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An assay to detect UO(2)(2+) complexation was developed based on the chrome azurol S (CAS) assay for siderophores (B. Schwyn and J. B. Neilands, Anal. Biochem. 160:47-56, 1987) and was used to investigate the ability of fungal metabolites to complex actinides. In this assay the discoloration of two dyed agars (one containing a CAS-Fe(3+) dye and the other containing a CAS-UO(2)(2+) dye) caused by ligands was quantified. The assay was tested by using the siderophore desferrioxamine B (DFO), and the results showed that there was a regular, reproducible relationship between discoloration and the amount of siderophore added. The ratio of the discoloration on the CAS-UO(2)(2+) agar to the discoloration on the CAS-Fe(3+) agar was independent of the amount of siderophore added. A total of 113 fungi and yeasts were isolated from three soil samples taken from the Peak District National Park. The fungi were screened for the production of UO(2)(2+) chelators by using the CAS-based assay and were also tested specifically for hydroxamate siderophore production by using the hydroxamate siderophore auxotroph Aureobacterium flavescens JG-9. This organism is highly sensitive to the presence of hydroxamate siderophores. However, the CAS-based assay was found to be less sensitive than the A. flavescens JG-9 assay. No significant difference between the results for each site for the two tests was found. Three isolates were selected for further study and were identified as two Pencillium species and a Mucor species. Our results show that the new assay can be effectively used to screen fungi for the production of UO(2)(2+) chelating ligands. We suggest that hydroxamate siderophores can be produced by mucoraceous fungi.

Pharmacologically active preparations directed towards modulating muscarinic receptor activity in the eye have been used for over 2000 years when extracts from Atropa belladonna were first applied to enhance eye appearance through pupillary dilation. The first clinically active drugs targeting a specific eye disease were anticholinesterases (e.g. ecothiophate) applied as eye drops to treat glaucoma in the 1960's. However, cataract was soon detected as a relatively frequent side effect and such drugs are now only used to treat glaucoma as a last resort. As muscarinic agonists have been found to reduce intraocular pressure both by decreasing aqueous humour production (through Na,K-ATPase pump inhibition) and increasing outflow (by muscle contraction), it is likely that treatments will be developed that target specific muscarinic subtypes. Recently, it has been shown that the M1 receptor subtype predominates in the lens. It is therefore important that this subtype is not targeted in future ocular therapies so that the side-effect of cataract is avoided. Form-deprived myopia resulting from an increased axial length in the affected eye can be reduced by the application of atropine. This effect has been achieved both in a chick model system and in human clinical trials, and in the former system atropine has been shown to reduce the production of scleral extracellular proteins. Carbachol stimulates tear fluid production through the activation of muscarinic receptors. Interestingly, at least part of the stimulation occurs via epidermal growth factor (EGF) receptors and although the precise signalling mechanisms are not completely understood, it has been shown that calcium mobilisation plays a critical role in both muscarinic and EGF receptor activity. It should be noted that in the four examples described above, the cell types responsible for producing the physiological output are non-neuronal in origin. Therefore cholinergic receptor activation plays diverse roles in the eye and pharmacological intervention based on specific receptor sub-types has potential benefit in a number of ocular problems. However, potential side effects have also recently been identified., (Copyright 2003 Elsevier Science Inc.)

