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Universal iterative strategy for the divergent synthesis of dendritic macromolecules from conventional monomers by a combination of living radical polymerization and irreversible TERminator multifunctional INItiator (TERMINI).

A new synthetic concept named TERMINI that stands for irreversible TERminator Multifunctional INItiator is reported. Suitable combinations of TERMINI and living polymerizations provide access to strategies for the design and synthesis of unprecedented complex molecular and macromolecular architectures from a diversity of commercial monomers. TERMINI represents a masked multifunctional initiator designed to quantitatively and irreversibly interrupt a chain organic reaction or a living polymerization. After demasking, the TERMINI repeat unit enables the quantitative reinitiation, in the presence or absence of a catalyst, of the same or a different living polymerization or a chain organic reaction in more than one direction, thus becoming a branching point. The demonstration of this concept was made by using a combination of metal-catalyzed living radical polymerization (LRP) and (1,1-dimethylethyl)[[1-[3,5-bis(S-phenyl 4-N,N'-diethylthiocarbamate)phenyl]ethenyl]oxy]dimethylsilane as TERMINI, to elaborate a novel iterative divergent method for the synthesis of dendritic macromolecules based on methyl methacrylate (MMA).

Journal Article↗

Retrotransposon-like sequences integrated into the genome of pineapple, Ananas comosus.

Retrotransposon-like sequences have been serendipitously detected in the genome of commercial pineapple, Ananas comosus. The sequence from a 2.6 kb cloned fragment of this element had greatest similarity to the del1 Lilium henryi retrotransposon and the gypsy/Ty3 group of retroelements. The order of the genes from 5' to 3' was reverse transcriptase, ribonuclease H and integrase. The integrase domain contained the amino acid sequence motifs which have been associated with recognition of the long terminal repeats and with the cutting/joining reactions required for integration of similar retroelements into the host genome. The retrotransposon existed as a population of variable sequences which were dispersed throughout the genome of pineapple. Southern hybridisation showed that the retrotransposon had integrated repeatedly into the pineapple genome. The reading frame of the element was not interrupted by stop codons, suggesting that it is still potentially capable of transposing. This is the first report of a retrotransposon in pineapple, which we have called deal (for dispersed element of Ananas).

Amino Acid Sequence↗

Primary structure and genomic organization of the histidine-rich protein of the malaria parasite Plasmodium lophurae.

The nucleotide sequence of a complete genomic clone for the histidine-rich protein of Plasmodium lophurae has been determined. The deduced amino acid sequence of the mature protein shows numerous tandemly repeated units preceded by a signal and a pro peptide. The gene is interrupted by an intron with a separate exon coding for the signal peptide. The signal peptide-encoding exon detects multiple cross-hybridizing sequences in the parasite genome.

Amino Acid Sequence↗

Phase II trial of prolonged continuous infusion of 5-fluorouracil and interferon-alpha in patients with advanced pancreatic cancer. Eastern Cooperative Oncology Group Protocol 3292.

Evidence suggests that interferon-alpha (IFN-alpha) augments the antineoplastic activity of 5-fluorouracil (5-FU) in human adenocarcinoma cell lines in vitro and may enhance the efficacy of 5-FU in patients with advanced colorectal carcinoma. In addition, 5-FU may be more effective when given as a prolonged, continuous i.v. infusion (PCI). The Eastern Cooperative Oncology Group performed a Phase II trial of PCI 5-FU plus IFN-alpha in patients with advanced pancreatic carcinoma. Twenty-six patients with advanced, surgically incurable adenocarcinoma of the pancreas received PCI 5-FU (250 mg/m2 daily for 28 days) in combination with IFN-alpha (5 x 10(6) IU/m2 s.c. thrice weekly). Treatment cycles were repeated 14 days or longer after completion of the previous cycle. Treatment was interrupted prior to day 28 if intolerable toxicity developed, and the dose of 5-FU was reduced in subsequent cycles. Partial response occurred in two of 24 evaluable patients (8%; 95% confidence interval, 0-19%). The majority of the study group (88%) had liver metastases. Patients whose serum lactate dehydrogenase (LDH) was more than twofold elevated developed 5-FU-related toxicity significantly sooner than patients with smaller elevations in serum LDH (9 vs. 22 days; p = 0.003). A similar trend was observed for patients with a more than twofold elevation in serum glutamic-oxaloacetic transaminase (SGOT; 9 vs. 15 days; p = 0.07). In conclusion, PCI 5-FU plus IFN-alpha has minimal activity in patients with advanced pancreatic carcinoma, and elevated serum LDH and/or SGOT may be useful for predicting greater toxicity from 5-FU-based therapy in patients with liver metastases.

