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Ankle eversion torque response to sudden ankle inversion Torque response in unbraced, braced, and pre-activated situations.

In 13 young ankle stable subjects, ankle eversion torque and peroneal EMG were simultaneously recorded in response to sudden ankle inversion. The eversion torque response was bi-phasic. The initial development of torque, which was responsible for 30% of the maximal eversion torque response, was observed 135 ms after the start of platform rotation and correlated well with the onset of the automatic postural peroneal EMG response. The remaining eversion torque response commenced after 305 ms, strongly correlating with the onset of the peroneal long latency voluntary EMG activity. With the ankle unbraced, 66% of the maximal torque level was reached in 326 ms. While braced, the same torque magnitude was reached using 230 ms (p<0.02), and pre-activation of the peroneal muscles allowed the subjects to reach the same level of torque in 89 ms (p<0.0005). Prior to the study, a common reaction pattern to sudden inversion was expected in an ankle stable population, but review of the eversion torque and EMG data from the 13 subjects revealed three different voluntary reaction patterns: 10 subjects showed an efficient activation of evertor muscles; two subjects stiffened their ankles with activation of both in- and evertor muscles; and one subject showed a marginal voluntary activation of the ankle evertors. The results of the study indicate that the reaction to sudden ankle inversion is not solely automatic. The main part of the torque response is voluntarily mediated and inter-individual differences in strategy seem to exist in healthy subjects.

Adult↗

A comprehensive review and analysis of 25 years of data from an in vivo canine model of sudden cardiac death: implications for future anti-arrhythmic drug development.

Sudden cardiac death resulting from ventricular tachyarrhythmias remains the leading cause of death in industrially developed countries, accounting for between 300,000 and 500,000 deaths each year in the United States. Yet, despite the enormity of this problem, the development of safe and effective anti-arrhythmic agents remains elusive. The identification of effective anti-arrhythmic agents is critically dependent upon the use of appropriate animal models of human disease. During the last 25 years, a canine model of sudden cardiac death has proven to be useful in both the identification of factors contributing to ventricular fibrillation (VF) and the evaluation of potential anti-arrhythmic therapies. The present review provides a detailed retrospective analysis of the data obtained with this model. Briefly, VF was reliably and reproducibly induced by the combination of acute myocardial ischemia at site distant from a previous myocardial infarction during submaximal exercise (to activate the autonomic nervous system). This exercise plus ischemia test identified 2 stable populations of dogs: those that development malignant arrhythmias (susceptible, n=303) and those that rarely developed even single premature ventricular activation (resistant, n=209). The susceptible animals exhibited an elevated sympathetic activation (due to an enhanced beta2-adrenoceptor responsiveness) and a subnormal parasympathetic regulation. Several interventions have proven to be particularly effective in preventing VF in the susceptible dogs; including calcium channel antagonists, left stellate ganglion disruption, ATP-sensitive potassium channel antagonists, beta-adrenoceptor antagonists, and non-pharmacological interventions (endurance exercise training and dietary omega-3 fatty acids).

Animals↗

Construction and applications of yellow fever virus replicons.

Subgenomic replicons of yellow fever virus (YFV) were constructed to allow expression of heterologous reporter genes in a replication-dependent manner. Expression of the antibiotic resistance gene neomycin phosphotransferase II (Neo) from one of these YFV replicons allowed selection of a stable population of cells (BHK-REP cells) in which the YFV replicon persistently replicated. BHK-REP cells were successfully used to trans-complement replication-defective YFV replicons harboring large internal deletions within either the NS1 or NS3 proteins. Although replicons with large deletions in either NS1 or NS3 were trans-complemented in BHK-REP, replicons that contained deletions of NS3 were trans-complemented at lower levels. In addition, replicons that retained the N-terminal protease domain of NS3 in cis were trans-complemented with higher efficiency than replicons in which both the protease and helicase domains of NS3 were deleted. To study packaging of YFV replicons, Sindbis replicons were constructed that expressed the YFV structural proteins in trans. Using these Sindbis replicons, both replication-competent and trans-complemented, replication-defective YFV replicons could be packaged into pseudo-infectious particles (PIPs). Although these results eliminate a potential role of either NS1 or full-length NS3 in cis for packaging and assembly of the flavivirus virion, they do not preclude the possibility that these proteins may act in trans during these processes.

