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Background The disposition of the long-acting opioid methadone, used to prevent opiate withdrawal and treat short- and long-lasting pain, is highly variable. Methadone undergoes N-demethylation to the primary metabolite 2-ethyl-1,5-dimethyl-3,3-diphenylpyrrolinium (EDDP), catalyzed in vitro by intestinal, hepatic, and expressed cytochrome P450 (CYP) 3A4. However, the role of CYP3A4 in human methadone disposition in vivo is unclear. This investigation tested the hypothesis that CYP3A induction (or inhibition) would increase (or decrease) methadone metabolism and clearance in humans. Methods Healthy volunteers were studied in a randomized, balanced, 4-way crossover study. They received intravenous (IV) midazolam (to assess CYP3A4 activity) and then simultaneous oral deuterium-labeled and IV unlabeled methadone after pretreatment with rifampin (INN, rifampicin) (hepatic/intestinal CYP3A induction), troleandomycin (hepatic/intestinal CYP3A inhibition), grapefruit juice (selective intestinal CYP3A inhibition), or nothing. Methadone effects were measured by dark-adapted pupil diameter. CYP isoforms catalyzing methadone metabolism by human liver microsomes and expressed CYPs in vitro were also evaluated. Results Methadone had high oral bioavailability (70%) and low intestinal (22%) and hepatic (9%) extraction, and there was a significant correlation (r = 0.94, P < .001) between oral bioavailability and intestinal (but not hepatic) availability. Rifampin decreased bioavailability and oral and IV methadone plasma concentrations and increased IV clearance (4.42 ± 1.00 mL · kg−1 · min−1 versus 1.61 ± 0.67 mL · kg−1 · min−1, P < .05) and oral clearance (8.50 ± 3.68 mL · kg−1 · min−1 versus 2.05 ± 0.92 mL · kg−1 · min−1, P < .05), EDDP/methadone area under the curve (AUC) ratios, EDDP formation clearances, and hepatic extraction (0.27 ± 0.06 versus 0.09 ± 0.04, P < .05). Troleandomycin and grapefruit juice decreased the EDDP/methadone AUC ratio after oral methadone (0.17 ± 0.10 and 0.14 ± 0.06 versus 0.27 ± 0.20, P < .05) but not IV methadone and had no effect on methadone plasma concentrations, IV clearance (1.29 ± 0.41 mL · kg−1 · min−1 and 1.48 ± 0.55 mL · kg−1 · min−1) or oral clearance (2.05 ± 1.52 mL · kg−1 · min−1 and 1.89 ± 1.07 mL · kg−1 · min−1), or other kinetic parameters. There was no correlation between methadone clearance and hepatic CYP3A4 activity. Pupil diameter changes reflected plasma methadone concentrations. In vitro experiments showed a predominant role for both CYP3A4 and CYP2B6 in liver microsomal methadone N-demethylation. Conclusion First-pass intestinal metabolism is a determinant of methadone bioavailability. Intestinal and hepatic CYP3A activity only slightly affects human methadone N-demethylation but has no significant effect on methadone concentrations, clearance, or clinical effects. Greater rifampin effects, compared with troleandomycin and grapefruit juice, on methadone disposition suggest a major role for intestinal transporters and for other CYPs, such as CYP2B6. Interindividual variability and drug interactions affecting intestinal transporter and hepatic CYP3A and CYP2B6 activity may alter methadone disposition. Clinical Pharmacology & Therapeutics (2004) 76, 250–269; doi: 10.1016/j.clpt.2004.05.003

Ghrelin, identified in the gastric mucosa has been involved in control of food intake and growth hormone (GH) release but little is known about its influence on gastric secretion and mucosal integrity. The effects of ghrelin on gastric secretion, plasma gastrin and gastric lesions induced in rats by 75% ethanol or 3.5 h of water immersion and restraint stress (WRS) were determined. Exogenous ghrelin (5, 10, 20, 40 and 80 μg/kg i.p.) increased gastric acid secretion and attenuated gastric lesions induced by ethanol and WRS and this was accompanied by the significant rise in plasma ghrelin level, gastric mucosal blood flow (GBF) and luminal NO concentrations. Ghrelin-induced protection was abolished by vagotomy and attenuated by suppression of COX, deactivation of afferent nerves with neurotoxic dose of capsaicin or CGRP8–37 and by inhibition of NOS with l -NNA but not influenced by medullectomy and administration of 6-hydroxydopamine. We conclude that ghrelin exerts a potent protective action on the stomach of rats exposed to ethanol and WRS, and these effects depend upon vagal activity, sensory nerves and hyperemia mediated by NOS-NO and COX-PG systems.

Previous investigations have demonstrated that capsaicin-sensitive sensory nerves are involved in the development of hypertension in some rat models of hypertension. To determine the role played by calcitonin gene-related peptide (CGRP; the predominant neurotransmitter in capsaicin-sensitive sensory nerves) in a rat model of neurogenic hypertension, in which hypertension was induced by injecting 50 A lo f 10% phenol in the lower pole of the left kidney, systolic blood pressure (SBP) was monitored by the tail-cuff method throughout the experiment. Fifteen days after injection of phenol, mean arterial pressure (MAP), concentrations of CGRP in the plasma, the expression of CGRP mRNA in dorsal root ganglia (DRG) and CGRP content in laminae I and II of the spinal cord were measured. SBP was significantly increased 5 days after the intrarenal injection of phenol (164F7 mm Hg, p

Purpose : To study the efficacy of several subjective and objective methods of accommodation measurement in normal prepresbyopic and presbyopic populations to identify appropriate methods for measuring the outcome of accommodative restorative procedures. Setting : University of Houston, College of Optometry, Houston, Texas, USA. Methods : Thirty-one normal subjects with a mean age of 43.7 years (range 31 to 53 years) participated. Accommodation was measured monocularly using 3 subjective approaches—the push-up test, minus lenses to blur, and a focometer—and 2 approaches measured with a Hartinger coincidence refractometer, in which accommodation was stimulated with minus lenses to blur and topical pilocarpine 6%. Results : The push-up method overestimated accommodative amplitude relative to objective measures in 28 subjects. Two subjective methods, minus lenses to blur and the focometer, produced comparable results, but with lower amplitudes in younger subjects and higher amplitudes in older subjects compared with objective methods. Comparable results were obtained when accommodation was stimulated in 1 of 2 ways and measured with the Hartinger. Pilocarpine elicited stronger accommodative responses than distance blur for subjects with low accommodative amplitudes. Pilocarpine 6% produced stronger responses in subjects with light irides than in those with dark irides. Conclusions : Hartinger-measured accommodation provides more realistic measurement of accommodative amplitude than the subjective methods tested, especially in the presbyopic population. In presbyopic subjects, the subjective tests resulted in accommodative amplitudes up to 4.0 diopters greater than those measured with objective tests. Measurements of accommodative amplitude are best achieved with objective methods to stimulate and measure accommodation.

This is an investigation of the effect of changing the pupil diameter on the P1 amplitude and latency of the multifocal electroretinogram (mfERG). MfERGs were recorded using a custom built wide field electrophysiological system. An array of 61 empirically scaled hexagons was used to stimulate the visual field. The duration of overall recording period was 8 min, segmented into 16 intervals each lasting 30 s. A combination of mydriatics and miotics were used to pharmacologically alter the pupil size and diameters between 1 and 10 mm were measured. There was a reduction in mfERG P1 amplitude in some cases greater than 50% (mfERG P1 amplitude 53 nV at 8 mm to 25 nV at 1 mm), with a change in pupil diameter of 7 mm. The mfERG P1 latency increased in some cases by as much as 8 ms in the central 40 degrees (mfERG P1 latency 39 ms at 8 mm to 47 ms at 1 mm). These results suggest that pupil size has significant effects on mfERG P1 amplitude and latency.

