NADH is specifically channeled through the mitochondrial porin channel in Saccharomyces cerevisiae

In many kinds of permeabilized cells, the restriction of metabolite diffusion by a mitochondrial porin closed state has been shown to control the respiration rate. However, since in isolated mitochondria the porin appears to be always open, the physiological relevance of a putative regulation via this channel status is now a subject of discussion. In Saccharomyces cerevisiae, in which some of the NADH dehydrogenase active sites are facing the intermembrane space, this regulatory mechanism might play an important role for the regulation of the cytosolic redox status. Using permeabilized spheroplasts from wild-type and porin-deficient mutant, we show that the NADH produced in the cytosol is channeled to the mitochondrial NADH dehydrogenases through a metabolic network involving the porin channel. Thus, the control exerted by the porin (i.e., open or closed state) seems to be determined through its participation or not in organized metabolic networks.     

Inactivation of voltage-dependent calcium current in an insulinoma cell line

We have studied the mechanism of Ca current inactivation in the -cell line HIT-T15 by conventional and perforated patch recording techniques, using two pulse voltage protocols and a combination of current and tail current measurements. In 5 mM Ca, from a holding potential of - 80 mV, the maximum current showed a complex time course of inactivation: a relatively fast, double exponential inactivation (_h1 12 ms and _h2 60 ms) and a very slowly inactivating component ( > 1 s). The faster component (_h1) was due to the voltage-dependent inactivation of a low-threshold-activated (LVA), T-type current, which deactivates more slowly ( 3–5 ms) than the other components ( 0.2–0.3 ms). The intermediate component (_h2) was due to the Ca-dependent inactivation of a portion of the high-threshold-activated (HVA) current. A saturating dose of the dihydropyridine (DHP) nifedipine (10 M) did not affect the LVA current, but inhibited by 68 ± 5% the transient, Ca-sensitive portion of the HVA current and by 33 ± 12% the long lasting component. We suggest that three components of the calcium current can be resolved in HIT cells and the main target of DHPs is a HVA current, which inactivates faster than the DHP-resistant HVA component and does so primarily through calcium influx. Correspondence to: C. Marchetti     

Elektronenmikroskopische Untersuchungen über Bildung und Struktur der Zygotenwand beiMicrasterias papillifera (Desmidiaceae)

Zusammenfassung Die Feinstruktur von Mesospor und Endospor reifer Zygoten vonMicrasterias papillifera wurde untersucht. Das für die Resistenz der Zygoten gegenüber ungünstigen Umweltbedingungen wichtige Mesospor besteht aus vier Schichten von unterschiedlicher Elektronendichte. Es ist insgesamt 460–500 nm dick. Die Schichten Mes a und Mes c bestehen aus akkrustierten, dicht gepackten globulären Elementen eines Stoffes, der dem Sporopollenin ähnlich ist. Die Schicht Mes b zeigt ein fibrilläres Grundgerüst, wahrscheinlich aus Zellulose, in das Stoffe inkrustiert sind, die nicht mit denen der Schichten Mes a und Mes c identisch, aber gegen Abbauversuche ähnlich résistent sind.Die Schicht Mes d ist eine Übergangsschicht zum Endospor. Zwischen die Zellulose-Mikrofibrillen in Streutextur sind isolierte Partikel des Materials der Schicht Mes c eingestreut. Das etwa 650 nm dicke Endospor ist eine Zelluloseschicht mit Streutextur. Es wird als Wand der sogenannten Keimblase bei der Zygotenkeimung nach Sprengung von Exospor und Mesospor stark gedehnt.

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Electron microscopical investigations on the structure and formation of the zygote wall inMicrasterias papillifera (Desmidiaceae) II. The structure of the mesospore and the endospore

Summary The mesospore (460–500 nm thick) which is responsible for the resistance of the zygote against desiccation during its resting period, consists of four layers of different electron density. The layers Mes a and Mes c are composed of densely packed amorphous globular elements of a substance resembling sporopollenine. The layer Mes b has a fibrillar, probably cellulosic frame. It is incrusted by a substance which is not identical with that of Mes a or Mes c but which shows a comparable resistance against degradation.The layer Mes d contains isolated particles such as in Mes c and cellulose microfibrils of the endospore. The endospore (650 nm thick) has foliate texture. This layer surrounds the protoplast after it has escaped from the ruptured exospore and mesospore during zygospore germination.

