Detection of reactive oxygen species (ROS) and apoptosis in human fragmented embryos

In human in-vitro fertilization (IVF)-embryo transfer, the in-vitro culture environment differs from in-vivo conditions in that the oxygen concentration is higher, and in such conditions the mouse embryos show a higher concentration of reactive oxygen species (ROS) in simple culture media. ROS are believed to cause damage to cell membranes and DNA fragmentation in somatic cells. This study was conducted to ascertain the level of H2O2 concentration within embryos and the morphological features of cell damage induced by H2O2. A total of 62 human oocytes and embryos (31 fragmented, 15 non-fragmented embryos, 16 unfertilized oocytes) was obtained from the IVF-embryo transfer programme. The relative intensity of H2O2 concentrations within embryos was measured using 2',7'-dichlorodihydrofluorescein diacetate by Quanti cell 500 fluorescence imaging and DNA fragmentation was observed with transmission electron microscopy and an in-situ apoptosis detection kit. The H2O2 concentrations were significantly higher in fragmented embryos (72.21 +/- 9.62, mean +/- SEM) compared to non-fragmented embryos (31.30 +/- 3.50, P < 0.05) and unfertilized oocytes (30.75 +/- 2.67, P < 0.05). Apoptosis was observed only in fragmented embryos, and was absent in non-fragmented embryos. Electron microscopic findings confirmed apoptotic bodies and cytoplasmic condensation in the fragmented blastomeres. We conclude that there is a direct relationship between increased H2O2 concentration and apoptosis, and that further studies should be undertaken to confirm these findings.      

De novo 16p13.11 microdeletion identified by high-resolution array CGH in a fetus with increased nuchal translucency

Objective  We investigated the application of high-resolution microarray-based comparative genomic hybridisation (array CGH) on a fetus showing increased nuchal translucency (NT).
Design  Case study.
Setting  Tertiary referral obstetrics unit.
Sample  Pregnant woman attended the antenatal clinic.
Methods  Conventional karyotyping and genetic test was carried out for the alpha-globin gene. High-resolution array CGH using the high-density 244K Agilent microarray was performed on fetal blood sample by cordocentesis to investigate the possibility of any genomic imbalance.
Main outcome measures  Detection of chromosomal abnormality.
Results  Karyotyping analysis showed 46,XY. Molecular genetic diagnosis confirms the fetus has Hb-H constant spring disease but cannot explain the increased NT to 3.2 mm. Array CGH analysis discovered a 1.32-Mb microdeletion on chromosome 16p13.11. Deletion at 16p13.11 has been implicated to predispose to autism and/or mental retardation. Baby was delivered at 40 weeks of gestation, and follow up was carried out at 3 months of age without sign of mental retardation/developmental delay.
Conclusions  This case study demonstrated that array CGH can accurately calibrate the size and identify de novo interstitial chromosome imbalances. However, the presence of chromosome copy variants with unknown clinical significance currently limits its wider scale application in prenatal diagnosis and needs further investigations.     

Is homozygous α-thalassaemia a lethal condition in the 1990s?

Two cases of homozygous α-thalassaemia who received active treatment in accordance with parental wishes are reported. One infant survived and the other, although successfully weaned off mechanical respiratory support, unexpectedly developed portal vein thrombosis and died. Homozygous a-thalassaemia, a condition previously considered to be universally fatal, and an indication for therapeutic abortion, is now potentially curable with advances in diagnostic technology and treatment. However, active management of these cases raises serious ethical questions and has major financial implications on the health-care system. Invasive prenatal and intensive postnatal interventions should remain experimental and cannot be recommended as routine clinical practice until the questions of long-term neurodevelopmental outcome, and the morbidity and mortality associated with bone-marrow transplantation have been fully addressed. As a result of advances in information technology, more and more parents of affected foetuses are likely to request active treatment.     

Dietitians' and nutritionists' knowledge and views on aspects of health claims regulation in the UK: Do we inadvertently shoot the messenger?

