Non-coordinate synthesis of RNA polymerase beta beta'' subunits in a temperature-sensitive beta''-subunit mutant of Escherichia coli

Summary On exposure to high temperature of a temperature-sensitive RNA polymerase subunit (rpoC92) mutant of Escherichia coli, selective reduction was observed in the rate of synthesis of a group of proteins including RNA polymerase subunit. The finding that the synthesis of subunit but not subunit was specifically repressed in this mutant grown at non-permissive temperature indicates that the functionally intact RNA polymerase is required for the synthesis of subunits be coordinated. In addition, the assembly of newly synthesized RNA polymerase subunits was inefficient in this mutant at the steps where altered subunit was involved, and the unassembled enzyme subunits were rapidly and preferentially degraded. During recovery to non-restricted growth, the synthesis of both and subunits was transiently enhanced in parallel leading to recovery of the intracellular concentration of functional RNA polymerase.     

Zur Paarbildung der Uferschwalbe (Riparia riparia)

Zusammenfassung Uferschwalben kehren aus den afrikanischen Winterquartieren in Trupps beiderlei Geschlechts zurück. Erste Beringungsergebnisse belegen, daß zunächst mehrjährige, vermutlich untereinander bekannte Vögel eintreffen, die den Brutplatz aus vergangenen Brutperioden her kennen. Die Masse der später ankommenden Vögel dürfte weitgehend aus einjährigen oder ortsfremden Uferschwalben bestehen, die sich größtenteils erst während der Paarbildung persönlich kennenlernen. Der anfängliche Schwarmzusammenhalt der nacheinander eintreffenden Trupps führt zur Bildung von Subkolonien, die für Brutplätze ab einer bestimmten Größenordnung typisch sind. Uferschwalben- gründen nacheinander mehrere Reviere, d. h. sie besetzen Steilwandbereiche, in denen sie ausschließlich mit den Füßen eine Röhre oder Mulde graben, singen und Bogenflüge starten. Bis auf singende oder bekannte werden Artgenossen im Revier geduldet. Uferschwalben- suchen besetzte Reviere auf. Ohne Röhrenbindung verhalten sie sich still und unauffällig, ihre Grabungsaktivitäten sind von untergeordneter Bedeutung. Die Bindung an ein bestimmtes Revier entwickelt sich individuell verschieden und entscheidet über den Abschluß des Röhrenbaues (Herstellung der Nistkammer). Reviere ohne dauerhafte -Bindung werden von den aufgegeben. Aktivitäten, die auf wachsende Revierbindung eines hindeuten, sind: häufige oder/und länger dauernde Aufenthalte des in einem besetzten Revier und sporadisches Mitgraben; aggressives Verhalten gegenüber Artgenossen (i. d. R. fremde ), die im Revier landen wollen; gemeinschaftlicher, leiser Gesang von und im Röhrenbereich. Aktivitäten, die für eine vollzogene Paarbildung sprechen, sind: Fertigstellen der Röhre durch Grabung der Nestkammer; längere gemeinsame Aufenthalte innerhalb und außerhalb der Röhre; Voranfliegen des beim Röhrenanflug; Übernachten von und in der Röhre; Nestbau; ausdauernde Verfolgungsflüge während der Kopulationsphase. Die Paarbildung ist demnach ein individueller Prozeß, bei dem die Aktivitäten der im Revier als Werbung, die der als Revierwahl interpretiert werden.

