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1.
Lymphology ; 56(2): 61-71, 2023.
Article in English | MEDLINE | ID: mdl-38621384

ABSTRACT

Primary lymphedema of the foot and toes could be sometimes misdiagnosed by lymphoscintigraphy as a whole lower limb lymphatic insufficiency (LLLI). This is caused by using standard lymphoscintigraphic protocol based on one interstitial injection of radiotracer applied into the first interdigital space followed by image analysis of lower limb lymphatic vessels and lymph nodes. Here, we show that a modification of the lymphoscintigraphic protocol and introduction of a second dose of radiotracer right above the inner ankle to the clinically healthy tissue can more accurately describe morphological abnormalities of the superficial lymphatic system at the lower limb and thereby refine the diagnosis of the LLLI. Fourteen patients with swelling of the foot and toes (16 lower limbs) were examined using standard lymphoscintigraphic protocol. Subsequently, modified lymphoscintigraphy was performed. While standard lymphoscintigraphy showed severe lymphatic insufficiency of the superficial lymphatic system in all 14 patients (in 16 lower limbs), including significantly reduced number of inguinal nodes, modified lymphoscintigraphy revealed almost normal morphology of superficial lymphatic vessels in 11 patients (in 13 lower limbs) throughout the entire lower limb proximal to the application site. In conclusion, using the modified lymphoscintigraphy protocol in patients with foot and toes primary lymphedema can refine diagnosis and follow-up medical management.


Subject(s)
Lymphatic Vessels , Lymphedema , Humans , Lymphoscintigraphy/methods , Lymphedema/diagnostic imaging , Lymphedema/etiology , Lymphatic System/diagnostic imaging , Lymphatic Vessels/diagnostic imaging , Lower Extremity/diagnostic imaging
2.
Acta Paediatr ; 108(8): 1419-1426, 2019 08.
Article in English | MEDLINE | ID: mdl-30817025

ABSTRACT

AIM: The current study determined survival, short-term neonatal morbidity and predictors for death or adverse outcome of very preterm infants in Austria. METHODS: This population-based cohort study included 5197 very preterm infants (53.3% boys) born between 2011 and 2016 recruited from the Austrian Preterm Outcome Registry. Main outcome measures were gestational age-related mortality and major short-term morbidities. RESULTS: Overall, survival rate of all live-born infants included was 91.6% and ranged from 47.1% and 73.4% among those born at 23 and 24 weeks of gestation to 84.9% and 88.2% among infants born at 25 and 26 weeks to more than 90.0% among those with a gestational age of 27 weeks or more. The overall prevalence of chronic lung disease, necrotising enterocolitis requiring surgery, intraventricular haemorrhage Grades 3-4, and retinopathy of prematurity Grades 3-5 was 10.0%, 2.1%, 5.5%, and 3.6%, respectively. Low gestational age, low birth weight, missing or incomplete course of antenatal steroids, male sex, and multiple births were significant risk predictors for death or adverse short-term outcome. CONCLUSION: In this national cohort study, overall survival rates were high and short-term morbidity rate was low.


Subject(s)
Infant Mortality , Infant, Extremely Premature , Infant, Premature, Diseases/epidemiology , Austria/epidemiology , Female , Gestational Age , Humans , Infant , Infant, Newborn , Male
3.
Physiol Behav ; 173: 144-155, 2017 05 01.
Article in English | MEDLINE | ID: mdl-28167147

ABSTRACT

Much evidence has suggested that early life adversity can have a lasting effect on behavior. The aim of this study was to explore the impact of prenatal exposure to stress on cognition in adult life and how it impacts chronic stress situations. In addition, we investigated the participation of glucocorticoids, neurotrophins and cytokines in prenatal stress effects. For this purpose, pregnant mice were placed in a cylindrical restraint tube for 2h daily during the last week of pregnancy. Control pregnant females were left undisturbed during their entire pregnancy period. Object-in-place task results showed that adult female mice exposed to prenatal stress exhibited an impairment in spatial memory. However, in the alternation test this memory deficit was only found in prenatally stressed mice submitted to chronic stress. This alteration occurred in parallel with a decrease in BDNF, an increase in glucocorticoid receptors and an alteration of Th1/Th2 in the hippocampus. Interestingly, these changes were observed in peripheral lymph nodes as well. However, none of the mentioned changes were observed in adult male mice. These results indicate that lymphoid cells could be good candidates as peripheral markers of susceptibility to behavioral alterations associated with prenatal exposure to stress.


