Axis formation in half blastoderms of the chick: Stage at separation and the relative positions of fused halves influence axis development

The blastoderm of the avian embryo acts during the early stages of development as an integrative system programmed to form a single embryonic axis. Isolated parts of the blastoderm are known to each form an axis, owing to the system's properties. In the work reported here, the regulative capability of the right and left halves of chick blastoderms to form an embryonic axis was examined systematically at different stages. This revealed a progressive change in the developing blastoderm. After early separation, the axis in each half will form at some distance from the blastoderm's original midline, while with late separation the axis will form next to the original midline and may even lack one row of somites at the medial rim. Since development stops in culture after about 2 days, axis development after early separation ceases before somites are formed, whereas after late separation somites and brain vesicles can develop. In addition, an attempt was made to learn whether the two halves of blastoderm, when shifted along the midline and then reunited in staggered fashion, act as a single or two separate embryonic fields. When reunion of the right and left halves was achieved so that the posterior end of one half was adjoining the posterior area pellucida region of the other half, a single embryonic axis developed. When, on the other hand, the shift was larger so that the posterior end was fused to the central area pellucida of the other half, two separated embryonic axes developed.     

Laterally Asymmetric Development of Duck Syrinx: Inhibition of Growth and Chondrogenesis of the Right Syringeal Half by the Left Half

For determination of whether position-related tissue interaction is involved in the laterally asymmetric development of the male duck syrinx, a cartilaginous vocal organ, the syrinxes from 10-day embryos were cut into right and left halves and cultured organotypically. The lateral halves developed equally when cultured separately, but their developments were unequal when they were cultured together in the same culture medium: that is, the right half developed less than the left half in terms of increase in size and chondrogenesis. When the right half from a normal or estrogen-treated embryo was cultured with the left half from a normal embryo, it accumulated less sulfated proteoglycans than when it was cultured with a left half from an estrogen-treated embryo. These results suggest that a soluble inhibitor that is susceptible to estrogen in the left half of the male syrinx inhibits the intrinsic increase in size and chondrogenesis of the right half and plays an important role in the laterally asymmetric development of the male duck syrinx.     

Morphology and fertilizability of zona-free hamster eggs separated into halves and quarters by centrifugation

When mature hamster eggs were freed from their zonae pellucidae and centrifuged in a Percoll gradient, each egg was separated into a light half and a heavy half. Chromosomes remained in their original position during centrifugation, resulting in the production of light and heavy halves with and without chromosomes. When the eggs were treated with cytochalasin D (CD) and then centrifuged, the chromosomes moved to the centripetal pole and were extruded rapidly before each egg separated into halves or fragments. In the eggs without CD treatment, the density of cortical granules was reduced in the centripetal region of the egg. In those treated with CD, the density of the granules was reduced in both centripetal and centrifugal regions of the egg. Both light and heavy halves were fertilizable. There was, however, a notable difference between light and heavy halves. Most of the heavy halves supported development of sperm nuclei into pronuclei, whereas only few of light halves could do so, suggesting that most of light halves were lacking or deficient in materials necessary for the development of a sperm (male) pronucleus. When the light and heavy halves were centrifuged further, each separated into two quarters. The lightest quarter, which was almost totally devoid of organelles, was buoyant and very fragile. Spermatozoa could fuse with it, but the incidence of the fusion was low. In this quarter, the sperm nucleus could decondense, but could not develop into a pronucleus. This was in marked contrast with other three quarters in which sperm nuclei could develop into well-formed pronuclei.     

Functional interaction sites of OprM with MexAB in the Pseudomonas aeruginosa multidrug efflux pump

Subunit-swapping between Pseudomonas aeruginosa MexAB-OprM and MexEF-OprN efflux pumps has shown that OprM can interact with MexEF to produce a functional efflux pump, but that OprN cannot functionally interact with MexAB. Taking advantage of this subunit selectivity, we carried out experiments using chimeric proteins composed of OprM and OprN to determine which regions of OprM are necessary for functional interaction with MexAB. We constructed two types of chimeric proteins: one with the N-terminal half of OprM and the C-terminal half of OprN (OprMN), and the second with these halves reversed (OprNM). Introduction of either of the chimeric protein genes into a mutant expressing MexEF alone restored the functionality of the efflux pump. However, expression of OprMN or OprNM in the presence of MexAB did not restore the pump functionality, indicating that the both the N- and C-terminal halves of OprM are necessary for a functional interaction with MexAB.     

Effects of external phosphorus supply on internal phosphorus concentration and the initiation,growth and exudation of cluster roots in Hakea prostrata R.Br.

