Some Aspects of Translocation in Root Nodule Plants

By the use of ¹⁶N it is shown that the removal, over a shortzone at the base of the shoot of a typical root nodule plant(alder), of the tissues external to the xylem does not interferewith the upward movement of fixed nitrogen from the nodulesinto the shoot. It is concluded that the fixed nitrogen, whichis probably in organic form, can be translocated in the xylemin the transpiration stream, and that this is most likely itsnormal route. It is also shown that in unringed plants substantialenrichment in fixed ¹⁶N is detectable in the shoot within 6hours from the commencement of the exposure of the nodules toexcess free ¹⁶N. Structural and experimental evidence showsthat the alder nodule is not a water-absorbing organ, and themechanism of transfer of fixed nitrogen from the nodule intothe transpiration stream is not obvious.     

A Quantitative Investigation of a Peristaltic Model for Phloem Translocation

An analysis is made of a peristaltic model of phloem translocationIt is postulated that the periodic action of contractile orbending organelles drive a longitudinal flow of solution withintubules which connect sieve pores in successive sieve plates Plausible values are assumed for the velocity of propagationof the contraction wave, the frequency, the amplitude, and theviscosity of the solution and its concentration. Using relationswhich describe peristaltic flow, predictions are made for thevalues of parameters such as the velocity of solution, the drivingpressure, and rate of energy dissipation. These predicted valuesare seen to be reasonable when compared with the known propertiesof other biological contractile systems. Thus the model is quantitativelyacceptable.     

Quantitative Aspects of Insect Nutrition

The small size of insects, correlated with high food-consumption,growth, and metabolic rates, makes possible their high efficiencyof food-utilization, short life cycles, high biotic potential,and rapid genetic adaptation and speciation. Most of the qualitativenutritional requirements are known for many insect species;many essential nutrients are provided by microbial symbiotes.Recent studies in quantitative nutrition make possible moredetailed analysis of the utilization of food, of the kineticsof growth and metabolism in intact animals, and of the controllingmechanisms in the animal-food interaction.     

Key Aspects of Modern,Quantitative Drug Development

One of the main goals of modern drug development is customized care, where doctors match the right patient to the right treatment at the right dose, based on quantitative evidence. In this paper we review three key aspects of drug development that are critical towards achieving this goal. More specifically, we discuss (i) the advantages of modern model-based dose-finding as opposed to traditional pairwise comparisons, (ii) the value of pharmacometrical modeling, understanding the variability in how patients metabolize, tolerate, and respond to drugs, and (iii) the potential impact of enrichment strategies to identify study populations that are most likely to benefit from the investigational drug under development.     

Quantitative Aspects of the M Protein Capillary Precipitin Test

A capillary procedure for quantitatively determining M protein is described. Capillaries were filled with measured amounts of serum and streptococcal extract. The capillaries were incubated, and then centrifuged to pack the precipitates. The relative sizes of the precipates were compared by a determination of the weights of their paper images (obtained by reflection from a microscope). Meaningful dilution curves were determined by this method. Variations of pH from 6 to 8 had little effect on the M protein precipitin test, and the test was not seriously affected by variations of the NaCl concentration from 0.85 to 4.67%. The addition of divalent ions (Ca(++) and Mg(++)) did not influence the results. This test can be used to make quantitative comparisons of M protein preparations and to titrate type-specific antisera.     

Quantitative Measurement of GLUT4 Translocation to the Plasma Membrane by Flow Cytometry

Glucose is the main source of energy for the body, requiring constant regulation of its blood concentration. Insulin release by the pancreas induces glucose uptake by insulin-sensitive tissues, most notably the brain, skeletal muscle, and adipocytes. Patients suffering from type-2 diabetes and/or obesity often develop insulin resistance and are unable to control their glucose homeostasis. New insights into the mechanisms of insulin resistance may provide new treatment strategies for type-2 diabetes.The GLUT family of glucose transporters consists of thirteen members distributed on different tissues throughout the body¹. Glucose transporter type 4 (GLUT4) is the major transporter that mediates glucose uptake by insulin sensitive tissues, such as the skeletal muscle. Upon binding of insulin to its receptor, vesicles containing GLUT4 translocate from the cytoplasm to the plasma membrane, inducing glucose uptake. Reduced GLUT4 translocation is one of the causes of insulin resistance in type-2 diabetes^2,3.The translocation of GLUT4 from the cytoplasm to the plasma membrane can be visualized by immunocytochemistry, using fluorophore-conjugated GLUT4-specific antibodies.Here, we describe a technique to quantify total amounts of GLUT4 translocation to the plasma membrane of cells during a chosen duration, using flow cytometry. This protocol is rapid (less than 4 hours, including incubation with insulin) and allows the analysis of as few as 3,000 cells or as many as 1 million cells per condition in a single experiment. It relies on anti-GLUT4 antibodies directed to an external epitope of the transporter that bind to it as soon as it is exposed to the extracellular medium after translocation to the plasma membrane.     

