An elongated segment of DNA observed between two human α globin genes

Summary Detailed restriction enzyme analysis of the DNA from a Chinese female showed that one of her chromosomes had a >17.5 kb deletion of DNA, including the , 2, and 1 globin genes, which is present in many Southeast Asians with an -thalassemia-1 chromosome. Her normal chromosome had the expected cluster of -like globin genes (5----2-1-3), but the segment of DNA between the two globin genes was elongated by some 0.5–0.7 kb. Analyses of various restriction sites suggested that this normal variant of the human globin gene complex is due to a crossover between a normal chromosome with () and a chromosome with an -thalassemia-2 (–^3.7) and an -21-hybrid gene.     

Conformational dynamics in mixed alpha/beta-oligonucleotides containing polarity reversals: a molecular dynamics study using time-averaged restraints

Nucleic acid duplexes featuring a single alpha-anomeric thymidine inserted into each DNA strand via 3-3 and 5-5 phosphodiester linkages exhibit local conformational dynamics that are not adequately depicted by conventional restrained molecular dynamics (rMD) methods. We have used molecular dynamics with time-averaged NMR restraints (MDtar) to explore its applicability to describing the conformational dynamics of two -containing duplexes – d(GCGAAT-3-3-T-5-5-CGC)₂ and d(ATGG-3-3-T-5-5-GCTC)r(gagcaccau). In contrast to rMD, enforcing NOE-based distance restraints over a period of time in MDtar rather than instantaneously results in better agreement with the experimental NOE and J-data. This conclusion is based on the dramatic decreases in average distance and coupling constant violations (d _av, J _rms, and J _av) and improvements in sixth-root R-factors (R ^x). In both duplexes, the deoxyribose ring puckering behavior predicted independently by pseudorotation analysis is portrayed remarkably well using this approach compared to rMD. This indicates that the local dynamic behavior is encoded within the NOE data, although this is not obvious from the local R ^x values. In both systems, the backbone torsion angles comprising the 3-3 linkage as well as the (high S-) sugars of the -nucleotide and preceding residue (–1) are relatively static, while the conformations of the 5-5 linkage and the sugar in the neighboring -nucleotide (+1) show enhanced flexibility. To reduce the large ensembles generated by MDtar to more manageable clusters we utilized the PDQPRO program. The resulting PDQPRO clusters (in both cases, 13 structures and associated probabilities extracted from a pool of 300 structures) adequately represent the structural and dynamic characteristics predicted by the experimental data.     

Ribonuclease activity dependent cytotoxicity of Asp fl,a major allergen of A. fumigatus

Vasoactive peptides such as angiotensin II (AII), atrial natriuretic peptide (ANP) and vasopressin play an important role in the regulation of blood pressure. We have recently shown an augmentation of Gi levels in heart and aorta from genetic and experimentally-induced hypertensive rats, which may be attributed to the increased levels of vasoactive peptides. We have therefore investigated the effect of AII and ANP on the expression of G-proteins (Gi and Gs) in cultured vascular smooth muscle cells (VSMC) and their relationship with adenylyl cyclase activity. Exposure of VSMC with AII resulted in the augmentation of the levels of Gi-2 and Gi-3 proteins and Gi-2 and Gi-3 mRNA and not of Gs as determined by immunoblotting and Northern blotting techniques respectively. However, the stimulatory effects of N-ethylcarboxamide adenosine (NECA) and isoproterenol on adenylyl cyclase was diminished by AII treatment, whereas the inhibitory effects of AII and C-ANP4-23 were completely attenuated. On the other hand, pretreatment of the cells with C-ANP4-23 resulted in the reduction of the levels of Gi-2 and Gi-3 and not of Gs. The inhibitory responses of adenylyl cyclase to C-ANP4-23 and AII were also attenuated and the stimulatory effects of GTPgS and other agonists were significantly augmented. These data indicate that AII and ANP modulate the expression of Gia protein in a different manner. It may be suggested that the enhanced levels of Gi protein observed in hypertension may be attributed to the augmented levels of AII and not to ANP.     

