Abrupt decrease in serum testosterone levels after an oral glucose load in men: implications for screening for hypogonadism.

OBJECTIVE: This study examines the physiological impact of a glucose load on serum testosterone (T) levels in men with varying glucose tolerance (GT). DESIGN: Cross-sectional study. PATIENTS AND METHODS: 74 men (19-74 years, mean 51·4 ± 1·4 years) underwent a standard 75-g oral glucose tolerance test with blood sampling at 0, 30, 60, 90 and 120 min. Fasting serum glucose, insulin, total T (and calculated free T), LH, SHBG, leptin and cortisol were measured. RESULTS: 57% of the men had normal GT, 30% had impaired GT and 13% had newly diagnosed type 2 diabetes. Glucose ingestion was associated with a 25% decrease in mean T levels (delta = -4·2 ± 0·3 nm, P < 0·0001). T levels remained suppressed at 120 min compared with baseline (13·7 ± 0·6 vs 16·5 ± 0·7 nm, P < 0·0001) and did not differ across GT or BMI. Of the 66 men with normal T levels at baseline, 10 (15%) had levels that decreased to the hypogonadal range (<9·7 nm) at one or more time points. SHBG, LH and cortisol levels were unchanged. Leptin levels decreased from baseline at all time points (P < 0·0001). CONCLUSIONS: Glucose ingestion induces a significant reduction in total and free T levels in men, which is similar across the spectrum of glucose tolerance. This decrease in T appears to be because of a direct testicular defect, but the absence of compensatory changes in LH suggests an additional central component. Men found to have low nonfasting T levels should be re-evaluated in the fasting state.

The lack of effect of insulin on luteinizing hormone pulsatility in healthy male volunteers provides evidence of a sexual dimorphism in the metabolic regulation of reproductive hormones.

BACKGROUND: The activity of the neuroendocrine reproductive axis is closely related to nutritional status. This link is particularly important in healthy women, in whom insulin is a positive signal for the reproductive system. In contrast, very little is known regarding this relation in men. OBJECTIVES: This study was designed to evaluate the effect of insulin on the reproductive axis of young male volunteers and to study the effect of short-term hypercaloric feeding on this modulation. DESIGN: The activity of the neuroendocrine reproductive axis was characterized by the pattern of endogenous luteinizing hormone (LH) secretion on the basis of frequent blood sampling protocols. The effect of insulin was tested by comparing the LH secretion pattern between a baseline study and a hyperinsulinemic euglycemic clamp. These studies were performed first in subjects fed a controlled isocaloric diet for 6 d (calculated as 1.5 times their resting metabolic rate) then in the same subjects fed a controlled hypercaloric diet in which 30% extra calories were provided as fat and fructose (3 g · kg(-1) · d(-1)) before undergoing identical protocols. Serum gonadotropins, sex steroids, glucose, insulin, ghrelin, and leptin concentrations were assessed, and the HOMA-IR was calculated. RESULTS: The LH secretion pattern was not affected by insulin or by hypercaloric feeding. Insulin decreased ghrelin and increased leptin concentrations but had no additional effect of hypercaloric feeding despite significantly lower HOMA-IR indexes. CONCLUSIONS: Our data indicate that neither insulin nor short-term hypercaloric feeding has any effect on the activity of the male reproductive axis. They also further support the association between ghrelin and insulin and glucose metabolism. This trial was registered at clinicaltrials.gov as NCT01058681.

Mutations in Plasmodium falciparum dihydrofolate reductase and dihydropteroate synthase of isolates from the Amazon Region of Brazil

