Substrate usage and its regulation during flight and swimming in the backswimmer, Notonecta glauca

Abstract.  The metabolites that are generally used by insects during exercise are present in quite different concentrations in the haemolymph of the backswimmer Notonecta glauca L. Lipids are most abundant (between 10 and 20 mg/mL), whereas carbohydrates (2–3 mg/mL) and proline (approximately 1 mg/mL) are at very low concentrations. Injection of an extract of conspecific corpora cardiaca causes pronounced hyperlipaemia in the backswimmer. A neuropeptide with the same effect was isolated from the corpora cardiaca in a single high-performance liquid chromatography (HPLC) step; the primary sequence was deduced from mass spectrometric measurements (matrix-assisted laser desorption/ionization-time of flight and electrospray quadrupol time-of-flight mass spectrometry) of whole corpora cardiaca, and the mass was confirmed in the HPLC fraction that had adipokinetic activity. The biologically active octapeptide has the sequence pGlu-Val-Asn-Phe-Ser-Pro-Ser-Trp amide, which was characterized previously from the corpora cardiaca of the Emperor dragonfly, Anax imperator , and denoted Anaim-adipokinetic hormone (AKH). The synthetic Anaim-AKH peptide causes lipid mobilization when injected at a dose of 1 pmol into N. glauca . When other synthetic AKH members that occur in Hemiptera are injected into N. glauca at the same dose, the hyperlipaemic responses are significantly lower than after injection of Anaim-AKH. Because only lipids increase upon activity, such as continuous swimming for 1 h or during a 1-h rest period after a 3-min flight episode in the laboratory, it is assumed that Anaim-AKH serves as a true adipokinetic hormone in the backswimmer during bouts of natural swimming and flight.     

Influence of aminergic and peptidergic substances on heart beat frequency in the stick insect Carausius morosus (Insecta,Phasmatodea)

The dorsal heart of the Indian stick insect, Carausius morosus, is responsible for the anterograde flow of hemolymph to the aorta and into the body cavity. The contraction frequency of the insect heart is known to be influenced by several substances of neural source. Here, a semi‐exposed heart assay was employed to study the effect of an aminergic substance (octopamine) and three neuropeptides (C. morosus hypertrehalosemic hormone [Carmo‐HrTH], crustacean cardioactive peptide [CCAP], and proctolin) on heart contraction. The contraction frequency was measured as beats per minute in adults ligated between the head and the prothorax. All three investigated neuropeptides had a stimulatory effect on heart contraction that lasted approximately 6 min, after which the normal heart beat rate was restored. Proctolin and CCAP stimulated the rate of heart beat also in unligated stick insects, whereas Carmo‐HrTH was active only in ligated insects. The latter could suggest that when the stick insect is not ligated, a competing substance may be released from the head of C. morosus; the competing substance is, apparently, not physiologically active but it binds or blocks access to the receptor of Carmo‐HrTH‐II, thereby rendering the HrTH peptide “not active.” In ligated stick insects, 6.7 × 10^?8 M Carmo‐HrTH‐II significantly increased the heart beat rate; higher doses resulted in no further increase, suggesting the saturation of the HrTH receptor. Octopamine inhibited the rate at which the heart contracted in a dose‐dependent manner; inhibition was achieved with 10^?4 M of octopamine.     

Synthesis and assay of structural intermediates of crustacean pigment-dispersing hormones (α and β-PDH)

Summary

The two characterized crustacean pigment-dispersing hormones (α-PDH; β-PDH) are octadecapeptides which differ in primary structure at six positions. Assays for melanophore pigment-dispersing activity showed β-PDH to be 21-fold more potent than α-PDH. In an effort to explain the difference in potencies between the two PDHs, we synthesized and purified six analogs of α-PDH (Leu^4?, Leu^11?, Lys^13?, Asn^16?, Asp^17?, and Glu³, Leu^4? α-PDH) in which the amino acid residues of α-PDH were substituted with those of β-PDH. Four analogs (Leu^11?, Lys^13?, Asn^16?, and Asp^17? α-PDH) possessed melanophore-dispersing activity equivalent to α-PDH. Leu^4? α-PDH and Glu³, Leu^4? α-PDH were 2.4? and 4-fold more potent than α-PDH, respectively. Glu^3-α-PDH was 3.3-fold more potent than α-PDH (Jorenby et al., 1987). These results suggest that the 21-fold increase in activity of β-PDH over α-PDH is due to an interactive effect of two or more substitutions rather than from the product of the effects brought about by individual substitutions.     