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On pair-formation in the Sand Martin,Riparia riparia

Summary European Sand Martins arrive at their breeding sites in flocks of usually unmated and . Ringing results of a large population in NW-Germany and own observations indicate that the first flocks about a dozen individuals with an approximately balanced sex ratio appear at traditional breeding places and consist of older, experienced resident birds (presumably acquainted with one another). The birds arriving over the next several weeks are mainly first-year or non-resident individuals. The flocks arrive separately in areas with suitable sandcliffs, synchronize the pair-formation activities and avoid disturbances among paired and unpaired birds. This behaviour causes the formation of subcolonies, which are typical for all densely occupied breeding places. Each settles on a fixed area on the sandcliff (territory) in order to excavate a burrow, to sing the territory-song (fig. 2 b) and to perfor the territory-circle-flight (fig. 2 c, 4 a). Silent birds (normally ) are welcomed or tolerated by the resident . The sexes are monomorphic and therefore courtship displays of the are non-aggressive until establishment of pair-bonds. Only intruding singing or individually known neighbouring are driven away, usually at early stages of territory occupation. Unmated are normally shy and very sensitive to protracted disturbances. visit several occupied territories of the colony (fig. 1–3) in order to choose a burrow. leave territories which do not attract a . They settle new territories on the sandcliff, causing a surplus of burrows compared to breeding pairs in the colony. Activities which indicate the development of pair-bonds are: regular visits of a to a particular occupied territory with sporadic excavations by the ; aggressive activities of the towards other visitors usually , but sometimes at first even against the resident (i. e.: vocal threats, bill-gaping, pecking or pushing with the bill or vigorous face-to-face fights, fig. 3 b, 3 c). and sing the soft mating song at or in the burrow (fig. 1 c). Activities which indicate completed pair-bonds are: completion of the burrow by digging the nestchamber, predominantly done by the ; both birds staying together over long periods, both inside and outside the burrow; invitation-flight by the (fig. 4 b); and spending the night together in the burrow; beginning of nest-building, first only by the , then by both birds and finally only by the , accompanied by the (guarding-flight;, fig. 4c); mate-pursuit flights (sexual chases) during copulation phase, in which the singing pursues the silent , often accompanied by other (cp. fig. 4 d). Pair-formation in the Sand Martin occurs on individual territories and not, according toHickling (1959), within the flock.

     

Net N input and riverine N export from Illinois agricultural watersheds with and without extensive tile drainage

Some of the largest riverine N fluxes in the continental USA have been observed in agricultural regions with extensive artificial subsurface drainage, commonly called tile drainage. The degree to which high riverine N fluxes in these settings are due to high net N inputs (NNI), greater transport efficiency caused by the drainage systems, or other factors is not known. The objective of this study was to evaluate the role of tile drainage by comparing NNI and riverine N fluxes in regions of Illinois with similar climate and crop production practices but with different intensities of tile drainage. Annual values of NNI between 1940 and 1999 were estimated from county level agricultural production statistics and census estimates of human population. During 1945–1961, riverine nitrate flux in the extensively tile drained region averaged 6.6kgNha^–1year^–1 compared to 1.3 to 3.1kgNha^–1 for the non-tile drained region, even though NNI was greater in the non-tile drained region. During 1977–1997, NNI to the tile-drained region had increased to 27kgNha^–1year^–1 and riverine N flux was approximately 100% of this value. In the non-tile-drained region, NNI was approximately 23kgNha^–1year^–1 and riverine N flux was between 25% and 37% of this value (5 to 9kgNha^–1year^–1). Denitrification is not included in NNI and, therefore, any denitrification losses from tile-drained watersheds must be balanced by other N sources, such as depletion of soil organic N or underestimation of biological N fixation. If denitrification and depletion of soil organic N are significant in these basins, marginal reductions in NNI may have little influence on riverine N flux. If tile drained cropland in Illinois is representative of the estimated 11 million ha of tile drained cropland throughout the Mississippi River Basin, this 16% of the drainage area contributed approximately 30% of the increased nitrate N flux in the Lower Mississippi River that occurred between 1955 and the 1990s.     

Molecular switch of F0F1-ATP synthase,G-protein,and other ATP-driven enzymes

Exchange-out of amide tritium from labeled -subunit of ₃₃ complex of F₀F₁-ATP synthase was not accelerated by ATP, suggesting that hemagglutinin-type transition of coiled-coil structure did not occur in -subunit. Local topology of nucleotide binding site and switch II region of G-protein resemble those of F₁- subunit and other proteins which catalyze ATP-triggered reactions. Probably, binding of nucleotide to F₀F₁-ATP synthase induces conformational change of the switch II-like region with transforming subunit structure from open to closed form and this transformation results in loss of hydrogen bonds with the subunit, thus enabling the subunit to move.     

