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Lack of the metabotropic glutamate receptor subtype 7 selectively impairs short-term working memory but not long-term memory.

Metabotropic glutamate receptors (mGluRs), and in particular the mGluR group III receptors (subtypes 4, 6, 7, 8) are known to play a role in synaptic plasticity and learning. Here, we report the effect of mGluR7 gene ablation in different learning paradigms. In the acoustic startle response (ASR), no differences were seen between knockout (KO) mice and wildtype (WT) littermates in parameters including prepulse inhibition and habituation. In an open field test, no differences were seen between genotypes in motor activity, exploratory behaviour, and fearful behaviour. In a T-maze reinforced alternation working memory (WM) task, again no difference was seen between groups. However, when increasing the demands on working-memory in a 4-arm and 8-arm maze task, KO mice committed more WM errors than WT littermates thereby uncovering a highly significant difference between the two groups that persisted every day for the whole 9 days of the experiment. In a 4-arm maze with 2 arms baited, KO and wildtype mice committed the same number of LTM errors, whereas KOs committed more WM errors. Altogether, these findings suggest that a lack of mGluR7 mainly impairs short-term working but not long-term memory performance while having no effect on sensorimotor processing, non-associative learning, motor activity and spatial orientation. The effects on WM are task-dependent and become apparent in more complex but not simple learning tasks. We discuss how mGluR7 could influence WM.

Animals↗

Site-directed mutagenesis at the human B2 receptor and molecular modelling to define the pharmacophore of non-peptide bradykinin receptor antagonists.

Combining site-directed mutagenesis with information obtained from molecular modelling of the bradykinin (BK) human B2 receptor (hB2R) as derived from the bovine rhodopsin crystal structure [Science 289 (2000) 739], we previously defined a putative binding mode for the non-peptide B2 receptor antagonists, FR173657 and LF16-0687 [Can J Physiol Pharmacol 80 (2002) 303]. The present work is aimed to define the specific role of the quinoline moiety in the pharmacophore of these non-peptide antagonists. The effect of the mutations I110A, L114A (TM, transmembrane 3), W256A (TM6), F292A, Y295A and Y295F (TM7) was evaluated. None of the mutations affected the binding interaction of peptide ligands: the agonist BK and the peptide antagonist MEN 11270. The affinities in competing for [3H]-BK binding and in blocking the BK-induced IP production by the non-peptide antagonists LF16-0687 and FR173657 at the wild type and mutant receptors were analysed. While the affinities of LF16-0687 and FR173657 were crucially decreased at the I110A, Y295A, and Y295F mutants, the W256A mutation affected the affinity of the LF16-0687 only. The important contribution of the quinoline moiety was shown by the inability of an analogue of LF16-0687, lacking this moiety, to affect BK binding at the wild type receptor. On the other hand, the benzamidine group did not interact with mutated residues, since LF16-0687 analogues without this group or with an oxidated benzamidine displayed pairwise loss of affinity on wild type and mutated receptors. Further differences between FR173657 and LF16-0687 were highlighted at the I110 and Y295 mutants when comparing binding (pK(i)) and functional antagonist (pKB) affinity. First, the I110A mutation similarly impaired their binding affinity (250-fold), but at a less extent the antagonist potency of FR173657 only. Second, both the hydroxyl and the phenyl moieties of the Y295 residue had a specific role in the LF16-0687 interaction with the receptor, as demonstrated at the Y295F and Y295A mutants, respectively, but not in that of FR173657. Present data identify a receptor binding pocket comprised among TM3, 6, and 7, which concerns the interaction of the non-peptide antagonists FR173657 and LF16-0687, but not that of the peptide agonist or antagonist. Results indicate the quinoline group as the involved pharmacophoric element, and that the studied residues are differently involved in the interaction. The analysis performed by means of the GRID software led us to propose different spatial orientations of the quinoline moieties and partially overlapping binding pockets for the two ligands: that of LF16-0687 is located in the lipophilic environment amongst I110 (TM3), W256 (TM6), and Y295 (TM7) residues, whereas that of FR173657 lies essentially between I110 and Y295.

Animals↗

Alteration of human neuroblastoma cell morphology and neurite extension with micropatterns.

