Illuminating the functional and structural repertoire of human TBC/RABGAPs.

The Tre2-Bub2-Cdc16 (TBC) domain-containing RAB-specific GTPase-activating proteins (TBC/RABGAPs) are characterized by the presence of highly conserved TBC domains and act as negative regulators of RABs. The importance of TBC/RABGAPs in the regulation of specific intracellular trafficking routes is now emerging, as is their role in different diseases. Importantly, TBC/RABGAPs act as key regulatory nodes, integrating signalling between RABs and other small GTPases and ensuring the appropriate retrieval, transport and delivery of different intracellular vesicles.

IL-6 Trans-signaling Controls Liver Regeneration After Partial Hepatectomy.

Interleukin-6 (IL-6) is critically involved in liver regeneration after partial hepatectomy (PHX). Previous reports suggest that IL-6 trans-signaling through the soluble IL-6/IL-6R complex is involved in this process. However, the long-term contribution of IL-6 trans-signaling for liver regeneration after PHX is unknown. PHX-induced generation of the soluble IL-6R by ADAM (a disintegrin and metallo) proteases enables IL-6 trans-signaling, in which IL-6 forms an agonistic complex with the soluble IL-6 receptor (sIL-6R) to activate all cells expressing the signal-transducing receptor chain glycoprotein 130 (gp130). In contrast, without activation of ADAM proteases, IL-6 in complex with membrane-bound IL-6R and gp130 activates classic signaling. Here, we describe the generation of IL-6 trans-signaling mice, which exhibit boosted IL-6 trans-signaling and abrogated classic signaling by genetic conversion of all membrane-bound IL-6R into sIL-6R proteins phenocopying hyperactivation of ADAM-mediated shedding of IL-6R as single substrate. Importantly, although IL-6R deficient mice were strongly affected by PHX, survival and regeneration of IL-6 trans-signaling mice was indistinguishable from control mice, demonstrating that IL-6 trans-signaling fully compensates for disabled classic signaling in liver regeneration after PHX. Moreover, we monitored the long-term consequences of global IL-6 signaling inhibition versus IL-6 trans-signaling selective blockade after PHX by IL-6 monoclonal antibodies and soluble glycoprotein 130 as fragment crystallizable fusion, respectively. Both global IL-6 blockade and selective inhibition of IL-6 trans-signaling results in a strong decrease of overall survival after PHX, accompanied by decreased signal transducer and activator of transcription 3 phosphorylation and proliferation of hepatocytes. Mechanistically, IL-6 trans-signaling induces hepatocyte growth factor production by hepatic stellate cells. Conclusion: IL-6 trans-signaling, but not classic signaling, controls liver regeneration following PHX.

Finding acceptable parameter regions of stochastic Hill functions for multisite phosphorylation mechanism.

Multisite phosphorylation plays an important role in regulating switch-like protein activity and has been used widely in mathematical models. With the development of new experimental techniques and more molecular data, molecular phosphorylation processes emerge in many systems with increasing complexity and sizes. These developments call for simple yet valid stochastic models to describe various multisite phosphorylation processes, especially in large and complex biochemical networks. To reduce model complexity, this work aims at simplifying the multisite phosphorylation mechanism by a stochastic Hill function model. Furthermore, this work optimizes regions of parameter space to match simulation results from the stochastic Hill function with the distributive multisite phosphorylation process. While traditional parameter optimization methods have been focusing on finding the best parameter vector, in most circumstances, modelers would like to find a set of parameter vectors that generate similar system dynamics and results. This paper proposes a general α-ß-γ rule to return an acceptable parameter region of the stochastic Hill function based on a quasi-Newton stochastic optimization algorithm. Different objective functions are investigated characterizing different features of the simulation-based empirical data, among which the approximate maximum log-likelihood method is recommended for general applications. Numerical results demonstrate that with an appropriate parameter vector value, the stochastic Hill function model depicts the multisite phosphorylation process well except the initial (transient) period.

Influence of resistance training and herbal supplementation on plasma apelin and metabolic syndrome risk factors in postmenopausal women.

