Biological Activity of Fractions Obtained by Differential Centrifugation of the Bovine Adrenal Cortex

This study is an attempt to correlate adrenocortical hormonal activity with various cell fractions obtained by homogenization and differential centrifugation of bovine adrenocortical tissue. Bioassays involving the decrease in circulating eosinophils and sodium retention were employed for the estimation of the glucocorticoids and mineralocorticoids respectively. The supernatant was the most active fraction in both bioassays, while the microsome and mitochondrial fractions demonstrated varying degrees of activity. The total nitrogen, lipide phosphorus, and cholesterol contents of the fractions were determined. The microsome fraction contained a relatively higher amount of lipide phosphorus and a marked amount of cholesterol.     

Morphological fractions, chemical composition and in vitro fermentation characteristics of maize stover of five genotypes

Five morphological fractions (leaf blade, leaf sheath, stem, husk and cob) of stover of five maize genotypes, namely waxy, conventional, fodder, sweet and high-oil maize, respectively, were used to test the effects of genotype and morphological fractions on chemical composition and in vitro fermentation characteristics. The waxy maize had a higher (P < 0.05) stem but lower (P < 0.05) leaf blade proportion and fodder maize had a higher (P < 0.05) leaf blade but lower (P < 0.05) leaf sheath proportion than other genotypes, respectively. Maize genotype had a significant effect (P < 0.001) on the chemical composition of stover parts except for organic matter (OM) concentration. Chemical composition of stover parts was affected (P < 0.001) by morphological fractions. The interaction effects between genotype and morphological fraction on the fiber content of stover parts were significant. Over 0.40 and 0.50 of phosphorus (P) and crude protein (CP) of whole-plant maize stover were averagely contributed by leaf blade. Leaf blade, stem and cob contributed over 0.75 of OM, CP, P and fiber in the whole plant. There were significant effects of genotype and morphological fraction on both in vitro gas production parameters and in vitro organic matter disappearance of maize stovers. The genotype and morphological fraction of maize stover and their interaction had significant effects on NH3-N and total volatile fatty acid concentration and the molar proportion of volatile fatty acid in the supernatant after 72 h of incubation except for valeric acid. The present data indicated that the genotype and morphological fraction of maize resulted in variation in the nutritive value of maize stover.     

PHOSPHATE METABOLISM IN BLUE-GREEN ALGAE. II. CHANGES IN PHOSPHATE DISTRIBUTION DURING STARVATION AND THE “POLYPHOSPHATE OVERPLUS” PHENOMENON IN PLECTONEMA BORYANUM

Physiological aspects of phosphate utilization by the blue-green alga Plectonema boryanum were studied. It was found that the external phosphate concentration influenced the distribution of phosphorus-containing compounds in the cell. Culturing the alga in concentrations of 10, 100, and 1000 mg PO₄/l resulted in increases in the level of acid-soluble and acid-insoluble polyphosphates. The values reported for 100 and 1000 mg PO₄/l were the same, indicating that the cells were able to assimilate and utilize only fixed amounts of phosphates. The total phosphorus value for these cells was calculated to be 6.5 μg P per 10⁶ cells. Culturing the alga in 1 mg PO₄/l led to a decrease in phosphate concentration of all cell fractions. Cells grown in the absence of phosphate for 5 days had total cell phosphorus levels of 0.76 μg P per 10⁶ cells. Cells in culture for two months or longer were found to have total cell phosphorus levels of 0.73 μg P per 10⁶ cells. This was determined to be the minimum cell phosphorus level limiting growth. Transfer of cells from either culture condition to a medium containing phosphate led to an “overplus” phenomenon. This overplus phenomenon was characterized by increases in all cellular phosphorus fractions. The most dramatic increase was found in both the acid-soluble and acid-insoluble polyphosphates. These fractions often increased by more than an order of magnitude. The greatest phosphate uptake occurred within 1 hr of transfer of phosphate-starved cells into a medium containing a known amount of phosphate and is essentially complete at 4 hr. The total cell phosphorus levels for uptake never increased beyond 18.9 μg per 10⁶ cells.     

