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Efficient initiation of translation at non-AUG triplets in plant cells.

The efficiency of translation initiation at triplets differing at one residue from AUG was tested by transient expression in protoplasts from two different plant species. All possible alternative codons were tested. Some triplets showed significant CAT activity, with CUG (30% of the AUG activity) being most active. Most others had between 5 and 15% of the activity obtained from AUG, whereas UUG and AUC yielded about 2% and the two composed only of purines, AAG and AGG, had no significant activity. Translation initiation from AUC, especially, responded to leader sequences outside the immediate context which did not affect translation initiation from AUG.

Base Sequence↗

Multichannel flash spectroscopy of the reaction centers of wild-type and mutant Rhodobacter sphaeroides: bacteriochlorophyllB-mediated interaction between the carotenoid triplet and the special pair.

Multichannel flash spectroscopy (with microsecond time resolution) has been applied to carotenoid (Car)-containing and Car-less reaction centers (RC) of Rhodobacter sphaeroides with a view to investigate the interaction between the Car and its neighboring pigments at room temperature. Under neutral redox potential conditions, where the primary quinone acceptor (QA) is oxidized, the light-induced spectral changes in the 350-1000 nm region are attributed to the photochemical oxidation of the special pair (denoted here as P870), the generation of P870(+)QA(-), and the attendant electrochromism of adjacent chromophores. A bathochromic shift of <1 nm in the visible absorption region of Car reveals the sensitivity of Car to the P870 photooxidation. Under low redox potential conditions, where QA is reduced, P870 triplets (P870(+)) are formed. The time-resolved triplet-minus-singlet (TmS) spectrum of Car-less RC shows a deep bleaching at 870 nm, which belongs to P870(+), and additional (but smaller) bleaching at 800 nm; the entire spectrum decays at the same rate (with a lifetime of about 50 micros). The bleaching at 800 nm arises from the pigment interaction between P870(+) and the accessory bacteriochlorophylls on A and B branches (BA,B). In Car-containing RC, the TmS spectra of Car are accompanied by two smaller, negative signals--a sharp peak at 809 +/- 2 nm and a broad band at 870 nm--which decay at the same rate as the TmS spectrum of Car (ca 10 micros). The former is ascribed to the perturbation, by Car(+), of the absorption spectrum of BB; the latter, to the TmS spectrum of P870(+), a species that appears to be in approximate thermal equilibrium with Car(+). These assignments are consistent with the absorption-detected magnetic resonance spectra obtained by other workers at low temperatures.

Bacteriochlorophylls↗

[Visible absorption spectra and resonance Raman spectra of n-pi* singlet-triplet transition of p-benzoquinone in CS2].

The visible absorption spectra of p-benzoquinone (PBQ) in CS2 were measured, and a weak absorption band around 507 nm attributed to n-pi* singlet-triplet transition was denonstrated. Using the resonance Raman effect excited in liquid-core optical fiber which can enhance Raman intensity by 6-9 orders of magnitude, the authors obtained the 514.5 nm excited resonance Raman (RR) spectra of PBQ at 1439 cn(-1) in the concentration range from 10(-3) to 10(-6) mol x L(-1). The new characteristic RR band is attributed to the symmetric C==O stretch (v(C==O)) of n-pi* singlet-triplet transition of PBQ. The resonance Raman shift is blue-shifted with decreasing concentration. The results of this paper are helpful for understanding the relationship between the electric structure and the photophysical properties of PBQ, and for obtaining more abundant structural information of molecules.

English Abstract↗

Characterization of mammalian neurofilament triplet proteins. Subunit stoichiometry and morphology of native and reconstituted filaments.

