ABSTRACT
Indirect searches for dark matter through Standard Model products of its annihilation generally assume a cross-section which is dominated by a term independent of velocity (s-wave annihilation). However, in many DM models an s-wave annihilation cross-section is absent or helicity suppressed. To reproduce the correct DM relic density in these models, the leading term in the cross section is proportional to the DM velocity squared (p-wave annihilation). Indirect detection of such p-wave DM is difficult because the average velocities of DM in galaxies today are orders of magnitude slower than the DM velocity at the time of decoupling from the primordial thermal plasma, thus suppressing the annihilation cross-section today by some five orders of magnitude relative to its value at freeze out. Thus p-wave DM is out of reach of traditional searches for DM annihilations in the Galactic halo. Near the region of influence of a central supermassive black hole, such as Sgr A*, however, DM can form a localized over-density known as a "spike". In such spikes the DM is predicted to be both concentrated in space and accelerated to higher velocities, thereby allowing the γ-ray signature from its annihilation to potentially be detectable above the background. We use the Fermi Large Area Telescope to search for the γ-ray signature of p-wave annihilating DM from a spike around Sgr A* in the energy range 10 GeV-600 GeV. Such a signal would appear as a point source and would have a sharp line or box-like spectral features difficult to mimic with standard astrophysical processes, indicating a DM origin. We find no significant excess of γ rays in this range, and we place upper limits on the flux in γ-ray boxes originating from the Galactic Center. This result, the first of its kind, is interpreted in the context of different models of the DM density near Sgr A*.
ABSTRACT
Giant omphalocele is associated to morbidity and mortality because of the strain the reintegrated herniated mass places on the hemodynamic equilibrium and breathing functions of affected infants. Currently, care management consists in a reintegration in one time or progressive reintegration. We report here a multicenter retrospective study about alternative management by VAC® therapy for giant omphaloceles. The study included three patients (1 girl, 2 boys) presenting with giant omphaloceles, born at full term in three different University Hospitals (prenatal diagnosis, normal karyotype). VAC® therapy was implemented at different times according to the cases (at Day 11, Month 1 and Month 5 after birth). The initial pressure applied was -10 mmHg progressively increased to -50 mmHg. A middle size VAC GranuFoam Silver® Dressing was used in all cases. Wound healing occurred at Month 4 for the first case, Month 6 and Month 8 for the other two. VAC® therapy is a good alternative for the care management of giant omphaloceles with more advantages especially when using prosthetic material. We also aimed at refining the most adapted indications in these specific situations, and finally we envisioned a harmonization of care for these children.
Subject(s)
Negative-Pressure Wound Therapy , Female , Hernia, Umbilical , Humans , Infant, Newborn , Male , Negative-Pressure Wound Therapy/methods , Retrospective Studies , Wound HealingABSTRACT
BACKGROUND: Currently there is no drug proven to effectively treat cats with feline infectious peritonitis (FIP). HYPOTHESIS: Propentofylline (PPF) can decrease vasculitis, and therefore prolong survival time in cats with FIP, and increase their quality of life. ANIMALS: Twenty-three privately owned cats with FIP. METHODS: Placebo-controlled double-blind trial. FIP was confirmed by histology or immunostaining of feline coronavirus (FCoV) antigen in effusion or tissue macrophages or both. The cats were randomly selected for treatment with either PPF or placebo. All cats received additional treatment with glucocorticoids, antibiotics, and low molecular weight heparin according to methods. RESULTS: There was no statistically significant difference in the survival time of cats treated with PPF (8 days, 95% CI 5.4-10.6) versus placebo (7.5 days, 95% CI 4.4-9.6). The median survival time of all cats was 8 days (4-36 days). There was neither a difference in quality of life (day 7, P = .892), in the amount of effusion (day 7, P = .710), the tumor necrosis factor-alpha (TNF-α) concentration (day 7, P = .355), nor in any other variable investigated in this study, including a complete blood count, and a small animal biochemistry profile. CONCLUSIONS AND CLINICAL IMPORTANCE: This study did not detect an effect of PPF on the survival time, the quality of life, or any clinical or laboratory parameter in cats with FIP. Therefore, PPF does not appear to be an effective treatment option in cats with a late stage of the disease FIP.