Cell Line↗

Djeyes absent (Djeya) controls prototypic planarian eye regeneration by cooperating with the transcription factor Djsix-1.

A conserved network of nuclear proteins is crucial to eye formation in both vertebrates and invertebrates. The finding that freshwater planarians can regenerate eyes without the contribution of Pax6 suggests that alternative combinations of regulatory elements may control the morphogenesis of the prototypic planarian eye. To further dissect the molecular events controlling eye regeneration in planarians, we investigated the role of eyes absent (Djeya) and six-1 (Djsix-1) genes in Dugesia japonica. These genes are expressed in both regenerating eyes and in differentiated photoreceptors of intact adults. Through RNAi studies, we show that Djsix-1 and Djeya are both critical for the regeneration of normal eyes in planarians and genetically cooperate in vivo to establish correct eye cell differentiation. We further demonstrate that the genetic interaction is mediated by physical interaction between the evolutionarily conserved domains of these two proteins. These data indicate that planarians use cooperatively Djsix-1 and Djeya for the proper specification of photoreceptors, implicating that the mechanism involving their evolutionarily conserved domains can be very ancient. Finally, both Djsix-1 and Djeya double-stranded RNA are substantially more effective at producing no-eye phenotypes in the second round of regeneration. This is probably due to the significant plasticity of the planarian model system, based on the presence of a stable population of totipotent stem cells, which ensure the rapid cell turnover of all differentiated cell types.

Amino Acid Sequence↗

Ability of the exercise electrocardiogram test to detect ischemia in stable coronary artery disease patients with ST-segment depression on the resting electrocardiogram.

BACKGROUND: The appearance of ST-segment depression on the exercise electrocardiogram (ETT) generally indicates myocardial ischemia. Disorders that produce ST-segment depression on the resting electrocardiogram can confound the results of the ETT. The purpose of this study was to assess the reliability of exercise ST-segment depression as an indicator of ischemia in patients with a resting electrocardiographic ST depression, presumably caused by coronary disease. METHODS: We studied 882 patients 1 to 6 months after a hospitalization for myocardial infarction or unstable angina as part of the Multicenter Study of Myocardial Ischemia. An ETT and thallium-201 perfusion scan were performed in all patients. For the purpose of this study, the perfusion scan was considered the "gold standard" for ischemia. We correlated the ETT ST-segment responses with the perfusion scan in a group with a normal electrocardiogram baseline ST segment (NO ST DEP group) and in a group with resting electrocardiographic ST depression > or =0.5 mm (ST DEP group). RESULTS: Although the frequency of ischemia by perfusion scan was similar in the ST DEP group (48%) to that in the NO ST DEP group (40%), the ST DEP group had a significantly higher (p = 0.03) sensitivity (54%) in detecting ischemia than the NO ST DEP group (35%). The ST DEP group, however, had a lower (58% vs 74%) specificity than the NO ST DEP group (p = 0.056). Baseline characteristics of the two groups probably account for these differences. The overall diagnostic accuracy is similar (58% vs 56%) in the two groups. CONCLUSION: We conclude that in a stable population with known coronary disease, the presence of ST depression on the resting electrocardiogram does not impair the detection of ischemia by the ETT, but may be associated with a higher false-positive rate.

Adult↗

The single nuclear lamin of Caenorhabditis elegans forms in vitro stable intermediate filaments and paracrystals with a reduced axial periodicity.

