Selective sequestration of iridoid glycosides from their host plants in Longitarsus flea beetles.

We investigated in eight species of the flea beetles genus Longitarsus (Coleoptera, Chrysomelidae) whether the beetles take up iridoid glycosides from their host plants of the Lamiaceae, Plantaginaceae, and Scrophulariaceae. Five of the beetle species, L. australis, L. lewisii, L. melanocephalus, L. nigrofasciatus, and L. tabidus, could be shown to sequester iridoid glycosides in concentrations between 0.40 and 1.55% of their dry weight. Eight different iridoid glycosides, acetylharpagide, ajugol, aucubin, catalpol, 8-epi-loganic acid, gardoside, geniposidic acid, and harpagide could be identified in the host plants, yet only aucubin and catalpol are sequestered by the beetles. No iridoid glycosides could be detected in the beetles if neither aucubin nor catalpol were present in the host plant, as in L. minusculus on Stachys recta (acetylharpagide only) and in L. salviae on Salvia pratensis (no iridoid glycosides). In one beetle species, L. luridus, we could not detect any iridoid glycosides although its field host, Plantago lanceolata, had considerable amounts of aucubin and catalpol plus two further iridoids. The five sequestering Longitarsus species differ in their capacity to store the compounds and in their affinity for catalpol relative to aucubin.

Continuous biogas production from fodder beet silage as sole substrate.

Since April 2000 a two-step anaerobic plant with two subsequent 500 m3 reactors has been producing biogas from fodder beet silage (pH 3.4-4.1) as the sole substrate. The plant is located at Kirchlengern near Bielefeld, Germany. Initially the reactors were inoculated with swine manure at 37 degrees C. After a start-up phase the process was sustained at pH 7.5-8.0 by feeding the silage as sole substrate with an HRT of about 55 d twice a day. Parallel to the biogas plant at Kirchlengern four one-step laboratory reactors were continuously driven at temperatures of 37 degrees C, 45 degrees C, 60 degrees C and 65 degrees C. They were fed with the same silage, but only once per day (one impulse). The organic loading rate (OLR) was adjusted to 3.9 g volatile solids (VS)/(l*d) with a concomitant hydraulic retention time (HRT) of 27 d. There was no problem with starting the reactors, but after 86 days the volumetric gas production of the 65 degrees C reactor ceased and a high amount of approximately 65 mM propionate could be determined. By decreasing the temperature down to 60 degrees C a stable reactor performance was recovered for a period of at least 600 further days. Interestingly microscopic analyses revealed that the morphology of methanogenic bacteria in the 60 degrees C was quite different from the 37 and 45 degrees C reactor exhibiting only rodlike methanogens whereas at 37 degrees C coccoid morphotypes besides rodlike methanogens were dominant. In a 55 degrees C reactor (separate experiment) a mixture of coccoid and rodlike methanogens established. During impulse feeding with 3.9 g (VS)/(l*d) it was observed that the quickest recovery of gas production, the pH, CH4 and CO2 content as well as the redox value could be observed at 37 degrees C or at 45 degrees C. Recovery of 75% gas volume (related to the value before or after impulse feeding) was obtained after 5.5 and 7.5 h of feeding time point whereas the 60 degrees C reactor needed 16 h. Slight significant differences were seen in the spectrum of volatile fatty acids (VFA) reaching at 37 degrees or 45 degrees C its maximum with 10-30 mM total VFA at 2-3 h after feeding. After this the VFA level declined to nearly zero (except for the 60 degrees C reactor). Therefore the 37 degrees C reactor was favoured. A double experiment with a second 37 degrees C reactor was started by a somewhat different inoculation procedure from the remaining 3 reactors, but revealed similar results. An impulse feeding experiment with a very high OLR of 16.5 g (VS)/(l*d) lasting 1 week offered a stable reactor performance with a peak GPR of up to 24 l/(l reactor *d) and an HRT of 5.45d. Therefore a long term operation with an HRT of only 7.5 days and an OLR of 12 g (VS)/(l*d) should be possible. By increasing the temperature no significantly different specific gas production rates and methane yields could be observed, e.g. it gave 600-7001 biogas from 1 kg VS. The corresponding methane content ranged between 62-64%. With a methane content of 63 +/- 1% a yield of 40.1 +/- 2 m3 methane/ton fresh fodder beet silage was obtained.

