Evolution in the laboratory: the genome of Halobacterium salinarum strain R1 compared to that of strain NRC-1.

We report the sequence of the Halobacterium salinarum strain R1 chromosome and its four megaplasmids. Our set of protein-coding genes is supported by extensive proteomic and sequence homology data. The structures of the plasmids, which show three large-scale duplications (adding up to 100 kb), were unequivocally confirmed by cosmid analysis. The chromosome of strain R1 is completely colinear and virtually identical to that of strain NRC-1. Correlation of the plasmid sequences revealed 210 kb of sequence that occurs only in strain R1. The remaining 350 kb shows virtual sequence identity in the two strains. Nevertheless, the number and overall structure of the plasmids are largely incompatible. Also, 20% of the protein sequences differ despite the near identity at the DNA sequence level. Finally, we report genome-wide mobility data for insertion sequences from which we conclude that strains R1 and NRC-1 originate from the same natural isolate. This exemplifies evolution in the laboratory.

A family of halobacterial transducer proteins.

A DNA probe to the signaling domain of a halobacterial transducer for phototaxis (HtrI) was used to clone and sequence four members of a new family of transducer proteins (Htps) in Halobacterium salinarium potentially involved in chemo- or phototactic signal transduction. The signaling domains in these proteins have 31-43% identity when compared with each other or with their bacterial analogs, the methyl-accepting chemotaxis proteins. An additional region of homology found in three of the Htps has 31-43% identity with HtrI. The Htps contain from 0 to 3 transmembrane helices and Western blotting showed that HtpIII is soluble. The arrangement of the domains in these Htps suggests a modular architecture in their construction.

Aprendizaje basado en problemas: logros de aprendizaje en las sub-categorías de información y comprensión del dominio cognitivo de Bloom / Learning based in problems: achievement in learning in sub-categories of information and comprehension of cognitive domain of Bloom

The aim of this work was to assess and compare the achievements of medical students, subjected to problem based learning methodology. The information and comprehension categories of Bloom were tested in 17 medical students in four different occasions during the physiopathology course, using a multiple choice knowledge test. There was a significant improvement in the number of correct answers towards the end of the course. It is concluded that these medical students obtained adequate learning achievement in the information subcategory of Bloom using problem based learning methodology, during the physiopathology course

The primary structure of halocyanin, an archaeal blue copper protein, predicts a lipid anchor for membrane fixation.

Halocyanin, a small blue copper protein, was isolated from the haloalkaliphilic archaeon Natronobacterium pharaonis. The NH2 terminus was not accessible to Edman degradation. About 70% of the amino acid sequence was determined by protein sequence analysis. The sequence information of two peptides was used for cloning and sequencing the halocyanin gene (hcy). The open reading frame codes for 489 base pairs, which account for a protein with 163 amino acids and a molecular mass of 17,223 Da. The discrepancy between this value and the molecular mass of 15,456 +/- 1.5 Da for the copper-free protein determined by electrospray mass spectrometry can be explained by a post-translational processing of the gene product. The NH2-terminal sequence of the open reading frame contains a motif that is characteristic for prokaryotic lipoproteins. Assuming a similar processing for halocyanin, Cys at position 25 of the primary transcript would be modified by a diphytanyl (glycerol)diether. Subsequently, the precursor is cleaved by a signal peptidase II-like protease and then acetylated at its NH2-terminal alpha-amino group. These modifications would yield a protein with a calculated molecular mass of 15,456 Da. A comparison of the primary structure of halocyanin with a number of other blue copper proteins places it into the plastocyanin-related group.

The primary structure of the hemoglobins of a southern hemisphere lamprey (Mordacia mordax, Cyclostomata).

Mordacia mordax is a southern hemisphere lamprey belonging to Mordaciidae, a primitive family of Cyclostomata. Adult erythrocytes contain three monomeric hemoglobins which can be easily separated by cellulose acetate electrophoresis and isolated by ion-exchange chromatography. The N-terminal regions, and the tryptic peptides from each chain were submitted to automated Edman degradation; the alignment of the fragments was obtained by homology with the other Petromyzonoidea hemoglobins hitherto sequenced. Our results confirm the phylogenic distance between lampreys and hag-fish hemoglobins. As was observed for Petromyzon marinus species, two hemoglobins of Mordacia mordax are very close, as they differ only at 7 positions.