Adenocarcinoma↗

Comparative analysis of sequences flanking tet(W) resistance genes in multiple species of gut bacteria.

tet(W) is one of the most abundant tetracycline resistance genes found in bacteria from the mammalian gut and was first identified in the rumen anaerobe Butyrivibrio fibrisolvens 1.230, where it is highly mobile and its transfer is associated with the transposable chromosomal element TnB1230. In order to compare the genetic basis for tet(W) carriage in different bacteria, we studied sequences flanking tet(W) in representatives of seven bacterial genera originating in diverse gut environments. The sequences 657 bp upstream and 43 bp downstream of tet(W) were 96 to 100% similar in all strains examined. A common open reading frame (ORF) was identified downstream of tet(W) in five different bacteria, while another conserved ORF that flanked tet(W) in B. fibrisolvens 1.230 was also present upstream of tet(W) in a human colonic Roseburia isolate and in another rumen B. fibrisolvens isolate. In one species, Bifidobacterium longum (strain F8), a novel transposase was located within the conserved 657-bp region upstream of tet(W) and was flanked by imperfect direct repeats. Additional direct repeats 6 bp long were identified on each end of a chromosomal ORF interrupted by the insertion of the putative transposase and the tet(W) gene. This tet(W) gene was transferable at low frequencies between Bifidobacterium strains. A putative minielement carrying a copy of tet(W) was identified in B. fibrisolvens transconjugants that had acquired the tet(W) gene on TnB1230. Several different mechanisms, including mechanisms involving plasmids and conjugative transposons, appear to be involved in the horizontal transfer of tet(W) genes, but small core regions that may function as minielements are conserved.

Amino Acid Motifs↗

Cloning of a genetically unstable cytochrome P-450 gene cluster involved in degradation of the pollutant ethyl tert-butyl ether by Rhodococcus ruber.

Rhodococcus ruber (formerly Gordonia terrae) IFP 2001 is one of a few bacterial strains able to degrade ethyl tert-butyl ether (ETBE), which is a major pollutant from gasoline. This strain was found to undergo a spontaneous 14.3-kbp chromosomal deletion, which results in the loss of the ability to degrade ETBE. Sequence analysis of the region corresponding to the deletion revealed the presence of a gene cluster, ethABCD, encoding a ferredoxin reductase, a cytochrome P-450, a ferredoxin, and a 10-kDa protein of unknown function, respectively. The EthB and EthD proteins could be easily detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and were induced by ETBE in the wild-type strain. Upstream of ethABCD lies ethR, which codes for a putative positive transcriptional regulator of the AraC/XylS family. Transformation of the ETBE-negative mutant by a plasmid carrying the ethRABCD genes restored the ability to degrade ETBE. Complementation was abolished if the plasmid carried ethRABC only. The eth genes are located in a DNA fragment flanked by two identical direct repeats of 5.6 kbp. The ETBE-negative mutants carry a single copy of this 5.6-kbp repeat, suggesting that the 14.3-kbp chromosomal deletion resulted from a recombination between the two identical sequences. The 5.6-kbp repeat is a class II transposon carrying a TnpA transposase, a truncated form of the recombinase TnpR, and a terminal inverted repeat of 38 bp. The truncated TnpR is encoded by an IS3-interrupted tnpR gene.

Cloning, Molecular↗

Microvascular lesions of the true vocal fold.

Microvascular lesions, also called varices or capillary ectasias, in contrast to vocal fold polyps with telangiectatic vessels, are relatively small lesions arising from the microcirculation of the vocal fold. Varices are most commonly seen in female professional vocalists and may be secondary to repetitive trauma, hormonal variations, or repeated inflammation. Microvascular lesions may either be asymptomatic or cause frank dysphonia by interrupting the normal vibratory pattern, mass, or closure of the vocal folds. They may also lead to vocal fold hemorrhage, scarring, or polyp formation. Laryngovideostroboscopy is the key in determining the functional significance of vocal fold varices. Management of patients with a varix includes medical therapy, speech therapy, and occasionally surgical vaporization. Indications for surgery are recurrent hemorrhage, enlargement of the varix, development of a mass in conjunction with the varix or hemorrhage, and unacceptable dysphonia after maximal medical and speech therapy due to a functionally significant varix.

Adult↗

Body density affects stroke patterns in Baikal seals.