The three major proteins of mammalian neurofilaments of molecular weights 179,000 (NF1), 129,000 (NF2), and 66,500 (NF3) have been purified to homogeneity by multiple anion-exchange and hydroxylapatite absorption chromatography in 8 M urea. Silver staining of polyacrylamide gels of the purified proteins show single bands. In order to gain further insight into the molecular organization of the neurofilament triplet proteins, the molar stoichiometries and morphologies of native and reconstituted filaments and those isolated from developing brain were studied. Denaturing polyacrylamide gel electrophoresis followed by quantitative dye-binding analysis shows that the molar ratio of the three components in neurofilaments isolated from bovine spinal cord myelinated nerve is 4:2:1 (NF3:NF2:NF1). Comparison of the molar ratios of each component in neurofilaments isolated from rat, bovine, and human brain shows a variation in the ratio of each of these polypeptides and raises questions about the physiological uniqueness of the molar composition of the neurofilament triplet. Reconstitution of the three bovine polypeptides into 10-nm filaments was accomplished under conditions in which the NF3 protein was limiting. Reassembly of 10-nm filaments with varying amounts of NF2 and NF1 indicate that the NF3 homopolymer has a limiting capacity to bind NF2 and NF1 and is saturated at a molar ratio of 2:2:1 (NF3:NF2:NF1). Isolation of the neurofilament complex at various stages of rat brain maturation indicates that NF3 and NF2 are integrated into the neurofilament complex as early as embryonic day 17, while NF1 copurifies with these proteins at postnatal day 16, eventually reaching a molar stoichiometry of 2:2:1 in the adult rat. The molecular stoichiometry of the neurofilament proteins, the differential integration of these proteins during brain development, and the variation of the molar composition between mammalian species suggest accessory roles for the NF2 and NF1 proteins in the neurofilament complex.

Amino Acids↗

Temporal firing patterns of single units, pairs and triplets of units in the auditory cortex.

The spontaneous and acoustically driven activities of single units, pairs and triplets of units in the auditory cortex were analyzed. Data were obtained in two sets of experiments from nonbehaving awake cats and from a behaving monkey. The results of the two sets of experiments indicated that neighboring neurons usually fire independently. The weak correlations found between pairs of adjacent neurons were mostly indicative of a common input driving both units. In some cases, signs of synaptic interaction between the neurons were found. When triplets of units were considered, it was found that several independent inputs exist, even within a small group of adjacent neurons. When such small groups of neurons were studied in the behaving monkey, it was found that the temporal firing pattern of single neurons and the interactions between pairs of neurons were in some cases dependent on the behavioral state and on the sensorimotor association.

Animals↗

[Use of triplet state exchange disactivation phenomenon for the study of structure and electron conductivity of proteins].

A method for studying protein structure and estimating its electron conducting properties is proposed. The method is based on the kinetic recording of exchange quenching triplet labels and probes phosphorescence by chromophores or paramagnetic centres. It is shown that different types of exchange interactions (spin exchange, exchange energy transfer) between centres with distance R are described approximately by an equation (I = I0 exp-2R/L) where L changes from 0.7 A (absence of electron coupling--system of type I) to 6.5 A (strong electron coupling--system of type II). I (sec-1) corresponds to exchange energy transfer rate constant or exchange integral in the case of spin exchange. Life-times of excited triplet state eosin-isotiocionate labels connected with the terminal NH2-groups of the following preparations were measured by the method of kinetic phosphorescence decay recording: human oxyhemoglobin, methemoglobin, F- and CN-methemoglobin, metmyoglobin, F- and CN-metmyoglobins. The influence of lysozyme of the nitroxyl spin label bound to His-15 group on the phosphorescence spectrum was investigated. The analysis of our and literature data on the exchange interactions between the centres localized on the protein with known structure (hemoglobin, myoglobin, lysozyme, carboangidrase, bacterial ferredoxin) permit us to conclude that in the examined cases the experimental values correspond to model systems of type I and are different from the dependence in systems of type II by 5--15 order. This allows us to use equation (I) for estimation of the distances between the centres on proteins.

Electron Spin Resonance Spectroscopy↗

[Role of triplet exciplex in electron transport].

The role of triplet exciplex in electron transfer was considered in the photooxidation of chlorophylls "a" and "b" with p-benzoquinone and aromatic nitrous compounds. Relationship between the change of free energy delta F of formation of solvated ion-radical, ion-radical pairs and chlorophyll exciplex and dielectric constant of the medium & was investigated. The conclusions obtained were compared with the experimental data on the intermediate products. It was found that photooxidation of chlorophyll may proceed via the formation of the exciplex in the triplet state, while during pheophytin photooxidation no exciplex is formed and the first stage of electron transfer is the formation of the ion-radical pair.

