Analysis of odontoblast gene expression using a novel approach,laser capture microdissection

Studying the mechanisms of molecular interactions in developing tissues demands sensitive molecular biological in vivo and in vitro techniques. Laser capture microdissection (LCM) allows for the isolation of mRNA in histological sections even from single cells, thus enabling the identification of in vivo gene expression products in closely circumscribed tissue areas. The aims of this study were to assess the optimal fixation, processing, and staining conditions to retrieve RNA from microdissected odontoblasts. Fluorometric assays and RT-PCR analysis of alpha 1(I) collagen, dentin sialophosphoprotein (Dspp), and osteocalcin (OC) confirmed that the total RNA isolated from day 0 and day 3 captured odontoblasts was sufficient in quantity and quality. Our results indicate that individual odontoblasts obtained by LCM are morphologically intact and chemically unaltered, allowing accurate molecular and biochemical analyses.     

Large-field S-cone flicker test

Background: Disturbances of blue color vision and of temporal contrast sensitivity can indicate early damage in glaucoma. For the present study a quick and easy test was devised which examines both functions ai one time by testing the temporal contrast sensitivity of a blue flickering light on an intense yellow background. Methods: Large coextensive background and test fields (85°) are used, making fixation uncritical. Detailed experiments were made in two normal subjects to derive spectral sensitivity curves from flicker-fusion frequency (FFF) versus intensity functions and to obtain complete temporal contrast-sensitivity (De Lange) curves under different levels of adaptation and test lights. After selection of appropriate luminances and one stimulation frequency from these experiments, test-retest variability was studied in four subjects in five repetitions. In addition, normal values were collected from 22 subjects. Results: Spectral sensitivities for two levels of FFF (15 Hz and 44 Hz) agree with Stiles' ₁ at the low and with ₄ at the high FFF. Temporal contrast-sensitivity curves show a low-frequency section with peak sensitivity at 1 Hz and a high-frequency section with a peak at around 4 Hz. From the basic experiments the following conditions for the clinical examination were selected: Background luminance 2600 cd/m², test luminance at 451 nm 0.8 cd/m², stimulation frequency 4 Hz. The test-retest variability showed an acceptable intraclass correlation coefficient (0.6). Conclusions: The present experiments carried out with a very large stimulus led to meaningful results which are in rather good agreement with results reported in the literature on small-field stimuli. The blue-on-yellow flicker test carried out under the conditions mentioned above is a quick and easy test which could be helpful in improving early glaucoma diagnosis.     

Hippocampal loss of the GABAA receptor α1 subunit in patients with chronic pharmacoresistant epilepsies

Alterations of gamma aminobutyric acid (GABA)-mediated neurotransmission have been implicated in the pathogenesis of epilepsies. Here we examine the distribution of the GABA_A receptor in the hippocampus of 78 surgical specimens from patients with chronic pharmacoresistant focal epilepsies. The receptor was localized immunohistochemically with the monoclonal antibody bd-24 which selectively recognizes the ₁ subunit of the GABA_A receptor. The results were compared with the receptor distribution of 28 normal hippocampal specimens obtained at autopsy. In the great majority of the surgical specimens a loss of GABA_A receptor immunoreactivity was present in CA1 (92.3%), CA4 (78.2%), the dentate granular cell layer (70.5%) and the molecular layer of the dentate gyrus (65.4%). The subiculum revealed a normal staining pattern in all but 4 cases. In no instance did we observe an increase of immunoreactivity in any region or cell population. The decrease of GABA_A receptor immunoreactivity was closely related to neuronal loss in the respective specimen and to Ammon's horn sclerosis. There was no correlation between GABA_A receptor loss and the patient's age at surgery, duration of seizures, age at onset of seizures and to the presence or absence of secondary generalized tonic clonic seizures. The data suggest that the observed loss of GABA_A receptor immunoreactivity is a secondary phenomenon rather than an event that is relevant for the pathogenesis of epileptic seizures.     

The effect of accident mechanisms and initial findings on the long-term course of whiplash injury

The aim of this study was to assess the relationships between accident mechanisms as well as initial findings and the long-term course of whiplash injury. A representative sample of 117 consecutive patients referred by primary care physicians was followed-up over 12 months. Fractures or dislocations of the cervical spine, head trauma and pre-existing neurological disorders were exclusion criteria. The interval between the accident and the baseline examination was 7.4 days (SD 4.2 days). Assessment included accident features (e.g. passenger position in the car, head restraint, head position, type of collision), initial symptoms (e.g. intensity and onset of pain, symptoms of neurological dysfunction, multiple symptom score), and signs (restricted neck movement, neurological deficits). At the 1-year examination, patients were divided into an asymptomatic and a symptomatic group and were compared with respect to accident features and baseline findings. Twenty-four percent of patients were still symptomatic after 1 year. Analysing accident mechanisms separately, rotated or inclined head position was the primary feature related to symptom persistence (P=0.005). The symptomatic group scored higher at baseline on the multiple symptom rating (P=0.004) and had a higher incidence of initial headache (P=0.004) and neurological symptoms (P=0.008) together with a higher intensity of headache (P=0.0002) and neck pain (P=0.0009). The following set of initial variables predicted persistence of symptoms at 1 year (logistic regression): intensity of neck pain (P=0.001) and headache (P=0.009), rotated or inclined head position (P=0.02), unpreparedness at the time of impact (P=0.01) and car stationary when hit (P=0.01). In conclusion, accident mechanisms and initial findings suggestive of more severe injury were significantly related to long-term persistence of symptoms after whiplash injury.This study was supported by the Swiss National Science Foundation (project number: 3.883-0.88) and the Swiss Accident Insurance Company (Schweizerische Unfallversicherungsanstalt), Berne     

