BACKGROUND

A field of study which has attracted much recent interest is the ability of mononuclear cells and neutrophils to interact with histamine releasing cells by production of specific histamine releasing factors (HRFs). However, almost all of these studies have been performed on basophils rather than human mast cells.

OBJECTIVE

We have investigated the effects of lyophilized fractions of HRF preparations on histamine release from human skin and lung mast cells.

METHODS

Lyophilized fractions of HRF preparations include crude supernatant from mononuclear cell/platelet (crude), void peak from anion exchange chromatography column (void), second peak from anion exchange chromatography (peak 2), neutrophil-activating peptide-2 (NAP-2), which was purified from void peak at molecular weight of 8-12 kDa, and monocyte chemotactic-activating factor (MCAF). Mast cells were enzymatically dispersed.

RESULTS

Crude (24.2 micrograms/mL-2.42 mg/mL), void (5.4 micrograms/mL-0.54 mg/mL), peak 2 (3.5 micrograms/mL-0.35 mg/mL), and NAP-2 (1-20 micrograms/mL) failed to release histamine from lung mast cells. In skin mast cells, only higher concentrations of crude and void caused minimal release of histamine. MCAF up to micromolar concentrations failed to have an effect on mast cells from either source. However, these HRFs induced histamine release from human basophils. We also explored whether HRFs and stem cell factor could act as either priming agents for each other or for anti-IgE. The response of skin mast cells to all these preparations was not enhanced by preincubation in stem cell factor at 1 ng/mL, nor did the HRFs and MCAF enhance the response of skin mast cells to anti-IgE.

CONCLUSIONS

These results suggest that these HRFs have no significant effect on dispersed human cutaneous and lung mast cells.

We demonstrated the production of a histamine releasing factor (HRF) by 24-h cultures of guinea pig spleen cells which were stimulated or not with specific antigen (ovalbumin, OA) or mitogen (phytohemagglutinins or concanavalin A). HRF induced the release of histamine from homologous mesenteric mast cells in a dose-dependent fashion. The HRF-induced histamine release was not high compared to the release induced by calcium ionophore A23187, but higher than that induced by compound 48/80, polymyxin B and con canavalin A. The mast cells from sensitized guinea pigs released histamine when challenged with OA. We found that HRF-induced histamine release was additive to that induced by antigen, when both agents were added simultaneously to sensitized mast cells. The phenomenon was most significant when a suboptimal dose of antigen was used. Moreover, we did not observe any differences in the magnitude of HRF-induced histamine release between the mast cells from nonsensitized and sensitized guinea pigs. The time course of histamine release induced by HRF was significantly slower than that with specific antigen (10 min and 45 sec, respectively). Our results may suggest that HRF acts on mast cells through a different not immunological mechanism.

We investigated the relation between IgE reactive with histamine-releasing factor (HRF) and clinical status in patients with asthma or rhinitis. Sera were used to passively sensitize purified, lactic-acid treated basophils. IgE-independent HRF due to chemokines was removed from mononuclear cell supernatants with heparin-Sepharose. IgE-dependent HRF was determined by measuring the increase in histamine release between 1 min and 60 min, which was designated delta HRF. HRF-reactive IgE was demonstrable in nine of 18 patients with allergic asthma, three of 19 patients with nonallergic asthma, five of 17 patients with allergic rhinitis, and none of 19 control patients. The presence of HRF-reactive IgE was associated with: (1) IgE to inhalant allergens; 40% of radioallergosorbent test (RAST)-positive individuals versus 8% of RAST-negative individuals were positive (OR = 7.8, p < 0.005); (2) bronchial sensitivity to histamine in all asthmatic patients (geometric mean PC20: 1.50 versus 0.51 mg/ml; p < 0.005); and (3) bronchial sensitivity to histamine in allergic asthmatic patients (geometric mean PC20: 1.27 versus 0.37 mg/ml, p < 0.02). These findings support the hypothesis that IgE-dependent HRF might contribute to the chronic allergic reaction.