A Quantitative Theory of Neural Computation

We show how a general quantitative theory of neural computation can be used to explain two recent experimental findings in neuroscience. The first of these findings is that in human medial temporal lobe there exist neurons that correspond to identifiable concepts, such as a particular actress. Further, even when such concepts are preselected by the experimenter, such neurons can be found with paradoxical ease, after examining relatively few neurons. We offer a quantitative computational explanation of this phenomenon, where apparently none existed before. Second, for the locust olfactory system estimates of the four parameters of neuron numbers, synapse numbers, synapse strengths, and the numbers of neurons that represent an odor are now available. We show here that these numbers are related as predicted by the general theory. More generally, we identify two useful regimes for neural computation with distinct ranges of these quantitative parameters.     

Quantitative Aspects of cGMP Phosphodiesterase Activation in Carp Rods and Cones

Cones are less light-sensitive than rods. We showed previously in carp that more light (>100-fold) is required in cones than in rods to activate 50% of cGMP phosphodiesterase (PDE). The lower effectiveness of PDE activation in carp cones is due partly to the fact that the activation rate of transducin (Tr) by light-activated visual pigment (R*) is 5-fold lower in carp cones than in rods. In this study, we tried to explain the remaining difference. First, we examined the efficiency of activation of PDE by activated Tr (Tr*). By activating PDE with known concentrations of the active (guanosine 5′-Ο-(γ-thio)triphosphate (GTPγS)-bound) form of Tr*, we found that Tr* activated PDE at a similar efficiency in rods and cones. Next, we examined the contribution of R* and Tr* lifetimes. In a comparison of PDE activation in the presence (with GTP) and absence (with GTPγS) of Tr* inactivation, PDE activation required more light (and was therefore less effective) when Tr* was inactivated in both rod and cone membranes. This is probably because inactivation of Tr* shortened its lifetime, thereby reducing the number of activated PDE molecules. The effect of Tr* inactivation was larger in cones, probably because the lifetime of Tr* is shorter in cones than in rods. The shorter lifetimes of Tr* and R* in cones seem to explain the remaining difference in the effectiveness of PDE activation between rods and cones.     

Quantitative Aspects of Antenna Grooming in Bees (Apoidea: Hymenoptera)

Antenna grooming in more than 100 species of bees (Apoidea), representing 34 genera of the 7 major bee families is recorded and quantitatively analysed. Most species of bees fall into one of two groups with respect to repetitive antenna cleaning: “Uniscrapers” predominantly clean their antennae with one stroke, “biscrapers” mostly with two subsequent strokes. Uniscrapers are more consequent in their behaviour than biscrapers. Most biscrapers occasionally clean their antenna with one or three strokes. Individual variation in the ratio of stroke repetition is considerably larger in bi- than in uniscrapers. In several species males and females differ with respect to their antenna cleaning behaviour, females tend to be more uniscraping. Most species of the families Colletidae, Halictidae, and Andrenidae, as well as the species of the genera Ceratina and Nomada (Anthophoridae) are biscrapers. Almost all species of Melittidae, Megachilidae, Apidae, and Anthophoridae (except Ceratina and Nomada) are uniscrapers. Bees with an antenna cleaner with ancestral (plesiomorphic) morphology are mostly, but not always, biscrapers, those with a derived antenna cleaner are always uniscrapers. Bees with a derived antenna cleaner perform on average less cleaning actions and strokes than those with an ancestral antenna cleaner. Uniscrapers with an ancestral strigilis do on average more cleaning actions per minute than biscrapers, thus they compensate partly for the fewer number of strokes. But nevertheless the uniscrapers do fewer strokes than the biscrapers (both with an ancestral strigilis). Females clean their antennae on average more often than males. It is interpreted that the behaviour of uniscraping and a derived morphology of the antenna cleaner result in greater efficiency than the status which is ancestral for Apoidea (biscraping and a plesiomorphic antenna cleaner).     