The lamins of the tunicate Ciona intestinalis and the nematode Caenorhabditis elegans show unusual sequence features when compared to the more than 35 metazoan lamin sequences currently known. We therefore analyzed the in vitro assembly of these two lamins by electron microscopy using chicken lamin B2 as a control. While lamin dimers usually appear as a rod carrying two globules at one end, these globules are absent from Ciona lamin, which lacks the central 105-residue region of the tail domain. The deletion of 14 residues or two heptads from the coiled coil rod domain of the single C.elegans lamin results in a 1.5-nm shortening of the dimer rod. Similarly, the paracrystals assembled from the C.elegans lamin exhibit a 3.1-nm reduction of the true axial repeat compared to that of chicken lamin B2 paracrystals. We speculate that the banding pattern in the C.elegans lamin paracrystals arises from a relative stagger between dimers and/or a positioning of the globular tail domain relative to the central rod that is distinct from that observed in chicken lamin B2 paracrystals. Here we show that a nuclear lamin can assemble in vitro into 10-nm intermediate filaments (IFs). C.elegans lamin in low ionic strength Tris-buffers at a pH of 7.2-7.4 provides a stable population of lamin IFs. Some implications of this filament formation are discussed.

Amino Acid Sequence↗

Genetic and strategic models for the evolution of mating systems.

Male and female fitnesses in the Shaw-Mohler equation are partitioned into components which putatively determine mating systems. The resultant genetic models provide criteria for evolutionary stable population states and yield strategic models based on maximization principles and fitness sets.

Alleles↗

The human hypothalamo-neurohypophyseal system in relation to development, aging and Alzheimer's disease.

The research reviewed in the present paper indicates that vasopressin and oxytocin cells in the human HNS constitute an extremely stable population of neurons throughout the human life span. Increases in the activity of these cells, which are probably related to maturation of the system were observed during fetal development and probably extend well beyond term. During senescence an increase in the activity of the vasopressin cells in the human HNS was observed which is probably a compensation for age-related changes in kidney function. These data do not support a role of declining vasopressin secretion in age-related memory decline. Although there is some evidence for an impairment of vasopressin synthesis and release in Alzheimer patients, vasopressin cell numbers in Alzheimer's disease do not fall below values observed in young controls. Furthermore, peripheral administration of vasopressin or vasopressin analogues to AD patients have not yielded consistent results.

Aged↗

IFN-gamma-producing effector CD8+ T cells and IL-10-producing regulatory CD4+ T cells in fixed drug eruption.

BACKGROUND: Although effector and regulatory T cells play roles in the progression and resolution of inflammatory diseases, respectively, little in vivo data exist regarding the T-cell dynamics in the pathogenesis of inflammatory skin diseases in humans. OBJECTIVE: Our aim is to phenotypically and functionally characterize the T cells responsible for initiation and regulation of inflammatory events in fixed drug eruption (FDE) as a disease model to study the role of cytokines produced within the epidermis in the pathogenesis of inflammatory skin disease. METHODS: By use of flow cytometry, we phenotypically and functionally characterized the intraepidermal T cells that persist as a stable population in resting (pigmented) FDE lesions and that are present in active FDE lesions. RESULTS: In resting FDE lesions, most of the intraepidermal T cells were of the CD8 phenotype, most of which expressed cutaneous lymphocyte-associated antigen, alpha4beta1, CD11a, alphaEbeta7, and CD45RA but not CD27, CD62L, CCR4, or CCR7. This population selectively expressed CD122 but not CD25. Intracellular staining demonstrated that most intraepidermal CD8+ T cells were capable of producing IFN-gamma and TNF-alpha but produced little IL-2 and IL-4. On the other hand, in the FDE lesions that arose after challenge, a significant number of CD4+ T cells capable of producing IL-10 migrated into the lesional epidermis. Moreover, nearly 70% of the CD4+ T cells migrating into the lesional epidermis expressed CD25. CONCLUSIONS: Effector IFN-gamma-producing CD8+ T cells and regulatory IL-10-producing CD4+ T cells might be responsible for the progression and resolution of FDE, respectively.

