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PubMed2000

Molecular structure and evolution of DNA sequences located at the alpha satellite boundary of chromosome 20.

We have isolated and characterised one PAC clone (dJ233C1) containing a linkage between alphoid and non-alphoid DNA. The non-alphoid DNA was found to map at the pericentromeric region of chromosome 20, both on p and q sides, and to contain homologies with one contig (ctg176, Sanger Centre), also located in the same chromosome region. At variance with the chromosome specificity shown by the majority of non-alphoid DNA, a subset of alphoid repeats derived from the PAC yielded FISH hybridisation signals located at the centromeric region of several human chromosomes, belonging to three different suprachromosomal families. The evolutionary conservation of this boundary region was investigated by comparative FISH experiments on chromosomes from great apes. The non-alphoid DNA was found to have undergone events of expansion and transposition to different pericentromeric regions of great apes chromosomes. Alphoid sequences revealed a very wide distribution of FISH signals in the great apes. The pattern was substantially discordant with the data available in the literature, which is essentially derived from the central alphoid subset. These results add further support to the emerging opinion that the pericentromeric regions are high plastics, and that the alpha satellite junctions do not share the evolutionary history with the main subsets.

Animals↗
PubMed1999

Recent amplification of the human FRG1 gene during primate evolution.

There is evidence of multiple copies of the FSHD Region Candidate Gene 1 (FRG1) in humans. Analysis of human FRG1 ESTs showed many of them to be non-processed pseudogenes dispersed throughout the genome. To determine when the amplification of FRG1 occurred, we used a PCR-based approach to identify FRG1 sequences from great apes, chimpanzee, gorilla and orang-utan, and an Old World monkey, Macaca mulatta. In common with humans, multiple copies of FRG1 were detected in the great apes. However, in Macaca mulatta, only two FRG1 loci were identified, one presumed to be the homologue of the human chromosome 4q gene. This is strikingly similar to the distribution of a dispersed 3.3-kb repeat family in primates. A member of this family, D4Z4, maps to the subtelomeric region of 4q, in close proximity to FRG1. We propose that an ancestral duplication of distal 4q included FRG1. This duplication is present in Macaca mulatta whose divergence from hominoids is thought to have occurred at least 33 million years ago. We propose that this telomeric region then underwent further amplification and dispersion events in the great ape lineage, with copies of FRG1 and the 3.3-kb repeats being localized in heterochromatic regions.

Animals↗
PubMed1997

Quantitative colorimetric and gas chromatographic determination of arecaidine propargyl ester.

Arecaidine propargyl ester (APE) is a potent muscarinic agonist often used in pharmacological studies. To date, no sensitive quantitative analytical method for APE has been published. In this study, two methods for the quantitative determination of APE are compared: a colorimetric assay, based on the formation of the corresponding ferric(III)-hydroxamic acid complex, and a direct gas chromatographic method, using arecoline as the internal standard. The latter method was found to be more precise. The utility of the gas chromatographic assay was further demonstrated in a stability study of the drug in the biological fluid aqueous humor of rabbits.

Animals↗
PubMed1999

Effect of ED management on ICU use in acute pulmonary edema.

Acute pulmonary edema (APE) is a common Emergency Department (ED) presentation requiring admission to an intensive care unit (ICU). This study was undertaken to examine the effect of ED management on the need for ICU admission in patients with APE. ED records of APE patients were abstracted for patient age, prehospital and ED pharmacological treatment, diagnoses, airway interventions, and ICU length of stay (LOS). Statistical analysis was through multiple regression, logistic regression, chi-square, and ANOVA. One hundred eighty-one patients composed the study group. Pharmacological treatment included nitroglycerin (NTG), 147 patients (81%); morphine sulfate (MS), 88 (49%); loop diuretics (LD), 133 (73%); and captopril sublingual (CSL), 47 (26%). Use of CSL and MS were associated with opposing needs for ICU admission. MS use was associated with increased ICU admissions (odds ratio, 3.08; P = .002), whereas CSL use was associated with decreased ICU admissions (odds ratio, 0.29; P = .002). Morphine sulfate use also demonstrated an increased need for endotracheal intubation (ETI) (odds ratio, 5.04; P = .001), whereas CSL demonstrated a decreased need for ETI (odds ratio, 0.16; P = .008). Ninety-three patients required some form of respiratory support. Forty received noninvasive pressure support ventilation (NPSV) from a bilevel positive airway pressure system (BiPAP), and 60 received endotracheal intubation. Some patients received more than 1 form of respiratory support; all other patients received supplemental oxygen only. The ICU-LOS associated with different airway interventions were supplemental oxygen, 0.72 days; BiPAP, 1.48 days; and ETI, 3.70 days (P < .001). Specific ED pharmacological interventions are associated with a decreased need for ICU admission and endotracheal intubation in acute pulmonary edema patients, whereas use of noninvasive pressure support ventilation correlates with a reduction in the ICU length of stay for patients who do require critical care admission.

