Spatial distribution of brood-bearing females of limnetic species of Cladocera

In this study, the spatial distribution of brood-bearing females of five species of limnetic cladocerans (Daphnia cucullata, D. longispina, Bosmina coregoni, B. longirostris, Diaphanosoma brachyurum) in the deep mesotrophic lake in relation to the predation pressure of planktivorous fish (roach Rutilus rutilus, perch Perca fluviatilis, catfish Ictalurus nebulosus, white fish Coregonus albula, bleak Alburnus alburnus), and planktonic invertebrates (cyclopoids Mesocyclops leuckartii, Thermocyclops oithonoides, T. crassus, and cladoceran Leptodora kindtii) as well as some environmental variables was estimated. Most cladocerans showed apparent differences in horizontal distribution (ANOVA F = 0.2–0.45, P < 0.05) in the littoral zone and lack of such differences in the pelagic zone (F = 0.07–0.13, P > 0.05). Vertical distribution of most species, in turn, showed a clear pattern in the pelagic zone (F = 0.31–0.39, P < 0.05) and less regularities in the littoral zone (F = 0.15–029, P > 0.05). The differences in spatial distribution of non-predated and predated species suggest that predation pressure, but not predatory type, was an important factor structuring their distribution. Other factors that affected their distribution were conductivity, dissolved oxygen, TOC and macrophyte biomass; however, most of those variables better explained the distribution of brood-bearing cladocerans in the vertical than horizontal aspect.     

Studies of the morphology of chromosomes of some selected species

Karyotypes of twelve species from twenty-four localities in southern Moravia and one locality in Slovakia were investigated. Their counts or karyotypic formulae are as follows:Chenopodium foliosum (Moench) Ascherson: K (2n)=18=16 A^m+2 B^sm;Astragalus austriacus Jacq.: K (2n)=16=8 A^m+8 B^sm;Astragalus exscapus L.: K (2n)=16=10 A^m+4 B^sm+2 C^st;Astragalus cicer L.: K (2n)=64;Astragalus onobrychis L.: K (2n=64 and K (2n)=64+1;Vicia dumetorum L.: K (2n=14=10 A^m+4 B^sm;Vicia sylvatica L.: K (2n)=14=2 A^m+10 B^sm+2 C^st;Vicia pisiformis L.: K (2n)=12=8 A^m+4 B^sm;Vicia cassubica L.: K (2n)=12=4 A^m+6 B^sm+2 C^st;Vicia cracca L. (from five localities in southern Moravia): K (2n)=28=4 A^m+12 B^sm+12 C^st and K (2n)=28+1=5 A^m+12 B^sm+12 C^st;Vicia cracca L. (from one locality in Slovakia): K (2n)=14=2 A^m+6 B^sm+6 C^st;Vicia tenuifolia Roth: K (2n)=24=4 A^m+16 B^sm+4 C^st;Serratula lycopifolia (Vill.) Kern.: K (2n)=60.     

Synthesis of Fragments of the Peptide Component of Pseudobactin

Pseudobactin is a structurally complex and physiologically important siderophore (microbial iron chelator) from Pseudomonas putida- fluorescens. Various fragments of the unusual peptide component of pseudobactin listed below were prepared by solution-phase peptide synthesis. L -Lys· D -threo-β-OH Asp· L -Ala· D -allo-Thr· L -Ala L -Lys· D -threo-βOH Asp· L -Ala· D -allo-Thr D -threo-β-OH Asp· L -Ala· D - allo-Thr· L -Ala· D -N-OH-cycloOrn D -threo-β-OH-Asp· L -Ala· D -allo-Thr· L -Ala L -Ala· D -allo-Thr· L - Ala· D -N-OH-cycloOrn A class of related peptides named pseudomycins have shown promising antifungal activity. To examine if these peptide fragments above would elicit similar activity, the fragments were tested and found to have no antifungal activity in limited bioassays.     

