Exposure to copper induces oxidative and stress responses and DNA damage in the coral Montastraea franksi.

Copper is a common chemical contaminant in coastal environments, including coral reefs. Ecotoxicological studies have demonstrated that exposure to copper can cause stress and detrimental effects in both host cnidarian and algal symbionts. The objective of this study was to investigate the sublethal effects of copper on the reef-building coral Montastraea franksi, by identifying genes with altered expression in corals exposed to dissolved copper, and by measuring the extent of damage to DNA in response to copper exposure. Corals exposed to 30 µg L(-1) copper for 48 h experienced significant DNA damage and displayed changes in expression patterns of genes that are known to play role cellular and oxidative stress responses. Corals also experienced changes in gene expression of genes that are not already known to play roles in oxidative stress in corals. Our data suggest that these genes may either play roles directly in mediating a stress response, or may be genes acting downstream of the stress response. These include an ETS domain-containing transcription factor related to the ETS1 family of transcription factors, known in mammals to mediate development, disease, and stress response, and two genes that are associated with biomineralization: galaxin, a protein from the organic matrix of the coral skeleton, and a coral-specific gene SCRIP2.

Differential gene expression during thermal stress and bleaching in the Caribbean coral Montastraea faveolata.

The declining health of coral reefs worldwide is likely to intensify in response to continued anthropogenic disturbance from coastal development, pollution, and climate change. In response to these stresses, reef-building corals may exhibit bleaching, which marks the breakdown in symbiosis between coral and zooxanthellae. Mass coral bleaching due to elevated water temperature can devastate coral reefs on a large geographical scale. In order to understand the molecular and cellular basis of bleaching in corals, we have measured gene expression changes associated with thermal stress and bleaching using a complementary DNA microarray containing 1310 genes of the Caribbean coral Montastraea faveolata. In a first experiment, we identified differentially expressed genes by comparing experimentally bleached M. faveolata fragments to control non-heat-stressed fragments. In a second experiment, we identified differentially expressed genes during a time course experiment with four time points across 9 days. Results suggest that thermal stress and bleaching in M. faveolata affect the following processes: oxidative stress, Ca(2+) homeostasis, cytoskeletal organization, cell death, calcification, metabolism, protein synthesis, heat shock protein activity, and transposon activity. These results represent the first medium-scale transcriptomic study focused on revealing the cellular foundation of thermal stress-induced coral bleaching. We postulate that oxidative stress in thermal-stressed corals causes a disruption of Ca(2+) homeostasis, which in turn leads to cytoskeletal and cell adhesion changes, decreased calcification, and the initiation of cell death via apoptosis and necrosis.

[Regulatory aspects of clinical trials for medical devices and in vitro diagnostics]. / Regulatorische Anforderungen an klinische Prüfungen mit Medizinprodukten und Leistungsbewertungsprüfungen mit In-vitro-Diagnostika.

The European Medical Device Directives and In Vitro Diagnostic Directive were transposed into the German Medical Device Law (MPG) in 2002. Clinical trials in Germany are to be conducted in accordance with the MPG, Harmonized European Standards, professional codes and laws and for medical and scientific reasons according to ICH GCP.

[Ethical and regulatory requirements for conducting clinical trials: patient information and patient informed consent]. / Patienteninformation und Einwilligung bei Geschäftsfähigen, Geschäftsunfähigen und Minderjährigen vor der Teilnahme an klinischen Prüfungen.

Written informed consent must be obtained from legally competent subjects, after having been duly informed verbally and in writing, before participation in a clinical trial with pharmaceutical products or medical devices. For persons or minors incapable of giving their informed legal consent, the written consent of the subject's legal representative is mandatory. The subject's presumed or expressed wish must be considered.

Localization of a symbiosis-related protein, Sym32, in the Anthopleura elegantissima-Symbiodinium muscatinei Association.

