Fibrinogen biosynthesis in isolated guinea pig megakaryocytes

Fibrinogen synthesis was investigated in guinea pig megakaryocytes. Purified megakaryocytes were incubated with 35S-methionine in methionine-free incubation medium for 18 hours. Newly synthesized fibrinogen in megakaryocyte lysates enriched with purified carrier guinea pig fibrinogen was immunoprecipitated with a specific anti- guinea pig fibrinogen antiserum produced in rabbits. Proteins in the immunoprecipitates were analyzed with a 3.5% to 10.0% gradient polyacrylamide slab gel electrophoresis and auto-radiography. Radioactivity was detected in a protein band of 340,000 daltons. In order to verify fibrinogen synthesis, immunoprecipitate was analyzed by two-dimensional slab gel electrophoresis: (1) the first dimension separated unreduced fibrinogen using a 3.5% to 10.0% gradient gel; (2) following reduction by 2-beta-mercaptoethanol, fibrinogen chains were separated in the second dimension using a 10% gel. Alpha, beta, and gamma fibrinogen chains, which represented carrier guinea pig plasma fibrinogen, were visualized by Coomassie brilliant blue. Autoradiography identified the incorporation of radioactivity into the three fibrinogen chains. In control experiments, immunoprecipitates, produced by exposing megakaryocyte lysates to preimmune rabbit serum and goat anti-rabbit IgG, were also analyzed by the two-dimensional gel system. Radioactivity was not detected in sites corresponding to the migration of fibrinogen subunits. The study demonstrates that isolated guinea pig megakaryocytes can synthesize fibrinogen. The electrophoretic mobility of newly synthesized fibrinogen and subunits is similar to that of guinea pig plasma fibrinogen.     

Long-term adaptation to prism-induced inversion of the retinal images

For 1 week, healthy human participants ( n=7) were devoid of normal vision by exposure to prism lenses that optically rotated their perceived world around the line of sight by 180 degrees. Adaptation to such prisms involved sustained and vigorous practice of the ability to redirect the unadapted efferent motor command; because prior to all visually guided movements, the to-be-executed efferent command was based on incorrect (prismatically reversed) spatial information. The time course of this sort of adaptation was systematically explored in Cooper-Shepard mental rotation (MR) tests and in naturalistic motor-tasks for the purpose of investigating whether mental rotations of the direction of the intended movement share common aspects with the process of MR. A control group ( n=7) intermittently exposed to the distorted spatial organization of the central visual field was studied in parallel. The main results were as follows: (a) the MR reaction times (RTs) day 1 with prisms appeared to be very similar to the normal RTs (day 1, no-prisms) with the one exception that subjects now responded within a prism (rotated) frame of spatial reference rather than within the environmentally upright. The visuomotor performance became grossly irregular and dysmetric. (b) The majority of the visuomotor adaptation functions began to level off on the 3rd day. (c) The increases in natural motor proficiency were accompanied by a systematic and noticeable decrease in magnitude of the MR Y-intercept obtained from the linear regression line calculated between each subject's RT and the various stimulus angles. MR slopes were stable through days 1-7 for both the experimental and control group. An increased correlation between rotational stimulus angle and RT suggested that the MR function also became progressively more tightly coupled to the stimulus angles. (d) Postadaptation measures of performance indicated the occurrence of selective and minimal adaptation in the natural motor tasks only. It is suggested that these results reflect an improved attentional (strategic) ability to replace incorrect (error producing) control signals with correct (error reducing) control signals. As a result, perceptual-motor start-up processes directly related to spatial coding and to the planning, initiation and correction of the intended direction of motor-or-mental movement improved while the subprocess ("stage") concerned with transformations of such movements remained unchanged. Visuomotor adaptation to inverting prisms engages, and thereby stimulates, a cortical system also invoked in the preparatory process of MR.     