Acute Disease↗
PubMed1997

Expression of yeast but not human apurinic/apyrimidinic endonuclease renders Chinese hamster cells more resistant to DNA damaging agents.

Abasic sites represent ubiquitous DNA lesions that arise spontaneously or are induced by DNA-damaging agents. They block DNA replication and are considered to be cytotoxic and mutagenic. The key enzymes involved in the repair of abasic sites are apurinic/apyrimidinic (AP) endonucleases which process these lesions in an error-free mechanism. To analyze the role of AP endonuclease in the protection of mammalian cells against DNA damaging agents, we have transfected both the human (APE) and the yeast (APN1) AP endonuclease in Chinese hamster cells and compared the effects of expression of these genes in stable transfectants as to survival of cells and formation of chromosomal aberrations. Although APE was markedly expressed on RNA and protein level, nuclear extracts of human APE transfectants did not show a higher AP endonuclease activity than the parental line and became not more resistant to the cell killing and clastogenic effect of methyl methanesulfonate (MMS) and hydrogen peroxide (H2O2). In contrast, cells transfected with the yeast APN1 gene expressed higher AP endonuclease activity and became clearly more resistant to the cytotoxic and chromosome breakage inducing activity of the agents. The results indicate that the excision repair capacity and correspondingly the mutagen resistance can be elevated by introducing, in mammalian cells, a yeast DNA repair gene and verify that AP sites are both cytotoxic and clastogenic lesions.

Animals↗
PubMed2001

Alkylphenolic compounds in edible molluscs of the Adriatic Sea (Italy).

This paper reports the first group of results on alkylphenol (APE) contamination of seafood in the Adriatic Sea, in the framework of a national project on the quality of this Sea (PRISMA 2). Nonylphenol (NP), octylphenol (OP), and their ethoxylates (NPE and OPE) were detected in edible molluscs, either filter feeders or predators (clams, mussels, cuttlefishes, and squids), caught from 15 harbors along the Italian coast in the Adriatic Sea in 1997. NP was the compound found always at levels much higher than the other APEs in all the examined species. It reached the maximum concentration of 696 ng/g fresh weight in the squids from the central Adriatic Sea. OP generally occurred at levels 30 times lower than NP. OP was found up to a level of 18.6 ng/g in squids from central Adriatic Sea. OPE was the compound always spotted at the lowest concentrations, up to 0.43 ng/g. NPE was always below the detection limit. The pattern of contamination in the three areas examined was different between bivalve and cephalopod species. No exhaustive risk assessment for marine organisms and human health can be conducted on the basis of these results because data are insufficient. Yet, the occurrence of NP suggests a negligible risk for mussels, which represent the only molluscs for which data are adequate. As to the possible human health implications, the consumption of molluscs of the Adriatic Sea implies APE intakes that are some orders of magnitude lower than those responsible for toxic effects in laboratory animals. Despite these apparently low risks for mussels and human health, the reasons for concern still remain because the levels of alkylphenols found in this study indicate a general contamination of the Adriatic Sea even far from the cost. Furthermore, these levels might represent an unacceptable hazard for other marine organisms. Finally, they contribute to the general environmental estrogen pool.

