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Biomedical subjects

B Brinkmann

Publications and source records attributed to B Brinkmann.

At least 19 recordsLinked to original sources

Immunoreactivity enhancement with chelators for increasing the detection sensitivity of human PrPSc by Western blotting.

Prion diseases are neurodegenerative disorders affecting humans as Creutzfeldt-Jakob disease. The host-encoded prion protein (PrP(C)) will be converted into a structurally altered isoform (PrP(Sc)). PrP(Sc) differ in sizes and glycoform patterns and can be identified using molecular typing with Western blotting. The electrophoretic mobility of PrP(Sc) changes on treatment with metal ions or chelators prior to digestion with proteases. The effects of chelators applied to PrP(Sc) after protease digestion had not been examined in detail, we investigated these effects in this study. Application of EDTA, NTA and DTPA, and to a lesser extent EGTA, significantly enhanced PrP(Sc) signals in immunoblots. PrP(Sc) intensities increased two- to three-fold compared with untreated PrP(Sc). Since the immunoblot method is highly specific, sensitivity is the limiting factor. Enhancing sensitivity might be important in the determination of PrP(Sc) at levels close to or just below the limits of detection. It is to be expected that application of chelators to digested protein samples will increase the sensitivity of PrP(Sc) detection using the Western blot technique.

Animals↗

Do risk factors differ between explained sudden unexpected death in infancy and sudden infant death syndrome?

BACKGROUND: In Germany, 2910 infants died in 2004; for many infants the reason was clear, especially prematurity or congenital abnormalities. However, 394 babies die every year suddenly and unexpectedly. The cause may be immediately clear, but is often not obvious. AIMS: (1) To describe the causes of explained sudden unexpected death in infancy (SUDI) and (2) to compare risk factors for sudden infant death syndrome (SIDS) and explained SUDI. METHODS: A 3-year population-based case-control study in Germany, 1998-2001. RESULTS: 455 deaths, of which 51 (11.2%) were explained. Most of these deaths were due to respiratory or generalised infections. The risk factors for SIDS and explained SUDI were remarkably similar except for sleep position and breast feeding. Prone sleeping position is a major risk factor for SIDS (adjusted odds ratio (OR) 7.16, 95% confidence interval (CI) 3.85 to 13.31) but not for explained SUDI (adjusted OR 1.71, 95% CI 0.25 to 11.57). Not being breast fed in the first 2 weeks of life is a risk factor for SIDS (adjusted OR 2.37, 95% CI 1.46 to 3.84) but not for explained SUDI (adjusted OR 0.39, 95% CI 0.08 to 1.83). CONCLUSIONS: Prone sleeping position is a unique risk factor for SIDS. Socioeconomic disadvantage and maternal smoking are risk factors for both SIDS and explained SUDI, and provide an opportunity for targeted intervention.

Age Distribution↗

Sudden infant death syndrome: no increased risk after immunisation.

BACKGROUND: Although previous studies have shown either no association between immunisation and SIDS or even a decreased risk of SIDS, adverse effects, including death, from immunisations continue to cause concern, especially when a new vaccine is introduced. METHODS: A large case control study with immunisation data on 307 SIDS cases and 971 controls. RESULTS: SIDS cases were immunised less frequently and later than controls. Furthermore there was no increased risk of SIDS in the 14 days following immunisation. There was no evidence to suggest the recently introduced hexavalent vaccines were associated with an increased risk of SIDS. CONCLUSIONS: This study provides further support that immunisations may reduce the risk of SIDS.

Humans↗

Quantitative determination of taxine B in body fluids by LC-MS-MS.

A new specific and sensitive LC-MS-MS method for the detection of taxine B and isotaxine B, the main toxic pseudo-alkaloids from yew (Taxus sp.), in biological samples (blood, urine, gastric content) was developed. Biological samples were prepared for LC-MS-MS by means of solid-phase extraction (SPE) procedure and yielded a recovery of 86%. Chromatographic separation was achieved using an RP(18) column. Detection of taxine B and isotaxine B was performed using multiple reaction monitoring with m/z 584.2 as precursor ion, i.e. [M+H](+), of both isomers and m/z 194.3 and m/z 107.1 as product ions after collision-induced dissociation. Docetaxel was applied as internal standard. The method was fully validated for the analysis of blood samples. Linearity was proven in the range from 0.1-500 ng/g. The limit of detection and the limit of quantitation are 0.4 and 2 ng/g, respectively. The method was applied to the determination of taxine B and isotaxine B in four fatal cases (two humans, two horses) with suspected yew intoxication. Blood levels were 105, 168, 174 and 212 ng/g.

