Skeletal Malformations Research Articles

PATHOLOGY OF HATCHLING HAWKSBILL SEA TURTLE (ERETMOCHELYS IMBRICATA) MORTALITIES OCCURRING WHILE UNDER REHABILITATIVE CARE, 2015-21.

Increasing hatchling survival is an important element of conservation of the critically endangered hawksbill sea turtle (Eretmochelys imbricata). Yet, there is little information regarding mortality-associated pathological states of hawksbill hatchlings. The aim of this study was to describe lesions affecting hawksbill hatchlings that died while under rehabilitative care. Forty-four turtles representing the nesting sites of two islands and a 7-yr study period were subjected to comprehensive postmortem examination. The most common lesions included dermatitis (34%), skeletal malformations (23%), and pneumonia (23%). Dermatitis and pneumonia were caused by a variety of presumptively opportunistic bacterial and fungal infections. Fungal infections affected 23% of study turtles, also causing rhinitis and esophagitis. Around half of the cases of dermatitis presented with history of skin lesions, and all those involving periocular areas had clinical history of eye lesions. Pneumonia was not predicted by clinical signs or time in rehabilitation. Malformations included carapace compressions, supra- or subnumerary scutes, and dysmelias with many of those affected having concurrent pathology involving other organs. Other lesions included bacterial yolk sacculitis (15%), skeletal muscle degeneration and necrosis (13%), and acute renal tubular necrosis (13%). The study population was female biased (93%), raising concern for skewed hatchling sex ratios and high incubation temperatures in the eastern Caribbean. The pathology described by this study improves our understanding of threats to hawksbill hatchlings and may be taken into consideration by clinicians when implementing strategies for rehabilitative care.

Heterozygous variant in FGFR3 underlying severe phenotypes in the second trimester: a case report

BackgroundAchondroplasia is a congenital skeletal system malformation caused by missense variant of FGFR3 gene with an incidence of 1 per 20,000–30,000 newborns, which is an autosomal dominant inheritance disease. Despite similar imaging features, the homozygous achondroplasia is absolutely lethal due to thoracic stenosis, whereas heterozygous achondroplasia does not lead to fetal death.Case presentationA fetus with progressive rhizomelic short limbs and overt narrow chest was detected by prenatal ultrasound in the second trimester. Gene sequencing results of amniotic fluid sample indicated a rare missense variant NM_000142.4: c.1123G > T(p.Gly375Cys), leading to a glycine to cysteine substitution. Re-sequencing confirmed that it was a heterozygous variant, and thoracic stenosis was then confirmed in the corpse by radiological examination.ConclusionsWe identified a heterozygous variant of the FGFR3 gene as the rare pathogenic variant of severe achondroplasia in a fetus. Heterozygous variants of p.Gly375Cys may have a severe phenotype similar to homozygote. It’s crucial to combine prenatal ultrasound with genetic examination to differentiate heterozygous from homozygous achondroplasia. The p.Gly375Cys variant of FGFR3 gene may serve as a vital target for the diagnosis of severe achondroplasia.

Unique clinical presentations and follow-up outcomes from experience with congenital disorders of glycosylation: PMM2-PGM1-DPAGT1-MPI-POMT2-B3GALNT2-DPM1-SRD5A3-CDG.