Pitiriasis versicolor por malassezia ovalis

In a randomly collected series of 175 cases of pityriasis versicolor in residents of the central-northern part of Venezuela, 13% of the patients were infected with ovoid unicellular units of the causal fungus (Malassezia ovalis) and 87% with globous unicellular units in the scales (M. furfur). Only globous unicellular units (Pityrosporum orbiculare) were seen outside the active lesions and in these, after the successful treatment with ketoconazole. The median age of 40 ovalis patients was 37.5 years; the same in 40 furfur patients was 24.5 years. The median age at the moment of discovering the first lesions among ovalis patients was 31; among furfur patients this was 20 years. There were 16 male patients among the ovalis and 24 among the furfur groups of 40. The topographic distribution of the lesions varied according to the type of the invader. M. ovalis prevailed on the trunk below the waist-line and on the limbs, mainly on buttocks and upper legs. M. furfur prevailed on the chest, neck, face and upper limbs.The hypothesis is offered that the ovoid agent of pityriasis versicolor preferentially occupies the less sweating and sebum-producing parts of the body in older (and drier) persons than the globous type does.     

The sup8 tRNALeu gene of Schizosaccharomyces pombe has an unusual intervening sequence and reduced pairing in the anticodon stem

Summary We have cloned and sequenced the wild-type and suppressor alleles of the S. pombe sup8 tRNA gene. The wild-type allele has a leucine UAA anticodon and the suppressor (sup8-e) carries the opal suppressor anticodon UCA. The gene has a 16 base pair intervening sequence that, in the RNA, is predicted to form a secondary structure which involves base pairing to the 5, rather than the usual 3 side of the 5 splice site. When incubated in Saccharomyces cerevisiae cell-free extracts both alleles are efficiently transcribed, the 5 leader and 3 trailer sequences are removed and CCA is added to the 3 processed end; however, the intervening sequence is not excised. This finding implies that the structural requirements of the splicing endonucleases in the two yeasts have diverged. No other tRNA genes with related sequences were detected in S. pombe DNA by hybridization, suggesting that other UUA isoacceptors may be structurally dissimilar to sup8 or that the UUA codon may be decoded by a UUG leucine isoacceptor.     

Cytological observations on some species and hybrids ofVaccinium

Summary Meiosis was studied inVaccinium myrtillus (n=12),V. vitis-idaea (n=12),V. myrtillus ×vitisidaea (n=12), colchicine-induced tetraploidV. myrtillus (n=24),V. uliginosum (n=24), the cultivated blueberry varieties Rancocas and Pemberton (n=24), and the hybridsV. uliginosum × Rancocas andV. uliginosum × Pemberton (n=24). Pairing was regular in the diploid species. Some multivalent formations occurred inV. uliginosum and in the cultivated blueberry varieties and to a considerable extent in the autotetraploidV. myrtillus. The spontaneous diploid hybridV. myrtillus ×vitis-idaea displayed numerous meiotic irregularities, whereas the artificial hybrids betweenV. uliginosum and cultivated blueberry showed relatively regular meiosis. Pairing relationships of the various genomes are discussed. Breeding programs for the use of North-European species with American cultivated blueberry varieties are discussed in the light of the cytological observations.

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Zusammenfassung Die diploiden (n=12) ArtenVaccinium myrtillus, V. vitis-idaea, V. myrtillus×vitis-idaea und die tetraploiden (n=24)V. myrtillus (Colchicin-induziert) undV. uliginosum sowie die kultivierten amerikanischen Blaubeersorten Rancocas und Pemberton (n=24) und die HybridenV. uliginosum×Rancocas undV. uliginosum×Pemberton wurden cytologisch untersucht. In den diploiden Arten war die meiotische Chromosomenpaarung regelmäßig. Multivalente traten in geringerem Umfange beiV. uliginosum und kultivierten Blaubeersorten, viel stärker aber in der autotetraploidenV. myrtillus auf. Die spontane diploide HybrideV. myrtillus×vitis-idaea zeigte zahlreiche meiotische Störungen, die künstlichen Hybriden zwischenV. uliginosum und der kultivierten amerikanischen Blaubeere hatten demgegenüber eine relativ regelmäßige Meiose. Die Paarungsverhältnisse der verschiedenen Genome werden diskutiert. Auf Grund der cytologischen Beobachtungen wird ein Züchtungsprogramm für nordeuropäischeVaccinium-Arten zusammen mit amerikanischen Kultursorten erörtert.