Subject(s)
Anti-Ulcer Agents/therapeutic use , Feline Infectious Peritonitis/drug therapy , Xanthines/therapeutic use , Animals , Cats , Feline Infectious Peritonitis/blood , Quality of Life , Tumor Necrosis Factor-alpha/bloodABSTRACT
Aggressive angiomyxoma (AAM) is a rare tumor that preferentially involves the pelvis and perineal regions and arises from the connective tissue. Its cause and pathogenesis are unknown at present. Treatment typically involves surgery, and despite apparently complete resection, local recurrences are common. We describe a case of a large angiomyxoma of the left pelvis in a 59-year-old woman who underwent two surgical excisions. The first had been done in May 1998. She developed a local recurrence in December 1998. A palliative resection with macroscopic residuals was performed in February 2001, followed by radiation therapy with a total dose of 60 Gy. The diagnosis was revised at the time of the second operation. Initially, the tumor was diagnosed as angiomyofibroblastoma. Follow-up 3 years after the radiation treatment revealed no recurrence. The time of the local control achieved as yet is already longer than the former time to progression between the first two surgical procedures. This is, to our knowledge, the second description of a therapeutic irradiation of a recurrent AAM. Radiation therapy is able to control a recurrent AAM for at least 3 years.
Subject(s)
Myxoma/radiotherapy , Neoplasm Recurrence, Local/radiotherapy , Pelvic Neoplasms/radiotherapy , Retroperitoneal Neoplasms/radiotherapy , Buttocks , Female , Humans , Middle Aged , Myxoma/pathology , Myxoma/surgery , Neoplasm Invasiveness , Neoplasm Recurrence, Local/pathology , Pelvic Neoplasms/pathology , Pelvic Neoplasms/surgery , Perineum , Reoperation , Retroperitoneal Neoplasms/pathology , Retroperitoneal Neoplasms/surgeryABSTRACT
Originally defined by their patterns of cytokine production, Th1 and Th2 cells have been described more recently to express other genes differentially as well, at least in vitro. In this study we compared the expression of Th1- and Th2-associated genes directly during in vivo sensitization to ovalbumin (OVA) in Th1- and Th2-polarized models of airways inflammation. Th1-polarized airway inflammation was achieved by the intranasal instillation of adenoviral vectors (Ad) encoding granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-12, followed by daily aerosolizations of OVA; instillation of Ad/GM-CSF alone with OVA aerosolization led to Th2-polarized responses. Lymph nodes were obtained at various time-points, RNA extracted, and analysed by real-time quantitative polymerase chain reaction (PCR). Consistent with reports from in vitro and human studies, mice undergoing Th1-polarized inflammation showed preferential expression of the transcription factor t-bet, the chemokines IFN-gamma inducible protein (IP)-10 and macrophage inflammatory protein 1 alpha (MIP-1-alpha), and the chemokine receptor CCR5. In contrast, the transcription factor GATA-3, the chemokines I-309 and thymus and activation regulated chemokine (TARC), and the chemokine receptors CCR3 and CCR4 were preferentially expressed in the Th2 model. Importantly, we also show that Ad/transgene expression remains compartmentalized to the lung after intranasal instillation. Flow cytometric analysis of lung myeloid dendritic cells indicated that B7.1 was expressed more strongly in the Th1 model than in the Th2 model. These studies provide a direct comparison of gene expression in in vivo Th1- and Th2-polarized models, and demonstrate that molecular events in the lymph nodes can be altered fundamentally by cytokine expression at distant mucosal sites.