A pore-forming protein (PFP/perforin/cytolysin), stored in the cytoplasmic granules of cytolytic lymphocytes, lyses a variety of target cells but not the cytolytic lymphocytes. In the complement (C) system, a C8-binding protein (C8bp) or homologous restriction factor (HRF) has been described that protects cells against lysis mediated by homologous C. C8bp/HRF is known to bind to C8 and C9 and has also been suggested to protect lymphocytes against perforin-mediated lysis. Here, using an anti-idiotypic antibody approach, several polyclonal antisera were raised against IgGs that are specific for mouse perforin, and human C8 and C9. These anti-idiotypic antisera were shown to react against an overlapping epitope(s) on C8bp/HRF as indicated by the following evidence: (i) all three types of antisera reacted against partially purified C8bp/HRF and against a 65 kDa protein band in cell lysates; reactivity was only observed against disulfide-reduced antigens; (ii) the three antibodies react with a protein band in normal erythrocytes (E) but not with type III E of patients with paroxysmal nocturnal hemoglobinuria or with a mutant B lymphoblastoid cell line, both of which cell types are known to be deficient in C8bp/HRF; and (iii) the three antibodies compete with each other for binding to C8bp/HRF. Type III E and the C8bp/HRF-deficient mutant lymphoblastoid cell line, however, are as susceptible to perforin-mediated lysis as type I E and wild-type lymphoblastoid cell line, respectively, indicating that C8bp/HRF does not play a role in protecting cells against perforin-mediated lysis. These paradoxical findings suggest that perforin may share with C8 and C9 the same domain(s) that bind to C8bp/HRF and yet, unlike C8 and C9, perforin is not inactivated by this type of putative interaction. Since C8 and C9 are now readily available, the anti-idiotypic approach described here provides a convenient protocol for production of antisera specific for C8bp/HRF.

The aim of the study was to determine whether the Heidelberg Retinal Flowmeter (HRF), a confocal scanning laser Doppler flowmeter, can measure choroidal blood flow in pig eyes.An HRF was used to obtain flow maps from in vitro pig eyes under a range of perfusion flow rates (0 - 500 microL/min) under conditions in which only the choroid was perfused. In some cases choroidal blood flow was also measured simultaneously using a conventional fiberoptic based Laser Doppler Perfusion Monitor (LDPM) which used the same laser wavelength (780 nm). The relationship between perfusion flow, HRF measured flow and LDPM measured flow was determined. HRF flow maps were also obtained in vivo as a function of focal plane setting through the retina and choroid.Across the range of perfusion flow rates through the isolated eyes there was a poor correlation with HRF measured choroidal flow and perfusion flow. In contrast, there was a strong linear relationship between perfusion flow and LDPM measured blood flow. Both in vitro and in vivo, no choroidal vessels could be visualised in the HRF flow maps, even when the focal plane was in the choroid.The HRF is unable to measure choroidal blood flow in pig eyes. This is not due to an inability of the 780 nm laser to penetrate into the choroid or due to red blood cell velocities in the choroid being higher than the measurement range of the instrument.

OBJECTIVE

As retinal vessels in color fundus images are thin and elongated structures, standard pairwise based random fields, which always suffer the "shrinking bias" problem, are not competent for such segmentation task. Recently, a dense conditional random field (CRF) model has been successfully used in retinal vessel segmentation. Its corresponding energy function is formulated as a linear combination of several unary features and a pairwise term. However, the hand-crafted unary features can be suboptimal in terms of linear models. Here we propose to learn discriminative unary features and enhance thin vessels for pairwise potentials to further improve the segmentation performance.

METHODS

Our proposed method comprises four main steps: firstly, image preprocessing is applied to eliminate the strong edges around the field of view (FOV) and normalize the luminosity and contrast inside FOV; secondly, a convolutional neural network (CNN) is properly trained to generate discriminative features for linear models; thirdly, a combo of filters are applied to enhance thin vessels, reducing the intensity difference between thin and wide vessels; fourthly, by taking the discriminative features for unary potentials and the thin-vessel enhanced image for pairwise potentials, we adopt the dense CRF model to achieve the final retinal vessel segmentation. The segmentation performance is evaluated on four public datasets (i.e. DRIVE, STARE, CHASEDB1 and HRF).