     

Confirmation of assignment of the human α1-crystallin gene (CRYA1) to chromosome 21 with regional localization to q22.3

Summary The crystallins are highly conserved structural proteins universally found in the eye lens of all vertebrate species. In mammals, three immunologically distinct classes are present, -, -, and -crystallins, and each class represents a multigene family. The -crystallin gene family consists of 1-crystallin (CRYA1) and 2-crystallin (CRYA2) genes (previously designated A-and B-crystallin, respectively), which show extensive sequence homology. We constructed a synthetic oligonucleotide probe of 25 bases corresponding to a specific region of the human 1-crystallin gene sequence. This 25-mer probe bears little sequence homology to human 2-crystallin gene and does not cross-hybridize to 2-crystallin sequences in Southern blot analysis. Using this unique synthetic probe, we have demonstrated the identity of the 1-crystallin gene in human genomic DNA. In addition, we have also confirmed its chromosomal location on human chromosome 21. Finally, we have regionally localized the gene to q22.3 by using both Southern blot analysis of a panel of cell hybrids containing different parts of human chromosome 21, and in situ hybridization to metaphase chromosomes. The use of synthetic oligonucleotide probes specific for individual genes should be useful in identifying and mapping members of multigene families.     

Growth vigour of some Egyptian cotton variety seedlings in relation to selected rhizospheric microflora antagonistic toRhizoctonia solani Kuhn

Summary Growth responses of Ashmouni and Karnak cotton variety seedlings toRhizoctonia solani, the damping-off fungus, or toBacillus subtilis (two different strains),Aspergillus terreus andAspergillus flavus isolated from the rhizosphere of cotton, and all antagonistic to the pathogen, were expressed in terms of growth-vigour criteria.The presence ofR. solani in the soil inhibited the growth vigour of both cotton variety seedlings. However, Karnak seedlings were more sensitive to the pathogen than Ashmouni seedlings. One of the strains ofB. subtilis andA. terreus generally increased the vigour of both cotton variety seedlings.A. flavus lowered most of the growth criteria of Karnak or Ashmouni cotton seedlings.

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Zusammenfassung Rhizoctonia solani, der Parasiet von Baumwolle-Keimlingen, wurde isoliert.Sowohl Bacillus subtilis als auchAspergillus terreus und Aspergillus flavus wurden von der Rhizosphere der Baumwolle-Pflanzen isoliert. Diese Organismen wurden als antagonistisch gegenRhizoctonia solani erkannt. Die Wirkung dieser Organismen auf das Wachstum von Keimlingen der Baumwolle sorten Ashmouni und Karnak wurde untersucht. R. solani hemmt das Wachstum der Keimlinge beider Baumwolle-Sorten. Es wurde festgestellt, dass Karnak empfindlicher ist als Ashmouni. Einer der Stämme vonB. subtilis undA. terreus erwiesen sich als Wachstum förderend bei beiden Baumwolle-Sorten.A. flavus dagegen vermindert das Wachstum von Karnak und Ashmouni.

     

Topography of the subunits of Micrococcus lysodeikticus F1-ATPase

Summary The combined use of proteolytic digestion and lactoperoxidase catalyzed labelling with [¹²⁵I] applied to membrane-bound or soluble pure F₁-ATPase from Micrococcus lysodeikticus has allowed us to establish the topography of its , , and subunits within the protein molecule and with respect to the plane of the membrane.The subunit is most externally located to the membrane bilayer looking towards the cytoplasmic face, a position consistent with its proposed catalytic role. The and subunits lie in an intermediate layer between the subunits and the membrane, in which the subunit occupies a central position within the F₁-ATPase molecule in contact with the subunit. The subunit appears to be tightly bound to the F₀ component of the ATPase complex, probably buried in the membrane bilayer. A molecular arrangement of M. lysodeikticus ATPase is proposed that, taking into account the subunit stoichiometry _{3 3 2 2} (MW 420 000), accommodates the role assigned to each subunit and most, if not all, the known properties of this bacterial energy-transducing protein.     