The solution chemistry of uranyl ion with iminodiacetate (IDA) and oxydiacetate (ODA) was investigated using NMR and EXAFS spectroscopies, potentiometry, and calorimetry. From the NMR and EXAFS data and depending on stoichiometry and pH, three types of metal:ligand complex were identified in solution in the pH range 3-7: 1:1 and 1:2 monomers; a 2:2 dimer. From NMR and EXAFS data for the IDA system and previous studies, we propose the three complex types are [UO(2)(IDA)(H(2)O)(2)], [UO(2)(IDA)(2)](2)(-), and [(UO(2))(2)(IDA)(2)(mu-OH)(2)](2)(-). From EXAFS spectroscopy, similar 1:1, 2:2, and 1:2 complexes are found for the ODA system, although (13)C NMR spectroscopy was not a useful probe in this system. For the 1:1 and 1:2 complexes in solution, EXAFS spectroscopy is ambiguous because the data can be fitted with either a long U-N/O(ether) value (ca. 2.9 A) suggesting 1,7-coordination of the ligand or a U-C interaction at a similar distance, consistent with terminal bidentate coordination. However, the NMR data of the IDA system suggest that 1,7-coordination is the more likely. The stability constants of the three complexes were determined by potentiometric titrations; the log beta values are 9.90 +/-, 16.42 +/-, and 10.80 +/- for the 1:1, 1:2, and 2:2 uranyl-IDA complexes, respectively, and 5.77 +/-, 7.84 +/-, and 4.29 +/- for the 1:1, 1:2, and 2:2 uranyl-ODA complexes, respectively. The thermodynamic constants for the complexes were calculated from calorimetric titrations; the enthalpy changes (kJ mol(-)(1)) and entropy changes (J K(-)(1) mol(-)(1)) of complexation for the 1:1, 1:2, and 2:2 complexes respectively are the following. IDA: 12 +/- 2, 230 +/- 8; 8 +/- 2, 151 +/- 9; -33 +/- 3, -283 +/- 11. ODA: 26 +/- 2, 198 +/- 12; 20 +/- 2, 106 +/- 8; -24 +/- 2; -219 +/- 8.

The dodecametallic Cr(III) cluster has an S = 6 ground state with an axial zero-field splitting (ZFS) of DS=6 = +0.088 cm-1. Analysis of high-resolution optical data (MCD) allows us to determine the single-ion ZFS of the constituent Cr(III) ions directly (D = -1.035 cm-1). A vector coupling analysis demonstrates that the cluster ZFS is almost entirely single-ion in origin. Thus, the relative orientations of the local and cluster magnetic axes can lead to cluster ZFS of opposite sign to the single-ion even when this is the only significant contribution.

The reaction of suitably disubstituted alkynes with diisopropyl xanthogen disulfide gives differentially substituted 4,5-disubstituted-1,3-dithiol-2-ones as proligands for metal complexes related to the molybdenum cofactor.

The reaction of MnF3 with benzotriazole (BTAH) in hot MeOH gives the Mn(III) complex [Mn26O16(OH)10(OMe)6(F)10(BTA)20(BTAH)2(MeOH)13(H2O)] which displays hysteresis and temperature-independent magnetisation relaxation at low temperature.

Treatment of [UO2Cl2(thf)3] in thf with one equivalent of [Na(CH(Ph2P = NSiMe3)2)] yields an unusual uranyl chloro-bridged dimer containing a uranium(VI)-carbon bond as part of a tridentate bis(iminophosphorano)methanide chelate complex. The methine carbon is displaced significantly from the uranyl equatorial plane.

Multifrequency continuous wave EPR spectra (4-34 GHz) on a powder of the title compound are consistent with a spin-triplet state. This arises from interaction between centrosymmetrically related pairs of copper(II) ions in the solid. The spectra at all frequencies have been simulated with a single set of spin-Hamiltonian parameters. The results show that there is noncoincidence between the principal axes of the g-matrices on each copper center and those of the zero-field splitting (D) tensor. This noncoincidence is a single rotation of 33 degrees +/- 2 degrees. The parameters from the powder spectra have been verified by a subsequent single-crystal EPR study which yielded the spin-Hamiltonian parameters g(XX) = 2.074, g(YY) = 2.093, g(ZZ) = 2.385, D(XX) = +/-0.0228 cm(-1), D(YY) = +/-0.0211 cm(-1), D(ZZ) = -/+0.0439 cm(-1) with Euler angles of alpha = 179 degrees, chi = 33.4 degrees, and gamma = 328 degrees. Analysis of the zero-field splitting tensor in terms of exchange indicates that the interaction between the pairs of copper(II) ions is almost entirely dipolar in origin. This study shows that multifrequency EPR spectroscopy on powders, coupled with spectrum simulation, can detect and measure noncoincidence between the principal axes of the g-matrix and zero-field splitting tensor, and does not necessarily require the presence of metal hyperfine interactions.