Subject(s)
Cytokines/metabolism , Lymphocytes/metabolism , Memory Disorders/etiology , Nerve Growth Factors/metabolism , Prenatal Exposure Delayed Effects/physiopathology , Stress, Psychological/complications , Animals , Brain/metabolism , Corticosterone/metabolism , Cytokines/genetics , Exploratory Behavior , Female , Male , Maze Learning/physiology , Memory Disorders/pathology , Mice , Nerve Growth Factors/genetics , Pregnancy , Receptors, Steroid/genetics , Receptors, Steroid/metabolism , Recognition, Psychology/physiology
4.
Bratisl Lek Listy ; 117(10): 595-600, 2016.
Article in English | MEDLINE | ID: mdl-27826975

ABSTRACT

AIM: The aim of our study was to ascertain the efficacy of low-molecular-weight heparin (LMWH) in the treatment of patients with SVT. MATERIAL AND METHODS: A group of 336 outpatients with clinical diagnosis of SVT was evaluated in this prospective study. At the beginning of the study all patients were examined by clinical investigation, laboratory tests and duplex ultrasound investigation (examination). All patients included into the study were treated with LMWH. Clinical and ultrasound evaluation was carried out on days 10, 30 and 90 of the follow-up. This clinical study of SVT treatment with LMWH was organized in 18 outpatient departments in the Slovak Republic. RESULTS: After 10 days of treatment with LMWH in full therapeutic dosage, an improvement in the clinical symptoms was demonstrated in 93 % of patients; a complete resolution of clinical symptoms was demonstrated in 4 % of patients. On day 30 (after 10 days of LMWH treatment in full therapeutic dosage and further treatment in halved therapeutic dosage up to 20 days) a complete resolution of clinical symptoms was observed in 59 % (n = 189) of patients. Patients were further clinically evaluated on day 90, after two months of no anticoagulation treatment. The clinical evaluation revealed a complete resolution of symptoms in 88 % (n = 283) of patients and improvement in symptoms in 11.6 % (n = 34). Two patients developed pulmonary embolization; extension of SVT was seen in one patient and SVT recurrence in two patients. CONCLUSION: Superficial vein thrombosis can propagate into the deep veins with the risk of pulmonary embolism. The results indicate that current ambulatory treatment regimen using LMWH in the treatment of SVT is effective and safe (Fig. 4, Ref. 30).


Subject(s)
Heparin, Low-Molecular-Weight/therapeutic use , Leg/blood supply , Venous Thrombosis/drug therapy , Adult , Aged , Ambulatory Care , Female , Humans , Male , Middle Aged , Prospective Studies , Slovakia , Treatment Outcome , Young Adult
5.
Klin Padiatr ; 226(4): 211-5, 2014 Jul.
Article in English | MEDLINE | ID: mdl-25010126

ABSTRACT

BACKGROUND: Intravenous sildenafil treatment has recently shown promising results and good tolerability in the treatment of refractory pulmonary hypertension (PH) in term and near-term neonates, while comparable data in preterm infants are still lacking. However, for critically ill preterm infants suffering from PH refractory to conventional treatment, sildenafil may represent a last treatment resort. PATIENTS AND METHODS: We reviewed the records of 6 critically ill extremely preterm infants who had suffered from PH refractory to conventional treatment and had obtained intravenous sildenafil after careful consideration as ultima ratio treatment. AIM: To describe the responses to sildenafil in terms of hemodynamic and respiratory changes during treatment and outcome. RESULTS: 4/6 patients showed resolution of severe PH with full reversal of ductal shunt direction into pure left-to-right shunt within 82 ± 35 h after sildenafil start. Remarkably, 2/6 patients developed pulmonary hemorrhage at a time point when significant improvement of PH had already taken place, both of them survived. Overall 4/6 patients died, two deaths were related to treatment-refractory PH. CONCLUSION: Intravenous sildenafil treatment seems effective in improving severe PH and hemodynamic instability in extremely preterm infants with refractory PH. Pulmonary hemorrhage may represent a distinct adverse effect of sildenafil treatment in these patients, presumably due to sudden reversal of ductal shunt. Accordingly, sildenafil should be restricted to most severe and refractory cases in this population.