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Der Deutschen Forchungsgemeinschaft dank ich f:ur Sachbeihilfe.     

Bilan nutritionnel au cours du développement de l'ectoparasite grégaire Dinarmus vagabundus et du solitaire Dinarmus basalis

Résumé Nos méthodes expérimentales permettent l'isolement d'une larve de sexe déterminé par hôte de l'ectoparasite grégaire Dinarmus vagabundus et du solaitire, D. basalis. Des hôtes porteurs de 3 à 8 larves par hôte de D. vagabundus sont aussi isolés. Dans ces conditions la quantité de nourriture disponible est la même pour toutes les densités larvaires étudiées.Les larves élevées en solitaire des deux espèces assimilent une quantité de nourriture significativement supérieure à celle assimilée par les . Ceci conduit à des adultes de poids moyen supérieur à celui des . Le poids moyen des et des de D. vagabundus diminue significativement aux fortes densités larvaires. L'intensité de la liaison entre la quantité de nourriture assimilée et la biomasse produite s'affaiblit au fur et à mesure que la densité larvaire par hôte augmente.Les de D. vagabundus de poids moyen (0,42 mg) engendrent deux fois et demi plus de descendants que les lilliputiennes (0,20 mg) émergées d'hôtes à forte densité larvaire. Celles de D. basalis (0,65 g) sont moins prolifiques que les de D. vagabundus.     

Changes of [3H]Muscimol Binding and GABAA Receptor β2-Subunit mRNA Level by Tolerance to and Withdrawal from Pentobarbital in Rats

Effects of continuous pentobarbital administration on binding characteristics of [³H]muscimol were examined by autoradiography, and levels of GABA_A receptor 2-subunit mRNA were investigated by in situ hybridization histochemistry in the rat brain. In order to eliminate the induction of hepatic metabolism by systemic administration of pentobarbital, an i.c.v. infusion model of tolerance to and withdrawal from pentobarbital was used. An experimental model of barbiturate tolerance and withdrawal was developed using i.c.v. infusion of pentobarbital (300 g/10 l/hr for 7 days) by osmotic minipumps and abrupt withdrawal from pentobarbital. The levels of [³H]muscimol binding were elevated in cingulate of frontal cortex (46%) and granule layer of cerebellum (32%) of rats 24-hr after withdrawal from pentobarbital, while it was only elevated in cingulate (58%) of tolerant rats. The GABA_A receptor 2-subunit mRNA was increased in the withdrawal rats only: in the cortex (9–14%), hippocampus (15–21%), inferior colliculus (21%), and granule layer of cerebellum (24%). These results show the involvement of GABA_A receptor and its 2-subunit up-regulations in pentobarbital withdrawal rats, and suggest that the levels of [³H]muscimol binding and GABA_A receptor 2-subunit mRNA are altered in a region-specific manner during pentobarbital withdrawal.     

Microbial metabolism of alkylbenzene sulphonates the oxidation of key aromatic compounds by a Bacillus

A Bacillus species originally elected for growth at the expense of alkylbenzene sulphonate detergents was found to metabolise a wide range of aromatic compounds. p-Hydroxybenzoate (PHB) was initially hydroxylated to protocatechuate (PCA) i.e. 3,4-dihydroxybenzoate, which was oxidatively cleaved to succinate and acetyl-CoA by a classical ortho cleavage pathway initiated by a substrate-specific 3, 4-oxygenase: no evidence of an alternative meta cleavage pathway was detected. Several key enzymes of this ortho cleavage pathway were induced by growth of the Bacillus on either PHB or PCA. Both PHB and PCA were able to act as sole source of carbon for energy and overall growth of the microorganism.In strict contrast, the higher homologue p-hydroxyphenylacetate (PHPA), after initial hydroxylation to 3, 4-dihydroxyphenylacetate (DHPA), was oxidatively cleaved to 4-carboxymethyl-2-hydroxymuconic semialdehyde (CMHMS) by a meta cleavage catalysed by a substrate-specific 2, 3-oxygenase: no evidence of an alternative ortho cleavage was detected. Several lines of evidence suggested that CMHMS was not further metabolised by the Bacillus and accumulated in the growth medium. Both PHPA and DHPA were unable to act as sole source of carbon for energy and overall growth.The implication of the occurrence in a single bacterium of two separate oxidative pathways catalysing the cleavage of different aromatic nuclei have been discussed.     

