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PubMed2002

Möbius and Möbius-like patients: etiology, diagnosis, and treatment options.

The surgical goal in Mobius patients is far more modest and differs from patients with unilateral developmental facial paralysis. It is impossible to restore a true smile in these mask-like, expressionless faces. Despite sophisticated microneurovascular transplantations, movement can only be restored along one vector and enhanced firmness in the cheeks, thus multiple differentiated facial animation is not achievable. A detailed neurological evaluation can identify possible motor donors or residual function, which can be used for additional dynamic restorations. Due to the multiple cranial nerve involvement a thorough clinical and electrophysiological examination is mandatory. In addition, electromyographic survey of the potential motor donors is very helpful to avoid weak wasted regeneration and prevent further downgrading of function. Because of the variety of cranial nerves involved in Möbius' syndrome, a standard procedure for dynamic restoration cannot and should not be promoted; instead, a careful preoperative objective and quantitative assessment should guide the reconstructive surgeon to the optimal reconstruction strategy. Useful movement can be restored in afflicted patients that may signal physical and psychological rehabilitation.

Adult↗

PubMed2001

Critical role for the EB1 and APC interaction in the regulation of microtubule polymerization.

Human EB1 was originally cloned as a protein that interacts with the COOH terminus of adenomatous polyposis coli (APC). Interestingly, this interaction is often disrupted in colon cancer, due to mutations in APC. EB1 also interacts with the plus-ends of microtubules and targets APC to microtubule tips. Since APC is detected on the kinetochores of chromosomes, it has been hypothesized that the EB1-APC interaction connects microtubule spindles to the kinetochores and regulates microtubule stability. In yeast, EB1 regulates microtubule dynamics, and its binding domain in APC may be conserved in Kar9, an EB1 binding protein involved in the microtubule-capturing mechanism. These results suggest that the interaction of EB1 and APC is important and may be conserved. However, it is largely unknown whether the EB1-APC interaction affects microtubule dynamics. Here, we show that EB1 potently promotes microtubule polymerization in vitro and in permeabilized cells, but, surprisingly, only in the presence of the COOH-terminal EB1 binding domain of APC (C-APC). Significantly, this C-APC activity is abolished by phosphorylation, which also disrupts its ability to bind to EB1. Furthermore, yeast EB1 protein effectively substitutes for the human protein but also requires C-APC in promoting microtubule polymerization. Finally, C-APC is able to promote microtubule polymerization when stably expressed in APC mutant cells, demonstrating the ability of C-APC to promote microtubule assembly in vivo. Thus, the interaction between EB1 and APC plays an essential role in the regulation of microtubule polymerization, and a similar mechanism may be conserved in yeast.

3T3 Cells↗

PubMed2002

Actin dynamics: tropomyosin provides stability.

Tropomyosin stabilizes actin filaments, while ADF/cofilin promotes their disassembly. These two proteins compete with each other for binding to the actin filament, providing a critical balance for actin assembly in vivo.

Actin Cytoskeleton↗

PubMed1996

Polysialylation of NCAM by a single enzyme.

The addition of poly-alpha2,8-N-acetylneuraminic acid (polysialic acid; PSA) to the neural cell adhesion molecule NCAM plays a crucial role in neural development [1-3], neural regeneration [4], and plastic processes in the vertebrate brain associated with neurite outgrowth [5], axonal pathfinding [6], and learning and memory [7,-9]. PSA levels are decreased in people affected by schizophrenia [10], and PSA has been identified as a specific marker for some neuroendocrine and lymphoblastoid tumours [11-13]; expression of PSA on the surface of these tumour cells modulates their metastatic potential [11-13]. Studies aimed at understanding PSA biosynthesis and the dynamics of its production have largely been promoted by the cloning of polysialyltransferases (PST-1 in hamster; PST in human and mouse) [14-16]. However, the number of enzymes involved in the biosynthesis of PSA has not been identified. Using incompletely glycosylated NCAM variants and soluble recombinant glycosyltransferases, we reconstituted the site at which PST-1 acts to polysialylate NCAM in vitro. The data presented here clearly demonstrate that polysialylation of NCAM is catalyzed by a single enzyme, PST-1, and that terminal sialylation of the N-glycan core is sufficient to generate the PSA acceptor site. Our results also show that PST-1 can act on core structures with the terminal sialic acid connected to galactose via an alpha2,3 or alpha2,6 linkage.