Eight uranyl compounds containing the dicarboxylate ligands iminodiacetate (IDA) or oxydiacetate (ODA) have been characterized in the solid state. The published polymeric structures for [UO(2)(C(4)H(6)NO(4))(2)] and [UO(2)(C(4)H(4)O(5))](n) have been confirmed, while Ba[UO(2)(C(4)H(5)NO(4))(2)] x 3H(2)O, [(CH(3))(2)NH(CH(2))(2)NH(CH(3))(2)][UO(2)(C(4)H(4)O(5))(2)] [orthorhombic space group Pnma, a = 10.996(5) A, b = 21.42(1) A, c = 8.700(3) A, Z = 4], and [C(2)H(5)NH(2)(CH(2))(2)NH(2)C(2)H(5)][UO(2)(C(4)H(4)O(5))(2)] [monoclinic space group P2(1)/n, a = 6.857(3) A, b = 9.209(5) A, c = 16.410(7) A, beta = 91.69(3), Z = 2] contain monomeric anions. The distance from the uranium atom to the central heteroatom (O or N) in the ligand varies. Crystallographic study shows that U-heteroatom (O/N) distances fall into two groups, one 2.6-2.7 A in length and one 3.1-3.2 A, the latter implying no bonding interaction. By contrast, EXAFS analysis of bulk samples suggests that either a long U-heteroatom (O/N) distance (2.9 A) or a range of distances may be present. Three possible structural types, two symmetric and one asymmetric, are identified on the basis of these results and on solid-state (13)C NMR spectroscopy. The two ligands in the complex can be 1,4,7-tridentate, giving five-membered rings, or 1,7-bidentate, to form an eight-membered ring. (C(4)H(12)N(2))[(UO(2))(2)(C(4)H(5)NO(4))(2)(OH)(2)] x 8H(2)O [monoclinic space group P2(1)/a, a = 7.955(9) A, b = 24.050(8) A, c = 8.223(6) A, beta = 112.24(6), Z = 2], (C(2)H(10)N(2))[(UO(2))(2)(C(4)H(5)NO(4))(2)(OH)(2)] x 4H(2)O, and (C(6)H(13)N(4))(2)[(UO(2))(2)(C(4)H(4)O(5))(2)(OH)(2)] x 2H(2)O [monoclinic space group C2/m, a = 19.024(9) A, b = 7.462(4) A, c = 2.467(6) A, beta = 107.75(4), Z = 4] have a dimeric structure with two capping tridentate ligands and two mu(2)-hydroxo bridges, giving edge-sharing pentagonal bipyramids.

A new N,O-bidentate pro-ligand (HL), [ML2] (M = Cu, Zn) and [CuL2][BF4] have been synthesised; [CuL2].4DMF and [CuL2][BF4].2CH2Cl2 have been crystallographically and spectroscopically characterised; these data indicate that [CuL2]+ cations are constituted as [Cu2+(L.)(L-)]+ and involve the phenoxyl radical L..

Purpose: To investigate regional differences in Ca(2+) mobilization kinetics in the intact human lens produced by exposure to agonists of tyrosine-kinase and G-protein-coupled receptors and to characterize the major receptor subtypes involved in Ca(2+) signaling in the different regions., Methods: Whole human lenses were placed anterior side down in a plastic chamber and perifused with artificial aqueous humor (AAH) at 30 degrees C. After fura-2 incorporation, cytosolic Ca(2+) levels were monitored by using epifluorescence techniques in either the equatorial or central anterior epithelial cells of the intact lens. Agonists dissolved in AAH were applied to the lens in successive short pulses., Results: Central anterior lens epithelial cells produced a large response to 10 microM acetylcholine (ACh) and histamine; only a small response to adenosine triphosphate (ATP); and no response to 10 microM adrenalin, 10 ng/ml epithelial growth factor (EGF) or TGF alpha, or 50 ng/ml platelet-derived growth factor (PDGF)-AB. Conversely, the equatorial cells produced a strong response to 10 microM ATP and histamine, 10 ng/ml EGF (or TGF alpha), and 50 ng/ml PDGF-AB, but failed to respond to 10 microM ACh or 10 microM adrenalin. The EGF-induced response in the equatorial cells was blocked completely by tyrphostin (AG1478), a specific inhibitor of the EGF receptor tyrosine kinase. Carbachol, a nonhydrolyzable analogue of ACh, and pilocarpine, the M1 muscarinic receptor-specific agonist, both produced the same trend of response amplitude elicited by ACh in each region of the lens. The potency order of purinergic agonist-induced Ca(2+) mobilization at the equator was consistent with the P2Y(2) receptor subtype. The histamine-induced response was abolished by 10 microM triprolidine, a specific H(1) receptor antagonist, but remained unaffected by the specific H(2) and H(3) antagonists, ranitidine and thioperamide, respectively. CONCLUSIONS. There is a spatial heterogeneity in functional receptor activity in different regions of the whole lens. The important growth factor receptors for EGF and PDGF are functionally active only in the equatorial cells of the mature human lens. This study further shows that the ACh, histamine, and ATP-induced responses arise from the activation of M1 muscarinic, H(1) histamine, and P2Y(2) purinergic receptors, respectively.

