Nonlethal Malformations Research Articles

Implementation and Assessment of a Laparotomy-Assisted Three-Port Fetoscopic Spina Bifida Repair Program.

Open spina bifida (OSB) is a congenital, non-lethal malformation with multifactorial etiology. Fetal therapy can be offered under certain conditions to parents after accurate prenatal diagnostic and interdisciplinary counseling. Since the advent of prenatal OSB surgery, various modifications of the original surgical techniques have evolved, including laparotomy-assisted fetoscopic repair. After a two-year preparation time, the team at the University of Giessen and Marburg (UKGM) became the first center to provide a three-port, three-layer fetoscopic repair of OSB via a laparotomy-assisted approach in the German-speaking area. We point out that under the guidance of experienced centers and by intensive multidisciplinary preparation and training, a previously described and applied technique could be transferred to a different setting.

Embryonic exposure to butylparaben and propylparaben induced developmental toxicity and triggered anxiety‐like neurobehavioral response associated with oxidative stress and apoptosis in the head of zebrafish larvae

Parabens are synthetic antimicrobial compounds used as a preservative for extending the shelf life of food, pharmaceutical and cosmetic products. The alkyl chain length of the paraben esters positively correlates with their antimicrobial property. Hence, long-chain paraben esters, namely butylparaben and propylparaben, are used in combination as they have better solubility and antimicrobial efficacy. Extensive use of parabens has now resulted in the ubiquitous presence of these compounds in various human and environmental matrices. During early life, exposure to environmental contaminants is known to cause oxidative-stress mediated apoptosis in developing organs. The brain being one of the high oxygen-consuming, metabolically active and lipid-rich organ, it is primarily susceptible to reactive oxygen species (ROS) and lipid peroxidation (LP) induced neuronal cell death. The primary cause for the impairment in cognitive and emotional neurobehvioural outcomes in neurodegenerative disease was found to be associated with neuronal apoptosis. The present study aimed to study butylparaben and propylparaben's effect on zebrafish during early embryonic stages. Besides this, the association between alteration in anxiety-like neurobehavioral response with oxidative stress and antioxidant status in head region was also studied. The study results showed variation in the toxic signature left by butylparaben and propylparaben on developmental parameters such as hatching rate, survival and non-lethal malformations in a time-dependent manner. Data from the light-dark preference test showed embryonic exposure to butylparaben and propylparaben to trigger anxiety-like behavior in zebrafish larvae. In addition, a significant increase in intracellular ROS and LP levels correlated with suppressed antioxidant enzymes: superoxide dismutases (SOD), catalases (CAT), Glutathione peroxidase (GPx), glutathione S-transferase (GST), and Glutathione (GSH) activity in the head region of the zebrafish larvae. Acetylcholinesterase (AChE) activity was also suppressed in the exposed groups, along with increased nitric oxide production. The overall observations show increased oxidative stress indices correlating with upregulated expression of apoptotic cells in a dose-dependent manner. Collectively, our findings reveal butylparaben and propylparaben as an anxiogenic neuroactive compound capable of inducing anxiety-like behavior through a mechanism involving oxidative-stress-induced apoptosis in the head of zebrafish larvae, which suggests a potential hazard to the early life of zebrafish and this can be extrapolated to human health as well.

Acute toxicity of polybrominated diphenyl ethers (PBDEs) for turbot (Psetta maxima) early life stages (ELS)

The environmental presence of polybrominated diphenyl ethers (PBDEs), among which BDE-47 and BDE-99 are particularly abundant, makes toxicity data necessary to assess the hazard risk posed by PBDE to aquatic organisms. This study examines the effects of BDE-47 and BDE-99 on embryo-larval stages of the marine flatfish turbot. The turbot embryos were exposed at nominal concentrations of BDE-47 and BDE-99 for 6 days. Selected dose levels were relevant for investigating sublethal and lethal effects. Both tested compounds caused lethal toxicity as well as non-lethal malformations during embryo development. We found a high toxic potency of BDE-47 compared to BDE-99 (LC₅₀ values for embryos and larvae, respectively, BDE-47: 27.35 and 14.13 μg L⁻¹; BDE-99: 38.28 and 29.64 μg L⁻¹). The present study shows high sensitivity of fish early life stages (ELS) to PBDE compounds. Based on environmental concentrations of dissolved PBDEs from various aquatic ecosystems, waterborne BDE-47 and BDE-99 pose little risk of acute toxicity to marine fish at relevant environmental concentrations. Turbot fish ELS proved to be an excellent model for the study of ecotoxicity of contaminants in seawater. The results demonstrate harmful effects of PBDE on turbot ELS at concentrations in the range of parts per billion units. In the perspective of risk assessment, ELS endpoints provide rapid, cost-effective and ecologically relevant information, and links should be sought between these short-term tests and effects of long-term exposures in more realistic scenarios.