Dangerous liaisons: complement,coagulation, and kallikrein/kinin cross-talk act as a linchpin in the events leading to thromboinflammation

Innate immunity is fundamental to our defense against microorganisms. Physiologically, the intravascular innate immune system acts as a purging system that identifies and removes foreign substances leading to thromboinflammatory responses, tissue remodeling, and repair. It is also a key contributor to the adverse effects observed in many diseases and therapies involving biomaterials and therapeutic cells/organs. The intravascular innate immune system consists of the cascade systems of the blood (the complement, contact, coagulation, and fibrinolytic systems), the blood cells (polymorphonuclear cells, monocytes, platelets), and the endothelial cell lining of the vessels. Activation of the intravascular innate immune system in vivo leads to thromboinflammation that can be activated by several of the system's pathways and that initiates repair after tissue damage and leads to adverse reactions in several disorders and treatment modalities. In this review, we summarize the current knowledge in the field and discuss the obstacles that exist in order to study the cross-talk between the components of the intravascular innate immune system. These include the use of purified in vitro systems, animal models and various types of anticoagulants. In order to avoid some of these obstacles we have developed specialized human whole blood models that allow investigation of the cross-talk between the various cascade systems and the blood cells. We in particular stress that platelets are involved in these interactions and that the lectin pathway of the complement system is an emerging part of innate immunity that interacts with the contact/coagulation system. Understanding the resulting thromboinflammation will allow development of new therapeutic modalities.     

Heritability and the Equal Environments Assumption: Evidence from Multiple Samples of Misclassified Twins

Classically derived estimates of heritability from twin models have been plagued by the possibility of genetic-environmental covariance. Survey questions that attempt to measure directly the extent to which more genetically similar kin (such as monozygotic twins) also share more similar environmental conditions represent poor attempts to gauge a complex underlying phenomenon of GE-covariance. The present study exploits a natural experiment to address this issue: Self-misperception of twin zygosity in the National Longitudinal Survey of Adolescent Health (Add Health). Such twins were reared under one “environmental regime of similarity” while genetically belonging to another group, reversing the typical GE-covariance and allowing bounded estimates of heritability for a range of outcomes. In addition, we examine twins who were initially misclassified by survey assignment—a stricter standard—in three datasets: Add Health, the Minnesota Twin Family Study and the Child and Adolescent Twin Study in Sweden. Results are similar across approaches and datasets and largely support the validity of the equal environments assumption.     

Obesity prevention and personal responsibility: the case of front-of-pack food labelling in Australia

Background  

In Australia, the food industry and public health groups are locked in serious struggle for regulatory influence over the terms of front-of-pack food labelling. Clear, unambiguous labelling of the nutritional content of pre-packaged foods and of standardized food items sold in chain restaurants is consistent with the prevailing philosophy of 'personal responsibility'. An interpretive, front-of-pack labelling scheme has the capacity to encourage healthier patterns of eating, and to be a catalyst for improvements in the nutritional quality of food products through re-formulation. On the other hand, the strength of opposition of the Australian Food and Grocery Council to 'Traffic Light Labelling', and its efforts to promote a non-interpretive, voluntary scheme, invite the interpretation that the food industry is resistant to any reforms that could destabilise current (unhealthy) purchasing patterns and the revenues they represent.     

Associations between androgen and Vitamin D receptor microsatellites and postmenopausal breast cancer.

We investigated the association between polymorphism in the androgen receptor (AR) and vitamin D receptor (VDR) genes and breast cancer risk in a large population-based case-control study of genetically homogenous Swedish women. We successfully determined both AR CAG(n) and VDR A(n) genotype in 1,502 women with invasive breast cancer and in 1,510 control women. We did not find any associations between AR or VDR microsatellite lengths and breast cancer when we used a priori determined cutoffs (/=22 repeats for AR and /=19 for VDR) to define long and short alleles. There was statistically significant interaction between VDR genotype and parity, such that women with two short alleles had a halved risk for breast cancer, irrespective of parity, compared with nulliparous women with two long alleles. Homozygosity for the long VDR allele was associated with a more advanced clinical stage at diagnosis. In exploratory analyses, we determined cutoffs based on visual inspection of distributions of allele lengths among cases and controls and found that women carrying two alleles with <20 AR CAG(n) repeats had an increased risk for breast cancer, odds ratio of 1.67 (95% confidence interval, 1.17-2.38), compared with those with two alleles with >/=20 repeats. Women carrying two VDR alleles with <21 A(n) were also at an increased risk, odds ratio of 1.26 (95% confidence interval, 1.04-1.51). Our data do not support major roles for AR or VDR polymorphism as breast cancer risk factors. However, we did find an interaction between VDR genotype and parity that remains to be corroborated.     

