Carcinogenic Helicobacter pylori Strains Selectively Dysregulate the In Vivo Gastric Proteome,Which May Be Associated with Stomach Cancer Progression,

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Highlights

• •H. pylori dysregulates the in vivo gastric proteome of gerbils in a strain-specific manner.
• •H. pylori increases RABEP2 and G3BP2 levels in cell culture.
• •H. pylori upregulates RABEP2 and G3BP2 in gerbil and human gastric epithelium.
• •Levels of RABEP2 and G3BP2 increase with severity of malignant lesions in vivo.

     

Chromatin proteins of rat preputial-gland: Acute changes in response to estrogen

• 1.1. The effects of estradiol-17β (E₂β) at 2 or 15min in vivo on chromatin proteins of rat preputial-gland were analyzed by a battery of electrophoretic methods.
• 2.2. Among histones, E₂β/control ratios for major bands of H1 decreased substantially between 2 and 15 min. In contrast, ratios of H4 increased (P < 0.01), whereas, except for losses by 2 min m a H2B-like component and in H3.1, other core histones were unchanged.
• 3.3. Among 0.35 M NaCl-soluble proteins, components at 34K-mol. wt and < 21K-mol wt were increased after 2 min of E₂β. The bulk of the hormone-responsive low-molecular weight proteins was basic in charge.
• 4.4. Electrophoretic correlates of 6 basic lysosomal proteins corresponded to those of low-molecular weight salt-soluble chromatin proteins.
• 5.5. Selective proteolysis initiated in vivo by E₂β depleted some tightly-bound nonhistone proteins.

     

Histone variants in diptera during metamorphosis

• 1.1. Patterns of histone variants from three diptera, Ceratitis capitata, Dacus oleae and Drosophila melanogaster were compared to mouse and to Plodia interpunctella variant patterns.
• 2.2. The three diptera contain histones which comigrate on two dimensional gels with H3.2, H3.3 and H4 in mouse and Plodia. H2A.1 and H2B.1 comigrate with Plodia H2A.1 and H2B.1 and are different from mouse, whilst H2A.Z has a different mobility to that of Plodia and mouse.
• 3.3. The iodinated peptides obtained from H2A.1 and H2A.2 of the diptera studied are compared to mouse and Plodia H2A.1 and H2A.Z peptides.
• 4.4. The histone variants from three developmental stages, larval, pupal and adult of the three diptera were identified and compared.
• 5.5. The same histone variant pattern is found through all stages of development.

     

Equilibria and kinetic of oxygen and carbon monoxide binding to the haemoglobin of the South American lungfish,Lepidosiren paradoxa

• 1. The haemoglobin of the South American lungfishLepidosiren paradoxa has a single component.
• 2. The equilibria of this respiratory protein with oxygen have been investigated both in the blood and with the purified haemoglobin. There is a substantial, normal, alkaline Bohr effect and marked sensitivity to organic phosphates in the haemoglobin solutions.
• 3. Studies on the pH dependence of the kinetics of oxygen dissociation can be interpreted in terms of a normal Bohr effect.
• 4. The kinetics of combination of carbon monoxide have an unusual pH dependence.
• 5. These findings are discussed in terms of the two-state model of Monodet al. (1965)

     

Effect of pH on the kinetics of oxygen and carbon monoxide reactions with hemoglobin from the air-breathing fish,Loricariichthys

• 1.Loricariichthys sp., an air-breathing fish from the Amazon River has one major hemoglobin component.
• 2. Quantitative studies on the kinetics of O₂ dissociation and CO combination to the protein were performed by stopped-flow experiments at different pH values and a constant ATP concentration of 1.25 mM.
• 3. The oxygen dissociation shows a simple first order behavior and a strong pH dependence.
• 4. The CO combination kinetics, on the other hand, were homogeneous and fast at higher pH values and slow and clearly autocatalytic at pH values below 7.0.

     

Purification and characterization of the protamines and related proteins from the sperm of a tunicate,Styela plicata

• 1.1. We have characterized for the first time the major sperm-specific nuclear proteins (X, P1 and P2) of the tunicate Styela plicata. Both P1 and P2 have an amino acid composition that allows us to classify them as protamine-like proteins.
• 2.2. The protein P1 of lower electrophoretic mobility has a trypsin-resistant core which is compositionally related to that of histones of the H1 family and to the PL-I protein found in the sperm of marine invertebrates. The evolutionary significance of this finding is discussed.
• 3.3. In addition to P1 and P2, the sperm nucleus of S. plicata contains a protein X component which is also compositionally related to PL-I proteins from bivalve molluscs.
• 4.4. Besides these sperm-specific proteins, a full complement of somatic-like histones, including a somatic-like histone H1, is also present. These histones represent only a small fraction of the total nuclear proteins of the sperm.