In utero surgery – current state of the art – part II

SummaryBackgroundFetal surgery, also referred to as in utero, prenatal or intrauterine surgery, consists of treatment of congenital malformations during the fetal period. The idea of treating malformations diagnosed in the course of intrauterine life dates back to 1963, when Lilly performed the first blood transfusion in a fetus. Since then it has been introduced as a treatment option in a series of lethal malformations. Efforts are being made to treat nonlethal malformations by means of fetal surgery.Material/MethodsA comprehensive search of the literature using MEDLINE and PubMed between 1925 and February 2009 was performed. Search terms for MEDLINE and PubMed were: fetal surgery, foetal surgery, in utero surgery, prenatal surgery, and in utero treatment. In addition, information was obtained at Web sites of the International Medicine and Surgery Society and the University of California Fetal Treatment Centre.ResultsAuthors’ attention focused on the survey of indications to intrauterine operations. We outline potential directions of its development, quoting the groundwork of the most experienced researchers and clinicians. Moreover, owing to the authors’ interest in plastic surgery, some remarks on the role of intrauterine medicine and surgery in this branch of medicine are made.ConclusionsIn utero surgery may be regarded as an efficient tool of preventive medicine. It offers some advantages that no other branch of medicine may offer. However, its implementation is more troublesome than in the past, therefore only selected cases may benefit from its advantages. Nevertheless, current tendencies are to include, after thorough evaluation of benefit-risk ratio, some new indications for fetal surgery.

Lethal and sublethal effects of polybrominated diphenyl ethers (PBDEs) for turbot (Psetta maxima) early life stage (ELS)

A new toxic menace, polybrominated diphenyl ethers (PBDEs), is being detected in the aquatic environment all over the world. The environmental presence of PBDEs and its entry into the environment as BDE-47 and -99 make quality aquatic toxicity data necessary to assess the aquatic hazard risk of PBDs. This study examines the effects of three PBDE-47 and -99 on embryo and larval stages of the marine flatfish turbot (Psetta maxima). The acute toxicity of the three PBDEs was examined and NOEC, LOEC, LC10 and LC50 were calculated. All tested compounds caused lethal as well as nonlethal malformations during embryo development. The effects of PBDEs in the different life stages of turbot were analysed. PBDEs seemed to be teratogenic at concentrations higher than 8.14 and 16.12 µgL-1 for BDE-47 and -99 respectively, leading to prolonged delays in embryo development and consequent death (at 48 h), as well as severe malformations and mortality of larvae. PBDEs showed a higher acute toxicity for embryo-larvae (LC50 for lethal endpoints to embryos 27.35 µg BDE-47 L-1 and 38.28 µg BDE-99 L-1, and 14.13 and 29.64 µg L-1 for BDE-47 and -99, respectively for larvae). Generally, the BDE-47 seemed to cause adverse effects at comparatively low dose rates, whereas much higher doses were needed to cause the same effects with BDE-99. The results of the present study show that the acute toxicity of PBDEs decreases as the degree of bromination increases, since the order of toxicity was BDE-47 BDE-99. The major isomers also exhibited a clear toxic potential for turbot ELS although at high concentrations, above solubility saturation, pointing at particulate matter as the main via of uptake for those hydrophobic molecules.

