Variation in the strength of allometry drives rates of evolution in primate brain shape.

Large brains are a defining feature of primates, as is a clear allometric trend between body mass and brain size. However, important questions on the macroevolution of brain shape in primates remain unanswered. Here we address two: (i), does the relationship between the brain size and its shape follow allometric trends and (ii), is this relationship consistent over evolutionary time? We employ three-dimensional geometric morphometrics and phylogenetic comparative methods to answer these questions, based on a large sample representing 151 species and most primate families. We found two distinct trends regarding the relationship between brain shape and brain size. Hominoidea and Cercopithecinae showed significant evolutionary allometry, whereas no allometric trends were discernible for Strepsirrhini, Colobinae or Platyrrhini. Furthermore, we found that in the taxa characterized by significant allometry, brain shape evolution accelerated, whereas for taxa in which such allometry was absent, the evolution of brain shape decelerated. We conclude that although primates in general are typically described as large-brained, strong allometric effects on brain shape are largely confined to the order's representatives that display more complex behavioural repertoires.

Simultaneous detection of macroevolutionary patterns in phenotypic means and rate of change with and within phylogenetic trees including extinct species.

Recognizing evolutionary trends in phenotypic means and rates requires the application of phylogenetic comparative methods (PCMs). Most PCMs are unsuited to make full use of fossil information, which is a drawback, given the inclusion of such data improves, and in some cases even corrects, the proper understanding of trait evolution. Here we present a new computer application, written in R, that allows the simultaneous computation of temporal trends in phenotypic mean and evolutionary rate along a phylogeny, and to contrast such patterns among different clades within the tree. By using simulation experiments, we show the new implementation, names search.trend is as powerful as existing PCM tools in discerning macroevolutionary patterns in phenotypic means and rates, but differently from any other PCM allows comparing individual clades to each other, and provides rich information about trait evolution for all lineages in the tree.

Unexpectedly rapid evolution of mandibular shape in hominins.

Members of the hominins - namely the so-called 'australopiths' and the species of the genus Homo - are known to possess short and deep mandibles and relatively small incisors and canines. It is commonly assumed that this suite of traits evolved in early members of the clade in response to changing environmental conditions and increased consumption of though food items. With the emergence of Homo, the functional meaning of mandible shape variation is thought to have been weakened by technological advancements and (later) by the control over fire. In contrast to this expectation, we found that mandible shape evolution in hominins is exceptionally rapid as compared to any other primate clade, and that the direction and rate of shape change (from the ape ancestor) are no different between the australopiths and Homo. We deem several factors including the loss of honing complex, canine reduction, and the acquisition of different diets may have concurred in producing such surprisingly high evolutionary rates. This study reveals the evolution of mandibular shape in hominins has strong morpho-functional and ecological significance attached.

Pilot Study on Epigenetic Response to A Mind-Body Treatment.

In the last years, epigenetics and functional genomics methods to evaluate the genomic effects and mechanisms of mind-body therapies have increasingly grown. DNA microarray technology has been used to show the involvement of the stress response pathways both in the case of disease and stress and as an effect of mind-body therapies. In the present research, the DNA samples obtained from 20 individuals who experienced a mind-body therapeutic protocol (MBT-T), were analysed from the bio-molecular point of view by means of an epigenetic marker (MSAP molecular tool), in order to estimate the different status of methylation. The subjects were compared at 3 different times: prior to, 1 hour after, and 24 hours after the treatment. The molecular data were processed through different biostatistics approaches: the Bayesian statistics approach, in order to estimate the clustering membership of the subjects (Structure), and the statistical estimation of the DNA methylation level (MSAP statistical tool). The structure analysis revealed that the clusters and their membership changed among the three time points moving from higher heterogeneous distribution to higher homogeneous clusters. Before the treatment, the subjects' epigenetic profiles were heterogeneous; after the mind-body treatment we found that epigenetic profiles converged to homogeneous DNA methylation status. DNA epigenetic status of the subjects was affected by the MBT-T treatment.

Pilot Study on Epigenetic Response to A Mind-Body Treatment.