Purpose ­: Menopause is a normal condition that all women experience as they age. The aim of this study was to assess the effects of circuit resistance exercise training with Zataria multiflora (Avishan-e-Shirazi) dietary supplementation on plasma apelin, glucose, insulin, insulin sensitivity and insulin resistance. Methods ­: Ninety-six volunteer postmenopausal women were allocated into 8 groups. Resistance training consisted of 12 stations had been done for 8 weeks. Blood samples were collected at pre- and -post intervention. Results ­: Circuit RT increases apelin and decreases both insulin and blood glucose, whereas Zataria multiflora has no independent effect on apelin but does decrease blood glucose and is likely to be in some means synergistic with circuit RT effects. Conclusion ­: These results suggest that circuit resistance training augments plasma apelin and decreases both insulin and blood glucose. However, Zataria multiflora has no independent effect on apelin but does decrease blood glucose which is likely to be to some extent synergistic with training effects.

Objectif ­: La ménopause est une situation physiologique qui affecte toutes les femmes autour de la cinquantaine. Le but de cette étude était d'évaluer les effets d'un entraînement en résistance de type « circuit training ¼ avec supplémentation orale en Zataria multiflora (Avishan-e-Shirazi) sur l'apeline plasmatique, le glucose, l'insuline, la sensibilité à l'insuline et la résistance à l'insuline. Méthodologie ­: Quatre-vingt quatre femmes postménopausées bénévoles ont été réparties en 8 groupes. L'entraînement en circuit training comprenait 12 séances sur 8 semaines. Des échantillons sanguins ont été prélevés avant et après intervention. Résultats ­: Le circuit training augmente l'apeline et diminue à la fois l'insuline et la glycémie, alors que Zataria multiflora n'a pas d'effet indépendant sur l'apeline, mais diminue la glycémie et est semble synergique des effets du circuit training. Conclusion ­: Ces résultats suggèrent que le circuit training augmente l'apeline plasmatique et diminue insuline et glucose dans le sang. Cependant, Zataria multiflora n'a pas d'effet indépendant sur l'apeline mais fait diminuer la glycémie, ce qui est susceptible d'être dans une certaine mesure synergique avec les effets de l'entraînement.

The balance of interleukin (IL)-6, IL-6·soluble IL-6 receptor (sIL-6R), and IL-6·sIL-6R·sgp130 complexes allows simultaneous classic and trans-signaling.

Interleukin (IL-)6 is the major pro-inflammatory cytokine within the IL-6 family. IL-6 signals via glycoprotein 130 (gp130) and the membrane-bound or soluble IL-6 receptor (IL-6R), referred to as classic or trans-signaling, respectively. Whereas inflammation triggers IL-6 expression, eventually rising to nanogram/ml serum levels, soluble IL-6R (sIL-6R) and soluble gp130 (sgp130) are constitutively present in the upper nanogram/ml range. Calculations based on intermolecular affinities have suggested that systemic IL-6 is immediately trapped in IL-6·sIL-6R and IL-6·sIL-6R·sgp130 complexes, indicating that sIL-6R and sgp130 constitute a buffer system that increases the serum half-life of IL-6 or restricts systemic IL-6 signaling. However, this scenario has not been experimentally validated. Here, we quantified IL-6·sIL-6R and IL-6·sIL-6R·sgp130 complexes over a wide concentration range. The amounts of IL-6 used in this study reflect concentrations found during active inflammatory events. Our results indicated that most IL-6 is free and not complexed with sIL-6R or sgp130, indicating that the level of endogenous sgp130 in the bloodstream is not sufficient to block IL-6 trans-signaling via sIL-6R. Importantly, addition of the single-domain antibody VHH6, which specifically stabilizes IL-6·sIL-6R complexes but did not bind to IL-6 or sIL-6R alone, drove free IL-6 into IL-6·sIL-6R complexes and boosted trans-signaling but not classic signaling, demonstrating that endogenous sIL-6R has at least the potential to form complexes with IL-6. Our findings indicate that even though high concentrations of sIL-6R and sgp130 are present in human serum, the relative ratio of free IL-6 to IL-6·sIL-6R allows for simultaneous classic and trans-signaling.