Adult↗

Coincidence of different structures of mucosa-associated lymphoid tissue (MALT) in the respiratory tract of children: no indications for enhanced mucosal immunostimulation in sudden infant death syndrome (SIDS).

Mucosa-associated lymphoid tissue (MALT) is the principal inductive site for mucosal immune responses that are capable of T and B cell responses and antigen-specific responses. In previous independent studies different structures of MALT, e.g. bronchus-, larynx- and nose-associated lymphoid tissue (BALT, LALT, NALT) have been described separately in various frequencies in the human respiratory tract over life spans. Because upper respiratory tract infections are common in infants, dysregulations of mucosal immune responses might be seriously involved in the aetiology of sudden infant death syndrome (SIDS). In the present study the coincidental occurrence of the three different MALT structures in the respiratory tract within the same patients were studied, and cases of SIDS and children who had died from different traumatic and natural causes of death (non-SIDS) were compared. First, the frequency of BALT and LALT in 46 children (35 SIDS, 11 non-SIDS) with or without NALT were examined. A tendency was found of a coincidence of respiratory MALT structures. In 50 additional cases of infant death (30 SIDS, 20 non-SIDS) from the multi-centric German Study on Sudden Infant Death Syndrome (GeSID) where death had occurred in the first year of life, the coincidence was evaluated. A coincidental occurrence of BALT, LALT and NALT or BALT and LALT (each about 30%) was found in both groups, whereby the coincidence in SIDS and the control patients did not differ. Interestingly, the children with coincidental MALT were strikingly older, supporting the hypothesis of respiratory MALT formation via environmental stimulation over time.

Bronchi↗

Sudden death after isobutane sniffing: a report of two forensic cases.

The intentional inhalation of a volatile substance ("sniffing") causing euphoria and hallucinations is an under-recognised form of substance abuse in children and adolescents with a high morbidity and mortality. Sudden death can be caused by cardiac arrhythmia, asphyxia or trauma. Two fatal cases of isobutane sniffing of cigarette lighter refill containing isobutane are reported. Toxicological investigations revealed the presence of isobutane in the heart blood and brain tissue of both cases (case 1: heart blood 0.1 microg/g, brain tissue 2.3 microg/g; case 2: heart blood 4.6 microg/g, brain tissue 17.4 microg/g) and the presence of its metabolite 2-methyl-2-propanol in the heart blood of both cases (0.5 and 1.8 microg/g, respectively). The histological investigations of the inner organs showed similar results in both victims. Autopsy findings, results of the histological and immunohistochemical investigations, toxicological findings and analytical procedures are discussed.

Administration, Inhalation↗

Mutations or exclusion: an unusual case in paternity testing.

In an immigration case with the scope of family reunification, the DNA extracted from the saliva samples of the male child, the alleged mother and the putative father was typed with 22 autosomal short tandem repeat (STR) systems. In seven STR systems, the alleged mother could be excluded from maternity, and the case then had to be regarded as a deficiency case. Taking this fact into consideration, only two exclusions were found for the putative father, and the question arose whether there was an exclusion of the putative father or the existence of two mutations. Autosomal STR typing could not clarify the case, but the application of eight Y-chromosomal markers showed that the alleged father could be excluded from paternity.

Child↗

Short amplicon STR multiplex for stain typing.

We developed a short tandem repeat (STR) typing kit based on DNA database systems that are included in, for example, the Interpol Standard Set of Loci recommendations (i.e., TH01, VWA, D3S1358, FGA) and the gender typing system Amelogenin. Two different multiplex sets were tested using the fluorescent dyes FAM, JOE, and VIC. The PCR results were compared to the commercially available AmpFISTR Blue kit, which contains the STRs D3S1358, VWA, and FGA. The advantage of our multiplex compared with the Blue kit was the generation of shorter amplicons (<200 bp) and the higher combined power of discrimination.

Amelogenin↗

An unusual case of homicidal near drowning followed by electrocution.

A young woman was found dead in a bath-tub with her facial orifices under water and current burn marks at the right big toe and at the neck near the hairline. Initial police investigations assumed an accidental electric shock due to a radio falling into the water. The forensic autopsy was followed by extensive histology and immunohistochemistry of the internal organs and skin marks. Intensive vital reactions of a near-drowning were detected that suggested a survival period of at least 20 min. Furthermore, an energy dispersive x-ray spectroscopy analysis on the current burn marks and on possible current applicators was performed, as well as reconstruction experiments with a cadaver of a pig that had been slaughtered shortly before. The results of all investigations revealed a two-step procedure with a primary near-drowning and a final current application with consecutive cardiac arrest.