Animals↗

PubMed1997

Roles of the methane monooxygenase reductase component in the regulation of catalysis.

The reductase component (MMOR) of the soluble methane monooxygenase isolated from Methylosinus trichosporium OB3b catalyzes transfer of 2e- from NADH to the hydroxylase component (MMOH) where oxygen activation and substrate oxidation occur. It is shown here that MMOR can also exert regulatory effects on catalysis by binding to MMOH or to the binary complex of MMOH and component B (MMOB), another regulatory protein. MMOR alters the oxidation-reduction potentials of the dinuclear iron cluster at the active site of MMOH. Although little change is observed in the potential for the first electron transfer to the cluster (E(1)0' = 76 mV), the E(2)0' potential value for the second electron transfer is increased from 21 to 125 mV. This shift provides a larger driving force for electron transfer from MMOR and favors transfer of two rather than one electron as required by catalysis. Similar positive shifts in potential are observed even in the presence of MMOB which has been shown to cause a 132 mV negative shift in the midpoint potential of MMOH in the absence of MMOR. MMOR is also shown to decrease the rate of reaction between the fully reduced MMOH-MMOB and O2 approximately 20-fold at 4 degrees C. However, the time course of the key catalytic cycle intermediate that can react with substates, compound Q, is unaffected. This implies a compensating faster decay of one or more of the intermediates that occur between diferrous MMOH and compound Q in the reaction cycle, thereby limiting potential nonproductive autodecay of these intermediates. Accordingly, an increase in single turnover product yield is observed in the presence of MMOR. Interestingly, MMOR can cause the redox potential increases, changes in rates, and the increase in product yield when present at only 10% of the concentration of MMOH active sites. Substrate binding is shown to induce negligible changes in the redox potentials. Two alternative regulatory schemes are presented based on (i) thermodynamic coupling of component binding and redox changes or (ii) dynamic interconversion of two states of MMOH promoted by MMOR.

Catalysis↗

PubMedDate not supplied

Postoperative rupture of an aortic aneurysmal dilation associated with a patent ductus arteriosus in a dog.

OBJECTIVE: To describe an unusual case of fatal aortic aneurysm rupture after standard ligation of a left-to-right shunting patent ductus arteriosus (PDA) and review the literature concerning the causes of aneurysm and applicable medical physics of blood flow that may contribute to mural failure. STUDY DESIGN: Clinical case report. SAMPLE POPULATION: An 11-month-old female Brittany spaniel. RESULTS: A PDA was diagnosed in the dog after referral for a grade IV/IV cardiac murmur. Diagnostic tests performed included a minimum database, thoracic radiography, and echocardiography. Conventional surgical ligation of the PDA was performed. The postoperative recovery from anesthesia was uneventful. Approximately 5 hours after surgery, the dog vocalized once and collapsed. All resuscitative efforts failed. Necropsy showed a large, spiral rupture of the cranial and dorsal wall of the aorta that extended from the left subclavian artery to 2 cm distal to the PDA-aorta junction. No evidence of intramural, perianeurysmal hemorrhage or inflammation was noted. Histopathologic examination of the ruptured aneurysm showed that the wall lacked a defined tunica intima and endothelial cell layer. A distinct separation of the thinned tunica media parallel to the elastic fibers was noted. CONCLUSIONS AND CLINICAL RELEVANCE: Dilatation of the aorta near the area of a PDA is one of the recognized abnormalities associated with the condition. Aneurysmal dilatation results from and contributes to abnormal blood flow dynamics and increased aortic wall tension, which promotes expansion. Although an unusual and previously unreported sequela, rupture of the aneurysm should be considered a possible complication when ligation of a PDA with a conspicuous aortic aneurysmal dilation is performed.