Purification and characterization of the 5′ → 3′ exonuclease domain-deleted Thermus filiformis DNA polymerase expressed in Escherichia coli

The gene encoding 5 3 exonuclease domain-deleted Tfi DNA polymerase, named 5 3 Exo^– Tfi fragment, from Thermus filiformis was expressed in Escherichia coli under the control of the tac promoter on a high-copy plasmid, pJR. The expressed enzyme was purified 27-fold with a 19% yield and a specific activity of 2621 U mg^–1 protein. The 5 3 exonuclease domain of Tfi DNA polymerase was removed without significant effect on enzyme activity and stability. PCR conditions for the 5 3 Exo^– Tfi fragment were more tolerant to the buffer composition as compared to the full-length enzyme.     

Genotypic stability analysis of yield and related agronomic characters in wheat-Agropyron derivatives under varying watering regimes

Summary Genotypic stability of grain yield and yield related traits was estimated for the 8 wheat-Agropyron derivatives, 441, 442, Vostock, PPG 56, PPG 56SS, 449, 450 and 441SS, together with two spring wheat controls, Drabant and Sv 01382, grown under varying watering regimes for two years. Differences among environments, genotypes and their interactions were highly significant for all traits. The genotypes X environments linear response were significant for number of spikelets per main spike, grain number per main spike and single grain weight. No significant linear responses were detected for grain yield or the rest of yield related traits. Deviations from linear responses were highly significant for all traits under study. Grain yield stability was not associated with the stability of yield components. PPG 56SS was identified as the stable variety with the highest mean grain yield per plant under the severe stress environment. Mean grain yields in the severe stress environment were correlated with the genotypic stability parameter . The severe stress environment was recognized as the optimum test environment. The study revealed a shift in the association between grain yield per plant and its components under the varying environments.     

AMP synthesis in aqueous solution of adenosine and phosphorus pentoxide

Possible formation of a P₄O₁₀ molecule in magma, the stability of the molecule in hydrous volcanic gas at high temperatures and a possible prebiotic phosphate cycle were discussed in relation to chemical evolution. To demonstrate the utility of phosphorus pentoxide as a phosphorylating agent, aqueous solutions of adenosine (0.02M) and phosphorus pentoxide (0.2M) were incubated at 37°C for 5 months. The pH of the solutions was adjusted every day or every few days to each fixed value (9.0, 10.5, 11.5, 12.5) with 10 N NaOH. The HPLC analysis showed the formation of 2-AMP, 3-AMP, 5-AMP, cyclic (2–3)-AMP and cyclic (3–5)-AMP. The main components of the products were 2- and 3-AMP, though cyclic (2–3)-AMP was the main component in the early period of the incubation at pH 9.0. The yields (conversion rate of adenosine to AMPs) were increased almost linearly with the incubation time for 5 months in the case of pH 9.0. The final yields were about 3% (pH 9.0), 6% (pH 9.0, 1 M NaCl), 5% (pH 9.0, 0.01 M CaCl₂, 0.01 M MgCl₂), 7% (pH 9.0, 0.5 M NaCl, 0.01 M CaCl₂, 0.01 M MgCl₂), 9% (pH 9.0, 1 M NaCl, 0.01 M CaCl₂, 0.01 M MgCl₂), 32% (pH 10.5), 43% (pH 11.5), 35% (pH 12.5).     