Comparison of the profiles of non-glycosylated triterpenoids from leaves of plants of selected species of genus Dioscorea

Remarkable qualitative and quantitative differences in non-glycosylated triterpenoid profiles of twelve Dioscorea spp. leaves were demonstrated with the use of GC–MS/FID analysis. The total content of tetracyclic triterpenoids and their esters ranged from 397 μg/g of dry leaf weight in D. bulbifera to 762 μg/g d.w. in D. discolor and 777 μg/g d.w. in D. alata. Three main phytosterols, i.e. campesterol (1), sitosterol (2) and stigmasterol (3) were found in extracts from all analyzed species in total amount ranging from 316 μg/g in D. bulbifera to 676 μg/g of dry leaf weight in D. hispida, with either sitosterol (2) or stigmasterol (3) as predominant in the profile. Extracts from D. hispida and D. purpurea leaves were distinguished from the others by particularly high amount of campesterol (1). In the majority of the species, except for D. caucasica, other tetracyclic triterpenoids were found: cycloartanol (4), 24-methylenecycloartanol (5) and cycloeucalenol (6). Less common steroids, stigmastan-3-en-6β-ol (7) and ergosta-7,22-dien-3β-ol (8) were detected in D. japonica. The significant content (992 μg/g) of pentacyclic triterpenoids of ursane, oleanane, taraxastane and taraxerene (friedooleanane)-type carbon skeletons, i.e. α-amyrin (9), β-amyrin (10), taraxasterol (11) and taraxerol (12), respectively, was found in D. caucasica. The obtained results supplement the knowledge of biochemical diversity of Dioscorea genus.     

Analysis of applicability of triplet-state emission of molecular hydrogen for spectral diagnostics of a DC discharge

The applicability of emission of the N ³Λ_σ triplet states of molecular hydrogen for spectral diagnostics of the positive column of a dc glow discharge in hydrogen at translational gas temperatures of 360–600 K, specific absorbed powers of 0.8–4.25 W/cm, gas pressures of p = 0.3–15.0 Torr, reduced fields of E/N = 30–130 Td, and electron densities of n _e = 4.0 × 10⁹–6.5 × 10¹⁰ cm^–3 is analyzed by using an advanced level-based semi-empirical collisional?radiative model. It is found that secondary processes make the main contribution to the population and decay of the N ³Λ_σ = a ³Σ⁺ _g , c ³Π _u , g ³Σ⁺ _g , h ³Σ⁺ _g , and i ³Π _g triplet states. The dipole-allowed transitions e ³Σ⁺ _g → a ³Σ⁺ _g , f ³Σ⁺ _g → a ³Σ⁺ _g , g ³Σ⁺ _g and k ³Π _u → a ³Σ⁺ _g can be used for spectral diagnostics of a dc discharge within a simplified coronal model.     

Effects of different batches of iodine on properties of I-hFSH and characteristics of radioligand-receptor assays

Radioiodination of highly purified human follicle-stimulating hormone (hFSH) (4000 IU/mg) was performed every other week for 23 weeks using 2 mCi carrier free Na ¹²⁵I (Amersham Corp., 15 mCi/μg I₂) in the presence of lactoperoxidase. Incorporation of ¹²⁵I into hFSH was determined by the method of [7.]Biochem. J. 89, 114). Hormone binding was studied in vitro under steady-state conditions (16 h, 20°C) using different calf testis membrane preparations having similar receptor characteristics. Each ¹²⁵I-hFSH preparation was characterized for maximum bindability, specific activity of bindable radioligand as determined by self-displacement analysis, and by determination of K_a and R_t. Incorporation of ¹²⁵I into FSH was relatively constant over the large number of experiments (62.4 ± 6.4 μCi/μg; n = 23). By comparison, however, specific radioactivity of the receptor bindable fraction of ¹²⁵I-hFSH was related to the lot of ¹²⁵I utilized, and was significantly (P ≤ 0.01) lower and more variable (28.7 ± 10.5 μCi/μg). Maximum bindability of ¹²⁵I-hFSH was not correlated to specific activity (r = 0.06) but was negatively correlated to hFSH ¹²⁵I incorporation (r = −0.47; P ≤ 0.05). These observations demonstrate the need to assess the quality of each batch of radioligand before undertaking radioligand-receptor assays and suggest that differences in Na¹²⁵I lots affect specific radioactivity of the radioligand and its receptor binding characteristics.     