Animals↗

PubMed2003

[Health promotion--a new approach in psychiatry].

Since its beginnings, the ideology of psychiatry has been determined by the paradigm of a disease model. Practically all psychiatric schools are characterized by their orientation towards a morbid, atypical behaviour. This, however, inevitably narrows the view of the soul of a person. Despite mental illness, resources are always present, these though are not recognized and not utilized. The theory of salutogenesis with its search for health preserving factors has significantly expanded insight on dynamic mental processes. The resulting concept of health promotion attempts to develop patient's self-healing powers by identifying healthy parts and supporting them by instructing patients in active participation. From a health promotion viewpoint, it is necessary to intervene at three levels: 1. At the patient level in the form of guidance and help to actively participate in the healing process. 2. At the staff level in the form of creating healthy work conditions and the promotion of active participation in the creation of this work environment. 3. At the level of psychiatric institutions in the form of promoting efficient communications structures within and between the various services. This paper describes the concept of health promotion in psychiatric services and its possible influence on the reshaping of theory and practice in psychiatry--necessary in the face of current challenges.

Adaptation, Psychological↗

PubMed2006

Phosphorylation and cleavage of presenilin-associated rhomboid-like protein (PARL) promotes changes in mitochondrial morphology.

Remodeling of mitochondria is a dynamic process coordinated by fusion and fission of the inner and outer membranes of the organelle, mediated by a set of conserved proteins. In metazoans, the molecular mechanism behind mitochondrial morphology has been recruited to govern novel functions, such as development, calcium signaling, and apoptosis, which suggests that novel mechanisms should exist to regulate the conserved membrane fusion/fission machinery. Here we show that phosphorylation and cleavage of the vertebrate-specific Pbeta domain of the mammalian presenilin-associated rhomboid-like (PARL) protease can influence mitochondrial morphology. Phosphorylation of three residues embedded in this domain, Ser-65, Thr-69, and Ser-70, impair a cleavage at position Ser(77)-Ala(78) that is required to initiate PARL-induced mitochondrial fragmentation. Our findings reveal that PARL phosphorylation and cleavage impact mitochondrial dynamics, providing a blueprint to study the molecular evolution of mitochondrial morphology.

Amino Acid Sequence↗

PubMed1997

Hydrolysis of gamma:epsilon isopeptides by cytosolic transglutaminases and by coagulation factor XIIIa.

Nepsilon-(gamma-glutamyl)lysine cross-links, connecting various peptide chain segments, are frequently the major products in transglutaminase-catalyzed reactions. We have now investigated the effectiveness of these enzymes for hydrolyzing the gamma:epsilon linkage. Branched compounds were synthesized, in which the backbone on the gamma-side of the cross-bridge was labeled with a fluorophor (5-(dimethylamino)-1-naphthalenesulfonyl or 2-aminobenzoyl) attached through an epsilon-aminocaproyl linker in the N-terminal position, and the other branch of the bridge was constructed with Lys methylamide or diaminopentane blocked by 2,4-dinitrophenyl at the Nalpha position. Hydrolysis of the cross-link could be followed in these internally quenched substrates by an increase in fluorescence. In addition to the thrombin and Ca2+-activated human coagulation Factor XIIIa, cytosolic transglutaminases from human red cells and from guinea pig liver were tested. All three enzymes were found to display good isopeptidase activities, with Km values of 10(-4) to 10(-5) M. Inhibitors of transamidation were effective in blocking the hydrolysis by the enzymes, indicating that expression of isopeptidase activity did not require unusual protein conformations. We suggest that transglutaminases may play a dynamic role in biology not only by promoting the formation but also the breaking of Nepsilon-(gamma-glutamyl)lysine isopeptides.