[The evaluation of different techniques used in the treatment of cholesteatomas (author's transl)]. / Evaluation des différentes techniques utilisées dans le traitement des cholestéatomes.

The authors study the results of surgical treatment of cholesteatomas on the basis of 748 records of chronic otitis - active or dry. Firstly at the time of eradication of lesions with a curative intention, amongst 152 cholesteatomas discovered, they noted that the result was successful in almost all cases following complete removal, contrasting with the frequent recurrence (25 to 30%) seen after other techniques. Subsequently, following closure of the ear by tympanoplasty or homograft, there was recurrence of cholesteatoma in one case out of 3 despite severe patient selection. In conclusion, the authors feel that the greatest caution should be exercised in treatment with conservative or reconstructive aims, the indications of which must be based not only on otological factors but also personal factors in the patient.

[Incus ossiculoplasty. 5 years' results]. / Ossiculoplastie d'enclume. Résultats à 5 ans.

A 5-year follow up study of functional results of transposition of the anvil was conducted in 114 patients. Audiometric gains obtained could be rated as satisfactory since closure of more than half of the Rinne was obtained in 70% of cases and the gain was maintained over long periods.

[Bilateral parotid gland tumors. Report on 3 cases (author's transl)]. / Tumeurs bilatérales de la glande parotide. A propos de 3 observations.

Three cases of rarely observed bilateral parotid gland tumors are reported. In two cases the tumors were of different histological types (mixed tumor on one side and an adenocarcinoma on the other side) while in the third case they were both of the mixed variety. No clear explanation for the possible bilateral nature of these tumors was found, diagnosis being made either during the initial examination or after an interval of several years.

[Comparison of a series of tympanic homografts alone and moulded autografts]. / Comparaison d'une série d'homogreffes de tympan seul et d'autogreffes moulées.

Anatomical and functional results were compared in a series of 60 tympanic homografts alone with those obtained after 80 moulded autografts. Anatomically, the moulded autografts were definitely superior complications of microperforations in approximately one-third of cases. However, from the functional point of veiw, homografts appear to provide better results, particular when the ossicles are intact.

[Benign paroxysmal positional vertigo. Diagnosis, course, physiopathology and treatment]. / Vertige paroxystique positionnel bénin. Diagnostic, évolution, physiopathologie et traitement.

The benign paroxystic positional vertigo (BPPV) is defined by brief attacks of rotational vertigo and concomitant rotatory-linear nystagmus, triggered by rapid extension and by lateral head tilt toward the affected ear. Cupulolithiasis of the posterior semi-circular canal from one inner ear is responsible of this peculiar type of positional vertigo. Definition diagnostic criteria are based upon the time history of the burst of rotational vertigo associated with the typical positioning nystagmus induced by rapid position changes from the setting to the head hanging right or left position during the Nylen-Barany maneuver. The nystagmus, which is a torsional nystagmus of the eye-balls rolling on the orbit's ground toward the affected and undermost ear, begins a few seconds after the head tilt, is best seen with Frentzel's glasses, preventing the suppression of the nystagmus by fixation, and the repetition of the maneuver will result in everlasting symptoms. According to the largest series of the literature the BPPV is the most frequent recognized cause of vertigo, with more than 30% of the cases of vertigo, explored in a dizziness clinic. In half of the cases, a peculiar etiology can be found, such as head trauma, ototoxic drug, or viral labyrinthitis, with a sex-ratio of one. On the contrary the sex-ratio among the idiopathic forms of BPPV is of two females for one male. The prognosis of BPPV is benign because of its recovery within weeks or months, either spontaneously, or after the liberatory maneuver. In rare cases, the vertigo recurs or persists for months or years. For some very handicapping BPPV, a surgical treatment can be proposed.

Production of cardenolides versus sequestration of pyrrolizidine alkaloids in larvae ofOreina species (Coleoptera, Chrysomelidae).

