Partial purification,subunit structure and thermal stability of the photochemical reaction center of the thermophilic green bacterium Chloroflexus aurantiacus

Spectrally pure reaction center preparations from Chloroflexus aurantiacus have been obtained in a stable form; however, the product contained several contaminating polypeptides. The reaction center pigment molecules (probably three bacteriochlorophyll a and three bacteriopheophytin a molecules) are associated with two polypeptides (M_r = 30000 and 28000) in a reaction center complex of M_r = 52000. No carotenoid is present in the complex. These data together with previous spectral data suggest that the Chloroflexus reaction center represents a more primitive evolutionary form of the purple bacterial reaction center, and that it has little if any relationship to the green bacterial component. A reaction center preparation from Rhodopseudomonas sphaeroides R26 was fully denatured at 50°C while the Chloroflexus reaction center required higher temperatures (70–75°C) for complete denaturation. Thus, an intrinsic membrane protein of a photosynthetic thermophile has been demonstrated to have greater thermal stability than the equivalent component of a mesophile.     

Inactivation of Rhodospirillum rubrum coupling factor by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole. Modification of a tyrosine protected by phosphate

Chemical modification of Rhodospirillum rubrum chromatophores by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) results in inactivation of photophosphorylation, Mg²⁺-ATPase, oxidative phosphorylation and ATP-driven transhydrogenase, with apparent first-order kinetics. Other energy-linked reactions such as light-driven transhydrogenase and light-dependent proton uptake were insensitive to NBD-Cl. The Ca²⁺-ATPase activity of the soluble coupling factor from chromatophores (R. rubrum F₁) was inactivated by NBD-Cl with kinetics resembling those described for Mg²⁺-ATPase and photophosphorylation activities of chromatophores. Both NBD-chromatophores and NBD-R. rubrum F₁ fully recovered their activities when subjected to thiolysis by dithioerythritol. Phosphoryl transfer reactions of chromatophores and Ca²⁺-ATPase activity of R. rubrum F₁ were fully protected by 5 mM P_i against modification by NBD-Cl. ADP or ATP afforded partial protection. Analysis of the protection of Ca²⁺-ATPase activity by P_i indicated that NBD-Cl and P_i are mutually exclusive ligands. Spectroscopic studies revealed that tyrosine and sulfhydryl residues in R. rubrum F₁ underwent modification by NBD-Cl. However, the inactivation was only related to the modification of tyrosine groups.     

PHOTOSYNTHETIC QUANTUM EFFICIENCIES OF PHYTOPLANKTON FROM PERENNIALLY ICE COVERED ANTARCTIC LAKES1

A recently introduced approach far estimating the photosynthetic quantum efficiency (φ) of a freshwater or marine phytoplankton community has been applied for the first time to high latitude polar ecosystems, namely four lakes of southern Victoria Land, Antarctica. Values for φ at various depths ranged from 0.0022–0.1560 when calculated using a recommended mean extinction coefficient for phytoplankton (i.e. k?_c= 0.016). By contrast, φ ranged from 0.0037–0.0760 when calculated using an empirically estimated value for k?_c of 0.0328. If the recommended k?_c= 0.016 more closely approaches an accurate estimate, then the φ valves indicate that the phytoplankton convert light to organic carbon more efficiently than elsewhere. However, if the empirically derived k?_c= 0.0328 more closely approaches an accurate estimate, then the φ values indicate the phytoplankton trap light more efficiently than elsewhere. Although we have not resolved whether light conversion (φ) or light trapping are more efficient, the results show that the phytoplankton of these Antarctic lakes are well adapted to performing photosynthesis under extremely low light conditions.     

Photosynthetic characteristics of leaves developed at different irradiances and temperatures: an extension of the current hypothesis

Photosynthetic characteristics at high measurement irradiance were analyzed for single leaves of two C₃ and one C₄ species grown under twenty one combinations of irradiance level, irradiance duration, and air temperature in order to test the idea that photosynthetic characteristies developed by leaves in different environments are controlled by the daily amount of photosynthesis. Photosynthetic rates per unit area and mesophyll conductances at 25°C and air levels of CO₂ and O₂, and parameters for two photosynthesis models were used to characterize the photosynthetic properties of the leaves. Leaves with highest values of the photosynthetic parameters for each species were often developed in environments with irradiance levels below saturation for photosynthesis, and with only 12 hours of irradiance per day. Lower air temperature during growth increased the photosynthetic characteristics for a given irradiance regime. Photosynthetic characteristics had higher correlation coefficients with daily photosynthesis of mature leaves divided by 24-hour leaf elongation rates of young leaves, than with daily photosynthesis alone, indicating that photosynthetic characteristics may be related to a balance between photosynthesis and leaf expansion.     