Subject(s)
Cytokines/analysis , Lung/immunology , Lymph Nodes/immunology , Th1 Cells/immunology , Th2 Cells/immunology , Animals , Chemokines/analysis , Chemokines/immunology , Cytokines/immunology , DNA-Binding Proteins/analysis , Dendritic Cells/immunology , Female , Flow Cytometry/methods , GATA3 Transcription Factor , Gene Expression/immunology , Mice , Mice, Inbred BALB C , Ovalbumin/immunology , Polymerase Chain Reaction/methods , Receptors, Chemokine/analysis , Receptors, Chemokine/immunology , Respiratory Hypersensitivity/immunology , Respiratory Mucosa/immunology , T-Box Domain Proteins , Thorax/immunology , Trans-Activators/analysis , Transcription Factors/analysis , Transgenes/genetics , Transgenes/immunologyABSTRACT
BACKGROUND: Exposure to aerosolized harmless antigen such as ovalbumin (OVA) has previously been shown to induce inhalation tolerance, a state characterized by inhibition of IgE synthesis and airway inflammation, upon secondary immunogenic antigen encounter. Immune events associated with this phenomenon are still poorly understood. OBJECTIVE: The aim of this study was to investigate cellular and molecular mechanisms underlying this state of 'unresponsiveness'. METHODS: After initial repeated OVA exposure, mice were subjected to a protocol of antigen-induced airway inflammation, encompassing two intraperitoneal injections of OVA adsorbed to aluminium hydroxide followed by airway challenge. We assessed immune events in the draining lymph nodes after sensitization, and in the lungs after challenge. RESULTS: In animals initially exposed to OVA, we observed, at the time of sensitization, considerable expansion of T cells, many of which expressed the activation markers CD69 and CD25, as well as increased numbers of antigen-presenting cells, particularly B cells. While these animals produced low levels of IgE, the observed elevated levels of IgG1 signified isotype switching. Splenocytes and lymph node cells from OVA-exposed mice produced low levels of IL-4, IL-5, IL-13 and IFN-gamma, indicating aborted effector function of both T helper (Th)2- and Th1-associated cytokines. Real time quantitative polymerase chain reaction (PCR) (TaqMan) analysis of costimulatory molecules in the lungs after in vivo challenge showed that B7.1, B7.2, CD28 and CTLA-4 mRNA expression was low in animals initially exposed to OVA. Ultimately, these events were associated with abrogated airway inflammation and attenuated airway hyper-responsiveness. The decreased inflammation was antigen-specific and independent of IL-10 or IFN-gamma. CONCLUSION: Initial exposure to OVA establishes a programme that prevents the generation of intact, fully functional inflammatory responses upon secondary antigen encounter. The absence of inflammation, however, is not associated with categorical immune unresponsiveness.