Purpose: To determine the effect of intracameral injection of preservative-free lidocaine 1% and carbachol 0.01% on corneal endothelial cells of rabbits. Setting: Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan. Methods: Forty eyes of 20 New Zealand White rabbits were divided into 2 equal groups. In the first group, 1 eye was injected with 0.02 mL of preservative-free lidocaine 1% and the fellow eye was injected with 0.02 mL of normal saline as a control. In the second group, 1 eye was injected with 0.02 mL of carbachol 0.01% and the fellow eye was injected with 0.02 mL of normal saline. Specular microscopy was used to evaluate corneal endothelial cell loss and corneal thickness 1 week and 1 month postinjection. For morphologic studies, corneal buttons were excised and stained with alizarin red with trypan blue. Scanning electron microscopy (SEM) examination was performed. Results: Specular microscopy revealed no significant endothelial cell loss and normal endothelial thickness with the intracameral injection of preservative-free lidocaine 1% and carbachol 0.01% compared with the control eye. Alizarin red with trypan blue stain and SEM examinations revealed smooth, distinct, and intact intercellular borders and normal viability of corneal endothelial cells in both groups. Conclusions: Intracameral injections of preservative-free lidocaine 1% and carbachol 0.01% do not produce morphologic and functional changes in the corneal endothelial cells of rabbits.

Although a posterior chamber phakic intraocular lens provides effective refractive correction of high myopia and hyperopia, mechanical contact between the implantable contact lens (ICL) and the crystalline lens and inadequate aqueous circulation in the prelenticular space could cause subcapsular opacification. To assess whether and to what extent such mechanical contact occurs, changes in the distance between the STAAR Collamer ICL and the crystalline lens under various conditions were investigated.Open pilot study.Thirteen eyes of 11 myopic and 2 hyperopic patients with a mean age of 38 years (range, 19-53 years) were examined at least 6 months after ICL implantation.A noninvasive, high-resolution biometry technique, partial coherence interferometry, was used to measure distance changes between the ICL and the crystalline lens during subjective accommodation, after instillation of pilocarpine, and under changing light conditions.Mean distance changes from the posterior corneal surface to the ICL, from the posterior corneal surface to the anterior surface of the crystalline lens, and the distance between the ICL and the crystalline lens.In the nonaccommodated state, the mean distance between the ICL and the crystalline lens was 457 microm (range, 123-924 microm). During subjective accommodation, a significant (P0.01) decrease and, after topical application of pilocarpine, a nonsignificant (P=0.35) decrease of anterior chamber depth was accompanied by a nonsignificant (P = 0.71) reduction of the ICL-crystalline lens distance. Under photopic conditions, a significant mean reduction (P0.01) of the ICL-crystalline lens distance of -28 microm (range, -16 to -188 microm) was observed.Partial coherence interferometry biometry enabled noninvasive high-precision investigation of ICL dynamics. No significant changes between the ICL and the crystalline lens were detected during subjective accommodation and after application of pilocarpine. However, under photopic conditions, with constriction of the pupil, the distance between the ICL and the crystalline lens was significantly reduced. This mechanism might cause inadequate aqueous circulation in the prelenticular space and might be one of the causes of subcapsular opacification in some of the eyes after ICL implantation.

OBJECTIVE: To evaluate the efficacy and side effects of oral pilocarpine for the treatment of ocular symptoms in patients with primary Sjogren's syndrome (SS). METHODS: A 12 week, single centre, randomised controlled study was performed. Twenty nine patients were randomly assigned to receive oral pilocarpine (5 mg twice a day), 28 only artificial tears, and 28 inferior puncta occlusion. Patients receiving oral pilocarpine and those with inferior puncta occlusion also received artificial tears. Patients were evaluated at baseline and throughout the study for their subjective global assessment of dry eyes and for their objective assessment of dry eyes (Schirmer's-I test, rose bengal test, and imprint test). RESULTS: Patients taking oral pilocarpine had significant improvement in subjective global assessment of dry eyes, as was evaluated by improvement of >55 mm on a visual analogue scale (VAS) for responses to the eye questionnaire, compared with patients treated with artificial tears (p55 mm on a VAS. Additionally, an improvement of rose bengal staining was noted, but an increase in tear production, as measured by the Schirmer-I test, was not substantiated. Ann Rheum Dis

To determine whether the efficacy of entry and action of antisense oligonucleotides (AS-ODN) on hematopoietic stem cells in vitro could be improved by the addition of polyethylene glycol (PEG), a molecule of PEG was bound to AS- or sense-acetylcholinesterase (AS-ACHE or S-ACHE). The introduction of 0.1-0.5 microM PEG-AS-ACHE or 0.5 microM AS-ACHE into methylcellulose bone marrow (BM) cultures produced a doubling in number of colony-forming unit-granulocyte-erythrocyte-macrophage-megakaryocyte (CFU-GEMM) and a 5-fold increase in cell number of the PEG-ODN. Further increase in concentration of the PEG-ODN reduced colony numbers. PEG-AS-ACHE induced higher colony numbers and greatly increased megakaryocyte (MK) formation when compared with PEG and AS-ACHE added separately to the culture. In addition, differentials of the CFU-GEMMs indicated there was a direct relationship between MK number and PEG-AS-ACHE concentration. Under these culture conditions, 5 microM PEG alone gave control values of CFU-GEMM. On addition of FITC-PEG-AS-ACHE to the cell cultures, using confocal microscopy, the nuclei of both early and mature MKs were labeled specifically, whereas all other cellular nuclei were negative to the stain. The use of PEG-AS-ODN, affording specific delivery of AS-ODN to target cells, increased cell proliferation, and enhanced ODN uptake, may be of potential importance in stem cell expansion for BM transplantation and gene therapy.

Solutes concentration in the aqueous humor of the eye is highest near the ciliary processes in the posterior chamber (p.c.) and lowest near the trabecular meshwork in the anterior chamber (a.c.). The high osmotic gradient across the semipermeable iris causes it to easily move forward at this location and occasionally occlude the iridocorneal angle. Vector analysis of the forces generated by miosis helps explain on osmotic grounds why it reduces the intraocular pressure, while at the same time the analysis elucidates the reason for the higher frequency of angle closure glaucoma when the anterior chamber is shallow.

Adrenomedullin (AM) is a potent vascular wall-derived vasorelaxing peptide which induces the release of nitric oxide (NO). To explore the role of endogenous AM in vascular function, we examined the effects of acetylcholine (ACh), AM, and AM receptor antagonists [AM (22-52), and calcitonin gene-related peptide (CGRP) (8-37)] on the isometric tension of aortic rings isolated from AM transgenic (TG) and knockout (KO) mice and wild type littermates (WT). ACh and AM caused a dose-dependent reduction of the isometric tension of aortic rings, but the degree of vasodilatation was smaller in TG than in KO or WT (% delta tension [10(-6) mol/l ACh]: KO -69 +/- 10%, WT -39 +/- 8%, TG -29 +/- 1%, p0.01). On the other hand, N(G)-nitro-L-arginine methyl ester, an NO synthase inhibitor, induced greater vasoconstriction in TG (% delta tension 10(-5)mol/l: KO +78 +/- 16%, WT +99 +/- 27%, TG +184 +/- 20%, p0.01), whereas E-4021, a cyclic guanosine monophosphate (cGMP)-specific phosphodiesterase inhibitor, caused greater vasodilation in TG mice. Both AM antagonists increased tension in TG to a greater extent than in KO or WT mice (% delta tension [10(-6) mol/l CGRP (8-37)]: KO +24 +/- 5%, WT +51 +/- 6%, TG +75 +/- 7%, p0.01). Endothelial denudation of the aorta diminished the vasoconstriction caused by the AM antagonists. In conclusion, the amounts of AM expressed in the aortic endothelium influenced baseline NO release. AM antagonists increased vascular tone in WT as well as in TG, suggesting that endogenous AM plays a physiological role in the regulation of aortic tone.