Peculiarities of Selectivity of Three Subtypes of Low-Threshold T-Type Calcium Channels

Despite the progress in studies of the properties and functions of low-threshold calcium channels (LTCCs) [1], the mechanisms of their selectivity and permeability remain unstudied in detail. We performed a comparative analysis of the selectivity of three cloned pore-forming LTCC subunits (α1G, α1H, and α1I) functionally expressed in Xenopus oocytes with respect to bivalent alkaline-earth metal cations (Ba²⁺, Ca²⁺, and Sr²⁺. The relative conductivities (G) of these channels were determined according to the amplitudes of macroscopic currents (I) and potentials of zero currents (E). The currents were recorded after preliminary intracellular injection of a fast calcium buffer, BAPTA, in order to suppress the endogenous calcium-dependent chloride conductivity. Channels formed by α1G subunits demonstrated the following ratios of the amplitudes of macroscopic currents and potentials of zero current: I _Ca:I _Ba:I _Sr = 1.00:0.75:1.12 and E _Ca ≈ E _Ba ≈ E _Sr. For channels that were formed by α1H and α1I subunits, these ratios were as follows: I _Ca:I _Ba:I _Sr = 1.00:1.20:1.17, E _Ca ≈ E _Ba ≈ E _Sr and I _Ca:I _Ba:I _Sr = 1.00:1.48: 1.45, E _Ca ≈ E _Ba ≈ E _Sr respectively. The different macroscopic conductivities and similar potentials of zero current typical of α1G and α1I channels indicate that, probably, various bivalent cations can in a differential manner influence the stochastic parameters of functioning of these channels. At the same time, channels formed by α1H subunits are characterized by more positive potentials of zero current for Ca²⁺. It seems possible that the selectivity of the above channels is determined by mechanisms that mediate the selectivity of most high-threshold calcium channels (more affine binding of Ca²⁺ inside the pore). Neirofiziologiya/Neurophysiology, Vol. 37, No. 4, pp. 319–329, July–August, 2005.     

Effect of tryptophan and of glucose on exercise capacity of horses

We hypothesized that central fatigue may have a role in limitingthe endurance capacity of horses. Therefore, we tested the effect ofinfusing tryptophan and/or glucose on endurance time and plasmaconcentrations of free tryptophan and other substrates thought toaffect tryptophan uptake into the brain of seven mares (3-4 yr ofage, 353-435 kg) that ran on a treadmill at 50% of maximalO₂ consumption to fatigue. Withuse of a counterbalanced crossover design, the horses were infused withtryptophan (100 mg/kg in saline solution) or a similar volume of salinesolution (placebo) before exercise. During exercise, horses receivedinfusions of glucose (2 g/min, 50% wt/vol) or a similar volume ofsaline. Thus the treatments were 1)tryptophan and glucose (T & G), 2) tryptophan and placebo (T & P), 3)placebo and glucose (P & G), and 4)placebo and placebo (P & P). Mean heart rate, hematocrit, andconcentration of plasma total solids before and during exercise weresimilar for all trials. Mean time to exhaustion was reduced (P < 0.05) for T & P and T & Gcompared with P & P [86.1 ± 6.9 and 87.1 ± 6.8 vs. 102.3 ± 10.3 (SE) min], whereas endurance for P & G(122.4 ± 11.9 min) was greater than for all other trials (P < 0.05). Compared withnontryptophan trials, during the tryptophan trials plasma prolactinincreased (P < 0.05) nearlythreefold before exercise and almost twofold early in exercise. Muscleglycogen concentrations were reduced(P < 0.05) below preexercise values in the P & G and P & P trials only. However, glucose infusions (P & G)did not affect (P > 0.05)concentrations of plasma free fatty acids or ratios of branched-chainamino acids to free tryptophan. In conclusion, tryptophan infusionreduced endurance time, which was consistent with the central fatiguehypothesis. The failure of glucose infusion to alleviate the effects oftryptophan and the absence of significant muscle glycogen reduction inthe tryptophan trials suggest that the early onset of fatigue in thetryptophan trials is not due to a lack of readily available substrate.

