Implication of lipid turnover for the control of energy balance.

The ongoing obesity epidemic is a consequence of a progressive energy imbalance. The energy-balance model (EBM) posits that obesity results from an excess in food intake and circulating fuels. A reversal in causality has been proposed recently in the form of the carbohydrate-insulin model (CIM), according to which fat storage drives energy imbalance. Under the CIM, dietary carbohydrates shift energy use in favour of storage in adipose tissue. The dynamics of lipid storage and mobilization could, therefore, be sensitive to changes in carbohydrate intake and represent a measurable component of the CIM. To characterize potential changes in lipid dynamics induced by carbohydrates, mathematical models were used. Here, we propose a coherent mathematical implementation of the CIM-energy deposition model (CIM-EDM), which includes lipid turnover dynamics. Using lipid turnover data previously obtained by radiocarbon dating, we build two cohorts of virtual patients and simulate lipid dynamics during ageing and weight loss. We identify clinically testable lipid dynamic parameters that discriminate between the CIM-EDM and an energy in, energy out implementation of the EBM (EBM-IOM). Using a clinically relevant two-month virtual trial, we additionally identify scenarios and propose mechanisms whereby individuals may respond differently to low-carbohydrate diets. This article is part of a discussion meeting issue 'Causes of obesity: theories, conjectures and evidence (Part II)'.

Adipose lipid turnover and long-term changes in body weight.

The worldwide obesity epidemic1 makes it important to understand how lipid turnover (the capacity to store and remove lipids) regulates adipose tissue mass. Cross-sectional studies have shown that excess body fat is associated with decreased adipose lipid removal rates2,3. Whether lipid turnover is constant over the life span or changes during long-term weight increase or loss is unknown. We determined the turnover of fat cell lipids in adults followed for up to 16 years, by measuring the incorporation of nuclear bomb test-derived 14C in adipose tissue triglycerides. Lipid removal rate decreases during aging, with a failure to reciprocally adjust the rate of lipid uptake resulting in weight gain. Substantial weight loss is not driven by changes in lipid removal but by the rate of lipid uptake in adipose tissue. Furthermore, individuals with a low baseline lipid removal rate are more likely to remain weight-stable after weight loss. Therefore, lipid turnover adaptation might be important for maintaining pronounced weight loss. Together these findings identify adipose lipid turnover as an important factor for the long-term development of overweight/obesity and weight loss maintenance in humans.

Analysis of 14C and 13C in teeth provides precise birth dating and clues to geographical origin.

The identification of human bodies in situations when there are no clues as to the person's identity from circumstantial data, poses a difficult problem to the investigators. The determination of age and sex of the body can be crucial in order to limit the search to individuals that are a possible match. We analyzed the proportion of bomb pulse derived carbon-14 ((14)C) incorporated in the enamel of teeth from individuals from different geographical locations. The 'bomb pulse' refers to a significant increase in (14)C levels in the atmosphere caused by above ground test detonations of nuclear weapons during the cold war (1955-1963). By comparing (14)C levels in enamel with (14)C atmospheric levels systematically recorded over time, high precision birth dating of modern biological material is possible. Above ground nuclear bomb testing was largely restricted to a couple of locations in the northern hemisphere, producing differences in atmospheric (14)C levels at various geographical regions, particularly in the early phase. Therefore, we examined the precision of (14)C birth dating of enamel as a function of time of formation and geographical location. We also investigated the use of the stable isotope (13)C as an indicator of geographical origin of an individual. Dental enamel was isolated from 95 teeth extracted from 84 individuals to study the precision of the (14)C method along the bomb spike. For teeth formed before 1955 (N=17), all but one tooth showed negative Δ(14)C values. Analysis of enamel from teeth formed during the rising part of the bomb-spike (1955-1963, N=12) and after the peak (>1963, N=66) resulted in an average absolute date of birth estimation error of 1.9±1.4 and 1.3±1.0 years, respectively. Geographical location of an individual had no adverse effect on the precision of year of birth estimation using radiocarbon dating. In 46 teeth, measurement of (13)C was also performed. Scandinavian teeth showed a substantially greater depression in average δ(13)C (-14.8) than teeth from subjects raised in Japan (-13.5), Middle East and North Africa (-12.7) and South America (-10.9). In summary, isotopic analysis of carbon in enamel from a single tooth can give a good estimate of the year of birth of an individual and also provide information about the geographical origin of the individual. This strategy can assist police and forensic authorities when attempting to solve unidentified homicide cases and may facilitate the identification work associated with mass disasters.

Year of birth determination using radiocarbon dating of dental enamel.

