Disposition of MDMA and Metabolites in Human Sweat Following Controlled MDMA Administration

BACKGROUND: Understanding the excretion of 3,4-methylenedioxymethamphetamine (MDMA) and metabolites in sweat is vital for interpretation of sweat tests in drug treatment, criminal justice, and workplace programs. METHODS: Placebo, low (1.0 mg/kg), and high (1.6 mg/kg) doses of oral MDMA were given double-blind in random order to healthy volunteers (n = 15) with histories of MDMA use. Participants resided on the closed clinical research unit for up to 7 days after each dose. Volunteers wore PharmChek (R) sweat patches (n = 640) before, during, and after controlled dosing. Patches were analyzed by solid phase extraction and GC-MS for MDMA, methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxyamphetamine (HMA), and 4hydroxy-3-methoxymethamphetamine (HMMA). Limits of quantification (LOQ) were 2.5 ng/patch for MDMA and 5 ng/patch for HMA, HMMA, and MDA. RESULTS: MDMA was the primary analyte detected in 382 patches (59.7%), with concentrations up to 3007 ng/patch. MDA was detected in 188 patches (29.4%) at <172 ng/patch, whereas no HMMA or HMA was detected; 224 patches (35.0%) and 60 patches (9.4%) were positive for MDMA and MDA, respectively, at the 25-ng/patch threshold proposed by the Substance Abuse and Mental Health Services Administration. CONCLUSIONS: Sweat testing was shown to be an effective and reliable method for monitoring MDMA use in this controlled MDMA administration study. However, variability in sweat excretion suggests that results should be interpreted qualitatively rather than quantitatively. These data provide a scientific database for interpretation of MDMA sweat test results. (C) 2008 American Association for Clinical Chemistry

Effects of age on histological parameters of the sweat glands of Nellore cattle

The sweat glands are important in thermoregulation of cattle in a warm environment as they help dissipate heat through evaporation. Studies on gland histology are important to define its secretion potential and the capacity of perspiration and heat removal. The objective of this study was to determine, by histomorphometry, glandular epithelium height, the depth of the gland, length of the glandular portion and number of glands per cm2 of the sweat glands of the three age groups of Nellore cattle. Thirty females were used in this study. They were equally divided into calves, heifers and cows. Histological sections were obtained and analyzed by digital images in Trinocular BX40 Olympus microscope coupled to an Oly - 200 camera, connected to a computer. The images were obtained with microscope with 2x, 4x, 10x and 40x magnification objectives. The measurements were performed using HL Image 97 program. The height of glandular epithelium, depth of the glands, length and density of the glandular portion per cm2 , were all analyzed. The calves showed greater height of the glandular epithelium than heifers (P = 0.0024), and cows (P = 0.0191). The depth of the gland was not influenced by age. Cows had higher length of secretory portion than heifers (P = 0.0379) and calves (P = 0.0077). Heifers had a greater number of sweat glands per cm2 of skin than cows (P = 0.023). In cattle, the height of glandular epithelium and the density decreases as animals get older. On the other hand, the length of the secretor portion increases but with no changes in the depth of the sweat glands

Cystic fibrosis with normal sweat chloride concentration: case report

Cystic fibrosis is a genetic disease usually diagnosed by abnormal sweat testing. We report a case of an 18-year-old female with bronchiectasis, chronic P. aeruginosa infection, and normal sweat chloride concentrations who experienced rapid decrease of lung function and clinical deterioration despite treatment. Given the high suspicion ofcystic fibrosis, broad genotyping testing was performed, showing a compound heterozygous with deltaF508 and 3849+10kb C->T mutations, therefore confirming cystic fibrosis diagnosis. Although the sweat chloride test remains the gold standard for the diagnosis of cystic fibrosis, alternative diagnostic tests such as genotyping and electrophysiologic measurements must be performed if there is suspicion of cystic fibrosis, despite normal or borderline sweat chloride levels.

