Effect of the digenean parasites of fish on the fauna of Mediterranean lagoons.

Attention is drawn to the effects of parasites on their hosts, taking as a model the digenean parasites of teleosts (hereafter: fish) from lagoons along the French Mediterranean coast. Because digeneans have a heteroxenic life cycle, their impact is not limited to the definitive host, which harbours the sexual adults, but is extended to the first host (mollusc) and to the second host ("invertebrate" or fish). Adult parasites, in order to ensure efficient sexual reproduction, never cause excessive damage to their definitive host, usually only exploiting the intestinal fluids; however, the host must intensify its search for prey, which results in a diminished fitness. Within the first host, 'larval' stages of digenean parasites invade the gonads, resulting in its castration, then exhaustion and eventually death. The diversion of energy from the second hosts towards the parasites forces them to intensify their search for food, resulting in decreased fitness and an increased risk of being eaten; in addition, manipulation of the host's behaviour by parasites drives this host into the food chain of the definitive host. In lagoons, many individuals of almost all species of fish and invertebrates act as first, second and/or definitive hosts for digeneans. Obviously, parasites have a severe impact on the population dynamics of key taxa, on the food web and therefore also on the functioning of the whole lagoon ecosystem. Yet this impact has been largely overlooked or underestimated in functioning models, by ecologists, who tend to prioritize more apparent trophic relationships.

The ZNF35 human zinc finger gene encodes a sequence-specific DNA-binding protein.

We developed a rapid method to determine DNA-binding sites for putative DNA-binding proteins. This procedure has been successfully used to define a specific consensus site for the human ZNF35 zinc finger gene. ZNF35 encodes a 58-kDA polypeptide containing 11 consecutive finger motifs located at the amino terminus, and an acidic domain located at the carboxy terminus. These features suggest that ZNF35 is a site-specific DNA-binding protein involved in the regulation of gene expression. We have expressed the ZNF35 protein from E. coli and have employed a Southwestern-polymerase chain reaction method using random oligonucleotides to identify its high-affinity binding site. The core sequence for the ZNF35 protein-binding site is 5'-C/GC/GAAG/TA-3'.

Methodology of head-up tilt testing potentiated with sublingual nitroglycerin in unexplained syncope.

Shortened head-up tilt testing (HUT) potentiated with sublingual nitroglycerin (60 degrees passive standing for 20 minutes followed, if negative, by 400 microg of sublingual nitroglycerin spray with the test continuing for another 20 minutes) differs from conventional nitroglycerin HUT for a shorter drug-free phase (20 vs 45 minutes). To compare the positivity rate of the 2 protocols, both tests were performed in a randomized sequence in 10 patients with unexplained syncope (study 1), and another 42 patients were randomly assigned either to conventional or to shortened nitroglycerin HUT (study 2). To evaluate the reproducibility of the shortened nitroglycerin HUT, another 38 patients with unexplained syncope underwent 2 consecutive tests within a 7+/-8 day interval (study 3). Finally, to evaluate the specificity of the test, 47 control subjects underwent shortened nitroglycerin HUT (study 4). Seven positive responses were observed during shortened nitroglycerin HUT, and there were 8 positive responses during conventional nitroglycerin HUT (p = NS) in the study 1 group. Fifteen positive (71%) responses, 5 negative responses, and 1 exaggerated response were observed during shortened nitroglycerin HUT; 16 positive (76%, p = NS vs. shortened nitroglycerin HUT), 3 negative, and 2 exaggerated responses were observed during conventional nitroglycerin HUT in the study 2 group. During the first test, 21 patients (55%) had a positive, 15 patients had a negative, and 2 patients had an exaggerated response in study group 3. During the second test, 15 positive (39%), 19 negative, and 4 exaggerated responses were observed. Thus, the reproducibility was 67% for a positive and 94% for a negative test. In control subjects, 2 positive (4%) responses, 38 negative, and 7 exaggerated responses were observed with a specificity of 96% in study group 4. In patients with unexplained syncope, shortened nitroglycerin HUT allowed a positivity rate similar to that of the conventional test. Moreover, the shortened test provided a high specificity and adequate reproducibility for both the positive and the negative responses.

