OPA1, encoding a dynamin-related GTPase, is mutated in autosomal dominant optic atrophy linked to chromosome 3q28.

Autosomal dominant optic atrophy (ADOA) is the most prevalent hereditary optic neuropathy resulting in progressive loss of visual acuity, centrocoecal scotoma and bilateral temporal atrophy of the optic nerve with an onset within the first two decades of life. The predominant locus for this disorder (OPA1; MIM 165500) has been mapped to a 1.4-cM interval on chromosome 3q28-q29 flanked by markers D3S3669 and D3S3562 (ref. 3). We established a PAC contig covering the entire OPA1 candidate region of approximately 1 Mb and a sequence skimming approach allowed us to identify a gene encoding a polypeptide of 960 amino acids with homology to dynamin-related GTPases. The gene comprises 28 coding exons and spans more than 40 kb of genomic sequence. Upon sequence analysis, we identified mutations in seven independent families with ADOA. The mutations include missense and nonsense alterations, deletions and insertions, which all segregate with the disease in these families. Because most mutations probably represent null alleles, dominant inheritance of the disease may result from haploinsufficiency of OPA1. OPA1 is widely expressed and is most abundant in the retina. The presence of consensus signal peptide sequences suggests that the product of the gene OPA1 is targeted to mitochondria and may exert its function in mitochondrial biogenesis and stabilization of mitochondrial membrane integrity.

Genomic rearrangements in OPA1 are frequent in patients with autosomal dominant optic atrophy.

INTRODUCTION: Autosomal dominant optic atrophy (ADOA) is considered as the most common form of hereditary optic neuropathy. Although genetic linkage studies point to the OPA1 locus on chromosome 3q28-q29 as by far the most common gene locus, previous screening studies-based on sequencing of the coding exons-detected OPA1 mutations in only 32-70% of ADOA patients. We therefore hypothesised that larger deletions or duplications that remained undetected in previous screening approaches may substantially contribute to the prevalence of OPA1 mutations in ADOA. METHODS: 42 independent ADOA patients were analysed for the presence of genomic rearrangements in OPA1 by means of multiplex ligation probe amplification (MLPA). Deletions or duplications were confirmed either by long distance polymerase chain reaction (PCR) and breakpoint sequencing or loss of heterozygosity analyses with flanking microsatellite markers. Patients underwent ophthalmological examination including visual acuity, colour vision testings, perimetry and funduscopy. RESULTS: We identified genomic rearrangements in 8 of 42 patients, including single exon deletions of exon 9 and exon 24, respectively, a deletion of exons 1-5, two different deletions of the complete OPA1 gene as well as a duplication of the exons 7-9, with the latter being present in three unrelated families. Patients' phenotypes were highly variable, similar to patients with point mutation in OPA1. DISCUSSION: Our findings show that gross genomic aberrations at the OPA1 gene locus are frequent in ADOA and substantially contribute to the spectrum and prevalence of OPA1 mutations in ADOA patients. They further strengthen the hypothesis that haploinsufficiency is a major pathomechanism in OPA1 associated ADOA.

[How to distinguish between autosomal dominant optic atrophy and Leber's hereditary optic neuropathy]. / Autosomal-dominant erbliche Optikusatrophie vs. Lebersche Optikusneuropathie. Wie kann man sie unterscheiden?

BACKGROUND: Patients with long-lasting bilateral optic atrophy showed typical clinical features of autosomal dominant optic atrophy (ADOA). Molecular genetic analysis identified them as atypical cases of Leber's hereditary optic neuropathy (LHON). METHOD: Three patients with bilateral optic atrophy and central scotomas of their visual fields were clinically diagnosed with ADOA. Samples of lymphocytic genomic DNA were amplified with polymerase chain reaction, and analysis of the coding exons including the flanking intron/UTR sequences of the OPA-1 gene was performed. However, no ADOA-associated mutations were found. We therefore analysed the total lymphocyte mitochondrial DNA for all common LHON mutations in these patients. RESULTS: Three patients from three unrelated pedigrees (two men, one woman) who were clinically diagnosed as suffering from ADOA did not harbor any typical mutation of the OPA-1 gene. However, analysis of their mitochondrial DNA showed that they harbored the 3460, 11778, and 14484 LHON mutations. The patients were identified as atypical cases of LHON. The pedigrees of the patients fulfilled the criteria for both dominant and mitochondrial-maternal transmission in all cases. The clinical picture of LHON differed remarkably from the classic course of LHON. CONCLUSIONS: To identify atypical LHON patients with bilateral optic atrophy and central scotomas in the visual field and to distinguish them from ADOA patients, careful molecular genetic analysis is necessary. In these rare cases, only double examinations of both the genomic and the mitochondrial DNA will allow these patients to be adequately advised.