Subject(s)
Infant, Extremely Low Birth Weight , Infant, Premature, Diseases/drug therapy , Persistent Fetal Circulation Syndrome/drug therapy , Piperazines/administration & dosage , Sulfonamides/administration & dosage , Vasodilator Agents/administration & dosage , Female , Hemorrhage/chemically induced , Hemorrhage/diagnosis , Humans , Infant, Newborn , Infant, Premature, Diseases/diagnosis , Infusions, Intravenous , Intensive Care Units, Neonatal , Lung Diseases/chemically induced , Lung Diseases/diagnosis , Male , Persistent Fetal Circulation Syndrome/diagnosis , Piperazines/adverse effects , Pulmonary Wedge Pressure/drug effects , Purines/administration & dosage , Purines/adverse effects , Retrospective Studies , Sildenafil Citrate , Sulfonamides/adverse effects , Vasodilator Agents/adverse effects
6.
Osteoporos Int ; 25(3): 973-82, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24221453

ABSTRACT

SUMMARY: We investigated the association of postmenopausal vertebral deformities and fractures with bone parameters derived from distal extremities using MRI and pQCT. Distal extremity measures showed variable degrees of association with vertebral deformities and fractures, highlighting the systemic nature of postmenopausal bone loss. INTRODUCTION: Prevalent vertebral deformities and fractures are known to predict incident further fractures. However, the association of distal extremity measures and vertebral deformities in postmenopausal women has not been fully established. METHODS: This study involved 98 postmenopausal women (age range 60-88 years, mean 70 years) with DXA BMD T-scores at either the hip or spine in the range of -1.5 to -3.5. Wedge, biconcavity, and crush deformities were computed on the basis of spine MRI. Vertebral fractures were assessed using Eastell's criterion. Distal tibia and radius stiffness was computed using MRI-based finite element analysis. BMD at the distal extremities were obtained using pQCT. RESULTS: Several distal extremity MRI and pQCT measures showed negative association with vertebral deformity on the basis of single parameter correlation (r up to 0.67) and two-parameter regression (r up to 0.76) models involving MRI stiffness and pQCT BMD. Subjects who had at least one prevalent vertebral fracture showed decreased MRI stiffness (up to 17.9 %) and pQCT density (up to 34.2 %) at the distal extremities compared to the non-fracture group. DXA lumbar spine BMD T-score was not associated with vertebral deformities. CONCLUSIONS: The association between vertebral deformities and distal extremity measures supports the notion of postmenopausal osteoporosis as a systemic phenomenon.


Subject(s)
Osteoporosis, Postmenopausal/complications , Osteoporotic Fractures/etiology , Radius/pathology , Spinal Curvatures/etiology , Spinal Fractures/etiology , Tibia/pathology , Aged , Aged, 80 and over , Bone Density/physiology , Female , Humans , Magnetic Resonance Imaging/methods , Middle Aged , Osteoporosis, Postmenopausal/diagnosis , Osteoporosis, Postmenopausal/physiopathology , Osteoporotic Fractures/physiopathology , Radius/diagnostic imaging , Radius/physiopathology , Retrospective Studies , Spinal Curvatures/physiopathology , Spinal Fractures/physiopathology , Tibia/diagnostic imaging , Tibia/physiopathology , Tomography, X-Ray Computed/methods
7.
Clin Exp Immunol ; 171(3): 319-29, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23379439

ABSTRACT

Diabetes is associated with an increased risk of death from infectious disease. Hyperglycaemia has been identified as the main factor contributing to the development of diseases associated with diabetes mellitus. However, experimental evidence indicates individual susceptibility to develop complications of diabetes. In this context, the aim of this work was to study the immune response in a streptozotocin-induced type 1 diabetes in two mouse strains: BALB/cByJ and C57Bl/6J. The participation of hyperglycaemia and oxidative stress was also analysed. Diabetic BALB/cByJ mice showed a decrease in both the in-vivo and in-vitro immune responses, whereas diabetic C57Bl/6J mice had higher blood glucose but exhibited no impairment of the immune response. The influence of hyperglycaemia over the immune response was evaluated by preincubation of lymphocytes from normal mice in a high glucose-containing medium. T and B cells from BALB/cByJ mice showed a decrease in cell viability and mitogen-stimulated proliferation and an increase in apoptosis induction. An increase in oxidative stress was implicated in this deleterious effect. These parameters were not affected in the T and B lymphocytes from C57Bl/6J mice. In conclusion, BALB/cByJ mice were sensitive to the deleterious effect of hyperglycaemia, while C57BL/6J were resistant. Although an extrapolation of these results to clinical conditions must be handled with caution, these results highlight the need to contemplate the genetic background to establish models to study the deleterious effect of diabetes in order to understand phenotypical variations that are of clinical importance in the treatment of patients.