The response of internal phosphorus concentration, cluster-root initiation, and growth and carboxylate exudation to different external P supplies was investigated in Hakea prostrata R.Br. using a split-root design. After removal of most of the taproot, equal amounts of laterals were allowed to grow in two separate pots fastened together at the top, so that the separate root halves could be exposed to different conditions. Plants were grown for 10 weeks in this system; one root half was supplied with 1 M P while the other halves were supplied with 0, 1, 25 or 75 M P. Higher concentrations of P supplied to one root half significantly increased the P concentration of those roots and in the shoots. The P concentrations in root halves supplied with 1 M P were invariably low, regardless of the P concentration supplied to the other root half. Cluster root initiation was completely suppressed on root halves supplied with 25 or 75 M P, whereas it continued on the other halves supplied with 1 M P indicating that cluster-root initiation was regulated by local root P concentration. Cluster-root growth (dry mass increment) on root halves supplied with 1 M P was significantly reduced when the other half was either deprived of P or supplied with 25 or 75 M P. Cluster-root growth was favoured by a low shoot P status at a root P supply that was adequate for increased growth of roots and shoots without increased tissue P concentrations. The differences in cluster-root growth on root halves with the same P supply suggest that decreased cluster-root growth was systemically regulated. Carboxylate-exudation rates from cluster roots on root halves supplied with 1 M P were the same, whether the other root half was supplied with 1, 25 or 75 M P, but were approximately 30 times faster when the other half was deprived of P. Estimates of root P-uptake rates suggest a rather limited capacity for down-regulating P uptake when phosphate was readily available.     

Noncomplementarity in base sequences between the cohesive ends of coliphages 186 and lambda and the formation of interlocked rings between the two DNA's

The half molecules of 186 DNA have been isolated by the Hg(II)–Cs₂SO₄ density gradient centrifugal ion technique. The buoyant densities of the two halves in CsCI at 25°C. are 1.713 and 1.709 g./cm.³, corresponding to GC contents of 54% and 50%, respectively. Similarly, 5-bromouracil labeled λ DNA halves were separated. The isolation of the four DNA halves made it possible to test for homology in base sequences between the cohesive ends of λ and those of 186. There was no indication of any significant homology in base sequences between the cohesive ends of the two DNA's, as indicated by the absence of a band with intermediate buoyant density in CsCI when either half of 186 DNA was annealed with either half of 5-bromouracil labeled λ DNA and then centrifuged. The lack of cohesion between the two DNA's made it possible to demonstrate unequivocally the formation of interlocked rings (catenanes) between the two DNA's. The existence of a dimeric catenane is evidenced by the formation of a species of intermediate buoyant density when 5-bromouracil labeled λ DNA is cyclized in the presence of cyclic 186 DNA of a relatively high concentration. The molecular weight of one DNA relative to the other can be calculated from the position of the dimeric catenane in a density gradient by using the method of Baldwin. The result was in complete agreement with our previous measurements from the sedimentation coefficients and by electron microscopy. The probability of dimeric catenane formation when one DNA is cyclized in the presence of another DNA is discussed. The experimental results agree with the theoretical expectation.     

Functional interaction between the N- and C-terminal halves of human hexokinase II.

Mammalian hexokinases (HKs) I-III are composed of two highly homologous approximately 50-kDa halves. Studies of HKI indicate that the C-terminal half of the molecule is active and is sensitive to inhibition by glucose 6-phosphate (G6P), whereas the N-terminal half binds G6P but is devoid of catalytic activity. In contrast, both the N- and C-terminal halves of HKII (N-HKII and C-HKII, respectively) are catalytically active, and when expressed as discrete proteins both are inhibited by G6P. However, C-HKII has a significantly higher Ki for G6P (KiG6P) than N-HKII. We here address the question of whether the high KiG6P of the C-terminal half (C-half) of HKII is decreased by interaction with the N-terminal half (N-half) in the context of the intact enzyme. A chimeric protein consisting of the N-half of HKI and the C-half of HKII was prepared. Because the N-half of HKI is unable to phosphorylate glucose, the catalytic activity of this chimeric enzyme depends entirely on the C-HKII component. The KiG6P of this chimeric enzyme is similar to that of HKI and is significantly lower than that of C-HKII. When a conserved amino acid (Asp209) required for glucose binding is mutated in the N-half of this chimeric protein, a significantly higher KiG6P (similar to that of C-HKII) is observed. However, mutation of a second conserved amino acid (Ser155), also involved in catalysis but not required for glucose binding, does not increase the KiG6P of the chimeric enzyme. This resembles the behavior of HKII, in which a D209A mutation results in an increase in the KiG6P of the enzyme, whereas a S155A mutation does not. These results suggest an interaction in which glucose binding by the N-half causes the activity of the C-half to be regulated by significantly lower concentrations of G6P.     