How deeply does your mutant sleep? Probing arousal to better understand sleep defects in Drosophila.

The fruitfly, Drosophila melanogaster, has become a critical model system for investigating sleep functions. Most studies use duration of inactivity to measure sleep. However, a defining criterion for sleep is decreased behavioral responsiveness to stimuli. Here we introduce the Drosophila ARousal Tracking system (DART), an integrated platform for efficiently tracking and probing arousal levels in animals. This video-based platform delivers positional and locomotion data, behavioral responsiveness to stimuli, sleep intensity measures, and homeostatic regulation effects - all in one combined system. We show how insight into dynamically changing arousal thresholds is crucial for any sleep study in flies. We first find that arousal probing uncovers different sleep intensity profiles among related genetic background strains previously assumed to have equivalent sleep patterns. We then show how sleep duration and sleep intensity can be uncoupled, with distinct manipulations of dopamine function producing opposite effects on sleep duration but similar sleep intensity defects. We conclude by providing a multi-dimensional assessment of combined arousal and locomotion metrics in the mutant and background strains. Our approach opens the door for deeper insights into mechanisms of sleep regulation and provides a new method for investigating the role of different genetic manipulations in controlling sleep and arousal.

Segregation patterns and heteroplasmy prevalence in Leber's hereditary optic neuropathy.

PURPOSE: To investigate the segregation pattern of the mitochondrial DNA mutation at nucleotide position 3460 responsible for Leber's hereditary optic neuropathy (LHON) and to determine the prevalence of heteroplasmy for the three primary LHON mutations at positions 11778, 3460, and 14484. METHODS: Segregation analysis was performed in a cross-sectional study by determining the level of heteroplasmy in blood leukocytes of 23 LHON patients and unaffected carriers from four unrelated families. One family comprising two affected and three unaffected carriers was followed over 5.5 years for a longitudinal segregation analysis of heteroplasmy. The percentage of mutant mtDNA was determined using a novel procedure of fluorescence-based primer extension and restriction fragment length polymorphism analysis. The prevalence of heteroplasmy was assessed by determining the number of genealogically unrelated LHON pedigrees with heteroplasmic maternal family members from the LHON patient records of the Department of Ophthalmology, University of Tübingen, Germany. RESULTS: The authors observed a marked variability in the degree of heteroplasmy levels within each pedigree and a tendency toward a higher mutant allele frequency in offspring generations. Disease expression was correlated with higher levels of mutant mtDNA molecules. Longitudinal analysis revealed no statistically significant decrease in the heteroplasmy level in the family studied but a reduction of 11% and 12% in one affected and one unaffected individual, respectively. In 167 genealogically unrelated LHON families the prevalence of heteroplasmy was 5.6%, 40%, and 36.4% for the 11778, 3460, and 14484 LHON mutations, respectively. CONCLUSIONS: Cross-sectional studies of heteroplasmy for the 3460 LHON mutation suggest that the genotype shifts toward a higher mutational load in offspring generations. Long-term decrease in the blood mutant load in single cases indicates negative selection of the mutant allele in the hematopoietic cell system. The prevalence of heteroplasmy varies significantly between the different primary LHON mutations, suggesting genotypical differences in disease expression.

Relationship of properties of polycyclic aromatic hydrocarbons to sequestration in soil.