Social evolution in the sweat bee Halictus scabiosae

The evolution of eusociality is one of the major evolutionary transitions of life on earth. For investigating the conditions and processes that are central to the origin of such integrated social organization, it is best to study organisms in which individuals have retained some flexibility in their reproductive strategies. Halictid bees are especially well suited as model organisms, because they show huge variation in social systems, both within and between species. In this thesis, I investigated female reproductive strategies in the primitively eusocial bee Halictus scabiosae, with a focus on the role of helpers, in order to get insight into the mechanisms governing the evolution and maintenance of eusociality. This species produces two broods per year. The females from the first brood can stay in the natal nest to help raise a second brood of males and gynes that become the next-generation foundresses in spring. We first compared the morphology of females from the two broods, as well as the nutrition they receive as larvae. Then we conducted a helper- removal experiment in the field to quantify the effects of the presence of helpers on colony survival and productivity. Finally, we reconstructed pedigree relationships of individuals using microsatellite markers in order to detect who reproduces in the nest and how much individuals drift between nests. We found that first brood females had a uniformly small size and low fat reserves, which may be caused by the restricted pollen and nectar provisions on which they develop. Colony survival and productivity was increased by the presence of a single helper, but the effect was small and mostly limited to small colonies. By inferring parentage within and across colonies, we could determine that females from the first brood rarely reproduce in their natal nests. However, foundresses are frequently replaced, and foundresses and females from the first brood occasionally move to and reproduce in foreign colonies. As a result, colonies often contain offspring from unrelated individuals, and the relatedness of females to the brood they rear is low. Overall, this thesis shows that the reproductive system of H. scabiosae is highly flexible. The production of helpers in the first brood is important for colony success and productivity, but there is a high colony failure rate and part of the first brood females drift and reproduce in foreign nests. Both foundresses and helpers appear to be constrained by harsh environmental conditions or social factors limiting reproduction and independent colony founding. - L'origine des insectes sociaux est un domaine fascinant pour la recherche. Pour comprendre les mécanismes et les conditions qui sont nécessaires pour l'évolution et le maintien de la vie en société, il est judicieux d'étudier des sociétés primitives d'insectes, où toutes les femelles ont conservé la capacité de se reproduire, même si leur rôle comportemental dans la colonie est d'aider sans se reproduire. Une des familles d'abeilles, les halictes, est idéale pour cette sorte de recherche, en raison de la grande variabilité dans leur comportement social. Dans cette thèse, j'ai étudié les stratégies reproductives des femelles de Halictus scabiosae pour mieux comprendre les mécanismes qui influencent l'évolution de la vie en société. Cette espèce produit deux cohortes de couvain par année. Les femelles du premier couvain restent souvent dans leur nid natal pour aider à élever le deuxième couvain, tandis que les femelles du deuxième couvain s'accouplent et hibernent pour devenir les nouvelles fondatrices au printemps suivant. Nous avons d'abord comparé la morphologie des femelles issues des deux couvains ainsi que leur nutrition au stade de larve. Puis, dans une expérience sur le terrain, nous avons quantifié l'apport d'une ouvrière pour la survie et la productivité de la colonie. Finalement, nous avons reconstruit des pedigrees en utilisant des marqueurs génétiques, pour savoir qui se reproduit dans la colonie et combien d'individus migrent entre colonies. Les résultats montrent que les femelles du premier couvain