Influences of photoperiod and alternate days of feeding on plasma growth hormone and thyroid hormone levels in juvenile rainbow trout

The diurnal patterns of plasma growth hormone (GH), thyroid hormone and cortisol concentrations in rainbow trout Oncorhynchus mykiss held under three photoperiod (L : D) regimes (6 : 18, 12 : 12 and 18 : 6), and fed either daily (DF) or on alternate days (ADF) with 2·0% body mass per day of a commercial trout diet were determined. The ADF groups had reduced total mass gain and specific growth rates compared with DF fish, but photoperiod had no affect on growth for either of the feeding regime groups. In the ADF groups, the mean 24 h plasma thyroxine (T₄) and triiodothyronine (T₃) concentrations were significantly lower, both on days of feeding and days of fasting, than in DF fish held under all three photoperiod regimes, but for GH, only the 18L : 6D DF group was higher than the comparable ADF groups. There were no significant differences in mean 24 h plasma cortisol concentrations of DF and ADF groups. Diurnal patterns of plasma GH, cortisol, T₄ and T₃ were found in DF fish held under all three photoperiod regimes. Increases in plasma cortisol changes were associated with the onset of the light phase; elevations in plasma GH and T₄ concentrations were more closely associated with clock time, regardless of photoperiod; increases in plasma T₃ concentrations were strongly associated with time of feeding. In ADF groups, these diurnal changes in plasma GH, T₄ and T₃ concentrations were suppressed for both the fed and fast days, and plasma cortisol concentrations were suppressed on the fasting day. The observations are discussed in terms of the proposed anabolic, catabolic and growth regulating roles of these hormones in different growth and metabolic modifying situations in teleosts.     

Synthesis and characterization of novel biotinylated carboxyl-terminal parathyroid hormone peptides that specifically crosslink to the CPTH-receptor

Parathyroid hormone (PTH) regulates calcium, phosphorous and skeletal homeostasis via interaction with the G protein-coupled PTH/PTHrP receptor, which is fully activated by the amino-terminal 34 amino-acid portion of the hormone. Recent evidence points to the existence of another class of receptors for PTH that recognize the carboxyl (C)-terminal region of intact PTH (1–84) (CPTHRs) and are highly expressed by osteocytes. Here we report the synthesis and characterization of two novel bifunctional CPTH ligands that include benzoylphenylalanine (Bpa) substitutions near their amino-termini and carboxyl-terminal biotin moieties, as well as a tyrosine³⁴ substitution to enable radioiodination. These peptides are shown to bind to CPTHRs with affinity similar to that of PTH (1–84) and to be specifically and covalently crosslinked to CPTHRs upon exposure to ultraviolet light. Crosslinking to osteocytes or osteoblastic cells generates complexes of 80 and 220 kDa, of which the larger form represents an aggregate that can be resolved into the 80 kDa. The crosslinked products can be further purified using immunoaffinity and avidin-based affinity procedures. While the molecular structure of the CPTHR(s) remains undefined, these bifunctional ligands represent powerful new tools for use in isolating and characterizing CPTHR protein(s).     

Caprine luteinizing hormone isoforms during the follicular phase and anestrus

The relative proportion of the circulating luteinizing hormone isoforms in goats during follicular phase (pre-ovulatory peak; F) and anestrus (A) was investigated. Estrus was synchronized in six goats with a prostaglandin analogue. After estrus was detected, blood samples were taken at 1 h intervals for 24 h. Four anestrous goats received 100 μg i.v. of GnRH and blood samples were collected every 15 min for 5 h. Samples with the greatest LH concentration in follicular phase and after GnRH administration (anestrus) were analyzed by chromatofocusing and eluted with a pH gradient from 10.5 to 3.5. For quantification purposes eluted LH was grouped into basic (pH ≥ 7.5), neutral (pH 7.4–6.5) and acidic isoforms (pH ≤ 6.4) as well as by pH unit. In both physiological conditions (PC), basic and acidic isoforms were greater than the neutral. With this grouping criteria, there was an interaction between PC and pH group, with the proportion of neutral isoforms being greater (p < 0.05) in A (12.0 ± 0.8%) as compared with F (5 ± 2%). Analysis by pH unit showed a very basic group of eluted isoforms (pH ≥ 10), which amounted to a percentage of 6.0 ± 0.4% of the total observed during A, and 3 ± 1% during F (p < 0.05). Predominant isoforms in A eluted in the pH range 9.99–9.0 (42 ± 3%) as compared to 7 ± 3% (p < 0.01) in that pH range in F. In contrast, the predominant isoforms in F eluted in the pH range 8.99–8.0, representing 55 ± 8%, while in A the proportion was 11 ± 2% (p < 0.01). Isoforms eluted at the pH range 7.9–7 represented a significantly greater proportion during A (5.0 ± 0.6%) as compared with F (3 ± 1%). This is the first report on goat LH circulating isoforms. During A the LH isoforms secreted by the pituitary are more basic than during F.     