Effects of chronic vasodilator therapy to enhance susceptibility to vasovagal syncope during upright tilt testing.

To evaluate the effect of chronic vasodilator therapy on susceptibility to vasovagal syncope, 45 patients with syncope and a positive response to tilt testing were randomly assigned to continue or to discontinue vasodilators. The study result demonstrated that chronic vasodilator therapy enhances susceptibility to vasovagal reaction during upright tilt testing.

The life cycle of Opecoeloides columbellae (Pagenstecher, 1863) n. comb. (Digenea, opecoelidae): evidence from molecules and morphology.

The cosmopolitan digenean family Opecoelidae comprises several hundred species, whose adults live in the digestive tract of marine and freshwater fishes. The genus Opecoeloides Odhner, 1928 is represented in the Mediterranean by a single species, Opecoeloides furcatus (Bremser in Rudolphi, 1819), that has been recorded from six definitive hosts species. To see if this broad host range could be the result of an underestimation of species diversity, we obtained ITS1 ribosomal DNA sequences as well as morphological data from adult specimens of O. furcatus isolated from two definitive hosts species: Mullus surmuletus and Gaidropsarus mediterraneus. Sequence and morphological data were also obtained from several opecoelid cercariae and metacercariae occurring in different invertebrate hosts. The data presented here provide striking evidence that O. furcatus specimens isolated from the two host fishes represent distinct species. This argument is reinforced by the fact that cercariae corresponding to each of these adult species were found in two molluscan host-species, Columbella rustica and Mitrella scripta. These parasite species differ by several nucleotide substitutions and a 60 bp-long insertion in the ITS1. They also show clear morphological differences in testis and ovary shape, as well as in their mean dimensions. Here, we attribute the adult specimens found in G. mediterraneus to Opecoeloides columbellae (Pagenstecher, 1863) n. comb. This species was described and compared with O. furcatus from M. surmuletus. ITS1 sequence comparison allowed identification of the cercaria (occurring in C. rustica) and metacercaria (occurring in Hippolyte inermis) of O. columbellae n. comb.

Molecules, morphology and morphometrics of Cainocreadium labracis and Cainocreadium dentecis n. sp. (Digenea: Opecoelidae) parasitic in marine fishes.

Molecular, morphological and morphometric analyses were conducted on several samples of Cainocreadium labracis (Opecoelidae), a trematode parasitic in marine teleosts. The samples were isolated from several specimens of Dicentrarchus labrax, the type host, and Dentex dentex. The molecular analysis of complete Internal Transcribed Spacer sequences of ribosomal DNA revealed that specimens isolated from each host species form two well-defined groups, whose sequence divergence reaches 7.5%. The morphological study showed that the two groups can be distinguished by several characters, including the level of maximum body breadth, the relative position of the testes, the shape of the cirrus pouch, and the extent of the uterus. Multivariate analyses of morphometrics demonstrated consistency of most of the characters for discriminating the two groups. Our results show that C. labracis specimens isolated from D. labrax and D. dentex represent clearly distinct entities from molecular, morphological and statistical points of view, which has enabled us to describe a new species, Cainocreadium dentecis n. sp.

Molecular identification of developmental stages in Opecoelidae (Digenea).

Nuclear ribosomal DNA sequences represent a useful tool for distinction of poorly differentiated developmental stages, such as trematode cercariae or metacercariae. Here, the complete internal transcribed spacer region of the ribosomal DNA (ITS 1 + 5.8S + ITS 2) was sequenced for 29 specimens of the digenean family Opecoelidae, including 16 adult specimens and 13 undescribed larval stages (nine cercariae and four metacercariae) occurring in various marine host organisms. Six cercariae and three metacercariae were found to match their corresponding adult form. This work also revealed that cercariae of the same species are able to infect more than one gastropod host species, suggesting that the specificity for the first intermediate host within the Digenea may be lower than previously thought.