sont uniformément plus petites et plus maigres, ce qui indique que les fondatrices réduisent les provisions de nourriture pour leur premier couvain afin de les inciter à aider dans le nid au lieu de se reproduire indépendamment. Dans l'expérience sur le terrain, la survie et la productivité de la colonie augmentaient avec la présence d'une ouvrière additionnelle, mais l'effet était petit et limité aux petites colonies. Par la reconstruction de pedigrees, nous pouvions constater que les femelles du premier couvain pondent rarement dans leurs nids natals. Les fondatrices cependant sont souvent remplacées en cours de saison, et migrent fréquemment entre nids, tandis que les femelles du premier couvain pondent parfois des oeufs dans des nids étrangers. De ce fait, les colonies contiennent souvent des descendants d'individus étrangers, et la parenté génétique entre les femelles et le deuxième couvain est basse. Cette thèse démontre que le système reproductif de H. scabiosae est très flexible. La production d'ouvrières est importante pour la survie de la colonie et sa productivité, mais le taux d'échec est élevé et une partie des femelles du premier couvain migrent et pondent dans une colonie étrangère. Autant les fondatrices que les ouvrières semblent être contraintes par des conditions environnementales ou sociales qui limitent la reproduction et les nouvelles fondations de colonie. - Die Entstehung von sozialen Lebensformen ist eines der wichtigsten Entwicklungen in der Geschichte des Lebens. Um die Bedingungen oder Prozesse zu verstehen, welche bei der Entstehung und dem Erhalt von sozialen Merkmalen wichtig sind, sollte man Lebewesen untersuchen, welche je nach Umwelteinflüßen ihr soziales Verhalten flexibel ändern können. Furchenbienen (Halictidae) gehören dazu. Diese weisen nämlich ein breites Spektrum verschiedener sozialer Organisationsformen auf, oftmals sogar innerhalb der einzelnen Arten. In meiner Doktorarbeit befasste ich mich mit den Fortpflanzungsstrategien der Weibchen der Skabiosen-Furchenbiene Halictus scabiosae. Diese Art produziert zwei Brüten pro Jahr. Die Weibchen der ersten Brut bleiben dabei meist als Arbeiterinnen in ihrem Geburtsnest, wohingegen die Weibchen der zweiten Brut nach der Paarung überwintern, um im nächsten Frühling neue Kolonien zu gründen. In einem ersten Schritt verglichen wir die beiden Brüten bezüglich der Grösse und der Fettreserven der Weibchen sowie der Pollen-Nektar-Vorräte für die Larven. Dann bestimmten wir in einem Feldexperiment, wieviel eine zusätzliche Arbeiterin zum Überleben und zur Produktiviät der Kolonie beiträgt. Schliesslich ermittelten wir durch genetische Tests die Verwandtschaftsbeziehungen zwischen den Bienen, um herauszufinden, wer in den Kolonien tatsächlich die Eier legt und ob und wieviel die Bienen zwischen verschiedenen Nestern wandern. Wir stellten fest, dass die Weibchen von der ersten Brut einheitlich kleiner sind und weniger Fettreserven besitzen. Das weist daraufhin, dass die Nestgründerin die erste Brut unterernährt, um die Wahrscheinlichkeit zu erhöhen, dass diese Weibchen als Arbeiterinnen im Nest bleiben anstatt sich unabhängig fortzupflanzen. Schon eine einzelne zusätzliche Arbeiterin verbesserte die Überlebenschancen und Produktivität der Kolonie, der Effekt war allerdings klein und auf kleine Kolonien beschränkt. Die Verwandtschaftsanalysen zeigten, dass die Arbeiterinnen nur sehr selten ein Ei in ihr Geburtsnest legen. Erstaunlicherweise wanderten die Nestgründerinnen oft zwischen verschiedenen Nestern. Einige Weibchen der ersten Brut wanderten auch in ein fremdes Nest und produzierten dort Nachkommen. Diese Doktorarbeit zeigt, dass die Fortpflanzungsstrategien der Skabiosen-Furchenbiene tatsächlich sehr flexibel sind. Die Anwesenheit von Arbeiterinnen ist wichtig für das Überleben und die Produktivität der Kolonie. Die Misserfolgsraten bleiben jedoch hoch, und ein Teil der Weibchen der ersten Brut pflanzt sich in fremden Nestern fort. Sowohl die Nestgründerinnen als auch die Weibchen der ersten Brut scheinen durch Umweltsbedingungen oder durch soziale Faktoren in der Wahl ihrer Fortpflanzungs¬strategie eingeschränkt zu sein.