Urinary excretion of adenosine deaminase binding protein in neonates treated with tobramycin

The potential tubulotoxicity of tobramycin and cefotaxim were assessed in neonates by measuring the urinary level of adenosine deaminase binding protein (ABP) and urinary ₁-microglobulin and ₂-microglobulin. In a prospective study, 33 neonates who received tobramycin and cefotaxim for suspected neonatal sepsis were compared with 48 untreated newborns during the first 10 days of life. The urinary concentrations of ABP and its excretion rates, corrected for body weight and body surface area, were significantly increased from the 1st day of treatment. Urinary ₁-microglobulin and ₂-microglobulin were not elevated under tobramycin and cefotaxim during the first 2 days of treatment. We conclude that ABP may be a sensitive marker for the detection of proximal renal tubular injury during tobramycin and cefotaxim treatments of neonates. The increase in urinary ABP which occurs before an elevation of urinary ₁-microglobulin and ₂-microglobulin may reflect earlier structural than functional alterations. However, since none of the treated infants had signs of electrolyte disorders or glomerular dysfunction, the clinical relevance of ABP measurement should be reevaluated.     

Alcohol consumption and gender in the 20th century: the case of Switzerland

Given the changes of gender roles in this century it is hardly justified to assume constant proportions of alcohol consumption for males and females. The purpose of the study was to reconstruct the consumption trends of males and females in Switzerland since the beginning of the 20th century. Cirrhosis mortality and survey data were used to disaggregate by sex the per capita alcohol consumption based on liver cirrhosis mortality suggests that the evolution of alcohol consumption in Switzerland followed a parallel course for both sexes only until the 1930 s. The low consumption during World War II and the evident increase until the beginning of the '60s seem to have resulted above all from the variations in consumption of beer by men. The decrease in total alcohol consumption observed since the '70s is also most probably due only to men; there is no indication of a decreasing consumption by women. The tendency of male and female consumption patterns to become more similar should be taken into account in the prevention of alcohol misuse.     

Psychisch Kranke: Gesehen aus zwei verschiedenen Perspektiven Ein methodischer Beitrag zur Einstellungsforschung in der Psychiatrie

Zusammenfassung Anhand von Daten aus zwei Bevölkerungsumfragen zur Einstellung gegenüber psychisch Kranken wird untersucht, welchen Einfluss die Variation der Perspektive-die des Befragten selbst versus die der anderen-auf das Befragungsergebnis hat. Wie erwartet fällt das Urteil über psychisch Kranke positiver aus, wenn nach der eigenen Meinung gefragt wird, als wenn man das Urteil der anderen einschätzen lässt. Der Unterschied zwischen beiden Frageformulierungen nimmt mit dem Grad der Schulbildung zu und erreicht bei Personen mit Abitur ein substantiell bedeutsames Ausmass. Darüber hinaus sind die Antworten eindeutiger und pointierter, wenn nach der persönlichen Meinung gefragt wird.

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Summary The impact of the change in perspective (i.e. the personal opinion of those questioned versus what they perceive the others' opinion toward the mentally ill to be) on the results of the questionnaire is examined based on data from two population surveys measuring attitudes towards the mentally ill. As expected, the respondents' attitudes towards the mentally ill are more positive if they are asked to give their own opinion. The impact that the variation of the two formulations of these questions has on the respondents increases with their level of education and reaches a substantial amount among those with Abitur. Furthermore, the answers are clearer and more definite if those questioned are asked for their personal opinion.

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Résumé L'impact d'un changement de perspective (opinion personnelle comparée à l'opinion générale telle qu'elle est personnellement perçue) est examiné dans le cadre de deux études de population mesurant l'attitude envers les malades psychiques. Comme on pouvait s'y attendre, l'attitude des répondants est beaucoup plus positive lorsqu'il s'agit de leur opinion personnelle. La divergence entre l'opinion personnelle et la perception de l'opinion générale augmente au fur et à mesure que le niveau d'éducation s'accroît. Les réponses sont d'autre part bien plus clairement formulées lorsque les personnes experiment leur opinion personnelle.