Since the late 1970s pyrimethamine-sulfadoxine (PS; FansidarTM Hoffman-LaRoche, Basel) has been used as first line therapy for uncomplicated malaria in the Amazon basin. Unfortunately, resistance has developed over the last ten years in many regions of the Amazon and PS is no longer recommended for use in Brazil. In vitro resistance to pyrimethamine and cycloguanil (the active metabolite of proguanil) is caused by specific point mutations in Plasmodium falciparum dihydrofolate reductase (DHFR), and in vitro resistance to sulfadoxine has been associated with mutations in dihydropteroate synthase (DHPS). In association with a proguanil-sulfamethoxazole clinical trial in Brazil, we performed a nested mutation-specific polymerase chain reaction to measure the prevalence of DHFR mutations at codons 50, 51, 59, 108 and 164 and DHPS mutations at codons 436, 437, 540, 581 and 613 at three sites in the Brazilian Amazon. Samples from two isolated towns showed a high degree of homogeneity, with the DHFR Arg-50/Ile-51/Asn-108 and DHPS Gly-437/Glu-540/Gly-581 mutant genotype accounting for all infections in Peixoto de Azevedo (n = 15) and 60% of infections in Apiacás (n = 10), State of Mato Grosso. The remaining infections in Apiacás differed from this predominant genotype only by the addition of the Bolivia repeat at codon 30 and the Leu-164 mutation in DHFR. By contrast, 17 samples from Porto Velho, capital city of the State of Rondônia, with much in- and out-migration, showed a wide variety of DHFR and DHPS genotypes.

Novel loci for adiponectin levels and their influence on type 2 diabetes and metabolic traits: a multi-ethnic meta-analysis of 45,891 individuals.

Circulating levels of adiponectin, a hormone produced predominantly by adipocytes, are highly heritable and are inversely associated with type 2 diabetes mellitus (T2D) and other metabolic traits. We conducted a meta-analysis of genome-wide association studies in 39,883 individuals of European ancestry to identify genes associated with metabolic disease. We identified 8 novel loci associated with adiponectin levels and confirmed 2 previously reported loci (P = 4.5×10(-8)-1.2×10(-43)). Using a novel method to combine data across ethnicities (N = 4,232 African Americans, N = 1,776 Asians, and N = 29,347 Europeans), we identified two additional novel loci. Expression analyses of 436 human adipocyte samples revealed that mRNA levels of 18 genes at candidate regions were associated with adiponectin concentrations after accounting for multiple testing (p<3×10(-4)). We next developed a multi-SNP genotypic risk score to test the association of adiponectin decreasing risk alleles on metabolic traits and diseases using consortia-level meta-analytic data. This risk score was associated with increased risk of T2D (p = 4.3×10(-3), n = 22,044), increased triglycerides (p = 2.6×10(-14), n = 93,440), increased waist-to-hip ratio (p = 1.8×10(-5), n = 77,167), increased glucose two hours post oral glucose tolerance testing (p = 4.4×10(-3), n = 15,234), increased fasting insulin (p = 0.015, n = 48,238), but with lower in HDL-cholesterol concentrations (p = 4.5×10(-13), n = 96,748) and decreased BMI (p = 1.4×10(-4), n = 121,335). These findings identify novel genetic determinants of adiponectin levels, which, taken together, influence risk of T2D and markers of insulin resistance.

Gonadotropin-releasing hormone secretion from hypothalamic neurons: stimulation by insulin and potentiation by leptin.

Insulin and leptin are peripheral metabolic factors signaling the body needs in energy to the central nervous system. Because energy homeostasis and reproductive function are closely related phenomena, we investigated the respective roles played by insulin and leptin in the hypothalamic control of GnRH secretion. We observed that increasing circulating insulin levels, by performing hyperinsulinemic clamp studies in male mice, was associated with a significant rise in LH secretion. This effect of insulin is likely mediated at the hypothalamic level, because it was also found to stimulate the secretion and the expression of GnRH by hypothalamic neurons in culture. Leptin was found to potentiate the effect of insulin on GnRH secretion in vitro but was devoid of any effect on its own. These data represent the first evidence of direct insulin sensing by hypothalamic neurons involved in activating the neuroendocrine gonadotrope axis. They also demonstrate that these neurons can integrate different hormonal signals to modulate net hypothalamic GnRH output. We propose that such integration is an essential mechanism for the adaptation of reproductive function to changes in the metabolic status of an individual.