The primary structures of gundi (Ctenodactylus gundi, Rodentia) hemoglobin and myoglobin.

The primary structures of the alpha- and beta-chains of hemoglobin and myoglobin from the gundi (Ctenodactylus gundi, Rodentia) are presented. The sequences were determined after enzymatic and chemical cleavages of the polypeptide chains and by sequencing of the peptides mainly by automated sequence analysis. The sequences of gundi hemoglobin chains and of myoglobin are compared with those of other rodents.

Prevalencia de bacio endémico en escolares del centro y sur de Chile / Prevalence of endemic goiter among school children of center and southern Chile

Se estudió la prevalencia de bocio endémico en una muestra representativa de escolares de enseñanza básica fiscal de dos ciudades, Santiago y Temuco, en el año 1986. Esta resultó ser de 7,6% en la muestra total, sin diferencias de acuerdo al área geográfica de procedencia. Se encontró mayor prevalencia en las mujeres, y cuanto mayor era su edad (p < 0,05), lo que no se detectó en los varones. Estos resultados sugieren una importante disminución en la prevalencia del bocio endémico en nuestro país en los últimos cuatro años, pues, en las mismas comunas de Santiago, era 25 y 28,8%, respectivamente, en 1982. Se subraya la conveniencia de mantener los programas de prevención y vigilancia de bocio endémico en Chile

Carnivora: the primary structure of the alpha-chains of ferret (Mustela putorius furo, Mustelidae) hemoglobins.

Ferret erythrocytes contain two hemoglobins differing only by their alpha-chains. The primary structure of the common beta-chain has been previously described; the complete sequence of the two alpha-chains are reported in this paper. The globin chains were separated by ion-exchange chromatography; the alpha-chains (42 steps), their tryptic peptides as well as the prolyl-peptides were subjected to automatic liquid- and gas-phase Edman degradation. The two alpha-chains are very similar, differing at only one position (Asp15----Gly15). Comparison with human hemoglobin alpha-chain shows 16 and 17 exchanges, for alpha 1 and alpha II chains, respectively; two substitutions involve alpha 1/beta 1 contacts and one the heme contacts. A high degree of homology was noted when the alpha-chains were compared to the corresponding chains of other representatives of the Carnivora order.

Carnivora: primary structure of the hemoglobin from the spotted hyena (Crocuta crocuta, Hyenidae).

The primary structure of the alpha- and beta-chains of hemoglobin from spotted hyena (Crocuta crocuta, Hyenidae) is presented. The structure-function relationship is discussed. The separation of the chains directly from hemoglobin was performed by RP-HPLC. After tryptic digestion of the chains, the peptides were isolated by RP-HPLC. Amino-acid sequences were determined by Edman degradation in liquid- and gas-phase sequencers. The alignment of the tryptic peptides was made by homology with human and other Carnivora hemoglobins. The hemoglobin from spotted hyena (Crocuta crocuta) exhibits in its alpha- and beta-chains 22 and 20 exchanges, respectively, compared to human hemoglobin. In the alpha-chains, two alpha 1 beta 1-contacts are exchanged. In the beta-chains five exchanges involve one alpha 1 beta 1-contact, one alpha 1 beta 2-contact, one heme contact, and two 2,3-DPG-binding sites.

Características actuales del bocio endémico en escolares de dos zonas censoras de Chile / Some present characteristics of endemic goiter in school age children of 2 zones of Chile

Se detectó una prevalencia de 7,6% de bocio endémico en una muestra de escolares de educación básica fiscal y se encontró una prevalencia significativamente mayor en las mujeres, y en éstas mayor a mayor edad y desarrollo puberal. No se encontró correlación entre bocio y nivel socioeconómico, estado nutricional, calidad estatural ni localización geográfica de la residencia del escolar. Estos resultados muestran una importante disminución en la prevalencia de bocio comparado con el trabajo realizado en 1982. La pérdida de algunas características del bocio se pueden explicar por la notoria disminución de su prevalencia

Carnivora: primary structure of the hemoglobins from ratel (Mellivora capensis).