Use of the ITS rDNA for elucidation of some life-cycles of Mesometridae (Trematoda, Digenea).

Identification of larval stages is crucial for elucidating the life-cycles of various Digenea. However, in many digenean species, the larvae lack distinctive morphological features and it is impossible to establish the affiliation between the larval and adult stages by using morphological criteria. Molecular methods, based on DNA sequencing or PCR-restriction fragment length polymorphism analysis, can offer a new tool for larval-stage identification. In this study, the sequences of internal transcribed spacer of the ribosomal DNA were used to identify the cercariae of three out of five species of the family Mesometridae (Centroderma spinosissima, Elstia stossichianum and Wardula capitellata). The three species differ from one another by number of repeats in the region of internal transcribed spacer 1. The phylogeny of Mesometridae was inferred from their internal transcribed spacer ribosomal DNA sequences. The PCR-linked restriction fragment length polymorphism approach was developed for future life-cycle and ecological studies of this family.

Pheochromocytoma crisis caused by contemporary ergotamine, caffeine, and nimesulide administration.

Pheochromocytoma is a rare tumor that secretes excess catecholamines. Pheochromocytoma crises may be precipitated by the use of several drugs. This article describes the case of a patient affected by pheochromocytoma in whom multiple organ failure developed after contemporary administration of ergotamine, caffeine, and nimesulide. The patient recovered completely long after surgical intervention.

The life cycle of Monorchis parvus (Digenea: Monorchiidae) demonstrated by developmental and molecular data.

Cercaria cerastodermae I, a digenean parasite of Cerastoderma edule, was recorded for the first time in the Atlantic Ocean off the Iberian peninsula. Sporocysts were present in the hemolymph of the digestive gland, gonad, gills, and foot of the mollusc. Most of the cercariae present within sporocysts were encysted as metacercariae. The corresponding adult stages were obtained after experimental infection of several Diplodus sargus artificially reared in fish farms and that had previously been protected against natural infections. Numerous adult specimens of Monorchis parvus were collected in Diplodus annularis along the French Mediterranean coast. Comparison of wild and experimental adults allowed the adult stage of Cercaria cerastodermae I to be identified as M. parvus. Another monorchid, Monorchis monorchis, a parasite of Spondyliosoma cantharus, was found in the same Mediterranean area and compared with M. parvus. Additionally, ITS1 nuclear ribosomal DNA sequences of C. cerastodermae I and of the adults collected in naturally infected D. annularis and S. cantharus were obtained. Sequence data indicate that C. cerastodermae I corresponds to the adult of M. parvus found in D. annularis and is clearly distinct from M. monorchis found in S. cantharus.

Assessment of captopril and nicardipine effects on chronic occlusive arterial disease of the lower extremity using Doppler ultrasound.

The effects of two potent vasodilating drugs, captopril (C) (25 mg tid), nicardipine (N) (20 mg tid), and placebo (P) were evaluated and compared, in 10 men (mean age of sixty-five years) with intermittent claudication from moderate to severe multilevel chronic occlusive arterial disease (COAD) of the lower extremity, by use of the Doppler ultrasonic method, at rest and after Carter's exercise test. All the examined subjects were normotensive, without diabetes or cardiopathy; all have been smokers. The eight-week total protocol consisted of an initial two-week placebo run-in period followed by two active drug phases and a two-week placebo phase, according to a double-blind, randomized, crossover design. At the end of each two-week period, ankle-arm index (AAI) and, following exercise, onset of lower extremity discomfort time (ODT), duration of exercise (ET), decrease of ankle systolic pressure after test (APD), and recovery time (RT) were determined. Moreover, at rest, just after exercise, and after recovery, simultaneous common femoral artery velocity waves were recorded and analyzed by a quantitative approach to detect the peripheral vasomotor adjustments. None of the patients required the withdrawal of the active treatments. Compared with P, C significantly reduced APD and RT, and N reduced RT and AAI; furthermore N caused a significant decrease in ODT, whereas C showed a trend, although not statistically significant, to increase ODT. Neither active therapy modified ET. These results suggest that C and N have different short-term effects on peripheral circulation in COAD. During exercise, C induces hemodynamic improvement in the ischemic lower extremity probably by inhibition of the sympathetic system and consequent reduction in collateral vessel vasoconstriction.