RESULTS

Experimental results show that our proposed method improves the performance of the dense CRF model and outperforms other methods when evaluated in terms of F1-score, Matthews correlation coefficient (MCC) and G-mean, three effective metrics for the evaluation of imbalanced binary classification. Specifically, the F1-score, MCC and G-mean are 0.7942, 0.7656, 0.8835 for the DRIVE dataset respectively; 0.8017, 0.7830, 0.8859 for STARE respectively; 0.7644, 0.7398, 0.8579 for CHASEDB1 respectively; and 0.7627, 0.7402, 0.8812 for HRF respectively.

CONCLUSIONS

The discriminative features learned in CNNs are more effective than hand-crafted ones. Our proposed method performs well in retinal vessel segmentation. The architecture of our method is trainable and can be integrated into computer-aided diagnostic (CAD) systems in the future.

The problem of model falsification or model invalidation appears in several areas where we are interested in distinguishing among an eligible set of dynamic systems. In the context of fMRI studies of brain activity, modeling the haemodynamic response function (HRF) is a critical step. The estimation of the dynamic system describing a biophysical model of the HRF may leave much uncertainty on the exact values of the parameters. Moreover, the high noise levels in the data may hinder the model identification task. Therefore, this paper proposes a systematic tool to address the problem of the distinguishability among a set of physiologically plausible HRF models. The concept of absolutely input distinguishable systems is introduced and applied to the HRF model, by exploiting the structure of the underlying nonlinear dynamic system. A strategy to model uncertainty in the input time delay and magnitude is developed and its impact on the distinguishability of two physiologically plausible HRF models is determined, in terms of the maximum noise amplitude above which it is not possible to guarantee the falsification of one model in relation to the other. Finally, a methodology is proposed for the choice of the input sequence, or experimental paradigm, that should be used in order to maximize the distinguishability of the HRF models under investigation. The proposed approach may be used to assess the performance of HRF model identification techniques from fMRI data.

This study investigated the kinematics and kinetics of the carpus during the stance phase. Five Standardbred horses trotted on a treadmill at 8.9 m/s. The kinematics of the horses were filmed and hoof reaction forces (HRF) were recorded. The carpus was overextended throughout most of the stance. There were 2 periods of overextension, a more rapid period in the beginning of the stance and second directly following the first period. Maximal overextension occurred slightly before the second minimum of the braking horizontal HRF. The metacarpal and antebrachial segments rotated counter-clockwise for most of the stance. The angular velocities of these segments attained absolute and local extremes that were concurrent. The absolute maxima in the longitudinal and the transversal acceleration of the proximal metacarpus were coincident in time with the minimal horizontal braking force. The moment of force about the carpus was extending throughout most of the stance, with a short period of flexion near the end of the stance. The results show that horizontal braking of the hoof results in impulsive loading followed by maximal overextension of the carpus. The second phase of overextension is suggested to be related to the tension or active roll of the flexor tendons of the distal limb. The carpus is, therefore, subjected to rapid and high loading in both the longitudinal and transversal directions, as well as large moments and forces.

Brain state decoding or "mind reading" via multivoxel pattern analysis (MVPA) has become a popular focus of functional magnetic resonance imaging (fMRI) studies. In brain decoding, stimulus presentation rate is increased as fast as possible to collect many training samples and obtain an effective and reliable classifier or computational model. However, for extremely rapid event-related experiments, the blood-oxygen-level-dependent (BOLD) signals evoked by adjacent trials are heavily overlapped in the time domain. Thus, identifying trial-specific BOLD responses is difficult. In addition, voxel-specific hemodynamic response function (HRF), which is useful in MVPA, should be used in estimation to decrease the loss of weak information across voxels and obtain fine-grained spatial information. Regularization methods have been widely used to increase the efficiency of HRF estimates. In this study, we propose a regularization framework called mixed L2 norm regularization. This framework involves Tikhonov regularization and an additional L2 norm regularization term to calculate reliable HRF estimates. This technique improves the accuracy of HRF estimates and significantly increases the classification accuracy of the brain decoding task when applied to a rapid event-related four-category object classification experiment. At last, some essential issues such as the impact of low-frequency fluctuation (LFF) and the influence of smoothing are discussed for rapid event-related experiments.