Micromechanical modeling of rate-dependent behavior of Connective tissues.

In this paper, a constitutive and micromechanical model for prediction of rate-dependent behavior of connective tissues (CTs) is presented. Connective tissues are considered as nonlinear viscoelastic material. The rate-dependent behavior of CTs is incorporated into model using the well-known quasi-linear viscoelasticity (QLV) theory. A planar wavy representative volume element (RVE) is considered based on the tissue microstructure histological evidences. The presented model parameters are identified based on the available experiments in the literature. The presented constitutive model introduced to ABAQUS by means of UMAT subroutine. Results show that, monotonic uniaxial test predictions of the presented model at different strain rates for rat tail tendon (RTT) and human patellar tendon (HPT) are in good agreement with experimental data. Results of incremental stress-relaxation test are also presented to investigate both instantaneous and viscoelastic behavior of connective tissues.

Specific mosaic KRAS mutations affecting codon 146 cause oculoectodermal syndrome and encephalocraniocutaneous lipomatosis.

Oculoectodermal syndrome (OES) and encephalocraniocutaneous lipomatosis (ECCL) are rare disorders that share many common features, such as epibulbar dermoids, aplasia cutis congenita, pigmentary changes following Blaschko lines, bony tumor-like lesions, and others. About 20 cases with OES and more than 50 patients with ECCL have been reported. Both diseases were proposed to represent mosaic disorders, but only very recently whole-genome sequencing has led to the identification of somatic KRAS mutations, p.Leu19Phe and p.Gly13Asp, in affected tissue from two individuals with OES. Here we report the results of molecular genetic studies in three patients with OES and one with ECCL. In all four cases, Sanger sequencing of the KRAS gene in DNA from lesional tissue detected mutations affecting codon 146 (p.Ala146Val, p.Ala146Thr) at variable levels of mosaicism. Our findings thus corroborate the evidence of OES being a mosaic RASopathy and confirm the common etiology of OES and ECCL. KRAS codon 146 mutations, as well as the previously reported OES-associated alterations, are known oncogenic KRAS mutations with distinct functional consequences. Considering the phenotype and genotype spectrum of mosaic RASopathies, these findings suggest that the wide phenotypic variability does not only depend on the tissue distribution but also on the specific genotype.

The relationship between playing computer or video games with mental health and social relationships among students in guidance schools, Kermanshah.

Computer or video games are a popular recreational activity and playing them may constitute a large part of leisure time. This cross-sectional study aimed to evaluate the relationship between playing computer or video games with mental health and social relationships among students in guidance schools in Kermanshah, Islamic Republic of Iran, in 2012. Our total sample was 573 students and our tool was the General Health Questionnaire (GHQ-28) and social relationships questionnaires. Survey respondents reported spending an average of 71.07 (SD 72.1) min/day on computer or video games. There was a significant relationship between time spent playing games and general mental health (P < 0.04) and depression (P < 0.03). There was also a significant difference between playing and not playing computer or video games with social relationships and their subscales, including trans-local relationships (P < 0.0001) and association relationships (P < 0.01) among all participants. There was also a significant relationship between social relationships and time spent playing games (P < 0.02) and its dimensions, except for family relationships.

Numerical simulation of nanoneedle-cell membrane collision: minimum magnetic force and initial kinetic energy for penetration.