Cnidarian-dinoflagellate symbioses are widespread in the marine environment. Growing concern over the health of coral reef ecosystems has revealed a fundamental lack of knowledge of how cnidarian-algal associations are regulated at the cellular and molecular level. We are interested in identifying genes that mediate interactions between the partners, and we are using the temperate sea anemone Anthopleura elegantissima as a model. We previously described a host gene, sym32, encoding a fasciclin domain protein, that is differentially expressed in symbiotic and aposymbiotic A. elegantissima. Here, we describe the subcellular localization of the sym32 protein. In aposymbiotic (symbiont-free) hosts, sym32 was located in vesicles that occur along the apical edges of gastrodermal cells. In symbiotic hosts, such vesicles were absent, but sym32 was present within the symbiosome membranes. Sym32 (or a cross-reactive protein) was also present in the accumulation bodies of the symbionts. Although the anti-sym32 antiserum was not sufficiently specific to detect the target protein in cultured Symbiodinium bermudense cells, Western blots of proteins from two Symbiodinium species revealed a protein doublet of 45 and 48 kDa, suggesting that the symbionts may also produce a fasciclin domain protein. We suggest that host sym32 is relocated from gastrodermal vesicles to the symbiosome membrane when symbionts are taken into host cells by phagocytosis.

Acetylsalicylic acid effervescent 1000 mg (Aspirin) in acute migraine attacks; a multicentre, randomized, double-blind, single-dose, placebo-controlled parallel group study.

In this multicentre, randomized, double-blind, single-dose study a total of 374 patients generally suffering from migraine attacks suitable for treatment with non-prescription drugs, received either oral acetylsalicylic acid effervescent 1000 mg (ASAE) or effervescent placebo for the treatment of an acute migraine attack. Of the 343 patients fulfilling the criteria for efficacy analysis 169 patients took acetylsalicylic acid and 174 placebo. Response rates (reduction of headache severity from severe or moderate to mild or no pain at 2 h after administration) were 55.0% for acetylsalicylic acid and 36.8% for placebo (P < 0.001). Twenty-nine percent of patients in the active treatment group were pain-free after 2 h compared with 16.7% in the placebo group (P = 0.007). No headache recurred within 24 h post-dose in 84.6% of patients in the active group and in 85.1% of patients in the placebo group. Effervescent placebo reduced nausea and vomiting to the same degree as the active drug. Adverse events of acetylsalicylic acid (8.3%) were generally mild or moderate and comparable to those of placebo (2.9%). This study shows that oral ASAE is safe and effective for the treatment of acute migraine attacks.

Symbiosis-enhanced gene expression in cnidarian-algal associations: cloning and characterization of a cDNA, sym32, encoding a possible cell adhesion protein.

Mutualistic endosymbioses between two partners are complex associations that are regulated by the genetic interactions of the partners. One important marine symbiosis is that between various cnidarians, such as corals and anemones, and their photosynthetic algal symbionts. We have been interested in characterizing cnidarian host genes that are expressed as a function of the symbiotic state, using the temperate sea anemone Anthopleura elegantissima as a model. In this study, we report on symbiosis-enhanced expression and synthesis of sym32 in anemones. We characterized the full-length sym32 cDNA, obtained by RT-PCR, and demonstrated, by semi-quantitative RT-PCR, that sym32 transcript was much more abundant in symbiotic than in non-symbiotic host anemone RNA. Further, using immunoblots, we determined that an antibody made to a sym32 fusion protein labeled a 32 kD band much more strongly in symbiotic compared to non-symbiotic anemone protein homogenates. Databank searches revealed that the sym32 deduced amino acid sequence shares significant homology with the fasciclin I (Fas I) family of homophilic cell adhesion proteins, present in a variety of organisms ranging from bacteria to humans. This strong homology with the Fas I family suggests that sym32 is involved in regulation of the symbiosis by mediating cell-cell interactions.

Late Larval Development and Onset of Symbiosis in the Scleractinian Coral Fungia scutaria.