Adult leaf beetles of the genusOreina are known to be defended either by autogenously produced cardenolides or by pyrrolizidine alkaloids (PAs) sequestered from the food plant, or both. In this paper we analyze larvae of differentOreina species and show that the larvae contain the same defensive toxins as the adults in quantities similar to those released in the adults' secretion. Both classes of toxins are found in the body and hemolymph of the larvae, despite their different origins and later distribution in the adults. Larvae of sequestering species differed in their PA patterns, even though they fed on the same food plants. The concentration in first-instar larvae of a PA-sequestering species was similar to that in fourth-instar larvae. In all stages examined, the amount of PAs per larva did not greatly exceed the estimated uptake of one day. Eggs of two oviparous species contained large concentrations of the adult's toxins, while neonates of a sequestering larviparous species had no PAs.

Resolving phylogeny at the family level by mitochondrial cytochrome oxidase sequences: phylogeny of carrion beetles (Coleoptera, Silphidae).

We investigated the phylogenetic relationships of carrion beetles (Coleoptera, Silphidae) using 2094 bp of their mitochondrial cytochrome oxidase subunit I and II and tRNA leucine gene sequences. Shorter fragments of this gene region previously have been used to establish generic relationships in insects. In this study, they provided more than sufficient resolution, although the third positions of the protein-coding sequences reached saturation for the deeper divergences. This first published phylogeny for the Silphidae comprises 23 species from 13 genera sampled across the geographic range of the family. In addition, we included species from three related families as outgroups. One of these families, the Agyrtidae, was, until recently, included in the Silphidae, but its resolution here justifies its current position as a separate family. The silphid subfamilies Nicrophorinae and Silphinae are monophyletic in all analyses. All genera for which several species were sampled are supported as monophyletic groups, with the exception of the genus Silpha. European and North American representatives of two Nicrophorus species described from both continents are supported as each others' closest relatives. The lineage that colonized Gondwanaland and that most likely originated in the Palearctic is the most basal within the Silphinae.

Host use evolution in Chrysochus milkweed beetles: evidence from behaviour, population genetics and phylogeny.

In two sister species of leaf beetles with overlapping host associations, Chrysochus auratus and C. cobaltinus, we established diet breadth and food preference of local populations for evaluation together with genetic differentiation between populations. While C. auratus turned out to be monophagous on the same plant wherever we collected the beetles, the studied populations of C. cobaltinus fed on three different plant species in the field. Plant preference and ranking of the potential host plants significantly differed between these populations. The amount of genetic differentiation between populations was measured by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay of a 1300 bp mitochondrial DNA (mtDNA) sequence. In addition, the dominant genotypes of all populations were sequenced. No genetic differentiation between the populations of C. auratus could be detected in the RFLP assay and sequence divergence was low (= 0.3%). In C. cobaltinus, on the other hand, genetic differentiation between populations was high, revealing a lack of gene flow over a much smaller scale and a maximum of 1.3% sequence divergence. C. cobaltinus thereby has the prerequisites for host race formation on different plants from the original host spectrum. Our sequence-based phylogeny estimate allows us to reconstruct historical diet evolution in Chrysochus. Starting from an original association with Asclepiadaceae, the common ancestor of C. auratus and C. cobaltinus included Apocynaceae in its diet. The strict specialization on Apocynum and the loss of acceptance of Asclepiadaceae observed in C. auratus could have resulted from a process similar to that displayed by C. cobaltinus populations.

Phylogeny of the genus Aleochara inferred from mitochondrial cytochrome oxidase sequences (Coleoptera: Staphylinidae).

The phylogeny of the genus Aleochara was previously poorly understood due to difficulties with phylogenetic reconstruction by morphological characters. We present here a phylogeny based on the sequences of a 2022-bp fragment of the COI/II genes; 50 Aleochara and 10 outgroup species were included in the analysis. We used parsimony, minimum-evolution, and maximum-likelihood analyses to infer the phylogeny of the group. Our data do not support the commonly assumed sister group relationship between Aleocharini and Hoplandriini. Aleochara is resolved as a monophylum, although A. clavicornis might not belong to the genus. Within Aleochara, there are two large monophyletic clades. Many of the existing subgenera are shown to be para- or polyphyletic; others are likely to be monophyletic. Tinotus morion, previously assigned to the Hoplandriini, is strongly supported as belonging to Aleochara. According to our data, the mesosternal carina that has been used as an important character for classification has arisen and been reduced independently in several clades within Aleochara.