A study was conducted to determine whether the sequestration of 21 polycyclic aromatic hydrocarbons (PAHs) in soil was correlated with their properties. From 22 to 58% of the PAHs was not extracted with n-butanol after their addition to soil. After 28 days of aging, the percentage of the PAHs remaining in the soil increased to 47-77%; however, nearly all of each compound was recovered by Soxhlet extraction. Correlations were based on the amounts of aged compound extracted with butanol. Properties of compounds used in the correlations included Kow, molecular length and molecular-connectivity indices (MCIs). No one property, including log Kow, resulted in an R2 value greater than 0.26. A chain MCI (2 chi vCH) together with log Kow or a first-order MCI (1 chi) resulted in R2 values of 0.49 and 0.54, respectively. The data suggest that the properties tested are not important to predicting the sequestration of PAHs in soil.

Influence of soil moisture on the sequestration of organic compounds in soil.

A study was conducted as a part of continuing investigation of the effect of soil moisture on the sequestration of organic compounds aged in the soil. Here, experiments focused on the effects of moisture changes within the soil before, during, and after contaminant addition. The extractability of aged (68 d) phenanthrene was greater from soil that had been subjected to wetting and drying cycles prior to solute addition as compared to soil initially maintained at constant moisture. The recovery of phenanthrene added to moist soil was increased relative to extractability from soil that was air-dried at the time of the contaminant addition. Repeated wetting and drying of soil after the addition of atrazine or phenanthrene resulted in decreased extractability of the compounds as compared to samples maintained at constant moisture. A method for rapidly sequestering contaminants is proposed and may be useful in limiting the time required for laboratory studies involving "aged" contaminants. These data build upon the findings of earlier work from our laboratory and indicate that changes in the moisture conditions of soil can affect the availability of sequestered contaminants possibly through alterations in the structure of the natural solid.

[Leber optic neuropathy in women and children]. / Lebersche Optikusneuropathie (LHON) bei Frauen und Kindern.

BACKGROUND: Leber's hereditary optic neuropathy is associated with point mutations in the mitochondrial DNA (mtDNA) that appear to be pathogenetic for this disease. These mutations affect nucleotide positions 3460, 11778 and 14484. Does the clinical course of LHON differ between men, women and children? MATERIALS AND METHODS: We reviewed the clinical and molecular genetic characteristics of 15 visually symptomatic patients with the clinical diagnosis of LHON (11 women and 4 male children) and compared them with 66 men with LHON. RESULTS: LHON was confirmed clinically and with molecular genetic methods in all cases. Men, women and children showed no differences: Classic fundus findings and typical visual field defects were equally found in both sexes. However, age at the beginning of the disease, severity of LHON and rate of spontaneous recovery differed between groups. Women were older (19-55 years, average 31.3 years) than men (15-53 years, average 24.3 years) at the beginning of the disease. Women suffered more severely from LHON. Spontaneous recovery of vision in women was extremely rare. Many more women had a LHON-affected mother than men. All the affected children (9-14, average 11.7 years at the beginning of the disease) did not have a good visual outcome. CONCLUSIONS: There are some differences in the course of LHON between men and women, concerning age, severity of LHON and rate of spontaneous recovery. Children may also have an unfavorable prognosis.

[Leber optic neuropathy with clinical improvement]. / Lebersche Optikusneuropathie mit klinischer Besserung.

BACKGROUND: Spontaneous recovery in Leber's hereditary optic neuropathy is rare. Does the clinical course of Leber's hereditary optic neuropathy (LHON) differ between patients with and without spontaneous recovery? MATERIALS AND METHODS: We compared the clinical and molecular genetic characteristics of 12 visually symptomatic patients having the classical clinical course of LHON who recovered spontaneously with those of 60 who did not. RESULTS: Classical fundus findings and typical visual field defects were comparable in the two groups; vision improved within 18 months in all cases. The worst visual acuity during the acute stage of LHON was 0.03 in the recovery group. Patients with the 3460 and especially the 14484 mutation had a better chance of recovery. No patient with the 11778 mutation who recovered had secondary mutations. Among patients who recovered women were underrepresented and heteroplasmy was more common. Some families showed a raised rate of clinically affected members with recovery. CONCLUSIONS: The clinical picture of LHON remains the same regardless of whether the patient recovers spontaneously. A higher rate of spontaneous recovery characterizes some families. Spontaneous recovery is rare in women. Heteroplasmy is frequent in patients with recovery. Our results show a better clinical course of LHON in patients with the 11778 mutation without secondary mutations. Prognosis is better if the peripapillary microangiopathy is seen for a relatively long period, and there is only partial optic atrophy.