Effect of oxygen on growth and respiration of potato tuber callus

Growth and oxygen uptake of potato callus is faster in an oxygen-enriched atmosphere (70% oxygen, v/v; oxygen-callus) than in air (20% oxygen, v/v; air-callus). Especially the non-mitochondrial, so-called residual respiration is increased in oxygen-callus. The capacities of the mitochondrial respiratory pathways (cytochrome pathway, V_cyt and alternative pathway, V_alt) are also higher in this callus. In both callus types only a small part of the alternative pathway capacity is used in uninhibited respiration. The lower oxygen uptake of air-callus at normal air oxygen pressures is partially due to diffusional impedance. Measurement of the respiratory parameters of air-callus in oxygen-saturated medium leads to higher values than measurement in air-saturated medium, although these values are still lower than those of oxygen-callus.ATP-production was calculated from the oxygen-uptake data and compared with the dry weight production of the callus to give values of 10.0 and 10.8 g dry weight produced.-mol ATP^-1, for air-callus and oxygen-callus respectively. As no harmful side-effects are observed, cultivation of callus under elevated oxygen pressures may be useful, when rapid callus-growth is necessary.Abbreviations AA antimycin A; A; - BHAM benzohydroxamate - DW dry weight - FW fresh weight - 8-OHQ 8-hydroxyquinolin - RC respiratory control - SHAM salicylhydroxamate     

Cajaflavanone and cajanone released from Cajanus cajan (L. Millsp.) roots induce nod genes of Bradyrhizobium sp.

A broad-host-range plasmid (pEA2-21) containing a Bradyrhizobium sp (F-4) nod DABC-lacZ translation fusion was constructed and used to monitor nod gene expression in response to pigeonpea root exudate. Two nod-inducing compounds were isolated and identified. Spectral analysis using ultraviolet absorption, infrared spectra, proton nuclear magnetic resonance, and mass spectrometry showed that the two inducers were 5,4-dihydroxy-6-(3-methyl-2-butenyl)-2, 2-dimethyl pyrano-[5, 6:7, 8]-flavanone (cajaflavanone) and 2,4,5-trihydroxy-5-isopentenyl-6, 7-dimethylchromene iso-flavanone (cajanone). When pEA2-21 was introduced into Rhizobium trifolii and R. meliloti cajanone and cajaflavanone did not induce nod gene indicating that specificity of induction appears to be influenced by the host-strain genome.     

Gonadectomy induces laminin biosynthesis and basement membrane assembly in anterior pituitary glands of adult rats

Summary Laminin biosynthesis and basement membrane assembly in anterior pituitary glands of gonadectomized rats were studied by immuno-electron microscopy and radioimmunoassay. Three weeks after gonadectomy, rats received intravenous injections of sheep anti-laminin IgG conjugated to horseradish peroxidase, and glands were fixed and processed for microscopy 1 h later. Peroxidase reaction product uniformly labeled all perivascular and glandular epithelial basement membranes. In addition, reaction product was also found in abnormally multi-layered basement membranes seen especially beneath gonadotrophs, and unusual basement membrane-like structures projecting between gonadotrophs were also labeled. Pituitary sections from gonadectomized rats labeled with pre-embedding immunoperoxidase and post-embedding immungold techniques also localized intracellular laminin within biosynthetic organelles and light body vesicles of gonadotrophs. Neither abnormal basement membrane structures nor intracellular laminin were detected in pituitaries of nongonadectomized, control rats. Radioimmunoassays of pituitary homogenates showed nearly twice as much soluble laminin ( 15 ng/gland) in gonadectomized rats than in controls ( 8 ng/gland), which paralleled gland growth, but serum laminin concentrations did not differ ( 10 ng/ml in both groups). When anterior pituitary glands of gonadectomized rats that received injections of anti-laminin IgG-HRP were fixed 5 days after injection, lengths of unlabeled basement membrane were distributed between labeled lengths. This indicated that new basement membrane was spliced into old by a process similar to that seen in normal development. Supplementation of gonadectomized rats with testosterone, however, arrested laminin biosynthesis and basement membrane assembly and reversed glandular hypertrophy. These results indicate that, in an absence of sex hormone feedback, renewed synthesis of basement membrane components occurs in the anterior pituitary and is probably necessary to support the additional growth and differentiation of gonadotrophs and other pituitary cells.     