Radiocarbon dating is typically an archaeological tool rather than a forensic one. Recently however, we have shown that the amount of radiocarbon present in tooth enamel, as a result of nuclear bomb testing during the cold war, is a remarkably accurate indicator of when a person is born. Enamel isolated from human teeth is processed to form graphite and carbon-14 ((14)C) levels are measured using accelerator mass spectrometry. Since there is no turnover of enamel after it is formed, (14)C levels in the enamel represent (14)C levels in the atmosphere at the time of its formation. In this paper we describe the strategy used to determine the date of birth of an individual based on radiocarbon levels in tooth enamel, focusing on the methodology of this strategy. Year of birth information can significantly assist police investigators when the identity of a deceased individual is unknown. In such cases police will try to match particulars of the unidentified individual (which is often only gender and/or an estimate of age), with particulars from missing persons lists.

Retinal ganglion cell neurotrophin receptor levels and trophic requirements following target ablation in the neonatal rat.

Superior colliculus (SC) ablation in neonatal rats results in a rapid increase in retinal ganglion cell (RGC) death. This injury-induced death is reduced by exogenous brain-derived neurotrophic factor or neurotrophin-4/5 (NT-4/5), but the protective effect of these molecules is transient, delaying but not preventing neuronal loss. We sought to discover why neurotrophins only temporarily reduce RGC death after target ablation, focusing on changes in neurotrophin receptor expression and possible changes in growth factor dependency. In unlesioned rats, receptor tyrosine kinase B (trkB) immunohistochemistry revealed no change in the number of trkB positive cells in the RGC layer 24 h after intraocular NT-4/5 injection. However, after SC lesions there were significantly less immunoreactive cells and, surprisingly, even fewer immunoreactive cells in NT-4/5 injected eyes. Semi-quantitative confocal analysis of immunofluorescence intensity revealed an increase in trkB staining in the RGC layer in unlesioned rats 24 h after NT-4/5 injection, whereas in SC-lesioned animals exposed to NT-4/5 there was a significant decrease in staining. To determine whether injured neonatal RGCs can switch their trophic requirements, different doses of ciliary neurotrophic factor were given intraocularly, either alone or combined with NT-4/5. We also tested an SC-derived chondroitin sulfate proteoglycan that has been reported to promote neonatal RGC survival. None of these interventions reduced lesion-induced RGC death 24 or 36 h after SC ablation. In summary, we show that developing RGCs do not shift their trophic dependence to other survival factors following injury; rather, the application of neurotrophins causes a down-regulation of the cognate trkB receptor, presumably altering the long-term responsiveness of neonatal RGCs to exogenous neurotrophins. These data highlight the difficulty in promoting long-term neuronal survival when using one-off administration of recombinant growth factors.

The effects of central administration of neurotrophins or transplants of fetal tectal tissue on retinal ganglion cell survival following removal of the superior colliculus in neonatal rats.

In neonatal rats, intraocular injections of brain-derived neurotrophic factor (BDNF) or neurotrophin 4/5 (NT-4/5) enhance the survival of retinal ganglion cells (RGCs) following superior colliculus (SC) ablation [Q. Cui, A.R. Harvey, At least two mechanisms are involved in the death of retinal ganglion cells following target ablation in neonatal rats, J. Neurosci., 15, 1995, pp. 8143-8155.]. The aim of the present study was to determine if: (i) fetal tectal tissue grafted into the lesion site, or (ii) neurotrophins applied centrally to the injured SC, also decreased lesion-induced RGC death. Nuclei of tectally projecting RGCs were identified by injecting diamidino yellow (DY) into the left SC of 2-day-old (P2) Wistar rats. Injected SCs were lesioned at P4. In some animals, embryonic (E16) tectal tissue was then implanted into the lesion cavity; host rats were perfused 24 h or 20 days later. In short-term (24-h) studies, the number of DY-labelled pyknotic profiles was compared to the number of normal DY-labelled RGCs in retinal wholemounts (right eyes). The proportion of dying RGCs in animals with grafts (10.7%, n = 17) was not significantly different from lesion-only rats (13.2%, n = 26). Nonetheless, the long-term (20-day) study showed that, in most rats, fetal tectal tissue survived in the lesion cavity and in some cases, the grafts received host retinal input. In another group, different doses of BDNF or NT-4/5 were applied to the SC after P4 tectal lesions. Rats were perfused 24 h later and the number of pyknotic vs. normal DY-labelled RGCs was determined. Initial trials in which SC lesions were filled with gelfoam soaked in BDNF or NT-4/5 were unsuccessful; however, RGC death was reduced (p < 0.05, Dunnett's test) in rats that received gelfoam implants as well as focal neurotrophin injections into SC rostral to the lesion. The lowest pyknotic rate in individual animals from the BDNF and NT-4/5 groups was 2.41% and 2.01%, respectively. Overall, the proportion of dying RGCs was 7.0% (n = 8) for BDNF and 7.4% (n = 17) for NT-4/5 treated rats. Normal RGC densities were also significantly higher in these animals. NT-4/5 topically applied to the posterior surface of the eye did not reduce RGC death. The data show that the viability of injured neonatal RGCs is increased by specific retrograde neurotrophin-mediated survival signals which can be activated from the SC.