Many corals that harbor symbiotic algae (zooxanthellae) produce offspring that initially lack zooxanthellae. This study examined late larval development and the acquisition of zooxanthellae in the scleractinian coral Fungia scutaria, which produces planula larvae that lack zooxanthellae. Larvae reared under laboratory conditions developed the ability to feed 3 days after fertilization; feeding behavior was stimulated by homogenized Artemia. Larvae began to settle and metamorphose 5 days after fertilization. In laboratory experiments, larvae acquired experimentally added zooxanthellae by ingesting them while feeding. Zooxanthellae entered the gastric cavity and were phagocytosed by endodermal cells. As early as 1 h after feeding, zooxanthellae were observed in both endodermal and ectodermal cells. Larvae were able to form an association with three genetically distinct strains of zooxanthellae. Both zooxanthellate and azooxanthellate larvae underwent metamorphosis, and azooxanthellate polyps were able to acquire zooxanthellae from the environment. Preliminary evidence suggests that the onset of symbiosis may influence larval development; in one study symbiotic larvae settled earlier than aposymbiotic larvae. Protein profiles of eggs and larvae throughout development revealed a putative yolk protein doublet that was abundant in eggs and 1-day-old larvae and was absent by day 6. This study is the first to examine the onset of symbiosis between a motile cnidarian host and its algal symbiont.

Fear of falling revisited.

OBJECTIVE: To evaluate the measurement properties of an expanded version of the Falls Efficacy Scale (FES)-a measure of fear of falling. The original FES measures fear on almost exclusively indoor activities, which may limit the usefulness of the scale in identifying early stages of fear of falling in active community-dwelling older people. DESIGN: Two-group convenience sample. SETTING: An outpatient referral clinic, and community-dwelling older people recruited from various sources. SUBJECTS: A volunteer sample of 111 healthy community-dwelling elderly (mean age 74.0 years) and a sample of 68 older people referred to a Falls and Balance Clinic (FBC) (mean age 79.2). Twenty-one subjects (9 healthy elderly and 12 FBC patients) were tested twice 1 week apart to investigate retest reliability. MAIN OUTCOME MEASURES: A 14-activity questionnaire (the Modified Falls Efficacy Scale [MFES]) was used that incorporated the original 10-activity FES and four additional activities. Falls efficacy was rated on a 10-point visual analogue scale for each activity. RESULTS AND CONCLUSIONS: The MFES demonstrated high internal consistency (Cronbach's alpha.95) and less skew than the original FES (-2.4 and -3.3, respectively). Factor analysis of the MFES revealed two factors accounting for 75% of the sample variance, grouping into an "indoor type activity" factor and an "outdoor type activity" factor. Retest reliability for the MFES was high (intraclass correlation coefficients = .93). Significant differences were evident between the FBC group and the healthy older group on all items of the MFES and on the total MFES score (p < .05). On the basis of these preliminary findings, the MFES appears to be a reliable and valid measure of falls self-efficacy, and could be a useful addition in the comprehensive assessment of older people with balance disturbance or falls.

A falls and balance clinic for the elderly.

A Falls and Balance Clinic for the elderly has been developed with the aim of identifying those at risk of subsequent falls, and recommending intervention strategies to reduce this risk. The physical and functional status of 149 clients referred to the Clinic are reported. Neurological and musculoskeletal pathologies were identified as the cause of falls in the majority of clients. Measures of gait velocity and stride length, and ability to stand on one leg were markedly reduced compared with normative data for healthy elderly. A number of other measures of balance, strength, and function are also reported. The results of a questionnaire about home environment indicated that only 28% of those referred had been assessed in their own home in the previous year. Home environment was considered a potential risk in a further 28% of clients and a home visit was instituted in these cases. Other intervention strategies included referral for further investigations (36%), Day Hospital (33%), provision of a home program of balance or strengthening exercises (27%), and medication change (15%). Issues related to the establishment, operation and long term evaluation of the effectiveness of a specialist Falls and Balance Clinic for the elderly are discussed.