Subject(s)
Cytokines/drug effects , Cytokines/immunology , Immunoglobulins/drug effects , Immunoglobulins/immunology , Inhalation Exposure/adverse effects , Mice, Inbred BALB C/immunology , Ovalbumin/immunology , Ovalbumin/pharmacology , Animals , Antibody Specificity/drug effects , Antibody Specificity/immunology , Biomarkers/blood , Bronchial Provocation Tests , Bronchoalveolar Lavage Fluid/chemistry , Bronchoalveolar Lavage Fluid/immunology , CD4-Positive T-Lymphocytes/immunology , CD4-Positive T-Lymphocytes/metabolism , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/metabolism , Cytokines/blood , Dose-Response Relationship, Immunologic , Female , Histocompatibility Antigens Class II/drug effects , Histocompatibility Antigens Class II/immunology , Immunoglobulins/blood , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/metabolism , Lung/blood supply , Lung/cytology , Mice , Mice, Knockout , Models, Animal , RNA, Messenger/biosynthesis , RNA, Messenger/drug effects , Sodium Chloride/pharmacology , Time FactorsABSTRACT
The objective of this study was to define phenotypic changes of antigen-presenting cells (APCs) and T cells in a murine model of antigen-induced airways inflammation that involves intraperitoneal sensitization with ovalbumin (OVA)/adjuvant followed by antigen aerosolization. We investigated the APC and T-cell compartments both after sensitization (primary immune response) and after challenge (secondary immune response) at the thoracic lymph nodes (initiation site) and the lung (effector site). Our findings document a major cellular expansion in the lymph nodes after both sensitization and challenge. After sensitization, this expansion was comprised mainly of B cells, a considerable proportion of which expressed B7.2. At this time, T cells were markedly expanded and activated as assessed by CD69 expression; further, although GATA-3 and signal transducer and activator of transcription-6 were expressed at this time point, expression of interleukin (IL)-4, IL-5, and IL-13 messenger RNA (mRNA) levels were marginal. However, in vitro stimulation of lymph-node cells with OVA led to cytokine production. In contrast, 24 h after challenge, but not after sensitization, there was a major expansion of dendritic cells and macrophages in the lungs. This expansion was associated with enhanced expression of both B7.1 and B7.2. We also observed expansion of activated CD3(+)/CD4(+) T cells expressing the T helper-2-associated marker T1/ST2 in the lung, most notably 5 d after challenge. Further, IL-4, IL-5, and IL-13, but not interferon-gamma mRNA were expressed at high levels 3 h after challenge. This study helps to elucidate the "geography" of immune responses generated in a conventional murine model of allergic airways inflammation.
Subject(s)
Antigen-Presenting Cells/immunology , Membrane Proteins , Ovalbumin/immunology , Pneumonia/immunology , T-Lymphocyte Subsets/immunology , Aerosols , Animals , Antigen-Presenting Cells/cytology , Antigen-Presenting Cells/metabolism , Antigens, CD/metabolism , B7-1 Antigen/metabolism , B7-2 Antigen , Bronchial Provocation Tests , Cytokines/genetics , Cytokines/metabolism , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Disease Models, Animal , Female , Flow Cytometry , GATA3 Transcription Factor , Histocompatibility Antigens Class II/metabolism , Interleukin-1 Receptor-Like 1 Protein , Lung/cytology , Lymph Nodes/cytology , Membrane Glycoproteins/metabolism , Mice , Mice, Inbred BALB C , Ovalbumin/administration & dosage , Pneumonia/chemically induced , Proteins/genetics , Proteins/metabolism , Receptors, Interleukin , STAT6 Transcription Factor , T-Lymphocyte Subsets/metabolism , Trans-Activators/genetics , Trans-Activators/metabolismABSTRACT
Under normal circumstances the lung is in a state of immunologic homeostasis, a condition in which exposure to innocuous antigens does not lead to immune-inflammatory responses. This is the only reasonable solution to the dilemma faced by the lung: The need to interact with the external environment and the need to avoid responding to most of the environmental antigens to which it is exposed. In allergic diseases, such as asthma, this homeostasis is undermined, and immuneinflammatory responses to harmless aeroallergens are activated. We describe the changes in antigen presentation and cellular activation observed in a model of allergic airway inflammation. Further, we present a summary of our work that investigated the impact of the airway cytokine microenvironment on the development of immune responses in the respiratory tract.
Subject(s)
Asthma/immunology , Animals , Antibody Formation , Antigen Presentation , Disease Models, Animal , Lymphocyte Activation , Mice , T-Lymphocytes/immunologyABSTRACT
Infantile patients with acid maltase deficiency have severe hypertrophic cardiomyopathy, left ventricular outflow obstruction, and generalized muscle weakness and die before 1 year of age. We identified 12 infants with acid maltase deficiency who had a similar clinical presentation but less severe cardiomyopathy and absence of left ventricular outflow obstruction, and 9 of 12 had longer survival with assisted ventilation and supplemental intubation.
