Cytogenetics of the parthenogenetic grasshopper Warramaba (formerly Moraba) virgo and its bisexual relatives

The distribution of late-replicating segments along the chromosomes of five clones of W. virgo is described. Some, but not all of these segments correspond to C-bands. In general, the autoradiographic profiles (histograms of linear grain density along the length of chromosomes labeled with tritiated thymidine in late S-phase) show strong resemblances throughout the five clones. However, some significant differences exist, and these are particularly marked in the case of the Boulder clone, which is anomalous in many other respects. — A similar study has also been carried out on the two bisexual species of Warramaba (P169 and P196) that gave rise, by hybridization more than half a million years ago, to the parthenogenetic W. virgo. In the case of P169, the autoradiographic profiles of the three large chromosomes (X+A, B+5, CD) which it has contributed to the W. virgo karyotype are extremely similar to those of the corresponding chromosomes in the virgo clones we have studied. In the case of P196 there is likewise, in most instances, a close resemblance of the autoradiographic profiles of the AB, X₁ and CD chromosomes to those of the same chromosomes in the virgo clones, but that of the X₁ shows no particular resemblance to the anomalous profile of the X₁ in the Boulder clone, in which the X₁ has undergone a structural reorganisation. The autoradiographic profile of the P196 CD chromosome does, however, show a much closer resemblance to that of the corresponding chromosome in the Boulder clone than to those of the CD₁₉₆ in the other four virgo clones studied. These investigations confirm the considerable evolutionary stability of DNA replication patterns.     

Cytogenetics of the parthenogenetic grasshopper Warramaba (formerly Moraba) virgo and its bisexual relatives

Triploid hybrids have been obtained by crossing individuals of the diploid Warramaba virgo with males of two undescribed related species of Warramaba, P169 (neo-XY) and P196 (X₁X₂Y). In both cases, offspring which receive a Y-chromosome from the father are males, while those that receive a neo-X from P169 or an X₁ and an X₂ from P196 are females. The triploids can be distinguished from diploids, even in the earliest nymphal instars, by the larger size of their eye facets. Their gonads are undeveloped and abnormal so that they are mostly sterile (the males absolutely so). Nevertheless, in the case of female hybrids (both the ones between virgo and P169 and those between virgo and P196) a few oocytes do develop and it was possible to obtain a further generation of hybrids by parthenogenesis. These, which are all female, and have karyotypes identical to those of their mothers, are derived from eggs which have undergone the virgo type of meiosis, with a premeiotic doubling of the chromosome number, followed by synapsis restricted to sister chromosomes. — Some diploid hybrids have also been obtained between the bisexual species P169 () and P196 (). In this case the male offspring died in the embryonic stage or immediately after hatching. Female hybrids, on the other hand, were viable but had under-developed ovaries, so that they only laid very few eggs. Some of the latter developed into embryos with a karyotype identical to that of the mother, but the meiosis of these eggs has not yet been studied, so that it is not known whether it is of the virgo type. These hybridization experiments support the hypothesis that virgo originated as a hybrid between P196 and P169. — A single male hybrid between Warramaba picta () and P169 () was obtained; it had active spermatogenesis, but many meiotic abnormalities.     

Cytogenetics of the parthenogenetic grasshopper Warramaba (formerly Moraba) virgo and its bisexual relatives

The karyotypes of Warramaba virgo and the two bisexual species which gave rise to it, by hybridization, have been studied by the C- and G-banding techniques. W. virgo consists of a number of clones which differ by a limited number of inversions, translocations and fusions and also in C-banding pattern. In general, each local population consists of a single clone, although instances are known where two clones exist at the same locality. A total of fifteen cytological clones are here recognised and described. Some of these are known from single localities, while others have been found at several collecting sites in the same general area. The differences between them in banding pattern are relatively minor, i.e. the architecture of the karyotype has been highly conserved. A total of 50 C-bands have been recognised in W. virgo, but some of these are only present in a single clone and most clones have 39–40 bands. Most of these are centromeric or interstitial; a few are terminal (telomeric) in the short arms. There are three types of bands, dark-staining centromeric ones, intermediate ones that are somewhat less densely staining, and lighter ones that appear grey even after successful C-banding. The C-banding pattern of W. virgo provides additional evidence for its origin by hybridization between the bisexual species P196 and P169. It is possible that W. virgo arose only once (i.e. from one or more matings between P196 and P169 individuals in the same deme, within a few generations). However, two clones (Zanthus and Boulder), which are themselves closely similar, differ so much in banding pattern from all others that they may have arisen independently.     

