Use of preimplantation genetic diagnosis to produce tissue donors: an irreconcilable dichotomy?

In 2002, the Human Fertilisation and Embryology Authority (HFEA) published its decision in the case of Raj and Shahana Hashmi. The couple had sought to use preimplantation genetic diagnosis (PGD) to have a child that could be a viable donor for their son, Zain, who has thalassaemia. The HFEA decided that PGD could, in the present case, go ahead. Later in 2002, the HFEA reached the opposite decision in the superficially similar case of Michelle and Jayson Whitaker. This paper will present a critical overview of the decisions of the HFEA in these two cases, and of the opinions and discussions the HFEA published in support thereof. Although it is acknowledged that the two cases differ to some extent in their facts, it is argued that the difference upon which the HFEA relied is one without ethical significance.     

PI3-kinase inhibitors abolish the enhanced chronotropic effects of angiotensin II in spontaneously hypertensive rat brain neurons

Angiotensin II (Ang II), acting at Ang II type 1 receptors (AT1Rs), increases the firing rate of neurons from Wistar-Kyoto (WKY) rat brain via protein kinase C (PKC)- and calcium-calmodulin kinase II (CaMKII)-dependent mechanisms. The objectives of this study were twofold; first, to compare the Ang-II-stimulated increase in firing of neurons from WKY and spontaneous hypertensive rats (SHR) and second, to elucidate the signaling mechanisms involved. Action potentials were measured in neurons cultured from SHR and WKY rat brains using the whole cell configuration of the patch-clamp technique in the current-clamp mode. Ang II (100 nM) caused three- and sixfold increases in neuronal firing rate in WKY rat and SHR neurons, respectively; effects that were abolished by the AT1R antagonist Losartan (1 microM). Co-administration of calphostin C (10 microM, a PKC inhibitor) and KN-93 (10 microM, a CaMKII inhibitor) completely blocked this Ang II action in WKY rat neurons, while they caused only a approximately 50% attenuation in SHR neurons. The residual increase in firing rate produced by Ang II in SHR neurons was blocked by inhibitors of phosphatidylinositol 3 kinase (PI3-kinase), either LY 294002 (10 microM) or wortmannin (100 nM). These observations suggest that a PI3-kinase signaling pathway may be responsible for the enhanced chronotropic effect produced by Ang II in SHR neurons.     

Investigations into the amino-terminal domain of the respiratory syncytial virus nucleocapsid protein reveal elements important for nucleocapsid formation and interaction with the phosphoprotein

Bacterially expressed nucleocapsid (N) protein, from respiratory syncytial virus (RSV), was used to investigate RNA binding in a modified North-Western blotting protocol. The recombinant protein demonstrated no sequence specificity in binding RNA representing either the antigenomic leader sequence or the nonspecific sequence derived from a plasmid vector. When recombinant N was purified on CsCl gradients, two types of structure, both with densities indicating that they contained RNA, could be visualised by negative-stain electron microscopy. Structures similar to nucleocapsids (NC) from RSV-infected cells were observed, as were ring structures. A small fragment of the N (amino acids 1-92) was all that was required for the production of NC-like structures. Another mutant with an internal deletion could form rings but not NC-like structures. This suggests that this domain (amino acids 121-160) may be important for maintaining helical stability. Further analysis has also identified a potential site in the amino-terminus that may be involved in an interaction with the phosphoprotein. A domain model of the RSV N protein is presented which, similar to that of other paramyxoviruses, supports the idea that the amino-terminus is important for NC assembly.     

Simplifying collection of corneal specimens in cases of suspected bacterial keratitis

Identification of the causative organisms in suspected bacterial keratitis traditionally involves collecting multiple corneal scrapes, which are plated directly onto different solid agar culture media. Difficulties have been reported with this practice, so the development of a simpler diagnostic method in suspected bacterial keratitis would be useful. It is unclear whether a single corneal scrape sent to the microbiology laboratory in a liquid transport culture medium (indirect method) is as reliable for the diagnosis of bacterial keratitis as inoculation of multiple scrapes directly onto agar plates (direct method). To investigate this, bacterial recovery was assessed following transfer and transport of different concentrations and types of bacteria from an artificially contaminated surgical blade into brain heart infusion (BHI). Bacterial recovery rates between the proposed (indirect) and standard (direct) method were then compared after the in vitro inoculation of pig corneas and following specimen collection in patients with presumed bacterial ulcerative keratitis. Recovery of bacteria from contaminated surgical blades was found to be the same from both solid and liquid culture media. There was no significant difference in the numbers of positive cultures from solid (direct) and liquid (indirect) culture media, both in the experimental pig cornea inoculation study (P = 0.34) and in experiments with patients with clinical infections (P = 0.4), with an 85.2% agreement between methods (kappa = 0.61, P < 0.0001). In conclusion, therefore, the collection of two corneal scrapes, one used for Gram staining and the other transported in BHI followed by plating and subculturing in an enrichment medium, provides a simple method for the investigation of presumed bacterial keratitis.     

