Characterization of MADS-box genes in charophycean green algae and its implication for the evolution of MADS-box genes

The MADS-box genes of land plants are extensively diverged to form a superfamily and are important in various aspects of development including the specification of floral organs as homeotic selector genes. The closest relatives of land plants are the freshwater green algae charophyceans. To study the origin and evolution of land plant MADS-box genes, we characterized these genes in three charophycean green algae: the stonewort Chara globularis, the coleochaete Coleochaete scutata, and the desmid Closterium peracerosum-strigosum-littorale complex. Phylogenetic analyses suggested that MADS-box genes diverged extensively in the land plant lineage after the separation of charophyceans from land plants. The stonewort C. globularis mRNA was specifically detected in the oogonium and antheridium together with the egg and spermatozoid during their differentiation. The expression of the C. peracerosum-strigosum-littorale-complex gene increased when vegetative cells began to differentiate into gametangial cells and decreased after fertilization. These expression patterns suggest that the precursors of land plant MADS-box genes originally functioned in haploid reproductive cell differentiation and that the haploid MADS-box genes were recruited into a diploid generation during the evolution of land plants.     

More genes underwent positive selection in chimpanzee evolution than in human evolution

Observations of numerous dramatic and presumably adaptive phenotypic modifications during human evolution prompt the common belief that more genes have undergone positive Darwinian selection in the human lineage than in the chimpanzee lineage since their evolutionary divergence 6-7 million years ago. Here, we test this hypothesis by analyzing nearly 14,000 genes of humans and chimps. To ensure an accurate and unbiased comparison, we select a proper outgroup, avoid sequencing errors, and verify statistical methods. Our results show that the number of positively selected genes is substantially smaller in humans than in chimps, despite a generally higher nonsynonymous substitution rate in humans. These observations are explainable by the reduced efficacy of natural selection in humans because of their smaller long-term effective population size but refute the anthropocentric view that a grand enhancement in Darwinian selection underlies human origins. Although human and chimp positively selected genes have different molecular functions and participate in different biological processes, the differences do not ostensibly correspond to the widely assumed adaptations of these species, suggesting how little is currently known about which traits have been under positive selection. Our analysis of the identified positively selected genes lends support to the association between human Mendelian diseases and past adaptations but provides no evidence for either the chromosomal speciation hypothesis or the widespread brain-gene acceleration hypothesis of human origins.     

Type I and type II keratins have evolved from lower eukaryotes to form the epidermal intermediate filaments in mammalian skin.

We have traced the evolutionary origins of keratin-like sequences to the genomes of lower eukaryotes. The proteins encoded by these genes have evolved to form the intermediate filaments that comprise the backbone of vertebrate skin cells. Two related but distinct types of keratins encoded by two separate multigene subfamilies are expressed in the epidermal keratinocytes of vertebrate species from fish to human. Both at the level of protein and at the level of DNA, these two classes of keratins are coordinately conserved throughout vertebrate evolution, indicating the central role that both types of keratins must play in the assembly and structure of the 8-nm filament.     

Type I and type II T cell profiles in chronic myelogenous leukemia

T cell immunity is considered to play an important role in the control of cell growth in chronic myelogenous leukemia (CML) although information regarding the characteristics of T lymphocytes in CML patients is limited. Using flow cytometric analysis of intracellular cytokine expression at the single-cell level, we analyzed helper T and cytotoxic T subsets in 8 CML patients. The percentage of interferon-gamma (IFN-gamma) single-positive CD4 cells (Th1) and that of interleukin-4 (IL-4) single-positive CD4 cells (Th2) was markedly decreased in pretreated CML patients compared to normal controls. In addition, the percentage of IFN-gamma/IL-4 double-positive CD4 cells (Th0) was also reduced. Consequently, the percentage of IFN-gamma/IL-4 double-negative CD4 cells was markedly increased. Similarly, a reduction in IFN-gamma single-positive CD8 cells (Tc1) and IFN-gamma/IL-4 double-positive CD8 cells (Tc0) and an increase in IFN-gamma/IL-4 double-negative CD8 cells were observed in pretreated CML patients. Imbalance of these parameters was markedly improved following cytoreduction therapy. Our findings directly indicate anergic states in CML patients. Determination of the factors that affect Th and Tc profiles may lead to further understanding of immunological states and the development of effective immunotherapy.     