     

Influence of method of presentation on results of plant-host preference tests with two species of grasshopper

Leaf material of two Piper spp. (Piperaceae) was presented to two different generalist herbivores (Orthoptera: Acrididae) to determine whether method of leaf presentation in preference trials affects choice. For the grasshopper Leptomerinthoprora brevipennis, the method of presentation had no effect on preference ranking: more of Piper sancti-felicis was eaten than of P. arieianum, regardless of whether leaf material was presented as leaf disks, whole leaves supplemented with water, or whole leaves without water. In contrast, preference of the grasshopper Abracis flavolineata depended on presentation method. In that case, whole leaves of P. scancti-felicis supplemented with water were preferred over the same of P. arieianum, while dry weight consumption of leaf disks of P. arieianum was greater than that for disks of P. sancti-felicis. When presented with leaves not supplemented with water, A. flavolineata showed no significant preference for one plant species over another.

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Résumé Des feuilles provenant de deux espèces de Piper (Piperacae) ont été proposées à deux Orthoptères, Acrididae, herbivores généralistes, pour déterminer si la méthode de présentation des feuilles modifiait le choix. Pour Leptomerinthoprora brevipennis, la méthode de présentation est sans influence sur l'ordre de préférence; la consommation de Piper sancti-felicis est toujours supérieure à celle de P. arieianum, quelque soit le mode de présentation: disques de feuilles, feuilles entières humidifiées, ou feuilles entières sans addition d'eau. A l'opposé, l'ordre de préférence de Abracis flavolineata dépend du type de présentation; les feuilles entières humidifiées de P. sancti-felicis sont préférées à celles de P. arieianum proposées de la même façon, tandis que la consommation de disques secs de P. arieianum est plus importante que celle des disques secs de P. sancti-felicis. En présence de feuilles entières humidifiées, A. flavolineata ne manifeste aucune préférence.

     

Critical survey of the present stand of the production of perfect stage of organs of fructification in dermatophytes

Summary Very little progress had been made on the question whether organs of higher fructification occur in dermatophytes. Only two significant studies can be noted. One was the elaboration of the hypothesis byMatruchot &Dassonville that dermatophytes because of morphological similarities with certain fungi (Myxotrichum, Gymnoascus, Arachniotus) belong with them to the Gymnoascaceae. However, this generalization was unwarranted, since their hypothesis was based on a single strain of a dermatophyte, designated by them as Trichophyton, sp. without the species having been more closely determined. Moreover, they did not produce any evidence for their generalization that all dermatophytes should, or necessarily must belong to the Gymnoascaceae. Finally,Matruchot &Dassonville had never seen either pycnidia or perithecia in their cultures. To say the least, their hypothesis, whether right or wrong, must await further demonstrations that pycnidia and/or perithecia do exist in the dermatophytes.The only apparently successful attempt at solving the question of the existence of higher organs of fructification in dermatophytes has been made byNannizzi. His findings must be considered valid, unless duplications of his experiments would prove to the contrary. However, the same objection has to be made against his generalization as was raised against the generalization ofMatruchot &Dassonville. This is that ALL dermatophytes must necessarily belong to the Gymnoascaceae.Nannizzi succeeded in producing pycnidia only in the form-genusTrichophyton, in the speciesTr. mentagrophytes (Ch. Robin, 1853) andTr. equinum Matruchot. He also was successful in his endeavor in the form—genusAchorion, in the speciesAchorion gypseum Bodin. However, his attempts to produce higher organs of fructification inMicrosporon lanosum Sabouraud and inAchorion Quinckeanum Zopf failed.

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Zusammenfassung Versuche, betreffs des gegenwärtigen Standes der Fruchtkörperproduktion in Dermatophyten, die sich nun auf sechs Jahrzehnte erstrecken, sind kritisch beleuchtet worden.Es konnte nur ein einziger, anscheinend erfolgreicher Versuch verzeichnet werden, der vonNannizzi, 1926. Mit seiner speziellen Technik, indem er Dermatophyten auf natürlichen Nährböden (Federn, Leder, Knochen, Haare) züchtete, war es ihm gelungen, Pykniden in der Form-GattungTrichophyton, in den ArtenTr. mentagrophytes (Ch. Robin, 1853) undTr. equinum Matruchot hervorzurufen. In der Form-GattungAchorion sind Pykniden in der ArtAchorion gypseum Bodin produziert worden in einem Nährboden von Waldboden gemischt mit Stückchen von altem Leder. Jedoch sind seine Versuche, Pykniden inMicrosporon lanosum Sabouraud und inAchorion Quinckeanum Zopf hervorzurufen, fehlgeschlagen.