Aims and objectives: This research aims to develop a kinetic model that accurately captures the dynamics of nanoparticle impact and penetration into cell membranes, specifically in magnetically-driven drug delivery. The primary objective is to determine the minimum initial kinetic energy and constant external magnetic force necessary for successful penetration of the cell membrane.Model Development: Built upon our previous research on quasi-static nanoneedle penetration, the current model development is based on continuum mechanics. The modeling approach incorporates a finite element method and explicit dynamic solver to accurately represent the rapid dynamics involved in the phenomenon. Within the model, the cell is modeled as an isotropic elastic shell with a hemiellipsoidal geometry and a thickness of 200 nm, reflecting the properties of the lipid membrane and actin cortex. The surrounding cytoplasm is treated as a fluid-like Eulerian body.Scenarios and Results: This study explores three distinct scenarios to investigate the penetration of nanoneedles into cell membranes. Firstly, we examine two scenarios in which the particles are solely subjected to either a constant external force or an initial velocity. Secondly, we explore a scenario that considers the combined effects of both parameters simultaneously. In each scenario, we analyze the critical values required to induce membrane puncture and present comprehensive diagrams illustrating the results.Findings and significance: The findings of this research provide valuable insights into the mechanics of nanoneedle penetration into cell membranes and offer guidelines for optimizing magnetically-driven drug delivery systems, supporting the design of efficient and targeted drug delivery strategies.

Numerical investigation of force and deflection of nanoneedle penetration into cell using finite element approach: Parameter study and experimental validation of results.

This paper aims to develop a numerical methodology to investigate the penetration process of nanoneedles into cells and the corresponding force and indentation length. The finite element approach via the explicit dynamic method handles convergence difficulties in the nonlinear phenomenon. The cell is modeled as an isotropic elastic hemiellipsoidal shell with a thickness of 200 nm, which represents the lipid membrane and actin cortex, encapsulating cytoplasm that is regarded as an Eulerian body because of its fluid-type behavior. Nanoneedles with diameters 400, 200, and 50 nm are considered for model development based on available experimental data. The Von Mises strain failure criterion is used for rupture detection. A parameter study using 1, 2.5, 5, 7.5, and 10 kPa shows that Young's modulus of the HeLa cell membrane is about 5 kPa. Moreover, a failure strain of 1.2 chosen among 0.2, 0.4, 0.6, 0.8, 1, and 1.2 matches best the experimental data. In addition, a diameter study shows that the relations between force-diameter and indentation length-diameter are linear and polynomial, respectively. Furthermore, regarding the experimental data and by using contour of minimum principal stress around needle and an analytical equation for calculation of buckling force of a woven fabric, we proposed that for a given cell, membrane structural stability-a function of the coupled effect of Young's modulus and actin meshwork size-contributes directly to needle insertion success rate for that type of cell.

Trimming of sheep spinal cord by waterjet; an experimental study.

The spinal cord is a structure of nervous tissue that primarily transmits nerve signals from the motor cortex to the body and from the afferent fibers of the sensory neurons to the sensory cortex. It is enveloped by three layers of meninges. Covering provides a supportive framework for the cerebral and cranial vasculature and protects the central nervous system (CNS) from mechanical damage. Surgical operation in the vicinity of the spinal cord is complicated and risky because it exposes it to probably irreversible damage. To reduce the risk of these operations, attempts have been made to remove the tumor using safer methods like waterjet operation. In these methods, the waterjet and spinal cord interaction are inevitable. To secure interaction of operation, a standard development of waterjet criteria is necessary. In this study, a system of waterjet is designed to perform sheep spinal cord as a tissue with a good resemblance to the human spinal cord. Effects of interaction between waterjet and sheep spinal cord are investigated to define a safe operation threshold. The impact of the liquid density of waterjet on failure criteria of spinal cord surgery is also investigated. Results show that meninges are stiff enough to protect the sheep spinal cord from rupture for pressures up to 8 bar; however internal spinal cord tissue cannot be guaranteed any damage. Three essential parameters represent the spinal cord meninges and spinal cord deformation during the tests. These parameters lead us to provide standard criteria for damage prevention of the spinal cord.

Two novel germline KRAS mutations: expanding the molecular and clinical phenotype.

