Whole-exome sequencing for genetic diagnosis of idiopathic liver injury in children.

Genome-wide approaches, such as whole-exome sequencing (WES), are widely used to decipher the genetic mechanisms underlying inter-individual variability in disease susceptibility. We aimed to dissect inborn monogenic determinants of idiopathic liver injury in otherwise healthy children. We thus performed WES for 20 patients presented with paediatric-onset recurrent elevated transaminases (rELT) or acute liver failure (ALF) of unknown aetiology. A stringent variant screening was undertaken on a manually-curated panel of 380 genes predisposing to inherited human diseases with hepatobiliary involvement in the OMIM database. We identified rare nonsynonymous variants in nine genes in six patients (five rELT and one ALF). We next performed a case-level evaluation to assess the causal concordance between the gene mutated and clinical symptoms of the affected patient. A genetic diagnosis was confirmed in four rELT patients (40%), among whom two carried novel mutations in ACOX2 or PYGL, and two had previously-reported morbid variants in ABCB4 or PHKA2. We also detected rare variants with uncertain clinical significance in CDAN1, JAG1, PCK2, SLC27A5 or VPS33B in rELT or ALF patients. In conclusion, implementation of WES improves diagnostic yield and enables precision management in paediatric cases of liver injury with unknown aetiology, in particular recurrent hypertransaminasemia.

Evaluation and 3D imaging of mineral structure changes of the rabbit (New Zealand) skull during developmental periods.

This study aimed to determine the morphometric measurements anatomically and CT images of skulls of healthy male and female rabbits during postnatal development, to analyse the data statistically and to demonstrate the structural changes in bone. A total of 40 rabbits (20 females and 20 males) were divided into four groups including prepubertal period (group I (0-1 month)), period between adolescence and adulthood (group II (3-5 month)) and later (young adult period as group III (1-3 years) and old adult period as group IV (3-5 years)), with five animals in each group. After the morphometric measurements, the surface area and volume values of the skull were calculated. The skulls were reconstructed using a 3D Slicer (5.0.2), which is used for 3D modelling. The cranial bones in each group were then crushed using a grinder so that the powdered samples were obtained for XRF (X-ray fluorescence technique). The p-value was statistically highly significant between group and gender (p < 0.001). In morphometric measurements, males were generally higher than females. Only PL, GBOC and GNB measurements were higher in females. The p-value between groups (in all measurements), between genders (in TL, GLN, FL, VL, OZB and GBN parameters) and between groups and genders (in TL, DL and VL parameters) was statistically highly significant (p < 0.001). The p-value between the groups, p-value between sexes and p-value between group and sex in Si, P, K, Ca, Ni, Zn, Sr, Sr and Ca/P elements were statistically significant (p < 0.001). Consequently, metric, volume and surface area measurements were taken through 3D modelling of skull bone in prepubertal period (group I), period between adolescence and adulthood (group II) and later (young adult period as group III and old adult period as group IV) of rabbits and the change in the mineral structure during postnatal development and effect of sex on this change were investigated. This might be the first study to assess both metric and mineral changes at four age intervals taken during the life span of rabbits.

Analysis and 3D Imaging of Morphometric - Mineral Structure Changes in Skulls of Male and Female Guinea Pigs During and After Developmental Period

SUMMARY: Even though morphometric and mineral studies related to the guinea pig (Cavia porcellus) skull have been carried out, this study is the first attempt to evaluate all developmental stages of male and female guinea pigs. This study aims to this study is to create 3D modeling of CT images obtained from the skulls of male and female guinea pigs during the developmental period (prepuberty and the period between puberty and adulthood) and following periods (young adulthood and old adulthood), to analyze some biometric bone data such as volume, surface area and length, and to assess the developmental analysis of the mineral matter change in their skulls. The CT-scanned skulls were transferred to 3D Slicer (5.0.2), which is used for 3D modeling. The surface area and volume were calculated by measuring the measurement points on the models. In addition, the XRF device was used to show elemental ratio changes during different developmental stages. According to metric measurements, a gradual increase was observed during the life period. The metric measurements of the skull bone had a higher measurement value in male guinea pigs than in their female counterparts. While Ca/P ratio increased up to the third group and partially decreased in the fourth group in males, it gradually increased from the first group to the fourth group in females. This study revealed that puberty, adulthood and sex were effective in the physical and chemical characterization of skull bone structure in guinea pigs.