Chlorophyll↗

[Expansion of triplet repeat and neurodegenerative disorders].

Recent studies on human genetic analysis have revealed that there is hitherto unknown category of genomic abnormality in human genetic diseases; an abnormal expansion of trinucleotide repeat. For example, Huntington's disease is caused by the CAG expansion to more than 40 in IT15 gene Up to the present, we have 7 disorders which are revealed to be apparently caused by the triple repeat expansion: fragile X syndrome (two types), myotonic dystrophy, Huntington's disease, spinocerebellar atrophy type 1, dentatorubropallidoluysian atrophy and bulbo-spinal muscular atrophy. In this symposium, I would like to show first a clear inverse relationship between a length of CAG repeat and the age at onset in each Huntington's disease patient in Japan. Next, we would like to review and discuss on 1) the possible role of normal trinucleotide repeat in human gene, 2) the mechanisms for expansion of triplet repeat and for instability of the repeat length, and 3) the actual mechanism for neuronal cell death related to the expansion of triplet repeat.

Cell Death↗

[Population of triplet-excited states of tryptophanyls and tyrosyls in proteins].

The relative limiting concentrations of triplet excited states of tryptophan and tyrosine residues have been measured by the ESR method. The magnitude of limiting concentration of the tryptophan and tyrosine residues T1-states does not vary to fit their numbers in some proteins. A limitation on the concentration of T1-states is accounted for by the fact that in protein molecules there occurs the radiationless excitation energy transfer of chromophores on residues in the triplet state.

Electron Spin Resonance Spectroscopy↗

Formation of singlet oxygen in the thermal decomposition of 3-hydroxymethyl-3,4,4-trimethyl-1,2-dioxetane, a chemical source of triplet-excited ketones.

Triplet-excited carbonyl species can be generated by photoexcitation of carbonyl compounds or alternatively by thermal decomposition of 1,2-dioxetanes. Such electronically excited species are involved in the oxidative modification of biologically important molecules such as DNA. This study demonstrates that thermal decomposition of 3-hydroxymethyl-3,4,4-trimethyl-1,2-dioxetane (HTMD), which is a chemical source of triplet-excited ketones, is accompanied by infrared photoemission at 1270 nm characteristic for singlet oxygen monomol emission. The intensity of infrared photoemission at 1270 nm was solvent-dependent, in agreement with the reported lifetime of singlet oxygen in the employed solvents. Calibration of monomol emission with the endoperoxide of 1,4-dimethylnaphthalene (DMNO2) showed that HTMD (30 mM) produced 25 microMs of singlet oxygen. Thus, the yield of singlet oxygen production in the thermal decomposition of HTMD in carbon tetrachloride is about 0.1%.

Heating↗

Sequence analysis of the CAG triplet repeats region in the Huntington disease gene (IT15) in several mammalian species.

Huntington's disease (HD) is a progressive neurodegenerative disease linked to abnormally expanded CAG repeats in the first exon of the IT15 gene. Neither HD or other glutamine expansion triplet disease has not yet been described in other species as human. In this study, we sequenced the region containing the CAG repeat expansion region of the corresponding IT15 gene in several mammalian species. In all investigated species, the number of CAG found is equal or inferior to the normal range of human IT15 alleles. The longer uninterrupted CAG stretch was found in the pig and consists of 18 CAG. In the other species, the CAG stretch was shorter and/or interrupted by other triplets. The proline encoding region following the CAG stretch is conserved in the mammalian species studied here and the number of proline is few or not reduced compared to the number of the glutamine codon repeats region.

Amino Acid Sequence↗

A very significant correlation between carcinogenic activity of polycyclic hydrocarbons and certain properties of their transition states in the range of the lowest triplet states of the DNA.

A very significant correlation between carcinogenic power of polycyclic aromatic hydrocarbons and their probability of photoinduced double resonance transitions in the UV-range of about 3 to 3.5 eV is shown. The resonance can be interpreted in terms of the lowest triplet state energy of thymine at 3.25 eV and the energy difference between the triplet states of A--T and G--C base pairs. The correlation has been predicted from a hypothesis which describes cell communication by photon interaction within a cell population.