Incorporation of phosphorus into phased cells ofCandida utilis using32P and33P

Changes in phosphorus metabolism were studied by examining the incorporation of³²P and³³P into cells ofCandida utilis growing in phased culture during a 6 h cell cycle and a post-cycle period of 6 h. Three different chemically defined media were used; these were phosphorus, nitrogen and carbon limited. The patterns of incorporation of phosphorus into RNA, DNA, lipid and cold water extractable phosphate fractions showed a non-uniform behaviour during both cell cycle and post-cycle periods. The patterns were different in all three types of media. The results showed that a cell can grow and develop at a fixed growth rate in different ways: so that the pattern of behaviour during a cell cycle is not stereotyped for a given doubling time, but largely depends upon the nutrient environment in which the cell exists.     

The phosphorus fractions and alkaline phosphatase activities in sludge

The alkaline phosphatase activities (APA) and phosphorus fractions in activated sludge during wastewater treatment were studied. Our results showed that the phosphorus concentration and fractions in activated sludge were highly correlated with the characteristics of influents. Inorganic phosphorus (IP) and non-apatite inorganic phosphorus (NAIP) were the main phosphorus fractions of sludge. A larger phosphorus concentration was found in activated sludge due to the more readily mobilizable and bio-available forms. The APA in sludge was directly correlated with mixed liquor suspended solids (MLSS) in activated sludge. The APA in the sludge is implicated the depletion of organic phosphorus forms in sludge, whilst also implying its less inhibition of inorganic phosphorus in sludge. The APA and phosphorus fractions in different sludge samples from the same wastewater treatment plant were quite stable. This stability shows their tight interactions in sludge.     

Plasma membrane fragments in bovine and caprine skim milks

By either differential or linear gradient ultracentrifugation of bovine or caprine skim milks it was possible to obtain fractions which contained 45–75% of the lipid phosphorus and unesterified cholesterol of the skim milk. Electron microscopy of these fractions revealed the presence of numerous membrane-bound vesicles, microvilli and membrane fragments. Assay of the fractions for certain membrane-bound enzymes; i.e. 5′-nucleotide pyrophosphatase, alkaline phosphatase and ATPases, established the presence of all but the latter in the membrane-rich fractions. The distributions of the enzymes in the various fractions were correlated with their lipid phosphorus and cholesterol contents.Compositions of the phospholipids from skim milk membranes, milk fat globule membranes and the plasma membrane of the lactating mammary cell were observed to be similar and unique for having a relatively high level (20–25%) of sphingomyelin. By virtue of secretory processes, all of these membranes appear to be interrelated with each other and with Golgi vesicle membranes. It is concluded that the membrane material in the skim milk originates primarily from plasma membrane of the lactating cell. The possibiltiy that Golgi vesicle membranes form a substantial part of this material is not precluded by the results of this study.Separation of bovine skim milk on a Sepharose 4B gel column demonstrated that virtually all of the 5′-nucleotidase and lipid phosphorus are recovered together in the void volume of the column. Considering the particle size discriminating characteristics of this gel, the skim milk membrane material appears to be constituted of relatively large structures rather than of discrete subunits.     

普通小球藻生长与武汉东湖水体磷形态的相关研究

本文探讨了武汉东湖水体中总溶解磷（TSP）、溶解反应磷（SRP）、总反应磷（TRP）、溶解水解磷（SHP）和颗粒磷（PP）对普通小球藻生长的生物有效利用性。进行了普通小球藻生长量与各磷形态及总磷（TP）被利用浓度的一元相关分析,进一步讨论了这些一元相关方程的不足,并用多元回归分析建立了普通小球藻生长量（N）与所测几种磷形态被利用浓度（CTSP,CSRP,CTRP,CSHP,CPP）的相关模型。这些模型可以用于评价湖泊水体富营养化的程度及预测藻的生长潜力。     