Animals↗

PubMed2001

Cytoplasmic tail-dependent internalization of membrane-type 1 matrix metalloproteinase is important for its invasion-promoting activity.

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is an integral membrane proteinase that degrades the pericellular extracellular matrix (ECM) and is expressed in many migratory cells, including invasive cancer cells. MT1-MMP has been shown to localize at the migration edge and to promote cell migration; however, it is not clear how the enzyme is regulated during the migration process. Here, we report that MT1-MMP is internalized from the surface and that this event depends on the sequence of its cytoplasmic tail. Di-leucine (Leu571-572 and Leu578-579) and tyrosine573 residues are important for the internalization, and the mu2 subunit of adaptor protein 2, a component of clathrin-coated pits for membrane protein internalization, was found to bind to the LLY573 sequence. MT1-MMP was internalized predominantly at the adherent edge and was found to colocalize with clathrin-coated vesicles. The mutations that disturb internalization caused accumulation of the enzyme at the adherent edge, though the net proteolytic activity was not affected much. Interestingly, whereas expression of MT1-MMP enhances cell migration and invasion, the internalization-defective mutants failed to promote either activity. These data indicate that dynamic turnover of MT1-MMP at the migration edge by internalization is important for proper enzyme function during cell migration and invasion.

Animals↗

PubMed2002

Cytoplasmic linker proteins promote microtubule rescue in vivo.

The role of plus end-tracking proteins in regulating microtubule (MT) dynamics was investigated by expressing a dominant negative mutant that removed endogenous cytoplasmic linker proteins (CLIPs) from MT plus ends. In control CHO cells, MTs exhibited asymmetric behavior: MTs persistently grew toward the plasma membrane and displayed frequent fluctuations of length near the cell periphery. In the absence of CLIPs, the microtubule rescue frequency was reduced by sevenfold. MT behavior became symmetrical, consisting of persistent growth and persistent shortening. Removal of CLIPs also caused loss of p150Glued but not CLIP-associating protein (CLASP2) or EB1. This result raised the possibility that the change in dynamics was a result of the loss of either CLIPs or p150Glued. To distinguish between these possibilities, we performed rescue experiments. Normal MT dynamics were restored by expression of the CLIP-170 head domain, but p150Glued was not recruited back to MT plus ends. Expression of p150Glued head domain only partially restored MT dynamics. We conclude that the CLIP head domain is sufficient to alter MT dynamics either by itself serving as a rescue factor or indirectly by recruiting a rescue factor. By promoting a high rescue frequency, CLIPs provide a mechanism by which MT plus ends may be concentrated near the cell margin.

Animals↗

PubMed2005

Complementary medicine and genetic medicine: polar disciplines or dynamic partners?

For more than a decade, a powerful movement promoting the integration of complementary and alternative medicine (CAM) and conventional medicine has evolved. Throughout the same period, there has been a monumental shift in the biologic sciences, and in perspectives on disease, resulting from advances in genetics. It is noteworthy, and perhaps not coincidental, that these "movements" have been occurring in parallel. The simultaneous growth of complementary medicine and genetic medicine may be fueled by a deep interest in the development of "personalized" medicine. There is a prevailing view that the metaphysical visions of these two fields are in conflict. To advance discussion of this question, we describe what we believe are the common philosophies and goals of these apparently disparate fields, and why it would be advantageous for them to work together in the service of the public's health.

Complementary Therapies↗

PubMed2006

Effects of feed additives and mixed eimeria species infection on intestinal microbial ecology of broilers.