Use of Doppler flow velocity waveform analysis in detection of initial diabetic microangiopathy.

Doppler flow velocity waveform analysis (FVWFA), recorded from the dorsalis pedis artery (DPA) and the radial artery (RA), was performed on 36 women in attempting to detect an initial diabetic microangiopathy (DM). The study comprised two groups of women affected by non-insulin-dependent diabetes mellitus, 6 patients (pts) of reproductive age (1), 12 pts in menopause (II), and two groups of age-matched healthy controls (C) (III and IV). Clinical signs of initial DM were present in group I. All the examined pts were nonsmokers and normotensive and without cardiopathy, signs of diabetic macroangiopathy, collagen vascular disease and/or Raynaud's phenomenon, and renal failure. Four waveform dimensions capable of separating different degrees of peripheral obstructive arteriolar disease were determined on velocity tracing and the results used in a single best discriminant equation. The resultant discriminant score (DS), derived by FVWFA on DPA, showed a highly accurate rate of separating the young pts with DM from both C and the pts in menopause without DM. Furthermore, the resultant DS was statistically not different in groups II, III, and IV. In conclusion, FVWFA on DPA, in this experience, has proved to be an accurate and sensitive method in the detection of initial DM.

[Right atrial metastasis as an initial clinical manifestation of adenocarcinoma of the colon]. / Metastasi atriale destra come prima manifestazione clinica di adenocarcinoma del colon.

We report the case of a young woman suffering from colonic mucinous adenocarcinoma with the unusual clinical presentation as a right atrial metastasis. The tumor was successfully resected, and after histopathological characterization, the primary lesion was searched for and discovered. The tumor had the clinical appearance and echocardiographic pattern of a pedunculated atrial myxoma. The aspects of echo-patterns useful for differential diagnosis are discussed.

Molecular phylogeny of Mesometridae (Trematoda, Digenea) with its relation to morphological changes in parasites.

Complete ITS (Internal Transcribed Spacer) ribosomal DNA sequences were obtained for the six species know at present time within the Mesometridae Poche, 1926. The adult stages are intestinal parasites of herbivorous sparid teleosts. Aligned sequences were analysed with Maximum Parsimony, Maximum Likelihood and Neighbor-Joining phylogenetic methods to infer evolutionary relationships among mesometrid species. The ITS-based phylogeny obtained showed the two Wardula species as a sister group to other Mesometridae, and as compared to morphological data, suggest some general tendencies in the morphological evolution of this group. It consists mainly in changes from elongated to subcircular forms, regression of the pharynx, and the development of a strong accessory attachment organ.

[Original adaptive characters of intestinal Digenea of Sarpa salpa (Teleostei, Sparidae) and their interpretation in terms of evolution]. / Caractères adaptatifs originaux des Digènes intestinaux de Sarpa salpa (Teleostei, Sparidae) et leur interprétation en termes d'évolution.