Noonan and Cardio-facio-cutaneous (CFC) syndromes are characterized by typical dysmorphic features, cardiac defects, short stature, variable ectodermal anomalies, and intellectual disability. Both belong to the Ras/mitogen-activated protein kinase pathway group of disorders and clinical features overlap other related conditions, notably LEOPARD and Costello syndromes. KRAS mutations account for about 2% of reported Noonan and <5% of reported CFC cases. The mutation spectrum includes recurrent missense changes clustering in particular domains of the KRAS protein and conferring gain-of-function. We report three patients from two unrelated families with novel missense KRAS mutations, p.K147E and p.Y71H. Both mutations affect a residue which is highly conserved in KRAS and other RAS isoforms. One of the families includes a mother and son pair who represent the first report of a vertically transmitted KRAS mutation. In addition, the mother and son pair had peripheral neuropathy, complicated by Charcot arthropathy in the mother. An unusual phenotypic effect of the specific KRAS mutation or a coincidence of two independent disorders may be considered. KRAS mutation-associated phenotypes appear to be subject to considerable clinical heterogeneity. All three cases highlight the challenges of clinical assessment in KRAS mutation-positive patients, and the utility of molecular testing as an adjunct to diagnosis.

Structural and mechanistic insights into the interaction between Rho and mammalian Dia.

Formins are involved in a variety of cellular processes that require the remodelling of the cytoskeleton. They contain formin homology domains FH1 and FH2, which initiate actin assembly. The Diaphanous-related formins form a subgroup that is characterized by an amino-terminal Rho GTPase-binding domain (GBD) and an FH3 domain, which bind somehow to the carboxy-terminal Diaphanous autoregulatory domain (DAD) to keep the protein in an inactive conformation. Upon binding of activated Rho proteins, the DAD is released and the ability of the formin to nucleate and elongate unbranched actin filaments is induced. Here we present the crystal structure of RhoC in complex with the regulatory N terminus of mammalian Diaphanous 1 (mDia1) containing the GBD/FH3 region, an all-helical structure with armadillo repeats. Rho uses its 'switch' regions for interacting with two subdomains of GBD/FH3. We show that the FH3 domain of mDia1 forms a stable dimer and we also identify the DAD-binding site. Although binding of Rho and DAD on the N-terminal fragment of mDia1 are mutually exclusive, their binding sites are only partially overlapping. On the basis of our results, we propose a structural model for the regulation of mDia1 by Rho and DAD.

A Non-cutaneous Form of Melanoma in a Goat during Meat Inspection: a Case Report.

Malignant melanoma is a neoplasm that originates from melanocytes. This tumor is observed in cutaneous and non-cutaneous forms, and it is considered one of the most life-threatening types of cancers. Non-cutaneous melanoma is a complex of unique and malignant complications that are easily separable from cutaneous type. Since the ultraviolet radiation from the sun damages DNA and is an oxidative stress factor in melanoma and there are more melanocytes in the basal layer of skin than other parts of the body, the cutaneous form has more prevalence. Most of the time, non-cutaneous form is the result of cutaneous metastasis but both forms can occur primarily. Furthermore, non-cutaneous form usually happens in mucosal layers, intestines, and eyes; moreover, the main reasons are ectopic melanocytes or their unwanted regressive growing. Malignant melanoma can occur in all domestic animals; however, they seem to be rare in sheep and goats. Herein, we describe a rare case of the primary non-cutaneous form of malignant melanoma in a three-year-old indigenous female goat. During meat inspection procedures in a slaughterhouse in Tabriz, Iran, we encountered numerous round firm black masses on visceral surfaces and serous membranes of the abdominal and thoracic cavities. The liver and lungs were prominently affected. Samples were taken from involved parts, and malignant melanoma was confirmed in the histopathological examination due to pleomorphism and polymorphism and melanin pigments in cells with eosinophilic cytoplasm. According to what was stated in the &quot;manual on meat inspection for developing countries&quot;, the carcass was not convenient for human use and condemned by the inspector.

Modeling, simulation, and optimal initiation planning for needle insertion into the liver.