Calcium influx enhances neuropeptide activation of ecdysteroid hormone production by mosquito ovaries

A critical step in mosquito reproduction is the ingestion of a blood meal from a vertebrate host. In mosquitoes like Aedes aegypti, blood feeding stimulates the release of ovary ecdysteroidogenic hormone (OEH) and insulin-like peptide 3 (ILP3). This induces the ovaries to produce ecdysteroid hormone (ECD), which then drives egg maturation. In many immature insects, prothoracicotropic hormone (PTTH) stimulates the prothoracic glands to produce ECD that directs molting and metamorphosis. The receptors for OEH, ILP3 and PTTH are different receptor tyrosine kinases with OEH and ILP3 signaling converging downstream in the insulin pathway and PTTH activating the mitogen-activated protein kinase pathway. Calcium (Ca²⁺) flux and cAMP have also been implicated in PTTH signaling, but the role of Ca²⁺ in OEH, ILP3, and cAMP signaling in ovaries is unknown. Here, we assessed whether Ca²⁺ flux affects OEH, ILP3, and cAMP activity in A. aegypti ovaries and also asked whether PTTH stimulated ovaries to produce ECD. Results indicated that Ca²⁺ flux enhanced but was not essential for OEH or ILP3 activity, whereas cAMP signaling was dependent on Ca²⁺ flux. Recombinant PTTH from Bombyx mori fully activated ECD production by B. mori PTGs, but exhibited no activity toward A. aegypti ovaries. Recombinant PTTH from A. aegypti also failed to stimulate either B. mori PTGs or A. aegypti ovaries to produce ECD. We discuss the implications of these results in the context of mosquito reproduction and ECD biosynthesis by insects generally.     

Endocrine effects of social stimuli on maturing queens of the dampwood termite Zootermopsis angusticollis

Abstract.  The reproductive physiology of social insects is often highly responsive to social stimuli from nestmates, but the mechanisms underlying this sensitivity are usually poorly understood. The effect of varied social conditions on the endocrinology and ovarian maturation in maturing female primary reproductives of the dampwood termite Zootermopsis angusticollis Hagen is studied to better understand their developmental responses to conspecifics. Newly dealate queens are paired with a reproductive male, with another maturing female, or are allowed to mature in isolation. Developmental responses are tracked by monitoring rates of juvenile hormone (JH) production by corpora allata in vitro , haemolymph ecdysteroids titres and ovarian development after 5, 10, 15 and 30 days of maturation. Significant declines in rates of JH production rates are observed by days 5, 10 and 30, respectively, for females paired with males, left to mature in isolation or paired with a female. Ecdysteroid titres increase by day 5 and stay elevated in females nesting with males. By contrast, for CA from females that are isolated or paired with another female, the titres decline and stay low until at least day 15, but increase significantly by day 30. These hormonal differences correspond to differential rates of physiological maturation, with more rapid ovarian development occurring in females paired with a male, than in those nesting alone or with another female. The results suggest that JH and ecdysteroids modulate the ovarian response of alates to stimuli from nestmates during this period of maturation, and that JH and ecdysteroid titres may be regulated independently during this period of development.     