[Diagnostic error in Leber's optic neuropathy. Value of clinical and molecular genetic studies]. / Fehldiagnose Lebersche Optikusneuropathie (LHON). Stellenwert klinischer und molekulargenetischer Untersuchungen.

BACKGROUND: Leber's hereditary optic neuropathy is associated with point mutations in the mitochondrial DNA (mtDNA) that appear to be pathogenic for this disease. These mutations affect nucleotide positions 3460, 11,778 and 14,484. MATERIALS AND METHODS: We reviewed the clinical and molecular genetic characteristics of 29 visually symptomatic patients with the clinical diagnosis of LHON. RESULTS: Nine patients really suffered from LHON, but in 20 patients other ocular diseases could be proven. Degeneration of the retina and choroid was most common (seven patients), followed by vascular optic disease (six patients). Three patients suffered from tobacco-alcohol amblyopia, two from optic neuritis and two from autosomal dominant optic neuropathy. CONCLUSIONS: The clinical diagnosis of LHON is strengthened by a proven maternal inheritance and clinical signs such as a severe decrease in visual acuity, central or centrocecal scotomas in the perimetry and pseudoedema of the optic disc, followed by optic atrophy. Pathognomonic clinical signs of LHON are twisted vessels and ectatic capillaries in the fundus of these patients and their relatives of the maternal line, i.e., peripapillary microangiopathy. A careful analysis of the patients' pedigrees, anamnesis and the functional and morphological results of the clinical examinations helps to avoid misdiagnosis of the disease. However, the expensive and time-consuming molecular genetic analysis is always necessary to confirm or exclude the diagnosis of LHON.

[Correlation between clinical and molecular genetic findings in Leber's optic atrophy]. / Korrelation klinischer und molekulargenetischer Befunde bei Leberscher Optikusneuropathie (LHON).

Leber's hereditary optic neuropathy (LHON) is associated with point mutations of mitochondrial DNA (mtDNA) that appear to be pathogenetic for this disease. These mutations affect nucleotide positions 3460, 4160, 11,778, 14,484, and possibly 15,257. The pathogenetic significance of other mtDNA point mutations (secondary mutations) is less clear. We reviewed the clinical and molecular genetic characteristics of 29 visually symptomatic patients from 26 families. In addition, we studied 54 relatives of the maternal line of these patients. Sixteen of them underwent clinical and molecular genetic examination; 38 underwent only molecular genetic examination. The 29 affected individuals showed a male predominance of 93.1% (27/29) and ages of onset of visual loss ranging from 15 to 55 years. The time interval between affected eyes was never longer than 1 year. Tobacco and/or alcohol abuse was common. Peripapillary microangiopathy was found in 20.7% (6/29) of our patients. The number of patients with peripapillary microangiopathy seems to be small, but we could not examine all patients early after onset of the disease: the time of first examination is critical for the diagnosis of peripapillary microangiopathy. From the 16 relatives who underwent clinical examination, 62.5% (10/16) also had peripapillary microangiopathy. Eighteen patients were analyzed by brain computed tomography or magnetic resonance imaging. Four definitely pathological results seem remarkably high in comparison with the results of other authors. Our LHON patients and their relatives invariably had an identical pattern of point mutations, both primary as well as secondary. Of the LHON patients, 79.3% had the primary mutation at position 11,778, 20.7% at position 3460. Different numbers and combinations of secondary mutations were observed in a large portion of both groups. Three patients with the 11,778 mutation noticed remarkable visual recovery. There was no clear correlation between the type and number of point mutations in the individual and the severity of the disease.

[Leber's hereditary optic neuropathy]. / Lebersche Optikusneuropathie.

Leber's hereditary optic neuropathy (LHON) is a rare disease primarily affecting the retinal ganglion cells. In most cases patients with LHON develop permanent visual loss with a large central scotoma in the visual field of both eyes. The optic disc becomes partially or completely pale. At the onset of the disease many patients are considered to suffer from an optic neuritis and are treated under the diagnostic and therapeutic regimen of optic neuritis. LHON is mostly only considered when high dose cortisone therapy fails to be effective or the second eye is affected. Thereafter, molecular genetic analysis will prove LHON in these cases. Detailed anamnesis including pedigree analysis in combination with observance of the peripapillary microangiopathic alterations at the fundus will help to speed up the diagnosis of LHON, but even after exact clinical and molecular genetic diagnosis of LHON some aspects of the disease still remain a mystery today.