OBJECTIVE

The functional independence of elderly populations deteriorates with age. Several tests of physical performance have been developed for screening elderly persons who are at risk of losing their functional independence. The purpose of the present study was to investigate whether several components of health-related fitness (HRF) are valid in predicting the occurrence of self-reported mobility difficulties (MD) among high-functioning older adults.

METHODS

Subjects were community-dwelling men and women, born 1917-1941, who participated in the assessment of HRF [6.1-m (20-ft) walk, one-leg stand, backwards walk, trunk side-bending, dynamic back extension, one-leg squat, 1-km walk] and who were free of MD in 1996 (no difficulties in walking 2- km, n=788; no difficulties in climbing stairs, n=647). Postal questionnaires were used to assess the prevalence of MD in 1996 and the occurrence of new MD in 2002. Logistic regression analysis was used as the statistical method.

RESULTS

Both inability to perform the backwards walk and a poorer result in it were associated with risk of walking difficulties in the logistic model, with all the statistically significant single test items included. Results of 1-km walk time and one-leg squat strength test were also associated with risk, although the squat was statistically significant only in two older birth cohorts. Regarding stair-climbing difficulties, poorer results in the 1-km walk, dynamic back extension and one-leg squat tests were associated with increased risk of MD.

CONCLUSIONS

The backwards walk, one-leg squat, dynamic back extension and 1-km walk tests were the best predictors of MD. These tests are recommended for use in screening high-functioning older people at risk of MD, as well as to target physical activity counseling to those components of HRF that are important for functional independence.

Non-invasive ventilation (NIV) with bilevel positive airway pressure is a non-invasive technique, which refers to the provision of ventilatory support through the patient's upper airway using a mask or similar device. This technique is successful in correcting hypoventilation. It has become widely accepted as the standard treatment for patients with hypercapnic respiratory failure (HRF). Since the 1980s, NIV has been used in intensive care units and, after initial anecdotal reports and larger series, a number of randomized trials have been conducted. Data from these trials have shown that NIV is a valuable treatment for HRF. This review aims to explore the principal areas in which NIV can be useful, focusing particularly on patients with acute HRF (AHRF). We will update the evidence base with the goal of supporting clinical practice. We provide a practical description of the main indications for NIV in AHRF and identify the group of patients with hypercapnic failure who will benefit most from the application of NIV.

Toxicology feeding studies of mineral oil hydrocarbons (MOHs), within the carbon number range C22-C28, results in species-specific epithelioid granulomas in the liver of F-344 rats but not in other rat strains, or species. While MOH has been detected, and some pathological effects have been shown to occur in other organs/tissues of F-344 rats and other rat strains/species, it is generally accepted that the effect of toxicological concern is species-specific inflammatory liver granuloma. As oil retention and other MOH-related nontoxic pathological changes in the liver are observed in humans, some have hypothesized that the potential for oil accumulation over a lifetime, through dietary sources, may predispose humans to similar liver effects as observed in F-344 rats. To address this concern, a mode of action/human relevance framework (MoA/HRF) analysis for MOH-induced epithelioid granuloma in the F-344 rat model was developed. The key events for the development of liver epithelioid granulomas were identified as increased MOH intestinal absorption, preferential tissue retention and ultimately formation of necrotic granulomas encased by infiltrating inflammatory lymphocytes. The hypothesized MoA was evaluated using the modified Bradford Hill considerations for causality and was considered to be established in the F-344 rodent model. However, key strain/species differences in the rate of intestinal absorption, tissue retention of MOH and inflammatory response to MOH in the liver were identified. Overall, the F-344 rat MoA was not considered to be relevant to humans, consistent with data showing no evidence for the formation of epithelioid granulomas with humans even in cases of massive ingestion of MOHs.