Objective: This study was performed to investigate the influence of calcitonin gene-related peptide (CGRP) and substance P (SP) on the intra-urethral pressure in female rats, as both these neuropeptides have been demonstrated to occur in nerve fibres throughout all layers of the intrinsic external urethral sphincter of the rat. Material and Methods: Both CGRP and SP were administered intra-arterially relatively close to the bladder in ketamine-anaesthetized female rats. The maximum urethral pressure (MUP) was recorded continuously using a 4 F microtip single-transducer catheter. Results: Pronounced effects on the intra-urethral pressure were found with both CGRP and SP. CGRP at the maximum dose given (10 mug) induced an immediate, pronounced, long-lasting decrease in pressure from 28 +/- 4 to 10 +/- 2 cmH 2 O, amounting to 65% of the MUP. SP (10 mug) instead induced a forceful, phasic, peak-like contractile response with a 170% increase in urethral pressure from 33 +/- 6 to 87 +/- 6 cmH 2 O of the initial control level of MUP. Antagonists to CGRP and SP did not induce any pressure changes per se. Conclusion: These results indicate that both CGRP and SP are of importance for the peripheral motor regulation of the external urethral sphincter, and hence possibly also of physiological importance for lower urinary tract function during essential parts of the micturition cycle. (Less)

Objetivo: Estudar os efeitos da redução do diâmetro pupilar nos resultados da perimetria de freqüência dupla em um grupo de voluntários normais. Métodos: Dezoito voluntários saudáveis participaram do estudo. Realizou-se perimetria de freqüência dupla no olho direito de cada participante (estratégia "Full Threshold" C-20). Para a segunda sessão de perimetria, instilou-se uma gota de pilocarpina 2% no olho direito dos voluntários e refez-se o exame após 60 minutos. Resultados: Sessenta minutos após adição de pilocarpina 2% houve redução significante do diâmetro pupilar de 4,22 ± 0,17 mm para 1,55 ± 0,51 mm (p0,05). Conclusão: A alteração do diâmetro pupilar está associada a redução significativa dos limiares de sensibilidade dentro dos 20º centrais do campo testado pela perimetria de freqüência dupla. Estes resultados sugerem que seja importante manter o diâmetro pupilar constante em exames seriados.Purpose: To study the effects of pupillary constriction on frequency doubling perimetry in a group of normal subjects. Methods: Eighteen healthy volunteers participated in the study. Only one eye per patient (right eye) underwent frequency doubling perimetry (Full Threshold C-20 strategy). For the second session, one drop of 2% pilocarpine was administered to the volunteers' right eye and the examination was repeated after 60 minutes. Results: Sixty minutes after administration of 2% pilocarpine, there was a significant reduction of the pupillary diameter from 4.22 ± 0.17 mm to 1.55 ± 0.51 mm (p

• Diplopia in the form of acute esotropia may manifest in up to 30% of individuals undergoing heroin withdrawal.

REVIEẂS AIM: When a phacoemulsification, a filtration surgery or a combined surgery are necessary, questions about the convenience of continuing certain antiglaucomatous drugs could appear. The aim of this review article is to unify criteria that will guide daily clinical practice and including the developing algorithms of action in the preoperative and postoperative periods of filtration surgery and/or cataract surgery.In the preoperative period of cataract surgery, the use of non-steroidal anti-inflammatory drugs is at the discretion of the surgeon, with the monodose presentation being recommended. The suspension of prostaglandines a fewdays before the surgery should be considered. Preservative-free drugs ensure a better recovery of the ocular surface (OS) after cataract surgery. Once all modifying factors of the intraocular pressure (IOP) have been removed, baseline IOP should be evaluated again, choosing preservative-free antiglaucomatous drugs when needed. The use of preservative-free ocular antihypertensive drugs and steroids in the preoperative period of glaucoma surgery reduces the risk of surgical failure. The interruption of prostaglandines is recommended. In the postoperative period of glaucoma surgery, steroids are the anti-inflammatory treatment of choice, the preservative-free ones being preferred. When reintroducing antiglaucomatous treatment, preservatives should be avoided to prevent scarring. The appropriate perioperative management of patients with glaucoma is essential to obtain a correct control of IOP, improve the situation of the OS, prevent complications and improve the result of the filtration surgery and cataract surgery.this protocol aims to unify the different lines of action in order to decrease the incidence of adverse events and maximize the surgical outcome.

11 pags.; 7 figs., Objective To quantify 3-dimensionally the anterior segment geometry, biometry, and lens position and alignment in patients before and after implantation of the Crystalens-AO (Bausch & Lomb, Rochester, NY) accommodating intraocular lens (A-IOL). Design Prospective, observational study. Participants Ten patients (20 eyes) with cataract before and after implantation of the Crystalens-AO A-IOL. Methods Custom full anterior segment 3-dimensional (3-D) spectral optical coherence tomography (OCT) provided with quantification tools was used to image the cornea, iris, and natural lens preoperatively and intraocular lens postoperatively. Measurements were obtained under phenylephrine preoperatively and under natural viewing conditions and phenylephrine (for accommodative efforts ranging from 0 to 2.5 diopters [D]) and pilocarpine postoperatively. Main Outcome Measures Three-dimensional quantitative anterior segment images, corneal geometry and power, anterior chamber depth (ACD), lens thickness, pupil diameter, A-IOL shift with accommodative effort or drug-induced accommodation, and A-IOL alignment. Results Crystalline lens and IOLs were visualized and quantified 3-dimensionally. The average ACD were 2.64±0.24 and 3.65±0.35 mm preoperatively and postoperatively (relaxed state), respectively, and they were statistically significantly correlated (although their difference was not statistically correlated with lens thickness). The A-IOL did not shift systematically with accommodative effort, with 9 lenses moving forward and 11 lenses moving backward (under natural conditions). The average A-IOL shift under stimulated accommodation with pilocarpine was -0.02±0.20 mm. The greatest forward shift occurred bilaterally in 1 patient (-0.49 mm in the right eye and -0.52 mm in the left eye, under pilocarpine). The high right/left symmetry in the horizontal tilt of the crystalline lens is disrupted on IOL implantation. Accommodative IOLs tend to be slightly more vertically tilted than the crystalline lens, with increasing tendency with accommodative effort. Two subjects showed postoperative IOL tilts >9 degrees. Changes in pupillary diameter correlated with pilocarpine-induced A-IOL axial shift. Intermediate accommodative demands (1.25 D) elicited the greater shifts in axial A-IOL location and tilt and pupil diameter. Conclusions Quantitative 3-D anterior segment OCT allows full evaluation of the geometry of eyes implanted with A-IOLs preoperatively and postoperatively. High-resolution OCT measurements of the Crystalens 3-D positioning revealed small (and in many patients backward) A-IOL axial shifts with both natural or drug-induced accommodation, as well as tilt changes with respect to natural lens and accommodative effort. © 2014 by the American Academy of Ophthalmology., Financial Disclosure(s): Spanish patent: P201130685 (S.O. and S.M.). The research leading to these results has received funding from the European Research Council under the European Union's Seventh Framework Programme(FP7/2007-2013) / ERC Grant Agreement n° 294099. Also, it has been suported by FIS2011-25637(Spanish Goverment Grant) and the Consejo Superior de Investigaciones Científicas “Junta para la Ampliación de Estudios (JAE Pre to J.B).

