Structure determination of the disialylated poly-(N-acetyllactosamine)-containingO-linked carbohydrate chains of equine chorionic gonadotropin

The disialylated poly-(N-acetyllactosamine)-containingO-linked oligosaccharide alditols, released by alkaline borohydride treatment of the enzymicallyN-deglycosylated β-subunit of equine chorionic chonadotropin, were purified by fast protein liquid chromatography (FPLC) on Mono Q and analysed by fast ion bombardment mass spectrometry (FAB-MS) and¹H-NMR spectroscopy. The identified oligosaccharide alditols have the following structure: $$\begin{gathered} Neu5Ac\alpha 2 - 3\left[ {Gal\beta 1 - 4GlcNAc\beta 1 - 3} \right]_{0 - 4} Gal\beta 1 - 4GlcNAc\beta 1 - 6 \hfill \\ \begin{array}{*{20}c} { \backslash } \\ { GalNAc - ol} \\ { /} \\ {Neu5Ac\alpha 2 - 3Gal\beta 1 - 3} \\ \end{array} \hfill \\ \end{gathered}$$      

Simultaneous nonlinear equations of growth

The simultaneous equations

$$\begin{gathered} \frac{{dx}}{{dt}} = \frac{{a_x }}{{k_x }}[k_x - x - f_x (y)] x \hfill \\ \frac{{dy}}{{dt}} = \frac{{a_y }}{{k_y }}[k_y - y - f_y (x)] y \hfill \\ \end{gathered}$$     

Using computer algebra to determine rate constants in biochemistry

In earlier work we have described how computer algebra may be used to derive composite rate laws for complete systems of equations, using the mathematical technique of Gröbner Bases (Bennett, Davenport and Sauro, 1988). Such composite rate laws may then be fitted to experimental data to yield estimates of kinetic parameters. Recently we have been investigating the practical application of this methodology to the estimation of kinetic parameters for the closed two enzyme system of aspartate aminotransferase (AAT) and malate dehydrogenase (MDH) (Fisher 1990a; Fisher 1990b; Bennett and Fisher, 1990): $$\begin{gathered} aspartate + \alpha - ketoglutarate\begin{array}{*{20}c} \rightharpoonup \\ \leftharpoondown \\ \end{array} glutamate + oxaloacetate \hfill \\ {\text{oxaloacetate + NADH}}\begin{array}{*{20}c} \rightharpoonup \\ \leftharpoondown \\ \end{array} malate + NAD^ + \hfill \\ \end{gathered} $$ In this paper we present a fuller (although not yet complete) analysis of the system. We show how symbolic estimates of the error behaviour of the parameters can be made, and used to identify those which are of kinetic significance. Finally we consider how metabolic control analysis can be applied directly to such a system.     

Chemical characterization of the O-specific chain of Sphaerotilus natans ATCC 13338 lipopolysaccharide

The chemical structure of the O-specific chain of the Sphaerotilus natans lipopolysaccharide was established. The isolated polysaccharide moiety contained neutral sugars (rhamnose, glucose, and l-glycero-d-manno-heptose), 3-deoxy-d-manno-octulosonic acid (Kdo), phosphate and ethanolamine. Alditol acetate and methylation analyses showed that the O-specific chain contained linear pentasaccharide repeating units, composed of four units of rhamnose, substituted at 3-position, and one unit of glucose, substituted at 4-position. Oxidation by chromium trioxide showed that all sugars were α-linked. The structure proposed for the O-specific chain has been confirmed by periodate oxidation, and by ¹H-and ¹³C-NMR spectroscopic studies for the original and the Smith-degraded PS moiety. The O-specific unit of the S. natans LPS has the following structure: $$\begin{gathered} [ \to 4) - Glup - (1\xrightarrow{a}3) - Rhap - (1\xrightarrow{a}3) - Rhap - \hfill \\ - (1\xrightarrow{a}3) - Rhap - (1\xrightarrow{a}3) - Rhap - (1]_n \xrightarrow{a} \hfill \\ \end{gathered} $$      