Nucleic acid hybridization of group N and group D streptococci

Streptococci of serological groups N and D were found to belong to three different 23S ribosomal RNA (rRNA) homology clusters which are not closely related to each other. One cluster includes the group N streptococci:Streptococcus lactis, S. cremoris, S. diacetylactis, and, in addition, two lactobacilli, Lactobacillus hordniae andL. xylosus. The second one is composed ofStreptococcus faecium, S. durans, andS. faecalis with its subspecies. All are members of serological group D. The third group includes the group D streptococciStreptococcus bovis andS. equinus together withS. thermophilus andS. salivarius. DNA-DNA hybridization studies confirm the close genetic relationship within each of the rRNA homology clusters.     

Characteristics of ribonucleic acids isolated from Botryodiplodia theobromae pycnidiospores

Nucleic acids isolated from dormant and germinated Botryodiplodia theobromae pycnidiospores contain five distinct species of RNA. They include two ribosomal species, two ribosomal-associated species and transfer RNAs. Sedimentation coefficients of 25.1S and 18S were obtained for the two ribosomal RNA species and 5.8S and 5S for the two ribosomal-associated RNA components. Molecular weights of 1.20, 0.67, 0.054 and 0.035x10⁶ daltons were obtained after formaldehyde treatment and electrophoresis on polyacrylamide gels for these same four RNAs. Methylated nucleotides were present in the transfer RNAs and large and small ribosomal RNAs; in contrast 5.8S and 5S RNAs contained few methylated nucleotides. In addition to the 5 distinct RNA species, polyadenylate-containing RNA was isolated from both dormant and germinated spores.Published with the approval of the Director as paper no. 5006, Journal Series, Nebraska Agricultural Experiment Station. The work was conducted under Nebraska Agricultural Experiment Station Project no. 21-17.     

Holling's “hungry mantid” model for the invertebrate functional response considered as a Markov process. Part II: Negligible handling time

In this paper we analyse a stochastic model for invertebrate predation taking account of the predator's satiation. This model approximates Holling's hungry mantid model when handling time is negligible (see Part I). For this model we derive equations from which we can calculate the functional response and the variance of the total catch. Moreover we study a number of approximations which can be used to calculate these quantities in practical cases in a relatively simple manner.List of Notation a rate constant of digestion - b maximum of rate constant of prey encounter in the mantid - c satiation threshold for search - c satiation threshold for pursuit in the mantid - c _i (w^1/2(N- N)ⁱ) - expectation operator - f rate of change of satiation during search - F functional response: mean number of prey eaten per unit of time - g rate constant of prey capture - h probability generating function of N conditional on S = s times p - H probability generating function of N - m_i ¹ - n, N number of prey caught - p probability density of S - p_n simultaneous probability (density) of N and S - q probability of strike success - r dummy variable in generating function - s, S satiation - T _s search time - T _d digestion time - v asymptotic rate of increase of var v - V asymptotic rate of increase of var N - w weight of edible part of prey - W standard Wiener process - x prey density - z (N{S = s}-N)p - rate constant of prey escape time maximum pursuit time - (v{S = + w ^1/2}-v) - present time as a fraction of the time from the start to the end of the experiment - hazard rate of T _s - mean time between (downward) passages of S through c - v w_–1/2(N-) - edible prey biomass density - probability density of , number pi - parameter of Weibull distribution of T _s = (1/2acx(-g(c)))_1/2 - w_–1/2(S -) - satiation in the guzzler approximation: solution to d/dt = f() + g(), (0)=S(0). - biomass functional response: wF - total biomass catch in the guzzler approximation: solution to d/dt = g(), (0) = 0     

A gene encoding Achlya bisexualis β-amylase and its expression in Saccharomyces cerevisiae