Carcinogens↗

Delayed delivery of surviving triplet after fetal reduction and spontaneous abortion.

In multiple pregnancies, survival of remaining fetuses after premature death and delivery of one fetus is uncommon. We report a case of a triplet pregnancy that was reduced to twins at the 14th gestational week and then had preterm premature rupture of membranes and intrauterine fetal death of one twin at the 17th gestational week. To save the surviving fetus, delivery of the dead fetus and ligation of the umbilical cord at the cervical level were performed. We also performed McDonald cervical cerclage to keep the placenta of the dead fetus as well as that of the surviving one in the uterine cavity. After a series of aggressive procedures, including immediate administration of tocolytic agents, and antibiotic prophylaxis to prevent infection and preterm labor, the surviving fetus was delivered vaginally 73 days later due to intractable uterine contractions. After a 10-week hospital stay, the infant boy, weighing 2,500 g, was discharged without any sequelae. To our knowledge, this was the longest interval between deliveries in a triplet pregnancy reported in Taiwan. With adequate intensive management, a satisfactory outcome of the fetus and mother in such cases is possible.

Abortion, Spontaneous↗

[Hereditary neurological diseases caused by amplification of triplet repetitions].

NEW TYPE OF MUTATION: Repeated sequences of nucleotide triplets can cause two groups of diseases. GROUP I DISEASES: These diseases result from an expansion of a noncoding portion of a gene which may be repeated more than 1000 times. This group includes several multisystem diseases such as the fragile X syndrome and its variants, Steinert's disease and Friedreich's disease in which nervous system disorders are not always predominant. The molecular mechanism of the cellular disorder is probably related to a nonfunctional abnormal protein. GROUP II DISEASES: Huntington's disease, spinobulbar amyotrophy or Kennedy's disease, dentato-rubo-pallidolusian atrophy and spinocerebellar ataxias 1, 2, 3, 6, and 7 are characterized by local expansion of the coding part of a gene. This low-amplitude expansion always involves the CAG triplet and leads to expression of a protein with an abnormal number of glutamines, producing typical neurodegenerative disease almost exclusively limited to the nervous system. The underlying mechanism of the neuronal suffering is probably the production of an abnormal but functional protein. The causes of this type of mutation remain unclear. PERSPECTIVES: Positive diagnosis is now possible with DNA sequencing. While antenatal diagnosis offers essential information for family genetic counselling there is no perspective of therapeutic propositions for the near future. The problems raised by presymptomatic diagnosis must not be underestimated.

DNA Mutational Analysis↗

Fetal hydantoin syndrome in triplets. A unique experiment of nature.

Birth defects associated with phenytoin and phenobarbital taken during pregnancy include craniofacial and limb deformities. These anomalies were found in various degrees of severity in triplets born to a mother taking phenobarbital and phenytoin for an epileptic disorder, giving us a unique opportunity to recognize different manifestations of the hydantoin syndrome in infants that presumably were exposed to the same level of the drugs. These infants were trizygotic and their clinical manifestations varied widely, suggesting the possibility that genetic background may play a significant role in the susceptibility of the fetus to phenytoin and phenobarbital during pregnancy.

Abnormalities, Drug-Induced↗

Congenital cardiovascular malformations in twins and triplets from a population-based study.

Data from the Baltimore-Washington Infant Study of congenital cardiovascular malformations permitted detailed analysis of congenital cardiovascular malformations in 62 twins and 3 triplets and 2303 singleton cases. A probability sample of controls (n = 2793) included 43 twins. The case prevalence of multiple births was 28 of 1000, compared with a 15 of 1000 prevalence among controls (chi 2 = 5.7). There were more girls among case twins than among case singletons and controls (chi 2 = 9.0). Monozygosity was no more frequent in case twins than in controls. Looping defects occurred in 4 monozygotic twin pairs compared with only 1 dizygotic twin pair. The twinning process itself may be implicated in the development of congenital cardiovascular malformations in some of these infants, especially those with looping defects, but concordance of types of defects in 4 of 65 pairs implicates genetic factors in the determination of some forms of congenital cardiovascular malformations.