A Rural Community in a Brazilian Western Amazonian Region: Some Demographic and Epidemiological Patterns

Some demographic and epidemiological patterns of the rural population of Monte Negro, locality situated in the State of Rondônia (Brazil), Western Amazonia, are described based on a sample of 924 randomly selected individuals, approximately 10% of the whole population. The main features of this sample are (1) the illiteracy rates in the parental generation were 23% for fathers and 20% for mothers. Among children, this figure dropped to 6%; (2) housing in Monte Negro is characterized by being constructed with wood (92%), and also a floor (75%). Nevertheless, only 32% of these houses had electric energy; (3) the mean ages for the parental generation were 41.9 for males and 36.3 for females. These values for the offspring generation were 12.2 and 10.5, respectively; (4) the sex-ratio of the offspring generation was 1.32;(5) the bioassay of kinship was estimated as .033 for this long range migrant population; (6) the prevalence of some macrophage dependent infectious disease was conspicuously high; (7) the reported number of malarial episodes among males and females was statistically different, suggesting that malaria may be, in part, a "professional" disease; (8) the prevalence of serum-positive reactions against B-hepatitis is distressing. It has a strong age dependence and reaches 74% among adult males. Conversely, signs of active infection (AgHbs) rises to 16% among children.

Evaluation of two immunoassays for the measurement of human chorionic gonadotropin in urine for anti-doping purposes.

Background: Urinary human chorionic gonadotropin (hCG) concentration is routinely measured in all anti-doping laboratories to exclude the misuse of recombinant or urinary hCG preparations. In this study, extended validation of two commercial immunoassays for hCG measurements in urine was performed. Both tests were initially designed for hCG determination in human serum/plasma. Methods: Access (R) and Elecsys (R) 1010 are two automated immunoanalysers for central laboratories. The limits of detection and quantification, as well as intra-laboratory and inter-technique correlation, precision, and accuracy, were determined. Stability studies of hCG in urine following freezing and thawing cycles (n = 3) as well as storage conditions at room temperature, 4 degrees C and 20 degrees C, were performed. Results: Statistical evaluation of hCG concentrations in male urine samples (n = 2429) measured with the Elecsys (R) 1010 system enabled us to draw a skewed frequency histogram and establish a far outside value equal to 2.3 IU/L. This decision limit corresponds to the concentration at which a sportsman will be considered positive for hCG. Intra-assay precision for the Access (R) analyser was less than 4.0 A, whereas the inter-assay precision was closer to 4.5 % (concentrations of the official external controls contained between 5.5 and 195.0 IU/L). Intra and inter-assay precision for the Elecsys (R) 1010 analyser was slightly better. A good inter-technique correlation was obtained when measuring various urine samples (male and female). No urinary hCG loss was observed after two freeze/thaw cycles. On the other hand, time and inappropriate storage conditions, such as temperatures above 10 degrees C for more than 5 days, can deteriorate urinary hCG. Conclusions: Both analysers showed acceptable performances and are suitable for screening urine for anti-doping analyses. Each laboratory should validate and establish its own reference values because hCG concentrations measured in urine can be different from one immunoassay to another. The time delay between urine collection and analysis should be reduced as much as possible, and urine samples should be transported in optimal conditions to avoid a loss of hCG immunoreactivity.

The association of genetic markers and malaria infection in the Brazilian Western Amazonian region

Almost all individuals (182) belonging to an Amazonian riverine population (Portuchuelo, RO, Brazil) were investigated for ascertaining data on epidemiological aspects of malaria. Thirteen genetic blood polymorphisms were investigated (ABO, MNSs, Rh, Kell, and Duffy systems, haptoglobins, hemoglobins, and the enzymes glucose-6-phosphate dehydrogenase, glyoxalase, phosphoglucomutase, carbonic anhydrase, red cell acid phosphatase, and esterase D). The results indicated that the Duffy system is associated with susceptibility to malaria, as observed in other endemic areas. Moreover, suggestions also arose indicating that the EsD and Rh loci may be significantly associated with resistance to malaria. If statistical type II errors and sample stratification could be ruled out, hypotheses on the existence of a causal mechanism or an unknown closely linked locus involved in susceptibility to malaria infection may explain the present findings.