Isolation and1H-NMR spectroscopic identification of the glucose-containing lipid-linked precursor oligosaccharide ofN-linked carbohydrate chains

The lipid-linked precursor ofN-type glycoprotein oligosaccharides was isolated from porcine thyroid microsomes after in cubation with UDP[³H] Glucose. The carbohydrate was released from dolichol pyrophosphate by mild acid hydrolysis, purified by gel filtration and characterized by 500-MHz¹H-NMR spectroscopy in combination with enzymatic degradation. The parent oligosaccharide was found to be Glc₃Man₉Glc-NAc₂. The three glucose residues are present in the linear sequence Glcα1-2Glα1-3 Glc, the latter being α(1-3)-linked to one of the mannose residues. In order to establish the branch location of the triglucosyl unit, the parent compound was digested with jack-bean α-mannosidase. The oligosaccharide product was purified by gel filtration, and identified by¹H-NMR as Glc₃Man₅GlcNAc₂ lacking the mannose residues A, D₂, B and D₃. Therefore, the structure of the precursor oligosaccharide is as follows: $$\begin{gathered} c b a D_1 C 4 \hfill \\ Glc\alpha 1 - 2Glc\alpha 1 - 3Glc\alpha 1 - 3Man\alpha 1 - 2Man\alpha 1 - 2Man\alpha 1 \hfill \\ 3 \swarrow 3 2 1 \hfill \\ Man\alpha 1 - 2Man\alpha 1 Man\beta 1 - 4GlcNAc\beta 1 - 4GlcNAc \hfill \\ D_{2 } A 3 6 \hfill \\ Man\alpha 1 \hfill \\ 6 \hfill \\ Man\alpha 1 - 2Man\alpha 1 \nwarrow 4 \hfill \\ D_3 B \hfill \\ \end{gathered} $$      

Contribution to the genetics of the serum β-lipoproteins in man

A study was made of the genetic behaviour of the factors Ag(x) and Ag(y) of the β-lipoproteins of human serum. It was found that these factors are controlled by a single pair of autosomal codominant genes with complete penetrance at birth. The gene frequencies were:

$$\begin{gathered} Milan . . . . Ag^x = 0,23 Ag^y = 0,77 \hfill \\ Berne . . . . Ag^x = 0,24 Ag^y = 0,76. \hfill \\ \end{gathered}$$     

The basic flow equations of electrophysiology in the presence of chemical reactions: I. Development of the equations

Starting from the basic flux equation, it is possible to obtain an integral form relating the current componentsI _i at an arbitrary pointr ₂ to the distribution of mobilities and concentrationsc _i, potential forces\(\bar \mu \), and chemical productivityp _i without any restrictive assumptions such as constant mobilities, constant field, steady state, or electrical neutrality. The equation is

$$\begin{gathered} I_i (r_2 ) = G_i (r_2 )\left[ {\Delta \bar \mu _i - \int_{r_1 }^{r_2 } {z_i } FA\left( {p_i - dc_i /dt} \right)\left( {\frac{1}{{G_i (r)}}} \right)dr} \right]; \hfill \\ G_i (r) = 1/\int_{r_1 }^r {\frac{{dr}}{{z_i^2 F^2 c_i u_i }}.} \hfill \\ \end{gathered} $$     

Isolation of three novel cholinergic neuron-specific gangliosides from bovine brain and theirin vitro syntheses