Diseases in Twins↗

Triplets and higher-order multiple births. Time trends and infant mortality.

OBJECTIVES: To describe changes in rates of higher-order multiple births (triplets and higher) between 1972 and 1989, to compare infant mortality rates in infants of higher-order multiple births and singletons born from 1983 through 1985, and to compare infant mortality rates among higher-order multiples born from 1983 through 1985 with rates among those born in 1960. RESEARCH DESIGN: Population-based analysis of live births (1972 through 1989) and infant deaths (1960 and 1983 through 1985) in the United States. The rate of higher-order multiple births was calculated per 100,000 live births. DATA SOURCE: Computerized national natality files for 1972 through 1989 and national linked birth/infant death data sets for 1960 and 1983 through 1985 from the National Center for Health Statistics, Centers for Disease Control. POPULATION: Live births to white and black women in the United States. INTERVENTIONS: None. MAIN RESULTS: Between 1972 through 1974 and 1985 through 1989 the rate of higher-order multiple births increased by 113% among infants of white mothers and by 22% among infants of black mothers. In whites the increase was mostly age specific and was not due to the upward shift in the maternal age distribution. The increase was particularly large in white women aged 30 through 34 years (152%) and 35 through 39 years (165%) and in more highly educated mothers. In blacks the modest increase in the rate of higher-order multiple births was mostly due to an upward shift in the maternal age distribution. From 1983 through 1985, mortality of infants of higher-order multiple births was about 15 times that of singletons. This was due almost entirely to the lower birth weight distribution of infants of higher-order multiple births. Their weight-specific mortality compared favorably with that of singletons. At 500 through 999 g, mortality was about the same. In weight categories between 1000 and 1999 g, mortality rates in higher-order multiple births were much lower: weight-specific relative risks ranged from 0.30 to 0.73. Between 1960 and 1983 through 1985 infant mortality in higher-order multiple births declined by about 50%. CONCLUSIONS: It is likely that much of the increase in the incidence of higher-order multiple births is due to the rise in the use of ovulation-inducing drugs for the treatment of infertility. This increase and the decline in mortality risk have created a much greater need for medical and social services for infants of higher-order multiple births and their families.

Black or African American↗

Preterm twins and triplets. A high-risk group for severe respiratory syncytial virus infection.

OBJECTIVE: To assess the impact of multiple births and crowded homes on the severity of respiratory syncytial virus illness in preterm infants with bronchopulmonary dysplasia. RESEARCH DESIGN: Retrospective case-control chart review from a prospective longitudinal respiratory illness study. SETTING: Neonatal High-Risk Follow-Up Clinic (outpatient setting) and tertiary care hospitals (inpatient setting). PARTICIPANTS: Fourteen sets of twins and two sets of triplets followed up between 1983 and 1989 and matched with 34 singleton infants for date of birth (within 3 months) and gestational age (within 1 month). MEASUREMENTS/MAIN RESULTS: The risk of developing respiratory syncytial virus illness was significantly higher in multiple-birth infants than in singletons (53% vs 24%; P = .01). Multiple-birth infants were also at greater risk for developing pneumonia (24% vs 6%; P = .05) and requiring hospitalization (32% vs 18%; P = .05) than were singletons. Additional risk factors for developing pneumonia and bronchiolitis were examined in all 68 children. Multiple birth (P = .05), gestational age of less than 30 weeks (P = .02), and crowded homes (defined as more than one person living in 19 m2 of living space [P = .002] or more than one child living in 22 m2 of living space [P = .004]) were additional risk factors for developing pneumonia. CONCLUSION: Multiple-birth preterm infants are at a higher risk of developing pneumonia than are singletons. Additional risk factors for developing pneumonia in preterm infants with bronchopulmonary dysplasia include gestational age of less than 30 weeks and crowded homes. At-risk infants with any of these risk factors should be targeted for prophylactic and therapeutic interventions against respiratory syncytial virus.

Birth Weight↗