Congenital Glycosylation Disorders (CDG) are a large group of inherited metabolic diseases with multi-organ involvement. Herein, we aimed to expand the clinical characteristics of patients with CDG based on our experience with diagnoses and follow-up of CDG patients from different subtypes. The clinical and laboratory findings from the last 15 years were reviewed retrospectively in Ege University Child Metabolism and Nutrition Department. There were 8 (57.2 %) females and 6 (42.8 %) males. Diagnoses of the patients were PMM2-CDG (n=4), PGM1-CDG (n=2), DPAGT1-CDG (n=2), SRD5A3-CDG (n=2), MPI-CDG (n=1), POMT2-CDG (n=1), B3GALNT2-CDG (n=1), DPM1-CDG (n=1). The clinical findings of the patients were dysmorphia (85.7 %), developmental delay (85.7 %), intellectual disability (85.7 %), ocular abnormalities (64.2 %), skeletal malformations (64.2 %), failure to thrive (57.1 %), microcephaly (57.1 %), hepatomegaly (35.7 %), hearing loss (35.7 %), seizures (28.5 %), gastrointestinal symptoms (21.4 %), endocrine abnormalities (21.4 %), and cardiac abnormalities (7.1 %). Laboratory findings were abnormal TIEF (92.8 %), abnormal liver enzymes (64.2 %), decreased protein C (64.2 %), decreased antithrombin III (64.2 %), decreased protein S (42.8 %), hypogammaglobulinemia (35.7 %), cerebellar hypoplasia (28.5 %), CK elevation (7.1 %), and hypoglycemia (7.1 %). This study contributes to the literature by sharing our ultra-rare DPM1-CDG case with less than 20 cases in the literature and expanding the clinical and molecular characteristics of other CDG patients. Hyperinsulinemic hypoglycemia, short stature, hypothyroidism, growth hormone deficiency, hypogammaglobulinemia, pericardial effusion, elevated CK, congenital myasthenia, and anorectal malformation were unique findings that were observed. Cerebello-ocular findings accompanying multi-organ involvement were an essential clue for a possible CDG.

Autoimmunity and immunodeficiency associated with monoallelic LIG4 mutations via haploinsufficiency

Biallelic mutations in LIG4 encoding DNA-ligase 4 cause a rare immunodeficiency syndrome manifesting as infant-onset life-threatening and/or opportunistic infections, skeletal malformations, radiosensitivity and neoplasia. LIG4 is pivotal during DNA repair and during V(D)J recombination as it performs the final DNA-break sealing step. This study explored whether monoallelic LIG4 missense mutations may underlie immunodeficiency and autoimmunity with autosomal dominant inheritance. Extensive flow-cytometric immune-phenotyping was performed. Rare variants of immune system genes were analyzed by whole exome sequencing. DNA repair functionality and T-cell-intrinsic DNA damage tolerance was tested with an ensemble of invitro and in silico tools. Antigen-receptor diversity and autoimmune features were characterized by high-throughput sequencing and autoantibody arrays. Reconstitution of wild-type versus mutant LIG4 were performed in LIG4 knockout Jurkat T cells, and DNA damage tolerance was subsequently assessed. A novel heterozygous LIG4 loss-of-function mutation (p.R580Q), associated with a dominantly inherited familial immune-dysregulation consisting of autoimmune cytopenias, and in the index patient with lymphoproliferation, agammaglobulinemia, and adaptive immune cell infiltration into nonlymphoid organs. Immunophenotyping revealed reduced naive CD4+ T cells and low TCR-Vα7.2+ T cells, while T-/B-cell receptor repertoires showed only mild alterations. Cohort screening identified 2 other nonrelated patients with the monoallelic LIG4 mutation p.A842D recapitulating clinical and immune-phenotypic dysregulations observed in the index family and displaying T-cell-intrinsic DNA damage intolerance. Reconstitution experiments and molecular dynamics simulations categorize both missense mutations as loss-of-function and haploinsufficient. This study provides evidence that certain monoallelic LIG4 mutations may cause human immune dysregulation via haploinsufficiency.

