Changes in some properties of glutamate decarboxylase activity during the maturation of the brain

The sensitivity of cerebral glutamate decarboxylase (GAD) activity to hypotonic homogenization medium, centrifugation, Triton-X-100, and preincubation at 37°C was studied in the developing mouse. In newborn and 5-day-old animals, GAD activity was markedly inhibited by all these conditions. From 5 days to adult age, the sensitivity of the enzyme to the experimental conditions used decreased progressively, with the greatest change between the 10th and 15th days. It is concluded that the newborn form of the enzyme, which is unstable and shows a relatively high affinity for pyridoxal phosphate, is substituted by the adult form during the maturation of the brain. The activity of the adult form is much more stable and more dependent on free pyridoxal phosphate. The implications of these findings in the regulation of cerebral excitability during development are discussed.     

THE TECHNIQUE OF LABELLED MITOSES: ANALYSIS BY AUTOMATIC CURVE-FITTING

An improved method is described for the analysis of data obtained by the technique of labelled mitoses. It is a development of the method described by Barrett (1966) in which theoretical curves are computed on the basis of a model which assumes that the phases G¹, S and G² are described by independent log-normal distributions; the analysis consists in finding a form of this model which gives a labelled mitoses curve which is the best fit to the available data. This fitting procedure has now been made automatic. No comprehensive indication of the goodness of fit can be given, although in the analysis of over fifty sets of data the method appears to have worked well.
A supplementary computer program is described which, on the basis of three separate assumed modes of cell loss, calculates the form of the age distributions and theoretical continuous labelling curves. This allows growth fraction to be calculated in a way which takes account of the distribution of phase durations and the non-rectangular age distributions of expanding cell populations. It also gives an opportunity to study the implications of continuous labelling data as regards the mode of cell loss.
A comparison is made between the present method of labelled mitoses curve analysis and the empirical rules which have often been used.     

Quantitative analysis of the temporal pattern of somite formation in the mouse and rat. A simple and accurate method for age determination

The relation between the logarithm of the volume of the embryo and the number of somites was studied in the mouse and rat in the period of about 0-43 somites, a period characterized by a very rapid volume increase. A linear relation was found, and the variability of the individual observations around the straight line was rather small. Using a previously defined age estimate, i.e. the 'developmental age' based on the embryonic volume, we also found a linear relation between this age estimate and the number of somites. With the help of this straight line, the developmental age can be estimated from the number of somites with an accuracy of less than 0.1 day. Therefore, the developmental age can be considered a better age estimate than those based on the usual methods. The time needed to form a new somite was found to be constant in both species during the period studied. The length of the period differed, and was 1.68 h in the mouse and 2.24 h in the rat. However, the volume of the embryo at the time when a specific somite was formed (for instance the 25th) appeared to be the same in both species. Comparison of the volume of embryos, embedded in paraffin and Epon and sectioned, enabled us to estimate the shrinkage in Epon embryos. The volume after processing was approximately 62% of the original volume.     

Latest developments of dendro-archaeology in the French Alps: Assessing changes in mountain forests exploitation over the last millennium

Dendrochronological studies are carried out on rural buildings in the French Alps for twenty years. In this contribution, we examine an extensive dataset of larch timbers (n = 1294) that were dendrochronologicaly dated at 139 buildings. This material yielded felling dates spanning the 11th century to the 20th century and contributed to improve our knowledge of human occupation and exploitation of mountain forests during the second millennium CE. From the 11th to the 14th century, rural populations increasingly exploited older forests. The crises of the 14th century led to a significant decrease in construction and by extension in tree cutting. The beginning of the 15th century marks a resumption of construction and felling which is continuous until the beginning of the 20th century. There is a relationship between the age of harvested trees, the number of dates and population fluctuations, which corroborates findings at the European scale. Regional trade probably influenced the stem diameters used in constructions in the immediate vicinity of the forests. Logging regulations can be perceived through variations in the age of harvested trees. The 14th century gap in felling dates would also mark a transition period in the exploitation of mountain forests and by extension a transformation of the economy.     