Movement of proteins across the digestive system of the tobacco budworm, Heliothis virescens

Bovine serum albumin (BSA) and anti‐BSA polyclonal antibody were used as model polypeptides to examine the movement of foreign proteins across the insect digestive system and their accumulation in hemolymph of fourth stadium tobacco budworms, Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae). Hydrateable meal pads were developed in these studies as a method for easily introducing compounds into the insect digestive system. When insects were allowed to feed continuously on hydrated meal pads containing 0.8 mg of anti‐BSA per gram diet, the level of antibody found in hemolymph was 2.4 ± 0.1 and 3.4 ± 0.1 µg ml^?1 (average  1 SEM) after 8 and 16 h, respectively, as determined by enzyme‐linked immunosorbant assay (ELISA). Continuous feeding on hydrated meal pads containing the same concentration of BSA produced hemolymph concentrations of 1.5 ± 0.1 and 1.6 ± 0.1 µg ml^?1 hemolymph at 8 and 16 h, respectively. Western blot analyses demonstrated that BSA and anti‐BSA both retained their primary and multimeric structure and that anti‐BSA maintained its antigenic activity in the meal pads and after movement from meal pads into the hemolymph. When 1 µg of anti‐BSA or BSA was injected into the hemocoel of fourth instars, the concentrations decreased with time and 120 min after injection were 20% and 0.6% of the original concentration, respectively. When added at the same concentration to plasma in vitro, the decrease was 81.5% and 57.5%, respectively, at 2 h. The accumulation of native anti‐BSA and BSA protein in insect hemolymph is the result of their rate of movement across the gut and their rate of turnover in hemolymph. Movement of anti‐BSA and BSA across the digestive system was also noted in Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), Acheta domesticus (L.) (Orthoptera: Gryllidae), and Gromphadorhina portentosa (Schaum) (Blattaria: Blattellidae). Anti‐BSA and BSA were not detected in the hemolymph of Manduca sexta (L.) (Lepidoptera: Sphingidae) after feeding.     

Calcitonin gene-related peptide-mediated cardioprotection of postconditioning in isolated rat hearts

Previous studies have demonstrated that endogenous calcitonin gene-related peptide (CGRP) plays an important role in mediation of ischemic preconditioning. In the present study, we tested whether CGRP is also involved in mediation of the protective effects of postconditioning in isolated rat hearts. Sixty minutes of left coronary artery occlusion and followed by 60 min of reperfusion caused a significant decrease in cardiac function and a significant increase in creatine kinase (CK) release and infarct size. Postconditioning with three cycles of 1-min ischemia and 1-min reperfusion produced a marked improvement of cardiac function and decreased CK release and infarct size, concomitantly with an increase in the release of CGRP release in coronary effluent. However, the cardioprotection afforded by postconditioning was abolished by CGRP 8-37 (10^− 7 M), a selective CGRP receptor antagonist, or pretreatment with capsaicin (50 mg/kg, s.c.), which depletes transmitters in sensory nerves. Exogenous CGRP (5 × 10^− 9 M) administration of CGRP reappeared postconditioning-like cardioprotection in the rats pretreated with capsaicin. These results suggest that the protective effects of ischemic postconditioning are related to stimulation of endogenous CGRP release in rat hearts.     

Peptide amidation: Production of peptide hormonesin vivo andin vitro

Over half of all biologically active peptides and peptide hormones are α-amidated at their C-terminus, which is essential for their full biological activities. Amidation is accomplished through the sequential reaction of the two enzymes encoded by the single bifunctional, peptidylglycine α-amidating monooxygenase (PAM or an α-amidating enzyme). PAM catalyzes the formation of a peptide amide from peptide precursors that include a C-terminal glycine, and requires copper, molecular oxygen, and ascorbate. PAM is the only enzyme that produces peptide amidesin vivo. However, various strategies utilizing PAM, carboxypeptidase-Y enzymes, and chemical synthesis have been developed for producing peptide amidesin vitro. The growing need and importance of peptide amide drugs has highlighted the necessity for an efficientin vitro amidating system for industrial application. In recent years, recombinant systems for enzymatic amidation have received growing attention for the production of peptide hormones, like calcitonin and oxytocin. This review presents the current situation regarding amidation, with a special emphasis on the industrial production of peptide hormones.     