Comparative residual toxicities of pesticides to the predator Agistemus industani (Acari: Stigmaeidae) on citrus in Florida

Residual toxicities of registered and selected experimental pesticides used on citrus against Agistemus industani Gonzalez (Acari: Stigmaeidae) were compared. Pesticides considered highly toxic to A. industani were: abamectin 0.15 EC at 731ml/ha+FC 435-66 petroleum oil at 46.8l/ha, pyridaben 75WP at 469g/ha, ethion 4EC at 7.01l/ha+FC 435-66 petroleum oil at 46.8l/ha, propargite 6.55 EC at 3.51l/ha, chlorfenapyr 2SC at 1.46l/ha applied alone or in combination with FC 435-66 petroleum oil at 46.8l/ha, sulphur 80DF at 16.81kg/ha, dicofol 4EC at 7.01l/ha, fenbutatin oxide 50WP at 2.24kg/ha, benomyl 50WP at 2.24kg/ha, benomyl 50WP at 1.68kg/ha+ferbam 76 GF at 5.60kg/ha, ferbam 76GF at 11.21kg/ha, neem oil 90EC at 46.8l/ha, and copper hydroxide DF (40% metallic copper) at 4.48kg metallic copper/ha+FC 435-66 petroleum oil at 46.8l/ha. Pesticides that were moderately to slightly toxic included: copper sulphate 98% at 4.48kg metallic copper/ha+FC 435-66 petroleum oil at 46.8l/ha, fenbuconazole 2F at 280ml/ha+FC 435-66 petroleum oil at 46.8l/ha, FC 435-66 petroleum oil applied alone at 46.8l/ha or 23.4l/ha, and diflubenzuron 25WP at 1.40kg/ha. Pesticides that were non-toxic included: fenbuconazole 2F at 585ml/ha, malathion 57EC at 5.85l/ha, FC 435-66 petroleum oil at 46.8l/ha, carbaryl 80S at 3.36kg/ha, chlorpyrifos 4EC at 4.68l/ha, and formetanate 92SP at 1.12kg/ha. Understanding the toxic effects of field weathered pesticides against key predacious mite species is important for effective IPM. The results of this study provide a comparison of direct and indirect toxic effects of various pesticides to A. industani under field conditions.     

Primary structure of the hemoglobin beta-chain of rose-ringed parakeet (Psittacula krameri)

The primary structure of Rose-ringed Parakeet hemoglobin -chain was established, completing the analysis of this hemoglobin. Comparisons with other avian -chains show variations smaller than those for the corresponding -chains. There are 11 amino acid exchanges in relationship to the only other characterized psittaciform -chain, and a total of 35 positions are affected by differences among all avian -chains analyzed (versus 61 for the -chains). At three positions, the Psittacula -chain has residues unique to this species. Three ₁₁ contacts are modified, by substitutions at positions 51, 116, and 125.     

