How does infliximab work in rheumatoid arthritis?

Since the initial characterization of tumor necrosis factor alpha (TNFalpha), it has become clear that TNFalpha has diverse biologic activity. The realization that TNFalpha plays a role in rheumatoid arthritis (RA) has led to the development of anti-TNF agents for the treatment of RA. Infliximab, a chimeric monoclonal antibody that specifically, and with high affinity, binds to TNFalpha and neutralizes the cytokine, is currently approved for the treatment of RA and Crohn's disease, another immune-inflammatory disorder. In addition to establishing the safety and efficacy of infliximab, clinical research has also provided insights into the complex cellular and cytokine-dependent pathways involved in the pathophysiology of RA, including evidence that supports TNFalpha involvement in cytokine regulation, cell recruitment, angiogenesis, and tissue destruction.     

How distributed is visual category information in human occipito-temporal cortex? An fMRI study

We used fMRI to study the distribution of object category information in the ventral visual pathway. Extending the findings of, we find that categories of stimuli can be distinguished by the pattern of activation they elicit across this entire pathway, even when the stimuli within a category differ in viewpoint, exemplar, or image format. However, regions within the ventral visual pathway are neither interchangeable nor equipotential. Although the FFA and PPA permit excellent discrimination between preferred versus nonpreferred stimuli (e.g., faces-bottles and houses-bottles, respectively), we find that neither region alone permits accurate discrimination between pairs of nonpreferred stimuli (e.g., bottles-shoes). These findings indicate that the ventral visual pathway is not homogeneous, but contains some regions (including FFA and PPA) that are primarily involved in the analysis of a single class of stimulus.     

How does inactivation change timing of replication in the human X chromosome?

Summary The kinetics of replication of the inactive (late replicating) X chromosome (LRX) were studied in karyotypically normal lymphocytes and human amniotic fluid cells. Both cell types were successively pulse labeled with 1-h or 1/2-h thymidine pulses in an otherwise BrdU-substituted S phase after partial synchronization of the cultures at G₁/S. For the first time with this technique, the entire sequence of replication was analyzed for the LRX from the beginning to the end of the S phase, with special reference to mid S (R-band to G-band transition replication). The inactive X is the last chromosome of the metaphase to start replication, with a delay of 1 or 2h, after which time a thymidine pulse results in R-type patterns. In mid S, the inactive X is the first chromosome to switch to G-type replication (without overlapping of both types and without any detectable replication pause). Until the end of S, a thymidine pulse results in G-type patterns. To rule out artifacts that might arise by the synchronization of cultures in these experiments, controls were carried out with BrdU pulses and the BrdU antibody technique without synchronization. In the course of replication, no fundamental difference was seen between the two different cell types examined. In contrast to studies using continuos labeling, this study did not reveal an interindividual difference of replication kinetics in the LRXs of the seven individuals studied; thus it is concluded that the inactive X chromosome shows only one characteristic course of replication.     

Are human taste thresholds similar on the right and left sides of the tongue?

The human tongue is a relatively symmetrical anatomical structure and is generally assumed functionally equivalent on both sides. Experimental evaluation of this assumption is complicated by the fact that psychophysical measurements tend to vary considerably across testing sessions. To address functional laterality, we determined the detection thresholds of six right-handed and six left-handed subjects for Na saccharin, NaCl, citric acid and quinine HCl. Five pairs of interwoven, left and right unilateral thresholds were obtained for each taste stimulus in 12 subjects (n = 480 separate thresholds). In most cases mean sensitivity based on multiple measurements was found to be laterally symmetrical, however, we observed a few cases of lateral asymmetry of both general and compound-specific sensitivity. Threshold values were found to vary considerably across sessions, consistent with the test-retest variability previously reported for whole mouth thresholds. We conclude that taste threshold sensitivity is equivalent on the left and right anterior tongue for most individuals. Given the occasional exceptions to this rule, however, it is advisable to employ a counterbalanced design for any experimental or clinical testing protocol in which treatments are applied asymmetrically to the tongue.     

How metals directly affect the antioxidant status in the liver and kidney of Oreochromis niloticus? An in vitro study

BackgroundMetals can disturb the integrity of physiological and biochemical mechanisms in fish. Thus components of defense as an antioxidant system are significant biomarkers due to their vital role in coping with metal stress. The aim of the current study is to investigate the direct effects of Cd, Cu, and Zn sublethal exposures (in vitro) on the antioxidant system parameters in the liver and kidney of Nile tilapia.MethodsThe antioxidant enzyme activities and GSH levels were analyzed after in vitro sublethal metal (200 and 400 μg/L Cd, Cu, and Zn) treatments of Oreochromis niloticus liver and kidney supernatants.ResultsMetals even at lower levels caused significant changes in the levels of antioxidant system parameters due to concentration, metal, and tissue type. GSH metabolism parameters were more responsive to the metal effect. TBARS levels and GPX activity were mostly increased while CAT, SOD, rGSH, and GSH/GSSG levels decreased. The kidney was more affected than the liver in vitro conditions. Cu was more effective in the liver whereas it was Zn for the kidney. Cd caused negative correlations among the antioxidant enzymes. Significant correlations were found between enzymes and GSH levels upon Zn and Cu exposures.ConclusionsDirect metal effects may trigger different response trends due to their nature and tissue differences. The current data provide a knowledge about which antioxidant biomarkers can define better the oxidative stress caused by direct metal effect for further studies including in vivo experiments.     