The spatial orientation of nerve cells plays a pivotal role in nerve regeneration. Here we report a new method for regulating neuronal cell morphology and guiding neurite extension on standard tissue culture dishes. Random copolymers of oligoethyleneglycol methacrylate and methacrylic acid [poly(OEGMA-co-MA)], microcontact printed on standard tissue culture dishes, resist cell attachment and remain intact in serum-containing medium for up to 2 weeks. Cell viability assay of SH-SY5Y cells demonstrated that poly(OEGMA-co-MA) on the substrate or in solution has no cytotoxic effect. When retinoic acid was added to SH-SY5Y cells, they extended neurites along the line patterns that are significantly longer than cells cultured on non-patterned culture dishes. The ability to guide neurite extension with micrometer precision is valuable for guiding directional growth of neurites and path finding of regenerating nerves.

Actins↗

Thrombomodulin induction in cultured human endothelial cells by 9-cis-locked retinoic acid analogues.

9-cis-Retinoic acid (RA) analogues devised to lock the 9-cis double bond by ring formation were synthesized using two stereoselective carbon-carbon bond formation reactions as key steps. The palladium-mediated Suzuki reaction was adopted to construct a 7E-double bond (RA numbering) and the Horner-Emmons olefination was employed for stereoselective 11E-double bond (RA numbering) formation. The synthesized 9-cis-RA analogues that are locked by five-membered ring systems (cyclopentene, dihydrofuran, and dihydrothiophene) were shown to have comparable thrombomodulin induction activities to that of 9-cis RA. Conformational analysis of these compounds showed their similarity to 9-cis RA in the spatial orientation of the side chain and the terminal carboxy group.

Alitretinoin↗

Sialyl Lewis(x) analogs based on a quinic acid scaffold as the fucose mimic.

(-)-Quinic acid was used as a starting material for the preparation of sialyl Lewis(x) mimetics in order to target E-selectin. Spatial orientation of the hydroxyl groups of quinic acid could mimic the l-fucose ones. Introduction of a side chain ending with a carboxylic acid was effected to replace the sialic acid interaction at the carbohydrate recognition domain. A first series of derivatives, incorporating amino acids linked to quinic acid, were tested for their affinity and found to interact with E-selectin with IC(50) within the millimolar range.

Cell Adhesion↗

Microvascular invasion during endochondral ossification in experimental fractures in rats.

In this study morphologic techniques have been used to detail the angiogenic response that accompanies endochondral fracture healing in a clinically relevant, reproducible rat model. In this displaced fracture, the gap fills with cartilage that later is replaced by bone, via endochondral ossification. A transient periosteal circulation, followed by a permanent medullary circulation accompany this progression. From 2 to 6 weeks, vessels grow out from the periosteal tissue and give rise to vascular buds, which abut directly onto the avascular zone corresponding to the fracture defect. From 3 weeks onwards, a second wave of vessels grows out from the marrow to the cartilage-filled fracture defect, terminating as vascular buds and loops lined by endothelial and perivascular cells. The loops and buds stain strongly for laminin but transmission electron microscopy does not demonstrate an identifiable basement membrane, pointing to a region of active extracellular matrix turnover. These vessels are intimately associated with osteoblasts and newly formed woven bone forming finger-like composite structures that protrude into the mineralized cartilage matrix with which they form a clearly demarcated interface. Invading vessels and woven bone successively replace the cartilage matrix to mediate repair. Both the vascular structures and progression of endochondral ossification observed, closely resemble those described in the normal epiphyseal growth plate, indicating that the fundamental processes are similar. However, there is a difference in the spatial orientation of cells such that the healing front in the fracture model is relatively disorganized, compared to the orderly linear array of cells at the epiphyseal growth plate.

Animals↗

Assembly of the otoconia complex to the macular sensory epithelium of the vestibule.

In the inner ear, specificity of stimulus perception is achieved by associating the sensory epithelia of the three mechanoreceptor organs, the utricle/saccule, cristae, and cochlea, with distinct types of acellular matrices. Only the utricle and saccule have an extremely dense matrix, the otoconial complex, which overlies the sensory epithelium (macula) and provides inertial mass to generate shearing forces essential for the mechanoreceptors to sense gravity and linear acceleration. Such sensation is necessary for spatial orientation and balance. The importance of otoconia is clearly demonstrated by the impact of balance disorders upon the elderly population that involve otoconia degeneration, as well as by canalithiasis and cupulolithiasis, in which otoconia are dislocated. This underscores the need to understand how otoconia are formed and maintained and how to prevent their degeneration. To date, a number of otoconia-related proteins have been identified mostly in mice and bony fish. Although most of these proteins are also present in other structures of the inner ear, a distinct collection of proteins in the macula plus the unique ionic microenvironment of the endolymph near its epithelium likely contribute to the site-specific calcification of otoconia. Based on the current literature and ongoing research, this mini-review postulates a working model of how the otoconia complex is assembled specifically above the macular sensory epithelium of the vestibule. The central hypothesis of this model is that proteins are critical in sequestering calcium for crystallization in the calcium-poor endolymph. The review also sets forth some issues that need to be resolved in the future.