Assembly of the precursor and processed light-harvesting chlorophyll a/b protein of Lemna into the light-harvesting complex II of barley etiochloroplasts

When the in vitro synthesized precursor of a light-harvesting chlorophyll a/b binding protein (LHCP) from Lemna gibba is imported into barley etiochloroplasts, it is processed to a single form. Both the processed form and the precursor are found in the thylakoid membranes, assembled into the light-harvesting complex of photosystem II. Neither form can be detected in the stromal fraction. The relative amounts of precursor and processed forms observed in the thylakoids are dependent on the developmental stage of the plastids used for uptake. The precursor as well as the processed form can also be detected in thylakoids of greening maize plastids used in similar uptake experiments. This detection of a precursor in the thylakoids, which has not been previously reported, could be a result of using rapidly developing plastids and/or using an heterologous system. Our results demonstrate that the extent of processing of LHCP precursor is not a prerequisite for its inclusion in the complex. They are also consistent with the possibility that the processing step can occur after insertion of the protein into the thylakoid membrane.     

Free radicals and disease.

Free radicals and reactive oxygen species (ROS) have been associated with the etiology and/or progression of a number of diseases and in aging. Many of the proteins oxidatively modified by free radicals contain side-chain carbonyl derivatives, which can be used as markers for protein oxidation. The protein carbonyl content has been quantitated as a function of age for human cultured dermal fibroblasts, lens, and brain tissue. These data were analyzed using a simple autocatalytic model with the assumption that free radicals randomly oxidize proteins or peptides to form carbonyl derivatives and lead to their inactivation. The carbonylated proteins and peptides are highly susceptible to proteolytic degradation. Implication of free radicals in aging and in age-dependent susceptibility to neurodegenerative diseases will be discussed in light of this simplified kinetic model.     

Juvenile/total foliage ratios inEucalyptus nitens and the growth of stands and individual trees

Summary Individual trees and stands of two provenances of Eucalyptus nitens which have marked differences in retention of juvenile foliage were studied in four plantations at different elevations. The proportion of juvenile to total foliage and growth was measured at the end of the 2nd, 3rd and 4th year from establishment. Between the ages of 2 and 4 years annual stem volume increment increased in proportion to the amount of juvenile foliage retained. By age 4 years, stem volume of trees of the juvenile persistent form was significantly larger than that of the early adult form. Increasing differences in height growth with age between provenances, which were highly significant across sites by age 4, contributed to these differences in performance. There was some evidence that the largest trees of the juvenile-persistent form were those which combined mature foliage above juvenile foliage for the longest period during the transition from juvenile to mature foliage. In the early-adult form the largest trees were those which completed the transition to mature foliage rapidly. There was no difference in the ratio of foliage mass to basal area between the two forms. It is suggested that the faster growth of the juvenile-persistent form is related to higher leaf area index and not to foliage type. A provenance of E. globulus which had a higher retention of juvenile foliage at age 4 than a second provenance had a lower stem volume, thus indicating that in this species early growth rate is not determined by foliage type.     

Meckel the Younger and his epistemology of organic form: Morphology in the pre-Gegenbaurian age

Summary The goal of our paper is to investigate Meckel’s epistemology of organic form, based on study of his original publications. Johann Friedrich Meckel the Younger (1781–1833) was one of the leading figures of German morphology in the early 19th century. Historiographic studies on morphology in this time period show, that biological research was largely preoccupied with questions about the relationship between form and function. Investigations into Meckel’s epistemology of organic form can contribute to our understanding of the development of morphology in the pre-Gegenbaurian age.     