Sperm storage in the vertebrate female reproductive tract: How does it work so well?

The capacity for sperm storage within the female reproductive tract occurs widely across all groups of vertebrate species and is exceptionally well developed in some reptiles (maximum duration, 7 yr) and fish (maximum duration, >1 yr). Amphibians (most salamanders and one species of frog; duration approximately 5 mo), all birds examined to date and some bats, have also evolved the ability to store spermatozoa in the female reproductive tract. Although there are many reports on both the occurrence of female sperm storage and its adaptive benefits, few studies have been directed toward explaining the mechanisms involved. Phylogenetic evidence suggests that the capacity for sperm storage has evolved independently within different taxonomic groups, and it is by no means clear whether these groups have established similar or different mechanisms or whether simple and common principles have been exploited during evolution. If the process has indeed developed by the invention of numerous different and species-specific mechanisms, it is surprising that none have yet been elucidated by technologists wishing to improve the long-term storage of fresh semen. On the other hand, if there is a simple and common solution to the problem, readily accessed by diverse groups of species, it is equally logical to suppose that the mechanism should be easily discovered in the laboratory. While recognizing that studies on wild species are usually neither practically or ethically easy to undertake, it is clear that there is a huge and largely unexplored field to be investigated.     

How does stress affect human being—a molecular dynamic simulation study on cortisol and its glucocorticoid receptor

Stress can be either positive or negative to human beings. Under stressful conditions, the mental and physical conditions of human can be affected. There exists certain relation between stress and illness. The cortisol and other glucocorticoids bind to the same receptor, which is called glucocorticoid receptor. Some evidences indicated that cortisol molecule binding to its glucocorticoid receptor was necessary for the stress response. Up to now, the structure–function relationships between cortisol molecule and its glucocorticoid receptor have not been deliberated from the atomic-level. In order to get a detailed understanding of the structure–function relationships between the cortisol molecule and glucocorticoids receptor, we have carried out molecular dynamic (MD) simulations on glucocorticoid receptor (Apo system) and cortisol with its glucocorticoid receptor complex (HCY system). On the basis of molecular dynamic simulations, a couple of key residues were identified, which were crucial for the binding of cortisol molecule. The results of binding free energy calculations are in good agreement with the experiment data. Our research gives clear insights from atomic-level into the structural–functional aspects of cortisol molecule and its glucocorticoid receptor, and also provides valuable information for the design of drug which can treat stress related illnesses.     

Symmetry breakage in the vertebrate embryo: When does it happen and how does it work?

Asymmetric development of the vertebrate embryo has fascinated embryologists for over a century. Much has been learned since the asymmetric Nodal signaling cascade in the left lateral plate mesoderm was detected, and began to be unraveled over the past decade or two. When and how symmetry is initially broken, however, has remained a matter of debate. Two essentially mutually exclusive models prevail. Cilia-driven leftward flow of extracellular fluids occurs in mammalian, fish and amphibian embryos. A great deal of experimental evidence indicates that this flow is indeed required for symmetry breaking. An alternative model has argued, however, that flow simply acts as an amplification step for early asymmetric cues generated by ion flux during the first cleavage divisions. In this review we critically evaluate the experimental basis of both models. Although a number of open questions persist, the available evidence is best compatible with flow-based symmetry breakage as the archetypical mode of symmetry breakage.     

How far does phospholipase C activity depend on the cell calcium concentration? A study in intact cells.