Intraorbital cerebrospinal fluid outflow and the posterior uveal compartment of the hamster eye

Summary An ultrastructural and tracer study was undertaken to determine normal outflow pathways of cerebrospinal fluid (CSF) at the terminal subarachnoid space (SAS) of the optic nerve. In the morphological studies, the optic nerve dura and arachnoid were found to be continuous with the sclera of the eye beyond the optic nerve SAS. The pia mater is continuous with the inner sciera and the lamina fusca of the eye. Montages and serial sections demonstrated that the distal SAS is divided into numerous tortuous channels to form an arachnoidal trabecular meshwork. Spaces of this meshwork continue into microcanals which bypass the outer arachnoid barrier layers of the optic nerve meninges to reach the sclera and posterior intraorbital connective tissue. Ferritin infused into the cisterna magna entered the optic nerve SAS within 1 min and reached arachnoidal trabecular meshwork channels and the microcanals within 8 min. It then passed into intraorbital connective tissue spaces at the posterior pole of the eye. Ferritin appeared to be blocked by the lamina fusca and a newly discovered posterior compact zone which together prevented its entrance into the choroidal interstitium. These observations suggest that a subarachnoidal-scleral-orbital outflow pathway provides a route for CSF drainage from the optic nerve SAS to intraorbital connective tissue. The previously described posterior uveal compartment in the hamster eye (Kelly et al. 1983) appears to be relatively isolated from this subarachnoidal-scleral-orbital CSF outflow.Parts of this work have been presented at the 1984 meetings of the American Association of Anatomists (Shen 1984).     

Intracellular sites of the synthesis of sweet potato mitochondrial F1ATPase subunits

Summary Five subunits (-, -, -, - and -subunits) of the six -and -subunits) in the F₁ portion (F₁ATPase) of sweet potato (Ipomoea batatas) mitochondrial adenosine triphosphatase were isolated by an electrophoretic method. The - and -subunits were not distinguishable immunologically but showed completely different tryptic peptide maps, indicating that they were different molecular species. In vitro protein synthesis with isolated sweet potato root mitochondria produced only the -subunit when analyzed with anti-sweet potato F₁ATPase antibody reacting with all the subunits except the -subunit. Sweet potato root poly(A)⁺RNA directed the synthesis of six polypeptides which were immunoprecipitated by the antibody: two of them immunologically related to the -subunit and the others to the - and -subunits. We conclude that the -subunit of the F₁ATPase is synthesized only in the mitochondria and the -, - and -subunits are in the cytoplasm.     

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.     

Binding of the galactose-specificPseudomonas aeruginosa lectin,PA-I,to glycosphingolipids and other glycoconjugates

The carbohydrate-binding specificity ofPseudomonas aeruginosa lectin I (PA-I) in iodinated or biotinylated form was studied. A large number of glycosphingolipids, as well as some glycoproteins and neoglycoproteins were used as ligands. Also, inhibition by free saccharides of PA-I binding to glycosphingolipids was tested. It was found that the lectin binds most strongly to terminal and nonsubstituted Gal3Gal- or Gal4Gal-structures.Abbreviations PA-I Pseudomonas aeruginosa lectin I - Cer ceramide - lactosylceramide Gal4GlcCer - iso globotriaosylcerami Gal3Gal4GlcCer - globotriaosylceramide Gal4Gal4GlcCer - globoside or globotetraosylceramide GalNAc3Gal4Gal4GlcCer - Forssman glycolipid GalNAc3GalNAc3Gal4Gal4GlcCer - P1 glycolipid Gal4Gal4GlcNAc3Gal4GlcCer - lactoneotetraosylceramide Gal4GlcNAc3Gal4GlcCer - B5 glycolipid Gal3Gal4GlcNAc3Gal4GlcCer - gangliotetraosylceramide Gal3GalNAc4Gal4GlcCer - GM1 Gal3GalNAc4(NeuAc3)Gal4GlcCer - RBC red blood cells - BSA bovine serum albumin - PBS phosphate-buffered saline - SDS sodium dodecyl sulfate - TLC thin-layer chromatography - HPLC high pressure liquid chromatography - MS mass spectrometry - FAB fast-atom bombardment - EI electron impact     