Part of a -amylase genomic DNA sequence from the oomycete, Achlya bisexualis was cloned by polymerase chain reaction (PCR) using degenerate oligonucleotide primers derived from the conserved regions of other known -amylase sequences. The 5- and 3-regions of the -amylase gene were amplified by genome walking method. The Ach. bisexualis -amylase gene consisted of a 1338bp open reading frame, encoding a protein of 446 amino acids with a molecular weight of 49 381Da, and was not interrupted by any intron. The deduced amino acid sequence of the -amylase gene had 67% similarity to the -amylase of Saprolegnia ferax, followed by 40% similarity to that ofArabidopsis thaliana. The -amylase gene was expressed in Saccharomyces cerevisiae placing it under the control of the alcohol dehydrogenase gene (ADC1) promoter.     

Role of mitochondria in ethanol tolerance of Saccharomyces cerevisiae

The presence of active mitochondria and oxidative metabolism is shown to be essential to maintain low inhibition levels by ethanol of the growth rate (), fermentation rate (v) or respiration rate () of Saccharomyces cerevisiae wild type strain S288C. Cells which have respiratory metabolism show K _i (ethanol inhibition constant) values for , v and , higher (K _i>1 M) than those of petite mutants or grande strains grown in anaerobiosis (K _i=0.7 M). In addition, the relationship between or v and ethanol concentration is linear in cells with respiratory metabolism and exponential in cells lacking respiration. When functional mitochondria are transferred to petite mutants, the resulting strain shows K _i values similar to those of the grande strain and the inhibition of and v by increasing ethanol concentrations becomes linear.     

<Emphasis Type="Italic">Tuamotuichthys bispinosus</Emphasis>, a new genus and species from off Tuamotu Islands,South Pacific Ocean (Ophiidiformes,Bythitidae)

A new bythitid genus and species, Tuamotuichthys bispinosus, is described from an adult male (88mm SL), caught by dredge in 1990, off Morane Atoll, Tuamotu Archipelago (23°0843 S, 137°0919 W, 536m). It is characterized by having large scales on head and body, two opercular spines, few gill rakers (total 11, 3 of them long), depressed head, vertically expanded posterior maxilla, and a short predorsal (29% SL), each of which resembles several known genera (Calamopteryx, Cataetyx, Diplacanthopoma, Grammonus, and Lucifuga). The combination of character states, however, does not fit any of the known bythitid genera.     

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.     

Heterochromatin variation in Cryptobothrus chrysophorus

The endemic grasshopper Cryptobothrus chrysophorus is widely distributed throughout S.E. Australia and its populations display an extensive and spectacular pattern of autosomal variation. While the standard telocentric complement of three long (L1–3), six medium (M4–9) and two short (S10–11) autosome pairs is present throughout most of its range, two quite distinct chromosome races can be defined within this species. Populations in the northern part of its distribution (northern N.S.W. and southern Queensland-northern race) are differentiated from the remainder (southern race) by fixed blocks of distal heterochromatin on autosomes M4, 5, 6, 8 and 9 and by differences in the character of the megameric M7 chromosome. Additionally, while many populations in both races show a polymorphic system of supernumerary segments on the two smallest autosomes (S10–11), that found in the northern race is both more variable and more complex. On the other hand all the populations of the southern race we have examined are polymorphic for a series of centric shifts which convert telocentrics into acro- or meta-centrics. These occur more commonly in the megameric M7 and the two smallest autosomes (S10–11) although in one population (Forbes Creek, N.S.W.) at least 12 different shifts involving 8 of the autosomes (L3, M4, 5, 6, 7, 8, 9 and S10) are known. By contrast, in the northern erace only the small autosomes (S10–11) show centric shifts. These several floating and fixed variants thus involve all chromosomes of the standard set other than the two largest autosomes (L1–2) and the X-chromosome, which appear to be invariate. Finally, morphologically distinct supernumerary (B) chromosomes, intermediate in size between the standard S10 and the M9 elements, are found in both races but are especially common in Tasmania, the most southerly point of the species range. These B-chromosomes are partly heterochromatic and partly euchromatic so that they too add to the considerable heterochromatin variation in this species.     