Successive changes in the blood composition of experimental normal beagle dogs associated with age

Eight hematologic parameter values, 16 serum biochemical constituents, serum protein fractions and albumin-globulin ratios were determined in blood samples obtained from 879 normal, healthy Beagle dogs of both sexes which had been reproduced and bred in our laboratories. The blood samples were collected from the Beagles that ranged in monthly ages from 1 to 12 and in monthly ages from 13 to 121, which were classified as the adult class. As a result, red blood cell counts, hemoglobin concentrations and packed cell volumes increased with growth. Red blood cell parameters of normal Beagles in our laboratories were rather higher than those in literatures presented by many other researchers. MCV decreased and MCHC increased gradually with age. Total serum protein concentrations increased with growth. alpha 1-1 and alpha 1-2 Globulin fractions descended, but beta 2 and gamma globulin fractions ascended in serum proteins. Alkaline phosphatase activities, inorganic phosphorus concentrations and glucose concentrations decreased conspicuously with growth. Leucine aminopeptidase activities and calcium concentrations decreased slightly. Serum cholinesterase and LDH activities showed a tendency to diminish similarly. Blood urea nitrogen and creatinine concentrations multiplied gradually. Hematologic parameters became almost steady in our 7-month-old dogs or older ones and serum biochemical constituents had a tendency to be stable in our 7- to 9-month-old dogs or older ones in the blood. White blood cell counts, alkaline phosphatase activities, inorganic phosphorus concentrations, glucose concentrations, leucine aminopeptidase activities and calcium concentrations were lowest in the adult class.     

华西雨屏区不同林龄柳杉人工林土壤磷组分特征

磷是限制陆地生态系统生产力的关键养分因子。当前尚不了解不同土壤磷组分随柳杉林龄增长如何变化,及其与土壤微生物群落的关系。以华西雨屏区不同林龄（7年生幼龄林、13年生中龄林、24年生近熟林、33年生成熟林,53年生过熟林）柳杉（Cryptomeria japonica var. sinensis）人工林为研究对象,采用Hedley磷素分级方法、磷脂脂肪酸分析法（PLFA）来探究不同林龄柳杉人工林土壤磷组分的分布模式及影响因子。结果表明：不同林龄和土壤深度土壤磷各组分含量差异显著。随林龄增加,可溶性磷和磷灰石含量逐渐减少,残余态磷含量逐渐增加,其余磷组分含量先增加后降低。除可溶性磷、浓盐酸提取态无机磷和残余态磷组分外,其余土壤磷组分含量表现为上层（0-15 cm）高于下层（15-30 cm）。偏门特尔检验表明,微生物群落与土壤磷组分之间存在显著关联。回归分析发现,碳与有机磷比值和酸性磷酸酶活性呈正相关。冗余分析显示,pH、土壤有机碳、土壤含水量,全氮和土壤容重是影响土壤磷组分变化的主导因子。研究显示,造林初期的土壤磷组分快速积累,在中龄林阶段达到最大值,随着柳杉人工林林龄的增加,土壤中磷的限制逐渐加强,土壤磷组分含量在成熟林之后逐渐下降。这些结果可为为柳杉人工林的培育及可持续经营管理提供科学依据。     

Longitudinal Variability of Phosphorus Fractions in Sediments of a Canyon Reservoir Due to Cascade Dam Construction: A Case Study in Lancang River,China