Aflatoxin influences on seed germination and root elongation by two cultivars of Glycine max and uptake of 65 Zn-ZnCl2 by the cultivars

Maximum inhibition of Glycine max, cv. Essex seed germination occurred at 10 g/ml following 72 hr imbibition in constant light. Seeds imbided 108 hr in constant darkness at this concentration showed a 20% rise in germination over that of the control. Imbibition of G. max, cv. Williams seeds in either light or dark for 96 hr did not suppress germination. Imbibition of Essex seeds in either light or dark at 2.5 through 10 g/ml stimulated root elongation except for 10 g/ml at 96 hr (light). Maximum inhibition of Williams root elongation under constant light was at 48 and 72 hr with 10 g/ml. Statistically significant differences in cotyledon, leaf and stem lengths between non-treated (NT) and treated (T) seedlings were not found except for Williams stem length at 2.5 / ml. Root elongation was stimulated 1.2- and 1.1-folds, respectively, at 5.0 (Essex) and 2.5 (Williams) g/ml. Toxin at 2.5 through 10.0 g/ml did not markedly alter either cotyledon or leaf widths with the exception of Williams leaf width at 2.5 g/ml. Medium supplementation with 2.5 through 10.0 g/ml resulted in cotyledon, leaf and root weight enhancements for Essex seedlings. Stem weight was not markedly affected. An 18% rise in Williams cotyledon weight above that of the control was seen at 2.5 g/ml. Williams leaf weights were increased 1.75- and 1.25-folds, respectively, at 2.5 and 10.0 g/ml. Aflatoxin B₁, at 2.5 g/ml promoted Williams stem and root elongation 1.20- and 1.09-folds, respectively. Most of the radioactivity from ⁶⁵Zn-ZnCl₂ recovered within organs was found within Essex roots for both T and NT seedlings. A higher amount of radioactivity was recovered within roots at each toxin concentration than was without toxin. However, this was not statistically significant. Significant differences in the distribution of radioactivity within roots between NT and T Williams seedlings were not observed. Generally, AFB₁ failed to affect significantly these two varieties of soybeans based on the tests relating to germination, growth and radiolabel uptake.     

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     

Effect of androgens on histochemical fibre type

Summary The temporal muscles of the guinea pig show a sexual differentiation reflected in their histochemical enzyme pattern. Using histochemical methods for mitochondrial (SDH, -GPDH), and glycolytic enzymes (phosphorylase, LDH) it could be shown, that in adult animals the male muscle is a white muscle with marked activity of glycolytic enzymes, the female muscle a red muscle displaying high activity of mitochondrial enzymes. This differential enzyme pattern can be converted by the application of testosterone to the female type during the postnatal development. The male sex hormone thus affects the histochemical enzyme pattern of the muscle, converting the red, female into a white, male muscle in the female guinea pig.     