In the present study, three extremely minor but novel Chol-1 antigens, termed X1, X2, and X3 have been isolated from bovine brain gangliosides. Based on the results of sialidase degradation, TLC-immunostaining with anti-Chol-1 antibody and fast atom bombardment mass spectrometry, their chemical structures were identified as: $$\begin{gathered} III^6 NeuAc--GgOse4Cer (X1:GM1\alpha ) \hfill \\ III^6 NeuAc,II^3 NeuAc--GgOse4Cer (X2:GT1a\alpha ) \hfill \\ III^6 NeuAc,II^3 NeuAc--NeuGc--GgOse4Cer (X3:GT1b\alpha ) \hfill \\ \end{gathered} $$ The yields of GM1α, GD1aα, and GT1bα, were approximately 150, 20, and 10 µg, respectively, from 10 g of the bovine brain ganglioside mixture. In conjunction with our previous observations, all gangliosides with anti-Chol-1 reactivity were found to contain a common sialyl α2–6N-acetylgalactosamine residue, indicating that this unique sialyl linkage is the specific antigenic determinant. We subsequently examined the biosyntheses of the three novel Chol-1 gangliosides using rat liver Golgi fraction as an enzyme source. The results showed that GM1α, GD1aα, and GT1bα were synthesized from asialo-GM1, GM1a, and GD1b, respectively, by the action of a GalNAc α2-6sialyltransferase.     

A novel pathway of peptide biosynthesis found in methanogenic Archaea

The peptide subunits of the pseudomurein, the cell-wall peptidoglycan of some methanogens, are usually composed of glutamic acid, alanine and lysine. In order to get a more detailed picture of the biosynthetic pathway of the peptide subunit, we performed in vitro assays. Starting from glutamic acid a pentapeptide was obtained in seven steps:

Kinetics and saturation of light-induced near-infrared scattering changes in isolated bovine rod outer segments

The axial and radial shrinkage of bovine rod outer segments, monitored by near-infrared scattering changes (P-signal), is investigated in dependence on the intensity of the activating flash. Suspensions of axially oriented and randomly oriented rod outer segments were measured. In the latter case, axial and radial effects are superimposed to another. The following results are obtained:

1. The axial signal (P _a, Τ≈10 ms) and the radial signal (P _r, Τ=40–100 ms), simultaneously measured on axially oriented rod outer segments, are similarly saturated with a half-saturation at a rhodopsin turnover of 3.5%.
2. For the saturation of the signal amplitude, measured on randomly oriented rod outer segments, a good fit is obtained by: $$\begin{gathered} P\left( \varrho \right) \sim 1 - e\beta \varrho , \hfill \\ \varrho : relative rhodopsin turnover by the flash; \hfill \\ \beta is found in the range 23 \leqslant \beta \leqslant 27 in all measurements \hfill \\ \end{gathered} $$
3. The kinetics of the signal, also measured on the isotropic sample, depends on the rhodopsin turnover, the apparent time constant becoming faster with increasing turnover. The distortion of the signal cannot be fitted by a sum of exponentials with a fixed set of time constants.

The signals from the isotropic sample are fitted by a phenomenological model. It introduces three first order processes concatenated in series; the first step is assumed as a rhodopsin transition inducing the two further processes. The distortion of the signals with increasing? is then described assuming a?-dependent quenching of this induction, according to the measured amplitude saturation. The time constants remain thereby unchanged. The fit yields the values ln 2/k=4, 11, and 45 ms with mean square deviations of 20%.     

Structural analysis of novel rhamnose-branched oligosaccharides from the glycophosphosphingolipids ofLeptomonas samueli

Mild alkaline hydrolysis of the glycophosphosphingolipids of the protozoanLeptomonas samueli liberated several phosphoinositol-containing oligosaccharides (PI-oligosaccharides), which were purified by high performance anion exchange chromatography. The oligosaccharides in the resulting four fractions were characterized by methylation analysis, fast atom bombardment mass spectrometry and two-dimensional nuclear magnetic resonance spectroscopy. The oligosaccharides contain the core structure Man(1–4)GlcN(1–6)-myo-inositol-1-OPO₃, and are substituted with 2mol of 2-aminoethylphosphonate per mol of oligosaccharide. The nonreducing ends of the oligosaccharides were terminated by rhamnose branched neutral and acidic xylose-containing penta-, hexa-, hepta- and octasaccharides, of which the three most abundant were shown to have the structures:

Chemolithotrophic growth of Desulfovibrio sulfodismutans sp. nov. by disproportionation of inorganic sulfur compounds