The costs of the detrimental effects of alcohol abuse have grown faster than alcohol consumption in Finland

Between 1980 and 1990 alcohol consumption in Finland grew on average by 2.4% per year, and most of the detrimental effects of alcohol abuse grew faster. The real costs from nearly all alcohol-related detrimental effects grew during the 1980s on average by 1.7–2.4% annually, depending on the item. As a result of the volume and cost development the direct detrimental effects of alcohol abuse grew from FIM 1.0–1.3 billion in 1980 to FIM 2.8–3.7 billion in 1990: i.e. a real increase of 51–56% in the direct costs of detrimental effects. The indirect costs of detrimental effects (production losses, value of life lost through premature death), was FIM 9.9–18.1 billion in 1990. In 10 years the distribution of the costs of direct detrimental effects changed markedly, in particular regarding health and social costs: the share of health costs decreased 6 percentage points, while that of social costs increased 10 percentage points.     

Type I and type II T-cell profiles in aplastic anemia and refractory anemia

Immune-mediated suppression of hematopoiesis has been considered the most important mechanism in the development of idiopathic aplastic anemia (AA) and some types of myelodysplastic syndrome (MDS). Using flow cytometric analysis of intracellular cytokine expression at the single-cell level, we analyzed helper-T (Th) and cytotoxic-T (Tc) subsets in patients with AA and MDS of refractory anemia (RA) subtype. The results showed marked increases in the Th1/Th2 ratio and Tc1/Tc2 ratio in both AA and MDS/RA. Overproduction of cytokines inhibitory to hematopoiesis reported so far might be a reflection of imbalanced development of Th and Tc. Determination of the factors that affect Th and Tc profiles may lead to further understanding of immunological states and the development of effective immunotherapy in AA as well as some types of MDS.     

Hypertensive type I diabetics have a higher level of microalbuminuria than normotensives irrespective of blood glucose equilibrium

Micro-albuminuria, glycosylated haemoglobin and creatinine clearance were measured in 28 insulin dependent and 8 insulin requiring diabetic patients (17 males aged 45 +/- 14.4 and 19 females aged 40 +/- 17 years). Twelve had retinopathy and six had high blood pressure. There was no statistical difference of duration of diabetes, creatinine clearance and glycosylated haemoglobin between patients with normal microalbuminuria and patients with hyper microalbuminuria. There was no correlation between glycosylated haemoglobin and micro-albuminuria, between creatinine clearance and micro-albuminuria, and between creatinine clearance and glycosylated haemoglobin. Patients with retinopathy did not have significantly higher levels of micro-albuminuria. Hypertensive patients had a significantly higher micro-albuminuria (p = 0.03, m = 45.10 +/- 56) compared with normotensive patients (m = 15.49 +/- 17.54). Glycosylated haemoglobin did not differ whether patients were normo or hypertensive. Hypertensive patients were significantly older (p = 0.01) than normotensive. The lack of difference in glycemic control between normo and hypertensive diabetics suggests that diabetes and hypertension are two independent cumulative risk factors of micro-albuminuria. Hypermicro-albuminuria may be secondary to microangiopathy in young diabetics and secondary to hypertension in old diabetics.     

Women with pathologic stage I, II, and III non-small cell lung cancer have better survival than men