Buoyancy is one of the primary external forces acting on air-breathing divers and it can affect their swimming energetics. Because the body composition of marine mammals (i.e. the relative amounts of lower-density lipid and higher-density lean tissue) varies individually and seasonally, their buoyancy also fluctuates widely, and individuals would be expected to adjust their stroke patterns during dives accordingly. To test this prediction, we attached acceleration data loggers to four free-ranging Baikal seals Phoca sibirica in Lake Baikal and monitored flipper stroking activity as well as swimming speed, depth and inclination of the body axis (pitch). In addition to the logger, one seal (Individual 4) was equipped with a lead weight that was jettisoned after a predetermined time period so that we had a set of observations on the same individual with different body densities. These four data sets revealed the general diving patterns of Baikal seals and also provided direct insights into the influence of buoyancy on these patterns. Seals repeatedly performed dives of a mean duration of 7.0 min (max. 15.4 min), interrupted by a mean surface duration of 1.2 min. Dive depths were 66 m on average, but varied substantially, with a maximum depth of 324 m. The seals showed different stroke patterns among individuals; some seals stroked at lower rates during descent than ascent, while the others had higher stroke rates during descent than ascent. When the lead weight was detached from Individual 4, the seal increased its stroke rate in descent by shifting swimming mode from prolonged glides to more stroke-and-glide swimming, and decreased its stroke rate in ascent by shifting from continuous stroking to stroke-and-glide swimming. We conclude that seals adopt different stroke patterns according to their individual buoyancies. We also demonstrate that the terminal speed reached by Individual 4 during prolonged glide in descent depended on its total buoyancy and pitch, with higher speeds reached in the weighted condition and at steeper pitch. A simple physical model allowed us to estimate the body density of the seal from the speed and pitch (1,027-1,046 kg m(-3), roughly corresponding to 32-41% lipid content, for the weighted condition; 1,014-1,022 kg m(-3), 43-47% lipid content, for the unweighted condition).

Acceleration↗

Eucapnic voluntary hyperventilation as a bronchoprovocation technique: development of a standarized dosing schedule in asthmatics.

A variety of dosing schedules have been reported for the hyperventilation method of broncho-provocation testing. To evaluate the effect of challenge technique on the bronchoconstrictive response, we had 16 subjects perform eucapnic voluntary hyperventilation (EVH) with dry, room temperature gas using four different dosing schedules. The hyperventilation challenge dosages included the following: (1) a target minute ventilation (VE) of 20 x FEV1 for 6 min; (2) a target VE of 15 x FEV1 for 12 min; (3) an interrupted challenge with a target VE of 30 x FEV1 for 2 min repeated 3 times; and (4) a target VE of 30 x FEV1 for 6 min. Challenges 2, 3, and 4 gave identical absolute ventilatory challenges (identical factor FEV1 x minutes) but at different VE dosages or time. Challenges 1 and 4 were of identical length, but different target VE. The mean postchallenge fall in FEV1 was 16.6 +/- 10.9%, 11.0 +/- 8.1%, 19.6 +/- 9.9%, and 26.7 +/- 11.3% for challenges 1, 2, 3, and 4, respectively. The response to an identical EVH challenge (FEV1 x 30 for 6 min) was reproducible when performed on separate days. We conclude that the challenge technique used for hyperventilation testing will have a significant impact on the bronchoconstrictive response and must be taken into account when interpreting study results. Tests may be quantitatively comparable over a narrow range of challenge time and VE. We recommend that a 6-min uninterrupted EVH challenge using dry, room temperature gas at a target VE of 30 x FEV1 be adopted as the "standard" challenge.

Adult↗

[Characteristics of the neuronal activity of structures of the hippocampal formation (subiculum)].

Spontaneous and evoked neuronal activity of subiculum was investigated in chronic unanaesthetized rabbits. Rhythmic components of two different types were observed in the spontaneous activity of 58% of units: 4.5-6.0 cps bursts (13%) and 2.0-3.8 cps (45%). Besides their basic frequency, these types of rhythmic activity were differentiated by several additional criteria. 63% of the cells responded to applied sensory stimuli, among which auditory stimuli were the most effective. Multimodal units were slightly more numerous (59%) than unimodal ones. In 50% of the recorded units responses were of tonic type, in 31%--of phasic type, and in 19%--specific on-off responses were observed. In majority of units with repeated stimulations relatively rapid process of habituation developed after a short incremental phase. Interruption of septal afferents resulted in disappearence of 4.5-6.0 cps (theta) bursts, in reduction in the number of tonic responses and in absence of habituation. Similarity of neuronal characteristics in the field CA1 and subiculum is discussed.