     

A mathematical model of kinetics of the biosynthesis of tetracyclines

Summary A mathematical model simulating the behaviour or Streptomyces aureofaciens in batch culture under conditions when tetracyclines are synthesized in excessive amounts has been formulated. The response of the mathematical model to the experimental conditions applied corresponds with data obtained in the experiments. The mathematical model demonstrated that the level of tetracycline production is determined during the period of culture growth beginning with exhaustion of inorganic phosphate from the medium and ending with inhibition of the synthesis of enzymes caused by the synthesized tetracyclines. Further tetracycline synthesis is then proportional to the amount of enzymes synthesized in this interval.List of symbols E Activity of ACT-oxygenase (10×nkat/g) - P Product concentration (mg/l) - k ₁-k ₆ Rate constants - K _S Saturation constant (g sugar/l) - K _I1 Inhibition constant (mg product/l) - K _I2 Inhibition constant (mM phosphate/l) - K _I3 Inhibition constant (mg product/l) - S ₁ Substrate sucrose (g sugar/l) - S ₂ Substrate concentration — phosphate (mM/l) - r _P Specific rate of product formation (mg product/g · h) - r _E Specific rate of enzyme synthesis (10×nkat/g² · h), Expressed by activity units - t Cultivation time (hour) - X Biomass dry weight (g/l) - Y _S/X Yield coefficient - Specific growth rate (h^-1)     

Analysis of Reproductive Parameters of Urban and Rural Population of Chuvashia

Genetic and demographic characteristics for urban and rural population of the Chuvash Republic (Chuvashes and Russians) were calculated based on 1122 questionnaires. The sibship sizes for Chuvashes were 2.05 (urban) and 2.78 (rural). For Russians these indices were 1.75 (urban) and 2.00 (rural), respectively. Crow’s index and its components were I _m = 0.04; I _f = 0.18; and I _tot = 0.22 for urban, and I _m = 0.07; I _f = 0.27; and I _tot = 0.36 for rural Chuvashes, respectively; and I _m = 0.04; I _f = 0.30; and I _tot = 0.35 for urban, and I _m = 0.03; I _f = 0.29; and I _tot = 0.33 for rural Russians, respectively.__________Translated from Genetika, Vol. 41, No. 6, 2005, pp. 850–854.Original Russian Text Copyright © 2005 by El’chinova, Zinchenko, Kirillov, Abrukova.     

Syntheses of di-hydrocarbyl derivatives of carbonyl phosphine complexes of iron

Complexes cis,trans-Fe(CO)₂(PMe₃)₂RR′ (R = CH₃, R′ = Ph (2); R = CH₃, R′ = CHCH₂ (3); R = CHCH₂, R′ = Ph (4); R = R′ = CHCH₂ (5); R = R′ = CH₃ (6)) were prepared by reaction of cis,trans-Fe(CO)₂(PMe₃)₂RCl (1) with organolithium reagents LiR′. All complexes were characterized in solution by IR and ¹H, ³¹P and, in a few cases, ¹³C NMR mono- and bi-dimensional spectroscopies. Complexes 5 and 6 were structurally characterized by X-ray diffractometric methods. In solution complexes 2, 3 and 4 undergo slowly coupling of the σ-hydrocarbyl substituents leading to Fe(CO)₃(PMe₃)₂ and other decomposition products. Complex 6 was very stable in solution in the absence of nucleophiles and in the solid state. Complex 5 transformed through intramolecular coupling of the vinyl groups into Fe(CO)(PMe₃)₂(η⁴-butadiene) (7), which was characterized in solution by IR and NMR spectroscopies.     