Animals↗
PubMed1999

Localization of subtelomeric sequences of human chromosomes 1q, 11p, 13q, and 16q in the higher primates.

Relative phylogenetic divergence of the members of the Pongidae family has been based on genetic evidence. The recent isolation of subtelomeric probes specific for human (HSA) chromosomes 1q, 11p, 13q, and 16q has prompted us to cross-hybridize these to the chromosomes of the chimpanzee (Pan troglodytes, PTR), gorilla (Gorilla gorilla, GGO), and orangutan (Pongo pygmaeus, PPY) to search for their equivalent locations in the great apes. Hybridization signals to the 1q subtelomeric DNA sequence probe were observed at the termini of human (HSA) 1q, PTR 1q, GGO 1q, PPY 1q, while the fluorescent signals to the 11p subtelomeric DNA sequence probe were observed at the termini of HSA 11p, PTR 9p, GGO 9p, and PPY 8p. Fluorescent signals to the 13q subtelomeric DNA sequence probe were observed at the termini of HSA 13q, PTR 14q, GGO 14q, and PPY 14q, and positive signals to the 16p subtelomeric DNA sequence probe were observed at the termini of HSA 16q, PTR 18q, GGO 17q, and PPY 19q. These findings apparently suggest sequence homology of these DNA families in the ape chromosomes. Obviously, analogous subtelomeric sequences exist in apes' chromosomes that apparently have been conserved through the course of differentiation of the hominoid species.

Animals↗
PubMed2002

Antiphospholipid antibodies and kininogens in pathologic pregnancies: a review.

PROBLEM: Recently, evidence has accumulated for the presence of the kallikrein-kinin system or plasma contact system in the fetoplacental unit. The Kallikrein-kinin system or plasma contact system consists of three essential plasma proteins. These are coagulation factor XII, prekallikrein and high molecular weight kininogen. Deficiencies of these proteins and antiphospholipid antibodies are frequent hemostasis-related abnormalities found in unexplained recurrent aborters. METHOD OF STUDY: Review of existing data. RESULTS: Reports of antiphosphatidylethanolamine antibodies (aPE) with similar or identical pathogenic associations as those described for anticardiolipin antibodies (aCL) are found in the literature. We showed a strong association between recurrent pregnancy losses and aPE, the latter of which recognizes kininogens, and kininogen-binding proteins, factor XI and prekallikrein. The reports of aPE are reviewed, the function of the kininogens are summarized, and their role in pregnancy is discussed. CONCLUSIONS: Because kallikrein-kinin system may play an important role in pregnancy especially in fetoplacental unit, disruption of this system may be a risk factor for early gestational losses.

Abortion, Habitual↗
PubMed2003

Antiphospholipid antibodies in implantation failures.

PROBLEM: The amino phospholipids (PL), phosphatidylserine (PS) and phosphatidylethanolamine (PE) are distributed asymmetrically in the plasma membranes of eucaryotic cells. This arrangement involves active transport of PS and PE from the outer to inner membrane leaflet by an aminophospholipid translocase (flipase). Cell activation, injury and programmed cell death (apoptosis) cause collapse of the PS/PE asymmetry by activation of another enzyme system, scramblase. Unlike other cells, the developing trophoblast exteriorizes PS during its differentiation. METHODS OF STUDY: An analysis of published and unpublished data. RESULTS: The trophoblast is targeted by antiphospholipid antibodies (aPL), especially to PS (aPS). Cardiolipin is not present in the trophoblast plasma membrane, nonetheless, anticardiolipin (aCL) has been implicated in trophoblast pathology. The aPS and aCL are often crossreactive. Both animal and in vitro experimental models have shown monoclonal and polyclonal aPS and aCL to specifically destroy trophoblast, inhibit syncytium formation, halt human chorionic gonadatropin (hCG) production, and limit trophoblast invasion. Antibodies to PE (aPE) have not been well characterized, however, recent reports from several independent laboratories document that aPE are associated significantly with very early (embryonic) recurrent pregnancy loss (RPL). Umeda and coworkers have shown that during cytokinesis (late telophase) of Chinese hamster ovary (CHO) cells, formation of PE rafts in cleavage furrows is required for completion of cell division and formation of daughter cells. This raises the question whether aPE might interfere with implantation and cell division during embryogenesis. CONCLUSIONS: A role for aPL in implantation failure and occult pregnancy loss constitutes the basis of this overview.