Electrophoretic variation between class II molecules expressed on HLA-DRw8 homozygous typing cells reveals multiple distinct haplotypes

Two-dimensional (2D) gel electrophoresis of immunoprecipitated HLA-DR antigens from eight homozygous typing cells (HTC) expressing the HLA-DRw8 specificity revealed a clustering of polymorphic chain patterns into distinct electrophoretic variants. The variant patterns correlate with three discrete HLA-D clusters that are defined in the mixed leukocyte culture reaction (MLR) using DRw8-positive HTC. These HLA-D clusters have been provisionally designated Dw8.1, detected primarily in Caucasoids, Dw8.2, detected primarily in American Indians, and Dw8.3, detected predominantly in Orientals. All three HLA-Dw8.1 cell lines express a single DR-locus product as defined by immunoprecipitation with a DR-specific monoclonal antibody, P4.1. This DR chain is identical among the Dw8.1 cell lines and different from the DR chains of the Dw8.2 and Dw8.3 cell lines. Two separate Dw8.2 HTC express a shared DR chain that is slightly more basic than the 8.1 DR molecule; interestingly, one of these lines also expresses an additional DR-like chain not found in the other cells. Thus, the two lines defining the Dw8.2 cluster share one distinct class 11 molecule, but differ in another and therefore are not biochemically HLA-identical. Cells from the Dw8.3 cluster are likewise distinct from all other Dw8 clusters. One additional DRw8-positive HTC has been analyzed and found to be distinct from the Dw8.1, 8.2 and 8.3 clusters by both MLR and 2D gels. lmmunoprecipitates using monoclonal antibody 1B5 [anti-DR and anti-DQ(DS)] identify additional polymorphic class II variants among the cell lines tested. These data indicate that HLA-DRw8 is a public serologic specificity present on class II molecules expressed on multiple distinct haplotypes. These haplotypes differ from each other in expression of polymorphic class II molecules encoded by at least two HLA loci. They also differ in HLA-D, even though they all type as HLA-DRw8 homozygous. In Dw8.2, variation in expressed chains is not reflected in variation in HLA-D, indicating that MLR, as well as serologic typing, does not detect the full degree of allelic polymorphism within HLA.     

Conformational Change of the Rieske [2Fe–2S] Protein inCytochrome bc 1 Complex

Structures of mitochondrial bc ₁ complex have been reported based on four different crystalforms by three different groups. In these structures, the extrinsic domain of the Rieske [2Fe–2S]protein, surprisingly, appeared at three different positions: the c ₁ position, where the [2Fe–2S]cluster exists in close proximity to the heme c ₁; the b position, where the [2Fe–2S] clusterexist in close proximity to the cytochrome b; and the intermediate position where the[2Fe–2S] cluster exists in between c ₁ and b positions. The conformational changes betweenthese three positions can be explained by a combination of two rotations; (1) a rotation of theentire extrinsic domain and (2) a relative rotation between the cluster-binding fold and thebase fold within the extrinsic domain. The hydroquinone oxidation and the electron bifurcationmechanism at the Q_P binding pocket of the bc ₁ complex is well explained using theseconformational changes of the Rieske [2Fe–2S] protein.     

Adoptions expérimentales de larves entre des fourmis de genres différents:Leptothorax nylanderi Förster etSolenopsis fugax Latreille

Résumé En l'absence de son propre couvain,Solenopsis fugax a élevé des larves deLeptothorax nylanderi, à la température de 22°C. Les ouvrières deSolenopsis détruisirent une partie de ces larves mais nourrirent celles qu'elles épargnèrent; ces dernières grossirent lentement pendant cinq à six mois, sans atteindre le stade prénymphe. Lorsque les ouvrières deS. fugax et les larves deL. nylanderi furent soumises ensemble à un hivernage préalable, elles donnèrent les mêmes résultats que sans hivernage. La présence d'une jeune reine deSolenopsis fut défavorable aux larves deLeptothorax.Inversement,L. nylanderi fut capable d'élever, à la température de 22°C, des larves deS. fugax et de les amener jusqu'au stade adulte. En présence de leurs propres larves, les ouvrières deL. nylanderi détruisirent tapidement toutes les larves deS. fugax introduites dans leur nid. D'autre part, un jeune couvain deLeptothorax remplaçait plus ou moins rapidement les larves deLeptothorax enlevées au préalable; sa présence était alors défavorable au développement des larves deSolenopsis. Un hivernage en début d'expérience fut plutôt favorable auxS. fugax, de même que la présence d'une reine féconde deLeptothorax. LesSolenopsis ainsi obtenus n'ont pas vécu plus de sept semaines. Ils étaient tous de caste ouvrière et de taille très petite.