Virilizing effect of methyltestosterone on female descendants in the rat

Pregnant rats were given daily a subcutaneous injection of methyltestosterone for 4 days from the 17th to the 20th day of gestation, and were allowed to be delivered to their offsprings (F1) which were used for the examination of later reproductive functioning. When observed for 21 weeks after birth, the growth rate of F1 from methyltestosterone-treated groups was higher than that of F1 from the control group. The anogenital distance in 50-microgram-treated F1 females started to become significantly longer on the 14th day and in 5-microgram-treated F1 females on the 28th day after birth than that in F1 from the control. The day on which vaginal opening took place in 50% of females was 34.4 days of age in both the control and the 5 microgram groups, but it delayed until 40.7 days in the 50 microgram group. Furthermore, persistent estrus was observed after about 90 days of age in the 50 microgram group. This persistent estrus disappeared by placing these females with males, resulting no pregnancy. In the 5 microgram group females could be pregnant, but their female fetuses (F2), when examined on the 21st day of gestation, had significantly shortened the length of the urovaginal septum. The observations show that virilization can be induced in the third generation.     

Electrocorticographic development of epileptogenic foci in the occipital region of the rat cerebral cortex

The development of cortical penicillin foci in the occipital region was studied in rats whose ages ranged from five days up to the adult age. The local application of penicillin induced the formation of an epileptogenic focus for the first time at the age of seven days. With advancing age, the amplitude of focal discharges increased, the duration of the individual components of the discharge shortened, its originally negative-positive configuration changed to a triphasic form and in the third week of life initial positivity, for a time, become the dominant component of the discharge. Projection of the discharges to the contralateral hemisphere was found to be inconstant in the second postnatal week, but appeared regularly from the age of 14 days. Synchronization of the discharges of two symmetrical foci was very poor in 7-day-old young, but improved noticeably by the 14th day; it was never complete, however, even in adulthood. The activity of symmetrical foci changed spontaneously to ECoG seizures, which were most common in 7-day-old young (in which ictal activity was usually not generalized, however) and were least frequent in 14-day-old animals. Focal discharges could not be reliably triggered by electrical stimulation of the contralateral cortex until the age of 18 days and later. The occipital part of the cortex develops somewhat later than the sensorimotor, frontal region, and during its development there also appeared phenomena which are not present in the frontal cortex.     

Purification and characterization of rabbit muscle acylphosphatase in the thiol (-SH) form

Modifications in the muscle acylphosphatase purification procedure enabled us to isolate the enzyme with its sole cysteine in the -SH form; this enzyme form is the most abundant in vivo. Our data demonstrates that the enzyme forms purified by previously reported procedures can be easily derived from a reaction of the SH-enzyme with oxidized glutathione. Probably most, or even all, of these enzyme forms are artifacts due to the purification. The SH-acylphosphatase shows kinetic parameters similar to those reported for the mixed disulfide with glutathione and S-S dimer, except for the specific activity value, which is about twice as much, and the Km, which is reduced.     

Immunocytochemical study using semi-thin sections of the phenomena of early differentiation in several cell populations of the human fetal adenohypophysis]

Immunocytological investigations have been performed on semi-thin sections of human fetal pituitaries ranging in fetal age from 6 to 26 weeks. Corticotrophs can be revealed by anti-ACTH (1-24), anti-ACTH (17-39) and anti-beta MSH but not by anti-alpha MSH immunesera from the 8th week. Somatotrophs are revealed with anti-human STH from 9th week. Differentiating cells containing only alpha subunits of glycoproteic hormones are present from 8th to 12th week. At 13th week beta subunits of TSH can be revealed immunocytologically in thyreotrophs. Beta subunits of LH or FSH can be detected in same gonadotrophic cells only from 15th week.     

Recent histories of six productive lakes in the Irish Ecoregion based on multiproxy palaeolimnological evidence

Palaeolimnological data from six mesotrophic, eutrophic and hypertrophic lakes in the Irish Ecoregion, in the form of microfossil (cladocera, diatoms and pollen) and sediment chemistry data from radiometrically dated sediment cores, were used to reconstruct past variations in lake water quality and catchment conditions. Basal sediments from sediment cores from the six sites ranged in age from the late 18th century to the early 20th century. A weighted averaging partial least squares regression model was developed to reconstruct past epilimnetic total phosphorus concentrations. The results indicate that all but one of the study sites currently are in a far more productive state compared with the beginning of the sediment core record and that those same five lakes have experienced accelerated enrichment post c. 1980. Two of the sites demonstrated long-term enrichment, in one case beginning in the late 19th century, while both eutrophication and oligotrophication have occurred at three sites. The results highlight the difficulties in applying a general temporal end-point for reference conditions and demonstrate that productive lakes in the Irish Ecoregion have complex, locally specific and often long histories of enrichment. These may not be responsive to reduced external loadings of phosphorus and, as a result, restoration could prove particularly challenging. The results also provide evidence of the ways in which palaeolimnological techniques can assist implementation of the EU Water Framework Directive. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