Dam construction causes the accumulation of phosphorus in the sediments of reservoirs and increases the release rate of internal phosphorus (P) loading. This study investigated the longitudinal variability of phosphorus fractions in sediments and the relationship between the contents of phosphorus fractions and its influencing factors of the Manwan Reservoir, Lancang River, Yunnan Province, China. Five sedimentary phosphorus fractions were quantified separately: loosely bound P (ex-P); reductant soluble P (BD-P); metal oxide-bound P (NaOH-P); calcium-bound P (HCl-P), and residual-P. The results showed that the total phosphorus contents ranged from 623 to 899 µg/g and were correlated positively with iron content in the sediments of the reservoir. The rank order of P fractions in sediments of the mainstream was HCl-P>NaOH-P>residual-P>BD-P>ex-P, while it was residual-P>HCl-P>NaOH-P>BD-P>ex-P in those of the tributaries. The contents of bio-available phosphorus in the tributaries, including ex-P, BD-P and NaOH-P, were significantly lower than those in the mainstream. The contents of ex-P, BD-P, NaOH-P showed a similar increasing trend from the tail to the head of the Manwan Reservoir, which contributed to the relatively higher content of bio-available phosphorus, and represents a high bio-available phosphorus releasing risk within a distance of 10 km from Manwan Dam. Correlation and redundancy analyses showed that distance to Manwan Dam and the silt/clay fraction of sediments were related closely to the spatial variation of bio-available phosphorus.     

Lipids of human leukocytes: relation to celltype

Significant differences in lipid composition have been found between normal human lymphocytes and polymorphonuclear leukocytes (isolated from blood by means of glass-bead columns), abnormal leukocytes from patients with acute and chronic leukemia, and leukocytes from peritoneal exudates. Lipid extracts of isolated leukocytes were analyzed for total lipid, phosphorus, cholesterol, and plasmalogens. Individual phospholipids and neutral lipids were separated by thin-layer chromatography. The major phospholipids were phosphatidyl choline, ethanolamine glycerophosphatides, sphingomyelin, phosphatidyl serine, and phosphatidyl inositol. Plasmalogen was found mainly as phosphatidal ethanolamine. The neutral lipid fractions contained free cholesterol and various amounts of triglyceride, but little esterified cholesterol. Normal lymphocytes contained about half as much total lipid per cell as normal polymorphonuclear leukocytes, with a similar cholesterol:-lipid-P ratio but relatively more lecithin and less ethanolamine glycerophosphatide. Normal mature leukocytes, compared with immature cells of the same morphological series, had a higher total lipid content per cell, more cholesterol, and a higher ratio of cholesterol to lipid-P. Little difference was found in total lipid-P per cell, but mature cells contained relatively less lecithin and more sphingomyelin. These findings may reflect differences in the relative content of various intracellular organelles as well as possible differences in the quantity and composition of the plasma membrane.     

Intracellular distribution of 5' nucleotidase in rat liver

The intracellular distribution of 5' nucleotidase was investigated in rat liver by biochemical analysis of cell fractions obtained by differential centrifugation. The enzymatic activity was measured by determination of the inorganic phosphorus liberated from 5' nucleotides. The 5' nucleotidase activity was mainly found in the nuclear and microsomal fractions. An attempt to extract the enzyme from these fractions with Mg(++) ion solutions was unsuccessful. It is concluded that 5' nucleotidase is actually present in the nuclear and microsomal fractions of rat liver cells.     

Effect of Phosphorus Deficiency on Carbohydrate Metabolism of Mentha arvensis

The effects of phosphorus deficiency on carbohydrate fractions of Mentha arvensis L. var. piperascens were studied. Mint plants were grown in sand cultures under full nutrition and phosphorus deficiency conditions. Component organs, viz., leaf, stem and root, were sampled at four different stages of the growth cycle and analysed for various sugar fractions. In leaf and root phosphorus deficiency brought about a definite increase in all sugar fractions whereas in stem a reduction was noticed. The stem appeared to be the principal storage organ throughout the growth. Maximum sugar concentration was recorded at the age of 70 days, which represents the full maturity stage, and coincided with maximum essential oil accumulation.     

Intracellular Distribution of 5' Nucleotidase in Rat Liver

The intracellular distribution of 5' nucleotidase was investigated in rat liver by biochemical analysis of cell fractions obtained by differential centrifugation. The enzymatic activity was measured by determination of the inorganic phosphorus liberated from 5' nucleotides. The 5' nucleotidase activity was mainly found in the nuclear and microsomal fractions. An attempt to extract the enzyme from these fractions with Mg⁺⁺ ion solutions was unsuccessful. It is concluded that 5' nucleotidase is actually present in the nuclear and microsomal fractions of rat liver cells.     