Habitual physical activity in patients born with oesophageal atresia: a multicenter cross-sectional study and comparison to a healthy reference cohort matched for gender and age

Oesophageal atresia (EA) is associated with life-long gastrointestinal and respiratory morbidity and other associated malformations. The aim of this study is to compare physical activity (PA) levels of children and adolescents with and without EA. A validated questionnaire (MoMo-PAQ) was used to evaluate PA in EA patients EA (4–17 years), who were randomly matched for gender and age (1:5) with a representative sample of the Motorik-Modul Longitudinal Study (n = 6233). Sports activity per week (sports index) and minutes of moderate to vigorous physical activity per week (MVPA minutes) were calculated. Correlations between PA and medical factors were analysed. In total, 104 patients and 520 controls were included. Children with EA were significantly less active at higher intensities (mean MPVA minutes 462; 95% confidence interval (CI): 370–554) compared to controls (626; 95% CI: 576–676), although there was no statistically significant difference in the sports index (187; 95% CI: 156–220 versus 220; 95% CI: 203–237). A lower mean weight-for-age and height-for-age, additional urogenital (r = − 0.20, p = 0.04) or anorectal malformation (r = − 0.24, p = 0.01) were associated with fewer MVPA minutes. For other medical factors (prematurity, type of repair, congenital heart disease, skeletal malformation or symptom load), no statistically significant association with PA was found. Conclusion: EA patients participated in PA at a similar level but lower intensities compared to the reference cohort. PA in EA patients was largely independent of medical factors.Trial registration: German Clinical Trials Register (ID: DRKS00025276) on September 6, 2021.What is Known:• Oesophageal atresia is associated with a low body weight and height, delayed development of motor skills and impaired lung function and exercise capacity.What is New:• Patients with oesophageal atresia have a similar amount of sports activity per week but engage significantly less in moderate to vigorous physical activities compared to peers.• Physical activity was associated with weight-for-age and height-for-age, but largely independent of symptom load and other medical factors.

Triazole fungicides induce adipogenesis and repress osteoblastogenesis in zebrafish.

Triazoles are a major group of azole fungicides commonly used in agriculture, veterinary, and human medicine. Maternal exposure to certain triazole anti-fungal medication causes congenital malformations, including skeletal malformations. We hypothesized that triazoles used as pesticides in agriculture also pose a risk of causing skeletal malformations in developing embryos. In this study, teratogenic effects of three commonly used triazoles, cyproconazole, paclobutrazol, and triadimenol, were investigated in zebrafish, Danio rerio. Exposure to the triazole fungicides caused bone and cartilage malformations in developing zebrafish larvae. Data from whole-embryo transcriptomics with cyproconazole suggested that exposure to this compound induces adipogenesis while repressing skeletal development. Confirming this finding, the expression of selected bone and cartilage marker genes were significantly downregulated with triazoles exposure as determined by quantitative PCR. The expression of selected adipogenic genes was upregulated by the triazoles. Furthermore, exposure to each of the three triazoles induced adipogenesis and lipid accumulation in vitro in 3T3-L1 pre-adipocyte cells. In vivo in zebrafish larvae, cyproconazole exposure caused lipid accumulation. These results suggest that exposure to triazoles promotes adipogenesis at the expense of skeletal development, and thus they expand the chemical group of bona fide bone to fat switchers.

Clinical characteristics and offspring developmental malformation of 126 patients with septate uterus.

Clinical characteristics and offspring developmental malformation of 126 patients with septate uterus.

Zoledronic Acid Blocks Overactive Kir6.1/SUR2-Dependent KATP Channels in Skeletal Muscle and Osteoblasts in a Murine Model of Cantú Syndrome.

Cantú syndrome (CS) is caused by the gain of function mutations in the ABCC9 and KCNJ8 genes encoding, respectively, for the sulfonylureas receptor type 2 (SUR2) and the inwardly rectifier potassium channel 6.1 (Kir6.1) of the ATP-sensitive potassium (KATP) channels. CS is a multi-organ condition with a cardiovascular phenotype, neuromuscular symptoms, and skeletal malformations. Glibenclamide has been proposed for use in CS, but even in animals, the drug is incompletely effective against severe mutations, including the Kir6.1wt/V65M. Patch-clamp experiments showed that zoledronic acid (ZOL) fully reduced the whole-cell KATP currents in bone calvaria cells from wild type (WT/WT) and heterozygous Kir6.1wt/V65MCS mice, with IC50 for ZOL block < 1 nM in each case. ZOL fully reduced KATP current in excised patches in skeletal muscle fibers in WT/WT and CS mice, with IC50 of 100 nM in each case. Interestingly, KATP currents in the bone of heterozygous SUR2wt/A478V mice were less sensitive to ZOL inhibition, showing an IC50 of ~500 nM and a slope of ~0.3. In homozygous SUR2A478V/A478V cells, ZOL failed to fully inhibit the KATP currents, causing only ~35% inhibition at 100 μM, but was responsive to glibenclamide. ZOL reduced the KATP currents in Kir6.1wt/VMCS mice in both skeletal muscle and bone cells but was not effective in the SUR2[A478V] mice fibers. These data indicate a subunit specificity of ZOL action that is important for appropriate CS therapies.