Lethal and sublethal effects of polybrominated diphenyl ethers (PBDEs) for turbot (Psetta maxima) early life stage (ELS)

AbstractA new toxic menace, polybrominated diphenyl ethers (PBDEs), is being detected in the aquatic environment all over the world. The environmental presence of PBDEs and its entry into the environment as BDE-47 and -99 make quality aquatic toxicity data necessary to assess the aquatic hazard risk of PBDs. This study examines the effects of three PBDE-47 and -99 on embryo and larval stages of the marine flatfish turbot (Psetta maxima). The acute toxicity of the three PBDEs was examined and NOEC, LOEC, LC10 and LC50 were calculated. All tested compounds caused lethal as well as nonlethal malformations during embryo development. The effects of PBDEs in the different life stages of turbot were analysed. PBDEs seemed to be teratogenic at concentrations higher than 8.14 and 16.12 µgL-1 for BDE-47 and -99 respectively, leading to prolonged delays in embryo development and consequent death (at 48 h), as well as severe malformations and mortality of larvae. PBDEs showed a higher acute toxicity for embryo-larvae (LC50 for lethal endpoints to embryos 27.35 µg BDE-47 L-1 and 38.28 µg BDE-99 L-1, and 14.13 and 29.64 µg L-1 for BDE-47 and -99, respectively for larvae). Generally, the BDE-47 seemed to cause adverse effects at comparatively low dose rates, whereas much higher doses were needed to cause the same effects with BDE-99. The results of the present study show that the acute toxicity of PBDEs decreases as the degree of bromination increases, since the order of toxicity was BDE-47 BDE-99. The major isomers also exhibited a clear toxic potential for turbot ELS although at high concentrations, above solubility saturation, pointing at particulate matter as the main via of uptake for those hydrophobic molecules.

Lethal and sublethal effects of polybrominated diphenyl ethers (PBDEs) for turbot (Psetta maxima) early life stage (ELS)

A new toxic menace, polybrominated diphenyl ethers (PBDEs), is being detected in the aquatic environment all over the world. The environmental presence of PBDEs and its entry into the environment as BDE-47 and -99 make quality aquatic toxicity data necessary to assess the aquatic hazard risk of PBDs. This study examines the effects of three PBDE-47 and -99 on embryo and larval stages of the marine flatfish turbot (Psetta maxima). The acute toxicity of the th ree PBDEs was examined and NOEC, LOEC, LC10 and LC50 were calculated. All tested compounds caused lethal as well as nonlethal malformations during embryo development. The effects of PBDEs in the different life stages of turbot were analysed. PBDEs seemed to be teratogenic at concentrations higher than 8.14 and 16.12 µgL -1 for BDE-47 and -99 respectively, leading to prolonged delays in embryo development and consequent death (at 48 h), as well as severe malformations and mortality of larvae. PBDEs showed a higher acute toxicity for embryo-larvae (LC50 for lethal endpoints to embryos 2 7.35 µg BDE-47 L -1 and 38.28 µg BDE-99 L -1 , and 14.13 and 29.64 µg L -1 for BDE-47 and -99, respectively for larvae). Generally, the BDE-47 seemed to cause adverse effects at comparatively low dose rates, whereas much higher doses were needed to cause the same effects with BDE-99. The results of the present study show that the acute toxicity of PBDEs decreases as the degree of bromination incre ases, since the order of toxi city was BDE47˃ BDE-99. The major isomers also exhibited a clear toxic potential for turbot ELS although at high concentrations, above solubility saturation, pointing at particulate matter as the main via of uptake fo r those hydrophobic molecules.

Developmental toxicity of bifenthrin in embryo-larval stages of zebrafish

Bifenthrin (BF) is widely used as a miticide in orchards, nurseries and homes due to its great photostability and insecticidal activity. Recently, extensive research has been conducted on the toxicity of BF in in vitro and in vivo assays. However, no data is so far available regarding the developmental toxicity of BF to fish in early life stages. In this study, the developmental effects of BF were evaluated in embryo-larval zebrafish. At specified stages (24, 48, 72, and 96 hpf), spontaneous movement, survival and hatching as well as non-lethal malformation like curved body axis or edema were described in detail. No significant lethal effects of the treatment group compared to the control occurred except for the highest concentration group exposed for 96 h. The hatching process was accelerated by BF in a concentration-dependent way, correlated with increasing spontaneous movement. Developmental abnormalities were observed for the test compound with 96-h EC 50 of 256 μg L −1 for pericardial edema, and 109 μg L −1 for curved body axis. Results from locomotor assays showed that zebrafish larvae of 96 hpf exhibited impaired swimming behaviour after exposure to 50, 100, and 200 μg L −1 from 3 to 84 hpf. After being cultured in the BF-free embryo medium for one more day till 120 hpf, larvae from the 50 μg L −1group seemed to have recovered and showed no difference in the swimming behaviour compared to the control while animals of the two higher concentration groups were exhausted and swam in significantly lower speed. Furthermore, reverse transcription real-time PCR results showed that vitellogenin I expressions were significantly induced in larval zebrafish exposed to 150 μg L −1 BF for 72 h, indicating the disruption of the endocrine level. In summary, our studies showed that BF was developmentally toxic to zebrafish in early life stage after short-term exposure to sublethal concentrations and had the ability to impair the individual behaviours which are of great importance in the assessment of their ecological fitness.