Biosynthesis of RNA polymerase in Escherichia coli

Summary In spite of the generally well-coordinated synthesis of RNA polymerase core enzyme subunits (, and ) in Escherichia coli, a situation was found during the growth transition from exponential to stationary phase in which this coordination was broken (the order of differential repression being ; Kawakami et al. (1979)). The present study indicates that, during a certain period of the growth transition, twice as much subunit is synthesized as subunit and the overproduced subunit accumulates as the assembly intermediate ₂ complex, which is rapidly and preferentially degraded.Two independent factors, i.e., carbon source down-shift and oxygen depletion, were examined separately for their influence on the coordinated regulation of the synthesis of RNA polymerase subunits. The depletion of glucose added as a sole carbon source was accompanied by repression of the synthesis of all core enzyme subunits, while under the same conditions the differential rate of subunit synthesis increased. In contrast, the sudden ending of the oxygen supply resulted in specific repression of the synthesis of only and subunits but not of and subunits. The latter result may be explained by the autogenous repression of the rpoBC genes by a temporal increase in the amount of unused cytoplasmic RNA polymerase.Paper XI in this series is Kawakami and Ishihama (1980)     

Unique sequences for two HLA-DRB1 genes expressed on distinct DRw6 haplotypes

We have used restriction fragment length polymorphism (RFLP) analysis and DNA sequencing to characterize two distinct DRB1 alleles expressed on DRw52 and DQw7-associated haplotypes but not readily defined by conventional DR serology. These two haplotypes, designated HLA-D HAG and PEV, react variably with DRw13(w6), DRw14(w6), and the more broad DR 3+6 antisera. Analysis of RFLP revealed that HLA-D HAG and PEV are associated with different DRw52 variants, and that HAG is indistinguishable from DRw18(3) haplotypes. Sequencing of the HAG and PEV DRB1 genes showed each to represent novel alleles. Nevertheless, these sequences show similarities with the other alleles of the DR5, w6, and w8 family. HAG (DRB1*1303) appears to have arisen either from two recombinational events involving at least three DRB1 sequences (DRB1*1101, DRB1*0803, DRB1*0401) or from a single recombinational event together with multiple point mutational events. PEV appears to represent a DRB1*1301-1302/DRB1*1101 recombinant allele, with recombination having occured in the region of bases 175 – 198. The results of this study suggest that the DRw52 family haplotypes is derived from a relatively restricted number of ancestral sequences, with diversity among DRB1 alleles within this family arising through gene conversion or recombination events.     

Action of rat liver Galβ1-4GlcNAc α(2-6)-sialyltransferase on Manβ1-4GlcNAcβ-OMe,GalNAcβ1-4GlcNAcβ-OMe,Glcβ1-4GlcNAcβ-OMe and GlcNAcβ1-4GlcNAcβ-OMe as synthetic substrates

Incubation of synthetic Man\1-4GlcNAc-OMe, GalNAc1-4GlcNAc-OMe, Glc1-4GlcNAc-OMe, and GlcNAc1-4GlcNac-OMe with CMP-Neu5Ac and rat liver Gal1-4GlcNAc (2-6)-sialyltransferase resulted in the formation of Neu5Ac2-6Man1-4GlcNAc-OMe, Neu5Ac2-6GalNAc1-4GlcNAc-OMe, Neu5Ac2-6Glc1-4GlcNAc-OMe and Neu5Ac2-6GlcNAc1-4GlcNAc-OMe, respectively. Under conditions which led to quantitative conversion of Gal1-4GlcNAc-OEt into Neu5Ac2-6Gal1-4GlcNAc-OEt, the aforementioned products were obtained in yields of 4%, 48%, 16% and 8%, respectively. HPLC on Partisil 10 SAX was used to isolate the various sialyltrisaccharides, and identification was carried out using 1- and 2-dimensional 500-MHz¹H-NMR spectroscopy.Abbreviations 2D 2-dimensional - CMP cytidine 5-monophosphate - CMP-Neu5Ac cytidine 5-monophospho--N-acetylneuraminic acid - COSY correlation spectroscopy - DQF double quantum filtered - HOHAHA homonuclear Hartmann-Hahn - MLEV composite pulse devised by M. Levitt - Neu5Ac N-acetylneuraminic acid - Neu5Ac2en 2-deoxy-2,3-didehydro-N-acetylneuraminic acid     

Studies on aneuploids of Petunia

Summary The seven possible primary trisomics of Petunia (2 n= 14) located in the progenies of triploid, hypertriploid and hypotriploid plants were distinguished from one another and from diploid on the basis of cytological and morphological criteria. They were provisionally named as Oval, Semi, Slender, Pseudonormal, Arrow, Narrow and Giant. In three of the trisomics, the extra chromosome was identified for the first time at pachytene stage. Postpachytene studies revealed no precise relationship between the length of extra chromosome and the frequency of multiple association.     