Inheritance of resistance to potyviruses in Phaseolus vulgaris L. II. Linkage relations and utility of a dominant gene for lethal systemic necrosis to soybean mosaic virus

Summary A single dominant factor, Hss, that conditions a rapid lethal necrotic response to soybean mosaic virus (SMV) has been identified in Phaseolus vulgaris L. cv. Black Turtle Soup, line BT-1. Inoculated plants carrying this factor developed pinpoint necrotic lesions on inoculated tissue followed by systemic vascular necrosis and plant death within about 7 days, regardless of ambient temperature. BT-1 also carries dominant resistance to potyviruses attributed to the tightly linked or identical factors, I, Bcm, Cam, and Hsw, so linkage with Hss was evaluated. No recombinants were identified among 381 F₃ families segregating for potyvirus susceptibility, thus if Hss is a distinct factor, it is tightly linked to I, Bcm, Cam, and Hsw. BT-1 was also crossed reciprocally with the line Great Northern 1140 (GN 1140) in which the dominant gene, Smv, for systemic resistance to SMV was first identified. Smv and Hss segregated independently and are co-dominant. The (GN 1140 x BT-1) F₁ populations showed a seasonal shift of the codominant phenotype. Evaluation of the (GN 1140 x BT-1) F₂ population under conditions where Smv is partially dominant allowed additional phenotypic classes to be distinguished. Pathotype specificity has not been demonstrated for either Smv or Hss. Genotypes that are homozygous for both dominant alleles are systemically resistant to the virus and in addition show undetectable local viral replication or and no seed transmission. This work demonstrates that a gene which conditions a systemic lethal response to a pathogen may be combined with additional gene(s) to create an improved resistant phenotype.     

Identification and confirmation of three neutral alleles conferring wide compatibility in inter-subspecific hybrids of rice (Oryza sativa L.) using near-isogenic lines

Wide-compatibility varieties (WCVs) are a special class of rice germplasm that is able to produce fertile hybrids when crossed to both indica and japonica subspecies. Previous studies determined Dular and 02428 as two WCVs and identified a number of QTLs as having large effects on fertility of inter-subspecific hybrids. In this study, we developed five near-isogenic lines (NILs) for three of the QTLs, f5, f6 and S5, by backcrossing and marker-assisted selection, using Dular and 02428 as the donors and Zhenshan 97 as the recipient. Three of the NILs each carried one introgressed allele, and two NILs each carried two introgressed alleles in combinations. The NILs were testcrossed to an indica tester Nanjing 11 and a japonica tester Balilla. The results showed that the f5 allele from Dular (f5-Du) is a neutral allele conferring wide compatibility, with a large effect on both pollen and spikelet fertility, and the f6 allele from Dular (f6-Du) is a neutral allele for spikelet fertility with smaller effect. The S5 allele from 02428 (S5-08) was confirmed to be a neutral allele for spikelet fertility. It is likely that f6 and S5 are the same locus as deduced by their genomic locations and effects. The results also showed that even in combination, two neutral alleles of different loci were not able to produce normal fertility hybrids in typical indica–japonica crosses. The implications of the findings in rice breeding programs are discussed.     

Keratella mexicana sp. nov., a new planktonic rotifer from Aguascalientes,Mexico

A new species of Keratella is described from a small reservoir in Aguascalientes, Mexico. The species appears related to K. slacki Brziss, 1963 and K. lenzi Hauer, 1953.     

Hexahydroindanone derivatives of steroids formed by Rhodococcus equi

Summary A mutant strain of Rhodococcus equi accumulates three metabolites from the androst-4-ene-3,17-dione or from its degradation intermediate, 3a-H-4(3'-propionic acid)-7a-methylhexahydro-1,5-indanedione (MEPHIP). These three metabolites are: 3a-H-4a(3'-propionic acid)-5-hydroxy-7a-methylhexahydro-1-indanone--lactone (HIL); 3a-H-4(3'-trans acrylic acid)-5-hydroxy-7a-methylhexahydro-1-indanone (^2'-5-hydroxy-MEPHIP); and 3a-H-4(3'-hydroxy-3'-propionic acid)-5-hydroxy-7a-methylhexahydro-1-indanone (3'-hydroxy-HIL). The behaviour of this mutant allows us to propose a pathway for degradation of the intermediates, methylperhydroindanone propionates. However, during this degradation, the side-chain propionate was eliminated by a-oxidation mechanism. Offprint requests to: A. Miclo     

Bacteriophage lambda replication proteins: formation of a mixed oligomer and binding to the origin of lambda DNA