Animals↗
PubMed2006

Personality and subjective well-being in orangutans (Pongo pygmaeus and Pongo abelii).

Orangutans (Pongo pygmaeus and Pongo abelii) are semisolitary apes and, among the great apes, the most distantly related to humans. Raters assessed 152 orangutans on 48 personality descriptors; 140 of these orangutans were also rated on a subjective well-being questionnaire. Principal-components analysis yielded 5 reliable personality factors: Extraversion, Dominance, Neuroticism, Agreeableness, and Intellect. The authors found no factor analogous to human Conscientiousness. Among the orangutans rated on all 48 personality descriptors and the subjective well-being questionnaire, Extraversion, Agreeableness, and low Neuroticism were related to subjective well-being. These findings suggest that analogues of human, chimpanzee, and orangutan personality domains existed in a common ape ancestor.

Animals↗
PubMed2001

Mental evolution and development: evidence for secondary representation in children, great ages, and other animals.

Recent interest in the development and evolution of theory of mind has provided a wealth of information about representational skills in both children and animals. According to J. Perner (1991), children begin to entertain secondary representations in the 2nd year of life. This advance manifests in their passing hidden displacement tasks, engaging in pretense and means-ends reasoning, interpreting external representations, displaying mirror self-recognition and empathic behavior, and showing an early understanding of "mind" and imitation. New data show a cluster of mental accomplishments in great apes that is very similar to that observed in 2-year-old humans. It is suggested that it is most parsimonious to assume that this cognitive profile is of homologous origin and that great apes possess secondary representational capacity. Evidence from animals other than apes is scant. This analysis leads to a number of predictions for future research.

Animals↗
PubMed2005

A meta-analysis of primate hand preferences, particularly for reaching.

P. F. MacNeilage, M. G. Studdert-Kennedy, and B. Lindblom (1987) proposed a progression for handedness in primates that was supposed to account for the evolution of a right bias in human handedness. To test this proposal, the authors performed meta-analyses on 62 studies that provided individual data (representing 31 species: 9 prosimians, 6 New World monkeys, 10 Old World monkeys, 2 lesser apes, and 4 greater apes), of the 118 studies of primate handedness published since 1987. Although evidence of a population-level left-handed bias for prosimians and Old World monkeys supports P. F. MacNeilage et al., the data from apes, New World monkeys, and individual species of prosimians and New World monkeys do not. Something other than primate handedness may have been the evolutionary precursor of the right bias in hand-use distribution among hominids.

Animals↗
PubMed1976

Evolution of type C viral genes: evidence for an Asian origin of man.

Old World monkeys and apes, including man, possess, as a normal component of their cellular DNA, gene sequences (virogenes) related to the RNA of a virus isolated from baboons. A comparison of the viral gene sequences and the other cellular sequences distinguishes those Old World monkeys and apes that have evolved in Africa from those that have evolved in Asia. Among the apes, only gorilla and chimpanzee seem by these criteria to be African, whereas gibbon, orang-utan and man are identified as Asian, leading us to conclude that most of man's evolution has occurred outside Africa.

Africa↗
PubMed1996

Biosynthesis of anandamide and related acylethanolamides in mouse J774 macrophages and N18 neuroblastoma cells.