To determine whether RhoA isoprenylation (geranylgeranylation) is required for agonist-induced actin cytoskeleton reorganization (measured by an increase in the filamentous F- to monomeric G-actin ratio), human airway smooth muscle cells were treated for 72 h with inhibitors of geranylgeranyltransferase I. Geranylgeranyltransferase inhibitor (GGTI)-2147 or -286 pretreatment completely blocked the increase in the F- to G-actin fluorescence ratio when cells were stimulated with lysophosphatidic acid (LPA), endothelin, or carbachol. In contrast, LPA or endothelin induced actin cytoskeletal reorganization in cells treated with farnesyltransferase inhibitor (FTI)-277 to inactivate Ras. Forskolin-induced adenylyl cyclase activity was inhibited by carbachol in GGTI-2147-pretreated cells, demonstrating that the effect of geranylgeranyltransferase I inhibition on stress fiber formation was not due to uncoupling of signaling between the heterotrimeric Giprotein (the Gγ subunit is isoprenylated) and distal effectors. These results demonstrate that selective GGTIs can inhibit agonist-induced actin reorganization.

Thermoreversible gels formed in situ by aqueous solutions of an enzyme-degraded xyloglucan polysaccharide were evaluated as sustained release vehicles for the ocular delivery of pilocarpine hydrochloride. In vitro release of pilocarpine from gels formed by warming xyloglucan sols (1.0, 1.5 and 2.0% w/w) to 34 degrees C followed root-time kinetics over a period of 6 h. The miotic responses in rabbit following administration of xyloglucan sols were compared with those from in situ gelling Pluronic F127 sols and from an aqueous buffer solution containing the same drug concentration. Sustained release of pilocarpine was observed with all gels, the duration of miotic response increasing with increase of xyloglucan concentration. The degree of enhancement of miotic response following sustained release of pilocarpine from the 1.5% w/w xyloglucan gel was similar to that from a 25% w/w Pluronic F127 gel.

The calcitonin-gene receptor peptide (alphaCGRP) receptor is present in high levels in the caudal striatum of the rat. Previous behavioural experiments have highlighted a possible correlation between alphaCGRP-mediated effects and the dopaminergic system. In this study, we examined the effect of alphaCGRP on K(+)-evoked [(3)H]-dopamine release in a slice preparation of the rat caudal striatum. The unstimulated release of [(3)H]-dopamine was not affected by alphaCGRP. However, alphaCGRP increased the release of [(3)H]-dopamine evoked by K(+) (30 mM) in a concentration-dependent manner. The stimulatory effect of alphaCGRP was blocked by the CGRP1 antagonist hCGRP(8-37) (without effect on its own). The stimulatory effect of 1 microM alphaCGRP was blocked by dizocilpine (MK-801), suggesting that excitatory transmission is involved in mediating the facilitated release. This study suggests that the peptide alphaCGRP, modulates dopamine release in the rat caudal striatum probably indirectly via glutamatergic transmission.

The neuropeptide calcitonin gene-related peptide (CGRP) is a potent microvascular vasodilator in rat skin and effects are antagonised by CGRP(8-37). In this study, CGRP(8-37) significantly (P0.05) inhibited the time-dependent (3-5 h) increase in skin blood flow measured in the anaesthetised rat, after intradermal administration of the inflammatory cytokine interleukin-1beta (3 pmol/site), indicating the involvement of CGRP1 receptors. The CGRP-related peptide adrenomedullin (ADM) is also a potent vasodilator in rat skin, with effects antagonised by CGRP(8-37). We show that ADM mRNA expression is increased in rat skin after treatment with IL-1beta and that the IL-1beta-induced blood flow is blocked by a selective ADM antibody (P0.05). Thus ADM is expressed locally in the inflamed cutaneous microvasculature where it can, in addition to, or as an alternative to CGRP, contribute to IL-1beta-induced vasoactive effects.

To determine the ocular hypotensive mechanism underlying the additivity of latanoprost and pilocarpine.This randomized, double-masked study included 30 patients with ocular hypertension on no ocular medications for at least 3 weeks. On each of six visits to the clinic, measurements were taken of aqueous flow and outflow facility by fluorophotometry, intraocular pressure by tonometry, and episcleral venous pressure by venomanometry. Uveoscleral outflow was calculated. Clinic visits were scheduled on baseline day; on day 8 of four times daily pilocarpine (2%) to one eye and vehicle to the other; on day 8 of continued pilocarpine/vehicle treatment plus latanoprost (0.005%) once daily to both eyes; after a 3-week washout period; on day 8 of once-daily latanoprost to one eye and vehicle to the other; and on day 8 of continued latanoprost/vehicle treatment plus pilocarpine four times a day to both eyes. Drug-treated eyes were compared with contralateral vehicle-treated eyes and with baseline day by paired t tests. Combined pilocarpine and latanoprost-treated eyes were compared with individual drug-treated eyes and with baseline day using the Bonferroni test.Compared with baseline, pilocarpine reduced intraocular pressure from 18.9 to 16.2 mm Hg (P =.001) and increased outflow facility from 0.18 to 0.23 microl per minute per mm Hg (P =.03). No other parameters were affected. Adding latanoprost further reduced intraocular pressure to 13.7 mm Hg (P.001) and increased uveoscleral outflow from 0.82 to 1.36 microl per minute (P =.02). Latanoprost alone reduced intraocular pressure from 17.6 to 14.3 mm Hg (P.0001) and increased uveoscleral outflow from 0.89 to 1.25 microl per minute (P =.05). Adding pilocarpine to the latanoprost treatment further reduced intraocular pressure to 12.7 mm Hg (P.001) and increased outflow facility from 0.21 to 0.30 microl per minute per mm Hg (P =.03).Latanoprost and pilocarpine predominantly increase uveoscleral outflow and outflow facility, respectively, when given alone. These drugs are additive because pilocarpine does not inhibit the uveoscleral outflow increase induced by latanoprost.

Objective: Xerostomia is a frequent and potentially debilitating toxicity of radiotherapy (XRT) for cancers of the head and neck. This report describes the use of acupuncture as palliation for such patients. Methods and Materials: Eighteen patients with xerostomia refractory to pilocarpine therapy after XRT for head and neck malignancy were offered acupuncture as palliation. All patients are without evidence of cancer recurrence at the primary site. Acupuncture was provided to three auricular points and one digital point bilaterally, with electrostimulation used variably. The Xerostomia Inventory (XI) was administered retrospectively to provide an objective measure of efficacy. Results: Acupuncture contributed to relief from xerostomia to varying degrees. Palliative effect as measured by the XI varied from nil to robust (pre- minus post- therapy values of over 20 points). Nine patients had benefit of over 10 points on the XI. Conclusions: Acupuncture reduces xerostomia in some patients who are otherwise refractory to best current management. © 2001 Elsevier Science Inc. Acupuncture, Head and neck neoplasms, Radiation therapy, Xerostomia.

We applied calcitonin gene-related peptide (CGRP) by continuous perfusion of the extrajunctional surface of the adult rat soleus muscle in vivo. We obtained this through a fine polyethylene catheter connected to an Alzet pump implanted in the animal. The perfusion induced a local acetylcholine receptor accumulation in the membrane of the muscle fibres starting with a delay of one to two days, provided a chronic conduction block of soleus innervation was concomitantly present. The effect was prominent, being higher than that following denervation. The lack of acetylcholine receptor accumulation observed in sham perfused animals and the co-administration of CGRP and its competitive antagonist peptide, hCGRP(8-37), eliminates the possibility that the response to CGRP application represents an inflammatory reaction to foreign bodies instead of a specific effect of the peptide. We suggest that CGRP may act on the extrajunctional membrane of muscle fibres to help induce acetylcholine receptor accumulation after appropriate receptors for the peptide are re-expressed due to muscle paralysis. Whilst this is compatible with a role of CGRP in synaptogenesis, a recent study showed that alpha-CGRP(-/-) mutant mice have normal neuromuscular junction development. However, given the redundancy of factors involved in acetylcholine receptor accumulation, further experiments on multiple knock-outs need to be performed before a final conclusion is reached about the physiological significance of CGRP.