The Ethanolic Extract of Piper glabratum Kunth Is Teratogenic and Interferes with the Ossification Process of Swiss Mice Fetuses

Piper glabratum Kunth is a plant traditionally used to treat pain and inflammation in the Mato Grosso do Sul, Brazil. Even pregnant women consume this plant. Toxicology studies of the ethanolic extract from the leaves of P. glabratum (EEPg) could establish the safety of popular use of P. glabratrum. Thus, the effects of the ethanolic extract of leaves of P. glabratum (EEPg) on the reproductive performance and embryofetal development of Swiss mice were evaluated. Pregnant female mice were treated with 100, 1000 and 2000 mg/kg throughout the gestational period by gavage (p.o). The control group received the EEPg vehicle (Tween 80-1%) in the proportion of 0.1 mL/10 g (p.o.). The results demonstrated that EEPg has low maternal toxic potential and does not alter the reproductive performance of females. However, it altered embryofetal development and caused fetal weight reduction (increasing the frequency of small-for-gestational-age fetuses) at the two highest doses. In addition, it interfered with placental weight, placental index and placental efficiency. The frequency of visceral malformations increased by 2.8 times for the lowest dose of EEPg, and skeletal malformations increased by 2.48, 1.89 and 2.11 times for doses of 100, 1000 and 2000 mg/kg of EEPg, respectively. It is noteworthy that 100% of the offspring treated with EEPg showed changes in the ossification process. Thus, it is considered that the EEPg has low maternal toxic potential; it does not alter the reproductive performance of females. However, it is teratogenic and interferes, mainly, in the ossification process, and therefore its use is contraindicated in the gestational period.

16p13.11 microduplication including NDE1 gene in autism spectrum disorder: A case report

IntroductionAutism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by the deficit in communication and social interaction as well as restricted and repetitive interests and behaviors. In addition to the involvement of the environmental component in ASD, it is currently established the significant contribution of genetic factors such us the 16p13.11 duplication. We report the case of a patient carrying this anomaly in whom an ASD was diagnosed, the genetic study of the parents objectified the same anomaly in a clinically healthy father.ObjectivesThis case report aims to expand the clinical findings associated to this genomic abnormality and provide further knowledge of the pathogenic involvement of this duplication.MethodsOur patient, aged 4 years and 10 months, presented developmental peculiarities from birth. The difficulties became more evident with the language delay, the deficit of social interactions and the appearance of motor stereotypies as well as sensory specificities. The diagnosis of ASD was confirmed by the passation of the ADI and the ADOS.Further genetic exploration with a CGH-Array request was performed due to a normal 46 XY karyotype. She objectified a 16p13.11 duplication comprising 25 genes including NDE1. The assessment was completed by the search for potentially associated malformations, particularly cardiac and skeletal. We continued the family investigation with genetic samples for the parents and the siblings finding the same anomaly in the father who does not present any particular phenotype.ResultsThe 16p13.11 duplication is associated with a variable clinical spectrum including behavioral disorders, attention deficit/ hyperactivity disorder, intellectual disability, cardiac and skeletal malformation, epilepsy as well as ASD and language delay presented by our patient. Eight annotated genes are present in this region including NDE1, the candidate gene for the neurocognitive phenotype. This microduplication can be found in the normal population, but it is increasingly detected in patients with ASD, schizophrenia or presenting cognitive disorders due to incomplete penetrance that can explain the presence of the duplication despite the absence of any disorders in the patient’s father.ConclusionsThe link between the 16p13.11 duplication and ASD is increasingly recognized in the literature. The heterogeneity of its clinical expression and especially its incomplete penetrating make genetic counseling difficult. Collaboration between child psychiatrists and geneticists remains essential to detect, link their clinical evaluation and optimize care.Disclosure of InterestNone Declared