A cDNA that codes for the polypeptide hormone precursor proopiomelanocortin (POMC) was cloned and sequenced from a gar (Lepisosteus osseus) pituitary cDNA library. The gar POMC cDNA is 1237 bp and contains a 780-bp open reading frame. The deduced amino acid sequence for gar POMC is 259 amino acids in length. The general organization of gar POMC is very similar to that of other gnathostome POMC sequences. The beta-endorphin sequence had 91% sequence identity with sockeye A beta-endorphin and 71% sequence identity with Xenopus laevis beta-endorphin. Three melanocyte-stimulating hormone (MSH) core sequences [HFR(W)] were detected. The gar alpha-MSH sequence was identical to the alpha-MSH sequence in rat POMC. The gar beta-MSH sequence had 77% sequence identity with salmonid forms of beta-MSH and 53% sequence identity with tetrapod forms of beta-MSH. The gamma-MSH region of gar POMC only had 26% primary sequence identity with tetrapod gamma-MSH sequences. Gar gamma-MSH had an incomplete MSH core sequence (HRF), an apparent internal deletion of five amino acids, and lacked flanking paired basic amino acids essential for proteolytic cleavage. The apparent degenerate nature of gar gamma-MSH is discussed in light of the absence of this sequence in salmonid fish.

In allergic disease, mast cell activation is conventionally triggered by allergen-mediated cross-linking of receptor-bound IgE on the cell surface. In addition to its diverse range of intracellular roles in apoptosis, cell proliferation and cancer, Histamine-Releasing Factor (HRF) also activates mast cells and basophils. A subset of IgE antibodies bind HRF through their Fab regions, and two IgE binding sites on HRF have been mapped. HRF can form dimers, and a disulphide-linked dimer is critical for activity. The current model for the activity of HRF in mast cell activation involves cross-linking of receptor-bound IgE by dimeric HRF, mediated by HRF/Fab interactions. HRF crystal and solution structures have provided little insight into either the formation of disulphide-linked HRF dimers or the ability of HRF to activate mast cells. We report the first crystal structure of murine HRF (mHRF) to 4.0Å resolution, revealing a conserved fold. We also solved the structure of human HRF (hHRF) in two new crystal forms, one at the highest resolution (1.4Å) yet reported. The high resolution hHRF structure reveals a disulphide-linked dimer, in which the two molecules are closely associated, and provides a model for the role of both human and murine HRF in mast cell activation.

Since basophils appear to play a fundamental role in the maintenance of allergic inflammation, the factors involved in basophil activation have become a focus of investigation in many laboratories. From this interest, the field of histamine-releasing factors (HRFs) has evolved. Our laboratory reported that a factor was present in late-phase skin blister fluids that caused basophil histamine release. It was hypothesized that basophil degranulation in these donors was mediated by the interaction of these HRFs with a certain kind of IgE, and upon experimentation a functional heterogeneity of the IgE molecule was uncovered. We designated the IgE from HRF responders as IgE+; the remaining IgE molecules designated were IgE-. Initially, it was thought that HrHRF might exert its activity by directly interacting with IgE+; however, a number of recent experiments have questioned this hypothesis. Observations suggest, in part, that HrHRF mediates biological activities on inflammatory cells by binding to a specific receptor rather than to the IgE molecule and/or the FcepsilonRI. In parallel with the search for an HrHRF receptor, a number of experiments have been performed to determine whether mediator release by this protein proceeds via a signal transduction pathway other than the one triggered by classic IgE-dependent stimuli such as anti-IgE antibody or antigen. Although further characterization is ongoing, the evidence thus far is consistent with the concept that HRF may be an important regulator of the cellular inflammation involved in the pathophysiology of allergy.