Evaluation of digestive microbial ecology is necessary to understand effects of growth-promoting feed. In the current study, the dynamics of intestinal microbial communities (MC) were examined in broilers fed diets supplemented with a combination of antibiotic (bacitracin methylene disalicylate) and ionophore (Coban 60), and diets containing 1 of 2 essential oil (EO) blends, Crina Poultry (CP) and Crina Alternate (CA). Five treatments were analyzed: 1) unmedicated uninfected control; 2) unmedicated infected control; 3) feed additives monensin (bacitracin methylene disalicylate) + monensin (Coban 60; AI); 4) EO blend CP; and 5) EO blend CA. Additives were mixed into a basal feed mixture, and EO were adjusted to 100 ppm. Chicks were infected by oral gavage at 19 d of age with Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Duodenal, ileal, and cecal samples were taken from 12 birds per treatment just before and 7 d after challenge; 2 samples each were pooled to give a final number of 6 samples total; and all pooled samples were frozen until used for DNA extraction. Denaturing gradient gel electrophoresis was used to examine PCR-amplified fragments of the bacterial 16S ribosomal DNA variable region. Results are presented as percentages of similarity coefficients (SC). Dendrograms of PCR amplicon or band patterns indicated MC differences due to intestinal location, feed additives, and cocci challenge. Essential oil blends CP and CA affected MC in all gut sections. Each EO had different effects over MC, and they differed in most instances from the AI group. The cocci challenge caused drastic MC population shifts in duodenal, ileal, and cecal sections (36.7, 55.4, and 36.2% SC, respectively). Diets supplemented with CP supported higher SC between pre- and postchallenge MC (89.9, 83.3, and 76.4%) than AI (81.8., 57.4, and 60.0%). We concluded that mixed coccidia challenge caused drastic shifts in MC. These EO blends modulated MC better than AI, avoiding drastic shifts after a mixed challenge.

Animal Feed↗

PubMed1995

Vascular endothelial cell activation and permeability responses to thrombin.

The serine protease, thrombin, evokes numerous endothelial cell responses which regulate hemostasis, thrombosis and vessel wall pathophysiology. One such response, the development of intercellular gap formation and vascular permeability is relevant to each of these processes and is a cardinal features of inflammation. Regulation of endothelial cell gap formation and therefore permeability is a function of a dynamic balance between competing adhesive, barrier-promoting tethering forces and contractile, tension-producing forces which result in barrier dysfunction. The key tethering events governing focal endothelial cell adhesion to the extracellular matrix and cell-cell interactions are poorly understood. In contrast, information is rapidly increasing regarding endothelial-specific contractile processes driven by the actomyosin molecular motor. The level of myosin light chain phosphorylation catalyzed by a unique myosin light chain kinase promotes productive actin-myosin interaction and governs the degree of centripetal tension produced. In this review the signal transducing and contractile mechanisms by which thrombin elicits endothelial cellular activation through its specific receptor are addressed. The pathways by which thrombin may alter the balance between contractile and tethering forces to promote endothelial cell gap formation are discussed.

Animals↗

PubMed2006

TRP channels in C. elegans.

The TRP (transient receptor potential) superfamily of cation channels is present in all eukaryotes, from yeast to mammals. Many TRP channels have been studied in the nematode Caenorhabditis elegans, revealing novel biological functions, regulatory modes, and mechanisms of localization. C. elegans TRPV channels function in olfaction, mechanosensation, osmosensation, and activity-dependent gene regulation. Their activity is regulated by G protein signaling and polyunsaturated fatty acids. C. elegans TRPPs related to human polycystic kidney disease genes are expressed in male-specific neurons. The KLP-6 kinesin directs TRPP channels to cilia, where they may interact with F0/F1 ATPases. A sperm-specific TRPC channel, TRP-3, is required for fertilization. Upon sperm activation, TRP-3 translocates from an intracellular compartment to the plasma membrane to allow store-operated Ca2+ entry. The TRPM channels GON-2 and GTL-2 regulate Mg2+ homeostasis and Mg2+ uptake by intestinal cells; GON-2 is also required for gonad development. The TRPML CUP-5 promotes normal lysosome biogenesis and prevents apoptosis. Dynamic, precise expression of TRP proteins generates a remarkable range of cellular functions.