Purpose: Localized cortical cataracts in the human lens have been shown to involve a selective increase in calcium with no change in sodium content. Recent studies in the rat lens in vitro have shown that the store-operated channel is highly selective for calcium over sodium, and therefore this channel was characterized further in human lens cells., Methods: Human primary cultures were initiated from epithelial explants and passaged onto coverslips. After incorporating Fura-2, agonist- or thapsigargin-induced changes in cytosolic calcium were monitored and calibrated using fluorometric digital imaging techniques., Results: Histamine and adenosine triphosphate (ATP; 10 microM) induced a large transient increase in cytosolic calcium followed by a maintained lower plateau phase in the continued presence of the calcium-signaling agonist. The second phase was abolished by removing external calcium and represented the contribution from the store-operated influx. The store-operated pathway was blocked by inorganic agents such as zinc and nickel (100 microM) but was insensitive to the voltage-sensitive calcium channel blocker, nifedipine (1 mM). Depolarizing the membrane voltage by raising the external potassium (75 mM) also blocked the influx. Similar results were obtained if the store was first emptied directly using thapsigargin (1 microM), and with this agent it was also possible to observe the very slow activation and inactivation kinetics (>10 seconds) of the channel. Addition of manganese to the bathing medium initiated a quench of Fura-2 isobestic fluorescence that was enhanced 2.9 +/- 0.3-fold after 10 microM ATP addition. There was a delay of 82 +/- 16 seconds between initiation of the calcium spike and the Mn2+ quench rate, indicating the presence of a delayed entry pathway. In the resting state, removal of, or increasing extracellular calcium concentration 10-fold did not perturb the level of cytosolic Ca2+. Similar maneuvers performed after agonist- or thapsigargin-induced store depletion of intracellular stores brought about dramatic changes in cytosolic Ca2+ consistent with the activation of a Ca2+ entry pathway. Lower concentrations of agonist induced oscillations of Ca2+ that continued for a short time in Ca-free solution. No increase in Mn2+ quench rate was associated with oscillations. A 100-microM zinc- and KCl-induced blockade of Ca2+ entry had no effect on the form of agonist-induced oscillations. Inhibition of Ca2+ influx by zinc (100 microM) converted a sustained Ca2+ response to a train of repetitive Ca2+ spikes., Conclusions: Human lens cells normally have very low Ca2+ permeability. Depletion of intracellular stores by agonists or thapsigargin initiates a Ca2+ entry pathway that is not required for the Ca2+ oscillations induced by low concentrations of agonist. This potentially provides a signal transduction mechanism with minimal risk of Ca2+ overload to the lens, whereas overactivation of the store-operated channel is a possible way of increasing calcium in the lens and could explain the distribution found in localized cataracts.

The stereochemistry of isolated natural product Amavadin, which contains a 1:2 complex of V(IV) with N-hydroxyimino-2,2'-dipropionic acid (HIDPAH(3)), and some synthetic complexes have been investigated. Amavadin was isolated from Amanita muscaria and oxidized with [NH(4)](2)[Ce(NO(3))(6)]. H(2)[Delta-V(S,S-HIDPA)(2)].3H(2)O, H(2)[Delta,Lambda-V(S,S-HIDPA)(2)].3H(2)O and their equivalent oxidized species have been synthesized and characterized spectroscopically. A combination of COSY, NOE, (1)H, (13)C-NMR and CD spectroscopy have been used to prove that the isolated natural product Amavadin consists of an almost equal mixture of the Delta- and Lambda-isomers of [V(S,S-HIDPA)(2)](2-).