Flexible social organization and high incidence of drifting in the sweat bee, Halictus scabiosae.

The very diverse social systems of sweat bees make them interesting models to study social evolution. Here we focus on the dispersal behaviour and social organization of Halictus scabiosae, a common yet poorly known species of Europe. By combining field observations and genetic data, we show that females have multiple reproductive strategies, which generates a large diversity in the social structure of nests. A detailed microsatellite analysis of 60 nests revealed that 55% of the nests contained the offspring of a single female, whereas the rest had more complex social structures, with three clear cases of multiple females reproducing in the same nest and frequent occurrence of unrelated individuals. Drifting among nests was surprisingly common, as 16% of the 122 nests in the overall sample and 44% of the nests with complex social structure contained females that had genotypes consistent with being full-sisters of females sampled in other nests of the population. Drifters originated from nests with an above-average productivity and were unrelated to their nestmates, suggesting that drifting might be a strategy to avoid competition among related females. The sex-specific comparison of genetic differentiation indicated that dispersal was male-biased, which would reinforce local resource competition among females. The pattern of genetic differentiation among populations was consistent with a dynamic process of patch colonization and extinction, as expected from the unstable, anthropogenic habitat of this species. Overall, our data show that H. scabiosae varies greatly in dispersal behaviour and social organization. The surprisingly high frequency of drifters echoes recent findings in wasps and bees, calling for further investigation of the adaptive basis of drifting in the social insects.

Evolution: plastic sociality in a sweat bee.

How and why do bees become social? A transplant experiment shows that sweat bees can adopt a solitary or social lifestyle in response to their environment.

Comparison of sweat rate during graded exercise and the local rate induced by pilocarpine

Centrally stimulated sweat rate produced by graded exercise until exhaustion was compared to the local sweat rate induced by pilocarpine, often used as a sweating index for healthy individuals. Nine young male volunteers (22 ± 4 years) were studied in temperate environment in two situations: at rest and during progressive exercise with 25 W increases every 2 min until exhaustion, on a cycle ergometer. In both situations, sweating was induced on the right forearm with 5 ml 0.5% pilocarpine hydrochloride applied by iontophoresis (1.5 mA, 5 min), with left forearm used as control. Local sweat rate was measured for 15 min at rest. During exercise, whole-body sweat rate was calculated from the body weight variation. Local sweat rate was measured from the time when heart rate reached 150 bpm until exhaustion and was collected using absorbent filter paper. Pharmacologically induced local sweat rate at rest (0.4 ± 0.2 mg cm-2 min-1) and mean exercise-induced whole-body sweat rate (0.4 ± 0.1 mg cm-2 min-1) were the same (P > 0.05) but were about five times smaller than local exercise-induced sweat rate (control = 2.1 ± 1.4; pilocarpine = 2.7 ± 1.2 mg cm-2 min-1), indicating different sudorific mechanisms. Both exercise-induced whole-body sweat rate (P < 0.05) and local sweat rate (P < 0.05) on control forearm correlated positively with pilocarpine-induced local sweat rate at rest. Assuming that exercise-induced sweating was a result of integrated physiological mechanisms, we suggest that local and whole-body sweat rate measured during graded exercise could be a better sweating index than pilocarpine.

Effect of age and gender on sweat lactate and ammonia concentrations during exercise in the heat

The dependence of sweat composition and acidity on sweating rate (SR) suggests that the lower SR in children compared to adults may be accompanied by a higher level of sweat lactate (Lac-) and ammonia (NH3) and a lower sweat pH. Four groups (15 girls, 18 boys, 8 women, 8 men) cycled in the heat (42ºC, 20% relative humidity) at 50% VO2max for two 20-min bouts with a 10-min rest before bout 1 and between bouts. Sweat was collected into plastic bags attached to the subject's lower back. During bout 1, sweat from girls and boys had higher Lac- concentrations (23.6 ± 1.2 and 21.2 ± 1.7 mM; P < 0.05) than sweat from women and men (18.2 ± 1.9 and 14.8 ± 1.6 mM, respectively), but Lac- was weakly associated with SR (P > 0.05; r = -0.27). Sweat Lac- concentration dropped during exercise bout 2, reaching similar levels among all groups (overall mean = 13.7 ± 0.4 mM). Children had a higher sweat NH3 than adults during bout 1 (girls = 4.2 ± 0.4, boys = 4.6 ± 0.6, women = 2.7 ± 0.2, and men = 3.0 ± 0.2 mM; P < 0.05). This difference persisted through bout 2 only in females. On average, children's sweat pH was lower than that of adults (mean ± SEM, girls = 5.4 ± 0.2, boys = 5.0 ± 0.1, women = 6.2 ± 0.5, and men = 6.2 ± 0.4 for bout 1, and girls = 5.4 ± 0.2, boys = 6.5 ± 0.5, women = 5.2 ± 0.2, and men = 6.9 ± 0.4 for bout 2). This may have favored NH3 transport from plasma to sweat as accounted for by a significant correlation between sweat NH3 and H+ (r = 0.56). Blood pH increased from rest (mean ± SEM; 7.3 ± 0.02) to the end of exercise (7.4 ± 0.01) without differences among groups. These results, however, are representative of sweat induced by moderate exercise in the absence of acidosis.