The dependence of phospholipase C activity on the cytosolic Ca2+ concentration ([Ca2+]i) was studied in intact liver cells treated with the Ca2+-mobilizing hormone vasopressin, or not so treated. Phospholipase C (PLC) activity was estimated from the formation of [3H]inositol trisphosphate (InsP3) and the degradation of [3H]phosphatidylinositol 4,5-bisphosphate (PtdInsP2). The [Ca2+]i of the cells was clamped from 29 to 1130 nM by quin2 loading. This wide concentration range was obtained by loading the hepatocytes with a high concentration of the Ca2+ indicator in low-Ca2+ medium or by using the Ca2+ ionophore ionomycin in medium containing Ca2+. In resting cells, in which [Ca2+]i was 193 nM, treatment with 0.1 microM-vasopressin which stimulates liver PLC maximally, tripled InsP3 content and raised [Ca2+]i to 2 microM within 15 s. Lowering [Ca2+]i partially decreased cell InsP3 content as well as the ability of vasopressin to stimulate InsP3 formation maximally. At 29 nM, the lowest Ca2+ concentration obtained in isolated liver cells, basal InsP3 content was 64% of that measured in control cells. Addition of vasopressin no longer affected [Ca2+]i, but significantly increased InsP3 by 200%, although less than in the controls (300%). The maintenance of the greater part of the PLC response at constant [Ca2+]i indicated that, in the liver, InsP3 formation does not result from an increase in [Ca2+]i. The effects of lowering [Ca2+]i were reversible. When low cell [Ca2+]i was restored to a normal value, resting InsP3 content and the ability of vasopressin to stimulate InsP3 formation maximally by 300% were also restored. Raising [Ca2+]i from 193 to 1130 nM had little effect on the InsP3 content or the vasopressin-mediated increase in InsP3. In agreement with the stimulation of PLC activity by vasopressin, cell [3H]PtdInsP2 and total PtdInsP2 were degraded by application of this hormone for 15 s. In contrast, when [Ca2+]i was lowered to 29 nM, basal [3H]PtdInsP2 and total PtdInsP2 were increased by about 30%, [3H]PtdInsP2 was further increased by vasopressin, but total PtdInsP2 was not changed. These results show that, in intact hepatocytes, PLC is little affected by [Ca2+]i concentrations above 193 nM, but is partially dependent on Ca2+ below that value. They suggest that, in addition to activating PLC activity, vasopressin might stimulate PtdInsP2 synthesis, presumably via phosphatidylinositol-phosphate kinase, and that this pathway might predominate in cells with low [Ca2+]i.     

How and why does the proteome respond to microgravity?

For medical and biotechnological reasons, it is important to study mammalian cells, animals, bacteria and plants exposed to simulated and real microgravity. It is necessary to detect the cellular changes that cause the medical problems often observed in astronauts, cosmonauts or animals returning from prolonged space missions. In order for in vitro tissue engineering under microgravity conditions to succeed, the features of the cell that change need to be known. In this article, we summarize current knowledge about the effects of microgravity on the proteome in different cell types. Many studies suggest that the effects of microgravity on major cell functions depend on the responding cell type. Here, we discuss and speculate how and why the proteome responds to microgravity, focusing on proteomic discoveries and their future potential.     

How does airway inflammation modulate asthmatic airway constriction? An antigen challenge study.

During the late-phase (LP) response to inhaled allergen, mediators from neutrophils and eosinophils are released within the airways, resembling what occurs during an asthma attack. We compared the distribution of obstruction and degree of reversibility that follows a deep inspiration (DI) during early-phase (EP) and LP responses in nine asthmatic subjects challenged with allergen. Heterogeneity of constriction was assayed by determining frequency dependence of dynamic lung resistance and elastance, airway caliber by tracking airway resistance during a DI, and airway inflammation by measuring inflammatory cells in induced sputum postchallenge. Despite a paucity of eosinophils in the sputum at baseline (<1% of nonsquamous cells), asthmatic subjects showed a substantial EP response with highly heterogeneous constriction and reduced capacity to maximally dilate airways. The LP was associated with substantial airway inflammation in all subjects. However, five subjects showed only mild LP constriction, whereas four showed more marked LP constriction characterized by heterogeneous constriction similar to EP. Bronchoconstriction during LP was fully alleviated by administration of a bronchodilator. These findings, together with the impaired bronchodilatory response during a DI, indicate a physiological abnormality in asthma at the smooth muscle level and indicate that airway inflammation in asthma is associated with a highly nonuniform pattern of constriction. These data support the hypothesis that variability in responsiveness among asthmatic subjects derives from intrinsic differences in smooth muscle response to inflammation.     

How does microgravity affect the muscular and kinematic synergies in a complex movement?

The planning and the execution of voluntary movement relies on sensorimotor transformations in which representations of the external environment are integrated into motor programs. We studied executions of Whole Body Pointing movements, in normal and in transient microgravity (parabolic flights) conditions. Three processes could lead to adaptation to the new environmental condition: a radical change of terrestrial synergies, their partial modification or preservation. By applying a multivariate analysis on kinematic and electromyographic (EMG) data and by comparing the 1g and 0g conditions, our findings hint the hypothesis the descending information from vestibular system may be directed to change the synergies' modulation. An analogous analysis was performed on the kinematics: the invariance of intersegmental coordination among the segments' elevation angles suggests that these kinematic waveforms are used as reference signals to determine the appropriate muscle synergies in a subordinate and flexible manner in order to adapt to the novel mechanical constraints.     

