveil the first pathogenic events and characterise the disease development in a mouse model for LAMA2-CMD (dy3K/dy3K), by analysing muscles at perinatal, neonatal and postnatal stages. We found that apoptotic muscle fibres were present as early as postnatal day 1. Other typical dystrophic hallmarks (muscle degeneration, inflammation, and extensive production of the extracellular matrix proteins) were clearly evident already at postnatal day 4, and the highest degree of muscle deterioration was reached by day 7. Interestingly, the severe phenotype of limb muscles partially recovered on days 14 and 21, despite worsening of the general condition of the dy3K/dy3K mouse by that age. We found that masticatory muscles were severely affected in dy3K/dy3K mice and this may be an underlying cause of their malnutrition, which contributes to death around day 21. We also showed that several signalling pathways were affected already in 1-day-old dy3K/dy3K muscle. Therapeutic tests in the dy3K/dy3K mouse model should therefore be initiated shortly after birth, but should also take into account timing and correlation between regenerative and pathogenic events.

Decomposition of a Multiscale Entropy Tensor for Sleep Stage Identification in Preterm Infants

Established sleep cycling is one of the main hallmarks of early brain development in preterm infants, therefore, automated classification of the sleep stages in preterm infants can be used to assess the neonate’s cerebral maturation. Tensor algebra is a powerful tool to analyze multidimensional data and has proven successful in many applications. In this paper, a novel unsupervised algorithm to identify neonatal sleep stages based on the decomposition of a multiscale entropy tensor is presented. The method relies on the difference in electroencephalography(EEG) complexity between the neonatal sleep stages and is evaluated on a dataset of 97 EEG recordings. An average sensitivity, specificity, accuracy and area under the receiver operating characteristic curve of 0.80, 0.79, 0.79 and 0.87 was obtained if the rank of the tensor decomposition is selected based on the age of the infant.

348 Homeostatic functions of dermokine in skin barrier and innate immunity

Dermokine (DMKN) is a secreted glycoprotein and its family members consist of five splicing variants (α, β, γ, δ and ε)in human and three variants (α, β and γ) in mice. Especially, α, β and γ isoforms are expressed in the upper epidermis. DMKN α represents the C-terminal end of β isoform, which is responsible for cytokine-like action and inhibits phosphorylation of ERK, leading to suppression of aberrant keratinocyte proliferation in vitro. To investigate the DMKN function in vivo, we generated mice deficient for all the three DMKN α, β and γ. The mice exhibited dry scaly skinand deep wrinkle formation with tail constriction and autoamputation in the neonatal stage, contributing to increased TEWL. Some of them showed growth retardation and death at low humidity. The skin pathology revealed thickening of corneal and granular layers with increased proteolytic activity in the corneal layers and cornified envelope fragility. cDNA microarray and quantitative RT-PCR assays using the mouse epidermis disclosed upregulation of small proline-rich protein and late CE family members, clustering genes for CE assembly, in parallel with increased levels of antimicrobial peptides. DMKN deficiency did not affect the development of DNFB- or FITC-induced contact hypersensitivity models, as well as OVA- or MC903-induced atopic dermatitis models in adult mice. However, DMKN-deficient mice showed exacerbation of psoriasiform dermatitis models induced by topical imiquimod application or intradermal IL-23 injection. Augmented expression profile of TNF-α/IL-17/IL-23 and ELR+ CXC chemokines were characteristic for psoriasis. Thus, DMKN possesses dual biological properties for skin barrier maintenance and regulatory innate immune response associated with psoriasis.