The effect of microwaves on the neuronal activity of the hyperstriatum in chick embryos at the critical developmental period]

Studies have been made on multineuronal activity in the dorsal hyperstriatum of chick embryos exposed to microwave radiation (2.375 MHz) at critical stage of development of the structure (19th day of incubation). One day after microwave exposition, activation of neurones was observed in a form of the increase in the discharge frequency and the increase of outburst form of the activity. Cyclic form of outburst activity was absent in experimental animals being present in control embryos of the same age and associated presumably with self-organization of microsystems of neurones in the developing brain. Changes in multineuronal activity were found only in the left hyperstriatum, which is probably due to different levels of the development and functional activity of the right and left hyperstriatum at this stage resulting from asymmetric intensity of visual afferentation which depends on the position of embryo inside the egg.     

Using stable isotope analysis to examine the effect of economic change on breastfeeding practices in Spitalfields, London, UK

Breastfeeding patterns were subject to a number of fads in 18th and 19th century Britain. Feeding infants by hand, rather than maternal breastfeeding or wet-nursing, became more prevalent among both the wealthy and poor. Substitute foods may have been a convenient alternative for mothers employed away from the household. This study used stable isotope ratio analysis to examine the weaning schedule in the 18th and 19th century skeletal assemblage from Spitalfields, London, UK. Analysis of 72 juvenile ribs revealed δ(15) N elevations of 2-3‰ above the adult mean for individuals up to the age of two, while elevations of 1-2‰ were observed in δ(13) C for the first year of life. This suggests that the introduction of solid foods took place before the end of the first year, and that breastfeeding had entirely ceased by 2 years of age. The age at death of many of these infants is known from historical records, and can be used to pinpoint the amount of time required for the breast milk signal to be observed in the stable isotope ratios of rib collagen. Results show that a δ(15) N elevation can be detected in the ribs of individuals who died as young as 5-6 weeks. Not all individuals at Spitalfields were breastfed, and there may not have been a single uniformly practiced weaning scheme. There is, however, more evidence for prolonged breastfeeding during the 19th century than the 18th century.     

Redox potentials of the photosynthetic bacterial cytochromes c2 and the structural bases for variability.

The cytochromes c2 of the Rhodospirillaceae show a much greater variation in redox potential and its pH dependence than the mitochondrial cytochromes c that have been studied. It is proposed that the range of redox potential for cytochromes c2 functioning as the immediate electron donor to photo-oxidised bacteriochlorophyll may be 345-395 mV at pH 5. Closely related cytochromes c2 with different redox potentials show patterns of amino acid substitution which are consistent with changes in hydrophobicity near the haem being at least a partial determinant of redox potential. More distantly related cytochromes are difficult to compare because of the large number of amino acid substitutions and the probability that there are subtle changes in overall peptide chain folding. The redox potential versus pH curves can be analysed in terms of either one ionisation in the oxidised form or two in the oxidised form and one in the reduced. The pK in the oxidised form at higher pH values can be correlated with the pK for the disappearance or shift of the near infrared absorption band located near 695 nm. The structural bases of these ionisations are not known but the possible involvement of the haem propionate residues is discussed.     