The effects of lansoprazole, 30 or 60 mg daily, on intragastric pH and on endocrine function in healthy volunteers.

Seven days of dosing with either 30 mg or 60 mg of lansoprazole were compared with placebo in a double-blind, randomized, three-way cross-over study in 12 male healthy volunteers. Twenty-four-hour intragastric pH was measured after 7 days of dosing with each regimen, as well as 3 and 7 days after the end of dosing. During dosing with placebo, intragastric pH was above 4 for a median of 51 minutes. pH values were significantly raised to above 4 for 8.45 and 8.33 hours on Day 7 of dosing with lansoprazole 30 and 60 mg, respectively, but returned to normal by the third day after stopping dosing. No clinically relevant influence on endocrine function (serum concentrations of insulin, aldosterone, testosterone, parathormone, glucagon, T3, T4, TSH, LH, FSH, STH, prolactin, circadian cortisol profile, ACTH test) was observed. No serious adverse clinical or laboratory events were noted.

Lansoprazole versus famotidine: efficacy and tolerance in the acute management of duodenal ulceration.

Lansoprazole (AG 1749/CG 4801) is an inhibitor of gastric acid secretion by blocking H+,K(+)-ATPase. In this 2:1 randomized, double-blind, multicentre trial lansoprazole 30 mg am was compared to 40 mg famotidine nocte in 264 out-patients suffering from uncomplicated duodenal ulcer. After 2 weeks of treatment ulcer healing was confirmed endoscopically in a significantly higher proportion (P = 0.027) of patients treated with lansoprazole (94/174 = 54.0%) compared to patients receiving famotidine (35/90 = 38.9%). Cumulative healing rates after 4 weeks were 91.4% for the lansoprazole group and 83.3% for the famotidine group (P = 0.065). Pain relief and decrease of concomitant antacid consumption during treatment were comparable in both groups. Both compounds were well tolerated. Rates of recurrent duodenal ulcer in the 6 months after trial treatment were 45/158 (28.5%) after lansoprazole, and 18/69 (26.1%) after famotidine.

A cross-over study of oral and intravenous administration of theophylline in male volunteers. Absolute bioavailability of theophylline tablets.

Seven normal male volunteers each received an injection of 208 mg theophylline anhydride for 4 min, and a 185-mg single oral dose theophylline anhydride (Broncho-Afonilum) on separate occasions. The concentrations of unchanged theophylline was analyzed over a 24-h period in plasma and a 48-h period in urine after both dosage forms. A 2-compartment model was required to describe the intravenous plasma concentration-time course in all seven subjects. A 1-compartment model sufficed to account for the decay of the oral plasma concentrations in all subjects. The mean plasma t1/2 after i.v. dosing was 6.5 h (3.5-9 h), and the mean plasma t1/2 after oral doses was 5.7 h (3-12 h). The calculated total of 23% (11-70%) of the intravenous dose was excreted in urine, and the mean total excretion after the oral dose was 12% (7-19%). The absolute bioavailability of oral theophylline was 94 +/- 20%

A crossover study after oral and intravenous administration of theophylline in male volunteers (absolute bioavailability of Afonilum tablets).

Seven normal male volunteers received an injection of 208 mg theophylline-anhydride (Solosin) each for 4 min, and a 225-mg single oral dose (Afonilum) on separate occasions. The concentration of unchanged theophylline was analyzed over a 24-h period in plasma and over a 48-h period in urine, after both dosage forms. A two-compartment model was required to describe the i.v. plasma concentration-time course in all seven subjects. A one-compartment model sufficed to account for the decay of the oral plasma concentrations in five of the subjects. The mean plasma t 1/2 after i.v. dosing was 6.5 h (3.5-9 h), and the mean plasma t 1/2 after oral doses was 5.9 h (3-8.3 h). The calculated mean total of 23% (11-70%) of the i.v. dose was excreted in the urine, and the mean total excretion after the oral dose was 15% (7-44%). The absolute bioavailability of Afonilum was 108 +/- 11%.