     

Alkylating spin labels for spin-labelling of DNA

• 1.1. Five alkylating spin labels, termed SL1, SL2, SL3, SL4 and SL5, have been synthesized and their suitability for spin-labelling of DNA was studied.
• 2.2. It was found that one of them (SL5) lost its free radical moiety under the conditions of spin-labelling, thus giving DNA with no EPR signal.
• 3.3. Two of the spin labels (SL2 and SL4) denatured DNA and therefore they were not appropriate for spin-labelling of double stranded DNA.
• 4.4. The spin labels SL1 and SL3 as well as the recently synthesi/ed HMSL [Raikova et al. (1982) Int. J. Biochem. 14, 41–46.] showed a low destabilizing effect on the DNA double helix and gave spin-labelled DNA with very immobilized EPR spectra (2Azz = 40G).
• 5.5. These three spin labels could be recommended as suitable for application in the EPR spectrometry of DNA.

     

Effect of conjugation on the incorporation of glycine into Tetrahymena

• 1.1. Conjugation of Tetrahymena enhanced the incorporation of glycine into the nuclear fraction by 500%.
• 2.2. Incorporation of glycine into the microsomal supernatant was augmented by almost 500% by conjugation.
• 3.3. Mitochondrial incorporation was stimulated nearly 3-fold in the conjugating strains while the incorporation of glycine into the microsomes was enhanced approximately 2.5 times.
• 4.4. In the whole cell, glycine incorporation was increased nearly 2-fold by conjugation.
• 5.5. Strong nuclear involvement was indicated by elevated metabolic activity and incorporation of glycine into RNA and DNA.
• 6.6. Stimulation of the metabolism of Tetrahymena by cell communication suggests that the contents of a cell can have a synergistic effect on another cell.
• 7.7. Augmentation of the biosynthetic capacities of cells by fusion is a demonstration of the dominant role of the cell membrane in the regulation and control of cells.
• 8.8. Enhancement of biosynthesis of nuclear proteins in conjugating strains of cells indicates that fusion gives rise to the synthesis of new protein from previously existing protein or protein procursors.
• 9.9. The specific activities of the subcellular fractions after the incorporation of glycine into 2 separated starved strains of Tetrahymena followed the usual pattern of nucleus less than whole cells, whole cells less than mitochondria, mitochondria less than microsomes, but with the microsomal supernatant being much greater than that of the microsomes.

     

A fetal-maternal shift in the oxygen equilibrium of hemoglobin from the viviparous caecilian,Typhlonectes compressicauda

• 1. The equilibria and kinetics of oxygen binding by blood and hemoglobin from adult and fetal caecilians,Typhlonectes compressicauda, have been measured.
• 2. The oxygen affinity of fetal blood is higher than that of adult blood.
• 3. Electrophoresis of adult and fetal hemoglobins suggests that they may be identical: a major and minor component occurs in each.
• 4. Adult and fetal hemoglobins have identical oxygen equilibria. Stripped hemoglobins have a high oxygen affinity and no Bohr effect between pH 6.5 and 10.0. An “acid”, reversed Bohr effect is present below pH 6.5. The addition of 1 mM ATP reduces the oxygen affinity markedly and produces a moderate, normal Bohr effect.
• 5. The major nucleoside triphosphate in fetal and adult erythrocytes is adenosine triphosphate: about 10% of the nucleoside triphosphates is guanosine triphosphate. Adult erythrocytes contain 3 times as much ATP as do the fetal erythrocytes.
• 6. The fetal to maternal shift in the oxygen equilibrium is mediated entirely by the difference in ATP content of the maternal and fetal red blood cells.

     

Comparative study of hemoglobins from different Artemia populations. The influence of temperature on the oxygen equilibria

• 1.1. The P50 values of extracellular hemoglobin (Hb) of five Artemia populations from different geographical origin are affected by temperature.
• 2.2. The free oxygen binding energy is high for all the populations (ΔH between −34.7 and −56.2kj/mol).
• 3.3. A possible correlation between thermal sensitivity of Hb and the ambient temperature of the habitat must be considered very carefully.
• 4.4. The occurence of different quantities of Hb1 (αα chains) Hb2 (αβ chains) and Hb3 (ββ chains) in the different populations possibly influences thermal sensitivity.