Subject(s)
Glycogen Storage Disease Type II/classification , Glycogen Storage Disease Type II/diagnosis , Age of Onset , Female , Glycogen Storage Disease Type II/complications , Glycogen Storage Disease Type II/mortality , Glycogen Storage Disease Type II/pathology , Humans , Infant , Male , New York City/epidemiology , PrognosisABSTRACT
The objective of this study was to investigate the effect of airway gene transfer of interleukin (IL)-10, a cytokine with potent anti-inflammatory and immunoregulatory activities, on allergic mucosal sensitization. We used a recently described murine model that involves repeated exposures to aerosolized ovalbumin (OVA), daily for 10 d, in the context of granulocyte macrophage colony-stimulating factor (GM-CSF) expression in the airway environment achieved by intranasal delivery of a replication-deficient adenovirus carrying the GM-CSF transgene. The resulting inflammatory response was characterized by a T-helper 2 cytokine profile and marked airway eosinophilia. After complete resolution of the inflammatory response (Day 28), a single exposure to OVA reconstituted airway eosinophilia and induced airway hyperresponsiveness. We show that concurrent expression of IL-10 inhibited GM-CSF-driven OVA-specific inflammation in a dose-dependent manner. Specifically, IL-10 decreased the number of mononuclear cells, neutrophils, and eosinophils in the bronchoalveolar lavage fluid (BALF). Histologic evaluation of the tissue corroborated the findings in the BALF. Concurrent expression of IL-10 at the time of mucosal sensitization abrogated both the cellular and physiologic recall responses in vivo. Studies in interferon (IFN)-gamma knockout mice demonstrated that prevention of airway eosinophilia by IL-10 was IFN-gamma-independent and that expression of IL-10 was associated with decreased levels of IL-4, IL-5, and tumor necrosis factor-alpha in the BALF. Flow cytometric analysis of dispersed lung cells showed that expression of IL-10 in the airway reduced the absolute number of Class II major histocompatibility complex (MHC)(+)/CD11c(+) (dendritic cells) and Class II MHC(+)/Mac-1(bright) (macrophages) cells expressing the costimulatory molecules B7.1 and B7.2 by 30%. However, IL-10 coexpression did not prevent expansion of CD4 and CD8 T cells or expression of the early activation marker CD69 on T cells. Thus, airway gene transfer of IL-10 altered the immune response to OVA in a way that resulted in inhibition of airway inflammation. These findings suggest that development of an immunoregulatory strategy based on IL-10, alone or in combination with GM-CSF, warrants further consideration.
Subject(s)
Gene Transfer Techniques , Interleukin-10/genetics , Respiratory Hypersensitivity/drug therapy , Respiratory Mucosa/drug effects , Adenoviridae/genetics , Animals , Antigen-Presenting Cells/metabolism , Antigens, Surface/biosynthesis , Bronchoalveolar Lavage Fluid/cytology , Bronchoalveolar Lavage Fluid/immunology , Dose-Response Relationship, Drug , Eosinophilia/chemically induced , Eosinophilia/drug therapy , Eosinophilia/immunology , Female , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Granulocyte-Macrophage Colony-Stimulating Factor/immunology , Immunoglobulin E/blood , Interferon-gamma/genetics , Interleukin-10/biosynthesis , Mice , Mice, Inbred BALB C , Mice, Knockout , Ovalbumin/immunology , Respiratory Hypersensitivity/chemically induced , Respiratory Hypersensitivity/immunology , Respiratory Mucosa/immunology , T-Lymphocytes/metabolismABSTRACT
Expression of granulocyte macrophage colony-stimulating factor (GM-CSF) in the airway allows allergic sensitization to ovalbumin (OVA) in an experimental protocol that others have shown to induce inhalation tolerance. The ensuing response is characterized by T helper (Th)2 cytokines, marked eosinophilia in the bronchoalveolar lavage fluid (BALF) and the tissue, and goblet-cell hyperplasia. These findings, which underscore the importance of the airway microenvironment in the development of immune responses to airborne antigens, prompted us to investigate whether a Type 1 polarized cytokine milieu in the airway would modulate the allergic sensitization. To this end, we concurrently