Pharmacokinetics of low molecular (monovalent) dextran (Dx 1) in volunteers.

Dextran-induced side effects were attributed to preformed antibodies that cross-react with dextran. These antibodies can be blocked by monovalent haptens (dextrans 1) in animals. Dextran 1 is also well tolerated in humans. Plasma levels and renal excretion of monovalent dextran (dextran 1, molecular weight around 1,000) were measured in five volunteers after intravenous administration of 20 ml 15% dextran 1 (3 g), and in one volunteer after intravenous administration of 50 ml (7.5 g). Measurements could be satisfactorily described by a two-compartment open model with elimination from the central compartment only: mean half-life for the beta phase of 1.9 h, a mean cumulative asymptotic elimination of dextran in the urine of 75%, and a mean renal clearance of 137 ml/min with a mean total clearance of 187 ml/min were estimated. The terminal half-life of dextran 60 (Macrodex) was 24 h (median). In volunteers with dextran antibodies no anaphylactoid symptoms were observed after Dx 1. In conclusion, the intravenous preinjection of dextran 1, owing to its pharmacokinetic behavior, should prevent antibody-mediated side effects after infusions with clinical dextrans (e.g., Macrodex, Rheomacrodex).

[Prevention of antibody mediated dextran side effects by hapten inhibition in dogs (author's transl)]. / Verhinderung Antikörper-bedingter Dextran-Nebenwirkungen durch Hapten-Hemmung mit Dextran 1 am Hund.

The effectiveness of low molecular dextran (dextran 1) in preventing antibody mediated dextran side effects (hapten inhibition) was investigated in immunized dogs. After injection of clinical dextran (dextran 60) a rapid fall of systolic blood pressure and a decrease of leucocytes, thrombocytes and haemolytic complement activity was noted. The protective effect of a dextran 1 preinjection was clearly demonstrated; in contrast the built-in hapten injection (dextran + hapten together) was ineffective.

[Activation of the complement system with clinical dextran (Macrodex 6%) in 10 patients in a cross-over design against 0,9%-NaCl]. / Untersuchung zur Aktivierung des Komplementsystems durch klinisches Dextran (Macrodex 6%) an 10 Probanden im Cross-over-Design gegen 0,9%-NaCl.

The possibility of activation of the complement system with clinical Dextran 60 (500 ml Macrodex 6% infusion) as compaired with 500 ml 0.9% NaCl was investigated within a one week interval in 10 volunteers of both sexes in a cross-over design. Anaphylactoid symptoms, which would indicate a release of biological activities, were not induced by Dextran 60 or 0.9% NaCl. Neither could a change in blood pressure, heart rate, in the whole hemolytic complement activity nor in the leucocyte and thrombocyte count be determined. Therefore, a clinical relevant activation of the complement system in normal persons by Macrodex 6% can not be assumed.

[Hapten inhibition: inhibition of antibody dependent dextran side effects by means of low molecular dextran (dextran 1) as a monovalent hapten]. / Hapten-Hemmung: Verhinderung antikörperbedingter Dextran-Nebenwirkungen durch niedermolekulares Dextran (Dextran 1) als monovalentes Hapten.

Dextran-induced anaphylactoid reactions can be attributed to preformed circulating antibodies cross-reacting with dextrans. Therefore, after intravenous administration of clinical dextran, formation of immune complexes with resulting complement activation and secondary formation and/or release of mediators with clinical symptoms of an immune complex or aggregate anaphylaxis can occur (type-III immune reaction). Antigen antibody reactions can be prevented if the antigen-binding sites of the antibodies are specifically blocked by monovalent haptens (low molecular non-immunogenic antigens). The effectiveness of this principle of 'hapten inhibition' could be proven in vitro as well as in animal experiments. In volunteer studies intravenous administration of the low molecular dextran in a 15% solution (Dextran 1), with a molecular weight around 1,000 functioning as monovalent hapten, was well tolerated.