Summary The purified bacteriophage replication proteins O and P sediment separately in metrizamide gradients of low ionic strength as dimers. Together they interact with each other forming an oligomer, composed of two molecules of O and one molecule of P. The O-P oligomer is active in the in vitro replication of ori-containing DNA.Equilibrium sedimentation in preformed metrizamide density gradients under conditions that separate DNA-protein complexes from free proteins was employed in order to study possible interactions among the replication proteins and ori DNA. It was found that the P protein binds specifically to ori-containing plasmid DNA only in the presence of O protein. About 100 molecules of O and 10 molecules of P form a complex with the ori DNA. The DNA-O-P complex was shown to be active in an in vitro replication system.Since the physical interactions between ori and O and between P and the Escherichia coli dnaB replication protein are well documented, the evidence for a O-P interaction presented in this paper provides the missing link in the molecular mechanism that enables to direct the host replication machinery to the replication of its own DNA.     

Formation of α- and β-type keratin in lizard epidermis during the molting cycle

Summary The epidermis of Anolis carolinensis is renewed periodically by molting. Prior to the molt the distinct layers of the epidermis, namely, the Oberhäutchen, , mesos, and layers, are formed in sequence from a morphologically homogenous population of basal cells. The Oberhäutchen, the first cell layer to form, has spinules on the surface which interdigitate with the overlying cells of the clear layer. The cells of the Oberhäutchen develop 80 Å filaments similar to those in the cells of the layer. Beneath the Oberhäutchen is the layer, the cells of which develop membrane-bounded packets containing a homogenous material during the early stages of differentiation. Later 100–500 Å thick fibrils are formed in the membranebounded packets. The fully keratinized cells, however, are packed with filaments 30 Å in diameter separated by an electron dense amorphous matrix, very similar to -type keratin found in the feather rachis. The cells of the layer, which is immediately below the layer, contain 80 Å filaments very similar to the -type keratin found in hair cortex and keratinizing stratified epithelia of mammals. Large quantities of glycogen are found in the cells of each layer during their genesis. Even though a stratum granulosum is not found underneath the layer, the cells of the clear layer develop bodies which have histochemical and ultrastructural characteristics of keratohyalin granules. The old epidermis is then shed in toto at the junction of the clear layer (above) and the Oberhäutchen (below).This study constitutes publication No. 406 from the Oregon Regional Primate Research Center, supported in part by postdoctoral training fellowship 1-TIAM-5521-02 and NIH Grant No. FR-00163.     

T cells in murine lupus: propagation and regulation of disease

MRL/Mp-lpr/lpr mice develop a spontaneous lupus syndrome, including hypergammaglobulinemia, autoantibodies, glomerulonephritis, and lymphadenopathy. To investigate the role of lymphocyte subsets in the pathogenesis of disease, lupus-prone MRL mice deficient in T cells, T cells, or both were generated. Mice deficient in T cells developed a partially penetrant lupus syndrome, characterized by lymphadenopathy, elevated levels of class-switched immunoglobulins, an increased incidence of antinuclear antibodies, and immune deposits in kidneys which progressed to renal insufficiency over time. In comparison to wild type animals, T cell-deficient animals developed an accelerated and exacerbated disease phenotype, characterized by accelerated hypergammaglobulinemia and enhanced autoantibody production and mortality. Repertoire analysis of these latter animals identified polyclonal expansion (V) of CD4+B220-cells. Mice lacking both and T cells failed to generate class-switched autoantibodies and immune complex renal disease. First, these findings demonstrate that murine lupus in the setting of Fas-deficiency does not absolutely require the presence of T cells, and they also suggest that a significant basis for MRL/lpr disease, including renal disease, involves T cell-independent, T cell dependent, polyreactive B cell autoimmunity, upon which T cell-dependent mechanisms aggravate specific autoimmune responses. Second, these data indicate that T cells partake in the regulation of systemic autoimmunity, presumably via their effects on CD4+B220-T cells that provide B cell help. Finally, these results demonstrate that MRL/lpr B cells, despite their intrinsic abnormalities, cannot per se cause tissue injury without T cell help.Abbreviations snRNPs small nuclear ribonucleoprotein particles