The aim of the present contribution was to develop a functional three-dimensional tissue construct to study ocular permeation of pilocarpine hydrochloride from different formulations. The in vitro model was compared to excised bovine cornea. Modified Franz cells were used to study the transcorneal permeability. Analysis was performed by reversed-phase high-performance liquid chromatography. Comparisons of the permeation rates through excised bovine cornea and the in vitro model show the same rank order for the different formulations. The permeation coefficient, K P , obtained with the cornea construct, is about 2–4-fold higher than that from excised bovine cornea. It is possible to reconstruct bovine cornea as an organotypic culture and also to use this construct as a substitute for excised bovine cornea in drug permeation studies in vitro.

During recent decades consumption of antiglaucoma drugs has shown a steady increase in the Nordic countries. The cost increase can partly be explained by the use of newer, more expensive drugs. The increase in daily doses, however, is relatively much larger than the corresponding ageing of the population. There are large national and regional differences. Measured in daily defined doses (DDD) the consumption of glaucoma medication is almost three times as high in Sweden as in Denmark. Within all the Nordic countries there exists up to more than two-fold regional differences between high- and low-consuming counties. The new classes of IOP-lowering drugs have been very quickly adopted. Statistical data indicate that adding new drugs is more common than switching to a new therapy.

The common (A. antarcticus), northern (A. praelongus) and desert (A. pyrrhus) death adders are species belonging to the Acanthophis genus. The present study compared some pharmacological aspects of the venoms of these species and examined the in vitro efficacy of death adder antivenom. Neurotoxicity was determined by the time to produce 90% inhibition (t(90)) of indirect (0.1 Hz, 0.2 ms, supramaximal voltage) twitches in the chick biventer cervicis nerve-muscle (3-10 microg/ml) and mouse phrenic nerve-diaphragm (10 microg/ml) preparations. A. praelongus venom was significantly less neurotoxic than A. antarcticus venom but was not significantly different from A. pyrrhus venom. In the biventer muscle, all three venoms (3-10 microg/ml) abolished responses to exogenous ACh (1 mM) and carbachol (20 microM), but not KCl (40 mM), indicating activity at post-synaptic nicotinic receptors. All venoms (30 microg/ml) failed to produce significant inhibition of direct twitches (0.1 Hz, 2.0 ms, supramaximal voltage) in the chick biventer cervicis nerve-muscle preparation. However, A. praelongus (30 microg/ml) venom initiated a significant direct contracture of muscle, indicative of some myotoxic activity. The prior (10 min) administration of death adder antivenom (1 unit/ml), which is raised against A. antarcticus venom, markedly attenuated the twitch blockade produced by all venoms (10 microg/ml). Administration of antivenom (1.5 units/ml) at t(90) markedly reversed, over a period of 4 h, the inhibition of twitches produced by A. praelongus (3 microg/ml, 72+/-6% recovery) and A. pyrrhus (3 microg/ml, 51+/-9% recovery) but was less effective against A. antarcticus venom (3 microg/ml, 22+/-7% recovery). These results suggest that all three venoms contain postsynaptic neurotoxins. Death adder antivenom displayed differing efficacy against the in vitro neurotoxicity of the three venoms.

A 2.5-year-old domestic shorthair cat was evaluated because of dysphagia and weight loss of 4 weeks' duration. MIld blepharospasm and conjunctival hyperemia were evident in both eyes, oral mucous membranes were tacky on palpation, and salivary glands were enlarged. Results of a Schirmer tear test were 0 mm/min for both eyes. Administration of atropine did not cause salivation or caused secretion fo thick rope-like saliva. Examination of biopsy specimens of salivary glands revealed a plasmacytic infiltrate. Sjögren's syndrome (SS) was diagnosed. Oral administration of prednisone was instituted but was discontinued after a minimal positive response was evident 6 weeks after initiation of treatment. Palliative treatment with a 6% solution of pilocarpine 4 to 5 times/d, cyclosporine, hylan A, and neomycin-polymyxin-bacitracin ophthalmic ointment resulted in clinical improvement in the cat. Although reported rarely in animals, SS may be more common than currently is recognized. Most treatment regimens for SS are aimed at alleviating clinical signs.

The purposes of the present study were i) to determine whether neuropeptides induce the nasal obstruction in guinea pigs, and ii) to examine the possible involvement of neuropeptides in allergic nasal obstruction. The decrease in nasal cavity volume was determined by acoustic rhinometry as an index of nasal obstruction. In non-sensitized guinea pigs, substance P (SP), neurokinin A (NKA) and calcitonin gene-related peptide (CGRP) caused the nasal obstruction 10 to 30 min after their intranasal application. LY303870 (1 mg/kg), a tachykinin NK1-receptor antagonist; SR48968 (1 mg/kg), a tackykinin NK2-receptor antagonist; and CGRP(8-37) (50 nmol/kg), a CGRP1-receptor antagonist, administered intravenously before the intranasal application of the neuropeptides, inhibited the responses induced by SP, NKA and CGRP, respectively. In the guinea pigs sensitized with dinitrophenyl-coupled Ascaris suum allergenic extract, the intranasal antigen challenge caused nasal obstruction. The response was biphasic and consisted of the early phase response (EPR) and the late phase response (LPR), which developed 30 min and 6 h, respectively, after the antigen challenge. Intravenous administration of LY303870 (1 mg/kg) before the antigen challenge inhibited the EPR, while those of SR48968 (1 mg/kg) and CGRP(8-37) (50 nmol/kg) inhibited the LPR. The present results suggest that neuropeptides are involved in the allergic nasal obstruction.

Submicron emulsions containing 2.0% w/v pilocarpine as pilocarpine HCl, soybean oil (10% w/v) and egg lecithin (1.2% w/v) were formulated. Emulsions at pH 5.0, 6.5 and 8.5 were applied to the rabbit's eye, and the reduction in pupil diameter was measured for 6 h. The miotic effect was compared with that obtained with aqueous solutions at the same pH. A prolonged miotic effect was observed when the submicron emulsion was used as a vehicle. After application of emulsions at pH 5.0, 6.5 or 8.5, the time when 20% reduction of pupil diameter was still observed was 3.9 +/- 1.1 h, 4.3 +/- 1.3 h and 5.3 +/- 0.8 h, respectively, while, after application of a solution, this parameter was shorter by 30-40%. AUC(0-6h) values were larger after application of the submicron emulsions in comparison to aqueous solutions; however, statistically significant differences were only observed for emulsions at pH 6.5. Although the bioavailability of the drug is pH dependent, emulsions at higher pH cannot be considered for clinical use because of pilocarpine degradation which occurs with a similar rate as in aqueous solutions. Introduction of pilocarpine into the oily phase in the form of pilocarpine base or its oleate did not improve either the physicochemical or the pharmacological properties of the formulations. Irrespective of the pH and chemical form of pilocarpine used for emulsion preparation, practically all drug was found in the aqueous phase of the emulsion; thus, partitioning to the oily phase was negligible.

In clinical practice, photophobia resulting from persistent mydriasis may be associated with dysfunction of ocular parasympathetic nerves or primary iris lesions. We encountered a 5-year-old Miniature Dachshund and a 7-year-old Shih Tzu with mydriasis, abnormal pupillary light reflexes, and photophobia. Except for sustained mydriasis and photophobia, no abnormalities were detected on general physical examination or ocular examination of either dog. We performed pharmacological examinations using 0.1% and 2% pilocarpine to evaluate and diagnose parasympathetic denervation of the affected pupillary sphincter muscles. On the basis of the results, we diagnosed a pupillary abnormality due to parasympathetic dysfunction and not to overt primary iris lesions. The test revealed that neuroanatomic localization of the lesion was postciliary ganglionic in the first dog.

