Dance clubbing on MDMA and during abstinence from Ecstasy/MDMA: prospective neuroendocrine and psychobiological changes.

BACKGROUND/AIMS: The present study is the first to prospectively compare a group of recreational Ecstasy users when dance clubbing on 3,4-methylenedioxymethamphetamine (MDMA) and when clubbing during abstinence from Ecstasy/MDMA. METHODS: Twelve normal healthy volunteers (mean age = 23.2 years) were assessed at a Saturday night dance club under self-administered MDMA. On the other weekend they went to the same dance club without taking MDMA (order counterbalanced). Both conditions involved 5 test sessions conducted at similar times: pre-drug baseline, 1 h post-drug clubbing, 2.5 h post-drug clubbing, and 2 and 4 days later. The assessments included body and ambient temperature, physical activity (pedometer), as well as self-ratings for mood state, physical activity, thermal comfort and thirst. Saliva samples were analyzed for MDMA, cortisol and testosterone. RESULTS: The cortisol levels increased significantly by 800% when dance clubbing on MDMA, while testosterone increased significantly by 75%; neither neuroendocrine measure was altered during abstinence. Saliva analyses confirmed the presence of MDMA when dancing on Ecstasy and its absence when dancing off Ecstasy. The pedometer values and self-rated levels of dancing were similar at both weekends. Hot and cold flushes and feeling hot increased significantly under MDMA. The mean body temperature did not change significantly, although there was a borderline trend for increased values after MDMA. Feelings of happiness and excitement increased under MDMA, although they were not significantly greater than when clubbing during abstinence. CONCLUSIONS: Neurohormonal release may be an important part of the acute MDMA experience. The large cortisol increase provides further data on the bioenergetic stress model of recreational Ecstasy/MDMA.

Extracellular loops and ligand binding to a subfamily of Family A G-protein-coupled receptors.

GPCRs (G-protein-coupled receptors) are a large family of structurally related proteins which mediate their effects by coupling to G-proteins. The V(1a)R (V(1a) vasopressin receptor) is a member of a family of related GPCRs that are activated by vasopressin {AVP ([Arg(8)]vasopressin)}, OT (oxytocin) and related peptides. These receptors are members of a subfamily of Family A GPCRs called the neurohypophysial peptide hormone receptor family. GPCRs exhibit a conserved tertiary structure comprising a bundle of seven TM (transmembrane) helices linked by alternating ECLs (extracellular loops) and ICLs (intracellular loops). The cluster of TM helices is functionally important for ligand binding, and, furthermore, activation of GPCRs involves movement of these TM helices. Consequently, it might be assumed that the extracellular face of GPCRs is composed of peptide linkers that merely connect important TM helices. However, using a systematic mutagenesis approach and focusing on the N-terminus and the second ECL of the V(1a)R, we have established that these extracellular domains fulfil a range of important roles with respect to GPCR signalling, including agonist binding, ligand selectivity and receptor activation.

Ligand binding and activation of the CGRP receptor.

The receptor for CGRP (calcitonin gene-related peptide) is a heterodimer between a GPCR (G-protein-coupled receptor), CLR (calcitonin receptor-like receptor) and an accessory protein, RAMP1 (receptor activity-modifying protein 1). Models have been produced of RAMP1 and CLR. It is likely that the C-terminus of CGRP interacts with the extracellular N-termini of CLR and RAMP1; the extreme N-terminus of CLR is particularly important and may interact directly with CGRP and also with RAMP1. The N-terminus of CGRP interacts with the TM (transmembrane) portion of the receptor; the second ECL (extracellular loop) is especially important. Receptor activation is likely to involve the relative movements of TMs 3 and 6 to create a G-protein-binding pocket, as in Family A GPCRs. Pro(321) in TM6 appears to act as a pivot. At the base of TMs 2 and 3, Arg(151), His(155) and Glu(211) may form a loose equivalent of the Family A DRY (Asp-Arg-Tyr) motif. Although the details of this proposed activation mechanism clearly do not apply to all Family B GPCRs, the broad outlines may be conserved.

Heterodimers and family-B GPCRs: RAMPs, CGRP and adrenomedullin.

RAMPs (receptor activity-modifying proteins) are single-pass transmembrane proteins that associate with certain family-B GPCRs (G-protein-coupled receptors). Specifically for the CT (calcitonin) receptor-like receptor and the CT receptor, this results in profound changes in ligand binding and receptor pharmacology, allowing the generation of six distinct receptors with preferences for CGRP (CT gene-related peptide), adrenomedullin, amylin and CT. There are three RAMPs: RAMP1-RAMP3. The N-terminus appears to be the main determinant of receptor pharmacology, whereas the transmembrane domain contributes to association of the RAMP with the GPCR. The N-terminus of all members of the RAMP family probably contains two disulphide bonds; a potential third disulphide is found in RAMP1 and RAMP3. The N-terminus appears to be in close proximity to the ligand and plays a key role in its binding, either directly or indirectly. BIBN4096BS, a CGRP antagonist, targets RAMP1 and this gives the compound very high selectivity for the human CGRP(1) receptor.