Epidermal growth factor receptors, testosterone levels and parotid gland changes in rats infected with Trypanosoma cruzi

It has been demonstrated that parotid glands of rats infected with Trypanosoma cruzi present severe histological alterations; changes include reduction in density and volume of the acini and duct systems and an increase in connective tissue. We evaluated the association between morphological changes in parotid glands, circulating testosterone levels and epidermal growth factor receptor (EGF-R) expression in experimental Chagas disease in rats. Animals at 18 days of infection (acute phase) showed a significant decrease in body weight, serum testosterone levels and EGF-R expression in the parotid gland compared with a control group. Since decreases in body weight could lead to a reduction in circulating testosterone concentration, we believe that the reduction in EGF-R expression in parotid glands of infected rats is due to alterations in testosterone levels and atrophy of parotid glands is caused by changes in EGF-R expression. Additionally, at 50 days (chronic phase) of infection parotid glands showed a normal histological aspect likely due to the normalization of the body weight. These findings suggest that the testosterone-EGF-R axis is involved in the histological changes.

Activation of GPER-1 estradiol receptor downregulates production of testosterone in isolated rat Leydig cells and adult human testis.

PURPOSE: Estradiol (E2) modulates testicular functions including steroidogenesis, but the mechanisms of E2 signaling in human testis are poorly understood. GPER-1 (GPR30), a G protein-coupled membrane receptor, mediates rapid genomic and non-genomic response to estrogens. The aim of this study was to evaluate GPER-1 expression in the testis, and its role in estradiol dependent regulation of steroidogenesis in isolated rat Leydig cells and human testis. MATERIALS AND METHODS: Isolated Leydig cells (LC) from adult rats and human testicular tissue were used in this study. Expression and localization studies of GPER-1 were performed with qRT-PCR, immunofluorescence, immunohistochemistry and Western Blot. Luteinizing Hormone (LH) -stimulated, isolated LC were incubated with estradiol, G-1 (GPER-1-selective agonist), and estrogen receptor antagonist ICI 182,780. Testosterone production was measured with radioimmunoassay. LC viability after incubation with G-1 was measured using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. RESULTS: GPER-1 mRNA is abundantly expressed in rat LC and human testis. Co-localization experiments showed high expression levels of GPER-1 protein in LC. E2-dependent activation of GPER-1 lowers testosterone production in isolated rats LCs and in human testis, with statistically and clinically significant drops in testosterone production by 20-30% as compared to estradiol-naïve LC. The exposure to G-1 does not affect viability of isolated LCs. CONCLUSIONS: Our results indicate that activation of GPER-1 lowers testosterone levels in the rat and human testis. The expression of GPER-1 in human testis, which lack ERα, makes it an exciting target for developing new agents affecting testosterone production in men.

Trial of recombinant follicle-stimulating hormone pretreatment for gnrh-induced fertility in patients with congenital hypogonadotropic hypogonadism.

CONTEXT AND OBJECTIVE: The optimal strategy for inducing fertility in men with congenital hypogonadotropic hypogonadism (CHH) is equivocal. Albeit a biologically plausible approach, pretreatment with recombinant FSH (rFSH) before GnRH/human chorionic gonadotropin administration has not been sufficiently assessed. The objective of the study was to test this method. DESIGN AND SETTING: This was a randomized, open-label treatment protocol at an academic medical center. PATIENTS AND INTERVENTIONS: GnRH-deficient men (CHH) with prepubertal testes (<4 mL), no cryptorchidism, and no prior gonadotropin therapy were randomly assigned to either 24 months of pulsatile GnRH therapy alone (inducing endogenous LH and FSH release) or 4 months of rFSH pretreatment followed by 24 months of GnRH therapy. Patients underwent serial testicular biopsies, ultrasound assessments of testicular volume, serum hormone measurements, and seminal fluid analyses. RESULTS: rFSH treatment increased inhibin B levels into the normal range (from 29 ± 9 to 107 ± 41 pg/mL, P < .05) and doubled testicular volume (from 1.1 ± 0.2 to 2.2 ± 0.3 mL, P < .005). Histological analysis showed proliferation of both Sertoli cells (SCs) and spermatogonia, a decreased SC to germ cell ratio (from 0.74 to 0.35), and SC cytoskeletal rearrangements. With pulsatile GnRH, the groups had similar hormonal responses and exhibited significant testicular growth. All men receiving rFSH pretreatment developed sperm in their ejaculate (7 of 7 vs 4 of 6 in the GnRH-only group) and showed trends toward higher maximal sperm counts. CONCLUSIONS: rFSH pretreatment followed by GnRH is successful in inducing testicular growth and fertility in men with CHH with prepubertal testes. rFSH not only appears to maximize the SC population but also induces morphologic changes, suggesting broader developmental roles.