Adult↗

Childhood obesity: public-health crisis, common sense cure.

During the past two decades, the prevalence of obesity in children has risen greatly worldwide. Obesity in childhood causes a wide range of serious complications, and increases the risk of premature illness and death later in life, raising public-health concerns. Results of research have provided new insights into the physiological basis of bodyweight regulation. However, treatment for childhood obesity remains largely ineffective. In view of its rapid development in genetically stable populations, the childhood obesity epidemic can be primarily attributed to adverse environmental factors for which straightforward, if politically difficult, solutions exist.

Child↗

Tuberculosis in the Potteries 1971-74.

Notification rates of all forms of tuberculosis have increased in all age-groups in the Potteries, in a stable population which includes only a small immigrant community. The increase is greatest in both males and females over the age of 65 years and under 15 years. High notification rates have been recorded in workers in the pottery and mining industries and were unrelated to pneumoconiosis. Discrepancies have been found between the numbers of notified cases and the numbers of laboratory isolates of tubercle bacilli, particularly in cases of non-respiratory disease. This indicates that not all proven cases of tuberculosis are notified. The study has revealed serious deficiencies in the contact-tracing procedure in certain areas of the Potteries consequent upon the closure and reorganisation of outlying chest clinics.

Adolescent↗

Quaternary structure of bovine alpha-crystallin: influence of temperature.

The tertiary and quaternary structure of alpha-crystallin is still a matter of controversy. We have characterized the native alpha-crystallin quaternary structure by isolating it at the in vivo temperature and solvent conditions. It can be represented by a distribution of expanded particles with a weight average molar mass of 550,000 g/mol. On decreasing (to 4 degrees C) or increasing (up to 50 degrees C) the temperature, the size distribution increases to larger particles. Only at lower temperatures (4 degrees C), a stable population of particles is obtained with weight average molar mass of 700,000 g/mol. In all conditions, alpha-crystallin behaves as a very expanded particle with a maximum hydrodynamic volume of 3.15 ml/g. The transitions in quaternary structure are rather slow: it takes several hours to evolve from a population of aggregates, characteristic for given solvent conditions, to another distribution in size and quaternary structure on changing the environment. The quaternary structure of alpha-crystallin is an uncharacteristic parameter of the particle: a broad distribution of values can be obtained on changing the environment. Any realistic model should include this property. Our studies favor an open loose structure, where peptides can be added or removed without drastic changes of secondary and tertiary structure of the peptides.

Animals↗

Heterogeneity in microparticle formation and exposure of anionic phospholipids at the plasma membrane of single adherent platelets.

Adherent platelets were examined for their ability to form microvesicles and procoagulant sites for thrombin formation. Epifluorescence and phase-contrast microscopy were employed to visualize shape changes, changes in intracellular Ca(2+) levels ([Ca(2+)](i)), vesiculation of the plasma membrane and appearance of anionic phospholipids in the outer leaflet of the plasma membrane, as probed by annexin V binding. In the absence of extracellular Ca(2+) two stable populations of adherent platelets were observed. The majority of the adherent platelets were fully spread and about 10% remained in a non-spread dendritic state. In the presence of extracellular Ca(2+) vesiculation at the surface of spread platelets occurred at a rather slow rate (10% of the platelets after 20 min) concomitantly with an increase in [Ca(2+)](i) and binding of annexin V. However, a small fraction of the adherent platelets ( approximately 1%) responded much faster. Ionomycin-enhanced influx of Ca(2+) in dendritic platelets resulted in a rapid transformation of these platelets into inflated, balloon-shaped, platelets having a diameter of 2.0+/-0.7 microm without notable microvesicle formation. In contrast, fully spread platelets retained their shape but obtained frayed edges as a result of microvesicle formation. Confocal scanning fluorescence microscopy indicated that annexin V bound to very distinct sites at the outer plasma membrane of spread as well as balloon-shaped platelets. Inhibition of platelet calpain activity suppressed ionomycin-enhanced microvesicle formation and ballooning of platelets, but not annexin V binding. These findings indicate that vesiculation and ballooning, but not the exposure of phosphatidylserine at the outer leaflet of the adherent platelet membrane, are associated with cytoskeleton destruction. Altogether, the data suggest a similar relationship between [Ca(2+)](i) and the formation of platelet procoagulant sites as reported for platelets in suspension. However, the present investigations on single adherent platelets reveal for the first time that adhesion and spreading of platelets is not necessarily associated with the appearance of procoagulant sites. Secondly, an unexpected diversity was observed among adherent platelets with respect to sensitivity to Ca(2+)-induced generation of procoagulant sites and Ca(2+)-induced vesiculation of plasma membrane. It is tempting to speculate that this diversity is of importance for the procoagulant response of platelets to a hemostatic challenge elicited by an injured vessel wall.