A new Desulfovibrio strain ThAc01 was isolated from freshwater mud; the strain conserved energy for growth under strictly anaerobic conditions by disproportionation of thiosulfate or sulfite to sulfate and sulfide according to the following reactions: $$\begin{gathered} S_2 O_3^{2 - } + H_2 O \to SO_4^{2 - } + HS^ - + H^ + \hfill \\ 4SO_3^{2 - } + H^ + {\text{ }} \to 3SO_4^{2 - } + HS^ - \hfill \\ \end{gathered}$$ Strain ThAc01 required acetate as a carbon source, but was unable to utilize acetate as an oxidizable energy source. In a defined medium with acetate and bicarbonate as carbon sources, the growth yields per mol of substrate disproportionated were 2.1 g or 3.2 g dry cell mass on thiosulfate or sulfite, respectively. Strain ThAc01 was also able to grow by dissimilatory sulfate reduction with lactate, ethanol, propanol, or butanol as electron donors and carbon sources which were incompletely oxidized to the corresponding fatty acids. However, growth by sulfate reduction was slower than by disproportionation. Elemental sulfur, nitrate, fumarate, or malate did not serve as electron acceptors. Strain ThAc01 contained desulfoviridin and cytochromes; it required panthothenate and biotin as growth factors and had a DNA base ratio of 64.1 mol% G+C. Disproportionating bacteria similar to strain ThAc01 were enriched with either thiosulfate or sulfite from various freshwater, brackish or marine mud samples. Most probable number enumeration indicated that 2×10⁶ thiosulfate-disproportionating bacteria were present per ml freshwater mud. Of various other sulfate-reducing bacteria tested, only Desulfobacter curvatus (strain AcRM3) was able to disproportionate thiosulfate or sulfite. Desulfovibrio vulgaris (strain Marburg) slowly disproportionated sulfite, but effected only a slight increase in cell density. Strain ThAc01 is proposed as the type strain of a new species, Desulfovibrio sulfodismutans.     

A note on the tangle model for DNA recombination

The tangle model developed by Ernst and Sumners provides a rigorous framework to study processive DNA recombination. We suggest here a slight modification of that model. The tangle equations become:

Kinetics of ethanol inhibition in alcohol fermentation

The inhibitory effect of ethanol on yeast growth and fermentation has been studied for the strain Saccharomyces cerevisiae ATCC No. 4126 under anaerobic batch conditions. The results obtained reveal that there is no striking difference between the response of growth and ethanol fermentation. Two kinetic models are also proposed to describe the kinetic pattern of ethanol inhibition on the specific rates of growth and ethanol fermentation: \documentclass{article}\pagestyle{empty}\begin{document}$$\begin{array}{*{20}c} {\frac{{\mu _i }}{{\mu _0 }} = 1{\rm } - {\rm }\left( {\frac{P}{{P_m }}} \right);\alpha } \hfill & {\left( {{\rm for}\ {\rm growth}} \right)} \hfill \\ {\frac{{\nu _i }}{{\nu _0 }} = 1{\rm } - {\rm }\left( {\frac{P}{{P'_m }}} \right);\beta } \hfill & {\left( {{\rm for}\ {\rm ethanol}\ {\rm production}} \right)} \hfill \\ \end{array}$$\end{document} The maximum allowable ethanol concentration above which cells do not grow was predicted to be 112 g/L. The ethanol-producing capability of the cells was completely inhibited at 115 g/L ethanol. The proposed models appear to accurately represent the experimental data obtained in this study and the literature data.     

Endor characterization and D2O exchange in the \begin{array}{*{20}c} {\mathop Z\nolimits^{\begin{array}{*{20}c} + \\ . \\ \end{array} } } \\ \end{array} /{\kern 1pt} \begin{array}{*{20}c} {\mathop D\nolimits^{\begin{array}{*{20}c} + \\ . \\ \end{array} } } \\ \end{array} radical in photosystem II