Mode de formation des inclusions lamellaires dans le poumon embryonnaire de poulet

Résumé Les pneumocytes granuleux, qui constituent l'un des principaux types cellulaires de l'épithélium pulmonaire, sont caractérisés par la présence de volumineuses inclusions osmiophiles lamellaires.Nous avons étudié l'apparition et l'origine de ces inclusions dans l'épithélium du poumon embryonnaire de Poulet, en l'examinant à différents stades du développement.Les premières inclusions lamellaires apparaissent dans le poumon de l'embryon de 16 jours. A ce stade, quelques lamelles concentriques entourent une zône centrale amorphe étendue; la périphérie des inclusions contient toujours de petites structures granulaires. Les jours suivants le nombre de cellules contenant des inclusions lamellaires augmente rapidement; en même temps, les lamelles deviennent plus nombreuses. A 19 jours, les inclusions lamellaires ont un aspect semblable à celui qu'elles ont dans les poumons d'animaux adultes.Dès l'apparition des ébauches pulmonaires, à 2 1/2 jours d'incubation, les cellules épithéliales contiennent des inclusions typiques: les inclusions granulaires. Ces organites sont caractérisés par un centre granulaire, qu'entouré un système membranaire. Ce système, simple chez le jeune embryon, évolue ensuite en se compliquant; chez l'embryon de 16 jours, il s'enroule en plusieurs couches autour de la masse centrale. Au moment où les premières inclusions lamellaires apparaissent, le nombre des inclusions granulaires augmente rapidement; on les trouve souvent étroitement associées à des vacuoles lipidiques.L'analyse des relations entre inclusions lamellaires, inclusions granulaires et vacuoles lipidiques suggère que l'inclusion lamellaire résulte de la collaboration entre une vacuole lipidique et plusieurs inclusions granulaires.

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Differentiation of lamellar inclusions in the chick embryonic lung

Summary The granular pneumocytes, one of the main cellular types of the lung epithelium, are characterized by the presence of large osmiophilic lamellar inclusions. The appearance and origin of these inclusions has been studied in the epithelium of chick embryonic lung at different developmental stages.Lamellar inclusions are first seen in the lung of 16 day old embryos. At this stage, few concentric lamellae surround a large amorphous center; the periphery of the inclusions always contains small granular structures. In the following days, the number of cells containing these lamellar inclusions increases rapidly, while their lamellae progressively become more numerous. In 19 day old embryos, the lamellar inclusions are similar to those in the lungs of adult animals.From the earliest formation of the bronchial primordia, their epithelial cells contain a number of typical granular inclusions. These organelles are characterized by a granular center, enclosed in a membranous system. This structure becomes more complex as the embryo develops; in the 16 day old embryo, the multilayered membranous system coils around the granular center. At the time when lamellar inclusions first appear, granular inclusions increase rapidly in number and are often found in close association with lipidic vacuoles.The relationships between lamellar inclusions, granular inclusions and lipidic vacuoles are discussed. The evidence suggests that a lamellar inclusion arises from the cooperation of several granular inclusions and a lipidic vacuole.

     

Pregnant mares' serum gonadotropin source influences fertilization and fresh or thawed embryo development,but the effect is genotype specific

The influence of the source of pregnant mares' serum gonadotropin (PMSG) on the num ber, quality, and in vitro development of mouse embryos before and after freezing was evaluated among three genotypes: N:NIH(S), C57BL/6N, and C3H/HeN-MTV^?. Immature females were given PMSG from one of five commercial sources. Following col lection ( 116 hr later), embryos were evaluated for stage of development, and four-to eight-cell embryos were pooled within genotype and assigned to standardized fresh or freeze-thaw culture trials. Different PMSG sources stimulated the production of different num bers of total embryos (P < 0.05) but not necessarily more embryos suitable for freezing. Differences in embryo production among genotypes indicated that absolute embryo num bers using a single mouse genotype may not accurately reflect the potency of a specific gonadotropin source. The PMSG source also affected the ability of an embryo to survive in culture either immediately after collection or after frozen storage. The effect, however, was genotype specific, with some mouse strains being relatively insensitive to PMSG source, whereas gonadotropin source played a major role in determining in vitro viability in others. Development rates for freshly collected embryos differed, often inconsistently, from those of thawed embryos regardless of the PMSG source used, demonstrating that fresh embryo development cannot be used to estimate expected post-thaw survival. In vitro development of thawed embryos is influenced not only by genotype, but also the source of the gonadotropin used to promote follicular development and oocyte maturation. These findings may explain, in part, the wide variation in embryo viability and culture rates reported among laboratories and intraspecies animal populations.     