Animals↗

Electron-conducting redox hydrogels: Design, characteristics and synthesis.

Redox hydrogels constitute the only electron-conducting phase in which water-soluble chemicals and biochemicals dissolve and diffuse. The combination of solubility and diffusion makes the electron-conducting gels permeable to water-soluble biochemicals and chemicals. The electron-conducting redox hydrogels serve to electrically connect the redox centers of enzymes to electrodes, enabling their use whenever leaching of electron-shuttling diffusional redox mediators must be avoided, which is the case in subcutaneously implanted biosensors for diabetes management and in miniature, potentially implantable, glucose-O2 biofuel cells. Because the hydrogels envelope the redox enzymes, they electrically wire the reaction centers to electrodes irrespective of spatial orientation and connect to electrode redox centers of multiple enzyme layers. Hence, the attained current densities of enzyme substrate electrooxidation or electroreduction are much higher than with enzyme monolayers packed onto electrode surfaces.

Electrons↗

MALDI-MS-based imaging of small molecules and proteins in tissues.

The direct analysis of tissues using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) enables both endogenous and exogenous compounds present in tissues to be detected with molecular specificity while maintaining their spatial orientation. This unique combination, coupled with excellent sensitivity and rapid analysis time, presents many potential advantages to a wide range of applications in diverse biological fields. Recent advances have shown how the technique can be applied to cancer research, neuroscience and pharmaceutical development. Examples include the use of unique protein profiles to classify human tumor tissues and predict patient outcomes, the discovery of protein changes in mouse cerebellum as a function of development, and the two-dimensional visualization of the distribution of a drug and first-pass metabolites in rat whole-body sections.

Animals↗

Progress in plasmodial differentiation improves regularity of oscillating contractions in Physarum polycephalum.

Based on the knowledge about subcellular morphogenetic processes in the acellular slime mold Physarum polycephalum, we hypothesized that during differentiation of undifferentiated endoplasm to the highly differentiated complex structure of the contractile apparatus of this organism, the regularity of oscillating contractions must improve. We measured the endogenous contraction automaticity starting from the de novo generation within minutes after sampling small portions of undifferentiated endoplasm. The standard deviation of the normalized period duration of these samples was compared to the respective values of radial contractions of differentiated protoplasmic plasmodial strands. The mean normalized standard deviation in endoplasmic drops was 28.3+/-12.2%. Respective values in protoplasmic strands were 10.0+/-3.7%. The difference between the experimental groups was highly significant (p<<0.0001). We interpret the verification of our hypothesis as an indication that the very regular oscillating contractions in fully differentiated stages of Physarum require the complex structure of the sophisticated contractile apparatus, represented by the circular plasmalemma invagination system of protoplasmic strands, while the regularity is lower in stages, where the differentiation is still in progress. We believe that this is due to deficits in coordination capabilities, which need a directional and spatially oriented protoplasmic streaming as a precondition.

Animals↗

Ontogenetic development of the helical heart: concepts and facts.

The structural and functional organization of the ventricular myocardial mass is a controversial matter that cannot be resolved by anatomical studies alone. Therefore, other approaches such as investigations of the ontogenetic development of the ventricular myocardium might help to resolve controversies about its structural and functional organization. It has recently been proposed that the spatial orientation of Torrent-Guasp's ventricular myocardial band model (basal and apical loops) might be the mature morphological correlate of twists and torsions of the embryonic heart loop. In the present contribution, the suggestions made in this concept were analyzed in the light of currently known facts about the development of the embryonic heart. It was found that some of the suggestions made in this concept do not correspond to embryological facts, whereas other suggestions could neither be disproved nor confirmed on the basis of our current knowledge on heart development. The answer to the question as to which of the various models of myocardial fibre organization fits best with the ontogenesis of the myocardial mass awaits future studies. The myocardial units of Torrent-Guasp's myocardial band model are said to have a functional rather than a morphological personality. Future studies on the ontogenetic development of the myocardium, therefore, should comprise not only anatomical analyses of dead specimens but should additionally comprise high resolution in vivo analyses of the development of the spatio-temporal contraction patterns of embryonic and fetal hearts.