Risk prediction of complex diseases from family history and known susceptibility loci, with applications for cancer screening

Risk prediction based on genomic profiles has raised a lot of attention recently. However, family history is usually ignored in genetic risk prediction. In this study we proposed a statistical framework for risk prediction given an individual's genotype profile and family history. Genotype information about the relatives can also be incorporated. We allow risk prediction given the current age and follow-up period and consider competing risks of mortality. The framework allows easy extension to any family size and structure. In addition, the predicted risk at any percentile and the risk distribution graphs can be computed analytically. We applied the method to risk prediction for breast and prostate cancers by using known susceptibility loci from genome-wide association studies. For breast cancer, in the population the 10-year risk at age 50 ranged from 1.1% at the 5th percentile to 4.7% at the 95th percentile. If we consider the average 10-year risk at age 50 (2.39%) as the threshold for screening, the screening age ranged from 62 at the 20th percentile to 38 at the 95th percentile (and some never reach the threshold). For women with one affected first-degree relative, the 10-year risks ranged from 2.6% (at the 5th percentile) to 8.1% (at the 95th percentile). For prostate cancer, the corresponding 10-year risks at age 60 varied from 1.8% to 14.9% in the population and from 4.2% to 23.2% in those with an affected first-degree relative. We suggest that for some diseases genetic testing that incorporates family history can stratify people into diverse risk categories and might be useful in targeted prevention and screening.     

The Archaeal Concept and the World it Lives in: A Retrospective

The present retrospective concerns the discovery and development of the archaea, the so-called 'third form of life' that no one anticipated and many did not, and still do not want. In its birth pangs, which the archaea had a plenty, the concept encountered biology unmasked; for it ran up against some of the key struts in the 20th century biological edifice. Consequently, the history of the development of the archaeal concept provides an excellent window on certain of the weaknesses in the 20th century biology paradigm, weaknesses that have now led that paradigm to a conceptual dead end. On the other hand, the archaeal concept has also provided us one of the pillars on which a new holistic paradigm for biology can be built. So, it would seem of value to retrace some of the twists and turns in the history of the development of the archaeal concept. Given my position vis-à-vis the archaea, my account will be a personal one.     

A perspective on mechanisms of protein tetramer formation

Homotetrameric proteins can assemble by several different pathways, but have only been observed to use one, in which two monomers associate to form a homodimer, and then two homodimers associate to form a homotetramer. To determine why this pathway should be so uniformly dominant, we have modeled the kinetics of tetramerization for the possible pathways as a function of the rate constants for each step. We have found that competition with the other pathways, in which homotetramers can be formed either by the association of two different types of homodimers or by the successive addition of monomers to homodimers and homotrimers, can cause substantial amounts of protein to be trapped as intermediates of the assembly pathway. We propose that this could lead to undesirable consequences for an organism, and that selective pressure may have caused homotetrameric proteins to evolve to assemble by a single pathway.     

Effect of thyroxine administration to the mother on postnatal radioiodine uptake by the thyroid of partially thyroidectomized rats.

The experiment was carried out on 35 litters of infant rats aged 4-17 days. The animals in each litter were always divided into two groups: control (sham operation) and experimental (hemithyroidectomy). Starting with the day on which the young were operated on, the mothers received daily subcutaneous injections of either saline or of thyroxine in doses of 50, 100 or 200 mug. At the end of the experiment, the young were injected intraperitoneally with 1 muCi 131I. One hour later they were decapitated and the radioactivity in their thyroid was expressed as the percentage of the administered dose per mg thyroid. The following age groups were used, according to the interval between thyroidectomy and decapitation: 4 to 8, 9 to 13, 13 to 15 and 15 to 17 days. 131I uptake by the residue of the thyroid in partially thyroidectomized animals was always compared with the values in the animals from the same litter subjected to sham operation. The results showed that partial thyroidectomy significantly stimulated 131I uptake in all age groups in which the mother was only given saline. In the 4- to 8-day-old group, the administration of 50 or 100 mug thyroxine to the mother inhibited this compensatory increase. In the 9- to 13-day-old group, inhibition occurred only after a dose of 100 mug thyroxine. In animals with an interval from the 13th to the 15th days old the dose of thyroxine administered to the mother had to be raised to 200 mug/day to achieve an inhibitory effect. In the last group (interval 15th to 17th day), not even administration of the maximum thyroxine dose to the mother from the 13th postnatal day succeeded in inhibiting the significant increase in 131I uptake. These results show that thyroxine administered to lactating female rats can be transmitted via the milk to the organism of the young in amounts which can be demonstrated in a physiological tests.