Subject(s)
Diabetes Mellitus, Experimental/immunology , Hyperglycemia/immunology , Oxidative Stress , Animals , Blood Glucose/analysis , Diabetes Mellitus, Experimental/metabolism , Female , Glutathione/metabolism , Lymphocyte Activation , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Streptozocin
8.
Stress ; 13(5): 384-91, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20666647

ABSTRACT

Stress, an important aspect of modern life, has long been associated with an altered homeostatic state. Little is known about the effect of the life stress on the outcome of diabetes mellitus, especially related to the higher risk of infections. Here, we evaluate the effects of chronic mild stress (CMS) exposure on the evolution of type I diabetes induced by streptozotocin administration in BALB/c mice. Exposure of diabetic mice to CMS resulted in a significant reduction of survival and a sustained increase in blood glucose values. Concerning the immune response, chronic stress had a differential effect in mice with diabetes with respect to controls, showing a marked decrease in both T- and B-cell proliferation. No correlation was found between splenic catecholamine or circulating corticosterone levels and the proliferative response. However, a significant negative correlation was found between glucose levels and concanavalin A- and lipopolysaccharide-stimulated proliferative responses of T and B cells. A positive correlation between blood glucose and splenic catecholamine concentrations was found in diabetic mice but not in controls subjected to CMS. Hence, the present report shows that diabetic mice show a worse performance in immune function after stress exposure, pointing to the importance of considering life stress as a risk factor for patients with diabetes.


Subject(s)
Diabetes Mellitus, Experimental/blood , Diabetes Mellitus, Experimental/psychology , Hormones/physiology , Hyperglycemia/blood , Stress, Psychological/immunology , Animals , B-Lymphocytes/drug effects , B-Lymphocytes/immunology , Blood Glucose/metabolism , Catecholamines/blood , Cells, Cultured , Chronic Disease , Corticosterone/blood , Female , Food Deprivation , Mice , Mice, Inbred BALB C , Mitogens/pharmacology , Survival Analysis , T-Lymphocytes/drug effects , T-Lymphocytes/immunology , Water Deprivation/physiology
9.
J Neuroimmunol ; 218(1-2): 12-20, 2010 Jan 25.
Article in English | MEDLINE | ID: mdl-19942299

ABSTRACT

This work shows a comparative study on the effects of chronic mild stress upon learning and memory and immunity, in BALB/c and C57BL/6 mice. Stressed BALB/c, but not C57Bl/6 mice, showed a poor learning performance, morphological alterations in the hippocampus with an increase in oxidative stress. A correlation between poor memory performance and the increase of the Th2/Th1 balance was found. Our results suggest that vulnerability to cognitive deficit associated with stress exposition could be related to a differential regulation of Th1/Th2 cytokine balance, suggesting a better learning performance for individuals that produce Th1 type cytokine after stress exposition.


Subject(s)
Cognition Disorders/immunology , Cognition Disorders/psychology , Genetic Predisposition to Disease , Stress, Psychological/immunology , Th1 Cells/immunology , Th2 Cells/immunology , Animals , Cell Separation , Cognition Disorders/genetics , Cytokines/immunology , Flow Cytometry , Hippocampus/immunology , Hippocampus/pathology , Maze Learning , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Oxidative Stress/immunology , Reactive Oxygen Species/immunology , Reactive Oxygen Species/metabolism , Stress, Psychological/genetics
10.
Magn Reson Med ; 61(5): 1114-21, 2009 May.
Article in English | MEDLINE | ID: mdl-19215044

ABSTRACT

Fast large-angle spin echo (FLASE) is a common pulse sequence designed for quantitative imaging of trabecular bone (TB) microarchitecture. However, imperfections in the nonselective phase-reversal pulse render it prone to stimulated echo artifacts. The problem is further exacerbated at isotropic resolution. Here, a substantially improved RF-spoiled FLASE sequence (sp-FLASE) is described and its performance is illustrated with data at 1.5T and 3T. Additional enhancements include navigator echoes for translational motion sensing applied in a slice parallel to the imaging slab. Whereas recent work suggests the use of fully-balanced FLASE (b-FLASE) to be advantageous from a signal-to-noise ratio (SNR) point of view, evidence is provided here that the greater robustness of sp-FLASE may outweigh the benefits of the minor SNR gain of b-FLASE for the target application of TB imaging in the distal extremities, sites of exclusively fatty marrow. Results are supported by a theoretical Bloch equation analysis and the pulse sequence dependence of the effective T(2) of triglyceride protons. Last, sp-FLASE images are shown to provide detailed and reproducible visual depiction of trabecular networks in three dimensions at both anisotropic (137 x 137 x 410 microm(3)) and isotropic (160 x 160 x 160 microm(3)) resolutions in the human distal tibia in vivo.