     

Dna breakage and inactivation resulting from hydroxylamine and/or cis-Diamminedichloroplatinum(Ii) interactions with plasmid dna

• 1.1. Occurrence of lesions induced in plasmid DNA by cis-DDP and by HA was quantified both as a transforming activity and as conformation integrity of supercoilcd pBR322 DNA. Fifty per cent decrease of the biological activity of plasmid DNA, not accompanied by measurable change of DNA conformation, was observed after a single exposure of DNA to cis-DDP (1 hr/37°C).
• 2.2. HA induced conversion of supercoiled DNA to other topological forms in a dose-dependent manner.
• 3.3. One- and two-strand DNA breaks were determined electrophoretically with high sensitivity. Cis-DDP exposed DNA relaxed at 30 times lower HA concentration compared to intact DNA.
• 4.4. This effect may be connected with a local distortion of DNA structure at the cis-DDP—DNA bond, which makes possible high effectivity of HA-DNA interaction.
• 5.5. On other hand, biological activity stayed at the 50% level despite breaks induced in DNA.
• 6.6. This finding supports the idea that DNA breaks occur at the locations which were modified during the exposure of DNA to cis-DDP.
• 7.7. The importance of the DNA structure during interaction with HA may be seen during HA-DNA interaction at heat-denaturation of supercoiled DNA. At this condition, the DNA breaks were induced at 100 times lower concentration of HA.
• 8.8. We conclude, on the basis of these results and results published earlier, that local distortion of supercoiled DNA structure, which is caused by the cu-DDP bond, and the local DNA uncoiling caused by heat-denaturation are related to high HA-DNA reactivity.

     

Surface Loops in a Single SH2 Domain Are Capable of Encoding the Spectrum of Specificity of the SH2 Family,

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Highlights

• •Surface loops play an essential role in SH2 domain specificity.
• •Diverse specificities may be obtained from a single SH2 domain by combinatorial mutations in the EF and BG loops.
• •The specificity of a loop mutant correlates with the sequence characteristics of the bait peptide used in its isolation.

     

Isolation of dna and rna from Streptococcus sobrinus OMZ176 using CsTFA gradients

• 1.1. A simple procedure for isolation of high molecular weight genomic DNA, and RNA, from Streptococcus sobrinus OMZ176 is described.
• 2.2. Cell cultures were grown aerobically for 10 hr.
• 3.3. Spheroplast formation and lysis was achieved by mutanolysin/lysozyme-dependent digestion of the cell wall, followed by N-lauroylsarcosinate-mediated lysis.
• 4.4. Nucleic acids were isolated directly from cell-lysates using cesium-trifluoroacetate (CsTFA) densitygradient centrifugation.
• 5.5. Three different centrifugation regimes were tested: self-generated density gradients in a fixed angle rotor; self-generated density-gradients in a swinging-bucket rotor; pre-formed density-gradients in a swinging-bucket rotor.
• 6.6. Genomic DNA isolated by the CsTFA-procedure was found to have higher purity as compared to genomic DNA isolated in a conventional CsCl gradient.
• 7.7. Isolated DNA was shown to be of a quality suitable for applications in molecular biology.

     

Fluorescence studies on the interaction of muscle M-line proteins,creatine kinase and the 165,000 dalton component,with each other and with myosin and myosin subfragments

• 1.1. The binding of 8-anilino-l-naphthalene sulfonate (ANS) by M-line creatine kinase (CPK) and the 165,000 dalton protein component was studied by fluorescence.
• 2.2. One mole of ANS binds to a mole of each M-protein and the binding site on these two M-proteins is hydrophobic in nature.
• 3.3. M-line proteins labeled with ANS were used to demonstrate their interaction with myosin and myosin subfragments 1 and 2.
• 4.4. A unique finding of this study, that labeled M-line CPK binds to subfragment 2 of myosin, is of significance from a structural view point, since only the rod portions of myosin molecules are exposed at the M-line and therefore able to interact with CPK.
• 5.5. The use of a sulfhydryl fluorescent probe, N-(2-iodoacetyl)-N-(5-sulfo-l-napthyl) ethylene diamine, (1,5-AEDANS), has shown that the essential sulfhydryl group on CPK is very important for its interaction with both myosin and the 165,000 dalton M-line protein component.