Simulated gastric hydrolysis and developmental toxicity of dimethyltin bis(2-ethylhexylthioglycolate) in rats.

Dimethyltin dichloride is used as the putative toxophore for dimethyltin bis-alkylthio esters in a read-across approach. Recent chemical and toxicological investigations challenges this read across as data on dioctyltin bis(2-ethylhexyl thioglycolate) and dibutyltin bis(2-ethylhexyl thioglycolate) showed the dialkyltin thioglycolates do not generate dialkyltin dichloride. Results obtained by 119Sn-NMR spectroscopy demonstrated that dimethyltin bis(2-ethylhexyl thioglycolate), the smallest commercially manufactured dialkyltin thioester molecule of this kind, hydrolyzed to dimethyltin chloro-(2-ethylhexyl) thioglycolate under simulated gastric conditions. These studies did not detect dimethyltin dichloride. Dimethyltin bis(2-ethylhexyl thioglycolate) was administered orally to timed-pregnant Wistar-Han rats in an Arachis oil vehicle at 5, 10, and 25mg/kg/day [Gestation Day 6 (GD6) through GD20] with no maternal deaths observed. At 25mg/kg/day treatment statistically significant reductions occurred in feed consumption (-9%), maternal body weight (-2.4%) and adjusted maternal weight gain (-68%). There were no adverse gestational findings. Maternal thymus weight was significantly reduced in rats at 25mg/kg in the absence of changes in hormone levels of T3, T4 or TSH. There were no effects on fetal growth, no dose-dependent pattern of external, visceral, or skeletal malformations and no toxicologically relevant increase in anatomical variations at any dose group. Based on the obtained experimental data it is concluded that dimethyltin bis(2-ethylhexyl thioglycolate) forms dimethyltin chloro-(2-ethylhexyl thioglycolate), not dimethyltin dichloride, in the stomach environment at pH 1.2, and dimethyltin bis(2-ethylhexyl thioglycolate) was not teratogenic nor fetotoxic in rats. The maternal NOAEL was 10mg/kg/day, and the developmental NOAEL was 25mg/kg/day, the high dose. The maternal LOAEL was 25mg/kg/day based on decreased food consumption, lower adjusted mean body weight gain and reduced maternal thymus weight.

Elucidating the Effects of the Lipids Regulators Fibrates and Statins on the Health Status of Finfish Species: A Review.

The most documented fibrates are gemfibrozil, clofibrate and bezafibrate, while for statins, the majority of the published literature focuses on atorvastatin and simvastatin. The present work reviews previously published research concerning the effects of these hypocholesterolaemic pharmaceuticals on fish, with a particular focus on commercially important species, commonly produced by the European aquaculture industry, specifically in recirculated aquaculture systems (RAS). Overall, results suggest that both acute and chronic exposures to lipid-lowering compounds may have adverse effects on fish, disrupting their capacity to excrete exogenous substances, as well as both lipid metabolism and homeostasis, causing severe ontogenetic and endocrinological abnormalities, leading to hampered reproductive success (e.g., gametogenesis, fecundity), and skeletal or muscular malformations, having serious repercussions on fish health and welfare. Nonetheless, the available literature focusing on the effects of statins or fibrates on commonly farmed fish is still limited, and further research is required to understand the implications of this matter on aquaculture production, global food security and, ultimately, human health.