Towards a Quantitative Theory of Epidermal Calcium Profile Formation in Unwounded Skin

We propose and mathematically examine a theory of calcium profile formation in unwounded mammalian epidermis based on: changes in keratinocyte proliferation, fluid and calcium exchange with the extracellular fluid during these cells’ passage through the epidermal sublayers, and the barrier functions of both the stratum corneum and tight junctions localised in the stratum granulosum. Using this theory, we develop a mathematical model that predicts epidermal sublayer transit times, partitioning of the epidermal calcium gradient between intracellular and extracellular domains, and the permeability of the tight junction barrier to calcium ions. Comparison of our model’s predictions of epidermal transit times with experimental data indicates that keratinocytes lose at least 87% of their volume during their disintegration to become corneocytes. Intracellular calcium is suggested as the main contributor to the epidermal calcium gradient, with its distribution actively regulated by a phenotypic switch in calcium exchange between keratinocytes and extracellular fluid present at the boundary between the stratum spinosum and the stratum granulosum. Formation of the extracellular calcium distribution, which rises in concentration through the stratum granulosum towards the skin surface, is attributed to a tight junction barrier in this sublayer possessing permeability to calcium ions that is less than 15 nm s⁻¹ in human epidermis and less than 37 nm s⁻¹ in murine epidermis. Future experimental work may refine the presented theory and reduce the mathematical uncertainty present in the model predictions.     

Multiple Aspects of ATP-Dependent Nucleosome Translocation by RSC and Mi-2 Are Directed by the Underlying DNA Sequence

Background

Chromosome structure, DNA metabolic processes and cell type identity can all be affected by changing the positions of nucleosomes along chromosomal DNA, a reaction that is catalysed by SNF2-type ATP-driven chromatin remodelers. Recently it was suggested that in vivo, more than 50% of the nucleosome positions can be predicted simply by DNA sequence, especially within promoter regions. This seemingly contrasts with remodeler induced nucleosome mobility. The ability of remodeling enzymes to mobilise nucleosomes over short DNA distances is well documented. However, the nucleosome translocation processivity along DNA remains elusive. Furthermore, it is unknown what determines the initial direction of movement and how new nucleosome positions are adopted.

Methodology/Principal Findings

We have used AFM imaging and high resolution PAGE of mononucleosomes on 600 and 2500 bp DNA molecules to analyze ATP-dependent nucleosome repositioning by native and recombinant SNF2-type enzymes. We report that the underlying DNA sequence can control the initial direction of translocation, translocation distance, as well as the new positions adopted by nucleosomes upon enzymatic mobilization. Within a strong nucleosomal positioning sequence both recombinant Drosophila Mi-2 (CHD-type) and native RSC from yeast (SWI/SNF-type) repositioned the nucleosome at 10 bp intervals, which are intrinsic to the positioning sequence. Furthermore, RSC-catalyzed nucleosome translocation was noticeably more efficient when beyond the influence of this sequence. Interestingly, under limiting ATP conditions RSC preferred to position the nucleosome with 20 bp intervals within the positioning sequence, suggesting that native RSC preferentially translocates nucleosomes with 15 to 25 bp DNA steps.

Conclusions/Significance

Nucleosome repositioning thus appears to be influenced by both remodeler intrinsic and DNA sequence specific properties that interplay to define ATPase-catalyzed repositioning. Here we propose a successive three-step framework consisting of initiation, translocation and release steps to describe SNF2-type enzyme mediated nucleosome translocation along DNA. This conceptual framework helps resolve the apparent paradox between the high abundance of ATP-dependent remodelers per nucleus and the relative success of sequence-based predictions of nucleosome positioning in vivo.     

Cholesterol Translocation in a Phospholipid Membrane

Cholesterol (CHOL) molecules play a key role in modulating the rigidity of cell membranes and controlling intracellular transport and signal transduction. Using an all-atom molecular dynamics approach, we study the process of CHOL interleaflet transport (flip-flop) in a dipalmitoylphosphatidycholine (DPPC)-CHOL bilayer over a time period of 15 μs. We investigate the effect of the flip-flop process on mechanical stress across the bilayer and the role of CHOL in inducing molecular order in bilayer leaflets. The simulations are carried out at physiologically relevant CHOL concentration (30%), temperature (323 K), and pressure (1 bar). CHOL flip-flop events are observed with a rate constant of 3 × 10⁴ s⁻¹. Once a flip-flop event is triggered, a CHOL molecule takes an average of 73 nanoseconds to migrate from one bilayer leaflet to the other.     