Needle insertion simulation and planning systems (SPSs) will play an important role in diminishing inappropriate insertions into soft tissues and resultant complications. Difficulties in SPS development are due in large part to the computational requirements of the extensive calculations in finite element (FE) models of tissue. For clinical feasibility, the computational speed of SPSs must be improved. At the same time, a realistic model of tissue properties that reflects large and velocity-dependent deformations must be employed. The purpose of this study is to address the aforementioned difficulties by presenting a cost-effective SPS platform for needle insertions into the liver. The study was constrained to planar (2D) cases, but can be extended to 3D insertions. To accommodate large and velocity-dependent deformations, a hyperviscoelastic model was devised to produce an FE model of liver tissue. Material constants were identified by a genetic algorithm applied to the experimental results of unconfined compressions of bovine liver. The approach for SPS involves B-spline interpolations of sample data generated from the FE model of liver. Two interpolation-based models are introduced to approximate puncture times and to approximate the coordinates of FE model nodes interacting with the needle tip as a function of the needle initiation pose; the latter was also a function of postpuncture time. A real-time simulation framework is provided, and its computational benefit is highlighted by comparing its performance with the FE method. A planning algorithm for optimal needle initiation was designed, and its effectiveness was evaluated by analyzing its accuracy in reaching a random set of targets at different resolutions of sampled data using the FE model. The proposed simulation framework can easily surpass haptic rates (>500 Hz), even with a high pose resolution level ( approximately 30). The computational time required to update the coordinates of the node at the needle tip in the provided example was reduced from 177 s to 0.8069 ms. The planning accuracy was acceptable even with moderate resolution levels: root-mean-square and maximum errors were 1 mm and 1.2 mm, respectively, for a pose and PPT resolution levels of 17 and 20, respectively. The proposed interpolation-based models significantly improve the computational speed of needle insertion simulation and planning, based on the discretized (FE) model of the liver and can be utilized to establish a cost-effective planning platform. This modeling approach can also be extended for use in other surgical simulations.

Formation of a transition-state analog of the Ras GTPase reaction by Ras-GDP, tetrafluoroaluminate, and GTPase-activating proteins.

Unlike the alpha subunits of heterotrimeric guanosine triphosphate (GTP)-binding proteins, Ras-related GTP-binding proteins have hitherto been considered not to bind or become activated by tetrafluoroaluminate (AIF4-). However, the product of the proto-oncogene ras in its guanosine diphosphate (GDP)-bound form interacted with AIF4 - in the presence of stoichiometric amounts of either of the guanosine triphosphatase (GTPase)-activating proteins (GAPs) p120GAP and neurofibromin. Neither oncogenic Ras nor a GAP mutant without catalytic activity produced such a complex. Together with the finding that the Ras-binding domain of the protein kinase c-Raf, whose binding site on Ras overlaps that of the GAPs, did not induce formation of such a complex, this result suggests that GAP and neurofibromin stabilize the transition state of the GTPase reaction of Ras.

The Ras-RasGAP complex: structural basis for GTPase activation and its loss in oncogenic Ras mutants.

The three-dimensional structure of the complex between human H-Ras bound to guanosine diphosphate and the guanosine triphosphatase (GTPase)-activating domain of the human GTPase-activating protein p120GAP (GAP-334) in the presence of aluminum fluoride was solved at a resolution of 2.5 angstroms. The structure shows the partly hydrophilic and partly hydrophobic nature of the communication between the two molecules, which explains the sensitivity of the interaction toward both salts and lipids. An arginine side chain (arginine-789) of GAP-334 is supplied into the active site of Ras to neutralize developing charges in the transition state. The switch II region of Ras is stabilized by GAP-334, thus allowing glutamine-61 of Ras, mutation of which activates the oncogenic potential, to participate in catalysis. The structural arrangement in the active site is consistent with a mostly associative mechanism of phosphoryl transfer and provides an explanation for the activation of Ras by glycine-12 and glutamine-61 mutations. Glycine-12 in the transition state mimic is within van der Waals distance of both arginine-789 of GAP-334 and glutamine-61 of Ras, and even its mutation to alanine would disturb the arrangements of residues in the transition state.

GTPase-activating proteins: helping hands to complement an active site.

Stimulation of the intrinsic GTPase activity of GTP-binding proteins by GTPase-activating proteins (GAPs) is a basic principle of GTP-binding-protein downregulation. Recently, the molecular mechanism behind this reaction has been elucidated by studies on Ras and Rho, and their respective GAPs. The basic features involve stabilizing the existing catalytic machinery and supplementing it by an external arginine residue. This represents a novel mechanism for enzyme active-site formation.