Subject(s)
Algorithms , Chronic Kidney Disease-Mineral and Bone Disorder/pathology , Echo-Planar Imaging/methods , Image Enhancement/methods , Image Interpretation, Computer-Assisted/methods , Imaging, Three-Dimensional/methods , Tibia/pathology , Humans , Reproducibility of Results , Sensitivity and Specificity , Spin Labels
11.
Clin Exp Immunol ; 154(2): 235-46, 2008 Nov.
Article in English | MEDLINE | ID: mdl-18778365

ABSTRACT

Diabetes is widely believed to predispose to serious infections. However, the mechanisms linking diabetes and immunosuppression are not well defined. One potential mediator of the altered defence mechanisms is hyperglycaemia. It has been identified as the main factor contributing to the development of diseases associated with diabetes mellitus. In this study we analyse the immune response in diabetes and the direct effect of hyperglycaemia on T and B lymphocyte reactivity. Diabetes induced an early decrease in IgG levels in the secondary response. However, both primary responses against a T-cell-dependent or independent antigen were affected after 6 months of diabetes induction. T- and B- cell proliferation was only decreased at this time. To gain insight into the potential mechanisms involved, we evaluated the influence of hyperglycaemia over the immune response. Pre-incubation of lymph node and spleen cells in a high glucose (HG) containing medium led to a significant time- and dose-dependent decrease in T- and B-cell proliferation. This effect was associated with the presence of HG-derived supernatants. Still viable cells after HG exposition were able to improve their proliferative response when cultured with the mitogen in a fresh standard medium. HG diminished cell viability, increased apoptosis and induced oxidative stress in lymphocytes. These results indicate that HG concentrations can directly affect lymphoid cell growth. An increase in oxidative stress would be implicated in this deleterious effect. The possibility that prolonged exposure to pathologically HG concentrations would result in the immunosuppressive state observed in diabetes is also discussed.


Subject(s)
Diabetes Mellitus, Experimental/immunology , Diabetes Mellitus, Type 1/immunology , Hyperglycemia/immunology , Animals , Apoptosis/drug effects , B-Lymphocytes/drug effects , B-Lymphocytes/immunology , Cell Proliferation/drug effects , Cells, Cultured , Dose-Response Relationship, Immunologic , Female , Glucose/pharmacology , Immunoglobulin G/biosynthesis , Immunoglobulin M/biosynthesis , Lipid Peroxidation/drug effects , Lymphocyte Activation/drug effects , Lymphocyte Activation/immunology , Mice , Mice, Inbred BALB C , Mitogens/immunology , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolism , T-Lymphocytes/drug effects , T-Lymphocytes/immunology
12.
Clin Exp Immunol ; 153(2): 289-96, 2008 Aug.
Article in English | MEDLINE | ID: mdl-18549441

ABSTRACT

The aim of the present study was to assess the effect of dehydroepiandrosterone (DHEA: 10 microM) and metformin (10 microM and 100 microM) in regulating proliferation of cultured T lymphocytes. T cells were isolated from lymph nodes of prepuberal BALB/c mice. We found that DHEA, metformin and DHEA + metformin added to the incubation media diminished proliferation of T cells. The inhibition by DHEA was higher than that produced by metformin, while the combined treatment showed a synergistic action that allowed us to speculate distinct regulatory pathways. This was supported later by other findings in which the addition of DHEA to the incubation media did not modify T lymphocyte viability, while treatment with metformin and DHEA + metformin diminished cellular viability and increased both early and late apoptosis. Moreover, DHEA diminished the content of the anti-oxidant molecule glutathione (GSH), whereas M and DHEA + metformin increased GSH levels and diminished lipid peroxidation. We conclude that DHEA and metformin diminish proliferation of T cells through different pathways and that not only the increase, but also the decrease of oxidative stress inhibited proliferation of T cells, i.e. a minimal status of oxidative stress, is necessary to trigger cellular response.