     

Histochemical identification of endogenous lectins using labelled neoglycoproteins in human head-and-neck squamous cell carcinoma

According to the "Population-based cancer register" of the Federal Republic of Germany only malignant neoplasms of the buccal cavity, the pharynx and larynx as well as cancers of the respiratory tract show an increasing rate of incidence and mortality. The molecular mechanisms and etiological factors causing this phenomenon are still little understood despite intensive research work. Recognition between receptors on a cellular level may be mediated by specific amino acid sequences on the level of protein-protein recognition. Additionally, the interactions between cell sugars and the corresponding protein receptor may play a decisive role in development, regeneration and organisation of cells and tissue. The high specificity of the binding of biotinylated neoglycoproteins in tissue sections enables to detect glycohistochemically binding sites for the carbohydrate ligands of the glycosylated carrier protein. The evidence of lectins in squamous cell cancer of the oral cavity, oropharynx, larynx, and hypopharynx has not been established so far. Squamous cell cancer tissue samples of twelve patients with different tumour locations were investigated by incubation of sections of paraffin-embedded samples and application of an avidin-biotin-peroxidase complex for visualisation with synthetic biotinylated neoglycoproteins. Altogether 168 stained sections were evaluated including controls. Pronounced cytoplasmatic staining was seen with the following neoglycoproteins: sialic acid-bovine serum albumin (BSA), glucuronic acid-BSA, N-acetylglucosamine (glcNAc)-BSA, N-acetylgalactosamine (beta-galNAc)-BSA, lactose-BSA, maltose-BSA, mannose-BSA, mannose-6-phosphate-BSA. No corresponding lectins seems to exist for the following investigated sugars: fucoidan, heparin, and the alpha-anomeric form of N-acetylgalactosamine, because no specific staining was seen.(ABSTRACT TRUNCATED AT 250 WORDS)     

Familiale Beziehungsmuster und Patientenkarriere männlicher schizophrener Kranker

Zusammenfassung Mit Hilfe eines Persönlichkeitsfragebogens (Gießen-Test), der von beiden Eltern und dem schizophrenen Kranken bei dessen Entlassung aus der ersten stationären psychiatrischen Behandlung ausgefüllt worden war, wurde die Personenwahrnehmung und damit mittelbar die Beziehung in der Familie untersucht. Über alle drei Selbstbilder und sechs Fremdbilder von insgesamt 28 Familien mit einem schizophrenen Sohn sowie von 29 als Vergleichsgruppe dienenden Familien mit einem chirurgisch kranken Sohn wurde eine Clusteranalyse berechnet, die die Abgrenzung von fünf Clustern erlaubte, von denen zwei vor allem durch Familien mit Schizophrenen und zwei vor allem durch solche mit organisch kranken Söhnen gebildet wurden; ein Cluster war unspezifisch. Ein 2-Jahres-follow-up ergab, daß 13 schizophrene Kranke erneut die Klinik aufsuchen mußten. Es fand sich keine statistisch bedeutsame Beziehung zwischen den untersuchten psychiatrischen und soziodemographischen Parametern und dem Rehospitalisierungsrisiko. Als besonders gefährdet erwiesen sich jedoch Familien, in denen sich nach den Ergebnissen der Clusteranalyse ein Konflikt zwischen Eltern und Sohn um die Definition als Kranker auf dem Hintergrund rigider Normvorstellungen abbildete. Die Übernahme der Krankenrolle durch den Patienten ging hingegen nur in der Hälfte der Fälle mit einer Rehospitalisierung einher, führte also nicht zwangsläufig zur Ausgrenzung aus dem Familienverband. Söhne aus Familien, die als besonders gesund imponierten, aber auch Söhne aus Familien mit dominanten Müttern wurden im Untersuchungszeitraum nicht erneut in die Klinik aufgenommen.

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Summary Patterns of personal relationships within the families of male schizophrenic patients, and their relationship to clinical course and outcome, were examined by means of a personality questionnaire (Gießen-Test), completed by both parents and by the schizophrenic patient himself at the time of discharge from his first psychiatric in-patient treatment. The test yields scores related to the self-perceptions of the individual and to his perceptions of the other nuclearfamily members. A cluster analysis was performed on all three self-perception scores and six scores of the perceptions the three family members had of each other, for a total of 28 families each with a schizophrenic son and for a control group of 29 families each with a surgically-ill son. This analysis allowed five clusters to be distinguished, two of which consisted predominantly of families with schizophrenic sons and two predominantly of families with surgically-ill sons. The remaining cluster comprised a mixed group. A two-year follow-up showed that 13 schizophrenic patients had been readmitted to hospital. There was no statistically significant relationship between the psychiatric and sociodemographic parameters examined and the re-hospitalization rate. Readmissions were, however, more frequent from families in which the research findings had indicated conflicting mutual perceptions of parents and sons, against a background of rigid notions of normality. Acceptance by the patient of the notion of being ill was associated with re-hospitalization in only about half of the cases, so that it had not necessarily led to rejection by the family unit. No patients were readmitted during the follow-up period from families which had represented themselves as particularly healthy, or from families with dominant mothers.

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Teil des von der Deutschen Forschungsgemeinschaft unterstützten Projektes Interaktion in Familien mit Schizophrenen