The early suggestion by Lozier and Butler (Photochem. Photobiol. 17, 133–137 (1973)) that EPR Signal II arises from radicals associated with the water-splitting process in PSII has been confirmed and extended over the intervening years. Recent work has identified the Signal II radicals, \(\begin{array}{*{20}c} {\mathop D\nolimits^{\begin{array}{*{20}c} + \\ . \\ \end{array} } } \\ \end{array}\) and \(\begin{array}{*{20}c} {\mathop Z\nolimits^{\begin{array}{*{20}c} + \\ . \\ \end{array} } } \\ \end{array}\) , with plastosemiquinone cation species. In the experiments presented here we have used ENDOR spectroscopy and D₂O/H₂O exchange to characterize these paramagnets in more detail. The ENDOR matrix region, which arises from protons which interact weakly with the unpaired electron spin, is well-resolved at 4 K and at least seven resonances are apparent. A number of hyperfine couplings in the 3–8 MHz range are observed and are suggested to arise from methyl or hydroxyl protons which occur as substituents on the plastosemiquinone cation ring or from amino acid protons hydrogen-bonded to the 1,4-hydroxyl groups. Orientation selection experiments are consistent with these possibilities. D₂O/H₂O exchange shows that the D⁺/Z⁺ site is accessible to solvent. However, the exchange occurs slowly and is not complete even after 72 hours which suggests that the free radicals are functionally isolated from solvent water.     

Measurement of amount of nitrogen fixed by a legume crop

Summary The amount of nitrogen fixed by a legume crop can be calculated from the relationship % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGceaqabeaacaqGbb% GaaeyBaiaab+gacaqG1bGaaeOBaiaabshacaqGGaGaae4BaiaabAga% caqGGaGaaeOzaiaabMgacaqG4bGaaeyzaiaabsgacaqGGaGaaeOtai% aabccacaqG9aaabaGaaeypaiaabccadaqadaqaaiaaigdacqGHsisl% daWcaaqaaiGacggacaGG0bGaai4Baiaac2gacaGGGaGaaiyjaiaacc% cacaGGobGaaiylaiaacgdacaGG1aGaaiiiaiaacwgacaGG4bGaai4y% aiaacwgacaGGZbGaai4CaiaacccacaGGPbGaaiOBaiaacccacaGGSb% GaaiyzaiaacEgacaGG1bGaaiyBaiaacwgacaGGGaGaai4yaiaackha% caGGVbGaaiiCaaqaaiGacggacaGG0bGaai4Baiaac2gacaGGGaGaai% yjaiaacccacaGGobGaaiylaiaacgdacaGG1aGaaiiiaiaacwgacaGG% 4bGaai4yaiaacwgacaGGZbGaai4CaiaacccacaGGPbGaaiOBaiaacc% cacaqGYbGaaeyzaiaabAgacaqGLbGaaeOCaiaabwgacaqGUbGaae4y% aiaabwgacaGGGaGaai4yaiaackhacaGGVbGaaiiCaaaaaiaawIcaca% GLPaaacaqI4bGaaKiiaiaabshacaqGVbGaaeiDaiaabggacaqGSbGa% aeiiaiaab6eacaqGGaGaaeyAaiaab6gacaqGGaGaaeiBaiaabwgaca% qGNbGaaeyDaiaab2gacaqGLbGaaeiiaiaabogacaqGYbGaae4Baiaa% bchaaaaa!9A78!\[\begin{gathered} {\text{Amount of fixed N = }} \hfill \\ {\text{ = }}\left( {1 - \frac{{\operatorname{atom} \% N - 15 excess in legume crop}}{{\operatorname{atom} \% N - 15 excess in {\text{reference}} crop}}} \right)\user1{x }{\text{total N in legume crop}} \hfill \\ \end{gathered} \]when a suitable reference crop is chosen. The implications and interpretation of this method of measurement are described.     