Subject(s)
Antioxidants/pharmacology , Dehydroepiandrosterone/pharmacology , Metformin/pharmacology , T-Lymphocytes/cytology , Animals , Apoptosis/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Female , Glutathione/metabolism , Lipid Peroxidation/drug effects , Lymphocyte Activation , Mice , Mice, Inbred BALB C , Necrosis , Nitric Oxide Synthase/metabolism , Oxidative Stress/drug effects , Polycystic Ovary Syndrome/immunology , T-Lymphocytes/drug effects , T-Lymphocytes/enzymology
13.
J Safety Res ; 39(3): 269-72, 2008.
Article in English | MEDLINE | ID: mdl-18571567

ABSTRACT

PROBLEM: Among older adults, both unintentional falls and traumatic brain injuries (TBI) result in significant morbidity and mortality; however, only limited national data on fall-related TBI are available. METHOD: To examine the relationship between older adult falls and TBI deaths and hospitalizations, CDC analyzed 2005 data from the National Center for Health Statistics' National Vital Statistics System and the Agency for Healthcare Research and Quality's Nationwide Inpatient Sample. RESULTS: In 2005, among adults>or=65 years, there were 7946 fall-related TBI deaths and an estimated 56,423 hospitalizations for nonfatal fall-related TBI in the United States. Fall-related TBI accounted for 50.3% of unintentional fall deaths and 8.0% of nonfatal fall-related hospitalizations. SUMMARY: These findings underscore the need for greater dissemination and implementation of evidence-based fall prevention interventions.


Subject(s)
Accidental Falls/mortality , Brain Injuries/mortality , Aged , Aged, 80 and over , Female , Hospitalization/statistics & numerical data , Humans , Male , United States/epidemiology
14.
Int J Impot Res ; 20(2): 145-9, 2008.
Article in English | MEDLINE | ID: mdl-18075506

ABSTRACT

Erectile dysfunction is a prevalent disorder that not only affects men with the disorder but also their partners. Significant improvements in the sexual health of these couples have been achieved with the introduction of phosphodiesterase 5 (PDE5) inhibitors. Currently PDE5 inhibitors are used on an on-demand basis. New evidence regarding the effects of PDE5 inhibitors on the underlying pathophysiologic processes that cause erectile dysfunction have sparked interest in the continuous dosing of these medications. We will discuss the biological background and the available clinical evidence for the continuous use of phosphodiesterase inhibitors in erectile dysfunction. Lastly, we will discuss the emerging clinical data for the use of daily PDE5 inhibitors in men with lower urinary tract symptoms.


Subject(s)
Erectile Dysfunction/drug therapy , Erectile Dysfunction/physiopathology , Phosphodiesterase Inhibitors/administration & dosage , Urination Disorders/drug therapy , Drug Administration Schedule , Humans , Male , Treatment Outcome
15.
Int Arch Allergy Immunol ; 145(3): 193-206, 2008.
Article in English | MEDLINE | ID: mdl-17912007

ABSTRACT

BACKGROUND: Recombinant allergens and especially their hypoallergenic variants are promising candidates for a more effective and safer specific immunotherapy. METHODS: Physicochemical and immunological characteristics of a folding variant of recombinant Bet v 1 (rBet v 1-FV) were investigated in comparison to natural Bet v 1 (nBet v 1) and the correctly folded recombinant Bet v 1 (rBet v 1-WT) by SDS-PAGE, size exclusion chromatography, multi-angle light scattering, circular dichroism, immunoblotting and enzyme allergosorbent test inhibition assay for detection of IgE reactivity and ELISA with Bet v 1-specific monoclonal antibodies. The functional IgE reactivity of the different Bet v 1 proteins was investigated using basophil activation in terms of CD203c expression and histamine release. T cell reactivity was investigated using T cell lines raised from birch pollen-allergic subjects against nBet v 1. Immunogenicity was investigated in mice. RESULTS: Physicochemical characterization revealed purity, homogeneity and monomeric properties of rBet v 1-FV. Unlike nBet v 1 and rBet v 1-WT, rBet v 1-FV showed almost no IgE binding in immunoblots. The reduction of allergenicity was further proved by IgE-binding inhibition assays, basophil activation and histamine release. T cell reactivity was completely conserved, as demonstrated by proliferation of Bet v 1-specific T cell lines with multiple epitope specificities. rBet v 1-FV showed strong immunogenicity in mice. CONCLUSIONS: Due to its reduced IgE reactivity and decreased capacity to activate basophils, but retained T cell reactivity and strong immunogenicity, rBet v 1-FV proved to be a very promising candidate for specific immunotherapy in birch pollen-allergic subjects.