Anions↗

Fusion protein predicted amino acid sequence of the first US avian pneumovirus isolate and lack of heterogeneity among other US isolates.

Avian pneumovirus (APV) was first isolated from turkeys in the west-central US following emergence of turkey rhinotracheitis (TRT) during 1996. Subsequently, several APV isolates were obtained from the north-central US. Matrix (M) and fusion (F) protein genes of these isolates were examined for sequence heterogeneity and compared with European APV subtypes A and B. Among US isolates the M gene shared greater than 98% nucleotide sequence identity with only one nonsynonymous change occurring in a single US isolate. Although the F gene among US APV isolates shared 98% nucleotide sequence identity, nine conserved substitutions were detected in the predicted amino acid sequence. The predicted amino acid sequence of the US APV isolate's F protein had 72% sequence identity to the F protein of APV subtype A and 71% sequence identity to the F protein of APV subtype B. This compares with 83% sequence identity between the APV subtype A and B predicted amino acid sequences of the F protein. The US isolates were phylogenetically distinguishable from their European counterparts based on F gene nucleotide or predicted amino acid sequences. Lack of sequence heterogeneity among US APV subtypes indicates these viruses have maintained a relatively stable population since the first outbreak of TRT. Phylogenetic analysis of the F protein among APV isolates supports classification of US isolates as a new APV subtype C.

Amino Acid Sequence↗

Absence of nerve-dependent conversion of rapidly degrading to stable acetylcholine receptors at adult innervated endplates.

It has been suggested that acetylcholine receptors newly inserted into adult innervated endplates have a rapid degradation rate, but are normally converted to a stable, slowly degrading form in a nerve-dependent fashion. Denervation therefore should eliminate conversion and cause pre-existing unconverted receptors to continue degrading rapidly. We tested this model of nerve-dependent conversion in mouse sternomastoid muscle, using quantitative electron microscopic autoradiography in order to specifically examine degradation of receptors at identified endplate membrane. Prior to denervation, we labelled the receptors with sequential alpha-bungarotoxin exposures, using conditions designed to maximize the predicted effect of denervation. However, we observed no difference in the rate of receptor degradation at innervated and denervated endplates up to seven days after denervation (at which time accelerated degradation of pre-existing stabilized receptors is known to begin in this muscle). The regulation of endplate acetylcholine receptor metabolic turnover is a complex and still largely undefined issue, related to many factors such as subunit composition, cytoskeleton and basement membrane composition, muscle activity, and neural influences. In particular, the nerve's influence on the normal stabilization of receptors at innervated adult endplates has been controversial. Our data indicate that slow degradation is probably an inherent property of newly inserted junctional receptors, and argue against nerve-dependent conversion and stabilization. Based on the present data, however, we cannot rule out the presence of a small nerve-independent subpopulation that degrades rapidly. The molecular mechanisms involved in establishing and maintaining a stable population of adult endplate acetylcholine receptors remain to be established.

Animals↗

Cessation of reproduction: an analytic view of menopause.