OBJECTIVE: Bronchogenic malignancy is the number one cause of cancer deaths in both men and women worldwide. National registry-based studies have shown gender disparity in clinicopathologic characteristics and in survival. This study evaluates the risk factors and trends of lung cancer between genders. METHODS: A prospective cohort of consecutive patients with non-small cell lung cancer (NSCLC) who were carefully clinically (all underwent dedicated positron emission tomography scans) and pathologically staged with stage I, II, or III disease underwent homogenous treatment algorithms and were followed up over a period of 7 years. Primary outcomes were 5-year survival and response to neoadjuvant therapy. RESULTS: There were 1,085 patients (671 men and 414 women). Groups were similar for race, pulmonary function, smoking history, comorbidities, neoadjuvant therapy, histology, and resection rates. Women were younger (p = 0.014), had a higher incidence of adenocarcinoma (p = 0.01), and presented at an earlier pathologic stage (p = 0.01) than men. The overall age-adjusted and stage-adjusted 5-year survival rate favored women (60% vs 50%, respectively; p < 0.001). Women had better stage-specific 5-year survival rates (stage I disease, 69% vs 64%, respectively [p = 0.034]; stage II disease, 60% vs 50%, respectively [p = 0.042]; and stage III disease, 46% vs 37%, respectively [p = 0.024]). Women who received neoadjuvant chemotherapy alone (n = 76) were more likely to be a complete or partial responder than men (n = 142; p = 0.025). CONCLUSIONS: Despite uniform staging and treatment, the 5-year survival rate of women with stage I to III NSCLC was better than men overall and at each stage. Women are more likely to have adenocarcinoma, to present with earlier stage disease, and to be younger. Interestingly, women respond better to neoadjuvant chemotherapy.     

Type I and type II insulin-like growth factor receptors and their function in human Ewing's sarcoma cells

Summary Binding studies using recombinant human¹²⁵I-labelled insulin-like growth factor I ([¹²⁵I]IGF-I) revealed IGF-I receptors in three Ewing's sarcoma cell lines withK _d ranging from 74×10^–12 M to 100×10^–12 M andB _max=36–63 fmol/mg cell protein. [¹²⁵I]IGF-I binding was displaced by IGF-I, IGF-II and insulin with IC₅₀ values of 1.5 nM, 6.3 nM and 0.7 M respectively. Recombinant human [¹²⁵I]IGF-II radioligand-binding assays in the cell lines disclosed specific binding sites for IGF-II withK _d=(110–175)×10^–12 M andB _max varying from 21 fmol/mg to 72 fmol/mg cell protein. Neither IGF-I nor insulin displaced [¹²⁵I]IGF-II binding. IGF-I was found to increase basal glucose transport by maximally 1.5 times with EC₅₀=0.9 nM IGF-I. The efficacy and potency of IGF-II on glucose uptake were comparable to those of IGF-I whereas insulin was ineffective. IGF-I and IGF-II also provoked stimulation of glycogen synthesis in Ewing's sarcoma cells. The maximal glycogenic response was reached at 0.01 M IGF-I and 0.1 M IGF-II, the EC₅₀ value being approximately 1 nM IGF-I and 2 nM IGF-II. Insulin did not significantly influence glycogen formation. IGF-I and IGF-II but not insulin increased DNA synthesis in Ewing's sarcoma cells. The maximal mitogenic response was obtained with 10 nM IGF-I or IGF-II with an EC₅₀ value of about 0.7 nM for both peptides. -IR-3, a monoclonal antibody specific for the IGF type I receptor, effectively blocked IGF-I- and IGF-II-mediated metabolic responses. In conclusion, the data show that IGF-I and IGF-II induce rapid and longterm biological responses in Ewing's sarcoma cells exclusively through interaction with IGF type I receptors.     

The Antennapedia-type homeobox genes have evolved from three precursors separated early in metazoan evolution.

The developmental control genes containing an Antennapedia-type homeobox are clustered in insects and vertebrates. The evolution of these genes was studied by the construction of evolutionary trees and by statistical geometry in sequence space. The comparative analysis of the homeobox sequences reveals the subdivision of the Antennapedia-type homeobox genes into three classes early in metazoan evolution. This observation suggests an important function of these genes even in the most primitive metazoans. Subsequent duplication events generated a cluster of at least five homeobox genes in the last common ancestor of insects and vertebrates. These genes later independently gave rise to the 13 groups of paralogous genes in vertebrates and to the 11 Antennapedia-type genes in the Drosophila complexes.     

Serum and erythrocyte magnesium levels in type I and type II diabetics

Summary Serum and red blood cell magnesium (RBC-Mg) concentrations of 195 type I and III type II diabetic outpatients with different degree of control and 40 control subjects have been evaluated using atomic absorption spectrophotometry. In the total group, no significant difference in serum and RBC-Mg levels in type I and II diabetic outpatients could be found. However, poor control was often associated with lower serum magnesium levels. A negative correlation was found between serum magnesium levels and HbA₁. A particular group of male patients with severe macroangiopathy showed high RBC-Mg levels; this finding was probably due to atherosclerotic and hypertensive renal involvement.     