Adult patient diagnosed with NADSYN1 associated congenital NAD deficiency and analysis of NAD levels to be published in: European Journal of Medical Genetics

IntroductionNicotinamide adenine dinucleotide (NAD) is an essential cosubstrate/coenzyme in multiple cellular redox processes and a substrate in several non-redox reactions. NADSYN1 encodes NAD synthetase 1, an enzyme in the NAD de novo synthesis pathway and the Preiss-Handler pathway, and biallelic pathogenic variants causes NAD deficiency associated with vertebral, cardiac, renal and limb defects. Szot et al. and Kortbawi et al. have reported a total of seven patients with NADSYN1 associated congenital NAD deficiency disorder with the oldest patient being seven years old. Patient dataWe present a male patient age 30 with a height of 130 cm and numerous skeletal malformations including segmentation defects of the spine, rib anomalies and unequal leg length as well as bilateral ptosis, cleft palate and asymmetric dysmorphic facial features. The patient underwent surgery for an aortic stenosis due to a bicuspid valve. No malformations of the kidneys or urinary tract were identified. ResultsTrio exome sequencing revealed a homozygous missense variant in NADSYN1 c.1717G > A (p.Ala573Thr). Both parents were unaffected carriers of the variant. Analysis of NAD levels showed that the patient had a lower NAD pool compared to his unaffected siblings. The NAD pool rose approximately 25% after supplementation with nicotinamide, a NAD precursor for the salvage pathway. ConclusionThe variant was previously reported in four patients and functional analyses by Szot et al. support the pathogenicity of the variant. We report an adult patient with NADSYN1 associated congenital NAD deficiency disorder and expand the phenotypic spectrum. We also present analysis of the NAD levels before and after supplementation with nicotinamide.

Modulatory effect of ginger on skeletal malformations, cell cycle, apoptosis and structural changes in the liver of rat fetuses prenatally exposed to labetalol

BackgroundDrug-induced liver damage with clinical symptoms has been related to labetalol in a number of instances. In addition to having a wide range of anti-inflammatory and antioxidant qualities, ginger also includes biotrace that are crucial in the fight against disease and skeletal deformity. In this study, we hypothesized that prenatal supplementation of ginger (200 mg/kg) attenuates skeletal malformation and hepatotoxicity mediated by labetalol during the organogenesis period. The tested dams were divided into four groups: control, ginger (200 mg/kg), labetalol (300 mg/kg) and combined group (labetalol and ginger at the same doses).ResultsThe labetalol group showed various skeletal abnormalities represented by mandibular hypoplasia, costal separation and retardation in the ossification. Histological and ultrastructural examination of the fetal liver tissue revealed multiple pathological changes. DNA damage, G0/G1 cell cycle arrest and a high percentage of apoptosis were also detected in the fetal hepatocytes from labetalol groups through gel electrophoresis and flow cytometry using PI and annexin V/PI methods, respectively. Administration of ginger after labetalol caused an evident decrease in these skeletal malformations, structural changes, DNA damage, apoptosis and G0/G1 cell cycle arrest.ConclusionsIt can be concluded that ginger has great potential in attenuating the skeletal malformation, structural changes and cyto-genotoxicity of fetal hepatocytes upon prenatal exposure to labetalol.

A novel single-base deletion of the RUNX Family Transcription Factor 2 gene associated with cleidocranial dysplasia.