Direct effects of serotonin and ketanserin on the functional morphology of embryonic chick skin in vitro

Summary Different organotypical culture methods are used to test the direct effects of serotonin and ketanserin, a S₂, α₁, and H₁ receptor antagonist in vascular tissue, on fibroblasts and epidermal cells of embryonic chick skin in vitro. From light microscopic and electron microscopic analyses, we learn that serotonin enhances keratinization and differentiation, whereas ketanserin reduces differentiation in comparison to the control cultures. Incorporation data of fragments cultured with [³H]thymidine show that ketanserin, within a dose range from 0.05 to 5 μg/ml, stimulates proliferation. Serotonin at a concentration of 10 μg/ml slightly slows down proliferation, whereas lower doses of 0.1 and 1 μg/ml result in tritium activities that do not differ from control cultures. This investigation was financially supported by the National Fund of Scientific Research, Belgium, 3.0022.87.     

Further purification of a fibroblast growth factor-like factor from chick embryo extract by heparin-affinity chromatography

Summary A mitogenic factor which promotes quail myoblast proliferation has been purified some 10⁵-fold from chick embryo extract by a combination of cation-exchange chromatography and heparin-affinity chromatography. The factor is eluted from heparin-Sepharose with 2M NaCl and is a single-chain polypeptide with an apparent molecular weight of 15000 to 17000. It is active at subnanogram level in triggering the proliferation and thereby delaying temporarily fusion of myoblasts. It also stimulates the proliferation of quail fibroblasts in a similar effective concentration range. For both myoblasts and fibroblasts the dose-response to the factor is quantitatively and qualitatively comparable with that of bovine pituitary fibroblast growth factor. These observations strongly suggest that the factor very probably corresponds to chicken fibroblast growth factor or to a closely related molecule(s) and that it is possibly involved in the regulation of myogenesis. This work was partly supported by a grant from the National Center of Neurology and Psychiatry (NCNP grant 86-01) of the Ministry of Health and Welfare, Japan.     

注射依赖cAMP的蛋白激酶催化亚基加速胚胎内细胞间连接通讯的发育(英文)

本研究以爪蟾胚胎内胚层细胞间连接通讯发育的时程为指标,观察了cAMP和依赖cAMP的蛋白激酶催化亚基对这一发育的影响。将依赖cAMP的蛋白激酶催化亚基注射入爪蟾胚胎四细胞期的每一个细胞可使该胚胎内胚层细胞间连接通讯的发育明显加快,提示在正常状态下这一发育的进程是受细胞内依赖cAMP的磷酸化水平的制约。但用双丁酰cAMP和磷酸二酯酶抑制剂对胚胎进行培育,或将cAMP和磷酸二酯酶抑制剂注射入胚胎细胞,对这一发育并无影响,可能是由于这些胚胎细胞内依赖cAMP的蛋白激酶的实际有效含量较低而cAMP含量较高所致。注射这种蛋白激酶催化亚基所诱导的胚胎细胞间连接通讯在注射后约8.5小时出现。这一时间比前人在哺乳动物细胞培养中用cAMP或必需的信使RNA诱导出细胞间连接通讯所需要的时间(2—4小时)长得多,提示在胚胎中依赖cAMP的磷酸化对细胞间连接通讯发育的作用是从RNA转录的水平上开始的。     

基因型和胚龄对小麦未成熟胚离体培养反应的影响

本文对34种基因型的小麦未成熟胚在离体培养中的反应进行了比较。结果表明,94％的供试基因型愈伤组织诱导率都可达到80％以上,若排除供体植株环境条件的不同和接种过程中的人为因素可能造成的影响,不同基因型的愈伤组织诱导率看来没有根本的差异。愈伤组织分化率因基因型的不同变动在0—60％之间,平均为32.7％。虽然同一基因型的盾片愈伤组织分化率在不同年份中有所不同,但是愈伤组织是否具有再生能力?看来是个稳定的遗传性状。因此小麦未成熟胚对愈伤组织诱导的反应和愈伤组织的再生能力可能具有不同的遗传基础。本文的结果还表明,虽然最适于培养的未成熟胚的大小为1毫米左右,伹小至0.3毫米的未成熱胚仍能以几乎100％的频率形成愈伤组织,60％左右的愈伤组织能分化出再生檀株,只是所需的时间比1毫米左右的胚较长。