Subject(s)
Allergens/immunology , Betula/immunology , Desensitization, Immunologic/methods , Pollen/immunology , Recombinant Proteins/immunology , Adolescent , Adult , Aged , Allergens/metabolism , Allergens/therapeutic use , Animals , Antibody Specificity , Basophils/immunology , Drug Evaluation, Preclinical , Epitopes/immunology , Female , Humans , Immune Sera/immunology , Immunoglobulin E/immunology , Lymphocyte Activation , Male , Mice , Mice, Inbred BALB C , Middle Aged , Protein Folding , Recombinant Proteins/metabolism , Recombinant Proteins/therapeutic use , Rhinitis, Allergic, Seasonal/immunology , Rhinitis, Allergic, Seasonal/therapy , T-Lymphocytes/immunology
16.
Clin Exp Allergy ; 37(3): 441-50, 2007 Mar.
Article in English | MEDLINE | ID: mdl-17359394

ABSTRACT

BACKGROUND: Immunotherapy of grass pollen allergy is currently based on the administration of pollen extracts containing natural allergens. Specifically designed recombinant allergens with reduced IgE reactivity could be used in safer and more efficacious future therapy concepts. OBJECTIVES: This study aimed to generate hypoallergenic variants of the timothy grass major allergen Phl p 5a as candidates for allergen-specific immunotherapy. METHODS: Three deletion mutants were produced in Escherichia coli and subsequently purified. The overall IgE-binding capacity of the mutants was compared with the recombinant wild-type allergen by membrane blot and IgE-inhibition assays. The capacity for effector cell activation was determined in basophil activation assays. T cell proliferation assays with allergen-specific T cell lines were performed to confirm the retention of T cell reactivity. Structural properties were characterized by circular dichroism analysis and homogeneity by native isoelectric focusing. The deletion sites were mapped on homology models comprising the N- and C-terminal halves of Phl p 5a, respectively. RESULTS: The double-deletion mutant rPhl p 5a Delta(94-113, 175-198) showed strongly diminished IgE binding in membrane blot and IgE-inhibition assays. Both deletions affect predominantly alpha-helical regions located in the N- and C-terminal halves of Phl p 5a, respectively. Whereas deletion of Delta175-198 alone was sufficient to cause a large reduction of the IgE reactivity in a subgroup of allergic sera, only the combination of both deletions was highly effective for all the sera tested. rPhl p 5a Delta(94-113, 175-198) consistently showed at least an 11.5-fold reduced capacity to activate basophils compared with the recombinant wild-type molecule, and the T cell proliferation assays demonstrated retention of T cell reactivity. CONCLUSION: The mutant rPhl p 5a Delta(94-113, 175-198) fulfils the basic requirements for a hypoallergenic molecule suitable for a future immunotherapy of grass pollen allergy; it offers substantially reduced IgE binding and maintained T cell reactivity.


Subject(s)
Allergens/chemistry , Immunoglobulin E/therapeutic use , Plant Proteins/chemistry , Rhinitis, Allergic, Seasonal/drug therapy , Allergens/immunology , Allergens/therapeutic use , Female , Humans , Male , Plant Proteins/immunology , Plant Proteins/therapeutic use , Pollen/immunology
17.
Indian J Pathol Microbiol ; 50(4): 764-5, 2007 Oct.
Article in English | MEDLINE | ID: mdl-18306545

ABSTRACT

A rare case of primary mucinous eccrine carcinoma of the lower eyelid in a 45 year old female is described. The lesion was diagnosed clinically as melanoma. We report this case because of unusual nature of this tumor and rare presentation in a female patient.


Subject(s)
Adenocarcinoma, Mucinous/diagnosis , Adenocarcinoma, Mucinous/pathology , Eccrine Glands/pathology , Eyelid Neoplasms/diagnosis , Eyelid Neoplasms/pathology , Eyelids/pathology , Female , Humans , Melanoma/diagnosis , Middle Aged
18.
Intensive Care Med ; 31(8): 1095-100, 2005 Aug.
Article in English | MEDLINE | ID: mdl-15999252

ABSTRACT

OBJECTIVE: Volutrauma caused by high tidal volumes contributes considerably to the development of bronchopulmonary dysplasia. Yet high tidal volumes are required to overcome dead space. In an experimental arrangement we tested whether reduction of dead space might reduce ventilation requirements and thus reduce volutrauma in preterm infants. MATERIALS AND METHODS: The time required to eliminate CO2 by standardized mechanical ventilation from a preterm infant's test lung flooded with CO2 was measured. Four different Y-pieces and flow sensor combinations were tested with and without a device for closed suction: Y-piece without flow sensor; integrated flow sensor; small dead-space flow sensor; and a new dead-space free-flow sensor for preterm infants. CO2 concentrations were measured by a capnograph. Mean CO2 elimination times (+/-SD) were compared. RESULTS: Mean CO2 elimination time was 37.5 s (+/-1.18 s) with and 37.4 s (+/-0.97 s) without closed suction device for the Y-piece without flow sensor, 47.7 s (+/-0.82 s) and 45.5 s (+/-1.18 s) for the integrated flow sensor, 42.5 s (+/-1.27 s) and 41.1 s (+/-0.99 s) for the small dead-space flow sensor and 38.3 s (+/-1.16 s) and 36.8 s (+/-0.79 s) for the dead-space free-flow sensor. CONCLUSION: CO2 elimination time with and without closed suction device was nearly identical for the Y-piece without flow sensor and for the dead-space free-flow sensor. With both systems, ventilation requirements were significantly lower than for the integrated flow sensor and for the small dead-space flow sensor (integrated flow sensor vs dead-space free-flow sensor 23.6 and 24.5%, respectively, small dead-space flow sensor vs dead-space free flow sensor 11.7 and 10.9%, respectively); thus, we think that introduction of the innovative dead-space free-flow sensor into clinical practice might reduce incidence and severity of bronchopulmonary dysplasia by reduction of volutrauma.