Cleidocranial dysplasia (CCD) is a rare, autosomal dominant hereditary disorder characterized by skeletal malformations and dental abnormalities. The purpose of this study was to explore the functional role of a novel mutation in the pathogenesis of CCD. Genomic DNA was extracted from peripheral blood mononuclear cells collected from family members of a Chinese patient with CCD. An analysis of their RUNX Family Transcription Factor 2 (RUNX2) gene sequences was performed by PCR amplification and Sanger sequencing. The function of the mutant RUNX2 was studied by bioinformatics, real-time PCR, western blotting, and subcellular localization analysis. Sanger sequencing identified a novel single-base deletion (NM_001024630.4:c.132delG;NP_001019801.3: Val45Trpfs* 99) in the RUNX2 gene present in the Chinese patient with CCD. In vitro, functional studies showed altered protein localization and increased expression of mutant RUNX2 mRNA and mutant Runt-related transcription factor 2 (RUNX2). Luciferase reporter assay demonstrated that the novel RUNX2 mutations significantly increased the transactivation activity of RUNX2 on the osteocalcin gene promoter. In conclusion, we identified a patient with sporadic CCD carrying a novel deletion/frameshift mutation of the RUNX2 gene and performed screening and functional analyses to determine the cause of the CCD phenotype. This study provides new insights into the pathogenesis of CCD.3.

Prenatal developmental toxicity of gardenia blue, a natural food colorant, in rats and rabbits

Introduction Gardenia blue is a colorant widely used in Asia in food and beverages. The objectives of the present studies were to evaluate the maternal and prenatal embryo-fetal developmental toxicity of gardenia blue in rats and rabbits. Methods Sprague Dawley rats and New Zealand White rabbits were administered gardenia blue daily by oral gavage at doses of 0 (deionized water vehicle), 500, 1000 or 2000 mg/kg/day on Gestation Days 6 through 20 (rats) and 7 through 28 (rabbits). Endpoints evaluated included clinical observations, body weight, food consumption, thyroid hormones (rats), thyroid weights and histopathology (rats), gross pathologic changes, ovarian and uterine observations, fetal weight and anogenital distance (rats) and fetal morphology (external, visceral and skeletal). Results Treatment related maternal findings attributed to the blue/dark color of the test substance included body surface staining, and dark/blue discoloration of the kidneys, gastrointestinal track and mesenteric lymph nodes at all or most doses in the rat and/or rabbit. Slight reductions in food consumption without effects on body weight were also observed in rats at all doses and in rabbits at 2000 mg/kg/day. There were no treatment related effects on maintenance of pregnancy, postimplantation loss, litter size, fetal weight and anogenital distance, or fetal external, visceral, or skeletal malformations and variations. Conclusions Based on these results, the maternal and developmental no-observed-adverse-effect level for gardenia blue in rats and rabbits was ≥2000 mg/kg/day.

Hypomorphic and dominant-negative impact of truncated SOX9 dysregulates Hedgehog–Wnt signaling, causing campomelia

Haploinsufficiency for SOX9, the master chondrogenesis transcription factor, can underlie campomelic dysplasia (CD), an autosomal dominant skeletal malformation syndrome, because heterozygous Sox9 null mice recapitulate the bent limb (campomelia) and some other phenotypes associated with CD. However, invitro cell assays suggest haploinsufficiency may not apply for certain mutations, notably those that truncate the protein, but in these cases invivo evidence is lacking and underlying mechanisms are unknown. Here, using conditional mouse mutants, we compared the impact of a heterozygous Sox9 null mutation (Sox9+/-) with the Sox9+/Y440X CD mutation that truncates the C-terminal transactivation domain but spares the DNA-binding domain. While some Sox9+/Y440X mice survived, all Sox9+/- mice died perinatally. However, the skeletal defects were more severe and IHH signaling in developing limb cartilage was significantly enhanced in Sox9+/Y440X compared with Sox9+/-. Activating Sox9Y440X specifically in the chondrocyte-osteoblast lineage caused milder campomelia, and revealed cell- and noncell autonomous mechanisms acting on chondrocyte differentiation and osteogenesis in the perichondrium. Transcriptome analyses of developing Sox9+/Y440X limbs revealed dysregulated expression of genes for the extracellular matrix, as well as changes consistent with aberrant WNT and HH signaling. SOX9Y440X failed to interact with β-catenin and was unable to suppress transactivation of Ihh in cell-based assays. We propose enhanced HH signaling in the adjacent perichondrium induces asymmetrically localized excessive perichondrial osteogenesis resulting in campomelia. Our study implicates combined haploinsufficiency/hypomorphic and dominant-negative actions of SOX9Y440X, cell-autonomous and noncell autonomous mechanisms, and dysregulated WNT and HH signaling, as the cause of human campomelia.