Orexin/hypocretin (Orx/Hcrt) transmission and drug-seeking behavior: is the paraventricular nucleus of the thalamus (PVT) part of the drug seeking circuitry?

The orexin/hypocretin (Orx/Hcrt) system has long been considered to regulate a wide range of physiological processes, including feeding, energy metabolism, and arousal. More recently, concordant observations have demonstrated an important role for these peptides in the reinforcing properties of most drugs of abuse. Orx/Hcrt neurons arise in the lateral hypothalamus (LH) and project to all brain structures implicated in the regulation of arousal, stress, and reward. Although Orx/Hcrt neurons have been shown to massively project to the paraventricular nucleus of the thalamus (PVT), only recent evidence suggested that the PVT may be a key relay of Orx/Hcrt-coded reward-related communication between the LH and both the ventral and dorsal striatum. While this thalamic region was not thought to be part of the "drug addiction circuitry," an increasing amount of evidence demonstrated that the PVT-particularly PVT Orx/Hcrt transmission-was implicated in the modulation of reward function in general and several aspects of drug-directed behaviors in particular. The present review discusses recent findings that suggest that maladaptive recruitment of PVT Orx/Hcrt signaling by drugs of abuse may promote persistent compulsive drug-seeking behavior following a period of protracted abstinence and as such may represent a relevant target for understanding the long-term vulnerability to drug relapse after withdrawal.

Use of the Kala-Azar Detect® and IT-LEISH® rapid tests for the diagnosis of visceral leishmaniasis in Brazil

The performances of two rapid tests and a standard serological test for the diagnosis of visceral leishmaniasis (VL) were compared using sera from 193 patients with VL and 85 controls. The Kala-Azar Detect®, IT-LEISH® and IFI-LH® assays showed sensitivities of 88.1%, 93.3% and 88.6%, respectively, and specificities of 90.6%, 96.5% and 80%, respectively. The sensitivity values were similar for both rapid tests, but the specificity and positive predictive values of IT-LEISH® were higher than the corresponding values for IFI-LH®. Both rapid tests showed satisfactory performances and can be used in primary health care settings; however, IT-LEISH® permits the use of whole blood, making this assay more suitable for bedside diagnosis.

Astressin B, a nonselective corticotropin-releasing hormone receptor antagonist, prevents the inhibitory effect of ghrelin on luteinizing hormone pulse frequency in the ovariectomized rhesus monkey.

Administration of ghrelin, a key peptide in the regulation of energy homeostasis, has been shown to decrease LH pulse frequency while concomitantly elevating cortisol levels. Because increased endogenous CRH release in stress is associated with an inhibition of reproductive function, we have tested here whether the pulsatile LH decrease after ghrelin may reflect an activated hypothalamic-pituitary-adrenal axis and be prevented by a CRH antagonist. After a 3-h baseline LH pulse frequency monitoring, five adult ovariectomized rhesus monkeys received a 5-h saline (protocol 1) or ghrelin (100-microg bolus followed by 100 microg/h, protocol 2) infusion. In protocols 3 and 4, animals were given astressin B, a nonspecific CRH receptor antagonist (0.45 mg/kg im) 90 min before ghrelin or saline infusion. Blood samples were taken every 15 min for LH measurements, whereas cortisol and GH were measured every 45 min. Mean LH pulse frequency during the 5-h ghrelin infusion was significantly lower than in all other treatments (P < 0.05) and when compared with the baseline period (P < 0.05). Pretreatment with astressin B prevented the decrease. Ghrelin stimulated cortisol and GH secretion, whereas astressin B pretreatment prevented the cortisol, but not the GH, release. Our data indicate that CRH release mediates the inhibitory effect of ghrelin on LH pulse frequency and suggest that the inhibitory impact of an insufficient energy balance on reproductive function may in part be mediated by the hypothalamic-pituitary-adrenal axis.