A nonhuman primate's perception of object relations: experiments on cottontop tamarins, Saguinus oedipus

Objects in nature often have spatial and functional relationships with other objects. For example, fruit may be connected to tree branches, bushes sometimes function as landmarks for home, and stones are functionally associated with nuts when they are used to crack the nuts open. Although animals may use the spatial and functional relationships between specific objects, it is important for ethologists interested in cognitive mechanisms to ask whether animals understand the spatial relationship between objects in a more general and abstract way. In this experiment, we ask whether a small New World monkey, the cottontop tamarin, is capable of perceiving the abstract relational concept of ‘connectedness’. Using a traditional operant paradigm, subjects were required to press one button to images with connected objects, and another button to images with separated objects. In Phase 1 of the experiment, subjects received training with only one connected and one separated image. Following training, probe images were presented in which features such as colour, texture and shape were systematically manipulated to determine which features were more important in stimulus classification. Accuracies and reaction times of responses were recorded. On the basis of their performance, the tamarins appeared to recognize that changes in the colour or texture of two objects plays no functional role in determining whether such objects are connected or separated. In contrast, changes in the shape and distance between two objects does play an important functional role, and the tamarins appeared to be sensitive to such changes. In Phase 2, subjects received training with a larger set of images. After this training, classification accuracy remained significantly above chance with most probes, suggesting that the tamarins had acquired a more general connectedness concept. In Phase 3, novel images were presented. Classification accuracy was significantly above chance for many of the novel images. Overall, results suggest that tamarins perceive spatial relationships between stimuli to some extent and can classify images based in part on their spatial relationship. Other recent experiments have shown that cottontop tamarins respond to spatial relationships even better than in the current experiment when they have to act directly on the objects. The difference between these results suggests that tamarins perceive the spatial relationships between objects more readily in the context of an ecologically valid problem.     

Identification of sequences in human transferrin that bind to the bacterial receptor protein, transferrin-binding protein B

Alignment of amino-acid sequences from the N-terminal and C-terminal halves of transferrin-binding protein B revealed an underlying bilobed nature with several regions of identity. Based on this analysis, purified recombinant fusion proteins of maltose-binding protein (Mbp) with intact TbpB, its N-terminal half or C-terminal half from the human pathogens Neisseria meningitidis and Moraxella catarrhalis were produced. Solid-phase binding assays and affinity isolation assays demonstrated that the N-terminal and C-terminal halves of TbpB could bind independently to human transferrin (hTf). A solid-phase overlapping synthetic peptide library representing the amino-acid sequence of hTf was probed with soluble, labelled Mbp-TbpB fusions to localize TbpB-binding regions on hTf. An essentially identical series of peptides from domains within both lobes of hTf was recognized by intact TbpB from both organisms, demonstrating a conserved TbpB-hTf interaction. Both halves of TbpB from N. meningitidis bound the same series of peptides, which included peptides from equivalent regions on the two hTf lobes, indicating that TbpB interacts with each lobe of hTf in a similar manner. Mapping of the peptide-binding regions on a molecular model of hTf revealed a series of nearly adjacent surface regions that nearly encircled each lobe. Binding studies with chimeric hTf/bTf transferrins demonstrated that regions in the C-lobe of hTf were preferentially recognized by the N-terminal half of TbpB. Collectively, these results provide evidence that TbpB consists of two lobes, each with distinct yet homologous Tf-binding regions.     

Val133 and Cys137 in transmembrane segment 2 are close to Arg935 and Gly939 in transmembrane segment 11 of human P-glycoprotein