Expression of a chimeric stilbene synthase gene in transgenic wheat lines

A chimeric stilbene synthase (sts)gene was transferred into wheat. Stilbene synthases play a role in the defence against fungal diseases in some plant species (e.g. groundnut or grapevine) by producing stilbenetype phytoalexins like resveratrol. Resveratrol is also claimed to have positive effects to human health. Embryogenic scutellar calli derived from immature embryos of the two commercial German spring wheat cultivars Combi and Hanno were used as target tissue for cotransformation by microprojectile delivery. The selectable marker/reporter gene constructs contained the bargene either driven by the ubiquitinpromoter from maize (pAHC 25, also containing the uidAgene driven by the ubiquitinpromoter), or by the actinpromoter (pDM 302) from rice. The cotransferred plasmid pStil 2 consisted of a grapevine stscoding region driven by the ubiquitin promoter. Eight transgenic Combi and one Hanno T_Oplant were obtained and, except one Combi T_Oplant, found to be cotransformants due to the integration of both the stsgene and the selectable marker or reporter genes. Expression of the stsgene was proven by RTPCR, and, for the first time, by detection of the stilbene synthase product resveratrol by HPLC and mass spectrometry. The stsgene was expressed in four of the seven transgenic Combi T_{_o}plants. Two of the respective T₁progenies segregated in a Mendelian manner were still expressing the gene. Investigations into methylation of the stsgene showed that in three nonexpressing progenies inactivation was paralleled by methylation.     

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.     

Factors determining the abundance and distribution of rodents in a shrub-steppe ecosystem: the role of shrubs

Summary This study addressed the relative importances of shrub resources on a rodent community in a sagebrush dominated shrub-steppe ecosystem in southwestern Wyoming. Direct effects of shrubs (i.e., providing rodents with food and cover) were assessed by removing shrubs from a 1.25 ha study plot and monitoring both rodent populations and their food resources. Shrub architecture and shrub-related food resources were found to be unimportant to deermice (Peromyscus maniculatus), Great Basin pocket mice (Perognathus parvus) northern grasshopper mice (Onychomys leucogaster) and Uinta ground squirrels (Spermophilus armatus), as shrub removal caused no significant changes in population sizes, sex ratios or age structure. Least chipmunks (Eutamias minimus) responded to shrub removal by leaving the plot and moving into adjacent shrubland. The montane vole (Microtus montanus) population showed a slight increase following shrub removal. Shrub removal did not alter the abundance of major rodent food resources on the plot (percent cover of herbaceous vegetation, soil seed reserves and ground-dwelling arthropods). Micrometeorological data suggested that shrubs did not significantly ameliorate a nocturnal rodent's micro-climate, but may have affected diurnal rodents' thermal loading rates by removing shade. While shrub architecture and food resources do not directly affect most of the rodents in this shrub-steppe ecosystem, shrubs may be important to rodents in a long-term time frame. Shrubs provide safe sites for germination and growth of herbaceous vegetation, thereby enhancing the diversity of the potential rodent food resources.     

The respiratory system of the femaleVarroa jacobsoni (Oudemans): its adaptations to a range of environmental conditions

The morphology of the respiratory system of the femaleVarroa jacobsoni (Oudemans, 1904) is described. The mobile, appendage-like, emergent peritreme may be raised to lie against the ventral integument or lowered between the third and fourth pair of legs. It is raised when the mite is submerged in the liquid food of the host's brood chamber, where respiration occurs via an external plastron, formed by an airfilm trapped between the rough cuticle of the ventral integument and the retracted legs. The peritreme is also raised when the mite is outside the hive in sub-saturated air, to reduce water vapour transpiration, and it is lowered in the carbon-dioxide-rich and water-saturated hive atmosphere, where it facilitates rapid removal of carbon dioxide. Thus gaseous exchange in the female mites may be adjusted by the position of the peritreme.Key to captions a ascending limb of peritrematic groove - c hollow (haemocoel) core of peritreme - d descending limb of peritrematic groove - e endocuticle - f flange of the inner stigmatic orifice - g peritrematic groove - i funnel of inner stigmatic orifice - k circum-spiracular pocket at base of peritreme - 1 peritrematic slit - m micropapillae - n fringe of marginal setae - o outer stigmatic orifice (=stigma) - p emergent peritreme - r rough integument at bases of leg coxae - s stigmatic atrium - t tracheal atrium - u outer lip of outr stigmatic orifice - v inner lip of outer stigmatic orifice - x tracheal trunks - z thick cuticle of peritrematic groove - 1,2,3,4 numbers of legs or leg coxae     

Spermatogonia and the cycle of the seminiferous epithelium in the nine-banded armadillo