The pharmacology of CGRP-responsive receptors in cultured and transfected cells.

Historically, CGRP receptors have been classified as CGRP(1) or CGRP(2) subtypes, chiefly depending on their affinity for the antagonist CGRP(8-37). It has been shown that the complex between calcitonin receptor-like receptor (CRLR or CL) and receptor activity modifying protein (RAMP) 1 provides a molecular correlate for the CGRP(1) receptor; however, this does not explain the range of affinities seen for CGRP(8-37) in isolated tissues. It is suggested that these may largely be explained by a combination of methodological factors and CGRP-responsive receptors generated by CL and RAMP2 or RAMP3 and complexes of RAMPs with the calcitonin receptor.

CL/RAMP2 and CL/RAMP3 produce pharmacologically distinct adrenomedullin receptors: a comparison of effects of adrenomedullin22-52, CGRP8-37 and BIBN4096BS.

Adrenomedullin (AM) has two known receptors formed by the calcitonin receptor-like receptor (CL) and receptor activity-modifying protein (RAMP) 2 or 3: we report the effects of the antagonist fragments of human AM and CGRP (AM22-52 and CGRP8-37) in inhibiting AM at human (h), rat (r) and mixed species CL/RAMP2 and CL/RAMP3 receptors transiently expressed in Cos 7 cells or endogenously expressed as rCL/rRAMP2 complexes by Rat 2 and L6 cells. AM22-52 (10 microM) antagonised AM at all CL/RAMP2 complexes (apparent pA2 values: 7.34+/-0.14 (hCL/hRAMP2), 7.28+/-0.06 (Rat 2), 7.00+/-0.05 (L6), 6.25+/-0.17 (rCL/hRAMP2)). CGRP8-37 (10 microM) resembled AM22-52 except on the rCL/hRAMP2 complex, where it did not antagonise AM (apparent pA2 values: 7.04+/-0.13 (hCL/hRAMP2), 6.72+/-0.06 (Rat2), 7.03+/-0.12 (L6)). On CL/RAMP3 receptors, 10 microM CGRP8-37 was an effective antagonist at all combinations (apparent pA2 values: 6.96+/-0.08 (hCL/hRAMP3), 6.18+/-0.18 (rCL/rRAMP3), 6.48+/-0.20 (rCL/hRAMP3)). However, 10 microM AM22-52 only antagonised AM at the hCL/hRAMP3 receptor (apparent pA2 6.73+/-0.14). BIBN4096BS (10 microM) did not antagonise AM at any of the receptors. Where investigated (all-rat and rat/human combinations), the agonist potency order on the CL/RAMP3 receptor was AM approximately betaCGRP>alphaCGRP. rRAMP3 showed three apparent polymorphisms, none of which altered its coding sequence. This study shows that on CL/RAMP complexes, AM22-52 has significant selectivity for the CL/RAMP2 combination over the CL/RAMP3 combination. On the mixed species receptor, CGRP8-37 showed the opposite selectivity. Thus, depending on the species, it is possible to discriminate pharmacologically between CL/RAMP2 and CL/RAMP3 AM receptors.

Interaction of calcitonin-gene-related peptide with its receptors.

The receptor for calcitonin-gene-related peptide (CGRP) is a heterodimer formed by calcitonin-receptor-like receptor (CRLR), a type II (family B) G-protein-coupled receptor, and receptor-activity-modifying protein 1 (RAMP1), a single-membrane-pass protein. It is likely that the first seven or so amino acids of CGRP (which form a disulphide-bonded loop) interact with the transmembrane domain of CRLR to cause receptor activation. The rest of the CGRP molecule falls into three domains. Residues 28-37 and 8-18 are normally required for high-affinity binding, while residues 19-27 form a hinge region. The 28-37 region is almost certainly in direct contact with the receptor; 8-18 may make additional receptor contacts or may stabilize an appropriate conformation of 28-37. It is likely that these regions of CGRP interact both with CRLR and with the extracellular domain of RAMP1.

Clay pica has no hematologic or metabolic correlate in chronic hemodialysis patients.