Purpose: Activation of muscarinic receptors has been implicated in an increased risk of cataract after anticholinesterase treatment for glaucoma. The purpose of the present study was to determine the acetylcholine muscarinic receptor subtype(s) present in native human lens epithelial cells (NHLECs) and a human lens cell line, HLE-B3, and to compare the distribution in other ocular cells., Methods: Human lens cells were perfused with artificial aqueous humor (35 degrees C) after fura-2 incorporation, and calcium levels were measured using a fluorometric single-cell digital imaging system. Acetylcholine was the primary muscarinic agonist, and the receptor subtypes were elucidated by determining the relative effectiveness of pirenzepine and AF-DX 384 in blocking the agonist-induced response. The levels of expression of mRNA for the receptor subtypes M1 through M5 were determined by quantitative reverse transcription-polymerase chain reaction (QRT-PCR) using a sequence detection system (ABI Prism 7700; Perkin-Elmer, Foster City, CA). This was performed using total RNA extracted from native lens, retina, iris, and sclera and also cultured lens cells., Results: Acetylcholine induced a similar concentration-dependent increase in peak-amplitude cytosolic calcium in the range 100 nM to 100 microM in both native and HLE-B3 cells. However, the kinetics of the response waveforms to 30-second pulses of acetylcholine were different in the two cell types. At higher concentrations (> 1 microM), a second phase appeared in the HLE-B3 cells that was absent in the NHLEC response. The 50% inhibitory concentration (IC50) values for blockade of a 1 microM acetylcholine response by pirenzepine and AF-DX 384 were 30 nM and 230 nM, respectively, for NHLECs, and 300 nM and 92 nM, respectively, for HLE-B3 cells. The QRT-PCR data showed that more than 90% of the total muscarinic receptor mRNA from NHLEC was of M1 origin. In the HLE-B3 cells, however, more than 95% of the mRNA was of M3 origin. mRNA for M3 was also in greatest abundance in other eye tissues, although there was a significant contribution from M1 in iris and sclera., Conclusions: Both NHLECs and HLE-B3 cells express muscarinic receptors that produce significant changes in cytosolic calcium in response to acetylcholine. Both pharmacologic and QRT-PCR evidence shows that whereas the M1 subtype predominates in NHLECs, M3 is the major contributor in HLE-B3 cells. In all other eye tissues, M3 appears to be the major contributor. These data should be taken into account when choosing particular models to investigate cataract mechanisms and also when designing muscarinic agonists to treat glaucoma.

The development of the understanding of the co-ordination chemistry and the properties of Amavadin, the chemical form in which vanadium is accumulated by the Amanita genus of mushrooms, is reviewed.

Objectives: To describe current primary care sun protection advice for children and assess the effect on clinicians of an intervention to enhance their sun protection advocacy., Setting: Primary care practices caring for children in New Hampshire with special attention to clinicians serving 10 towns that were involved in a randomized controlled trial of the multicomponent SunSafe intervention involving schools, recreation areas, and primary care practices., Design/intervention: A statewide survey of all primary care clinicians serving children addressed their self-reported sun protection advocacy practices. Clinicians in 10 systematically selected rural towns were involved in the subsequent intervention study. The primary care intervention provided assistance to practices in establishing an office system that promoted sun protection advice to children and their parents during office visits., Main Outcome Measures: Sun protection promotion activities of primary care clinicians as determined by their self report, research assistant observation, and parent interviews., Results: Of 261 eligible clinicians responding to the statewide survey, about half provide sun protection counseling "most of the time" or "almost always" during summer well care visits. Pediatricians do so more often than family physicians. Clinicians involved in the intervention increased their use of handouts, waiting room educational materials, and sunscreen samples. Compared with control town parents, parents in intervention towns reported an increase in clinician sun protection advice., Conclusions: The SunSafe primary care intervention increased sun protection counseling activities of participating clinicians. A single-focus preventive service office system is feasible to include in community interventions to promote sun protection.

Actinyl ions (AnO(2)(+) and AnO(2)(2+)) are strongly complexed by CO(3)(2)(-) in aqueous solution, and tricarbonato complexes of UO(2)(2+) and NpO(2)(+), in particular, have been widely studied. Cyclic voltammetry shows that, whereas the Np(V)/Np(VI) couple is reversible in aqueous CO(3)(2)(-) solution, the analogous U couple is irreversible, suggesting significant conformational change on one-electron reduction of [UO(2)(CO(3))(3)](4)(-). EXAFS spectroscopy has been used to define the coordination geometry of the reduced species and shows that both axial and equatorial U-O distances increase on reduction, from 1.80 to 1.90 Å and from 2.43 to 2.50 Å, respectively. The basic geometry of the U complex is apparently unchanged by reduction so any conformational change which does occur must be subtle.