Animals↗

A K52Q substitution in the globular domain of histone H1t modulates its nucleosome binding properties.

A comparison of the globular domain sequences of the somatic H1d and testis-specific H1t revealed a single substitution of lysine 52 in H1d to glutamine 54 in H1t, which is one of the three crucial residues within the second DNA binding site. The globular domains of both histones were modeled using the crystal structure of chicken GH5 as a template and was also docked onto the nucleosome structure. The glutamine residue in histone H1t forms a hydrogen bond with main chain carbonyl of methionine-52 (in H1t) and is spatially oriented away from the nucleosome dyad axis. A consequence of this change was a lower affinity of recombinant histone H1t towards Four-way junction DNA and reconstituted 5S mononucleosomes. When Gln-54 in Histone H1t was mutated to lysine, its binding affinity towards DNA substrates was comparable to that of histone H1d. The differential binding of histones H1d and H1t towards reconstituted mononucleosomes was also reflected in the chromatosome-stop assay.

Amino Acid Sequence↗

Comparative ultrastructure of collagen fibrils in uterine leiomyomas and normal myometrium.

OBJECTIVE: To examine the ultrastructural characteristics of extracellular matrix and mature collagen fibrils in uterine leiomyomas and compare them with those in adjacent normal myometrium. DESIGN: Analysis of paired leiomyoma-myometrium in surgical specimens. SETTING: Research center and tertiary care center. SUBJECT(S): Women undergoing medically indicated hysterectomy for symptomatic uterine leiomyomas. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Appearance and spatial orientation of the collagen fibrils in leiomyomas compared with myometrium. RESULT(S): Observation of specimens at x 12,500 magnification indicated that collagen fibrils were more abundant, loosely packed, and arrayed in a nonparallel manner in leiomyomas compared with myometrium. Random areas were examined at x 6,500 to x 64,000 magnification and revealed collagen fibrils of equal diameter in both leiomyomas and myometrium. However, an ordered and regular barbed appearance was present in collagen fibrils from myometrium but was lacking in leiomyomas. CONCLUSION(S): Leiomyomas contain an abnormal collagen fibril structure and orientation, which suggests that the well-regulated fibril formation in myometrium is altered in leiomyomas. Alterations in collagen genes may play a role in the pathogenesis of leiomyomas.

Adult↗

Assessment of information processing in children with functional articulation disorders.

OBJECTIVE: The aim of this study was to investigate the information processing skills in children who have functional articulation disorder by using a cognitive-linguistic test battery. MATERIAL-METHODS: Thirty-three children with functional articulation disorder were enrolled in this study. They ranged in age from 6 to 10 years. A control group without articulation disorders was composed of 160 children with similar properties. In the first stage of this study, both groups were administered the Ross Information Processing Assessment Test-Primary. In this test, the scores obtained from eight subtests; namely immediate memory, recent memory, recall of general information, spatial orientation, temporal orientation, organization, problem solving and abstract reasoning, was calculated for both groups. The scores of the eight subtests are combined to form the four composites. These composites are memory quotient, orientation quotient, thinking and reasoning quotient and information processing quotient. Information processing quotient is the best and the most comprehensive estimate of a child's overall information processing ability. In the second stage of the study the articulation-disordered group was divided into two subgroups according the number of the mistakes, which was done at the level of the phoneme. The scores obtained from both subgroups were compared with the scores of the control group. RESULTS: The overall statistical analysis of the scores revealed that; the study group had significantly lower scores than the control group from memory quotient, thinking and reasoning quotient and information processing quotient. When compared to control group, the subgroup who cannot pronounce one phoneme had similar scores from all four composites. Moreover, the children who cannot pronounce multiple phonemes had significantly lower scores from memory quotient, thinking and reasoning quotient and information processing quotient. The results obtained from this study seem to be suggesting that information processing skills of children with functional articulation disorder are significantly low as compared to normal children. CONCLUSIONS: These results are revealing that the information processing skills of children with functional articulation disorder should be investigated in a detailed manner. According to the results obtained from this investigation these children should be put on deficit oriented education programs in addition to articulation therapy.

Articulation Disorders↗

The superantigen staphylococcal enterotoxin A (SEA) and monoclonal antibody L243 share a common epitope but differ in their ability to induce apoptosis via MHC-II.