Article 12(c) of the Nutrition and Health Claims Regulation (NHCR) prohibits authorised health claims in consumer-facing commercial communications which make reference to the recommendations of individual doctors or health professionals. However, this has been controversial amongst dietitians and nutritionists who work in commercial settings. Given the lack of empirical data, a survey was conducted amongst UK-based nutrition professionals to assess their knowledge of, and attitudes to, Article 12(c). The findings revealed confusion about the scope of the regulation and how it applies to working practices, with a considerable proportion of respondents being unable to recognise examples of commercial communications or health claims, indicating a need for additional training. There was also a broad interpretation of what nutrition professionals could, and could not, say about a hypothetical food product. This paper explores current guidance in Great Britain and debates the proportionality and fairness of Article 12(c), which, at present, does not regulate authorised health claims made by influencers or celebrities in commercial communications to consumers. It could be argued that consumers are better protected by the articulation of health claims by nutrition professionals who are guided by codes of practice rather than by unqualified, unregulated individuals. Hence, it is essential to level the regulatory playing field either by revising the NHCR to amend Article 12(c) or by updating the guidance to apply an interpretation of the Article's intention which enables a broader role for nutrition professionals in commercial communications. Such action would also be consistent with the UK's better regulation agenda to ensure evidence-based, proportionate regulation for industry.     

The appearance and distribution of microglia in the developing retina of the rat

We have examined the development of microglia in the rat retina, using a peroxidase-conjugated lectin derived from Griffonia simplicifolia. Retinas were studied from animals aged from E(embryonic day)12, just after the invagination of the optic cup and prior to the closure of the optic fissure, to adulthood. The lectin also proved a sensitive label for the endothelial cells of the developing retina. Our results provide some support for the view that microglia are derived from the monocyte-macrophage series of blood cells. At E12, most labeled cells were found at the vitreal surface, suggesting that they had come from the hyaloid circulation, while some had entered the retina and appeared to be migrating towards its ventricular surface. From E14 to early postnatal ages, most labeled cells had processes and resembled the amoeboid microglial cells described in silver carbonate staining studies (Ling, 1982). The number of labeled cells rose from about 700 to E14 to a peak of about 27,000 at P(postnatal day)7, and fell to about 19,600 by P12. As early as E16, a regularity was apparent in the distribution of microglial cells over the surface of the retina, the cells tending to avoid each other. Microglial cells are found throughout the thickness of the very young retina, but as the layers of the retina differentiate, they are increasingly restricted to the inner half of the retina. Our findings indicate that microglia enter the retina well before the period of neuronal death, making it unlikely that they invade the retina solely in response to cell death. Our results confirm however that, once in the retina, microglia become associated with, and appear to phagocytose, the pyknotic debris which appears during the period of neuronal death. They also become closely associated with the retinal vasculature. In the adult, the intensity of the labeling of microglia was much reduced. Those cells which were labeled appeared more differentiated, resembling the "resting microglia" described in earlier studies.     

Obesity: new insight into the anthropometric classification of fat distribution shown by computed tomography

Twenty eight women presenting for routine computed tomography had their waist, hip, and thigh circumferences measured. The ratio of the area of intra-abdominal fat to the area of subcutaneous fat shown in the computed tomogram taken at the umbilical level was calculated and found to correlate highly significantly with the ratio of waist to hip circumference. The correlation between these two ratios remained significant after allowing for the degree of obesity (weight (kg)/height (m)2) and age. In contrast, there was no significant correlation between the ratio of intra-abdominal to subcutaneous fat and degree of obesity. A high ratio of waist to hip circumference has been shown to be associated with a high proportion of intra-abdominal fat. Thus women with a centralised distribution of fat (high waist to hip ratio: "apples") tend to have a greater proportion of their fat in the intra-abdominal depot than do women with a peripheral fat distribution (low waist to hip ratio: "pears"). The metabolic complications of obesity, which are associated with a high ratio of waist to hip circumference, may therefore relate specifically to the amount of intra-abdominal fat.