expressed GM-CSF and interleukin (IL)-12 in the airway, using an adenovirus-mediated gene transfer approach. Coexpression of IL-12 did not prevent the development of an antigen-specific immune inflammatory response, but altered its phenotype. Whereas a similar total cell number was observed in the BALF, airway eosinophilia was abrogated. Histologic evaluation of the tissue corroborated the findings in the BALF and demonstrated that IL-12 coexpression prevented goblet-cell hyperplasia. Expression of IL-12 decreased IL-4 and IL-5 content in the BALF by about 80 and 95%, respectively, and IL-5 in the serum by approximately 80%. In contrast, interferon (IFN)-gamma was increased in both BALF and serum. Similarly, we observed a Th2/Th1 shift in OVA-specific cytokine production in vitro. Recall challenge with OVA in vivo after resolution of the initial inflammatory response demonstrated that the effect of IL-12 was persistent. IL-12-mediated inhibition of airway eosinophilia was mainly IFN-gamma-independent, whereas inhibition of OVA-specific IgE synthesis was IFN-gamma-dependent. Our data underscore the importance of the airway microenvironment in the elicitation of immune responses to environmental antigens.
Subject(s)
Bronchi/immunology , Gene Transfer Techniques , Interleukin-12/genetics , Adenoviridae/genetics , Administration, Inhalation , Aerosols , Animals , Bronchi/metabolism , Bronchoalveolar Lavage Fluid/cytology , Cells, Cultured , Cytokines/immunology , Cytokines/pharmacology , Dose-Response Relationship, Drug , Female , Granulocyte-Macrophage Colony-Stimulating Factor/immunology , Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Immunoglobulin E/biosynthesis , Immunoglobulin E/immunology , Immunologic Memory/immunology , Interleukin-12/immunology , Interleukin-12/metabolism , Lung/metabolism , Mice , Mice, Inbred BALB C , Mice, Knockout , Mucous Membrane/immunology , Mucous Membrane/metabolism , Ovalbumin/administration & dosage , Ovalbumin/pharmacologyABSTRACT
BACKGROUND: Recent epidemiological studies have suggested that exposure to certain viruses and bacteria influences the development of allergy and allergic diseases, such as asthma. However, there is a paucity of experimental evidence examining the consequences of concurrent exposure to allergen and infectious agents, and the potential mechanisms by which allergic disease might be averted as a result. OBJECTIVE: To model this situation experimentally, we investigated whether a virally induced immune response, elicited by a replication-deficient human type 5 adenovirus (RDA) administered at a site distant from the airways, could inhibit ovalbumin (OVA)-induced airways eosinophilic inflammation. METHODS: C57BL/6 mice were infected intramuscularly with RDA 16h prior to intraperitoneal OVA sensitization. Cellular and cytokine responses in the lung/airways were examined after an OVA aerosol challenge. RESULTS: RDA infection significantly inhibited the inflammatory response in the lung tissue after antigen challenge. In the bronchoalveolar lavage (BAL), total cell number, eosinophils and lymphocytes were decreased by 70, 85 and 65%, respectively, after antigen challenge in RDA-treated, compared with untreated, mice. RDA infection had no effect on IgE synthesis. The levels of IL-5, IL-4 and IFNgamma in the BAL after antigen challenge were significantly lower in RDA-treated mice. In vitro production of cytokines by splenocytes in response to OVA restimulation revealed a shift from IL-4 in sensitized, PBS-treated mice, to IFNgamma in sensitized mice treated with RDA. Flow cytometric analysis revealed that RDA infection increased the proportion of CD8 T cells in the BAL; this change in T-cell subsets was accompanied by an increase in both CD4 and CD8 T cells positive for intracellular IFNgamma. Inhibition of antigen-induced airways inflammation was IFNgamma-dependent but did not require IL-12, as RDA-treatment inhibited airways inflammation in IL-12 but not IFNgamma knock-out mice. CONCLUSION: This study demonstrates that an immune response against a replication-deficient adenovirus during the initial exposure to OVA inhibits the development of airways inflammation after antigen aerosol challenge.