Infertility treatment for Chinese women with P450 oxidoreductase deficiency: Prospect on clinical management from IVF to FET.

Cytochrome P450 oxidoreductase deficiency (PORD) is a rare recessive disease with multiple clinical manifestations, which is usually diagnosed in neonates and children because of ambiguous genitalia or skeletal malformations. Moreover, the paucity of studies does not allow us to establish whether adult-onset PORD is associated with infertility. Here, we report clinical and laboratory findings in two phenotypically normal women diagnosed with PORD who underwent in vitro fertilization (IVF) and frozen embryo transfer (FET). We modified the gonadotropin stimulation protocol during controlled ovarian hyperstimulation (COH) and suggest the use of the vaginal 17β-estradiol route for endometrium preparation in hormone replacement therapy (HRT) cycles. We presume that PORD may be associated with infertility in several aspects, including disordered steroidogenesis, endometrium impairment, attenuation of drug metabolism, and the high risk of miscarriage. Our observations will help the early diagnosis and make a tailored approach to infertility management in adult-onset PORD.

Teratogenic Toxicity of Ethanol Extract of Mahkota Dewa Fruit Flesh (Phaleria macrocarpa (Scheff.) Boerl)

Mahkota Dewa is known as one of the medicinal plants in Indonesia. It contains bioactive compounds and potentially has pharmacological activity. This study aimed to determine the possible toxic effects of Mahkota DewaThe ethanol extract of Mahkota Dewa fruit flesh was given to rats. Group I (control 0.5% Na-CMC), groups II, III and IV as a treatment group (Mahkota Dewa at doses of 100, 500 and 1000 mg/kg bw, respectively). Group V as a control satellite (Na-CMC 0, 5%) and group VI as a satellite of the ethanol extract at a dose of 1000 mg/kg bw., Each groups were treated on day 6 to 15 of gestational. On the 19th day getation, the rats were dissected and observed the skeletal malformations. The data were analyzed using one-way ANOVA followed by a post-hoc tukey. The results toxicity showed that no fetuses had external malformations. On fetal skeletal appearance, all skeletal bone preparations were normal. Based on the study, the ethanolic extract of Mahkota Dewa Fruit Flesh did not cause a teratogenic effect on the fetus during the organogenesis period at doses of 100 mg/kg bw, 500 mg/kg bw, and 1000 mg/kg bw

Role of transcerebellar diameter in fetal growth assessment and its correlation with conventional biometry-An institutional study

Background: Accurate assessment of gestational age (GA) for fetal development is of paramount importance in management of pregnancy and for favorable perinatal outcome. The estimation of GA by fetal biometry using Biparietal Diameter (BPD), Head Circumference (HC), Abdominal Circumference (AC) and Femoral Length (FL) in estimation of gestational age (GA) is routinely followed. However, these parameters have limitations. BPD and HC are not reliable in third trimester; FL may be reduced in IUGR and skeletal malformations. Thus we evaluated the Transcerebellar diameter (TCD) as an alternate indicator for fetal growth and GA estimation. Aim: To validate the accuracy of TCD with fetal biometry and calculated ultrasound age (CUA) in assessment of fetal growth and GA in between 18 to 40 weeks of pregnancy. Materials and Methods: The prospective study was carried out with 200 pregnant women between 18 to 40 weeks referred to Radiology department ((JJM Medical College), for antenatal scanning. The subjects were divided into two groups (second and third trimesters). TCD was measured along with other routine parameter for growth assessment. Thus, calculated gestational age (GA) using TCD was compared with GA by Last menstrual period (LMP) and calculated ultrasound age (CUA).