Acute toxicity of 353-nonylphenol and its metabolites for zebrafish embryos

Nonylphenol (NP) can be detected in the aquatic environment all over the world. It is applied as a technical mixture of isomers of which 353-NP is the most relevant both in terms of abundance (about 20% of total mass) and endocrine potential. 353-NP is metabolised in sewage sludge. The aims of the present study were to determine and to compare the acute toxicity of t-NP, 353-NP and its metabolites as well as to discuss if the toxicity of 353-NP changes during degradation. 353-NP and two of its metabolites were synthesised. The zebrafish embryo test was performed according to standard protocols. Several lethal and non-lethal endpoints during embryonal development were reported. NOEL, LOEL and EC50 were calculated. All tested compounds caused lethal as well as non-lethal malformations during embryo development. 353-NP showed a higher toxicity (EC50 for lethal endpoints 6.7 mg/L) compared to its metabolites 4-(3.5-dimethyl-3-heptyl)-2-nitrophenol (EC50 13.3 mg/L) and 4-(3,5-dimethyl-3-heptyl)-2-bromophenol (EC50 27.1 mg/L). In surface water, concentrations of NP are far below the NOEC identified by the zebrafish embryo test. However, in soils and sewage sludge, concentrations may reach or even exceed these concentrations. Therefore, sludge-treated sites close to surface waters should be analysed for NP and its metabolites in order to detect an unduly high contamination due to runoff events. The results of the present study point out that the toxicity of 353-NP probably declines during metabolisation in water, sediment and soil, but does not vanish since the major metabolites exhibit a clear toxic potential for zebrafish embryos. Metabolites of environmental pollutants should be included in the ecotoxicological test strategy for a proper risk assessment.

Deformities in larval gilthead sea bream ( Sparus aurata): A qualitative and quantitative analysis using geometric morphometrics

Deformities in commercially raised fish are a common source of downgrading of product value. During the intensive rearing of gilthead sea bream ( Sparus aurata), opercular deformities are the most commonly observed type of deformation (affecting up to 80% of the fisheries stock), sometimes showing a severe inward folding of the operculum. They are non-lethal malformations that appear during the larval stage but affect growth rate and morphology, with a significant economic loss as a consequence. In order to exploratory quantify and qualify these deformities, geometric morphometric analyses were performed on the external morphology from larvae with an age ranging from 50 to 69 days post-hatching (DPH), and on the cranial skeleton of 110 DPH old juveniles. The results showed several osteological cranial shifts and a striking left–right independency associated with deoperculation. Even though a significant size difference was observed at 65 DPH between normal and deoperculated specimens, allometries during the examined growth stages still appear to be very similar in normal and deoperculated specimens. At 69 DPH deoperculated specimens differed significantly from the normal specimens in their external morphology based on its shape variables, but the results suggest that discrimination is possible from earlier stages. Further analyses are needed, but the usefulness of this approach towards developing an early detection tool could be demonstrated.

Toxic Effects of Brominated Indoles and Phenols on Zebrafish Embryos

Organobromine compounds in the marine environment have been the focus of growing attention in past years. In contrast to anthropogenic brominated flame retardants, other brominated compounds are produced naturally, e.g., by common polychaete worms and algae. Brominated phenols and indoles assumed to be of biogenic origin have been detected in water and sediment extracts from the German Bight. These substances as well as some of their isomers have been tested with the zebrafish embryo test and were found to cause lethal as well as nonlethal malformations. The zebrafish test was able to detect a log K(OW)-related toxicity for bromophenols, suggesting nonpolar narcosis as a major mode of action. Different effect patterns could be observed for brominated indoles and bromophenols. The comparison of effective concentrations in the zebrafish embryo test with the concentrations determined in water samples suggests the possibility that brominated indoles may affect early life stages of marine fish species in the North Sea.