A prospective study and review of pre-morbid characteristics in children with traumatic brain injury

The pre-morbid characteristics of 32 children requiring inpatient rehabilitation following moderate or severe head injuries were studied prospectively. Teachers completed Rutter and modified Conners questionnaires on the pre-head injury behavioural characteristics of each child and also rated the pre-morbid learning ability of the children on a four-point scale. The children's socioeconomic status was determined using their postcode at the time of the injury. Behavioural questionnaires were returned for 25 of the 32 children. Five of the 25 children obtained total behaviour problem scores on the Rutter teacher scales indicative of pre-morbid psychopathology. The general learning ability of five children was rated as being far below average. The children had a higher mean socioeconomic deprivation score than for the geographic area in which the rehabilitation unit is sited. The incidence of pre-morbid behaviour problems in the study population with moderate or severe traumatic brain injury may be somewhat higher than in the local population, but this did not appear to be a major risk factor in sustaining a head injury. However, adverse environmental conditions, linked to socioeconomic deprivation, did appear to be important risk factors that contributed to a pedestrian road traffic accident.     

Pregnancy and birth after assisted reproduction

Pregnancy and birth after IVF has attracted much critical attention. Overall, pregnancy rates are about 30-40% with three transferred embryos. Abortion rates are high before 19 weeks of gestation, at 8% of established pregnancies, and ectopic pregnancies are regular occurrences. Pregnancy rates are highest (35%) in women aged 20-24 years, declining to 10% at and above age 40. The use of intracytoplasmic sperm injection (ICSI) and blastocyst transfer has transformed the establishment of pregnancies for couples with differing indications. High rates of implantation are achieved with blastocyst transfers, although many embryos die in vitro before this stage. Twins and triplets can result in up to 50% of pregnancies. Heterotopic pregnancies are rare. Abortion afflicts fetuses before 12 weeks (vanishing fetuses). Rates of spontaneous abortion can be as high as 50% with quadruplets and quintuplets, and may be even higher in cases of fetal aneuploidy. At birth, prematurity is high, especially with twins or higher multiple pregnancies. Fetal reduction is used to reduce multiple pregnancies for singletons. Ovarian hyperstimulation is a risk associated with multiple pregnancy.     

The extraction of permanent second molars and its effect on the dentofacial complex of patients treated with the Tip-Edge appliance

The aim of this investigation was to assess the dentofacial changes in a group of patients consecutively treated with Tip-Edge appliances and the extraction of four permanent second molars by one specialist orthodontic practitioner. Before and after treatment lateral cephalograms and study cast measurements of 45 individuals, 26 females (mean age 13.8 years) and 19 males (mean age 13.9 years), were collated and statistically analysed. Cephalometric variables that exhibited, before treatment, significant sex differences, included SNA, SNB (both smaller in males, P < 0.05) and U1-NA degrees (P < 0.05), nasolabial angle (P < 0.05), and upper lip length P < 0.01 (all larger in males). After treatment, sex differences were demonstrated for SNA (smaller in males, P < 0.05), mandibular length (P < 0.01), upper face height (P < 0.05), lower face height (P < 0.01), anterior face height (P < 0.001), posterior face height (P < 0.01), nasolabial angle (P < 0.05), and upper lip length and thickness (P < 0.001; all larger in males). For the cast analysis, before treatment differences indicated larger values for males than females for lower arch inter-canine, premolar, and molar widths, arch depth (all P < 0.05), tooth size, and arch length (P < 0.01). Similar findings were noted in the upper arch except for inter-canine and premolar arch width. Despite most arch variables displaying sex differences, no gender effect was found for irregularity or crowding parameters. The same variables exhibited significant sex differences and changes after treatment (except tooth size, lower arch depth, and upper arch inter-canine width). Overall, the pattern of correction exhibited by the subjects included dental, skeletal, and soft tissue changes. Males tended to have greater mean increases in mandibular skeletal and soft tissue variables compared with females. Both males and females had increases in most dental arch variables measured from the study casts. Both sexes demonstrated a small uprighting, but statistically non-significant distalizing of the buccal segments. The lower incisors in the sagittal plane revealed a mean tendency to remain in their pre-treatment positions, with some individual variation. Overall, the treatment results were considered favourable, but case selection appeared to bias towards Angle Class I skeletal patterns of average to slightly reduced facial height, overbite and overjet < or = 4 mm, lip competence, no incisor protrusion, and moderate tooth size to arch length discrepancy (3-3.5 mm lower arch, 1 mm upper arch). Further evaluation of third molar eruption responses may provide insight into appropriate timing of second molar extractions.