Crosslinking of MHC class II (MHC-II) molecules by antibodies or by superantigens (SAg) induces a variety of functional responses in the antigen presenting cell. We were able to allocate K39 as the residue that is essential for binding of antibody L243 to the alpha chain of HLA-DR. K39 is also essential for binding of staphylococcal enterotoxin A (SEA). However, the functional responses of the two ligands differ considerably exemplified by the ability of L243 to induce apoptosis in monocytic cells and in B cells, whereas SEA is unable to activate the apoptosis pathway. Despite the differences in functional responses, both ligands induce cell aggregation in MonoMac-1 cells. The SEA molecule with its two different binding sites associates one MHC alpha chain with one beta chain as opposed to two alpha chains that are brought into close proximity by the two identical antigen binding sites of L243. We therefore conclude that the spatial orientation of dimerized MHC-II and their associated proteins is an important factor for the nature of the transduced signal and consequently the outcome of functional responses.

Animals↗

Structural and functional anatomy of the globular domain of complement protein C1q.

C1q is the first subcomponent of the classical pathway of the complement system and a major connecting link between innate and acquired immunity. As a versatile charge pattern recognition molecule, C1q is capable of engaging a broad range of ligands via its heterotrimeric globular domain (gC1q) which is composed of the C-terminal regions of its A (ghA), B (ghB) and C (ghC) chains. Recent studies using recombinant forms of ghA, ghB and ghC have suggested that the gC1q domain has a modular organization and each chain can have differential ligand specificity. The crystal structure of the gC1q, molecular modeling and protein engineering studies have combined to illustrate how modular organization, charge distribution and the spatial orientation of the heterotrimeric assembly offer versatility of ligand recognition to C1q. Although the biochemical and structural studies have provided novel insights into the structure-function relationships within the gC1q domain, they have also raised many unexpected issues for debate.

Animals↗

Transesophageal real-time three-dimensional echocardiography methods and initial in vitro and human in vivo studies.

OBJECTIVES: The purpose of this study was to develop a transesophageal probe that: 1) enables on-line representation of the spatial structures of the heart, and 2) enables navigation of medical instruments. BACKGROUND: Whereas transthoracic real-time 3-dimensional (3D) echocardiography could recently be implemented, there is still no corresponding transesophageal system. Transesophageal real-time 3D echocardiography would have great potential for numerous clinical applications, such as navigation of catheters. METHODS: The newly developed real-time 3D system is based on a transesophageal probe in which multiple transducers are arranged in an interlaced pattern on a rotating cylinder. This enables continuous recording of a large echo volume of 70 mm in length and a sector angle of 120 degrees . The presentation of the volume-reconstructed data is made with a time lag of <100 ms. The frame rate is up to 20 Hz. In addition to conventional imaging, the observer can obtain a stereoscopic image of the structures examined with red/blue goggles. RESULTS: It was shown in vitro on ventricle- and aorta-form agar models and in vivo that the system enables excellent visualization of the 3D structures. Shape, spatial orientation, and the navigation of various catheters (e.g., EPS-catheter, Swan-Ganz-catheter), stents, or atrial septal defect occluders could be recorded on-line and stereoscopically depicted. The size of the echo sector enables a wide field of view without changing the position of the probe. CONCLUSIONS: Transesophageal real-time 3D echocardiography can be technically realized with the system presented here. The in vitro and in vivo studies show particularly the potential for navigation in the heart and large vessels on the basis of stereoscopic images.

Aorta↗

Numerical analysis of maximal bat performance in baseball.

Metal baseball bats have been experimentally demonstrated to produce higher ball exit velocity (BEV) than wooden bats. In the United States, all bats are subject to BEV tests using hitting machines that rotate the bat in a horizontal plane. In this paper, a model of bat-ball impact was developed based on 3-D translational and rotational kinematics of a swing performed by high-level players. The model was designed to simulate the maximal performance of specific models of a wooden bat and a metal bat when swung by a player, and included material properties and kinematics specific to each bat. Impact dynamics were quantified using the finite element method (ANSYS/LSDYNA, version 6.1). Maximum BEV from both a metal (61.5 m/s) and a wooden (50.9 m/s) bat exceeded the 43.1 m/s threshold by which bats are certified as appropriate for commercial sale. The lower BEV from the wooden bat was attributed to a lower pre-impact bat linear velocity, and a more oblique impact that resulted in a greater proportion of BEV being lost to lateral and vertical motion. The results demonstrate the importance of factoring bat linear velocity and spatial orientation into tests of maximal bat performance, and have implications for the design of metal baseball bats.

Acceleration↗