Subject(s)
Adenoviridae Infections/immunology , Adenoviruses, Human/immunology , Defective Viruses/immunology , Eosinophils/immunology , Lung/immunology , Respiratory Hypersensitivity/immunology , Animals , Bronchoalveolar Lavage Fluid/cytology , Bronchoalveolar Lavage Fluid/immunology , Cytokines/biosynthesis , Female , Flow Cytometry , Humans , Immunoglobulin E/biosynthesis , Lung/pathology , Mice , Mice, Inbred C57BL , Ovalbumin/immunology , Respiratory Hypersensitivity/pathology , Spleen/cytology , Spleen/immunology , T-Lymphocyte Subsets/immunology , Virus ReplicationABSTRACT
Fatal diabetic metabolic derangement is difficult to diagnose postmortem because of the paucity of characteristic morphologic findings. Hyperglycemia is an indicator of diabetic derangement. Conventional biochemical parameters for postmortem diagnosis of antemortem hyperglycemic states are not sufficiently resistant to antemortem and postmortem non-diabetic influences or are suited only for long and medium-term assessment of diabetes control. In the search for other, more reliable, indices of immediately antemortem blood glucose levels, we investigated the value of glycosylation levels of serum proteins with very brief biologic half-lives: a) In vitro studies were performed on the glycosylation course of the short-lived serum proteins alpha 1-antitrypsin (alpha 1-AT) and haptoglobin (HP). b) Glycosylation levels were measured after purification of alpha 1-AT and HP from sera of living and deceased non-diabetics and diabetics. c) The resistance of alpha 1-AT and HP glycosylation levels to autolysis was investigated. Our studies revealed the following: 1) alpha 1-AT and HP glycosylate considerably more rapidly than either albumin or hemoglobin. This rapid glycosylation, combined with the rapid turnover of both proteins; facilitates detection of short-term changes in glycemia. 2) alpha 1-AT and HP glycosylation levels are autolysis-stable and can be assessed even after advanced hemolysis. 3) alpha 1-AT and HP glycosylation levels appear to allow reliable ante- and postmortem discrimination between normoglycemic and hyperglycemic metabolic states. As a tool in the postmortem diagnosis of antemortem hyperglycemic states, alpha 1-AT and HP glycosylation levels combine the advantages of a short-term parameter with resistance to non-diabetic influences.
Subject(s)
Diabetes Mellitus/diagnosis , Haptoglobins/metabolism , Hyperglycemia/diagnosis , alpha 1-Antitrypsin/metabolism , Aged , Albumins/metabolism , Diabetes Mellitus/metabolism , Diabetes Mellitus/mortality , Female , Forensic Medicine/methods , Glycosylation , Humans , Hyperglycemia/blood , Male , Middle AgedABSTRACT
Age at death determination based on aspartic acid racemization in dentin has been applied successfully in forensic odontology for several years now. An age-dependent accumulation of D-aspartic acid has also recently been demonstrated in bone osteocalcin, one of the most abundant noncollagenous proteins of the organic bone matrix. Evaluation of these initial data on in vivo racemization of aspartic acid in bone osteocalcin was taken a step further. After purification of osteocalcin from 53 skull bone specimens, the extent of aspartic acid racemization in this peptide was determined. The D-aspartic acid content of purified bone osteocalcin exhibited a very close relationship to age at death. This confirmed identification of bone osteocalcin as a permanent, 'aging' peptide of the organic bone matrix. Its D-aspartic acid content may be used as a measure of its age and hence that of the entire organism. The new biochemical approach to determination of age at death by analyzing bone is complex and demanding from a methodologic point of view, but appears to be superior in precision and reproducibility to most other methods applicable to bone.