     

Age-related changes in total dna and rna and incorporation of uridine and thymidine in rat liver,kidney and spleen

• 1.1. Total content of DNA and RNA in liver, kidney and spleen were measured in young and aged rats. At the same time the incorporation of [¹⁴C]thymidine, a DNA precursor, and [³H]uridine, an RNA precursor, were also determined.
• 2.2. Changes in total organ DNA and RNA correlated with sexual maturation as did incorporation of precursors.
• 3.3. Young animals have more DNA per organ relative to RNA. with kidney and spleen DNA showing a decrease between maturity and senescence.
• 4.4. However, liver RNA increases with age. a change probably due to decreased catabolism of RNA since [³H]uridine uptake decreases.
• 5.5. Liver polyploid differentiation, and [¹⁴C]thymidine and [³H]uridine uptake, are correlated.
• 6.6. In kidney, incorporation of [³H]uridine is inversely related to [¹⁴C]thymidine incorporation.

     

The restriction analysis of nDNA from the corvids (Passeriformes,aves)

• 1.1. The nDNA of carrion crow Corvus corone L., hooded crow C. cornix L., their hybrids, as well as magpie Pica pica L., were digested by the tetranucleotide recognizing restriction enzymes Sau3a, AluI, BspRI and then analysed using electrophoresis with microdensitometry.
• 2.2. The distribution patterns of restriction DNA fragments proved to be nuclease- and taxon specific.
• 3.3. The observed families of repeated sequences are characterized by different length (from 30 bp to 23 tbp), number of copies in genome (approximately 10³ and 10⁶) and supposedly different types of organization and evolutionary age.
• 4.4. The total DNA amount identified in the form of discrete fragments is 16 and 19–21% for magpie and crows, respectively.
• 5.5. The DNA restriction patterns of hybrid forms do not differ from the parental species.

     

Purification and characterization of the endonuclease present in Physalia physalis venom

• 1.1. Physalia physalis nematocyst venom contains a DNase which has a non-specific endonucleolytic action.
• 2.2. This enzyme has an approximate molecular weight of 75,000 daltons.
• 3.3. The enzyme can cleave DNA over a wide pH range with an optimum near neutrality.
• 4.4. The enzyme is thermolabile and its activity can be stimulated by 80 nM NaCl or 10 mM MgCl₂.

     

Respiration in the eastern water dragon,Physignathus lesueurii (agamidae)

• 1.1. Some aspects of the gas exchange system of a diving lizard, Physignathus lesuewii were studied.
• 2.2. Breathing patterns were analysed.
• 3.3. Breathing rate increases logarithmically with temperature and Q₁₀ = 1.8. LogBR = −0.237 + 0.0256 T.
• 4.4. Gas tensions in lung air and arterial and venous blood were measured. Arterial pH declines with increasing temperature.
• 5.5. Temperature has a marked effect on oxygen affinity of the blood (ΔH = −10.1 kcal mol⁻). A Bohr effect was also noted.
• 6.6. CO₂ equilibrium curves were drawn.
• 7.7. The results are considered with a view to anticipating the efficiency of the gas exchange system of this species under conditions of variable temperature and during diving.

     

Isolation and characterization of the plasma membrane from slime-forming,encapsulated Streptococcus cremoris

• 1.1. The plasma membrane of slime-forming, encapsulated Streptococcus cremoris from “viili” was isolated in hypotonie conditions in the presence of lysozyme (EC 3.2.1.17) using density gradient centrifugation as the last purification step.
• 2.2. The membrane yield was 15.8% of wet weight cells and the preparation contained 64.4% protein. 19.1% carbohydrate, 5.8% aminosugars, 5.1% RNA and 0.07% DNA.
• 3.3. Buffered 1% (w/v) Triton X-100 solubilized 33.6% of membrane proteins. The number of polypeptides detected by SDS-polyacrylamide gel electrophoresis was 59 when the membrane was isolated without a protease inhibitor and 44 in the presence of a protease inhibitor.
• 4.4. The molecular weights of the polypeptides varied from 13,500 to 100,000.
• 5.5. Ultrathin-layer electrofocusing analysis revealed the range of protein pi values to be between 3.50 and 5.85 concerning 77.3% of proteins and between pI 5.85 and 8.15 concerning 18.2% of proteins.
• 6.6. The isoelectric point of the only basic protein component was 9.3.