The Autonomic Information Flow (AIF) represents the complex communication within the autonomic nervous system (ANS). It can be assessed by the mutual information function (MIF) of heart rate fluctuations (HRF). AIF represented by HRF is based on several interacting physiological mechanisms operating at different time scales. Therefore one prominent time scale for HRF complexity analysis is not given a priori. MIF reflects the information flow at different time scales. This allows a more specific characterization of the complex communication leading to dynamic stability (homeostasis) of the cardiovascular-respiratory system. Three clinically relevant examples of autonomic communication, namely AIF of normals during awake state and deep sleep, and of patients with multiple organ dysfunction syndrome (MODS) were investigated. In these states MIF clearly represents and distinguishes the different communication in dependence on the time scale of information transfer. The results confirm our systems-theoretical concept that AIF, represented by MIF, characterizes the complex communication of the ANS at different time scales.

The simultaneous acquisition of electroencephalogram (EEG) and functional MRI (fMRI) signals is potentially advantageous because of the superior resolution that is achieved in both the temporal and spatial domains, respectively. However, ballistocardiographic artifacts along with ocular artifacts are a major obstacle for the detection of the EEG signatures of interest. Since the sources corresponding to these artifacts are independent from those producing the EEG signatures, we applied the Infomax-based independent component analysis (ICA) technique to separate the EEG signatures from the artifacts. The isolated EEG signatures were further utilized to model the canonical hemodynamic response functions (HRFs). Subsequently, the brain areas from which these EEG signatures originated were identified as locales of activation patterns from the analysis of fMRI data. Upon the identification and subsequent evaluation of brain areas generating interictal epileptic discharge (IED) spikes from an epileptic subject, the presented method was successfully applied to detect the theta and alpha rhythms that are sleep onset-related EEG signatures along with the subsequent neural circuitries from a sleep-deprived volunteer. These results suggest that the ICA technique may be useful for the preprocessing of simultaneous EEG-fMRI acquisitions, especially when a reference paradigm is unavailable.

The release of mediators from mast cells and basophils represents the central event in the development of immediate hypersensitivity reactions with release of substances such as histamine, Leukotrienes C4/D4, Platelet Activating Factor (PAF), and Prostaglandin D2. Cytokines such as IL-1, TNF alpha, and IL-4 may also be secreted. Histamine Releasing Factors (HRF) are cytokine-like molecules that interact with basophils and/or mast cells to cause cell activation and secretion of mediators. Histamine release is the best characterized of these and has been used as the assay for HRFs, but a wide variety of inflammatory mediators can potentially be secreted. We believe this type of cell to cell communication to be important in tissue inflammation, in which infiltrating cells may produce HRFs in proximity to infiltrating basophils and/or mast cells and cause them to degranulate. Such a reaction appears to be independent of IgE antibody, may no longer require the presence of any inciting antigen, and appears to be pertinent to the allergic late phase reaction as it occurs in the nose, lungs, and skin. It is thought that an ongoing antigenic stimulus, as seen in seasonal or perennial allergic rhinitis and asthma, or in certain types of urticaria, or in atopic dermatitis, leads to a perpetuating inflammatory reaction that persists for many weeks or months. A chronic inflammatory reaction of this sort appears to be required for an allergic reaction (IgE mediated) to manifest as an allergic disease. The relationship of HRF to the chemokine group of cytokine-like molecules and the importance of HRF in the pathogenesis of bronchial hyperreactivity and asthma has been reviewed in this paper.

The 65 kDa C8-binding protein or homologous restriction factor (C8bp/HRF) protects cells from complement (C)-mediated lysis by binding to C8 and abrogating lytic channel formation. Human C8bp/HRF is shown here to be immunologically related to human C8 and C9 and to murine lymphocyte poreforming protein (PFP, perforin). Polyclonal antibodies raised against purified C8, C9 and perforin react with C8bp/HRF. The antigenic epitopes shared by these four proteins are limited to cysteine-rich or disultide bridge-masked domains. Only complement proteins or perforin that have been disulfide-reduced elicit the production of cross-reactive antibodies when used as immunogens. Analogously, only C8bp/HRF that has been disulfide-reduced reacts with these antibodies. These results suggest that C8bp/HRF may belong to the complement/perforin supergene family. The function of homologous domains shared by these four proteins remains to be elucidated.