Cell-penetrating peptides as antifungals towards Malassezia sympodialis

Aim: To determine whether different antimicrobial peptides (AMPs) and cell‐penetrating peptides (CPPs) are able to inhibit the growth of the commensal yeast Malassezia sympodialis, which can act as a trigger factor in different skin disorders, such as atopic eczema (AE), seborrhoeic eczema (SE) and dandruff. Methods and results: The antifungal activity of 21 different AMPs and CPPs was investigated by microdilution assay and plate counting to determine the number of colony forming units. Five CPPs and one AMP showed fungicidal activity at submicromolar concentrations. Importantly, no membrane damage on human keratinocytes was detected after peptide treatment. Conclusions: Several CPPs, while being nontoxic to mammalian cells, possess growth inhibitory activity on the very stringent yeast M. sympodialis. Significance and impact of study: Our findings that five CPPs and one AMP that are harmless towards mammalian cells act as antifungal agents against M. sympodialis opens up the possibility to use these in the treatment for AE, SE and dandruff. To our knowledge, this is the first time peptides have been identified as antifungal agents against M. sympodialis. Further studies to elucidate the mechanism are warranted.     

Synthetic glycosylation of peptides using unprotected saccharide β-glycosylamines

Glycopeptides can be valuable tools in determining the influence of carbohydrate moieties on the intrinsic properties of glycoproteins. However, glycopeptides of sufficient quantity and purity are as yet not readily available from biological sources. The chemical coupling of a -glycosylamino group of an unprotected carbohydrate with an activated aspartic acid residue of an unprotected peptide is a simple method for synthesizing asparagine-linked glycopeptides. In this report we demonstrate that the use of this method is not restricted to -glycosylamines of simple monosaccharides or short aspartic acid-containing pentapeptides. This is illustrated by the syntheses of several glycopentapeptides containingN,N-diacetylchitobiose, a glutamine-linked glycopentapeptide containing a biantennary complex oligosaccharide, and glycosylated variants of two analogs of a polypeptide hormone, atriopeptin, containingN,N-diacetylchitobiose.Abbreviations Ac acetyl - Bzl benzyl - DMF dimethylformamide - Fmoc 9-fluorenylmethoxycarbonyl - Fuc fucose - Gal galactose - GlcNAc N-acetylglucosamine - HBTU O-benzotriazol-1-yl-N,N,N,N-tetramethyluroniumhexa-fluorophosphate - HOBt 1-hydroxybenzotriazole - Man mannose - m/z mass/charge - NMR nuclear magnetic resonance - Xyl xylose - Z benzyloxycarbonyl; unless otherwise specified, amino acids are abbreviated using their one-letter codes.     

Growth hormone secretion is diminished and tightly controlled in humans enriched for familial longevity

Reduced growth hormone (GH) signaling has been consistently associated with increased health and lifespan in various mouse models. Here, we assessed GH secretion and its control in relation with human familial longevity. We frequently sampled blood over 24 h in 19 middle‐aged offspring of long‐living families from the Leiden Longevity Study together with 18 of their partners as controls. Circulating GH concentrations were measured every 10 min and insulin‐like growth factor 1 (IGF‐1) and insulin‐like growth factor binding protein 3 (IGFBP3) every 4 h. Using deconvolution analysis, we found that 24‐h total GH secretion was 28% lower (P = 0.04) in offspring [172 (128–216) mU L^?1] compared with controls [238 (193–284) mU L^?1]. We used approximate entropy (ApEn) to quantify the strength of feedback/feedforward control of GH secretion. ApEn was lower (P = 0.001) in offspring [0.45 (0.39–0.53)] compared with controls [0.66 (0.56–0.77)], indicating tighter control of GH secretion. No significant differences were observed in circulating levels of IGF‐1 and IGFBP3 between offspring and controls. In conclusion, GH secretion in human familial longevity is characterized by diminished secretion rate and more tight control. These data imply that the highly conserved GH signaling pathway, which has been linked to longevity in animal models, is also associated with human longevity.     

Capsaicin-evoked bradycardia in anesthetized guinea pigs is mediated by endogenous tachykinins

The present study was done to characterize the effects of endogenous tachykinins on heart rate in urethane-anesthetized guinea pigs. Intravenous injection of capsaicin (32 nmol/kg) was used to evoke release of tachykinins and calcitonin gene-related peptide (CGRP) from cardiac sensory nerve fibers. Such injections caused a brief decrease in heart rate (− 37 ± 7 beats/min, n = 6) that was followed by a more prolonged increase (+ 44 ± 10 beats/min). Blood pressure was lowered by − 11 ± 2 mmHg. Bilateral vagotomy did not affect the chronotropic or depressor responses to capsaicin, but atropine (1 µmol/kg) nearly abolished the bradycardic response (− 8 ± 3 beats/min, n = 7). Combined blockade of NK₂ and NK₃ receptors, with SR48968 and SR14801 respectively, also caused a significant reduction of capsaicin-evoked bradycardia (− 14 ± 3 beats/min, n = 4) but did not affect bradycardia evoked by vagal nerve stimulation. Blockade of CGRP receptors eliminated capsaicin-evoked tachycardia and prolonged the capsaicin-evoked bradycardia. These findings suggest that capsaicin-evoked bradycardia in the anesthetized guinea pig is mediated by tachykinins that stimulate cardiac cholinergic neurons. This effect appears to be truncated by the positive chronotropic action of CGRP that is also released from cardiac afferents by capsaicin.     