Die loutrophóros im Hochzeits-und Begräbnisritual des 5. Jahrhunderts v. Chr. in Athen

ZUSAMMENFASSUNG Gegenstand dieser Arbeit sind rund hundert rotfigurige Darstellungen von den beiden als Lutrophoren bezeichneten Tongefässen, der dreihenkligen Lutrophoros-Hydria (LH) und der zweihenkligen Lutrophoros-Amphora (LA). Die Untersuchung geht von der Prämisse aus, dass Darstellungen der Lutrophoros einen gewissen Variantenreichtum des Brauchtums widerspiegeln können, sonst aber verbindlichen, in sich schlüssigen Vorstellungen entsprechen, die sich die Gemeinschaft von diesem Ritualobjekt macht. In Anlehnung an die herrschende Meinung wird zudem vorausgesetzt, dass LA und LH ausschliesslich im nuptialen und funeralen Bereich vorkommen, doch ist bisher in der archäologischen Forschung ihre Rolle im einzelnen ungeldärt. Anhand von literarischen Quellen und ikonographischem Material wird untersucht, inwiefern sich LA und LH in ihrer Verwendung und Bedeutung in bezug auf Hochzeit und Begräbnis unterscheiden und inwieweit der Begrifif loutrophóros auf sie zutrifft. KAPITEL I Nach frühen Quellentexten muss die Bezeichnung loutrophóros im Hochzeitsritual für eine Person und beim Begräbnis eines ehelos verstorbenen Mannes für ein Objekt verwendet worden sein. In späteren Zeugnissen wird ergänzend die Lutrophoros für die Hochzeit als hydria-ähnliches Gefäss bezeichnet, ebenso die Lutrophoros auf dem Grab; für letztere wird zusätzlich eine Kinderfigur mit Wassergefäss vorgeschlagen, was archäologisch bislang nicht bezeugt ist. Die Aussagen der Texte zum Begriff loutrophóros stimmen trotz verschiedener Widersprüche im Gesamten darin überein, dass die Aufstellung der Lutrophoros auf dem Grab von ehelos Verstorbenen eine Entsprechung zum Ritual für Hochzeitsleute darstellt; die rituelle Verwendung der Lutrophoros auf dem Grab muss also eine nuptiale Bedeutung haben. Die Untersuchung des Begriffs loutrá (,,Hoch- zeitsba") im nuptialen Bereich zeigt, dass die Lutrophoros in engem Zusammenhang mit der Quellnymphe Kallirrhoë, der Braut - nýmphè genannt - und dem Heiligtum der Nymphe, dem Hauptfundort der Lutrophoren, gesehen werden muss; allerdings bleibt die spezifische Bedeutung der loutrá ungewiss. Am ehesten werden sie dem Gedeihen der neu entstandenen Verbindung gedient haben, die sich in Anlehnung an Hesiods Werke und Tage als Neugründung eines oíkos oder als Weiteiterführung des väterlichen oíkos durch den Bräutigam umschreiben lässt. Für die bislang immer wieder vorgeschlagenen Deutungen der loutrá als Ritual für Fruchtbarkeit oder zur Reinigung gibt es frühestens ab dem 4. ]h. v. Chr. Hinweise. Die Untersuchung des Begriffs loutrá für die Waschung vor der Bestattung und ein Vergleich zwischen verschiedenen Bezeichnungen für Grab- spenden (choaí, loibaí, loutrá) und der aufgestellten Lutrophoros auf dem Grab führen entgegen bisheriger Auffassungen zum Schluss, dass die Lutrophoros auf dem Grab als Entsprechung zum Hochzeitsritual gesehen wurde und dass diesem Brauch die für Athen bekannte Vorstellung zugrunde liegt, die Verstorbenen zufrieden zu stellen. Die aufgestellte Lutrophoros könnte zum Ausdruck bringen, dass die Hinterbliebenen das der verstorbenen Person zustehende Recht auf Verehelichung gebilligt hätten. Kapitel II Beim