In the last years, epigenetics and functional genomics methods to evaluate the genomic effects and mechanisms of mind-body therapies have increasingly grown. DNA microarray technology has been used to show the involvement of the stress response pathways both in the case of disease and stress and as an effect of mind-body therapies. In the present research, the DNA samples obtained from 20 individuals who experienced a mind-body therapeutic protocol (MBT-T), were analysed from the bio-molecular point of view by means of an epigenetic marker (MSAP molecular tool), in order to estimate the different status of methylation. The subjects were compared at 3 different times: prior to, 1 hour after, and 24 hours after the treatment. The molecular data were processed through different biostatistics approaches: the Bayesian statistics approach, in order to estimate the clustering membership of the subjects (Structure), and the statistical estimation of the DNA methylation level (MSAP statistical tool). The structure analysis revealed that the clusters and their membership changed among the three time points moving from higher heterogeneous distribution to higher homogeneous clusters. Before the treatment, the subjects' epigenetic profiles were heterogeneous; after the mind-body treatment we found that epigenetic profiles converged to homogeneous DNA methylation status. DNA epigenetic status of the subjects was affected by the MBT-T treatment.

Progress to extinction: increased specialisation causes the demise of animal clades.

Animal clades tend to follow a predictable path of waxing and waning during their existence, regardless of their total species richness or geographic coverage. Clades begin small and undifferentiated, then expand to a peak in diversity and range, only to shift into a rarely broken decline towards extinction. While this trajectory is now well documented and broadly recognised, the reasons underlying it remain obscure. In particular, it is unknown why clade extinction is universal and occurs with such surprising regularity. Current explanations for paleontological extinctions call on the growing costs of biological interactions, geological accidents, evolutionary traps, and mass extinctions. While these are effective causes of extinction, they mainly apply to species, not clades. Although mass extinctions is the undeniable cause for the demise of a sizeable number of major taxa, we show here that clades escaping them go extinct because of the widespread tendency of evolution to produce increasingly specialised, sympatric, and geographically restricted species over time.

Clonal differences in survival capacity, copper and zinc accumulation, and correlation with leaf polyamine levels in poplar: a large-scale field trial on heavily polluted soil.

Three ex situ collections of poplar clones from natural populations of Populus alba and P. nigra growing in northern Italy were assessed for their genetic dissimilarity (GD) by means of amplified fragment length polymorphism (AFLP). The high GD evidenced within populations was exploited for screening 168 clones in a field trial on heavy metal-polluted soil. After one growth season, clonal differences in plant survival and growth were observed. On the basis of performance, six clones were singled out, and used to evaluate copper and zinc accumulation in different organs. Clonal differences in metal concentrations were most evident for leaves and stems; one clone of P. alba (AL35) had a distinctly higher concentration of both metals in the roots. Leaf polyamine (putrescine, spermidine, spermine) profiles correlated with tissue metal concentrations, depending on the clone, plant organ and metal. In particular, the high metal-accumulating clone AL35 exhibited a dramatically higher concentration of free and conjugated putrescine. Overall, the results indicate that, given the high GD of Populus even within populations, it is possible to identify genotypes best suited for soil clean-up, and useful also for investigating physiological markers associated with high metal accumulation/tolerance.

Ex-situ conservation of Black poplar in Europe: genetic diversity in nine gene bank collections and their value for nature development.

Populus nigra L. is a pioneer tree species of riparian ecosystems that is threatened with extinction because of the loss of its natural habitat. To evaluate the existing genetic diversity of P. nigra within ex-situ collections, we analyzed 675 P. nigra L. accessions from nine European gene banks with three amplified fragment length polymorphism (AFLP) and five microsatellite [or simple sequence repeat (SSR)] primer combinations, and 11 isozyme systems. With isozyme analysis, hybrids could be detected, and only 3% were found in the gene bank collection. AFLP and SSR analyses revealed effectively that 26% of the accessions were duplicated and that the level of clonal duplication varied from 0% in the French gene bank collection up to 78% in the Belgian gene bank collection. SSR analysis was preferred because AFLP was technically more demanding and more prone to scoring errors. To assess the genetic diversity, we grouped material from the gene banks according to topography of the location from which the accessions were originally collected (river system or regions separated by mountains). Genetic diversity was expressed in terms of the following parameters: percentage of polymorphic loci, observed and effective number of alleles, and Nei's expected heterozygosity or gene diversity (for AFLP). Genetic diversity varied from region to region and depended, to some extent, on the marker system used. The most unique alleles were identified in the Danube region (Austria), the Rhône region (France), Italy, the Rijn region (The Netherlands), and the Ebro region (Spain). In general, the diversity was largest in the material collected from the regions in Southern Europe. Dendrograms and principal component analysis resulted in a clustering according to topography. Material from the same river systems, but from different countries, clustered together. The genetic differentiation among the regions (F(st)/G(st)) was moderate.