High microbial activity on glaciers: importance to the global carbon cycle

Cryoconite holes, which can cover 0.1–10% of the surface area of glaciers, are small, water-filled depressions (typically <1 m in diameter and usually <0.5 m deep) that form on the surface of glaciers when solar-heated inorganic and organic debris melts into the ice. Recent studies show that cryoconites are colonized by a diverse range of microorganisms, including viruses, bacteria and algae. Whether microbial communities on the surface of glaciers are actively influencing biogeochemical cycles or are just present in a dormant state has been a matter of debate for long time. Here, we report primary production and community respiration of cryoconite holes upon glaciers in Svalbard, Greenland and the European Alps. Microbial activity in cryoconite holes is high despite maximum temperatures seldom exceeding 0.1 °C. In situ primary production and respiration in cryoconites during the summer is often comparable with that found in soils in warmer and nutrient richer regions. Considering only glacier areas outside Antarctica and a conservative average cryoconite distribution on glacial surfaces, we found that on a global basis cryoconite holes have the potential to fix as much as 64 Gg of carbon per year (i.e. 98 Gg of photosynthesis minus 34 Gg of community respiration). Most lakes and rivers are generally considered as heterotrophic systems, but our results suggest that glaciers, which contain 75% of the freshwater of the planet, are largely autotrophic systems.     

High host specificity of obligate ectoparasites

Abstract.  1. Host specificity is the degree to which a parasite species occurs in association with a host species.
2. The degree to which obligate ectoparasites are host specific has been debated, but effects of sampling contamination were usually not addressed. Data from a controlled mammal–ectoparasite survey were used to assess host specificity of an obligate group of ectoparasites – streblid bat flies.
3. Host–parasite associations were categorised as primary or non-primary. Non-primary host associations were evaluated against primary associations via proportional comparison.
4. Results indicate that host specificity was high, exceeding previous reports. Natural host transfers were rare.
5. Non-primary host associations were almost completely explained by disturbance transfers during sampling of the host or by contamination upon sampling the parasite. These conclusions likely hold for other taxa of obligate parasites.     

Amoxicillin/tetracycline combinations are inadequate as alternative therapies for Helicobacter pylori infection

Background. Triple therapy with proton pump inhibitors or ranitidine bismuth citrate, clarithromycin and either amoxicillin or nitroimidazole derivatives are the present gold standards for cure of Helicobacter pylori infection. However, primary resistance to either clarithromycin or nitroimidazole derivatives is increasing and alternative therapies are needed. Aim. To determine the efficacy and safety of three regimens consisting of amoxicillin and tetracycline or doxycycline combined with either lansoprazole or ranitidine bismuth citrate. Methods. Two hundred and seventy H. pylori infected patients were randomly given one of the following treatments: amoxicillin 1 g twice a day (b.i.d.) plus tetracycline 500 mg four times a day (q.i.d.) with either lansoprazole 30 mg b.i.d. (group LAT) or ranitidine bismuth citrate 400 mg b.i.d. (group RBCAT) for 7 days and amoxicillin 1 g b.i.d. plus doxycycline 100 mg b.i.d. and lansoprazole 30 mg b.i.d. for 14 days (group LAD). Eradication rate was assessed by UBT at 4–6 weeks after therapy. Results. The three groups (LAT, RBCAT, and LAD) of patients achieved eradication rates of 35% (25–45), 20% (12–29) and 36% (25–46), respectively, on intention‐to‐treat analysis. Patient compliance was optimal and side‐effects minimal in all three groups. Conclusions. Although the amoxicillin/tetracycline combination is attractive (inexpensive, safe, and with low primary resistance rate), it can not be recommended for H. pylori eradication.