Time-dependent analysis of leaflets in mechanical aortic bileaflet heart valves in closing phase using the finite strip method.

BACKGROUND AND AIMS OF THE STUDY: Mechanical heart valves (MHV) are widely used to replace dysfunctional and failed heart valves. The bileaflet MHV design is very popular due to its superior hemodynamics. Since their introduction in 1977, the hemodynamics of bileaflet prostheses has been extensively studied. In this study the dynamic behaviour during the closing phase of a bileaflet MHV under normal physiological conditions has been investigated. METHODS: Fluid analysis is based on the control volume with moving boundaries in the vicinity of the occluder. Unsteady continuity equation, unsteady momentum equation on the control volume and unsteady Bernoulli's equation have been used to calculate velocity of blood flow and force on the occluder tip. To solve the governing equations for the calculation of pressure and the related force, the finite strips method has been implemented. Only 32 strips are sufficient to calculate the force due to pressure on the leaflets. The equations of motion have been solved using the Runge-Kutta method in the fourth order. RESULTS: The maximum velocity of the leakage flow in the closing phase falls within the range of 3.5-4.4 m/s. The maximum velocity of the occluder tip is in the range of 2.4-3.2 m/s. The backflow also exhibits oscillation similar to that of the occluder with net backflow rate in the range of 9.7-12.3 ml/beat. The impact force between occluder and its housing is in the range of 80-140 N and impact occurs during 33.1-41.0 ms and the leaflets are completely settled at 108-115 ms in the closing phase. CONCLUSION: The finite strip method was implemented to study the closing phase of a bileaflet MHV. Results are consistent with the previous experimental data. This method is of general applicability to study dynamic behaviour of MHVs.

Abnormalities of fragile histidine triad genomic and complementary DNAs in cervical cancer: association with human papillomavirus type.

BACKGROUND: Chromosome 3p14.2 contains FRA3B, the most active chromosome breakage site in the human genome. The fragile histidine triad (FHIT) gene, a putative tumor suppressor gene, overlaps FRA3B. Human papillomavirus (HPV), a known cofactor in cervical carcinogenesis, can integrate into FRA3B. We examined abnormalities in FHIT and its RNA transcripts in cervical cancer cell lines and tumors. We also investigated the relationship between loss of heterozygosity (LOH) in FHIT/FRA3B and the presence of oncogenic HPV types. METHODS: Eleven cell lines, 40 tumors (20 fresh and 20 archival), and 10 normal cervical epithelia were examined. Two intragenic polymorphic markers (D3S1300 and D3S4103) and the polymerase chain reaction (PCR) were used to examine FHIT LOH. Reverse transcription-PCR (RT-PCR) analysis and single-strand conformation polymorphism analysis of RT-PCR products were used to characterize FHIT transcripts. Oncogenic HPV types were identified by PCR, using general and type-specific primers. RESULTS: All normal epithelia, 19 of 20 fresh tumors and nine of 11 cell lines expressed wild-type and, occasionally, exon 8-deleted FHIT transcripts. Additional aberrant FHIT transcripts were seen in nine of 20 fresh tumors and in seven of 11 cell lines. DNA sequencing of the aberrant transcripts revealed a variety of insertions and deletions but no point mutations. Three cell lines also had homozygous FHIT deletions. Oncogenic HPV types (i.e., 16, 18, 31, and 33) were detected in 18 of 20 archival tumors, and, in these tumors, LOH within FHIT was identified in nine of 16 informative cases. HPV 16 was found to be associated with LOH in the FHIT/FRA3B region (P = .041). CONCLUSION: FHIT/FRA3B is frequently altered in cervical cancer, demonstrating LOH, occasional homozygous deletions, and frequent aberrant transcripts not found in normal epithelia. However, the presence of wild-type transcripts and the lack of protein-altering point mutations raise questions about FHIT's function as a classic tumor suppressor gene in cervical tissue.