Animals↗

Effect of oral charcoal and urine pH on sotalol pharmacokinetics.

The effects of orally given activated charcoal, sodium bicarbonate and ammonium chloride on the pharmacokinetics of sotalol were studied in 7 volunteers in a randomized, cross-over study. Serum and urine concentrations of sotalol were determined by HPLC up to 72 h. Activated charcoal, 50 g, given within 5 min of sotalol hydrochloride (160 mg), reduced its absorption by 99%. When given in repeated doses from 6 h on, 50 g followed by 12.5 g at 6 h intervals, charcoal shortened the half-life after sotalol in serum from 9.4 +/- 0.4 h to 7.6 +/- 0.3 h and in urine from 9.4 +/- 0.4 h to 7.3 +/- 0.3 h (p less than 0.01). The excretion of unchanged sotalol into urine was 65% of the dose during the control phase but neither acidification nor alkalinization of urine for 3 days did increase its urinary excretion. Thus, activated charcoal seems to be effective in prevention of the absorption of sotalol when ingested without delay. Given in repeated oral doses charcoal moderately increases the rate of sotalol elimination, probably by interrupting its enterohepatic or enteroenteric circulation.

Administration, Oral↗

[Catheter ablation of accessory atrioventricular conduction pathways in all locations--consecutive results of a new curative therapy of paroxysmal supraventricular tachycardia].

Accessory atrioventricular pathways are a frequent cause of paroxysmal supraventricular tachycardias. This study analyses our results with a recently developed therapeutic approach-radiofrequency (RF) catheter ablation. This was applied in 150 consecutive patients (97 men, 53 women, mean age 42 +/- 15 years) with a total of 159 accessory pathways in all locations. All but 4 patients were symptomatic, with a spectrum ranging from palpitations (146 patients), syncope (39 patients) to aborted sudden death (2 patients). Prior to ablation, 115 patients had received long-term treatment with up to 4 antiarrhythmic drugs unsuccessfully. The mean number of applied current pulses was 12 +/- 14, and the mean cumulative procedure duration was 256 +/- 243 minutes, with a cumulative fluoroscopy time of 49 +/- 72 minutes. Patients with left-sided pathways were approached via the retrograde aortic approach in 88/90 cases. One-hundred fifteen patients were treated in a single session, repeat sessions were required in the remaining 35 patients. The predominant sites of interruption of right-sided and left-sided accessory pathways were their atrial and ventricular insertion, respectively. Long-term cure was achieved in 141 patients (94%), non-life-threatening complications were observed in 3 patients (2%). These results compare well with published studies on large patient collectives and demonstrate that RF catheter ablation, which is highly cost effective, is the therapy of choice to cure symptomatic patients with accessory atrioventricular pathways in all locations.

Adult↗

Custom devices for sleep apnea treatment.

Obstructive sleep apnea (OSA) is a potentially life-threatening sleep disorder that is estimated to affect 20 million Americans. OSA is characterized by repeated collapse of the upper airway during sleep, which produces breathing pauses and interruption of the sleep cycle. Symptoms include heavy snoring, many nighttime arousals, sleep deprivation, chronic fatigue and a variety of cardiovascular diseases. While many treatments are advocated for OSA, oral devices have been shown to be effective. Dental technicians and dentists may be involved in the construction of oral appliances for patients with OSA. In this article, a synopsis of various treatment modalities is presented, and the construction of two appliances is described and illustrated.

Dental Casting Technique↗

The site-specific ribosomal DNA insertion element R1Bm belongs to a class of non-long-terminal-repeat retrotransposons.

Two types of insertion elements, R1 and R2 (previously called type I and type II), are known to interrupt the 28S ribosomal genes of several insect species. In the silkmoth, Bombyx mori, each element occupies approximately 10% of the estimated 240 ribosomal DNA units, while at most only a few copies are located outside the ribosomal DNA units. We present here the complete nucleotide sequence of an R1 insertion from B. mori (R1Bm). This 5.1-kilobase element contains two overlapping open reading frames (ORFs) which together occupy 88% of its length. ORF1 is 461 amino acids in length and exhibits characteristics of retroviral gag genes. ORF2 is 1,051 amino acids in length and contains homology to reverse transcriptase-like enzymes. The analysis of 3' and 5' ends of independent isolates from the ribosomal locus supports the suggestion that R1 is still functioning as a transposable element. The precise location of the element within the genome implies that its transposition must occur with remarkable insertion sequence specificity. Comparison of the deduced amino acid sequences from six retrotransposons, R1 and R2 of B. mori, I factor and F element of Drosophila melanogaster, L1 of Mus domesticus, and Ingi of Trypanosoma brucei, reveals a relatively high level of sequence homology in the reverse transcriptase region. Like R1, these elements lack long terminal repeats. We have therefore named this class of related elements the non-long-terminal-repeat (non-LTR) retrotransposons.