Variation in polar-group content in lipids of cowpea (Vigna unguiculata) Cell cultures as a mechanism of haloadaptation

Callus and cell suspension cultures of cowpea (Vigna unguiculata) were induced with 2,4-dichlorophenoxyacetic acid and grown at different NaCl concentrations. The cell biomass yield and its total lipid content decreased with increasing salinity. However, while the hexose content in lipids was higher, the amount of lipid phosphorus was significantly lower in both agar and cell suspension cultures. Ion-transport rates with artificial membranes prepared with different lipid fractions showed that lipids from cells grown in a saline medium were less permeable to Na⁺ and to Cl^- than those grown in a non-saline medium. Also the permeability of membranes prepared with glycolipids was lower than those prepared with phospholipids and whole lipids. Apparently, the increase of hexose/phosphorus ratio in membrane lipids is induced in response to the halo-adaptation process.     

长期施肥对黄土旱塬农田土壤有机磷组分及小麦产量的影响

研究长期施肥对黄土旱塬农田土壤有机磷组分及小麦产量的影响,可为提高磷素转化利用率及合理利用肥料提供理论支持。本研究依托长武旱塬农田生态系统长期(1984—2016年)定位试验站,选取不施肥(CK)、单施氮肥(N)、单施磷肥(P)、施氮磷肥(NP)、单施有机肥(M)、氮肥配施有机肥(MN)、磷肥配施有机肥(MP)、氮磷肥配施有机肥(MNP)8个处理,研究其对土壤有机磷组分、小麦产量和土壤性质的影响。结果表明: 长期施肥后土壤有机磷含量为244.7～429.1 mg·kg^-1,除N处理外,其余各处理有机磷含量比CK均显著增加了15.4%～47.9%。长期施用磷肥改变了黄土旱塬农田表层土壤(0～20 cm)各有机磷组分含量,MP、MNP处理显著提高了活性有机磷及中活性有机磷含量;N、P和NP处理显著降低了中稳性有机磷含量;N、P、NP、MN、MP、MNP处理均显著提高了高稳性有机磷含量。各处理土壤有机磷组分与总有机磷含量比值为:中活性有机磷>高稳性有机磷>活性有机磷>中稳性有机磷。长期施肥后,与CK相比,氮、磷肥配施,尤其是与有机肥配施,显著增加了小麦生物产量和籽粒产量。土壤指标中,有机质、速效磷和无机磷与小麦产量呈显著正相关。MP、M处理可以显著提高黄土旱塬黑垆土中的速效磷、总磷、总无机磷、活性有机磷和中活性有机磷含量,表明有机肥与磷肥配施可以提高该地区更容易被作物吸收的磷组分。总之,氮磷肥配施并配施有机肥可以提高该地区磷供给,对小麦增产有促进作用,对提高黄土旱塬地区土壤质量有重要意义。     

Ethylene and phosphorus availability have interacting yet distinct effects on root hair development

The hypothesis that ethylene participates in the regulation of root hair development by phosphorus availability in Arabidopsis thaliana was tested by chemically manipulating ethylene synthesis and response and with ethylene-insensitive mutants. Low phosphorus-induced root hair development could be mimicked by adding the ethylene precursor, 1-aminocyclopropane-1-carboxylate (ACC), to high phosphorus media, and inhibited by adding ethylene inhibitors to low phosphorus media. Ethylene-insensitive mutants showed a reduced response to low phosphorus, indicating ethylene involvement in root hair responses to phosphorus deficiency. To dissect the nature of this involvement, the morphological and anatomical changes associated with increased root hair density were investigated. Growth in low phosphorus resulted in smaller, more numerous cortical cells, resulting in a larger number of root hair-bearing epidermal cell files. Cortical cell number was not affected by ethylene inhibitors, ACC, or mutations reducing ethylene sensitivity in roots grown with low phosphorus, indicating that ethylene does not participate in this response. The exception was the eir1 mutation, which strongly reduced this change in radial anatomy, supporting a role for polar auxin transport in this process. Trichoblast cell length was reduced by low phosphorus availability in all genotypes, but even more so for ein2-1 and ein4. The proportion of epidermal cells forming hairs and root hair length were reduced in ethylene-insensitive mutants, especially in the presence of low phosphorus. These results demonstrate multiple effects of low phosphorus from the earliest stages of root hair development, and cross-talk between ethylene and phosphorus in the control of a subset of the low phosphorus effects, concentrating on those later in development.     