66 Evaluating Illness Severity for Very Low Birth Weight Infants: Crib or Crib-II ?

Background: The clinical risk index for babies (CRIB) is a risk-adjustment instrument used worldwide to determine illness severity in infants born at less than 31 weeks' gestation, or having 1500 g birth weight or lower. However, the appropriateness of CRIB has been questioned, and CRIB has very recently been updated and simplified into a five-items scoring system (CRIB-II) for infants born at less than 32 weeks' gestation, in order to improve its accuracy. Here, we compared the accuracy of CRIB and CRIB-II scores in predicting mortality for VLBW infants in a small unit. Methods: A total of 147 NICU-admitted VLBW infants with data available for both CRIB and CRIB-II were enrolled in the study (M:70, F:77; mean ± SD: 28.6 ± 2.5 weeks; birth weight: 1070 ± 325 g). Major non-lethal malformations were present in 3 (2.04%) infants and minor birth defects were observed in 7 (4.76%). Extremely low birth weight infants were 59 (40.15% of total; M:27, F:32). Newborns with lethal congenital abnormalities, metabolic disorders or undergoing complex cardiac surgery were excluded. Both CRIB and CRIB-II scores were calculated for each newborn. Neonatal mortality (death before hospital discharge) was selected as the outcome measure. Results: The median CRIB and CRIB-II scores were 2.0 (range:0–16), and 8.0 (range:3–20), respectively. A significant positive correlation between CRIB and CRIB-II was observed (P <0.0001), with a mortality rate of 11.56% (17/147). Newborns with unfavourable outcome (M:8, F:9) had a significantly lower birth weight (696±205 g vs. 1130±300; p<0.0001), gestational age (25.7±2.9 weeks vs. 29.08±2.1 weeks; p<0.0001), CRIB (median [25th-75th percentiles]: 8.0 vs. 2.0, P<0.0001) and CRIB-II (14.0 vs. 8.0, P <0.0001) than surviving infants. Mean AUCs for CRIB, CRIB-II, gestational age and birth weight in identifying neonatal mortality in VLBW infants ranged from 0.924 (CRIB) to 0.869 (gestational age). No significant differences were observed for the AUCs of CRIB vs. CRIB-II (P=0.34), CRIB vs. gestational age (P=0.18), CRIB vs. birthweight (P=0.24), CRIB-II vs. gestational age (P=0.62), or CRIB-II vs. birth weight (P=86). Conclusion: Our main findings suggest that CRIB and CRIB-II have comparable accuracy in predicting neonatal mortality in VLBW infants. Although our observed mortality rate is higher than that reported for the original CRIB-II population, differences between the two proportions are not statistically significant (P =0.16).

Causes of death in neonates born with a heart malformation

All deceased infants were autopsied in a territory with a population of 1 220 000 inhabitants and a mean of 17 410 live births a year. During a period of 25 years (which preceded the introduction of advanced neonatal cardiac surgery), 393 neonates (6% of all deceased neonates) had died in whom a congenital heart malformation was found at autopsy. 118 (30%) of deaths were due to noncardiac causes. Lethal congenital noncardiac malformations caused death in 38 (10%) neonates. A perinatal disease (including birth trauma) was the cause in 65 (16%) neonates. An association of a noncardiac congenital malformation with a perinatal disease was involved in 6 (2%) neonates and a birth weight lower than 1000 g (together with a lethal perinatal disease or alone) was considered responsible in 9 (2%) neonates. Noncardiac but not unambiguously lethal malformations (mostly multiple) could at least have contributed to the fatal outcome in a further 86 (22%) neonates. A low birth weight could have contributed to death in 102 (26%) neonates, particularly in 42 (11%) neonates with a low birth weight associated with multiple noncardiac nonlethal malformations. A most probable or possible noncardiac cause of death was found in all cases of atrial septal defect, in 93% of tetralogies of Fallot, in 84% of ventricular septal defects, in 72% of hearts with double outlet right ventricle and in 57% of those with aortic coarctation. A congenital heart malformation was considered responsible for a fatal outcome in 82% of cases with discordant ventriculoarterial connexions (“transposition of the great arteries”), 79% of hypoplastic left hearts, 68% of pulmonary atresias and 53% of tricuspid atresias. The high proportion of noncardiac death noted to occur in the first postnatal day was found rapidly to decrease in the following days.