ikonographischen Material, das sich hauptsächlich aus Vasenbildern zusammensetzt, stehen die dargestellten Gefässvarianten LA und LH im Zentrum. Nach dem ikonographischen Modell von C. Bérard fassen wir die dargestellten LA und LH im linguistischen Sinn als ikonische Zeichen auf, die nicht weiter zerlegbar sind; LA und LH stellen je eine ,,unité formelle minimale " dar. Die zentrale Frage lautet, ob Bedeutung und Verwendung der beiden ,,unités formelles minimales" LH und LA - trotz ihrer unterschiedlichen Form -in bezug auf die Funktion der loutrophóros bei der Hochzeit und beim Begräbnis ehelos Verstorbener übereinstimmen oder ob die unterschiedliche Form von LA und LH zum Ausdruck bringen soll, dass sie eben gerade nicht die gleiche Funktion haben. Beide Auffassungen sind in der bisherigen Forschung stark vertreten: R Wolters und seine Anhänger gehen davon aus, dass LA und LH gleichwertig sind; G. Kokula und mit ihr andere nehmen eine unterschiedliche, geschlechtsspezifische Verwendung an. In einer ersten formalen Analyse werden Darstellungen von LA und LH auf Lutrophoren, Lebetes gamikoi und Hydrien untersucht. Folgende Skizze zeigt die bekannten Kombinationen von dargestellten Lutrophoren: Aus der formalen Analyse lassen sich vier Schlüsse ziehen: - (ia.b) In der Regel entsprechen sich Bildträger und Bildelement; dieser Kombination muss eine Absicht zugrunde liegen, sie kann nicht willkürlich sein. - (2) Zwei LA mit Darstellungen einer LH (49 Abb. 4oa.b; 50 Abb. 4ia.b) sprechen dafür, dass im Verwendungsbereich der LA auch die LH vorkommen kann; im Gegensatz dazu fehlen aber Darstellungen einer LA auf einer LH, was vermuten lässt, dass im Verwendungsbereich der LH keine LA gebraucht wurden. - (3a.b) Zwei Darstellungen zeigen jede Variante für sich in zwei Exemplaren in der Funktion als Wasserbehälter: zwei LH (I7 Abb. 61b.c; 6za. b); zwei LA (12 Abb. 46. 47a.b); im Gegensatz dazu kommen aber beide Varianten gemeinsam in der Funktion als Wasserbehälter nicht vor, vermutlich hat sich der Anwendungsbereich der beiden Varianten unterschieden. (4a.b) Zwei Darstellungen, die beide Varianten gemeinsam zeigen, einmal im Miniaturformat (50 Abb. 41a.b), das andere Mai in monumentaler Grösse (I Abb. 1) - also nicht als Wasserbehälter -, lassen den Schluss zu, dass beide Varianten ein gemeinsames Merkmal haben müssen. Allein die formale Analyse zeigt, dass die von P Wolters angenommene Gleichwertigkeit nicht zutreffen kann. Es muss eine unterschiedliche Verwendung gegeben haben, wie bereits G. Kokula vermutet. In einer zweiten Analyse werden die Darstellungen nach der von P Bruneau vorgeschlagenen Methode in fünf Gruppen mit gleichem Schema aufgelisret. Hier wird die dargestellte LA oder LH im Vergleich zur Gesamtdarstellung ausgewertet, ohne die inhaltliche Klassifizierung vorauszusetzen. In einer Gruppe gesondert werden alle Lutrophorendarstellungen untersucht, die thematisch eindeutig zum funeralen Bereich gehören. Die aus der ersten Analyse gewonnene Annahme, dass sich LA und LH in der Verwendung unterscheiden, wird durch die zweite Analyse bestätigt und präzisiert. Für die LA kann keine eindeutige hochzeitliche Verwendung nachgewiesen werden; die LH ist das eigentliche Hochzeitsgefäss. Darüber hinaus zeigt die Untersuchung, dass alle