Production of a novel syrup containing neofructo-oligosaccharides by the cells of Penicillium citrinum

A novel syrup containing neofructo-oligosaccharides was produced from sucrose (Brix 70) by whole cells of Penicillium citrinum. The efficiency of fructo-oligosaccharides production was more than 55% and those of the main carbohydrate components, 1-kestose (Fruf 21Fruf 21 Glc), nystose (Fruf 21Fruf 21 Fruf 21 Glc) and neokestose (Fruf 26 Glc12 Fruf), were 22, 14 and 11%, respectively.     

MHC class II sequences of an HLA-DR2 narcoleptic

Narcolepsy has a 98% association with the DR2-Dw2/DQw1 haplotype. To establish if a disease-specific allele is present in narcolepsy, a cDNA library was made from a B-cell line from a DR2,4/DQw1,3 narcoleptic. Clones encoding the two expressed DR2 chains, along with DQw1 and chains, were isolated and completely sequenced. The coding regions of these four genes were similar to published nucleotide and protein sequences from corresponding healthy controls, with some minor exceptions. The 3 untranslated region of one of the DR2 genes in the narcoleptic was extended by 42 bp. Complete sequences were not available for DQw1.2 or from healthy individuals, but first domain nucleotide sequences showed only a single nonproductive difference in DQ. Partial protein sequences of both DQ and from published data were identical. Although the effects of minor differences cannot be ruled out completely, it is concluded that there are probably no narcolepsy-specific DR or DQ / sequences, and that the alleles found in narcolepsy are representative of those found in the healthy population.     

A glutaraldehyde-fuchsin reagent

Synopsis A glutaraldehyde-fuchsin reagent is described. Its preparation avoids the time-consuming ripening period, associated with Gomori's aldehyde-fuchsin. Elastic tissue, insular -cells (after an appropriate prior oxidation) and various other tissue components (e.g. pituitary basophils and acid mucosubstances) can be stained with the reagent 1–2 hours after its preparation.     

Effect of phosphate and adenine nucleotides on the rate of labeling of functional groups at the catalytic site of F1-ATPase

The effect of inorganic phosphate, ADP, ATP, and their analogues on the rate of labeling of F₁-ATPase by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) and phenylglyoxal have been investigated. Analysis of the kinetic data indicate that the labeled functional groups of the essential tyrosine and arginine residues respectively are both located at the catalytic site of F₁. The active phenolic group of tyrosine is located closer to the bound inorganic phosphate or the -phosphate group than the - and -phosphate groups of the bound ATP at the catalytic site, whereas the guanidinium group of arginine is located closer to the - and -phosphate groups of the bound ATP than to its -phosphate group or the bound inorganic phosphate. The kinetically deduced dissociation constants are 1.3 mM and 210 µM for the inorganic phosphate and ADP respectively bound to this catalytic site. Labeling the essential tyrosine residue by NDB-Cl has been found to facilitate subsequent labeling of the essential arginine residue by phenylglyoxal.Abbreviations NBD-Cl 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (this compound has been named 4-chloro-7-nitro-benzofurazan and abbreviated NBf-Cl elsewhere) - DTT dithiothreitol - EDTA ethylenediaminetetraacetic acid - P_i inorganic phosphate - PEP phosphoenolpyruvate - ADPCP ,-methylene-adenosine 5-triphosphate - AMPCP ,-methylene-adenosine 5-diphosphate - Hepes N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - Tris 2-amino-2(hydroxymethyl)-1,3-propanediol     

17 β-Estradiol Enhances the Outgrowth and Survival of Neocortical Neurons in Culture

