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.     

Chronic motor neuropathies: response to interferon-beta1a after failure of conventional therapies

OBJECTIVES: The effect of interferon-beta1a (INF-beta1a; Rebif) was studied in patients with chronic motor neuropathies not improving after conventional treatments such as immunoglobulins, steroids, cyclophosphamide or plasma exchange. METHODS: A prospective open study was performed with a duration of 6-12 months. Three patients with a multifocal motor neuropathy and one patient with a pure motor form of chronic inflammatory demyelinating polyneuropathy were enrolled. Three patients had anti-GM1 antibodies. Treatment consisted of subcutaneous injections of IBF-beta1a (6 MIU), three times a week. Primary outcome was assessed at the level of disability using the nine hole peg test, the 10 metres walking test, and the modified Rankin scale. Secondary outcome was measured at the impairment level using a slightly modified MRC sumscore. RESULTS: All patients showed a significant improvement on the modified MRC sumscore. The time required to walk 10 metres and to fulfil the nine hole peg test was also significantly reduced in the first 3 months in most patients. However, the translation of these results to functional improvement on the modified Rankin was only seen in two patients. There were no severe adverse events. Motor conduction blocks were partially restored in one patient only. Anti-GM1 antibody titres did not change. CONCLUSION: These findings indicate that severely affected patients with chronic motor neuropathies not responding to conventional therapies may improve when treated with INF-beta1a. From this study it is suggested that INF-beta1a should be administered in patients with chronic motor neuropathies for a period of up to 3 months before deciding to cease treatment. A controlled trial is necessary to confirm these findings.     

Depression and schizophrenia in epilepsy: social and biological risk factors

BACKGROUND: In a retrospective study we investigated the role of social and biological risk factors for the development of major depression and schizophreniform psychoses in epilepsy. We tested the hypotheses that social risk factors are associated with depression and biological risk factors are associated with schizophreniform psychoses. METHOD: We studied 25 patients with epilepsy and paranoid-hallucinatory psychosis, 25 patients with epilepsy and major depression, and 50 non-psychiatric epilepsy patients (controls) with respect to biological and psychosocial variables. RESULTS: Schizophrenic patients had an earlier age of onset of epilepsy and a more severe epilepsy as characterised by history of status epilepticus, multiple seizure types and severity of seizures compared to non-psychiatric controls. Simple seizure symptoms were often vegetative and EEGs showed various abnormalities including temporal lobe discharges but no lateralisation to either side. With respect to antiepileptic drugs (AED) there were only few significant differences between groups: Polytherapy as well as treatment with phenytoin (DPH) was more frequent in psychotic patients as compared to non-psychiatric patients. Patients with psychoses were also characterized by a disturbed familial background, lack of interpersonal relationships, social dependency and professional failure. Depressive patients were significantly older than non-psychiatric controls and they suffered more frequently from focal epilepsies arising from the temporal lobe. They did not differ from controls with respect to severity of epilepsy. Treatment with valproate (VPA) was inversely linked with depression, suggesting that VPA may have prophylactic antidepressive properties in epilepsy patients. There were no psychosocial variables significantly linked with depression. CONCLUSIONS: In this study, patients with different forms of psychiatric complications in epilepsy could clearly be distinguished from controls. However, we could not confirm the simple hypothesis that there are biological predictors for schizophreniform psychoses and psychosocial predictors for major depression. Neurological and sociological variables seem linked with both, suggesting a multifactorial etiology.     

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.