Adrenomedullin (ADM) is a vasodilator produced by vascular endothelium and smooth muscle cells. Although plasma ADM levels are increased in patients with hypertension, heart failure, and myocardial infarction, little information exists regarding the microvascular response to ADM in the human heart. In the present study we tested the hypothesis that ADM produces coronary arteriolar dilation in humans and examined the mechanism of this dilation. Human coronary arterioles were dissected and cannulated with micropipettes. Internal diameter was measured by video microscopy. In vessels constricted with ACh, the diameter response to cumulative doses of ADM (10−12–10−7 M) was measured in the presence and absence of human ADM-(22–52), calcitonin gene-related peptide-(8–37), N ω-nitro-l-arginine methyl ester (l-NAME), indomethacin (Indo),1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one, SQ-22536, or KCl (60 mM). ADM dilated human coronary arterioles through specific ADM receptors (maximum dilation = 69 ± 11%).l-NAME or N-monomethyl-l-arginine attenuated dilation to ADM (for l-NAME, maximum dilation = 66 ± 7 vs. 41 ± 13%, P < 0.05). Thus the mechanism of ADM-induced dilation involves generation of nitric oxide. However, neither1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one, SQ-22536, nor Indo alone altered dilation to ADM. High concentrations of KCl blocked dilation to ADM. The magnitude of ADM dilation was reduced in subjects with hypertension. We propose that, in human coronary arterioles, ADM elicits vasodilation in part through production of nitric oxide and in part through activation of K+ channels, with little contribution from adenylyl cyclase. The former dilator mechanism is independent of the more traditional pathway involving activation of soluble guanylate cyclase.

Purpose: To investigate the effect of changes in retinal illumination on Frequency Doubling Technology (FDT). Methods: Five eyes, of 5 adults who were free from identifiable ocular pathology, were examined using the Snellen chart and the Pelli-Robson chart, conventional automated perimetry, and the full threshold N-30 program of FDT. Each test was performed with and without a 0.9, 1.5, or 2.4 log unit neutral-density (ND) filter placed before the eye. Furthermore, the influence of pupil diameter on FDT test results was compared after treatment with pilocarpine or cyclopentolate with the influence of ND filters. Results: All tests showed a decrease in sensitivity with decreasing retinal illumination. Frequency Doubling Technology showed an especially pronounced and significant decrease in sensitivity. The maximum mean threshold difference in FDT results with ND filter was 31.2 dB while that with the Humphrey Field Analyzer and the Pelli-Robson chart were 13.3 dB and 0.66 log contrast, respectively. The mydriatic state of the pupil increased the sensitivity of FDT and the miotic state decreased it to about the same extent as the the 0.9 ND filter. Conclusion: The change in retinal illumination has more impact on FDT than on spatial contrast tests and conventional automated perimetry. It is important to take this into account in evaluating FDT results.

It is unknown which neurotransmitter receptors are involved in the transfer of sensory information from the digestive tract to the brainstem. We examined the potential involvement of central glutamate, acetylcholine, and CGRP receptors in vagal pathways arising from gut chemo- and mechanosensitive afferents. Unitary recordings were made from 21 vagal preganglionic efferent neurones. Neuronal discharge showed either excitation or inhibition in response to oesophageal balloon distension (1-2 ml), gastric distension (40-60 ml in whole stomach or 20 ml in proximal stomach), cholecystokinin-8s (100 pmol close intra-arterially) and bradykinin (18 nmol close intra-arterially). Effects of glutamatergic non-NMDA, muscarinic M1, and CGRP1 receptor antagonism on efferent responses were investigated using CNQX (75-155 nmol i.c.v.), pirenzepine (2.5-5.0 micromol/kg i.v.), and hCGRP8-37 (3.2-6.4 nmol i.c.v.), respectively. CNQX, pirenzepine, and hCGRP8-37, respectively, altered efferent responses in 65%, 23% and 41% of neurones. When both CNQX and hCGRP8-37 were administered, a further 58% of responses were reduced. CNQX and hCGRP8-37 reduced a proportion of efferent responses to all stimuli whereas pirenzepine selectively reduced only efferent responses to gastric distension. We conclude that central CGRP1 and non-NMDA receptors are involved in mediating a range of upper gastrointestinal mechano- and chemo-sensitive afferent inputs onto vagal efferents. M1 receptors, on the other hand, are selectively involved in neurotransmission from gastric mechanoreceptors.

To determine the effect of representative members from six classes of calcium channel blockers on intraocular pressure in the primate eye. Other antiglaucoma medications were administered with verapamil to determine their combined effect on intraocular pressure.Six healthy cynomolgus monkeys were anesthetized, and baseline intraocular pressure was measured. Drug-containing solution (50 microL) was instilled in one eye and intraocular pressure was measured in both eyes 90 minutes later.All classes of calcium channel blockers significantly lowered intraocular pressure in the treated eye. The percent reduction in intraocular pressure compared with the baseline pressure was 10% for verapamil (P0.002), 18% for nifedipine (P0.001), 15% for diltiazem (P0.001), 17% for flunarizine (P0.001), 19% for prenylamine (P0.001), and 6% for perhexiline (P0.01). In the fellow eye, a significant reduction in intraocular pressure was also seen with all calcium channel blockers except perhexiline, which suggested a crossover effect. In contrast, neither vehicle treated nor contralateral eyes showed a lowering of intraocular pressure when tested under the same conditions. In the treated eye, 0.5% timolol (P0.01) and 0.05% clonidine (P0.02) combined with 0.25% verapamil each appeared to produce an additive effect, with a significantly greater pressure-lowering effect than either agent alone. In addition, 0.005% pilocarpine (P0.001) and 0.00125% demecarium (P0.01) combined with 0.25% verapamil each appeared to produce a synergistic effect, with a significantly greater reduction in intraocular pressure than both agents combined.Topical calcium channel blockers and combinations of verapamil with antiglaucoma medications may provide a useful alternative for reducing intraocular pressure in patients with ocular hypertension or primary open-angle glaucoma.

Submicron emulsions containing pilocarpine as ion-pair with mono-dodecylphosphoric acid were prepared. Physical stability of these preparations was confirmed during 4 months of storage at 4 degrees C. Approximately 50% of the drug was found in the aqueous phase of emulsion separated using an ultrafiltration technique, while the rest was present in the oily phase and interphase. The miotic effect observed in rabbits after application of the ion-pair in aqueous solution or in submicron emulsion was the same; indicating that the drug distribution into the oily phase of the colloidal vehicle does not improve ocular bioavailability.

The expression of three aquaporin (AQP)-type water channels has been reported in the lacrimal gland: AQP5 in the apical membranes of acinar and duct cells, AQP4 in the basolateral membranes of acinar cells, and AQP1 in microvascular endothelia. Recent experiments indicate that water movement through AQP5 in the salivary gland is important in saliva secretion. To investigate the role of aquaporins in lacrimal gland function, basal and pilocarpine-stimulated tear secretion was compared in wildtype mice and knockout mice lacking AQP1, AQP4 and AQP5, as well as AQP3, which was found here to be expressed in the basolateral membrane of acinar cells. Tear fluid was collected in anesthetized mice using microcapillary tubes before and at 4 min intervals after pilocarpine administration. Tear fluid volumes were (in microliter per 4 min, S.E.): 0.69 +/- 0.06 (wildtype mice), 0.70 +/- 0.07 (AQP1 -/-), 0.81 +/- 0.13 (AQP3 -/-), 0.62 +/- 0.14 (AQP4 -/-), and 0.78 +/- 0.09 (AQP5 -/-) (differences not significant). Chloride concentrations (average 155 +/- 13 mM) measured by a fluorescence assay were also not different in tear fluid collected from wildtype vs aquaporin null mice. These findings provide direct evidence against an essential role for aquaporins in lacrimal gland fluid secretion. The requirement for aquaporins in salivary but not lacrimal gland secretion, may involve the substantially slower fluid secretion rate across lacrimal gland acinar cells.