Adult↗

The San Diego definition of SIDS: practical application and comparison with the GeSID classification.

The new definition of the term sudden infant death syndrome (SIDS) and the criteria introduced in San Diego for the subclassification of cases have been used to re-classify the first 100 consecutive cases of sudden and unexpected infant deaths that were registered with the German SIDS study (GeSID). Although there are 30 different variables that have to be considered in the general and stratified sections of the San Diego definition, it is practical, in particular, as an international standard to perform scientific studies. The comparison of the San Diego definition and the classification used for GeSID shows similarities in the methods but differences in the criteria used. Nevertheless, the numbers of cases classified as SIDS and borderline SIDS are similar (San Diego n=69, GeSID n=74). The SIDS IA criteria of the San Diego definition were not fulfilled by any case because metabolic screening and vitreous chemistry were not included in the GeSID investigation scheme. An important advantage of the San Diego definition is the introduction of the category of unclassified sudden infant death, which includes cases for which no autopsy was performed. This demonstrates that such cases cannot be classified as SIDS. In conclusion, we recommend the universal acceptance and use of the San Diego SIDS definition.

Age Distribution↗

Sudden death in childhood due to varicella pneumonia: a forensic case report with clinical implications.

Varicella (chickenpox) is one of the most frequent highly infectious diseases in childhood. It is caused by varicella-zoster virus. Lethal complications are rare. Focused on histological findings, we present a case of a sudden unexpected death of an otherwise healthy 18-month-old girl due to varicella-induced pneumonia. The histological and immunohistochemical investigations of the lung tissue revealed typical findings of a varicella pneumonia: haemorrhagic and necrotic nodules, intra-alveolar fibrin, numerous neutrophilic granulocytes, lymphocytes, plasmacells, macrophages, multinucleated giant cells and hyaline membranes. Varicella-related deaths are preventable by vaccine. To prevent complications and lethal outcome of varicella as reported here, the recommendations concerning vaccination against varicella must be taken into account in paediatric practice.

Adolescent↗

Unnatural causes of sudden unexpected deaths initially thought to be sudden infant death syndrome.

The aim of this clinicopathological study was to determine the frequency of infant deaths due to unnatural causes among cases of sudden and unexpected infant death. Nine institutes of legal medicine in Germany that took part in the German study on Sudden Infant Death Syndrome (GeSID), representing 35% of the German territory, investigated in a 3-year period (from 1998 to 2001) 339 cases of infant death that were not expected to be due to unnatural causes from the first external examination. All cases were investigated by complete, standardised, post-mortem examination including death scene investigation, autopsy, histology, toxicology and neuropathology. The frequency of unnatural deaths was 5.0% (n=17). The causes of death were head injury (n=7), suffocation (n=5), poisoning (n=2), neglect (n=2) and septicaemia due to aspiration of a foreign body (n=1). Two deaths were unsuspected accidents and 12 were due to infanticide. In 3 cases, it was not possible to differentiate between accidental death and infanticide. A complete postmortem examination including an analysis of the clinical history, death scene investigation, autopsy, histology, toxicology, and neuropathology is mandatory to differentiate sudden and unexpected deaths due to natural causes (e.g. SIDS) and cases of unnatural death.

Autopsy↗

HPLC with laser-induced native fluorescence detection for morphine and morphine glucuronides from blood after immunoaffinity extraction.

A new immunoaffinity solid phase extraction of morphine and its phase II metabolites, morphine-3-beta-D-glucuronide and morphine-6-beta-D-glucuronide is described. An immunoadsorber was applied which was created for the first time by the immobilisation of specific antibodies (polyclonal, host: rabbit) by the sol-gel method. The extraction method in combination with high performance liquid chromatography-fluorescence determination has been validated and shown to be applicable to blood samples of heroin victims in a low concentration range. Blood extracts were essentially free of interfering matrix components when compared to C8-extracts. Additionally, a novel, sensitive and selective detection system for wavelength-resolved analysis of laser-induced fluorescence coupled to HPLC was developed. The analytes were excited with a frequency tripled Ti:Sa laser (lambda=244 nm quasi cw). The total emission spectrum was recorded with a detection system consisting of an imaging spectrograph and a back-illuminated CCD camera. This technique of detection, combined with an extended optical path (at least 6 mm could be illuminated by the laser), resulted in an optimal fluorescence intensity of the analytes. The method permitted the analysis of morphine, morphine-3-beta-D-glucuronide and morphine-6-beta-D-glucuronide in a low concentration range and could be applied to a complex matrix such as postmortem blood samples because analyte peaks could be discriminated from matrix peaks by their characteristic emission spectra.