Fly agaric accumulate vanadium in the form of amavadin, whose structure has been elucidated (see picture for the crystal structure). Amavadin contains one V IV center coordinated to two (S,S)-hidpa 3- ligands (H 3 hidpa=2,2'-(hydroxyimino)dipropionic acid) through one η 2 -NO - group and two unidentate carboxylato groups from each ligand. The arrangement of the two η 2 -NO - groups leads to a chiral vanadium center, which has been characterized in both the Λ and Δ forms. The carboxylato groups of the amavadin anions bind cations, for example, Ca 2+ ions, and/or become involved in hydrogen bonding., (© 1999 WILEY-VCH Verlag GmbH, Weinheim, Fed. Rep. of Germany.)

A tetranuclear iron cluster is the principal component of the purple coatings produced by treating a mild steel surface with a salicylaldoxime corrosion inhibitor. This was shown by comparison of the spectroscopic data with those of the cluster [{Fe(salH)(HsalH)}4 ], which was obtained from FeCl3 and salicylaldoxime (H2 salH) and has a distorted tetrahedral arrangement of Fe(III) atoms coordinated by terminal (1-) and bridging (2-) salicylaldoximate ligands (the central core of the cluster is depicted)., (Copyright © 1999 WILEY-VCH Verlag GmbH, Weinheim, Fed. Rep. of Germany.)

Objective: We evaluated the impact of an intervention promoting sun protection behavior among children 2 to 11 years of age through schools and day care centers, primary care practices, and recreation areas., Methods: Ten towns in New Hampshire were paired, then assigned randomly to intervention or control status. The multicomponent SunSafe intervention was provided to children and caregivers through primary care practices, day care centers, schools, and beach recreation areas. Training support and materials were provided by the SunSafe project, but project staff had no direct contact with children or parents in providing the intervention. All intervention components promoted the same message: avoid the sun between 11 AM and 3 PM, cover up using hats and protective clothing, use sun block with a sun protection factor >/=15, and encourage sun protection among family and friends. The impact of the intervention was determined by observing children's sun protection behavior at the beach during baseline compared with 1 year later. The primary outcomes of interest were changes in the proportion of children per town using at least some sun protection and changes in the proportion of children fully protected. Children were clustered by town, with the town thus being the unit of analysis. The primary care practice component included one practice meeting for clinicians and staff at which project staff presented background on skin cancer and how to promote its prevention; a sun protection office system manual based on our previous work, which provided specific direction on how to share responsibility among office staff and clinicians in carrying out routines that promote sun protection; and educational posters, pamphlets, and self-adhesive reminder notes designed to enhance sun protection counseling. SunSafe removable tattoos and stickers were offered to children at well-child and illness visits during the summer months. Schools each received three project staff visits: a brief visit with the principal to describe the intervention and to answer questions; an in-service program to educate teachers about skin cancer and to introduce curricular materials; and help with one parent outreach program. Larger day care centers each received one project staff visit. An additional six smaller day care centers received curricular materials through the mail but no visits. Two similar sets of curricular materials were used, one for grade schools and the other for preschools and day care centers. Both emphasized the importance of sun protection rather than the danger of skin cancer. Materials emphasized dynamic activities modeled after the "Slip, Slop, Slap" and "SunSmart" programs and included new material developed to suit regional needs. Both manuals offered structured plans but also provided a variety of activities from which teachers could choose. Teachers agreed to devote a minimum of two class periods to these materials. For recreation areas, lifeguards in each of the intervention communities attended an in-service meeting, during which background about skin cancer prevention was presented by project staff. The project also provided displays about the ultraviolet (UV) light index and about sun protection to be posted at each beach. Subsequently, project staff called beach staff in each community each morning with the predicted UV index for the day to post on the display. Educational pamphlets about the UV index and free sun-block samples were available to beachgoers through the lifeguards. One brief follow-up visit by project staff was made to each beach area to provide reinforcement., Results: We observed 1930 children. Use of some sunscreen on at least one body area increased in all 5 intervention towns compared with paired control towns. In intervention towns, this mean proportion increased from 0.56 of those observed at baseline to 0.76 of those observed postintervention, with a minimal increase among control town children. (ABSTRACT TRUNCATED