P-glycoprotein (P-gp; ABCB1) transports a wide variety of structurally diverse compounds out of the cell. The protein has two homologous halves joined by a linker region. Each half consists of a transmembrane (TM) domain with six TM segments and a nucleotide-binding domain. The drug substrate-binding pocket is at the interface between the TM segments in each half of the protein. Preliminary studies suggested that the arrangement of the two halves of P-gp shows rotational symmetry (i.e. "head-to-tail" arrangement). Here, we tested this model by determining whether the cytoplasmic ends of TM2 and TM3 in the N-terminal half are in close contact with TM11 in the C-terminal half. Mutants containing a pair of cysteines in TM2/TM11 or TM3/TM11 were subjected to oxidative cross-linking with copper phenanthroline. Two of the 110 TM2/TM11 mutants, V133C(TM2)/G939C(TM11) and C137C(TM2)/A935C (TM11), were cross-linked at 4 degrees C, when thermal motion is reduced. Cross-linking was specific since no cross-linked product was detected in the 100 double Cys TM3/TM11 mutants. Vanadate trapping of nucleotide or the presence of some drug substrates inhibited cross-linking of mutants V133C(TM2)/G939C(TM11) and C137C(TM2)/A935C(TM11). Cross-linking of TM2 and TM11 also blocked drug-stimulated ATPase activity. The close proximity of TM2/TM11 and TM5/TM8 (Loo, T. W., Bartlett, M. C., and Clarke, D. M. (2004) J. Biol. Chem. 279, 7692-7697) indicates that these regions between the two halves must enclose the drug-binding pocket at the cytoplasmic side of P-gp. They may form the "hinges" required for conformational changes during the transport cycle.     

Interhemispheric asymmetry in rats differentiating figures by size

Motor alimentary conditioned reflex was elaborated in Wistar line rats to presentation of various pairs of figures, those of larger area serving as positive signals. In tests on animals with intact brain and with unilateral hemispheric cortical inactivation, new pairs of figures were used besides basic stimuli. The level of analysis in testing experiment was lower than normal: by 5.6% at intact brain, by 16.4%--at right hemisphere inactivation, by 36.4%--at left hemisphere inactivation. Elimination of the left hemisphere produced prolongation of the conditioned reaction time, signs of frustration and aftereffect. In conditions of right hemisphere inactivation the level of discrimination of figures depended on their specific properties and the side of presentation. Conclusion is made on dominating role of the left hemisphere in analysis of abstract and concrete characteristics of visual stimuli, in control of temporal, spatial and emotional characteristics of reactions in situations when considerable abstraction and distinguishing of generalizing invariant sign "more-less" is necessary for elaboration of the conditioned reflex.     

鱼类卵子发育能力的进一步研究

作者中的二人[Tung,and Tung.(童第周与叶毓芬)1944;Tung,Chang,andTung.(童第周、张致一与叶毓芬)1945]曾以分割的方法,研究金鱼(Carassiusauratus)卵子的发育能力。他们将2至8细胞时期的金鱼卵子,在去膜后,用玻璃针,按第一次或第二次的分裂平面,将整个卵子分割为二半。每半含有半数分裂球,一半卵黄,和一半未流入分裂球的细胞质。分割后,每半都能继续发育,但产生下列6种不同的情形:(1)二半都发育为完整的,一半大小的胚胎;(2)一个为完整的胚胎,另一个为不完整的胚胎;(3)一个为完整的胚胎,另一人为囊胚;     

Function of the MexB efflux-transporter divided into two halves

The MexA-MexB-OprM efflux pump exports structurally and functionally diverse xenobiotics and confers multi-drug resistance on Pseudomonas aeruginosa cells. The MexB transporter traverses the inner membrane twelve times, bears two large periplasmic domains and has two homologous tandem repeats. To test whether two homologous halves of MexB function independently or interdependently, the protein was divided medially into two halves, each consisting of six amino- and carboxyl-proximal transmembrane segments. When two halves of MexB were coexpressed from independent open reading frames, the cells lacking chromosomal mexB exhibited restored antibiotic resistance at a level close to that in the cells producing a full-length MexB. In contrast, MexB protein containing either an amino- or carboxyl-half fragment failed to transport antibiotics. To test whether the amino- and carboxyl-proximal halves were present in a complex, we purified the histidine-tagged carboxyl-proximal half molecule using nickel-chelate chromatography from the cells that coexpressed two halves. The results showed that the nonhistidine-tagged amino-proximal half was co-purified with the carboxyl-proximal half, thereby indicating that the amino-proximal half fragment was tightly associated with the carboxyl-proximal half molecule. These findings suggest that the presence of both amino- and carboxyl-halves of MexB in a complex is essential for transport activity.     