It has been suggested that differential histamine-releasing activity of an IgE-dependent histamine-releasing factor (HRF), which has recently been cloned, is related to carbohydrate difference in the IgE molecule. Lectins are able to recognize specific glycoforms and might therefore be useful in characterizing the proposed heterogeneity of IgE molecules. As one test of this hypothesis, we examined the histamine release potency of several well-characterized lectins on basophils passively sensitized with serum containing IgE molecules that support HRF-induced histamine release (IgE+) or serum that does not support release by this stimulus (IgE-). Histamine release was induced by challenging basophils with different concentrations of concanavalin A, Lens culinaris (LcH), and Pisum sativum (PSA). Dose-response curves revealed that LcH caused 30% histamine release at 2 micrograms/ml with IgE+ sensitized cells, whereas the same release with IgE- cells required sixfold higher concentrations. Similar values for PSA showed a sevenfold difference. With concanavalin A, the selectivity was reversed in that it was fourfold more active on IgE- -sensitized cells. Another pair of sera (IgE+ vs IgE-) revealed the same result for concanavalin A, but no difference occurred in LcH and PSA induced release between the IgE+ - and IgE- -sensitized cells. These contrasting findings with different pairs of IgE+ -and IgE- -containing sera indicated that the lectins LcH and PSA are not able to discriminate between IgE+ -and IgE- -sensitized cells as does HRF and therefore cannot be used to further define the proposed heterogeneity of IgE. this conclusion was supported by experiments in which basophil preparations from donors possessing natural sensitivity or insensitivity to HRF (having IgE+ or IgE- on their surface) were examined fro their response to these lectins. No difference was found in the sensitivity of the cells to challenge with LcH or PSA, and the response to concanavalin A was the opposite of that found when passively sensitized cells were used (30% histamine release at 0.9 vs 3.5 micrograms/ml for IgE+ vs IgE- donors, respectively). We conclude that oligosaccharide-specific lectins do not differentiate between HRF-reactive and HRF-nonreactive IgE molecules on basophils.

Protein kinase C (PKC) and mitogen-activated protein (MAP) kinase activation appear important in conferring hypertrophy in vitro. However, the response of PKC and MAP kinase to stimuli known to induce hypertrophy in vivo has not been determined. We recently demonstrated that pressure-overload hypertrophy induced a transiently transfected gene driven by an hypertrophy responsive enhancer (HRE) through a marked increase in binding activity of its interacting nuclear factor (HRF). These data suggested that the HRE/HRF could serve as a target for evaluating the signal transduction events responsible for hypertrophy in vivo. Accordingly, we characterized MAP kinase and PKC isoform activation, injected HRE driven reporter gene expression, and HRF binding activity in rat hearts subjected to ascending aortic clipping or sham operation in the presence of the angiotensin-converting enzyme (ACE) inhibitor fosinopril, hydralazine, or no treatment. Analyses showed that PKC-epsilon and MAP kinase were acutely activated following ascending aortic ligature and that fosinopril significantly inhibited but did not completely abrogate PKC-epsilon and MAP kinase activation. However, fosinopril completely prevented pressure overload-mediated induction of HRE containing constructs and obviated increased HRF binding activity. These results suggest a direct relationship between ACE activity and HRE/HRF-mediated gene activation and imply that PKC-epsilon and MAP kinase may be involved in transducing this signal.

Recurrent fevers are defined as multiple stereotypical febrile episodes separated by spontaneous symptom-free intervals and occurring for months and years. Hereditary recurrent fevers are rare prototype Mendelian diseases due to inherited mutations in genes encoding partners of the innate immunity. Recurrent episodes of fever plus acute features of inflammation starting during childhood with family history are the main clues for suspecting HRF. Their common associated complication is AA amyloidosis. The diagnosis is made on clinical grounds but the genetic diagnosis may contribute in most cases of monogenic hereditary recurrent fevers. Recurrent fevers must be distinguished from intermittent fevers, mostly infectious, characterized by variation in associated symptoms from episode-to-episode and without periodicity.