Chromatography, High Pressure Liquid↗

Infection, health problems, and health care utilisation, and the risk of sudden infant death syndrome.

AIM: To examine whether symptoms suggestive of infection, health problems, and health care utilisation are risk factors for SIDS. METHODS: Matched case-control study with 333 SIDS infants and 998 control infants matched for region, age, gender, and reference sleep. Information was obtained by parental interview, paediatrician completed questionnaire, and hospital admission data. RESULTS: No symptoms were associated with SIDS after adjustment for potential confounders. Illness in the last four weeks as reported by the paediatrician did not differ between cases and controls. Developmental problems and special investigations at any stage of life significantly increased the risk of SIDS (adjusted OR = 2.14 and 2.07). Admission to hospital after the first week of life was associated with an increased risk of SIDS (adjusted OR = 1.88). CONCLUSION: Symptoms of infection and illness are no longer risk factors for SIDS in communities such as Germany where few infants sleep prone. The increased risk of SIDS with developmental problems may indicate that infants which subsequently die of SIDS are abnormal or in some way vulnerable.

Analysis of Variance↗

Suffocation and poisoning--the hard-hitting side of Munchausen syndrome by proxy.

Munchausen syndrome by proxy (MSBP) is a severe and difficult to diagnose form of child abuse characterised by the simulation, aggravation or production of symptoms of illness in a child by an adult. MSBP often leads to multiple hospitalisations and has a high mortality and long-term morbidity. This study describes the cases of 5 families with 8 children affected who presented with unexplained neurological or gastrointestinal symptoms or even loss of consciousness. All were victims of poisoning or suffocation by their mothers. Two of those children died and were initially diagnosed as SIDS or natural death, respectively. They were only recognised as MSBP victims after another sibling had fallen ill with similar symptoms. The cases are discussed in consideration of the relevant literature. In addition warning signs of this forensically relevant syndrome and a strategy for the management of suspected MSBP cases are described.

Asphyxia↗

Backstabbing--a report of an unusual case.

A case of a 25-year-old male with a deep knife stab injury in the back is reported. The stab wound penetrated the left thorax and the left lung was injured, a thoracotomy was performed and the patient survived. The injured man could not remember what had happened, his bag was missing and the incident was therefore considered to be due to a robbery. Further results of the police investigations and the forensic pathology inspection revealed an extremely unusual accident constellation.

Accidents↗

Quantification of secondary dentine formation from orthopantomograms--a contribution to forensic age estimation methods in adults.

The accuracy of most age determination methods in adults is unsatisfactory and therefore more precise methods are required. Teeth provide several useful points about an individuals age. With advancing age secondary dentine is deposited along the wall of the dental pulp chamber leading to a reduction in the size of the pulp cavity. These age-related changes can be determined and measured from dental radiographs. The aim of this study was to explore if measurements of the size of the pulp cavity performed on orthopantomograms can be used for individual age estimation. Measurements were made digitally for 6 types of teeth from orthopantomograms of 168 individuals aged between 14 and 81 years old. To compensate for errors due to the technique of radiography-for instance differences in angulation-ratios of distances were calculated: pulp/root length, pulp/tooth length and pulp/root width at three different root levels. The width ratios of the pulp cavity showed significant correlation to the chronological age and the coefficient of determination (r(2)) was highest in the upper lateral incisors (r(2)=0.913) when an exponential or a logistic regression model was constructed. At the same distance with a linear regression model the coefficient of determination (r(2)) reached 0.839.

Adolescent↗

Development of a psilocin immunoassay for serum and blood samples.

After the immunisation of rabbits with a psilocin-specific immunogen, polyclonal antisera were obtained. With these antisera a competitive, heterogeneous radioimmunoassay for the detection of psilocin was developed. As tracer a derivative of psilocin was synthesised, which contained a tritiated CH(3) group. The antisera showed a specific reaction with psilocin. The cross-reactivity of structurally related endogenous substances like serotonin, tryptophan and tyrosine was below 0.01%. Also common drugs of abuse (Delta(9)-tetrahydrocannabinol, cocaine, morphine, amphetamine) showed negligible cross-reactivity (0.01-2%). Only tricyclic neuroleptics with a (dimethylamino)ethyl side-chain showed some cross-reactivity (20%). Spiked serum and blood samples were analysed with this new immunoassay and the results obtained were compared with the values measured with a validated GC-MS method.

Animals↗