Molecular analysis of natural populations of Populus nigra L intermingled with cultivated hybrids.

In this study six simple sequence repeats (SSR or microsatellites) were selected for their ability to fingerprint a total of 60 commercial clones of Populus deltoides Marsh. and Populus x canadensis Moench (typically derived from crosses between Populus nigra L and P. deltoides) and to characterize a natural population of P. nigra growing along the Ticino river in the North of Italy. Out of six SSRs used, four microsatellite loci were found to have alleles which were species-specific to P. deltoides and could therefore be used as markers for introgression of P. deltoides into P. nigra. In the studied region hybrid poplars and P. deltoides commercial clones are cultivated as monoclonal stands close to the area where black poplar has its natural habitat. SSR analysis was performed to investigate whether there was evidence of introgression between the natural population and the monoclonal plantations of hybrids and P. deltoides clones cultivated in the surrounding area. Three stages of the natural population were analysed: a group of old trees about a hundred years old, a younger population (aged 2-30 years) and the seedlings of three females of this population. Alleles specific to P. deltoides were detected only in the old cohort of the natural population, while no introgression was observed in the younger individuals and their progenies. These results were also confirmed by isozyme analysis of loci PGI-B, PGM and LAP-A, which were previously identified as diagnostic for P. nigra, P. deltoides and P.xcanadensis.

[A case of severe malnutrition due to Ascaris lumbricoides infestation in a boy]. / Su di un caso di grave malnutrizione da infestazione da Ascaris lumbricoides in un bambino.

The authors report a case of a child three years old, with severe malnutrition as complication of Ascaris lumbricoides infection. Intestinal nematodes infect many of the world's children and constitute a formidable public health problem. The infected children may suffer nutritional deficits, serious illness and occasionally death. Although infestation is uncommon in our country, it should be considered in children with low social life.

[Allergy to cow's milk proteins: the authors' personal cases (clinical characteristics, diagnostic methodology and prospective follow-up)]. / Allergia alle proteine del latte vaccino: casistica personale (caratteristiche cliniche, metodologia diagnostica, follow-up prospettico).

Because of specific laboratory tests are lacking, diagnosis of cow's milk allergy is always made on the basis of improvement after cow's milk protein withdrawal from diet and relapsed after challenge test. However personal and familial anamnesis, supported by few simple laboratory tests (peripheral blood and stool eosinophiles, hemoccult), are important tools for diagnosis. In this work we report the clinical findings of 68 children, suffered from cow's milk allergy, observed in the last 10 years. Children were divided into three groups on the basis of challenge response: Group 1 consisting of children with averse reaction occurred within the first hour after the administration of cow's milk protein (IgE-mediated reaction, 1st class as Gell-Coombs classification); Group 2 consisting of children with averse reaction occurred between 2nd and 12th hour (1st-3rd class as Gell-Coombs classification); Group 3 consisting of children with averse reaction occurred after 24 hours the administration of cow's milk proteins (4th class as Gell-Coombs classification). Auxological parameters show that while weight is widely involved, particularly in children of group 3, height is interested only in children with prolonged diarrhoea. Cow's milk protein withdrawal from diet determine a rapid normalization of weight increment rate, more evident in children over 25th centile. Height rise up normal values slowly without any differences between children below and over 25th centile. At the age of two years 57 children (83%) became tolerant to cow's milk proteins and after 5-year follow-up in 3 children (4.4%) only persisted cow's milk allergy. All these children presented the IgE-mediate clinical form.

Evidence for genomic changes in transgenic rice (Oryza sativa L.) recovered from protoplasts.