医疗机构合理用药评价模型的构建研究

目的 针对医疗机构的合理用药水平进行评价研究。方法 根据医疗机构合理用药的具体要求,构建医疗机构合理用药评价指标体系,采用基于模糊群决策的方法和多指标评价分析法构建医疗机构合理用药评价模型。结果 构建了基于模糊群决策的医疗机构合理用药评价模型,并通过实例分析证明了评价模型的可行性。结论 建立的基于模糊群决策的医疗机构合理用药评价模型能够对医疗机构的合理用药水平进行科学评价,为提高医疗机构合理用药水平奠定基础。     

Toxic effect of carrageenan on lymphoid-follicle associated epithelial cells of the bursa of Fabricius of chickens

Summary The lymphoid-follicle associated epithelial (FAE) cells of the bursa Fabricii of chickens show enzymatic and micropinocytotic activities; they can perform functions reminiscent of those of macrophages. To test this relationship, we checked whether FAE cells are sensitive to carrageenan, a substance widely known to be toxic for macrophages. This substance was gently introduced into the bursal lumen and was left there for a period of 72 to 120 h. The bursae of Fabricius were then examined histologically and histochemically. The result was that the FAE areas had been reduced in number and that their structural pattern had been modified. This effect suggests that FAE cells may be sensitive to carrageenan. The immunological response to SRBC was studied in one group of animals that had received carrageenan. The antibody seemed to elicit an increase in the agglutinins and in the number of direct PFC/spleen against SRBC in comparison with control chickens. On the basis of the morphological and immunological results obtained, the origin and the role of the FAE cells of the bursa of Fabricius are discussed.

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Résumé Les cellules épithéliales associées aux follicules lymphatiques (FAE) de la bourse de Fabricius du poulet, dotées d'activités enzymatiques et de capacités micropinocitosiques, expriment des fonctions qui, d'une certaine manière, rappellent celles des macrophages. Sur base de cette constatation, on a cherché à évaluer si les cellules FAE sont sensibles à une substance notoirement macrophage-toxique comme la carragénine. Cette substance a été introduite dans le lumière de la bourse de Fabricius où on l'a fait séjourner pendant 72–120. Les bourses de Fabricius prélevées après un tel traitement ont été examinées en employant des méthodes histologiques et histochimiques, qui ont évidencié une réduction des zones FAE avec modification de leur aspect. Les résultats indiqueraient une sensibilité des cellules FAE à la carragénine.Sur un groupe d'animaux traités avec la même substance, on a étudié le comportement immunologique en ce qui concerne SRBC. On observe une hausse des agglutinines et du nombre PFC directes/rate anti-SRBC par rapport aux contrôles non traités avec la carragénine. Sur base des résultats morphologiques et immunologiques, on discute la nature et le rôle des cellules associées aux follicules de la bourse de Fabricius du poulet.

     

The effect of the size of soil structural aggregates on the degree of nitrification

It was experimentally demonstrated that the deviation from linearity exhibited by the dependence of intensity of nitrification on the size of the specific surface of soil structural aggregates is due to deteriorated aeration in the internal space of aggregates of greater diameter. The deteriorated aeration is due to retarded diffusion of oxygen in the aggregates containing greater amounts of moisture. Aggregates isolated from rendzine which displayed a deviation from linearity when their moisture content was equal to twice their maximum hygroscopicity (largersize aggregates produced less nitrates than would correspond to a linear relationship) showed a clear linear dependence as soon as the moisture content dropped to values lower than maximum hygroscopicity.

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Влияние размеров структурных агрегатов почвы на интенсивность нитрификации II. Значение аэрации

Интенсивность нитрификации в почвенных структурных агрегатах различной величины падает с увеличением диаметра агрегатов. Если взять отношение интенсивности нитрификации к размерам специфической поверхности агрегатов (под специфической поверхностью почвенных структурных агрегатов мы понимаем отношение общей внешней поверхности агрегатов опре-деленной фракции к единице вещества), то можно установить, или что падение интенсивности нитрификации протекает равномерно с уменьшением размеров плоцади специфической поверхность агрегатов, или что интенсивность нитри-фикации в агрегатах большего диаметра падает быстрее, чем размеры специфи-ческой поверхности. В этом случае зависимостй интенсивности нитрификации от величины специфической поверхности имеет характер кривой (рис. 1). В предыдущей работе (Сайферт, 1964) нами было высказано предположение, что это явление связано с ухудшением аэрации центральной части крупных агрегатав. Ухудшение аэрации в этих агрегатах, по нашему мнению, обусловлено водяными перегородками, которые здесь почвенная влага образует в результате действия капиллярных сил. Эти перегородки замедляют диффузию кислорода из межагрегатных про-странств внутрь агрегатов. Чтобы доказать правильность нашего предположения, мы устанавливали интенсивность нитрификации в агрегатах раз-личной величины при обычно встречающемся насыщении влагой (19% веса) и при насыщении только до степени гигроскопической влаги. Результаты опытов показали, что в первом случае зависимость интенсивности нитрификации от величины специфической поверхности выражалась кривой (рис. 2), тогда как во бтором случае она имела характер прямой (рис. 4). Ввиду того, что эти результаты полностью подтверждают наше предположение, мы можем считать доказанным, что интенсивность нитрификации в агрегатах боляших диаметров понижается в связи с ухудшением аэрации.