Biochemical and immunological properties of Coxiella burnetii cell wall and peptidoglycan-protein complex fractions.

Coxiella burnetii morphological cell types were fractionated into large-cell variant cell walls, two fractions of small-cell variant cell walls, and one fraction of small-cell variant whole cells. Based on the contents of peptidoglycan (PG)-constituents and the yields of the sodium dodecyl sulfate-insoluble PG-protein complex (PG-PC) from cell walls, the fraction of large-cell variant cell walls contained significantly less PG than did the fraction of small-cell variant cell walls. The yields of PG-PC from the fractions of large-cell variant cell walls and small-cell variant cell walls were 2 and 32%, respectively. These results indicated that the PG of the large-cell variant cell walls may be partially digested by PG-lytic enzymes or incompletely synthesized, whereas the small-cell variant cell walls appeared to have intact PG. Proteins associated with PG-PC were resistant to proteolysis by trypsin, protease VI, and proteinase K. Saturated and unsaturated fatty acids were detected in whole cells and cell walls but not in PG-PC, which contained a 3-deoxy-D-mannooctulosonic acid-like component that is also present in phase I lipopolysaccharide. Immunogenicity of the fractions was tested by measuring the temporal sequence of phase II and phase I antibody responses in vaccinated rabbits. Both phase II and phase I antibody responses were demonstrated with all fractions except the sodium dodecyl sulfate supernatant of the small-cell variant cell walls, whereas PG-PC elicited a pure phase II antibody response up to 29 days postvaccination. The immunogenicity of these fractions may reflect a quantitative difference in antigen concentration or may be due to a qualitative difference in phase II and I determinants.     

Characterization of degradation products of the cell wall released during growth and sporulation ofBacillus megaterium

Asporogenic and sporogenic strains ofBacillus megaterium KM release during growth heterogeneous fragments of the cell wall into the medium the non-dialyzable fraction representing 50–90% by the total. During lysis of sporangia the non-dialyzable fraction represents only 30% of the soluble fraction of autolyzed walls. Gel filtration on Sephadex permits to separate the non-dialyzable fragments of the cell wall released during growth into two fractions contaning simultaneously peptidoglycan and phosphorus. The two fractions contain peptidoglycan components in the same ratio as in the cell wall. Only one peptidoglycan macromolecular fraction, smaller than the fractions released during growth, was detected by gel filtration in the material released during lysis of sporangia.     

Primary distribution of n-alkane through the structures of yeast cells]

When n-alkane enters the cells as a result of molecular sorption and diffusion, it is distributed within all membrane structures of the cell (microsomes, mitochondria, membranes, vacuoles, cytoplasmic membranes, and cell walls). Sorption of n-alkane by all these structures in vivo comes at equilibrium after incubation of the cells with n-alkane during 3-4 min. Accumulation of the hydrocarbon in the morphological fractions of the cell depends on its concentration in the incubation medium. Isotherms of the sorption are convex curves. Sorption power, maximum sorption capacity, affinity and strength of the bonds with the hydrocarbon differ among the membrane structures of the cell. The maximum capacity of sorption of n-alkane by the structures does not correlate with the content of lipids and phospholipids in the structures. Sorption of n-alkanes is presumed to depend on the structural organization of lipids in the morphological fractions.