Aunque se han realizado estudios morfométricos y de minerales relacionados con el cráneo del cobayo (Cavia porcellus), esta investigación es el primer intento de evaluar las etapas de desarrollo de cobayos machos y hembras. El objetivo de este estudio fue crear un modelado 3D de imágenes de tomografía computarizada obtenidas de los cráneos de cobayos machos y hembras durante el período de desarrollo (prepubertad y el período entre la pubertad y la edad adulta) y los períodos siguientes (edad adulta joven y edad adulta mayor), para analizar algunos datos biométricos de los huesos, como el volumen, la superficie y la longitud, y además, analizar el cambio de materia mineral en sus cráneos durante el desarrollo. Los cráneos escaneados se transfirieron a 3D Slicer (5.0.2), que se utiliza para el modelado 3D. El área de superficie y el volumen se calcularon midiendo los puntos de medición en los modelos. Además, se utilizó el dispositivo XRF para mostrar los cambios en las proporciones elementales durante diferentes etapas de desarrollo. Según mediciones métricas, se observó un aumento gradual durante el período de vida. Las medidas métricas del hueso del cráneo tuvieron un valor de medición más alto en los cobayos machos que en las hembras. Mientras que la relación Ca/P aumentó hasta el tercer grupo y disminuyó parcialmente en el cuarto grupo en los machos y aumentó gradualmente del primer grupo al cuarto grupo en las hembras. Este estudio reveló que la pubertad, la edad adulta y el sexo fueron efectivos en la caracterización física y química de la estructura ósea del cráneo en cobayos.

Activities of the hypothalamo-pituitary-adrenal axis and autonomic nervous system following a strong earthquake.

This study aimed to investigate the effects of post-traumatic stress, caused by a strong earthquake, on the hypothalamo-pituitary-adrenal axis (HPA) and autonomous nervous system activity (ANS). Activities of the HPA (as salivary cortisol) and ANS (as heart-rate variability [HRV]) were measured following the 2020 Elazig (Türkiye) earthquake (6.8 Richter Scale, classified as strong). A total of 227 participants (103 men (45%) and 124 women (%55)) provided saliva samples twice, namely, 1 week and 6 weeks after the earthquake. Of these participants, HRV was measured in 51 participants by 5 min continuous electrocardiogram (ECG) recording. Frequency- and time-domain parameters of the HRV were calculated to assess the activity of ANS and low/high frequency (LF/HF) ratio was used as surrogate for sympathovagal balance. Salivary cortisol levels decreased from week 1 towards week 6 (17.40 ± 1.48 and 15.32 ± 1.37 ng/mL, respectively, p < 0.05). There were no gender differences (17.99 ± 2.63 and 16.90 ± 1.60 ng/mL, respectively for females and males, p > 0.05) for salivary cortisol levels. There were no differences in time- and frequency domain parameters of the HRV including LF/HF ratio (2.95 ± 0.38 ms2 and 3.60 ± 0.70 ms2 , respectively for week 1 and 6, p > 0.05). The data show that HPA axis activity, but not that of the ANS, remains higher 1 week after the earthquake but decreases afterwards towards the sixth week, suggesting that the HPA axis might be responsible for the long-term effects of a traumatic event like a strong earthquake.

Physical and chemical characterization of the femur during and after the body development period in male and female guinea pigs.

In this study, it was aimed to reveal the physical and chemical characterization of the bone structures during body development periods (prepubertal period, period between adolescence and adulthood) and after (young adult period and old adult period) in male and female guinea pigs. In this study, 40 guinea pigs (20 male, 20 female) were used. Morphometric measurements, X-ray fluorescence (XRF) analysis for mineral levels, Brunauer-Emmett-Teller (BET) analysis for surface area, and porosity analysis were applied to the bones. The male guinea pigs had greater values than females in the other three categories, with the exception of the second group, when the females have higher values in morphometric measurements. Ca levels rose up to the third group, as did P levels in the males, peaking in the third group and declining in the fourth. As with phosphorus, there was a progressive rise in females from the first to the fourth group. Fe, Zn, and Sr elements had the greatest values in both genders in the first group. In all four groups, the females had greater Zn levels than males. The highest Ca/P ratio was found in the third male group and the fourth female group. This study revealed that adolescence, adulthood, and gender are effective in the physical and chemical characterization of bone structure in guinea pigs.

A small non-interface surface epitope in human IL18 mediates the dynamics and self-assembly of IL18-IL18BP heterodimers.