Anandamide (arachidonoylethanolamide, AnNH) has been recently proposed as the endogenous ligand at the brain cannabinoid receptor CB1. Two alternative pathways have been suggested for the biosynthesis of this putative mediator in the central nervous system. Here we present data (1) substantiating further the mechanism by which AnNH is produced by phospholipase D (PLD)-catalysed hydrolysis of N-arachidonoylphosphatidylethanolamine in mouse neuroblastoma N18TG2 cells, and (2) suggesting for the first time that AnNH is biosynthesized via the same mechanism in a non-neuronal cell line, mouse J774 macrophages, together with other acylethanolamides and is possibly involved in the control of the immune/inflammatory response. Lipids from both neuroblastoma cells and J774 macrophages were shown to contain a family of N-acylphosphatidylethanolamines (N-aPEs), including the possible precursor of AnNH, N-arachidonoyl-PE. Treatment with exogenous PLD, but not with exogenous phospholipase A2 and ethanolamine, resulted in the production of a series of acylethanolamides (AEs), including AnNH, from both cell types. The formation of AEs was accompanied by a decrease in the levels of the corresponding N-aPEs. Enzymically active homogenates from either neuroblastoma cells or J774 macrophages were shown to convert synthetic N-[3H]arachidonoyl-PE into [3H]AnNH, thus suggesting that in both cells an enzyme is present which is capable of catalysing the hydrolysis of N-aPE(s) to the corresponding AE(s). Finally, as previously shown in central neurons, on stimulation with ionomycin, J774 macrophages also produced a mixture of AEs including AnNH and palmitoylethanolamide, which has been proposed as the preferential endogenous ligand at the peripheral cannabinoid receptor CB2 and, consequently, as a possible down-modulator of mast cells. On the basis of this as well as previous findings it is now possible to hypothesize for AnNH and palmitoylethanolamide, co-synthesized by macrophages, a role as peripheral mediators with multiple actions on blood cell function.

Animals↗
PubMed2000

Progress in understanding hominoid dental development.

Teeth preserve a record of the way they grow in the form of incremental markings in enamel, dentine and cementum. These make it possible to reconstruct cellular activity and the timing of dental development in living and fossil primates, including hominids. They also provide a way of exploring the mechanisms that underlie morphological change during evolution and the nature of the relationship between ontogeny and phylogeny. All living great apes are dentally mature by about 11 y, irrespective of their body mass. While the early period of root formation in living great apes is shorter than in modern humans, enamel takes approximately the same time to form, irrespective of how thick it is. In general, differences in the total time taken to form enamel seem not to be due to differences in the rate at which enamel and dentine are secreted, but rather to faster or slower rates of differentiation of ameloblasts and odontoblasts and therefore to the number of secretory cells active at any one time during tooth formation. Tooth size, especially height, may influence the sequence of appearance of tooth mineralisation stages. The space available in the jaws may also have an influence on both the timing of tooth bud/crypt appearance and the sequence of gingival emergence. When each of these potential influences on dental development are carefully considered, and incremental markings used to calibrate key events, the developing dentition can provide an estimate of the period of dental maturation in fossil hominoids. However, the influence of body mass on the period of dental development among primates remains unclear. The earliest hominoids, dated at around 18 Mya, may still have had modern monkey-like maturational profiles, and the earliest hominids, dated between 1.8 and 3.7 Mya, modern great ape-like maturational profiles. Exactly when the extended or prolonged modern human-like maturational profile first appeared remains debatable, but the most secure suggestion might be at the time of the appearance of the earliest archaic Homo sapiens, when brain size and body mass were finally both within the ranges known for modern humans. But at present we should not reject the hypothesis that an extended, modern human-like, maturational profile arose more than once during human evolution in parallel with an increase in brain size.

Animals↗
PubMed2000

Muscarinic properties of compounds related to arecaidine propargyl ester.

A series of new analogues of the arecaidine propargyl ester (CAS 35516-99-5), APE, 1a) with alcohols consisting of 4 or 5 carbon atoms were investigated at muscarinic receptor subtypes. The muscarinic activity of the quaternary and tertiary salts of the APE-related compounds were assayed on the isolated guinea-pig ileum (M3 receptor subtype) and guinea-pig left atria (M2 receptor subtype) as well as on rabbit isolated vas deferens (M1 receptor subtype). The structural variations made in the APE molecule, replacing the triple bond in the ester side chain with structures such as double bond, an allene moiety, a single bond, a cyclopropyl group or two triple bonds should alter the selectivity and potency in favour of the M2 subtype. Enhanced, though modest, selectivity for M2 receptors was achieved with the 2-butynyl ester 2a. The other structural variations resulted in a loss of potency, but not necessarily of efficacy.