Results of this investigation demonstrate that exposure to 17 -estradiol differentially and significantly regulates cortical nerve cell outgrowth depending on the cortical region. Parietal and occipital neurons treated with 1 nM 17 -estradiol showed a greater magnitude of neuronal outgrowth whereas outgrowth of temporal cortex neurons was decreased in the presence of 1 nM 17 -estradiol. Frontal cortex neurons showed a consistent enhancement of neuronal outgrowth that did not reach statistical significance. The dose response profile for 17 -estradiol regulation of the macromorphological features exhibited a bimodal dose response relationship whereas the dose response profile for 17 -estradiol regulation of the micromorphological features exhibited a dose response more characteristic of an inverted V-shaped function. An antagonist to the NMDA receptor antagonist, AP5, abolished the growth promoting effect of 17 -estradiol whereas the nuclear estrogen receptor antagonist ICI 182,780 did not. Lastly, neocortical neurons exposed to 17 -estradiol exhibited greater viability and survival than control neurons over a two week period. These data indicate that 17 -estradiol can enhance the growth and viability of select populations of neocortical neurons and that the growth promoting effects of 17 -estradiol can be blocked by an antagonist to the NMDA glutamate receptor and not by an antagonist to the estrogen nuclear receptor.     

The linear tetrasaccharide,Galβ1-4GlcNAcβ1-6Galβ1-4GlcNAc,isolated from radiolabeled teratocarcinoma poly-N-acetyllactosaminoglycan resists the action ofE. freundii endo-β-galactosidase

A novel linear tetrasaccharide, Gal1-4GlcNAc1-6Gal1-4GlcNAc, was isolated from partial acid hydrolysates of metabolically labeled poly-N-acetyllactosaminoglycans of murine teratocarcinoma cells. It was characterized by exo-glycosidase sequencing and by mild acid hydrolysis followed by identification of all partial cleavage products. The tetrasaccharide, and likewise labelled GlcNAc1-6Gal1-4GlcNAc, resisted the action of endo--galactosidase (EC 3.2.1.103) fromE. freundii at a concentration of 125 mU/ml, while the isomeric, radioactive teratocarcinoma saccharides Gal1-4GlcNAc1-3Gal1-4GlcNAc and GlcNAc1-3Gal1-4GlcNAc were cleaved in the expected manner.Abbreviations WGA wheat germ agglutinin - BSA bovine serum albumin - [³H]GlcNAc1-4-GlcNAc1-4GlcNAcOL N,N,NN'-triacetylchitotriose reduced with NaB³H₄     

Changes in the distribution of integrins and their basement membrane ligands during development of human thyroid follicular epithelium

Interactions between cells and basement membrane components are crucial for the regulation of epithelial cell differentiation and polarization. We have studied by immunohistochemical methods the distribution of integrin adhesion proteins and some of their basement membrane ligands in foetal (16--19 weeks) and adult thyroid follicular epithelia. A diffuse immunoreactivity for only 3, v and 1 integrins was found in foetal follicular epithelium, whereas in adult follicular epithelium these integrins were expressed basally in a polarized manner. Additionally, 3 integrin was seen in a more basolaterally confined manner in adult follicular epithelium. Among basement membrane components, laminin 1, 1, 1 and 2 chains were found in epithelial basement membranes of the foetal thyroid gland, suggestive of the presence of laminins-1 and -3. In contrast, the basement membranes of adult follicular epithelium presented a much weaker immunoreactivity for the laminin 2 chain. Furthermore, immunoreactivity for the laminin 2 chain was occasionally seen in adult thyroid glands, apparently confined to myofibroblasts. Immunoreactivity for type IV collagen 1 and 2 (IV) chains was found in follicular basement membranes of foetal as well as adult thyroid gland. The results suggest that during maturation of foetal thyroid follicular epithelium a distinct polarization of integrins takes place. In mature thyroid follicular epithelium, the presumable adhesion-mediating integrin complexes are 31, v1 and/or v3 mediating adhesion to laminin-1 (1-1- 1) and type IV collagen trimer 12 (IV)