Glaucoma in childhood is a diverse, blinding group of conditions, which presents a major therapeutic challenge. Treatment is primarily surgical with medical treatments used as an adjunct. None of these drugs has been granted approval by the regulatory agencies for use in children, but they are used on a compassionate basis. Issues of efficacy and safety of these medications in children are discussed. beta-adrenoceptor blockers have been employed as first line pharmaceutical therapy for many years. Recently three new classes of drugs have been developed for use in glaucoma in adults. beta-blockers remain first line therapy if there are no contraindications such as asthma. Topical carbonic anhydrase inhibitors (CAI) appear to be less effective than beta-blockers, but seem safe systemically, although associated with local irritation. They are useful as an adjunct to beta-blockers or as first line therapy when beta-blockers are contraindicated. Prostaglandins have not proved as effective in childhood glaucoma as in adult glaucoma, although it works well in some patients with juvenile open angle glaucoma (JOAG) and others with aphakic glaucoma. alpha-adrenergic agonists, although effective at least in the short-term, have serious, potential systemic side effects, which demand close observation when used in neonates and young infants.

To investigate whether a history of recurrent urinary tract infection (UTI) and/or the presence of day-wetting/urge symptoms might influence the contractile responses to the cholinergic agonist carbachol or to the sensory neuropeptide neurokinin A (NKA) in the urinary bladder in children.Small detrusor strips were taken from the margin of the cystotomy incision of the bladder dome during surgery to correct vesico-ureteric reflux (VUR) in 62 children (aged 4 months to 12 years) or for unrelated bladder conditions in five controls (aged 3 months to 13 years). Concentration-response curves to carbachol and NKA were constructed using organ-bath techniques, and results compared for age, sex, weight of the detrusor strip, UTI history or day-wetting syndrome.The contractile responses to NKA were no different for any of the features investigated. The contractile response to carbachol and NKA in detrusor from control and VUR patients was not significantly different. The children with a history of UTI were significantly older than those without. The contractility in response to carbachol was greater in older girls (aged 4-12 years) than younger girls (4 years) and than in all boys (4 years and 4-12 years; ANOVA P = 0.013). The mean (SEM) maximum contractile response to carbachol in the group of 20 young children (4-30 months) with previous UTI was 3.0 (0.3) g, whereas the maximum response in the age-matched group of 11 without UTI was 1.8 (0.3) g (P = 0.046). There were no significant differences in maximum responses between those with day-wetting and those without (aged4 years), although there was a significant difference in pD2 value, at 6.19 (0.13) and 5.58 (0.14), respectively (P = 0.018).Carbachol produced a larger contractile response in detrusor from children with a history of UTI than from those without, indicating possible alterations in muscarinic receptor characteristics. An increased sensitivity to muscarinic stimulation in day-wetting children was also suggested, whereas NKA is unlikely to be involved in any of these pathophysiological conditions.

PURPOSE More than one million patients in the United States are treated for glaucoma, although little is known about the typical clinical characteristics of this group of patients and the type of therapy they receive. This study was conducted to describe the demographic and diagnostic characteristics of patients beginning long-term drug therapy for glaucoma. METHODS This cross-sectional study included 544 patients beginning topical glaucoma medication regimens who received care at a group model health-maintenance organization (HMO) located in central Massachusetts. The primary medical records of 544 patients beginning topical glaucoma medication between 1987 and 1990 were reviewed to ascertain the presence of three clinical findings: intraocular pressure (IOP) > or = 22 mmHg; optic disc changes including cup-to-disc ratio > or = 0.8, cup-to-disc asymmetry > or = 0.2, or morphologic disc changes consistent with glaucomatous optic neuropathy; and visual field defect consistent with glaucoma. RESULTS A majority of the 544 patients (86%) were diagnosed as having primary open-angle glaucoma (POAG) by their physicians. Almost half (44.7%) of these patients had only an elevation in IOP without other clinical findings, and 9% met none of the above criteria for glaucoma according to information in the medical record. CONCLUSION In this setting, most patients who were prescribed drug therapy for POAG were treated for an elevation in IOP alone in the absence of other ophthalmologic characteristics of glaucoma.

We report on a 9-year-old girl diagnosed with ophthalmoplegic migraines who had been previously diagnosed, at age 7 years, with typical migraines with aura. After resolution of the third ophthalmoplegic migraine attack, the only evident residual clinical sign was Adie's pupil. During 24-month follow-up, at age 11 years, a neurologic examination produced completely normal results. However, Adie's pupil persisted. Adie's tonic pupil can be associated with extraocular diseases, which were all excluded in this patient. The mechanisms underlying tonic pupil are not fully understood. This is the first report, to the best of our knowledge, of an ophthalmoplegic migraine followed by persistent Adie's pupil. Possible pathogenic mechanisms are discussed.

The aim of this study is to evaluate the ocular hypotensive efficacy of a topical carbonic anhydrase inhibitor (dorzolamide) in primary open-angle glaucoma patients, administered alone or in association with beta-blockers or with beta-blockers and miotics, in a one-year follow-up.

Background: In the present study we evaluated the influence of topical miotics on intraocular pressure and the blood-aqueous barrier after uncomplicated phacoemulsification and PC-IOL implantation. Patients and methods: Fifty-two eyes were randomized into 2 groups: with miotics (n = 28) and without miotics (n = 24). The IOP was measured before, 6 h, 1 and 2 days after surgery. Measurement of aqueous flare was performed before and on days 1 and 2 after surgery. Patients with glaucoma, PEX or previous intraocular surgery were excluded. Results: In the group without miotics the IOP was 17.9 mm Hg (± 3.34) 6 h postoperatively; in the second group it was 15.5 mm Hg (± 3.25); P = 0.04. On the first postoperative day the IOP measured in the group without miotics was 15.3 mm Hg (± 2.70) and with miotics 13.0 mm Hg (± 2.28); P = 0.007. On the second day in the group without miotics the IOP was 13.9 mm Hg (± 3.05) and with miotics 12.60 mm Hg (± 2.19); P = 0.53. The changes in aqueous flare on the first and second day after surgery showed no significant influence of miotics on the blood-aqueous barrier (P > 0.05). Conclusions: Immediate postoperative application of topical miotics led to a small yet significant reduction of the IOP during the first 24 h after surgery. Our data suggest that there is no need for pharmacological reduction of the IOP after uncomplicated cataract surgery.

Objective: The aim of the present study was to determine the role of adrenomedullin (AM) in cardiac fibroblasts. Methods: The production and secretion of AM were examined in cultured neonatal rat cardiac fibroblasts, and the effects of AM on proliferation and protein synthesis of these cells were assessed by [3H]thymidine and [3H]phenylalanine incorporation, respectively. Results: Cultured cardiac fibroblasts secreted AM into the medium time-dependently at a rate of 20.3±3.0 fmol/5×104 cells/48 h, mean±S.D. Northern blot analysis showed expression of preproAM mRNA of 1.6 kb in these cells. In addition, 10−6 mol/l of angiotensin II (Ang II) and endothelin-1 (ET-1) significantly increased the AM secretion by 55 and 48%, respectively. Synthetic AM significantly reduced 10−6 mol/l Ang II- or 10−7 mol/l ET-1-stimulated [3H]thymidine and [3H]phenylalanine incorporation in a dose-dependent manner, and these effects were attenuated by a calcitonin gene-related peptide (CGRP) type 1 receptor antagonist, CGRP(8-37). Synthetic AM also had a dose-dependent stimulatory effect on cAMP accumulation in these cells, which was significantly attenuated by CGRP(8-37). A cAMP analogue, 8-bromo-cAMP, mimicked the AM effects, inhibiting the Ang II-stimulated [3H]thymidine and [3H]phenylalanine incorporation. Blockage of the effect of endogenous AM by anti-AM monoclonal antibody not only significantly reduced the basal level of intracellular cAMP, but also enhanced the [3H]thymidine and [3H]phenylalanine incorporation into the cells. Conclusions: Cultured neonatal rat cardiac fibroblasts produce and secrete AM, and the secreted AM may inhibit proliferation and protein synthesis of these cells. AM may exert these inhibitory effects partly by elevating intracellular cAMP. It is suggested that AM has an important role in modulating the growth of cardiac fibroblasts in an autocrine or a paracrine manner.