Uptake and metabolism of pyrrolizidine alkaloids in Longitarsus flea beetles (Coleoptera: Chrysomelidae) adapted and non-adapted to alkaloid-containing host plants.

Several Longitarsus flea beetle species sequester pyrrolizidine alkaloids acquired from their Asteraceae and Boraginaceae host plants. We carried out feeding and injection experiments using radioactively labeled pyrrolizidine alkaloids to investigate the physiological mechanisms of uptake, metabolism and storage of alkaloids in adult beetles. We examined six Longitarsus species belonging to different phylogenetic clades in a comparative approach. All species that accepted pyrrolizidine alkaloids in a preceding food choice study showed the ability both to store pyrrolizidine alkaloid N-oxides and to metabolize tertiary pyrrolizidine alkaloids into their N-oxides. Regardless of whether the beetles' natural host plants contain pyrrolizidine alkaloids or not, these species were found to possess an oxidizing enzyme. This oxygenase appears to be specific to pyrrolizidine alkaloids: [3H]Atropine and [14C]nicotine, two alkaloids not related to pyrrolizidine alkaloids, were neither stored nor N-oxidized by any of the tested species. One species, L. australis, that strictly avoids pyrrolizidine alkaloids behaviorally, exhibited a lack of adaptations to pyrrolizidine alkaloids on a physiological level as well. After injection of tertiary [14C]senecionine, beetles of this species neither N-oxidized nor stored the compounds, in contrast to L. jacobaeae, an adapted species that underwent the same treatment. L. jacobaeae demonstrated the same efficiency in N-oxidation and storage when fed or injected with tertiary [14C]senecionine.

Pyrrolizidine alkaloids in Pulmonaria obscura.

Leaves, roots and rhizomes, and inflorescenses of Pulmonaria obscura from two localities and of two different growth forms within one site were analyzed by GC-MS for their content in pyrrolizidine alkaloids (PAs). In roots and rhizomes PAs of the lycopsamine type typical for the Boraginaceae could be detected, e. g. intermedine, lycopsamine, and their O(7)-derivatives. The total PA concentrations in roots and rhizomes lay between 0.026 and 0.158 mg/g dry weight. In leaves and inflorescenses, on the other hand, only trace amounts of PAs (below 0.4 ng/mg dry weight) could be detected. No significant differences in total concentration of PAs could be found between the two sites.

Comparison between caloric and canal impulse rotatory test.

It has been suggested that the rotatory test using videosnystagmoscopy can also be applied for identification of vestibular canal paresis. In this study, we test this hypothesis by comparing the results of a canal impulse rotatory test with those of the caloric test using the method described by Freyss. Our study indicates that only in 38% of all tested patients, the same results between the two tests have been obtained. Furthermore, which the group with abnormal findings is observed separately, in 24% patients only these two test methods lead to the same results. Consequently, the canal impulse rotatory test cannot replace the caloric test.

Distribution of autogenous and host-derived chemical defenses inOreina leaf beetles (Coleoptera: Chrysomelidae).

The pronotal and elytral defensive secretions of 10Oreina species were analyzed. Species feeding on Apiaceae, i.e.,O. frigida andO. viridis, or on Cardueae (Asteraceae), i.e.,O. bidentata, O. coerulea, andO. virgulata, produce species-specific complex mixtures of autogenous cardenolides.O. melanocephala, which feeds onDoronicum clusii (Senecioneae, Asteraceae), devoid of pyrrolizidine alkaloids (PAs) in its leaves, secretes, at best, traces of cardenolides. Sequestration of host-plant PAs was observed in all the other species when feeding on Senecioneae containing these alkaloids in their leaves.O. cacaliae is the only species that secretes host-derived PA N-oxides and no autogenous cardenolides. Differences were observed in the secretions of specimens collected in various localities, because of local differences in the vegetation. The other species, such asO. elongata, O. intricata, andO. speciosissima, have a mixed defensive strategy and are able both to synthesize de novo cardenolides and to sequester plant PA N-oxides. This allows a great flexibility in defense, especially inO. elongata andO. speciosissima, which feed on both PA and non-PA plants. Populations of these species were found exclusively producing cardenolides, or exclusively sequestering PA N-oxides, or still doing both, depending on the local availability of food-plants. Differences were observed between species in their ability to sequester different plant PA N-oxides and to transform them. Therefore sympatric species demonstrate differences in the composition of their host-derived secretions, also resulting from differences in host-plant preference. Finally, within-population individual differences were observed because of local plant heterogeneity in PAs. To some extent these intrapopulation variations in chemical defense are tempered by mixing diet and by the long-term storage of PA N-oxides in the insect body that are used to refill the defensive glands.