The occurrence of genomic modifications in transgenic rice plants recovered from protoplasts and their transmission to the self-pollination progeny has been verified with the random amplified polymorphic DNA (RAPD) approach. The plant was the Indica-type rice (Oryza sativa L.) cultivar Chinsurah Boro II. The analysed material was: (1) microspore-derived embryogenic rice cells grown in suspension culture, (2) transgenic plants recovered from protoplasts produced from the cultured cells and (3) the self-pollination progeny (two successive generations) of the transgenic plants. DNA purified from samples of these materials was PCR-amplified with different random oligonucleotide primers and the amplification products were analysed by agarose gel electrophoresis. Band polymorphism was scored and used in band-sharing analyses to produce a similarity matrix. Relationships among the analysed genomes were expressed in a dendrogram. The extensive DNA changes evidenced in cultured cells demonstrate the occurrence of somaclonal variation in the material used to produce protoplasts for gene transfer. Quantitatively reduced DNA changes were also found in the resulting transgenic plants and in their self-pollination progenies. While confirming the stability of the foreign gene in transgenic plants, this work gives molecular evidence for the occurrence of stable genomic changes in transgenic plants and points to in vitro cell culture as the causative agent. RAPDs are shown to be a convenient tool to detect and estimate the phenomenon at the molecular level. The methodology is also proposed as a fast tool to select those transgenic individuals that retain the most balanced genomic structure and to control the result of back-crosses planned to restore the original genome.

[Evaluation of sensitivity and specificity of antigliadin antibodies for the diagnosis of celiac disease in childhood]. / Valutazione della sensibilità e della specificità degli anticorpi antigliadina per la diagnosi di malattia celiaca del bambino.

A retrospective study with the aim of evaluating sensitivity and specificity of antigliadin antibodies (AGA) was carried out. AGA values, IgA and IgG, and mucosal damage at intestinal biopsy were compared in each subject, on a sample of 245 subjects, who had undergone intestinal biopsy because of suspected coeliac disease (CD), from January 1991 to December 1993. 130 subjects (53.1%) were found to be suffering from CD, whereas the remaining 115 subjects (46.9%), who did not show any signs of the disease, were considered as controls. When we compared AGA values with mucosal damage we observed that IgA were normal in 24 (18.5%) and pathological in 106 (81.5%) coeliac patients, whereas they were normal in 107 (93%) and pathological in 8 (7%) controls. IgG values were normal in 17 (13.1%) and pathological in 113 (86.9%) coeliac patients, but normal in 75 (65.2%) and pathological in 40 (34.8%) controls. Data about AGA sensitivity related to age, showed IgA sensitivity varying from 80.4% in children under 1 year old to 70% in children over 10 years old and IgG sensitivity from 94.1% to 50%. Data about specificity related to age, showed IgA sensitivity varying from 78.7% in children under 1 year old to 100% in children over 10 years old and IgG specificity from 30.4% to 90.9%. Our results indicated that AGA-IgA had a total sensitivity or 81.5%, a total specificity of 93% and a concordance with intestinal damage of 86.9% while AGA-IgG had a total sensitivity of 86.9%, a total specificity of 65.2% and a concordance with intestinal damage of 76.7%. These data presented a modification on the basis of age, being higher in children under 3 years of age and decreasing with the increase in age. We conclude that AGA have great importance in suspect CD, but it is most important that their results are evaluated on the basis of age.

[Invasive cardiological diagnosis in an ambulatory regimen of transported inpatients]. / Diagnostica cardiologica invasiva in regime ambulatoriale protette.

We report our initial experience with 302 consecutive cases of percutaneous cardiac catheterization in in-patients from other hospitals. The patients reached our laboratory immediately before the procedure in an ambulance with an attending physician and were transported back to their hospital soon after completion of the procedure. This accounts for 35% of 864 diagnostic cardiac catheterization procedures in our laboratory in the first 12 months of activity. There were no complications related to this regimen, and a substantial reduction in unnecessary overnight admission to the cardiology ward was achieved. This report confirms the safety and the advantages of this practice. Implications for the organization of the catheterization laboratory are discussed.

Hybrid rice (Oryza sativa L.): identification and parentage determination by RAPD fingerprinting.

DNA from three families of rice plants selected in Northern China (each comprising the male sterile, the restorer, the hybrid F1 and the maintainer lines) has been extracted and amplified by PCR with different random DNA primers (RAPD analysis). Then, DNA has been analysed by agarose gel electrophoresis and DNA bands scored as present or absent. The generated matrices are reproducible and amenable for identification of each single plant line. Thus, RAPD fingerprinting of the inbred parental lines and of the resulting hybrid is proposed as a convenient tool for the identification, protection and parentage determination of plant hybrids. Furthermore, by offering a molecular tool to verify the degree of dissimilarity between the parental lines, the RAPD analysis may also be used to search for new parental combinations.