Animals↗

PubMedDate not supplied

Developing critical care skills for nurses in the ward environment: a work-based learning approach.

An account of collaborative working between an NHS trust and university in responding to the critical care agenda. An 'Introduction to Critical Care Skills' course initiative, which addresses the needs of nurses caring for level 1 and 2 patients in ward areas, is discussed. Work-based learning forms the focus of skills development using core competencies related to a holistic approach to caring for patients with complex needs. A dynamic evolving process of course development is promoting quality care for patients and closely reflects the needs of those caring for acutely ill patients outside the designated critical care environment.

Clinical Competence↗

PubMed2005

Are health education meetings effective in recruiting women in cervical screening programmes? An innovative and inexpensive intervention from the island of Crete.

INTRODUCTION: Cervical screening programs have been introduced in many countries and are generally regarded as the most appropriate and effective method currently available for preventing cervical cancer. Although action has been undertaken by some rural practitioners, especially by district midwives, there are still few published data on the effectiveness of community-oriented cervical screening programs in Greece. OBJECTIVE: To explore an innovative approach in a primary-care setting in rural Crete. This study reports on the effectiveness of a health education meeting in recruiting women for a cervical screening program. METHODS: At a centre for the elderly, 16 women participated in an educational discussion meeting organised to promote cervical screening. The women who participated in the discussion meeting were invited to organise a group visit to a rural primary healthcare centre, in order to be screened as a team during the following 15 days. The theoretical model underpinning the development of this intervention was based on the 'social learning theory'. Emphasis was placed up individual and social responsibility regarding cervical cancer. RESULTS: A total number of 48 women (average age 62 years) including all those who participated in the educational discussion meeting, visited the Spili Health Center, Crete, 15 days after the discussion meeting. The main finding was the high compliance rate of the women who participated in the educational discussion meeting, resulting in recruiting all the initial participants. Moreover, the lay women who participated in the small group discussion meeting, in the capacity of key messengers convinced an additional 32 women to participate in a screening program for cervical cancer as members of a team, rather than individually. The majority (52.1%, n = 25 out of 48) of the women had not been previously screened for cervical cancer. CONCLUSIONS: Health education is an important factor in the process of health promotion, and health professionals should consider the dynamics of a specific group in order to carry out their work effectively. Awareness of the special characteristics of an individual as a member of a defined group can contribute to increasing the motivation for participation in health programs as a member of that group. It is suggested that more educational programs take place in rural Crete in order to augment the compliance rate of women in cervical screening programs.

Aged↗

PubMed1999

Pelvic restraint effect on lumbar gluteal and hamstring muscle electromyographic activation.

OBJECTIVE: To evaluate the influence of pelvic restraint on electromyographic activation (neural drive) of the lumbar extensor, gluteal, and hamstring muscles during submaximal dynamic lumbar extension in an upright seated position. DESIGN: Randomized, balanced, crossover trial comparing the electromyographic activation of the lumbar, gluteal, and hamstring muscles during dynamic lumbar extension exercise with and without pelvic restraint. SETTING: Research laboratory at a private research university. SUBJECTS: Twelve apparently healthy men (ages 18 to 50 yrs). INTERVENTION: Dynamic lumbar extension exercises. MAIN OUTCOME MEASURE: Electromyographic recordings. RESULTS: Pelvic restraint did not influence the relative activation of the lumbar, gluteal, or hamstring muscle groups (p< or =.05) during submaximal dynamic lumbar extension exercise. The relative activation of the gluteal muscles was significantly lower than that of the lumbar extensor and hamstring muscle groups. CONCLUSIONS: Pelvic restraint is not a requirement to promote neural drive to the lumbar musculature during submaximal dynamic lumbar extension exercise performed in an upright seated position. Submaximal lumbar extension exercise reduces the relative contribution of the gluteal muscles compared with a maximal voluntary isometric contraction.

Adult↗