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Summary When its own eggs and larvae missed,Solenopsis fugax bred larvae ofLeptothorax nylanderi, at a temperature of 22°C. TheSolenopsis workers killed some of this larvae and fed the others; these slowly grew bigger during five or six months but never reached the pre-pupa stage. The result was the same if the workers ofS. fugax and the larvae ofL. nylanderi overwintered together or not at all. A youngSolenopsis queen being there was noxious to the larvae ofLeptothorax.On the contrary,L. nylanderi has been able to breed larvae ofS. fugax up to the imago stage, at a temperature of 22°C. When its own larvae were in the nest, together with larvae ofS. fugax, the workers ofL. nylanderi killed the larvae ofS. fugax. On the other hand, new eggs and young larvae ofLeptothorax had to replace, more or less quickly, the larvae which had been taken away, and that was noxious to the growth ofSolenopsis larvae. An overwintering at the beginning of the experiment was rather favourable toS. fugax as was the presence of a fecundLeptothorax queen. TheSolenopsis thus obtained lived no longer than seven weeks. They all were workers and very small.

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S. Fugax L. Nylanderi 22° . Leptothorax , , , , . . S. Fugax Leptothorax.,L. Nylanderi 22° S. Fugax . L. Nylanderi ( )Leptothorax ; S. Fugax Solenopsis, Leptothorax. S. Fugax . .

     

Comparative study of the γ-butyrolactone and pantolactone contents in cells and musts during vinification by three Saccharomyces cerevisiae races

Summary Changes in the -butyrolactone and pantolactone contents in yeast cells and musts during fermentation and subsequent flor veil formation of Sherry wines were studied. Saccharomyces cerevisiae race cerevisiae, S. cerevisiae race bayanus and S. cerevisiae race capensis were used. During the alcoholic fermentation (first 31 days), -butyrolactone contents in musts and yeast cells were similar for the three yeast races tested. In this period, pantolactone was excreted to the must by bayanus and capensis races, and it was not detected in cerevisiae race cells. During flor veil formation (31 to 134 days), bayanus and capensis races yield higher -butyrolactone and pantolactone contents than cerevisiae race in the wines. In the final wines, pantolactone contents were always lower than those of -butyrolactone.     

Photosynthetic oxygen evolution: H/D isotope effects and the coupling between electron and proton transfer during the redox reactions at the oxidizing side of Photosystem II

The oxygen evolving complex (OEC) of Photosystem II (PS II) incorporates a Mn-cluster and probably a further redox cofactor, X. Four quanta of light drive the OEC through the increasingly oxidized states S0 S1 S2 S3 S4 to yield O2 during the transition S4 S0. It has been speculated that the oxidation of water might be kinetically facilitated by the abstraction of hydrogen. This implied that the respective electron acceptor is deprotonated upon oxidation. Whether YZ and X fulfill this expectation is under debate. We have previously inferred a 'chemical' deprotonation of X based on the kinetics of proton release (Haumann M, Drerenstedt W, Hundelt M and Junge W (1996) Biochim Biophys Acta 1273: 237–250. Here, we investigated the rates of electron transfer and proton release as function of the D2O/H2O ratio, the pH, and the temperature both in thylakoids and PS II core particles. The largest kinetic isotope effect on the rate of electron transfer (factor of 2.1–2.4) and the largest pH-dependence (factor of about 2 between pH 5 and 8) was found on S2 S3 where X is oxidized. During the other transitions both factors were much smaller ( 1.4). Electron transfer is probably kinetically steered by proton transfer only during S2 S3. These results corroborate the notion that X serves as a hydrogen acceptor for bound water during S4 S0. We propose a consistent scheme for the final reaction with water to yield dioxygen: two two-electron (hydrogen) transfers in series with a peroxide intermediate.     