The erythrocytes of adult ratel contain two hemoglobin components, with two alpha- and one beta-chains. In this paper, their complete amino acid sequences are presented. The two alpha-chains differ in one residue at position 34 (Ala----Val) only. The primary structure of the chains was determined by sequencing the N-terminal regions (45 steps) and the tryptic peptides after their isolation from the digests by reversed-phase high-performance liquid chromatography. The alignment of these peptides was deduced from homology with other carnivora globins. The alpha-chains show 21 and the beta-chains 11 exchanges compared with human globin chains. In the alpha-chains, one heme- and two alpha 1/beta 1 contacts are exchanged. In the beta-chains there are three exchanges which involve one alpha 1/beta 1-, one alpha 1/beta 2- and one heme-contact. Between the ratel hemoglobin and those of carnivora a high degree of homology was found.

Homeothermic fish and hemoglobin: primary structure of the hemoglobin from bluefin tuna (Thunnus thynnus, Scromboidei).

Some fish are warm-bodied, e.g. the bluefin tuna (Thunnus thynnus), which has a muscle temperature 12-17 degrees C higher than its environment. This endothermy is achieved by aerobic metabolism and conserved by means of a heat-exchanger system. The hemoglobins of bluefin tuna are adapted to these conditions by their endothermic oxygenation, thus contributing to the preservation of the body energy. This is a new and so far unique property of tuna hemoglobin. The primary structure of the alpha and beta chains of bluefin tuna hemoglobins is presented. The sequence was determined after enzymatic and chemical cleavages of the chains and sequencing of the peptides in gas- and liquid-phase sequencers. The alpha chains consists of 143 residues and are N-terminally acetylated. The beta chains have 146 amino acids and show two ambiguities at positions 140 and 142. The alpha chains differ from the human alpha chains in 65 amino-acid residues, the beta chains in 76. The hemoglobins of bluefin tuna, carp and man are compared and their different physiological properties are discussed in relation to the sequence data. From the primary structure of tuna hemoglobins, it is possible to propose a molecular basis for their peculiar endothermic transition from the T to the R structure.

Primary structure of the minor haemoglobins from the sea lamprey (Petromyzon marinus, Cyclostomata).

Erythrocytes of the adult Sea Lamprey Petromyzon marinus contain several haemoglobin species, but only the main constituent has hitherto been sequenced. The present paper describes the determination of the primary structures of the two minor species, whose electrophoretic mobilities are higher and lower than that of the main component. Tryptic peptides from both chains were purified by high-performance liquid chromatography, then sequenced and aligned by homology with the main haemoglobin. The fast and the major components appeared to be very similar, differing in only four positions (pos. 5: Ser----Thr; pos. 33: Thr----Ser; pos. 86: Val----Ala; pos. 99: Gly----Arg). The slow haemoglobin component, however, differed from the main component with respect to 27 amino-acid residues. The position of the three globins of Petromyzon marinus in the phylogenetic tree of haemoglobins is discussed and a relationship with primitive alpha-chains is postulated.

Primary structure of the hemoglobins from the greater Kudu antelope (Tragelaphus strepsiceros).

The adult greater Kudu antelope has two hemoglobin components, Hb A and Hb B, with one alpha and two beta chains. The complete amino-acid sequences of these three chains are presented. The two beta chains differ only in one residue at position 16 (Gly----Ser) and may be the product of two allelic genes. The primary structure of the chains was determined by sequencing the tryptic peptides after their isolation from the tryptic digest of the chains by high performance liquid chromatography. The alignment of these peptides was deduced from homology with the chains of bovine hemoglobin. Between the Kudu hemoglobins and those of cattle a high degree of homology was found.