OBJECTIVE: Clay pica is a form of compulsive ingestion of non-nutritive substances frequently practiced by dialysis patients. Its consequences are unknown. In this study, we evaluated the effect of regular consumption of clay on hematologic and metabolic profiles in hemodialysis patients. DESIGN: A prospective, case-control study with use of structured questionnaire. SETTING: Free-standing hemodialysis units. PATIENTS: One hundred thirty-eight patients on hemodialysis for at least 12 months were interviewed. Thirteen of 138 (9.4%) confessed to clay pica. Ten randomly selected patients with no history of pica served as control. INTERVENTION: Average of all laboratory profiles and interdialytic weight gain (IDWG) over a 3-month period were recorded. Assay of the aluminum (Al), silica (Si), and iron (Fe) content of commercially purchased clay was performed. MAIN OUTCOME MEASURE: Comparison of laboratory profiles and IDWG between cases and control. Estimation of the daily consumption of Al, Fe, and Si from clay and their relationship to the laboratory profiles. RESULTS: There was no statistically significant difference in the levels of Al, albumin, calcium, ferritin, hematocrit, iron saturation, phosphorus, and IDWG between pica cases and control. Iron was significantly higher in pica patients (13.0 +/- 7.9 micromol/L v 7.5 +/- 2.5 micromol/L, P =.04), but potassium was higher among control than pica cases (4.9 +/- 0.7 mmol/L v 4.4 +/- 0.6 mmol/L, P =.07). Estimated metal exposure from daily clay consumption per patient were: Al (1-2 mg), Fe (11-23.5 mg), and Si (2-4.5 g). Multivariate logistic regression analysis failed to show any association between clay consumption and nutrition, anemia, or mineral metabolism (R(2) = 0.0, P =.79). CONCLUSION: Clay pica does not appear to be detrimental to the hematologic and metabolic milieu of hemodialysis patients. The practice should, however, be discouraged, because of potential for ingestion of unknown substances, and reported potential for gastrointestinal complications.

Tunneled hemodialysis catheter survival: comparison of radiologic and surgical implantation.

Cuffed, tunneled hemodialysis catheters (caths) are often implanted in the operating rooms (OR) by surgeons or by interventional radiologists in radiology suites (RS). Comparative outcome studies between OR and RS placed caths are few and tend to favor the specialty of the authors. In this longitudinal study, we monitored cath survival in patients while awaiting maturation of their fistulae, and compared outcomes between OR and RS placement. A total of 95 caths were placed in 50 patients between July 1996 and July 1999. Radiologically placed caths had a shorter primary patency duration than OR placed caths (80 +/- 40 days vs. 100 +/- 31 days, p = 0.04) and a lower primary patency rate at 120 days than OR placed caths (42% vs. 67%, p = 0.04). Cumulative infection rate per 1,000 catheter days was higher in RS than OR cases (3.8 Vs 2.2, p = 0.09), whereas mean sepsis free duration was shorter in RS than OR (60 +/- 45 days vs. 88 +/- 40 days, p = 0.02). The risk of infection was 1.7 times greater in RS than OR cases (chi-square = 6.4, p = 0.01). The RS placed caths also had a higher rate of primary nonfunction (31% vs. 8.3%, p = 0.04) and bleeding complications (42% vs. 17%, p = 0.04), but significantly shorter procedure scheduling time than OR cases (1.1 +/- 0.3 days vs. 2.5 +/- 0.6 days, p < 0.0001). In conclusion, radiologically placed caths seem to have higher rates of infection, bleeding, and functional failure but shorter scheduling time than surgically placed caths. Discussions are under way to improve the survival of RS placed caths at our affiliated hospitals.

Merkel cell carcinoma.

Merkel cell carcinoma is an unusual primary cutaneous tumor with an aggressive biologic nature. Following surgical treatment, 40% of patients have local recurrences develop, 55% have regional lymph node metastases develop, and 49% have distant metastases develop. We have treated four patients with Merkel cell carcinoma; only one of the four patients was alive and well after 2 years. Two patients died of metastatic disease, one at 11 months following initial treatment and one at 39 months. The fourth patient had a rapid recurrence following initial treatment and is currently in remission following chemotherapy for regional metastases. Recent reports indicate that chemotherapy may be helpful in treating patients with recurrent or metastatic Merkel cell carcinoma.

Blast tattoos resulting from black powder firearms.

Blast tattoos occur when fragments of gunpowder are propelled into the skin during a firearm discharge. This unusual type of injury is often associated with accidents involving replica firearms. Some of the more superficially embedded powder fragments may be removed with dermabrasion; however, deeper fragments may cause permanent tattooing. Blast injuries to the face also have the potential of causing permanent eye injury.

Treatment of multiple facial neurofibromas with dermabrasion.

Three patients with multiple facial neurofibromas were treated by dermabrasion with good cosmetic results. Wound healing was entirely normal and no complications were observed. There was no evidence of accelerated regrowth of tumors during follow-up.