Das organische Gerüstwerk der Kalkschale des Schwanen-Eies

Zusammenfassung Kegel und Säulen der Schwanen-Eischale hinterlassen am Querschliff nach Entkalkung mit EDTA organisches (mucoproteides) Material als ein zusammenhängendes Gerüst, das sich mit Thionin metachromatisch färbt; ohne Demineralisierung oder wenigstens Anätzung bleibt Thionin an Schliffen und Bruchkanten der Schale wirkungslos. Das Lichtmikroskop zeigt an Schliffen nichts von dem organischen Material, es wurde während des Kristallwachstums fein zerteilt in Gitterlücken des Schalencalcits eingeschlossen. Es findet sich am stärksten angehäuft an den äueren und inneren Oberflächen der Kristall-individuen. In den Kegeln ist das Gerüst radial ausgebildet als die Loculi der Keile, und konzentrisch geschichtet, entsprechend den Lagen der Globularinklusionen, um deren jede herum Verdichtung der organischen Substanz statthat. In den inneren Säulen folgt das organische Gerüst dem Rhombenmuster; die äueren Säulen sind arm an organischer Substanz, hier verbleibt nach der Entkalkung eine dünne laterale Oberflächenschicht.

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Summary The cones and columns of the swans egg shell leave behind after decalcification with EDTA an organic (mucoproteid) material in form of a continuous frame work stainable metachromatically with thionine. Without demineralisation or at least etching, thionine proves ineffectual in ground sections or breaking edges of the shell. In ground sections the light microscope demonstrates nothing of the organic material: it was inclosed during the crystal growth in submicroscopical lattice gaps of the calcite individuals. The organic material is chiefly accumulated in the outer and inner surfaces of the crystals. In the cones the organic frame work is developed radially as the loculi of the wedges and concentrically layered corresponding with the globular inclusions, concentrated in the circumference of each. In the inner columns the organic material follows to the rhomb pattern. The outer columns after decalcification only leave behind a thin lateral organic sheath.

     

Preparation of a series of sulfated tetrasaccharides from shark cartilage chondroitin sulfate D using testicular hyaluronidase and structure determination by 500 MHz1H NMR spectroscopy

Six tetrasaccharide fractions were isolated from shark cartilage chondroitin sulfate D by gel filtration chromatography followed by HPLC on an amine-bound silica column after exhaustive digestion with testicular hyaluronidase. Their structures were determined unambiguously by one- and two-dimensional 500 MHz¹H NMR spectroscopy in conjunction with HPLC analysis of chondroitinase AC-II digests of the tetrasaccharides. One fraction was found to contain two tetrasaccharide components. All the seven tetrasaccharides shared the common core structure GlcA1-3GalNAc1-4GlcA1-3GalNAc with various sulfation profiles. Four were disulfated comprising of two monosulfated disaccharide units GlcA1-3GalNAc(4-sulfate) and/or GlcA1-3GalNAc(6-sulfate), whereas the other three were hitherto unreported trisulfated tetrasaccharides containing a disulfated disaccharide unit GlcA(2-sulfate)1-3GalNAc(6-sulfate) and a monosulfated disaccharide unit GlcA1-3GalNAc(4-or 6-sulfate). These sulfated tetrasaccharides were demonstrated to serve as appropriate acceptor substrates for serum -N-acetylgalactosaminyltransferase, indicating their usefulness as authentic oligosaccharide substrates or probes for the glycobiology of sulfated glycosaminoglycans.Abbreviations NFU National formulary unit - COSY correlation spectroscopy - HOHAHA homonuclear Hartmann-Hahn - 1D or 2D one- or two-dimensional - IdoA l-iduronic acid - GlcA d-gluco-4-enepyranosyluronic acid - Di-0S GlcA1-3GalNAc - Di-4S GlcA1-3GalNAc(4-sulfate) - Di-4S GlcA1-3GalNAc(4-sulfate) - Di-6S GlcA1-3GalNAc(6-sulfate) - Di-6S GlcA1-3GalNAc(6-sulfate) - Di-diS _d GlcA(2-sulfate)1-3GalNAc(6-sulfate) - Di-diS_E GlcA1-3GalNAc(4, 6-disulfate) - U G, U, 2S, 4S, and 6S represent GlcA, GalNAc, GlcA, 2-O-sulfate, 4-O-sulfate, and 6-O-sulfate, respectively