Effect of proprioceptor stimulation on postural stability in patients with peripheral or central vestibular lesion

Body sway in upright stance at rest and after inducing proprioceptor stimulation, elicited by vibration applied to the calf or neck muscles, was studied in 11 patients with peripheral lesion and in 17 patients with central vestibular lesion. The responses were compared with those of 20 normal subjects. Vibratory stimulus was applied at five different frequencies, ranging from 32 to 150 Hz, and at a constant amplitude of 2.1 mm. Postural stability was measured with a force platform in terms of average deviation of body position (ADBP) analyzed in relation to the individual maximum support distance in the anterio-posterior direction. In patients with peripheral vestibular lesion ADBP was moderately increased, compared to normal subjects, when the calf muscles were exposed to vibration under eyes closed conditions (i.e. no visual information available); stimulation of neck muscles both under eyes open and eyes closed conditions and stimulation of calf muscles with open eyes produced an ADBP of the same magnitude as in controls. In patients with central vestibular lesion, proprioceptor stimulation of calf and neck muscles caused increased ADBP whether with eyes open or closed. The ADBP induced by stimulation of neck muscles was significantly greater in patients with a central lesion than in those with a peripheral vestibular lesion. The results indicate that patients with peripheral lesion differ from those with central vestibular lesion in their reaction to proprioceptor stimulus; and that in patients with central vestibular lesion proprioceptor stimulation of the neck muscles produces disproportionately powerful cervico-collic reflexes.     

Postural compensation in children with congenital or early acquired bilateral vestibular loss

The relative roles of different receptor systems in compensating for vestibular loss were studied in 18 children (12 to 16 years of age) with congenital or early acquired bilateral vestibular loss (BVL) and impaired hearing, and compared to that in 33 normal children (9 to 16 years of age). Postural stability was studied with posturography evaluating the velocity of anterior-posterior body sway. With closed eyes the increase of sway velocity did not differ between the groups, either with or without simultaneous proprioceptive disturbances when the subjects were standing on a bare surface. Body sway velocities were found to increase more in subjects with a BVL than in normal children when subjects were standing on foam rubber. When proprioceptive cues were disturbed, body sway velocities increased in both groups, but significantly more in children with BVL when subjects stood on foam rubber. This occurred both with and without open eyes.     

The relationship between caloric response, oculomotor dysfunction and size of cerebello-pontine angle tumours

Seventy-nine consecutive patients, 70 patients with acoustic neurinomas (ACN) and 9 patients with cerebello-pontine angle tumours of other etiology (o-CPA) were analysed with respect to tumour size and the result of the caloric and oculomotor tests. The size of the tumour was judged by the largest diameter calculated from the computerized tomography. A highly significant linear relationship between tumour size and caloric side difference was found for the group of ACN patients who had normal oculomotor function. A unilateral loss of caloric response on the tumour side was found in 75% of the ACN patients with tumours larger than 20 mm, but not in any of the ACN patients with tumours smaller than 10 mm. Oculomotor dysfunction (OMD), defined as disturbed pursuit eye movements and/or gaze nystagmus was frequently found in the patients with ACN larger than 20 mm (77%), and was present in all but one of the o-CPA patients. However, the caloric response on the tumour side was significantly more impaired in the group of ACN patients with OMD than in the o-CPA group. A combined view of the oculomotor and caloric test results offers a possibility to obtain a rough estimate of tumour size as well as to distinguish ACNs from other types of tumours in the cerebello-pontine angle.