Darstellungen einer LH oder LA inhaltlich dem nuptialen oder funeralen Kontext zugeordnet werden können; gemäss den Darstellungen wurden LA und LH nur bei Hochzeit oder Begräbnis verwendet. Nach dem hier vorgestellten Modell machen LA und LH als ikonische Zeichen eine doppelte Aussage. Sie verweisen einerseits auf den Inhalt, das hochzeitliche Wasser, und darin sind sie bedeutungsgleich. Andererseits beziehen sie sich auf die Adressaten: die LA kommt für Hochzeitsleute nicht in Frage, ihre Verwendung ist auf ehelos Verstorbene beschränkt. Die LH, das Ritualobjekt der Hochzeitsleute, kann vereinzelt auch für Verstorbene gebraucht werden. Diese zunächst irritierende Beobachtung stimmt mit den Texten gut überein, aus denen hervorgeht, dass im Ritual für ehelos Verstorbene eine Entsprechung zum Hochzeitsritual liegt; seine Bedeutung muss daher ebenfalls nuptial sein. Die Kombination von hochzeitlichen und funeralen Elementen ist daher nach dem hier vorgeschlagenen Modell im Funeralbereich - und zwar nur dort - möglich. Ebenfalls in Übereinstimmung mit den Texten lassen sich die beiden Darstellungen erklären, die LA und LH gemeinsam im Miniaturformat oder in monumentaler Grösse zeigen. Da nämlich die Mitgabe der Lutrophoros für ehelos Verstorbene auf die hochzeitlichen loutrá verweist, haben beide Gefässvarianten trotz der unterschiedlichen Verwendung eine gleichwertige Bedeutung, und daraus erklärt sich die gemeinsame Darstellung beider Varianten in einem Format, das die rituelle Verwendung ausschliesst. Durch die zweíte Analyse wird die Auffassung der geschlechtsspezifischen Verwendung von G. Kokula entkräftet. LA und LH unterscheiden sich zwar in der Verwendung, diese ist jedoch entgegen bisheriger Annahmen nicht symmetrisch. KAPITEL III Der Vergleich zwischen literarischer und ikonographischer Überlieferung zeigt einerseits das bekannte Problem, dass beide Bildelemente LA und LH nicht zweifelsfrei dem Begriff loutrophóros zugewiesen werden können. Andererseits aber stimmen Bilder und Texte in bezug auf die rituelle Funktion der loutrophóros bei Hochzeit und Begräbnis in einem hohen Mass überein, so dass widersprüchliche Aussagen nicht mehr so stark ins Gewicht fallen wie bislang. Die von einigen Lexikographen - entgegen der íkonographisch dokumentierten LA - vorgeschlagene Kinderstatue als loutrophóros auf dem Grab ehelos Verstorbener beruht vermutlich auf der Auffassung, dass sich das Erscheinungsbild der loutrophóros als Person beziehungsweise Gestalt im Hochzeíts- und Begräbnisritual entspreche. Bei dieser erst in späterer Zeit auftretenden Meinung wird deutlich, dass der ursprünglich adjektivischen Verwendung von loutrophóros, gemäss welcher bei Hochzeit und Begräbnis völlig verschiedene Wasserträger möglich sind, nicht mehr Rechnung getragen wird. KAPITEL IV Die aus den literarischen Quellen und der ikonographischen Überlieferung gezogenen Schlussfolgerungen können auch gegenüber weiteren archäologischen Gesichtspunkten, wie Gefässentwieklung und Fundorte, aufrecht erhalten bleiben. Damit würde das hier vertretene Modell unserer Forderung Rechnung tragen, dass die mit einem Ritualobjekt (hier LH und LA) verbundenen Vorstellungen kohärent sein müssen, aber eine gewisse Bandbreite von rituellen Ausführungsmodalitäten zulassen.