Background: Perichondrial cutaneous grafts (PCCGs) are composite grafts comprised of skin and subjacent perichondrium. Animal models and preliminary clinical reports have shown that PCCGs are thicker, survive better and contract less than full-thickness skin grafts, and are simpler to perform that alternative reconstructive methods such as two-stage flaps., Objective: The applicability of PCCGs to surgical defects following Mohs surgery was investigated., Methods: PCCGs were used to reconstruct patients with surgical defects following Mohs excision of skin cancers from facial sites near free anatomic margins and in defects with exposed cartilage., Results: A series of cases is described, all with good to excellent cosmesis and function., Conclusion: PCCGs are useful in reconstruction of defects following Mohs excision of skin cancers.

Removal of the nail matrix is indicated in a variety of recalcitrant conditions in which the growth of the nail plate causes mechanical problems such as recurrent onychocryptosis or pain due to pinching or pressure. Before performing a matricectomy, the physician must consider the use of nonsurgical methods and the possible contraindications to surgery. Once the decision for matricectomy is made, the physician should then utilize the modality with the least morbidity and greatest convenience for the patient. A wide repertoire of methods of matricectomy is available for use; all methods, when performed properly, have similar high rates of cure.

A 39-year-old white female with multiple arteriovenous malformations of the left foot had had surgical ligation on her left dorsalis pedis artery at age 12 with no clinical improvement. She was placed into a leotard at age 16 and was able to function but would tire very easily and her left foot was twice the size of her right one. In May 1988, she had four large feeder vessels off the posterior tibial artery and two off the anterior tibial artery embolized by use of 1,000 microns PVA particles, occlusive spring coils, and detachable balloons. Her foot shrunk and is now smaller than her normal right foot; she does not tire, and she now wears a knee length stocking to help prevent other channels from opening.

The kinetics of inhibition of dopamine (3,4-dihydroxyphenethylamine) beta-mono-oxygenase by cyanide (CN-) and azide (N3-) ions have been investigated by using steady-state methods. Both anions show complex non-competitive-inhibition patterns with respect to ascorbate, suggestive of anion binding at two different sites on the oxidized enzyme. To further investigate this finding, e.p.r. titrations of CN- and N3- binding to the 63Cu-reconstituted enzyme were carried out. Addition of approx. 2 equiv. of CN- to copper elicits a new signal with g = 2.217, g = 2.025, A = 17.0 mT characteristic of a copper (II)-cyano complex. Simulations show that this signal accounts for half the copper (II) in the enzyme. The remainder of the enzyme-bound copper is expressed by a signal close to, but not identical with, that of native enzyme. Further addition of CN- induces a simultaneous decrease in intensity of both of these signals so that their 1:1 ratio is maintained. Binding of N3-, on the other hand, changes the e.p.r. spectrum to a form different from either that of the native or CN- -treated enzyme, and integrates to 100% of the copper in the enzyme (g = 2.252, g = 2.050, A = 16.5 mT). Resolved superhyperfine structure is apparent in the g region. N3- binding is also accompanied by the appearance of a broad charge-transfer band centred at 387 nm. Neither 9 nor 35 GHz e.p.r. spectra show evidence for more than one (non-interacting) species of Cu(II) in native enzyme and N3- derivatives. The binding and reactivity of CN-, on the other hand, argues against independent copper sites in the enzyme.

Sebaceous wax ester secretion rates were measured in six subjects on six occasions each, using absorption into bentonite clay and analysis of the collected lipid by quantitative thin-layer chromatography. On each occasion eight samples were collected, four from the left and four from the right side of the forehead in four successive intervals. The first two intervals, which totaled 14 h, were intended to deplete the follicular reservoir of sebum so that a constant rate of absorption could be obtained during the third and fourth intervals, which were 3 h each. Thin-layer analysis of each sample was done in triplicate. The data were examined using analysis of variance techniques to determine the reproducibility of the measurement method and to identify possible sources of variability. The intraclass correlation coefficient (rI) for all 432 post-depletion determinations was 0.80. The reproducibility was considerably better for three of the subjects (rI = 0.93) than for the other three (rI = 0.75). Variability within the latter three subjects did not seem to be attributable to lack of reproducibility in the thin-layer analysis. Real biological variability also seems unlikely considering the holocrine mechanism of sebum secretion. Therefore, the variability probably arises from non-representative collection of sebum into the bentonite absorbent.