Intraspecific and interspecific social variation in the sweat bee Lasioglossum Malachurum and other members of the subgenus Evylaeus

Sweat bees exhibit a range of social behaviours, from solitary nesting, in which no workers are produced, to strong eusociality, in which workers exhibit a high degree of altruism, behaviour that is measured by the degree of personal reproductive sacrifice. Field studies were carried out for seven weeks during May-June 2000 in southern Greece in order to investigate intraspecific social variation, and test the hypothesis of a north-south cline of decreasing eusociality in the obligately eusocial sweat bee L. (E.) malachurum. A comparative study, using principal components analysis, was performed to determine if patterns of intraspecific social variation in L. malachurum reflect the patterns of social variation within the subgenus, Evylaeus, as a whole. The results of the field study reveal that, in Greece, two worker broods were produced followed by a third brood consisting of gynes, males and some workers, indicating that there was an overlap in worker and gyne production. There was strong caste distinction between queens and workers. Workers actively foraged and participated in nest construction as most workers (58%, n=303) had a high degree of mandibular wear. Workers did not participate in the oviposition of Brood 3 gynes since only 0.7% (n=278) of workers were mated. Furthermore, queen survival until the end of Brood 3 and a substantial size differential of 10.6% between queens and workers suggested that queen domination over worker behaviour during the early to mid-part of the colony cycle was plausible. Male production in Brood 3 by some workers was likely, since the timing of worker ovarian development corresponded with the timing of male production. These findings suggest that workers of the first two broods were primarily altruistic, but some (28%) Brood 1 (9%) and Brood 2 (19%) workers produced males, indicating that the degree of altruistic behaviour declined during the lifetime of the colony. In comparison with other L. malachurum populations in Europe, the Greek population of L. malachurum had a weaker social level as a result of the higher proportion of workers potentially involved in male production, thus 3 supporting the hypothesis of a southerly cline of decreasing eusociality. Furthermore, intraspecific variation in social level across Europe appears to be due to longer breeding seasons in more southerly locations that would promote the production of larger colonies and provide opportunities for workers to evade queen control. The comparative study using principal components analysis on 20 solitary (of the subgenera Evylaeus and Lasioglossum), eusocial and socially polymorphic Evylaeus species and populations reveals that six traits are closely associated with stronger eusociality in Evylaeus. These traits are: (1) a reduction in the proportion of males in the early brood(s); (2) a reduction in the proportion of females that mate; (3) an increase in the mean number of first brood workers; (4) a reduction in the proportion of females with developed ovaries; (5) an increase in size dimorphism between castes, and (6) nest guarding. These are traits that most significantly define principal component one and therefore distinguish social type as indicated by a clear separation of the eusocial and the solitary populations, with a socially polymorphic species falling in between. Furthermore, most of these traits are under foundress control and may suggest that the evolutionary loss or gain of eusociality is based on selection pressures on a founding female. Colony size and female ovarian development are common factors distinguishing social variation in L. malachurum and within the subgenus as a whole. The principal components analysis excluding the solitary species and the socially aberrant L. marginatum populations show the L. malachurum populations separated based on an increasing proportion of workers with developed ovaries as populations are found more south, lending further support to the hypothesis of a north-south cline of decreasing eusociality.