Non-ionophoretic elevation of intracellular Ca2+ by Lonidamine.

Lonidamine is an antispermatogenic and anticancer drug that is believed to act by inhibition of energy metabolism. In this study, the effects of Lonidamine on the concentration of intracellular free Ca2+ of several tumor cell lines were assessed because of the important role that cytosolic Ca2+ plays in cell viability and proliferation. The presence of 300 microM Lonidamine resulted in large elevations of cytosolic Ca2+ (> 100 nM) in AS-30D rat ascites hepatoma cells and in cultured EMT6 murine mammary adenocarcinoma cells but had little effect on cultured NCI-H345 human small cell lung cancer cells. The apparent EC50 for Lonidamine was approximately 175 microM. The source of elevated cytosolic Ca2+ was primarily intracellular stores, and the effects of Lonidamine on Ca2+ efflux from these stores did not appear to be due to an ionophoretic action of this compound or to a decline in the level of cellular ATP. These results indicate that the Ca2+ homeostasis of certain lines of tumor cells is specifically altered by Lonidamine at concentrations known to affect cell proliferation.

Rhein inhibits glucose uptake in Ehrlich ascites tumor cells by alteration of membrane-associated functions.

Rhein (RH), 4,5 dihydroxyanthraquinone-2-carboxylic acid, is known to inhibit the glycolysis of neoplastic cells by impairing glucose uptake. In order to establish whether this might be due to a selective interaction of the carrier with the drug or to functional modifications of the cell membrane, the effect of RH on glucose uptake in Ehrlich ascites tumor cells has been investigated. RH strongly inhibits the uptake of both 2-deoxyglucose and 3-O-methylglucose, so the reduced influx therefore cannot be ascribed to an effect on glucose phosphorylation. The inhibition of glucose transport does not depend on a reduction of the number of the carriers as indicated by the inability of the drug to interfere with the synthesis of the transporter. Moreover, the extent of total binding of cytochalasin B, as well as the fact that glucose specificity is not altered, indicate that the intrinsic activity of the glucose carrier is not affected. We therefore conclude that the inhibition of glucose uptake must be ascribed to an interaction of the drug with cell membranes that results in an alteration of membrane-associated functions.

State of the foreign gene and of the genome in transgenic rice (Oryza sativa L.).

PCR with random primers (RAPD analysis) performed on the DNA of embryogenic and non-embryogenic suspension cultured rice and of transformed rice plants allows the evaluation of the extent of DNA changes in the different biological materials. This is thus suggested as a convenient approach, in combination with restriction analysis and Southerns blotting, to evaluate the integrity of the foreign gene, the stability of the insertion site and the stability of the whole genome.

Thermal behavior of a human glioma cell line and its response to combinations of hyperthermia and lonidamine.

The effect of hyperthermia and lonidamine, alone and in combination, on the clonogenic activity of a human glioma cell line was investigated. The time-temperature relationship of asynchronous, exponentially growing cells was defined in the range of 40-45 degrees C. All survival curves were exponential and an Arrhenius plot for heat killing was linear over the temperature range tested, with an activation energy of 192 Kcal/mol. The survival curve of lonidamine-treated cells was also exponential after an initial shoulder. The analysis of the interaction between lonidamine and hyperthermia, performed by the isobolar method, demonstrated an additivity of response so that the effectiveness of the combined treatment was the result of two independent effects. Lonidamine inhibits the neoplastic growth mainly through an ATP depletion, but the thermal killing was not mediated by the drug-induced changes in the energy status of the cell. The effectiveness of the combined treatment was strongly influenced by the schedule of administration. In fact, the sequence lonidamine-->hyperthermia made the cells less sensitive to heat so that the pre-established end-point, i.e. 30% survival, was never achieved whichever combination was used. This "drug-induced heat resistance" was not associated with the induction of heat shock proteins, but rather with modification of cell cycle. On the contrary, showing a purely additive effect, the sequence hyperthermia-->lonidamine allowed achievement of the pre-established cell killing (70%), with exposure times (1-2 hr) and with a temperature (42 degrees C) generally accepted as clinically achievable. Therefore, also considering its low systemic toxicity, lonidamine may be useful in reducing the side effects of hyperthermia.