Categorical Perception of Fear and Anger Expressions in Whole,Masked and Composite Faces

Human observers are remarkably proficient at recognizing expressions of emotions and at readily grouping them into distinct categories. When morphing one facial expression into another, the linear changes in low-level features are insufficient to describe the changes in perception, which instead follow an s-shaped function. Important questions are, whether there are single diagnostic regions in the face that drive categorical perception for certain parings of emotion expressions, and how information in those regions interacts when presented together. We report results from two experiments with morphed fear-anger expressions, where (a) half of the face was masked or (b) composite faces made up of different expressions were presented. When isolated upper and lower halves of faces were shown, the eyes were found to be almost as diagnostic as the whole face, with the response function showing a steep category boundary. In contrast, the mouth allowed for a substantially lesser amount of accuracy and responses followed a much flatter psychometric function. When a composite face consisting of mismatched upper and lower halves was used and observers were instructed to exclusively judge either the expression of mouth or eyes, the to-be-ignored part always influenced perception of the target region. In line with experiment 1, the eye region exerted a much stronger influence on mouth judgements than vice versa. Again, categorical perception was significantly more pronounced for upper halves of faces. The present study shows that identification of fear and anger in morphed faces relies heavily on information from the upper half of the face, most likely the eye region. Categorical perception is possible when only the upper face half is present, but compromised when only the lower part is shown. Moreover, observers tend to integrate all available features of a face, even when trying to focus on only one part.     

Gravitation and the Hemispheric Specialization of the Brain in Bilateral Symmetry Determination

We studied the influence of weightlessness on bilateral symmetry detection during prolonged space flight. Supposing that weightlessness may affect visual information processing by the right and left hemispheres in different ways, we studied this phenomenon with regard for the part of the visual field where to a stimulus was presented (the sight fixation center or the left/right half of this field). We used two types of stimuli, i.e., closed figures (polygons) and distributed figures formed by dots. There was a distinct difference between the central and noncentral presentation of stimuli under terrestrial conditions. When a stimulus was presented noncentrally (on the left or right), a manifest dominance of the horizontal axis was observed. However, there was no substantial difference while stimulating the left and right parts of the visual field. This contradicts the hypothesis on hemispheric specialization of the brain in symmetry detection. When stimuli were presented eccentrically, weightlessness did not notably influence information processing. When they were presented centrally, the predominance of the vertical axis in closed figures tended to weaken under the impact of weightlessness. However, this predominance strengthened when multicomponent figures were presented in space. The different influences of weightlessness on perceiving symmetry of stimuli of different types shows that it may be detected at various levels with different degrees of using nonvisual sensory information.     

The Structure of Recombinant Human Annexin VI in Crystals and Membrane-bound

The crystal structure of calcium-free recombinant human annexin VI was solved at a resolution of 3.2 Å by using the annexin I model for Patterson search and refined to anR-factor of 19.0%. The molecule consists of two similar halves closely resembling annexin I connected by an α-helical segment and arranged perpendicular to each other. The calcium and membrane binding sites assigned by structural homology are therefore not located in the same plane. Analysis of the membrane-bound form of annexin VI by electron microscopy shows the two halves of the molecule coplanar with the membrane, but oriented differently to the crystal structure and suggesting a flexible arrangement. Ion channel activity has been found for annexin VI and the half molecules by electrophysiological experiments.     

Close Correlation of the Periodicities of the Cleavage Cycle and Cytoplasmic Cycle in Early Newt Embryos

The correlation between autonomous cyclic activity of the cytoplasm (cytoplasmic cycle) and the cleavage cycle was studied by using animal and vegetal half fragments of newt eggs formed by bisecting the uncleaved eggs after those eggs had been rotated through 90° off the vertical axis so as to alter the allocations of the cytoplasmic content in the two halves. When the bisection was made shortly after the rotation (Early Bisection), the resulting vegetal fragment showed 1.5 times longer intervals in the cytoplasmic cycle than its animal partner when cleavage was prevented by injection of colchicine, while when the bisection was made 30 min after the rotation (Late Bisection), the resulting pair of halves displayed equal intervals in the cytoplasmic cycle. The intervals of cell division of vegetal halves formed by the two kinds of bisection in the absence of colchicine were then examined. In these conditions, the vegetal half formed by Early Bisection still showed 1.5 times longer intervals in the cleavage cycle than its counterpart, and the half formed by Late Bisection displayed the same intervals in the cleavage cycle as its animal partner.     

The asymmetry of the free amino acid pool in sections of the rat brain

The content of free amino acid pool in symmetric regions of cortex, hypothalamus, midbrain and blood of rats which had produced the movement conditioned reflex strengthened by feeding was studied. It was established that the "untrained" rats have higher content of free amino acid pool in their blood. The brain of the experimental animals revealed the biochemical asymmetry which was marked by the differences in free amino acid pool distribution between the left and right halves of the studied regions of brain. It was shown that left sided asymmetry animals i.e. animals with heightened content of free amino acid pool in the left half of the brain dominated in the "untrained" group of rats. The supposition was expressed that this biochemical asymmetry may contribute to the ability of animals to learn.