While there is evidence for genetic control of ovarian and follicular development, only recently have reports appeared on genetic components involved in ovarian failure. Unlike predictions of a stable population, age at menarche is decreasing, while female life span is increasing. This leads to examination of genotype-environment interactions. Evolution of the large human brain size has been accompanied by a reduction in size of the gastrointestinal tract. Consequences, in terms of altered diet and effects on menarche/menstruation/menopause were discussed. In the earliest days of human evolution, genotoxic agents and marginal diets could have produced heritable changes in cessation of menstruation. Toxic alterations may still be apparent, such as epidemiological studies on the effects of smoking on age at menopause. Attempts to reconstruct some of the recent past history, from coprolites to findings in frozen human specimens, have to be extended still further into the past.

Environment↗

Coeliac disease: effect of early feeding on the incidence of the disease.

Coeliac disease, also called permanent gluten sensitive enteropathy, has recently been recognised as constituting a widespread health problem. Effective treatment involves the strict exclusion of wheat, rye, barley and possibly also oats from the diet. Genetic susceptibility and the presence of gluten in the diet are prerequisites for developing the disease. Sweden has recently experienced an epidemic of coeliac disease in children below 2 years of age. Previously, considerable changes in incidence over time have also been reported from England, Scotland and Ireland. Such obvious changes in incidence over rather short time periods, in genetically stable populations, emphasise the importance of environmental factors in the aetiology. Thus, most likely, the aetiology of coeliac disease is multifactorial. However, further conclusive evidence is required to settle if environmental factors, beyond presence of gluten in the diet, really influence the immunological process resulting in the coeliac small intestinal lesion, or merely influence the clinical expression of the disease. The search for contributing exposures has thus far focused on early feeding, suggesting that breast-feeding duration and the amount of gluten consumed are of importance, and possibly also the age for introducing gluten into the diet of infants. The pattern of causation may vary over time and between countries, however, which may obscure the search for risk factors. Nevertheless, a challenging possibility that needs to be explored is if coeliac disease can be delayed, or possibly even prevented for an entire life span, by favourable dietary habits.

Breast Feeding↗

A prospective, blinded determination of the natural history of aspirin resistance among stable patients with cardiovascular disease.

OBJECTIVES: This study was designed to determine if aspirin resistance is associated with clinical events. BACKGROUND: Aspirin resistance, defined by platelet function testing and presumed clinical unresponsiveness to aspirin, has been previously reported by our group and others. However, little information exists linking the laboratory documentation of aspirin resistance and long-term clinical events. METHODS: We prospectively enrolled 326 stable cardiovascular patients from 1997 to 1999 on aspirin (325 mg/day for > or =7 days) and no other antiplatelet agents. We tested for aspirin sensitivity by optical platelet aggregation using adenosine diphosphate (ADP) and arachidonic acid (AA). The primary outcome was the composite of death, myocardial infarction (MI), or cerebrovascular accident (CVA). Mean follow-up was 679 +/- 185 days. Aspirin resistance was defined as a mean aggregation of > or =70% with 10 microM ADP and > or =20% with 0.5 mg/ml AA. RESULTS: Of the patients studied, 17 (5.2%) were aspirin resistant and 309 (94.8%) were not aspirin resistant. During follow-up, aspirin resistance was associated with an increased risk of death, MI, or CVA compared with patients who were aspirin sensitive (24% vs. 10%, hazard ratio [HR] 3.12, 95% confidence interval [CI] 1.10 to 8.90, p = 0.03). Stratified multivariate analyses identified platelet count, age, heart failure, and aspirin resistance to be independently associated with major adverse long-term outcomes (HR for aspirin resistance 4.14, 95% CI 1.42 to 12.06, p = 0.009). CONCLUSIONS: This study demonstrates the natural history of aspirin resistance in a stable population, documenting a greater than threefold increase in the risk of major adverse events associated with aspirin resistance.

Adult↗