Subject(s)
Age Determination by Skeleton/methods , Aspartic Acid/analysis , Bone Matrix/chemistry , Osteocalcin/chemistry , Adult , Aged , Aged, 80 and over , Aspartic Acid/chemistry , Chromatography, High Pressure Liquid , Enzyme-Linked Immunosorbent Assay , Female , Forensic Anthropology , Humans , Male , Middle Aged , StereoisomerismABSTRACT
Determination of age at death on the basis of aspartic acid racemization in dentin is one of the most reproducible and accurate methods. In Germany, age estimation by this method has so far generally not been applied to living persons, since the extraction of a tooth exclusively for age estimation when it is not medically indicated is regarded as ethically and legally problematic. The development of a biopsy technique applicable to dentin took place against this background. Testing the technique and analysis of dentinal biopsy specimens revealed that the biopsy technique is a low-risk procedure that causes only minor discomfort to the affected person. It is readily practicable and facilitates standardized specimen removal. The relationship between the extent of aspartic acid racemization in dentinal biopsy specimens and age is very close, facilitating age estimation. A prerequisite for accurate results is the performance of biopsies under strictly standardized conditions. If this is guaranteed, age determination on the basis of aspartic acid racemization in dentinal biopsy specimens appears to be superior in precision to most other methods in living persons and can be used for all age groups.
Subject(s)
Aging/physiology , Aspartic Acid/chemistry , Dentin/chemistry , Forensic Dentistry/methods , Adolescent , Adult , Aged , Biopsy/methods , Child , Germany , Humans , Middle Aged , Reproducibility of ResultsABSTRACT
Age at death determination based on the extent of aspartic acid racemization in dentin has been reported to be highly accurate and reproducible. To test the applicability of this method to human bone, aspartic acid racemization in noncollagenous proteins of bone was investigated. A close relationship was found between age at death and the extent of aspartic acid racemization in osteocalcin, the most abundant noncollagenous protein of the organic bone matrix. Our findings indicate that osteocalcin is a permanent, 'aging' constituent of the organic bone matrix whose D-aspartic acid content increases with age because of in vivo racemization. Thus, the extent of aspartic acid racemization in bone osteocalcin is a measure of the age of the peptide and hence of the entire organism. The relationship between age at death and the extent of aspartic acid racemization in purified bone osteocalcin appears to be close enough to serve as a basis for determination of age at death in forensic medicine.
Subject(s)
Aspartic Acid , Bone and Bones/chemistry , Death , Age Determination by Skeleton , Age Factors , Forensic Medicine , Humans , Osteocalcin/chemistry , StereoisomerismABSTRACT
'Brain death' appears to be almost a closed chapter in medical history. But it remains fascinating, particularly regarding future developments, how medical discoveries, moral dilemmas, ethics and law have merged and lead to a common consent on the principal medical, ethical and juristic positions. After a consideration of these questions, topical problems concerning the extent and the limits of medical responsibilities, especially with regard to euthanasia and the problems of a broad interpretation of brain death are discussed.
Subject(s)
Brain Death , Ethics, Medical , Jurisprudence , Euthanasia , Germany , Humans , ResuscitationABSTRACT
We investigated whether measurement of aspartic acid racemization in intervertebral discs (IVD) could be used in the postmortem estimation of age at death. The extent of aspartic acid racemization in IVD tissues was found to increase with age. The rate of racemization turned out to be much higher in the nucleus pulposus than in the annulus fibrosus. The relation between age and the D-aspartic acid content in the anterior peripheral annulus fibrosus of IVD was close enough to allow postmortem estimation of age at death based on the extent of aspartic acid racemization in this tissue.