A 64-year-old white male had extensive ulcerative cutaneous sarcoidosis of the lower extremities. After stabilization with methotrexate and prednisone therapy, the ulcers were debrided and covered with split-thickness skin grafts. An 80% graft acceptance was maintained 2 months later. Successful grafting of ulcerative sarcoidosis has not been reported previously.

Histiocytosis X (HX) is a rare disorder of histiocytic proliferation characterized by a broad spectrum of clinicopathologic disease. An unusual case of Letterer-Siwe disease (LSD) or subacute disseminated HX in a 71-year-old woman is presented. The patient had a 3-year history of splenomegaly before skin lesions developed. She presented to our clinic at 1.5 years later and the diagnosis of HX was made by skin biopsy. Topical nitrogen mustard (NM) therapy resulted in complete clearing of cutaneous lesions. Her condition was stable over the next 10 months. However, she subsequently suffered a rapid and fatal dissemination of her disease. Systemic treatment with prednisone, vinblastine sulphate, and suppressin A (SA) (a calf thymus derived hormone preparation that specifically induces suppressor T-cells) was ineffective. Characteristic histopathologic, immunohistochemical, and electron microscopic findings of HX are illustrated. A review of the adult cases of LSD and treatment options for HX are presented and discussed.

Amine oxidase from pig plasma (PPAO) has two bound Cu2+ ions and at least one pyrroloquinoline quinone (PQQ) moiety as cofactors. It is shown that recovery of activity by copper-depleted PPAO is linear with respect to added Cu2+ ions. Recovery of e.s.r. and optical spectral characteristics of active-site copper parallel the recovery of catalytic activity. These results are consistent with both Cu2+ ions contributing to catalysis. Further e.s.r. studies indicate that the two copper sites in PPAO, unlike those in amine oxidases from other sources, are chemically distinct. These comparative studies establish that non-identity of the Cu2+ ions in PPAO is not a requirement for amine oxidase activity. It is shown through the use of a new assay procedure that there are two molecules of PQQ bound per molecule of protein in PPAO; only the more reactive of these PQQ moieties is required for activity.

Certain patients with bronchial asthma can benefit, often greatly, from hypnotherapy. This report is based on a retrospective analysis of 121 asthmatic patients who were treated by hypnotherapy. Hypnotic techniques and treatment procedure are described. Of the total number, 21% had an excellent response to treatment, becoming completely free from asthma and requiring no drug therapy. A further 33% had a good response, with worthwhile decrease in frequency and severity of the attacks of asthma, or a decrease in drug requirements. About half of the 46% who had a poor response had a marked subjective improvement in general well-being. Statistical evaluation of the six variables (age, sex, result, trance depth, psychological factors and severity of the asthma) confirmed the clinical impression that the ability to go into a deep trance (closely associated with the youthfulness of the subject) gives the best possibility of improvement, especially if there are significant aetiological psychological factors present and the asthma is not severe. Subjective improvement in well-being and outlood is a potential outcome at all age levels, independent of severity of the illness or entranceability of the patient.

Potentiometric titrations, continuous wave EPR, and microwave power saturation measurements have been used to examine 8-Cu and 4-Cu forms of native dopamine beta-hydroxylase and its azide derivative. The formation curve for the binding of Cu2+ to the apoenzyme is best fit by assuming two independent binding sites per subunit, with pK' values of 8.90 and 7.35 at pH 5.0. On the other hand, only minor differences are observed in either continuous wave EPR spectra or power saturation behavior of the 8- and 4-Cu forms of the native enzyme or of its azide derivative. The intensity of the EPR spectra of all derivatives integrates to greater than 95% of the total copper, and the temperature dependence of P1/2 shows no evidence for any S = 1 state of the copper ions in the enzyme. These results suggest a lower limit of ca. 7 A for the separation between the two copper ions per subunit and thus rule out a type 3 site in the oxidized enzyme. The data are most consistent with Cu(II) sites consisting of two or three N (imidazole) and one or two O donor ligands in the coordination sphere. The similarity in EPR spectra and power saturation of 8- and 4-Cu derivatives suggests that the difference in Cu-binding constants may be due either to differences in the identity of an axial ligand or to solvation effects in the active site.