Amino Acid Sequence↗

Multiple copies of a retroposon interrupt spliced leader RNA genes in the African trypanosome, Trypanosoma gambiense.

The 140-nucleotide spliced leader (SL) RNA, involved in mRNA maturation in the African trypanosomes and in other kinetoplastida, is encoded by a tandem array of spliced leader genes. We show that the 1.4-kb SL gene repeat unit in Trypanosoma gambiense is organized in tandem arrays confined to two large (minimum size 350-450 kb) restriction fragments. SL genes in both arrays are interrupted by a total of eight conserved insertion elements. Cleavage of genomic DNA at restriction sites present within the insertion element but not in the SL gene repeat, releases variable numbers of SL genes from the tandem array. Since the insertion element contains a terminal poly(A) track of 36 bases and because a 49-bp duplication of target DNA has occurred at the integration site, we conclude that it is a retroposon. This retropson is uniquely associated with the SL gene clusters. These retroposons presumably originated from a single insertion event after which their copy number increased, possibly through unequal sister chromatid exchange.

Animals↗

Interrupter resistance: what's feasible?

Interrupter resistance (Rint) is one of the easiest ways to assess respiratory resistance during tidal breathing with minimal subject cooperation. This article enclosed current knowledge on technical and practical aspects such as how to measure Rint, and how to calculate Rint. Issues on repeatability of the technique and bronchial responsiveness are discussed. Recommendations on Rint technique are provided on behalf of the Interrupter Technique Subcommittee of the ATS/ERS Working Group on Infant and Young Children Pulmonary Function Testing.

Airway Resistance↗

Applicability of interrupter resistance measurements using the MicroRint in daily practice.

This study was performed to evaluate the applicability of a simple device (MicroRint) for measuring airway resistance, to derive normal values and to compare values with maximal expiratory flow volume (MEFV) parameters in asthmatic and healthy children. Repetitive R(int) measurements were performed in 125 healthy children and 107 asthmatic children (age range 0.8-16.8 years). In 42 asthmatic patients R(int) and MEFV values were compared and in 29 asthmatic children bronchodilation testing was performed. Successful R(int) measurements were possible in 91% of the children. The mean coefficient of variation of repeated measurements was 7.1 (+/-6.1)%. R(int) values of healthy children showed a significant curvilinear correlation with age (r=-0.80, P < 0.001) and height (r=-0.81, P < 0.001). In asthmatic and healthy children R(int) values were comparable. A significant inverse correlation was found between R(int) and MEFV values (for FEV1 and R(int) r=-0.80, P < 0.001). After bronchodilation there was a significant increase in FEV1 and decrease in R(int), but changes between the two parameters did not correlate. In conclusion, the interrupter technique is feasible and repeatable in children and has a significant correlation with other parameters of airway caliber. Baseline values do not discriminate healthy from asthmatic children.

Airway Resistance↗

The comparative genomics of polyglutamine repeats: extreme differences in the codon organization of repeat-encoding regions between mammals and Drosophila.

Polyglutamine repeats within proteins are common in eukaryotes and are associated with neurological diseases in humans. Many are encoded by tandem repeats of the codon CAG that are likely to mutate primarily by replication slippage. However, a recent study in the yeast Saccharomyces cerevisiae has indicated that many others are encoded by mixtures of CAG and CAA which are less likely to undergo slippage. Here we attempt to estimate the proportions of polyglutamine repeats encoded by slippage-prone structures in species currently the subject of genome sequencing projects. We find a general excess over random expectation of polyglutamine repeats encoded by tandem repeats of codons. We nevertheless find many repeats encoded by nontandem codon structures. Mammals and Drosophila display extreme opposite patterns. Drosophila contains many proteins with polyglutamine tracts but these are generally encoded by interrupted structures. These structures may have been selected to be resistant to slippage. In contrast, mammals (humans and mice) have a high proportion of proteins in which repeats are encoded by tandem codon structures. In humans, these include most of the triplet expansion disease genes.

Animals↗