Présence dans une population congolaise deMus (Leggada) triton Th. de femelles hétérozygotes pour une délétion caractérisée par la suppression du bras court de l'un des chromosomes X métacentriques

A sample of 12Mus (Leggada) triton Th. from the region of Bukavu (Democratic Republic of Congo) contains 5 ♂♂ and 7 ♀♀. 2N=32. All the autosomes are acrocentric. The sex-chromosomes of the ♂ are of the typeX—Y, theX beeing a big submetacentric (I.C.=0,4). Three ♀♀ possess two metacentricX, as expected. By four ♀♀, there is only one typicalX whose partner is acrocentric and as long as the long arm of a normalX. ThisX must have been arisen through the deletion of the short arm and is calledX _ddc. The statistical analysis of the sample is compatible with this pattern:

$$\begin{array}{*{20}c} { \circ \circ } \\ { + + } \\ \end{array} \begin{array}{*{20}c} {X---X = 4/9} \\ {X---X_{dc} = 4/9} \\ {X_{dc} ---X_{dc} = 1/9} \\ \end{array} \begin{array}{*{20}c} { \nearrow \nearrow } \\ { \circ \circ } \\ \end{array} \begin{array}{*{20}c} {X---Y = 2/3} \\ {X_{dc} ---Y = 1/3} \\ \end{array} $$     

Correlation of body temperature and of respiration rate of swine with environmental temperature

Correlation analyses were carried out to determine relation of body temperature and respiration rate of three breeds of swine to the environmental temperature. Coefficients of regression were determined for a prediction equation of the form:

$$\begin{array}{*{20}c} {y = a + b_1 x_1 + b_2 x_2 + b_3 x_3 + b_4 x_4 + b_5 x_5 } \\ {where,y = body temperature} \\ {\begin{array}{*{20}c} {x_1 = respiration rate} \\ {x_2 = body weight} \\ {\begin{array}{*{20}c} {x_3 = sex} \\ {x_4 = environmental temperature} \\ {x_5 = x_1 x_4 } \\ \end{array} } \\ \end{array} } \\ \end{array}$$     

The Tryptophan Fluorescence of Tetracarbidium conophorum Agglutinin II and a Solution-Based Assay for the Binding of a Biantennary Glycopeptide

The plant lectin Tetracarbidium conophorum agglutinin II binds to glycoproteins and glycopeptides in a structurally specific manner [Animashaun et al., (1994) Glycoconjugate J. 11, 299–303]. We have characterized the steady-state and time-resolved fluorescence of the tryptophan residues of this lectin. The fluorescence (λ_ex = 295 nm, λ_em = 350 nm) decay is complex and can be described by four decay times with the following values: τ₁ = 7.4nsec, α₁ = 0.22; τ₂ = 2.9 nsec, α₂ = 0.25; τ₃ = l.0 nsec, α₃ = 0.34; τ₄ = 0.2 nsec, α₄ = 0.18. The addition of a biantennary glycopeptide $\begin{array}{*{20}c} {Gal\beta (1 \to 4)GlcNAc\beta (1 \to 2)Man\alpha (1 \to 6)\neg } \\ {Man\beta (1 \to 4)GlcNAC\beta (1 \to 4)GlcAc\beta (1 \to )\begin{array}{*{20}c} {Glu - Nh_2 } \\ | \\ {Asn} \\ | \\ {COOH} \\ \end{array} } \\ {Gal\beta (1 \to 4)GlcNAc\beta (1 \to 2)Man\alpha (1 \to 3)} \\ \end{array} $ to the lectin results in a quench and an 8 nm blue shift of the emission spectrum. The effect is saturable, and is described by an association constant of 1.8×10⁵ M^?1. The tryptophan fluorescence of Tetracarbidium conophorum agglutinin II may therefore be utilized to characterize thermodynamically the binding interactions between this lectin and complex glycoprotein.     

Dynamical systems under constant organization I. Topological analysis of a family of non-linear differential equations —A model for catalytic hypercycles

The paper presents a qualitative analysis of the following systems ofn differential equations: \(\dot x_i = x_i x_j - x_i \sum\nolimits_r^n { = 1} x_r x_s {\mathbf{ }}(j = i - 1 + n\delta _{i1} {\mathbf{ }}and{\mathbf{ }}s = r - 1 + n\delta _{r1} )\) , which show cyclic symmetry. These dynamical systems are of particular interest in the theory of selforganization and biological evolution as well as for application to other fields.