γ-微管蛋白在猪卵母细胞成熟和活化中的分布(英)

微管蛋白（tubulin）是一蛋白质超家族,其中α-,β-微管蛋白是主要的微管蛋白,而γ-微管蛋白主要在微管组装中起作用. 我们利用蛋白质印迹和激光共聚焦技术研究了γ-微管蛋白在猪卵母细胞成熟、受精和活化中的分布. γ-微管蛋白存在于猪卵母细胞中,并且在减数分裂成熟各个时期的量保持不变. 它聚集在微管上,特别是中期纺锤体的两极和后末期的中板. 体外受精和孤雌活化后,γ-微管蛋白聚集在雌雄原核的周围.另外它也存在于精子的顶体帽和颈部.在早期卵裂中,γ-微管蛋白聚集在胚胎的细胞核周围.实验结果表明,γ-微管蛋白在猪卵母细胞、精子和胚胎的微管组装中起重要的调节作用,在猪受精过程中,精子和卵子都向受精卵贡献中心体物质.     

Quantitative Aspects of the 31P-NMR Detection of Manganese in Plant Tissues12

The uptake of manganese by maize roots was monitored in vivoby ³¹P nuclear magnetic resonance (NMR) spectroscopy and a quantitativeanalysis was developed on the basis of the line broadening ofthe vacuolar orthophosphate (P₁) signal. The line broadening,which was followed indirectly by measuring changes in the reciprocalpeak height of the P₁ signalin fully relaxed spectra, was foundto depend on pH and P₁ concentration, as well ason the presenceof organic acids, but for P₁ concentrations in the millimolarrange the method was sensitive to Mn₂₊ concentrations as lowas 0·1–1 µM. A linear relation was establishedbetween the reciprocal peak height of the vacuolar P₁ signalobserved in vivo and the total manganese content of the tissuedetermined subsequently by atomicabsorption. However, the paramagneticcontribution to the line widthobserved in vivo was much smallerthan expected from measurements on simple solutions and freeze-thawextracts and it was concluded that less than 5% of the manganesetaken up by the root tissue was present in the vacuoles as solubleMn²⁺. The ability to detect the free pool of divalent manganeseis one of several advantages of the ³¹P-NMR method relativeto the analogous¹H-NMR method based on the interaction betweenmanganese and water; and the non-invasive nature ofthe method,coupled with the potential to distinguish the cytoplasmic andvacuolar manganese fractions, allows the NMR method to complementthe information obtained by atomic absorption. Key words: Cytoplasm, intracellular compartmentation, manganese, ³¹P-NMR, vacuole     

Quantitative Aspects of Leaf Acid Phosphatase Activity and the Phosphorus Status of Tomato Plants

The relation between tomato leaf acid phosphatase activity andleaf tissue P content has been examined, and a study made ofthe effects of leaf development, variation in nitrogen supply,and variation in the growing medium on this relationship. Tomatoplants were grown in sand and given various concentrations ofphosphate. Plants were also grown for an initial period in peatcontaining an adequate level of phosphate, then transferredto peat to which was added 0 or 2.3 kg superphosphate m^–3and supplied with either 50 of 300 µg N ml^–1. Expressed on a unit tissue f. wt basis, acid phosphatase activityin the control plants in sand (given 41 µg P ml^minus;1)was highest in extracts from the expanding leaves and decreasedwith leaf maturity. However, when given a reduced supply ofphosphate, the enzyme activity in the more mature leaves wasequal to, or greater than, that in the expanding leaves. Thephosphatase activity increased first in the young, fully-expandedleaves and in the mature leaves (with 4.1 µg P ml^–1),but did not increase in the expanding leaves until the supplywas restricted to 2.1 µg P ml^–1. On closer examination,the increase in enzyme activity appeared to be associated withthe P level in the leaf tissues, the activity increasing whenthe level fell below about 0.25 per cent (g P per 100 g drywt tissue). The same relation was found with the plants grownin peat, and was independent of the concentration of nitrogensupplied to the plants. The fully expanded leaves showed the best enzyme response whenthe phosphate supply was restricted and the activity reflectedclosely the local levels of tissue P. The assay of the enzymein unpurified leaf extracts is simple and rapid, and could beused in a test to detect P-deficiency in tomato plants. Lycopersicon esculentum L, tomato, acid phosphatase activity, phosphorus status