Interleukin 18 (IL18) is a pro-inflammatory cytokine that modulates innate and adaptive immune responses. IL18 activity is tightly controlled by the constitutively secreted IL18 binding protein (IL18BP). PDB structures of human IL18 showed that a short stretch of amino acids between 68 and 81 adopted a disordered conformation in all IL18-IL18BP complexes while adopting a 310 helical structure in other IL18 structures, including the receptor complexes. The C74 of human IL18, which was reported to form a novel intermolecular disulfide bond in the human tetrameric assembly, is also located in this short epitope. These observations reflected the importance of this short surface epitope for the structure and dynamics of the IL18-IL18BP heterodimers. We have analyzed all known IL18-IL18BP complexes in the PDB by all-atom MD simulations. The analysis also included two computed complex models adopting a helical structure for the surface epitope. Heterodimer simulations showed a stabilizing impact of the small surface region at the helical form by reducing flexibility of the complex backbone. Analysis of the symmetry-related human IL18-IL18BP tetramer showed that the unfolding of this small surface region also contributed to the IL18-IL18BP stability through a completely exposed C74 sidechain to form an intermolecular disulfide bond in the self-assembled human IL18-IL18BP dimer. Our findings showed how the conformation of the short IL18 epitope between amino acids 68 and 81 would affect IL18 activity by mediating the intermolecular interactions of IL18.

Physical and chemical characterization of femur during and after body development period in male and female rats.

The aim of this study was to determine the physical and chemical characteristics of the bone structures during four developmental periods. Forty Wistar Albino rats (20 male and 20 female) were divided into four groups including prepubertal period (group I), period between adolescence and adulthood (group II) and later (young adult period as group III and old adult period as group IV). The bones were analysed by morphometric measurements, XRF (X-ray fluorescence) analysis for mineral levels and BET analysis (Brunaurer-Emmett-Teller) for surface area and porosity. In morphometric measurements, the GL (greatest length) and the GLC (greatest length from caput femoris) values increased gradually from the first to the fourth group, and these values were higher in the males than the females. Phosphorus and calcium values were higher in the males in adult groups (third and fourth) compared to that for the females; however, they were higher in the females in groups up to adulthood (first and second). While the Ca/P ratio reached the highest value in the second group in the male, it decreased gradually afterwards. In females, the rate, which was close to each other in the first three groups, increased in the fourth group. Surface area size in the female and the male rats was the highest in the second group. In conclusion, changes in the rat bone structure during the development and adulthood periods of the body were revealed, and it was determined that the gender factor was effective in these changes.

Inherited IRAK-4 Deficiency in Acute Human Herpesvirus-6 Encephalitis.

Human herpesvirus-6 (HHV-6) infection can rarely cause life-threatening conditions, such as encephalitis, in otherwise healthy children, with unclear pathogenesis. We studied a child who presented with acute HHV-6 encephalitis at the age of 10 months and who was homozygous for a novel missense mutation in IRAK4, encoding interleukin-1 receptor-associated kinase 4, identified by whole-exome sequencing. We tested the damaging impact of this mutation in silico by molecular dynamics simulations and in vitro by biochemical and functional experiments utilizing cell lines and patient's cells. We found that the mutation is severely hypomorphic, impairing both the expression and function of IRAK-4. Patient's leukocytes had barely detectable levels of IRAK-4 and diminished anti-viral immune responses to various stimuli inducing different Toll-like receptors and cytosolic nucleic acid sensors. Overall, these findings suggest that acute HHV-6 encephalitis can result from inborn errors of immunity to virus. This study represents the first report of isolated acute HHV-6 infection causing encephalitis in an inherited primary immunodeficiency, notably autosomal recessive (AR) partial IRAK-4 deficiency, and the first report of AR IRAK-4 deficiency presenting with a severe viral disease, notably HHV-6 encephalitis upon an acute infection, thereby expanding the clinical spectrum of IRAK-4 deficiency.

Investigation of the effect of training on serotonin, melatonin and hematologic parameters in adolescent basketball players.

OBJECTIVES: Exercise can improve both health and mood. Some beneficial effects of exercise are attributed to endocrine status. This study aims to evaluate the effect of eight weeks of basketball training on melatonin, serotonin, and hematologic parameters in basketball players. METHODS: The experimental group was selected form 34 healthy young boys, aged between 13 and 16 years old. The participants were randomly assigned to the control group (n=17) and the exercise group (n=17). The exercise program consisted of 2 h/day aerobic activity of basketball training in 5 days a week for 8 weeks. Venous blood was taken on the day before experiment (pre-exercise) and on the day following the last exercise (post-exercise) and hormone levels were detected by ELISA. RESULTS: Serotonin and melatonin levels significantly increased in the post-exercise group compared to the other groups (p<0.05). Exercise caused increase in WBC, RBC, HCT and Hb levels (p<0.05) while did not alter PLT, MCH, and PCT levels (p>0.05). This study indicates that an eight weeks-long regular aerobic exercise increased melatonin and serotonin levels, and also altered some hematological parameters. CONCLUSIONS: In conclusion, it is believed that improvement in levels of serotonin, melatonin, and hematological parameters after eight weeks of regular basketball training in basketball players could be attributed to beneficial effects of exercise. Investigation in other branches of sports and in different gender and age groups would make contribution into exercise physiology and training science.