How does the cerebral cortex work? Learning, attention, and grouping by the laminar circuits of visual cortex

The organization of neocortex into layers is one of its most salient anatomical features. These layers include circuits that form functional columns in cortical maps. A major unsolved problem concerns how bottom-up, top-down, and horizontal interactions are organized within cortical layers to generate adaptive behaviors. This article models how these interactions help visual cortex to realize: (i) the binding process whereby cortex groups distributed data into coherent object representations; (ii) the attentional process whereby cortex selectively processes important events; and (iii) the developmental and learning processes whereby cortex shapes its circuits to match environmental constraints. New computational ideas about feedback systems suggest how neocortex develops and learns in a stable way, and why top-down attention requires converging bottom-up inputs to fully activate cortical cells, whereas perceptual groupings do not.     

Smith‐Lemli‐Opitz syndrome in trisomy 13: How does the mix work?

BACKGROUND: Trisomy 13 and Smith-Lemli-Opitz syndrome (SLOS) are both well-recognized multiple congenital anomaly/mental retardation syndromes. CASE: In this report we describe a male newborn with trisomy 13 who also has features of SLOS, such as 2/3 toe syndactyly and a shawl-like scrotum. Biochemical analysis was consistent with SLOS, and limited molecular analysis revealed 1 mutation in the DHCR7 gene. CONCLUSIONS: The challenges in establishing the diagnosis of SLOS in this patient are presented and the unique coexistence of the 2 major malformation syndromes is discussed. Given the overlapping phenotype of the 2 syndromes, our report should encourage further research on cholesterol biosynthesis in patients with trisomy 13.     

Why does iron abrogate the cytotoxic effect of S-nitrosothiols on human and animal cultured cells?

It was found that dinitrosyl iron complexes (DNIC) with thiol-containing ligands (cysteine or glutathione) of concentrations up to 1 mM produce no cytotoxic effect on cultured cells from human milk gland carcinoma (MCF-7). The cytotoxic action on MCF-7 cells was produced by S-nitrosocysteine: at a concentration of 1 mM, it induced the death of 50% cells. A more stable S-nitrosothiol, S-nitrosoglutathione, did not produce any cytotoxic effect at the same concentration. It is assumed that the negative action of nitrosocysteine is due to its rapid degradation, which results in the accumulation of large amounts of free NO molecules followed by their oxidation by superoxide ions to peroxynitrite, an efficient inhibitor of metabolic processes. These processes seem to be not characteristic of the more stable S-nitrosoglutathione. The cytotoxic effect of nitrosocysteine was completlly abrogated by the addition of 0.2 mM ferrous citrate complex to the medium. When S-nitrosoglutathione NO (0.5 mM) or S-nitrosoglutathione (0.5 mM) + Fe(2+)-citrate (0.2 mM) were added to the medium, protein-bound dinitrosyl iron complexes formed with the involvement of endogenous or exogenous iron were detected in cells. The amount of the complexes in the presence of exogenous iron increased four times, reaching the value of 1.6 nmole/5 x 10(6) cells. Therefore, it was proposed that the blockade of the cytotoxic action of S-nitrosoglutathione by iron complexes is due to Cys-NO transformation of S-nitrosocysteine into dinitrosyl iron complexes. The high stability of these complexes ensures only a gradual accumulation of nitric oxide in cells.     

How does averaging affect protein structure comparison on the ensemble level?

Recent algorithmic advances and continual increase in computational power have made it possible to simulate protein folding and dynamics on the level of ensembles. Furthermore, analyzing protein structure by using ensemble representation is intrinsic to certain experimental techniques, such as nuclear magnetic resonance. This creates a problem of how to compare an ensemble of molecules with a given reference structure. Recently, we used distance-based root-mean-square deviation (dRMS) to compare the native structure of a protein with its unfolded-state ensemble. We showed that for small, mostly alpha-helical proteins, the mean unfolded-state Calpha-Calpha distance matrix is significantly more nativelike than the Calpha-Calpha matrices corresponding to the individual members of the unfolded ensemble. Here, we give a mathematical derivation that shows that, for any ensemble of structures, the dRMS deviation between the ensemble-averaged distance matrix and any given reference distance matrix is always less than or equal to the average dRMS deviation of the individual members of the ensemble from the same reference matrix. This holds regardless of the nature of the reference structure or the structural ensemble in question. In other words, averaging of distance matrices can only increase their level of similarity to a given reference matrix, relative to the individual matrices comprising the ensemble. Furthermore, we show that the above inequality holds in the case of Cartesian coordinate-based root-mean-square deviation as well. We discuss this in the context of our proposal that the average structure of the unfolded ensemble of small helical proteins is close to the native structure, and demonstrate that this finding goes beyond the above mathematical fact.