Primary structure, biochemical and physiological aspects of hemoglobin from South American lungfish (Lepidosiren paradoxus, Dipnoi).

The South American Lungfish has only one hemoglobin component. The complete amino-acid sequence of this hemoglobin is presented. A large quantity of carbonate dehydratase from the lungfish erythrocytes was also isolated. The carboxymethylated chains, obtained by separation of globin on DEAE-Sephacel, were submitted to tryptic digestion and chemical cleavage. The isolation of tryptic peptides was achieved either by Dowex-50 chromatography or by high performance liquid chromatography. The alignment of peptides was performed by homology with the previously established sequences of the carp and goldfish hemoglobins. The overlapping peptides confirmed this sequence. The alpha chains have 143 residues, the beta chains 147. The relation between the primary structure and the physiological properties of lungfish hemoglobin are discussed.

[The primary structure of hemoglobin from goldfish (Carassius auratus)]. / Die Primärstruktur des Hämoglobins vom Goldfisch (Carassius auratus).

The primary structures of the alpha- and beta-chains from goldfish hemoglobin are given. The globin chains were separated by gel filtration after air-oxidation of globin. After chemical and enzymatical cleavage of the chains, the peptides were isolated by gel filtration and ion exchange chromatography on Dowex. The fish-chains have one residue more than the human chains. The alpha-chain is acetylated at the amino-terminal residue and has no cysteine. Compared with the human chains there are 66 amino-acid differences in the alpha- and 72 in the beta-chains. The implication of these differences for the physiology of the hemoglobin molecule of goldfish is discussed.

Haemoglobins, LX. Primary structure of the major haemoglobin of the sea lamprey Petromyzon marinus (var. Garonne, Loire).

Lampreys belong to the class of Cyclostomata; practically no evolution of these Vertebrates can be noted since Paleozoïc times; lampreys thus appear as a choice material for studying several problems in the field of biochemical evolution. Several monomeric haemoglobins can be characterized in the erythrocytes of the sea lamprey (Petromyzon marinus). The major constituent was isolated by chromatography, and submitted to tryptic digestion; soluble tryptic peptides were separated by gel filtration into 5 fractions; the peptides of each fraction were isolated either by Dowex-50 chromatography or by HPLC; the insoluble core was oxidized and submitted to HPLC fractionation. The primary structure of the whole chain and of the purified tryptic peptides was determined using automatic sequencing; alignment of the peptides was achieved by homology with the previously established covalent structure of the globin of Lampetra fluviatilis. The sequence we established confirms the crystallographic data of Hendrickson and Love. Globin/haem contacts are discussed; a tentative explanation of the absence of tetramerization can be proposed after comparison with the aminoacid residues involved in alpha 1 beta 1 and alpha 1 beta 2 contacts. Petromyzon globin differs at three locations (Thr/Ser3, Leu/Met58, Thr/Ser60) from Lampetra fluviatilis globin. The monomeric chain of another Cyclostomata Myxine glutinosa, differs more considerably (88 residues). Our results corroborate recent paleontologic data which favour the separation of lampreys from hagfishes; Cyclostomata cannot be considered as a monophylic group. Finally, there is a closer relation between lamprey globin and alpha chains than between this monomeric globin and beta chains, and furthermore apomyoglobins of higher vertebrates.

[The sequence of the alpha- and beta-chains of the goldfish hemoglobin (Carassius auratus) (author's transl)]. / Die Sequenz der alpha- and beta-Ketten des Hämoglobins des Goldfisches (Carassius auratus).

The primary structures of the alpha- and beta-chains from goldfish hemoglobin are given. The globin chains were separated by gel filtration after oxidation of globin with air. The peptides for sequencing were obtained by chemical and enzymatical cleavage. The alpha-chain has 142 residues, the beta-chain 147. The alpha-chain is acetylated at the amino-terminal residue. Compared with the human chains, there are 66 amino acid differences in the alpha-chain, 72 residues are different in the beta-chain; the alpha-chain has no cysteine.