Physical training decreases plasma thrombomodulin in Type I and Type II diabetic patients

Aims/hypothesis. Endothelial damage is an early step in the pathogenesis of atherosclerosis and its improvement through physical training can contribute to the known reduction of cardiovascular risk associated with exercise. An increase in some endothelium-dependent haemostatic parameters, considered as markers of endothelial damage, has been observed in diabetic patients. Methods. The effect of a three-month physical exercise programme on thrombomodulin, tissue factor pathway inhibitor, plasminogen activator inhibitor, tissue-type plasminogen activator and von-Willebrand factor was evaluated in 14 well-controlled patients with Type I (insulin-dependent) diabetes mellitus and 13 patients with Type II (non-insulin-dependent) diabetes mellitus (HbA_{1 c} 6.5 ± 0.8 and 7.4 ± 0.8 %, respectively). A matched control group was also studied. Results. Thrombomodulin at baseline was higher in both Type I and Type II diabetic patients than in their respective matched control subjects (50.0 ± 16 vs 31.1 ± 8.7 μg/l, p < 0.05; 51.0 ± 10 vs 28.5 ± 11 μg/l, p < 0.05, respectively). After the exercise programme, thrombomodulin plasma concentrations had decreased (p < 0.05) in both groups of patients, with final thrombomodulin values being similar to those observed in their control groups (38.2 ± 11 μg/l for Type I and 34.6 ± 12 μg/l for Type II patients). The thrombomodulin decrement correlated with baseline thrombomodulin and VO_2max increase in Type I diabetic patients. A decrease in tissue factor pathway inhibitor was also observed in Type II diabetic patients. Conclusion/interpretation. We conclude that the normalisation of plasma thrombomodulin concentrations in Type I and Type II diabetic patients after physical training might reflect the improvement in endothelial function associated with physical exercise. [Diabetologia (2001) 44: 693–699]     

Aldosterone-stimulated down-regulation of both type I and type II adrenocorticosteroid receptors in mouse brain is mediated via type I receptors

The concentrations of type I and type II adrenocorticosteroid receptors in brain cytosol obtained from adrenalectomized-ovariectomized female mice were measured with five different assay conditions. Among the five brain regions studied, hippocampus had the highest concentration of type I receptors, whereas cerebral cortex had the highest concentration of type II receptors. The value of properly correcting for dexamethasone cross-binding to type I receptors when type II receptors are being assayed was demonstrated using the type II receptor-selective ligand RU28362. A time-course study revealed a transient up-regulation of both receptor classes in most brain regions after adrenalectomy-ovariectomy, with maximal values achieved 3-5 days postsurgery and a reduction to near-intact levels by 16 days postsurgery. A single sc injection of aldosterone given to adrenalectomized-ovariectomized mice produced a profound down-regulation of type I receptors in hippocampal, cerebral cortex, hypothalamic, brain stem, and cerebellar samples, whereas it down-regulated type II receptors only in hippocampal and cerebral cortical samples. A similar injection of RU28362 failed to down-regulate type I receptors in any brain region, but it did reduce the concentration of type II receptors in all brain regions except cerebellum. The actions of aldosterone appear to be mediated solely through type I receptors, since injections of the type I receptor antagonist RU26752 prevented aldosterone-induced down-regulation of both type I and type II receptors, whereas RU26752 had no effect on the down-regulatory actions of RU28362. The ability of aldosterone to down-regulate type I, but not type II, receptors in hypothalamic, brain stem, and cerebellar samples suggests that type I and type II receptors are concentrated in separate populations of cells in these brain regions, whereas in hippocampus and cerebral cortex there is a sufficient degree of colocalization to permit type II receptor down-regulation via the action of aldosterone-type I receptor complexes. We speculate that this action is mediated at least in part at the genomic level by the suppression of type I and type II receptor mRNA synthesis brought about by the interactions of transformed aldosterone-type I receptor complexes with the DNA regulatory elements upstream from the genes for these receptors.