In the family Sparidae, the genus Sarpa is distinguished by a few characteristics: monospecificity, vegetarian diet and wide geographical distribution. The helminth fauna of Sarpa salpa is also very original. Indeed, the digenean parasites of this Teleostean fish are essentially classified into two families restricted to this fish. In the present paper, the author redescribes Mesometra orbicularis, M. brachycoelia, Centroderma spinosissima, Elstia stossichianum, Wardula capitellata (family Mesometridae) together with Robphildollfusium fractum (family Robphildollfusidae). Various original and yet unknown features are pointed out. Among these unusual structures, several correspond to adaptive characteristics favouring the settlement of the Digenean on the peculiar digestive gut wall of this herbivorous fish. Indeed, the intestinal mucous membrane of Sarpa salpa exhibits very few villi giving it an unusual smooth aspect. Therefore, the Mesometridae which always have just a single sucker (monostomatous) have selected a new kind of compensatory adhesive structure. Sometimes, the anterior end of the body becomes a sucker due to the particular distribution of the muscle strings; in other examples, the whole body becomes a sucker and its edges become considerably thinner to improve the tightness of the adhesive system. Other original anatomical features have been selected to allow survival in a medium rich in plant detritus. So, in the oral sucker crests ornemented by numerous sclerous denticles seem to act as a microfilter for the intestinal chyme in which plant fibres predominate. The original pharynx seems to act as a suction-force pump. The excretory system, which is of a reticular type, penetrates the whole parenchyma and this could be a response to huge intestinal fermentations. The Digenea of Sarpa salpa are not interpreted by the author as true parasites but as endocommensal symbionts. These inquiline species are not immunogenic, or at least only slightly so, since they do not feed upon the host itself but upon its intestinal chyme. In most cases this results in a high parasite density (post larvae and adults) together with a cohabitation of the various species along the various intestinal segments. Coexistence of several species, systematically very close, evidently raises the question of their reproductive isolation. The author proposes an answer founded upon data of allopatric speciation.

[Gymnophallus rebecqui n. sp. (syn. Parvatrema sp. 1, J. Rebecq, 1964) (Digenea: Gymnophallidae), an intestinal parasite of ducks from Camargue (France)]. / Gymnophallus rebecqui n. sp. (syn. Parvatrema sp. 1, J. Rebecq, 1964) (Digenea: Gymnophallidae) parasite intestinal d'anatidés de Camargue (France).

Gymnophallus rebecqui n. sp. replace Parvatrema sp. 1 J. Rebecq, 1964. Metacercariae are described. They occur free in the extrapallial space of Cerastoderma glaucum and Abra ovata, at the central part of the valves. Pallial epithelium hypertrophy is induced by metacercariae and damages are produced at the inner face of the shell of Abra ovata. Adults have been reared in laboratory hosts (Aythya ferina, A. fuligula, Anas platyrhynchos, Tadorna tadorna and Larus argentatus michaellis). Natural adults have been discovered in the anterior and median gut of Aythya ferina, A. fuligula and Anas clypeata. Adults are described and compared with other related Gymnophallid species. This new species is not a member of Parvatrema but belongs to Gymnophallus genera.

[The larval stages of Meiogymnophallus strigatus (M. V. Lebour, 1908) n. comb. (Trematoda: Gymnophallidae)]. / Les stades larvaires de Meiogymnophallus strigatus (M. V. Lebour, 1908) n. comb. (Trematoda: Gymnophallidae).

Cercariae and metacercariae of Meiogymnophallus strigatus (M. V. Lebour, 1908) have been found in some places along the coast of Camargue (South of France). Sporocysts and cercariae are parasites of Tellina tenuis. Metacercariae occur in various Bivalves, mainly in Donax trunculus and D. semistriatus. Cercariae and metacercariae are described. Relationship between cercariae and metacercariae is demonstrated. Metacercarial microhabitat is delimited. Reaction of D. trunculus in front of the metacercarial infection is studied. Some metacercariae are free-living in the extrapallial space, between the mantle and the shell. These larvae have an advanced genital maturity. Most of the metacercariae are massed together and are enclosed by the host's epithelium. Their excretory vesicle, which is packed with blackish granules, almost fills the entire body. Attempts to infect laboratory reared birds by feeding metacercariae from Donax trunculus were not successful. The natural final host is unknown. In consideration of the metacercarial anatomy, Cercaria strigata M. V. Lebour, 1908 is placed in the genus Meiogymnophallus H. L. Ching, 1965.