Animals↗
PubMed2005

Andrographolide isolated from Andrographis paniculata induces cell cycle arrest and mitochondrial-mediated apoptosis in human leukemic HL-60 cells.

The in vitro cytotoxicities of the ethanol extract of Andrographis paniculata (APE) and its main diterpenoid components were evaluated in various cancer cells. APE was found to be significantly growth inhibitory to human acute myeloid leukemic HL-60 cells with an IC (50) value of 14.01 microg/mL after 24 h of treatment. Among the three main diterpenoids in A. paniculata, andrographolide exhibited the highest degree of cytotoxicity followed by deoxyandrographolide while neoandrographolide was the least effective. Laser confocal microscopy and gel electrophoresis studies revealed chromosomal DNA fragmentations suggesting the occurrence of apoptosis. An increase of G (0)/G (1) phase cells from 51.88 % to 78.69 % was noted after andrographolide treatment for 36 h. The G (0)/G (1) phase arrest and apoptosis were associated with disappearance of mitochondrial cytochrome c and increased expression of Bax but decreased expression of Bcl-2 proteins in the inhibited cells. Although the order of all these events has not been determined, it is concluded that APE and andrographolide induce cell cycle arrest and affect an intrinsic mitochondria-dependent pathway of apoptosis by regulating the expression of some pro-apoptotic markers in HL-60 cells.

Andrographis↗
PubMed2000

Highly efficient gene transfer into preterm CD34 hematopoietic progenitor cells.

OBJECTIVE: Retrovirus-mediated gene transfer has been shown to transduce CD34(+) cells from term gestation umbilical cord blood with relatively high efficiency. The purpose of this study was to compare the efficiencies of retrovirus-mediated gene transfer into early (23-28 weeks' gestation) and term (37-41 weeks' gestation) umbilical cord blood CD34(+) hematopoietic progenitor cells. STUDY DESIGN: CD34(+) cells were purified from cyropreserved early (23-28 weeks' gestation) and term (37-40 weeks' gestation) umbilical cord blood specimens with fluorescence-activated cell sorting. The CD34(+) cells were then transduced in virus-containing medium (gibbon ape leukemia virus pseudotype vector LAPSN [PG13]) in wells coated with the recombinant human fibronectin fragment CH-296 and in the presence of multiple hematopoietic growth factors (interleukin 6, stem cell factor, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and megakaryocyte growth and development factor) and protamine sulfate. The LAPSN (PG13) virus-containing medium was changed every 12 hours for 48 hours, after which time colony-forming cells were assayed in soft agar. The gibbon ape leukemia virus pseudotype vector LAPSN (PG13) contains the human placental alkaline phosphatase and neomycin phosphotransferase (neo ) genes. The efficiency of gene transfer was assessed by histochemical staining of colony-forming cells in agar for expression of heat-stable alkaline phosphatase. RESULTS: Gene transfers, as assessed by alkaline phosphatase staining of colony-forming cells (granulocyte-macrophage colony-forming units and erythroid burst-forming units), were similar for CD34(+) hematopoietic progenitor cells from early (58.4% +/- 11.8%) and term (63.2% +/- 12.5%) gestation fetal umbilical cord blood. CONCLUSION: CD34(+) hematopoietic progenitor cells from midgestation fetal blood can be transduced with high efficiency using techniques optimized for postnatal samples with a gibbon ape leukemia virus pseudotype vector. The early fetus may be a preferable target for gene therapy because of the higher number of circulating CD34(+) and CD38(-) cells relative to term cord blood, their greater proliferative capacity, and the rapid expansion of the fetal hematopoietic system that occurs from the second trimester to delivery. Because in vitro studies of gene transfer into hematopoietic progenitor cells and long-term culture-initiation cells have not been predictive of the efficiency of gene transfer into marrow-repopulating cells in vivo, studies that examine clinically applicable approaches to in utero gene therapy in appropriate animal models are still needed.

Alkaline Phosphatase↗