     

Chemistry of the “molecular trap” of protease-catalyzed splicing reaction of complementary segments of α-subunit of hemoglobin A

The complementary fragments of human Hb α, α_1–30, and α_31–141 are spliced together by V8 protease in the presence of 30%n-propanol to generate the full-length molecule (Hb α-semisynthetic reaction). Unlike the other protease-catalyzed protein/peptide splicing reactions of fragment complementing systems, the enzymic condensation of nonassociating segments of Hb α is facilitated by the organic cosolvent induced α-helical conformation of product acting as the “molecular trap” of the splicing reaction. The segments α_24–30 and α_31–40 are the shortest complementary segments that can be spliced by V8 protease. In the present study, the chemistry of the contiguous segment (product) α_24–40 has been manipulated by engineering the amino acid replacements to the positions α²⁷ and α³¹ to delineate the structural basis of the molecular trap. The location of Glu²⁷ and Arg³¹ residues in the contiguous segment α_24–40 (as well as in other larger segments) is ideal to generate (i, i+4) side-chain carboxylate-guanidino interaction in its α-helical conformation. The amino acid residue replacement studies have confirmed that the side chains at α²⁷ and α³¹ facilitate the semisynthetic reaction. The relative influence of the substitute at these sites on the splicing reaction depends on the chemical nature of the side chain and the location. The γ-carboxylate guanidino side-chain interaction appears to contribute up to a maximum of 85% of the thermodynamic stability of the molecular trap. The studies also demonstrate that the thermodynamic stability of the molecular trap is determined by two interdependent conformational aspects of the peptide. One is an amino acid-sequence-specific event that facilitates the induction of an α-helical conformation to the contiguous segment in the presence of organic cosolvent that imparts some amount of protease resistance to Glu³⁰-Arg³¹ peptide bond. The second structural aspect is a site-specific event, ani, i+4 side-chain interaction in the α-helical conformation of the peptide which imparts an additional thermodynamic stability to the molecular trap. The results suggest that conformationally driven “molecular traps” of protease-mediated ligation reactions of peptides could be designed into products to facilitate the modular assembly of peptides/proteins.     

Quantitative and Physiological Analyses of Chloride Dependence of Growth of Halobacillus halophilus

A quantitative analysis of the Cl⁻ dependence of growth of Halobacillus halophilus was performed. Optimal growth rates were obtained at Cl⁻ concentrations of between 0.5 and 2.0 M, and the final yield was also strictly dependent on the Cl⁻ concentration. Br⁻ but not I⁻, SO₄²⁻, NO₂⁻, SO₂⁻, OCN⁻, SCN⁻, BO₂⁻, or BrO₃⁻ could substitute for Cl⁻. To analyze the function of chloride, chloride concentration was determined. At low external Cl⁻ (Cl_e⁻) concentrations, the growth rate was low and Cl⁻ was excluded from the cytoplasm; increasing the Cl_e⁻ concentration led to an increase in the growth rate and an energy-dependent uptake of Cl⁻, thus decreasing the Cl_e⁻/internal Cl_i⁻ gradient from ≥10 at 0.1 M Cl_e⁻ to a nearly constant value of 2 at Cl_e⁻ concentrations which allowed optimal growth. Two membrane proteins with apparent molecular masses of 31 and 16 kDa which were identified to be specific for Cl⁻-grown cultures are possible candidates for a chloride uptake system.