Comparison between caloric and canal impulse rotatory test.

It has been suggested that the rotatory test using videonystagmoscopy can also be applied for identification of vestibular canal paresis. In this study, we test this hypothesis by comparing the results of a canal impulse rotatory test with those of the caloric test using the method described by Freyss. Our study indicates that only in 38% of all tested patients, the same results between the two tests have been obtained. Furthermore, when the group with abnormal findings is observed separately, in 24% patients only these two test methods lead to the same results. Consequently, the canal impulse rotatory test cannot replace the caloric test.

Study of the efficacy and safety of ciprofloxacin in the treatment of chronic sinusitis.

Over a 12-month period, 56 patients with acute exacerbations of chronic sinusitis were included in this study. The mean age of the patients was 48.6 +/- 10.1 years. The period between the onset of sinus symptoms and inclusion in the study was 10.1 +/- 12.1 years and the exacerbation had lasted 2.4 +/- 4.4 months. Patients were given 500 mg ciprofloxacin twice daily for 9 days. Bacteriological specimens taken at inclusion were positive in 53.6% of cases, 25 Gram-positive and 16 Gram-negative pathogens being isolated. At the end of the treatment, rhinorrhoea, which was the main criterion for evaluation of the efficacy of treatment, had disappeared in 46/54 (85%) patients who were examined. Of the 30 patients whose sinusitis was confirmed by bacteriological test, the bacteria were eradicated in 90% of cases. At the end of a 4-week follow-up period after the end of treatment, the clinical success rate was 74.5%, the partial success rate 21.2% and the failure rate 4.3%. There were no bacteriological recurrences. Ten adverse events were observed in seven patients and in four (7%) of these treatment had to be withdrawn. This study showed ciprofloxacin to be an effective and well tolerated treatment for exacerbations of chronic sinusitis treated in non-hospital practice.

Study of the efficacy and safety of ciprofloxacin in the treatment of chronic otitis.

Over a period of 18 months 76 patients presenting with acute exacerbations of chronic otitis were enrolled in this study. The mean age of the patients was 46.4 +/- 16.8 years, the interval between the onset of symptoms of otitis and enrollment in the study was 22.1 +/- 19.4 years and the infective episode had been developing for 36.4 +/- 72.4 days. The patients received either 500 or 750 mg ciprofloxacin twice daily for a period of 9 days with no topical antibiotic treatment. The bacteriological specimen taken at enrollment was positive in 93% of the patients and led to the isolation of 85 microorganisms, 65% of which were Pseudomonas aeruginosa, Staphylococcus aureus or Proteus mirabilis. At the end of the treatment period, discharge from the ears (the primary clinical evaluation criterion) had ceased in 44/69 (64%) patients examined. The clinical cure rate was 23/39 (59%) for patients treated with 500 mg ciprofloxacin twice daily and 21/30 (70%) for those treated with 750 mg twice daily (difference was not statistically significant). At the end of a 4-week post-treatment follow-up, the clinical relapse rate was 7%. There was no relapse in the patients with chronic otitis which had been bacteriologically documented. Eight adverse effects were observed in five patients and treatment had to be discontinued in one case. This study shows ciprofloxacin to be an effective and safe agent for use in the treatment of acute exacerbations of chronic otitis treated in general practice. In certain cases, however, additional treatment with a topical antibiotic may be desirable.