Summary The cycle of the seminiferous epithelium of the nine-banded armadillo can be divided into ten stages. As in most mammals, only one stage is observed per tubular cross-section. The process of spermiogenesis can be divided into thirteen steps according to the development of the acrosomal system and the flagellum. Four generations of spermatogonia are observed in the germinal epithelium: 1) stem cells, 2) type A, 3) intermediate, and 4) type B spermatogonia. The stem cell is characterized by a highly irregular nucleus and the presence of glycogen in its cytoplasm. The type A spermatogonium contains an oblong nucleus with one or two shallow infoldings of the nuclear membrane. The intermediate spermatogonium contains an ovoid nucleus characterized by one or two nuclei and heterochromatin scattered in the nucleoplasm. The nucleus of the type B spermatogonium is more spherically shaped with a centrally placed nucleolus and heterochromatin associated with the nuclear envelope.The author wishes to acknowledge the technical assistance of Teri Lane     

Backbone dynamics of the cytotoxic ribonuclease alpha-sarcin by 15N NMR relaxation methods

The cytotoxic ribonuclease -sarcin is a 150-residue protein that inactivates ribosomes by selectively cleaving a single phosphodiester bond in a strictly conserved rRNA loop. In order to gain insights on the molecular basis of its highly specific activity, we have previously determined its solution structure and studied its electrostatics properties. Here, we complement those studies by analysing the backbone dynamics of -sarcin through measurement of longitudinal relaxation rates R₁, off resonance rotating frame relaxation rates R₁, and the ¹⁵N¹HNOE of the backbone amide ¹⁵N nuclei at two different magnetic field strengths (11.7 and 17.6 T). The two sets of relaxation parameters have been analysed in terms of the reduced spectral density mapping formalism, as well as by the model-free approach. -Sarcin behaves as an axial symmetric rotor of the prolate type (D/D=1.16 ± 0.02) which tumbles with a correlation time _m of 7.54 ± 0.02 ns. The rotational diffusion properties have been also independently evaluated by hydrodynamic calculations and are in good agreement with the experimental results. The analysis of the internal dynamics reveals that -sarcin is composed of a rigid hydrophobic core and some exposed segments which undergo fast (ps to ns) internal motions. Slower motions in the s to ms time scale are less abundant and in some cases can be assigned to specific motional processes. All dynamic data are discussed in relation to the role of some particular residues of -sarcin in the process of recognition of its ribosomal target.     

The role of muscle development in the transition to endothermy in nestling bank swallows,Riparia riparia

Summary We examined the transition from ectothermy to endothermy in nestling bank swallows (Riparia riparia) by measuring the peak metabolic response to cold (PMR) in groups of nestlings. Additionally aerobic capacity, as assessed by citrate synthase activity (CS), and contractile function, as assessed by myofibrillar ATPase activity (mATPase) were measured in the pectoralis and mixed leg muscles during development. During the first 65% of their growth (from 2–12 g) bank swallows do not increase their metabolic rate in response to cold (Fig. 1). Between 12 and 16 g the PMR increased from 4 to more than 10 ml O₂ (g·h)^–1. Citrate synthase activity increased throughout development, starting at 20 moles (min·g fresh mass)^–1 in both tissues and increasing to 150 and 50 moles (min·g)^–1 in the pectoralis and leg muscles, respectively (Fig. 5). The augmented aerobic capacity combined with large increases in muscle mass undoubtedly contributes to the improved thermoregulatory abilities of older nestlings. However, muscle mass and aerobic capacity increase continuously and do not show the sharp transition noted in PMR. In the leg muscle mATPase activity is constant throughout growth, but in the pectoralis muscle it undergoes an abrupt increase from 0.5 moles (min·mg myofibrillar protein)^–1 in animals weighing less than 12 g to 0.9 moles (min·mg)^–1 in nestlings weighing more than 15 g (Fig. 6). The similar pattern of development of PMR and mATPase suggests a critical role for muscle development in the transition to endothermy in this species.Abbreviations CS citrate, synthase - mATPase myofibrillar adenosine triphosphatase - PMR peak metabolic rate during cold stress - rate of oxygen consumption