Subject(s)
Pulmonary Ventilation , Respiration, Artificial/instrumentation , Bronchopulmonary Dysplasia/therapy , Carbon Dioxide/analysis , Humans , Infant, Newborn , Infant, Very Low Birth Weight , Intensive Care Units, Neonatal , Maximal Voluntary Ventilation , Respiratory Dead Space
19.
Biol Trace Elem Res ; 104(2): 173-83, 2005 May.
Article in English | MEDLINE | ID: mdl-15894817

ABSTRACT

Zinc and iron are crucial mineral components of human diet, because their deficiency leads to several disorders, including alterations of the immune function. It has been demonstrated, in both humans and rodents, that a diminished number of lymphoid cells and a loss of lymphocyte activity accompany deprivation of these essential minerals. The aim of this work was to analyze if iron and/or zinc imbalances regulate lymphocyte activity and the intracellular signals involved in the effect. Mice from the BALB/c strain were fed with iron- and/or zinc-deficient or mineral-supplemented diets, according to the American Institute of Nutrition Rodent Diets. Levels of iron and zinc were assessed in blood, liver, or bone samples. Selective mitogen stimulation of T- and B-lymphocytes were performed. We found a diminished proliferative response in T- and B-lymphocytes from zinc- and/or iron-deficient animals with respect to controls. These effects were related to decreased mitogen-induced translocation of protein kinase C (PKC) activity to cell membranes on both cell types from all animals fed with deficient diets. Our results demonstrate that iron and zinc deficiencies affect both T- and B-lymphocyte function by PKC-dependent mechanisms.


Subject(s)
B-Lymphocytes/immunology , Iron Deficiencies , Protein Kinase C/physiology , T-Lymphocytes/immunology , Zinc/deficiency , Animals , Cell Proliferation/drug effects , Concanavalin A/pharmacology , Female , Lipopolysaccharides/pharmacology , Mice , Mice, Inbred BALB C , Phytohemagglutinins/pharmacology , Pokeweed Mitogens/pharmacology
20.
Acta Paediatr ; 93(9): 1166-71, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15384878

ABSTRACT

AIM: A central respiratory regulation disturbance--triggered by impaired oxygen supply to the brainstem--is being discussed as an aetiological factor in sudden infant death syndrome. In this experimental study, further insight was sought into how far cervical spine movements may induce critical narrowing of the vertebral arteries. METHODS: In 10 infant bodies, the vertebral arteries were cannulated close to their origin and perfused with a fluid bolus of 5 ml in 2 s. The intravascular peak pressure created was memorized. Individual resistance indices were computed for every vertebral artery by relating the average pressure maximum from 10 measurements each with maximal rotation of the neck to either side alone and then with additional hyperextension of the cervical spine to the average peak pressure in neutral head position. RESULTS: Alterations of position-induced pressure changes occurred in the vertebral arteries of all children. Considering exclusively the combined movements of rotation plus extension, resistance increased ipsi- and contralaterally--no matter which side the head was turned--in three infants. A further three reacted with resistance surges only contralateral to the direction of rotation, and one only ipsilateral. Double contralateral associated with only one-directional ipsilateral rise was observed twice, and vice versa once. CONCLUSION: The presented method enables crude quantification of postmortem dynamic resistance alterations in the vertebral arteries. How far such measurements reflect authentic circulatory conditions during life remains to be assessed.


Subject(s)
Autopsy/methods , Blood Pressure/physiology , Posture/physiology , Vascular Resistance/physiology , Vertebral Artery/physiopathology , Female , Head Movements/physiology , Humans , Infant , Infant, Newborn , Male , Sudden Infant Death/etiology
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