Unilateral microinjection of calcitonin gene-related peptide (CGRP, 1.6 pmol; 0.2 microl) into the ventromedial hypothalamus (VMH) and dorsomedial hypothalamus (DMH) immediately increased oxygen consumption (VO2), heart rate (HR), colonic temperature (Tco), and temperature of interscapular brown adipose tissue (TIBAT) in urethane-anesthetized rats, whereas vehicle saline injection into the VMH and CGRP injection into other hypothalamic regions such as the preoptic area, lateral hypothalamic area, paraventricular nucleus, and bed nucleus of the stria terminalis had no effect. The effects of CGRP injection into the VMH were dose-dependent over the range of 0.016-1.6 pmol. CGRP administration to the lateral ventricle (LV) required 16-320 pmol to elicit similar degrees of responses that were observed after the injection into the VMH. The increase in TIBAT was always higher than that in Tco after CGRP injection. Injection of [Cys(ACM)2,7]hCGRPalpha, a selective CGRP2 receptor agonist, did not induce any thermogenic effects. Human CGRP8-37, a proposed CGRP1 receptor antagonist, by itself induced heat production responses with no signs of inhibition of CGRP-induced responses. Thus, the receptor subtype of the thermogenic effect of CGRP could not be determined by the available pharmacological tools. The present results show that centrally administrated CGRP induces heat production in the BAT specifically through the VMH or DMH.

In order to obtain a novel ocular formulation with a potential for prolonging pilocarpine activity, the drug (2.0%) was incorporated into a submicron emulsion containing soya-bean oil and lecithin as emulgator. The effect of drug incorporation into the emulsion on its physical stability and on the other hand, the potential of the vehicle to reduce drug degradation at pH higher than 5.0 was studied. The pH was adjusted to 6.5 or 5.0 and the physicochemical stability of the formulations was observed. The mean diameter of oily particles in the resulting emulsions measured by a laser diffractometer was 0.6-0.7 micron and this was larger than in a drug-free emulsion where a 0.33 micron value was measured. The formulations were physically stable for 6 months at 4 degrees C, but progressing chemical degradation of pilocarpine was noted at pH 6.5. At that pH nearly 8% of pilocarpine was degraded to isopilocarpine and pilocarpic acid, both in the emulsion and in the solution. Thus, it may be concluded that pilocarpine in submicron emulsion is not protected against degradation. The presence of pilocarpine changes the physical stability of the vehicle since the formulation was easily destabilized during autoclaving or at room temperature. In the presence of higher concentration of lecithin (2.4%) or co-emulgators (poloxamer 2.0% or Tween 80 0.5%) the mean droplet size in the emulsions was the same as in a drug-free system. However the emulsions containing poloxamer were not stable during storage. Viscosity of pilocarpine emulsions can be increased by addition of methylcellulose or sodium carmellose (1.0%), but an intensive creaming occurs in these systems. Pilocarpine base is less suitable for emulsion preparation than hydrochloride salt, and emulsions prepared at pH 5.0 show the most satisfying stability.

AIM To determine the mechanical effects of pilocarpine on the trabecular-iris angle opening in eyes with narrow angles, compared with its effects on healthy control subjects with wide angles. METHODS A narrow angle was defined as 25° or less of trabecular-iris angle on ultrasound biomicroscopic examination. The change in anterior chamber depth (ACD), trabecular-iris angle (TIA), angle opening distance (AOD, distance between trabecular meshwork and iris) measured at 250 μm and 500 μm from the scleral spur (AOD250 and AOD500), and iris thickness was determined in 30 eyes of 30 patients (13 men and 17 women, between 63 and 82 years (mean 70.4 years)) with narrow angles and in 30 sex and age matched control subjects with wide angles before and 1 hour after the instillation of 2% pilocarpine hydrochloride by ultrasound biomicroscopy. RESULTS In all eyes with narrow angles, pilocarpine increased the TIA, AOD250, and AOD500; these changes increased significantly and linearly as the corresponding pretreatment values decreased ( r = 0.807, p = 0.0001; r = 0.787, p = 0.0001; r = 0.852, p = 0.0001). Of 30 eyes with wide angles, 23 eyes whose ACD was 2670 μm and more showed a decrease in the TIA, AOD250, and AOD500; the changes in TIA, AOD250, and AOD500 also significantly correlated with the corresponding pretreatment values ( r = 0.913, p = 0.0001; r = 0.882, p = 0.0001; r = 0.895, p = 0.0001). Pilocarpine induced a smaller decrease in ACD in eyes with narrow angles than in those with wide angles (p = 0.0001). There was a linear correlation between the increase in ACD change and the decrease in pretreatment ACD in eyes with narrow angles and those with wide angles ( r = 0.781, p = 0.0003; r = 0.798, p = 0.0001). CONCLUSIONS The finding that pilocarpine increases angular width in patients with narrow angles indicates that this agent is useful for treating patients with narrow angles and angle closure glaucoma. The prediction of the pilocarpine induced change in the angle may assist ophthalmologists in treating such patients.

Amylin (AMY) is a peptide of pancreatic origin principally involved in the carbohydrate metabolism, but that may interfere with central and peripheral dopamine (DA) pathways. The peptide, injected intracerebroventricularly (ICV) at the dose of 2.5 microg/rat, induced a decrease of copulatory activity in good copulators (GCO) and sluggish (SLU) male rats. The dose of 0.1 microg/rat did not affect significantly the sexual behavior of GCO rats, whereas AMY 0.5 microg/rat increased only the latency and reduced the frequency of ejaculation. At the dose of 2.5 microg/rat AMY antagonized the activation of sexual behavior induced by the DA receptor agonist, apomorphine administered subcutaneously (SC) at the dose of 100 microg/kg. Moreover, this inhibitory effect was blocked by the calcitonin gene-related peptide and AMY receptor antagonist, CGRP (8-37) fragment (injected ICV at the dose of 1 microg/rat). These data suggest that AMY may exert inhibitory effects on male sexual behavior in rats, probably interfering with central DA neurotransmission and with CGRP receptors.

Marfan syndrome is an autosomal dominant connective tissue disorder characterized by skeletal, cardiovascular, and ocular anomalies. Ectopia lentis is the most common ocular manifestation. We report an ocular sign not previously described in Marfan syndrome, iridocorneal adhesions secondary to anterior lens subluxation. Three patients with the Marfan syndrome had iridocorneal adhesions on slit-lamp examination. One patient developed adhesions following treatment with pilocarpine. She underwent pars plana vitrectomy and lensectomy in both eyes due to progression of the iridocorneal adhesions. Treatment with miotics rotates the lens-iris diaphragm anteriorly and may contribute to the formation of such adhesions. The two other patients remained stable and did not received lensectomy or vitrectomy. Careful slit-lamp examination of the anterior segment should be conducted in patients with dislocated lenses.