[Cardiorespiratory problems in the use of calcium antagonists for vasospasm prophylaxis in subarachnoid hemorrhage]. / Cardio-respiratorische Problematik beim Einsatz eines Calciumantagonisten zur Vasospasmusprophylaxe bei Subarachnoidalblutung.

Since studies by Allen et al. [1] calcium antagonists have been commonly used for prevention of cerebral vasospasm in patients suffering from acute subarachnoid hemorrhage (SAH). Vasodilatation-induced hypotension, increase of cardiac output and intrapulmonary shunting (Qs/Qt) are wellknown cardiovascular effects. These problems are discussed in light of previous reports and present case study of a 34-year-old woman treated with the calcium antagonist nimodipine after SAH. Reproducible results from invasive haemodynamic monitoring (Swan-Ganz-thermodilution catheter) indicated correlation between nimodipine application and increased intrapulmonary shunting. This effect can be hazardous for SAH patients because preexisting cerebral ischemic hypoxia makes them particularly susceptible to additional decrease in oxygen supply.

[Prognostic scores in intensive care]. / Prognostische Scores in der Intensivmedizin.

Since the development of prognostic score systems in intensive care medicine in the 1980s score models have improved substantially and are now based on much larger databases. They have been validated in many multicenter and international studies all over the world. Prognostic scoring systems may be used for assessment of severity of illness, stratifying patients prior to randomization in clinical trials, evaluation and comparing outcome and survival (hospital mortality), quality assessment, cost-benefit analysis, and in clinical decision making. Validated time points for predicting hospital mortality of ICU patients are at admission and at 24 hours. The relationship of the observed hospital mortality rate to the estimated mortality provides the basis for clinical performance measurement. Since each ICU serves a different patient population, each score system must be calibrated in the individual hospital to ensure that the model is applicable. General scores covering more than one disease are Acute Physiology And Chronic Health Evaluation (APACHE II, APACHE III), Simplified Acute Physiology Score (SAPS) and Mortality Predicting Model (MPM). The Therapeutic Intervention Scoring System (TISS) and in part the Hannover Intensive Score (HIS) evaluate exclusively the amount of medical therapy required. The TISS-Score might serve as a possible measure of resource use for the ICU portion of the hospital stay. Disease (e.g. Trauma Score, Injury of Severity Score) and patient (e.g. PRISM = Pediatric Risk of Mortality) specific scores take into account the influence of disease and patient population in relation to outcome. They are not always of more predictive value than general score models. Score models have been criticized for a number of reasons. Outcome of ICU therapy should incorporate not only survival but should also take into account quality of life, morbidity and disability. Severity scores have no role in clinical decision making for an individual patient (e.g. patient triage for ICU admission, discharge criteria, withdrawal of life support). This is due to the current low sensitivity. Subsequent validation of variables could improve the sensitivity and the value of severity scoring in the future. Nevertheless, illness severity scores will never be indicative of absolute irreversibility of disease or impossibility of survival. Advances in computer technology should assist in achieving many of the future goals of prognostic scoring systems. Most of the physiological data are available from ICU monitors and computerized laboratory systems. By electronically interfacing with the ICU monitor an automated patient data entry is possible and will provide that prognostic scores can be made available to the clinician daily.

[Emergency treatment in clinical practice in incidents following injections]. / Notfallbehandlung in der Praxis bei Zwischenfällen nach Injektionen.

A treatment concept is presented which is applicable in particular to emergencies in the office following parenteral drug administration. For physicians with office practices such events are particularly serious, for one thing because they are left to their own devices and may no longer be familiar with emergency procedure, and for another because if insurmountable complications occur the patient's and his/her family's suspicions of incorrect treatment--irrespective of possible litigation--often cannot be convincingly refuted even when they are unfounded. The paper describes the authors' own, jointly revised emergency concept for an orthopedic practice with a broad treatment spectrum. It will probably be generally applicable wherever undesirable side effects following injections--in particular after nerve block--are likely to occur.