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Abstract Background Although the painful shoulder is one of the most common dysfunctions of the locomotor apparatus, and is frequently treated both at primary healthcare centres and by specialists, little evidence has been reported to support or refute the effectiveness of the treatments most commonly applied. According to the bibliography reviewed, physiotherapy, which is the most common action taken to alleviate this problem, has not yet been proven to be effective, because of the small size of sample groups and the lack of methodological rigor in the papers published on the subject. No reviews have been made to assess the effectiveness of acupuncture in treating this complaint, but in recent years controlled randomised studies have been made and these demonstrate an increasing use of acupuncture to treat pathologies of the soft tissues of the shoulder. In this study, we seek to evaluate the effectiveness of physiotherapy applied jointly with acupuncture, compared with physiotherapy applied with a TENS-placebo, in the treatment of painful shoulder caused by subacromial syndrome (rotator cuff tendinitis and subacromial bursitis). Methods/design Randomised controlled multicentre study with blind evaluation by an independent observer and blind, independent analysis. A study will be made of 465 patients referred to the rehabilitation services at participating healthcare centres, belonging to the regional public health systems of Andalusia and Murcia, these patients presenting symptoms of painful shoulder and a diagnosis of subacromial syndrome (rotator cuff tendinitis and subacromial bursitis). The patients will be randomised into two groups: 1) experimental (acupuncture + physiotherapy); 2) control (TENS-placebo + physiotherapy); the administration of rescue medication will also be allowed. The treatment period will have a duration of three weeks. The main result variable will be the change produced on Constant's Shoulder Function Assessment (SFA) Scale; as secondary variables, we will record the changes in diurnal pain intensity on a visual analogue scale (VAS), nocturnal pain intensity on the VAS, doses of non-steroid anti-inflammatory drugs (NSAIDs) taken during the study period, credibility scale for the treatment, degree of improvement perceived by the patient and degree of improvement perceived by the evaluator. A follow up examination will be made at 3, 6 and 12 months after the study period has ended. Two types of population will be considered for analysis: per protocol and per intention to treat. Discussion The discussion will take into account the limitations of the study, together with considerations such as the choice of a simple, safe method to treat this shoulder complaint, the choice of the control group, and the blinding of the patients, evaluators and those responsible for carrying out the final analysis.

The effects of the antihypertensive agent hydralazine (1 to 100 nmol/L) on the exocytotic process of single adrenal chromaffin cells have been studied using amperometry. Hydralazine does not reduce the frequency of exocytotic spikes but rapidly slows the rate of catecholamine release from individual exocytotic events by reducing the quantal size of catecholamine exocytosis. Confocal and standard epifluorescence microscopy studies show that hydralazine rapidly accumulates within secretory vesicles. The blockade of the vesicular H+ pump with bafilomycin A1 inhibits hydralazine uptake. Experiments with permeabilized cells show that hydralazine displaces catecholamines from secretory vesicles. The drug also displaces vesicular Ca2+, as shown by fura-2 microfluorimetry. These data suggest that hydralazine acts, at least partially, by interfering with the storage of catecholamines. These effects of hydralazine occurred within seconds, and at the tissue concentrations presumably reached in antihypertensive therapy; these concentrations are a thousand times lower than those described for relaxing vascular tissues in vitro. We proposed that these novel effects could explain many of the therapeutic and side effects of this drug that are likely exerted in sympathetic nerve terminals.

Amperometry is a powerful tool for studying the exocytotic process. Catecholamines released from a single secretory vesicle are oxidized by a carbon fiber microelectrode placed on a chromaffin cell membrane. This phenomenon can be recorded online as an amperometric signal. Each event of exocytosis is called a secretory spike. Several kinetic parameters can be extracted from spikes to get important information about catecholamine storage and to follow the time course of exocytosis. The large amount of data requires the use of computer programs. We describe software, written for Igor Pro (Wavemetrics, Lake Oswego, OR, USA), that allows the offline analysis of amperometric signals. It includes (1) the automatic analysis of a large collection of experiments without user intervention; (2) the visual check of located spikes; (3) data pooling from several experiments to create galleries with hundreds of thousands of spikes. In addition, we have designed a new filtering method for amperometric data. It provides an excellent tool to enhance the signal/noise ratio with minimal artifacts. This filter allows one to obtain more reliable spike parameters.

Amperometry is a widely used technique for monitoring the secretion of catecholamines (CA) by exocytosis. The use of carbon fibre microelectrodes allows the on-line recording of CA released from a single secretory vesicle. Amperometric signals are generated by oxidation of the quantally released CA close to the electrode tip. Each event of exocytosis is called a secretory spike. Here we describe a program written for IGOR (Wavemetrics, Lake Oswego, OR, USA), which may be used to analyze amperometric signals off-line. The procedures allow, (i) digital filtering and analysis of the current noise, spike identification and calculation of spike kinetic parameters; (ii) spike review; (iii) pooling spikes and data to create galleries, tables and histograms of measured parameters which can be exported to a graphic format or files for further analysis.

The effects of nitric oxide (NO) on the late phase of exocytosis have been studied, by amperometry, on Ba(2+)-stimulated chromaffin cells. Acute incubation with NO or NO donors (sodium nitroprusside, spermine-NO, S-nitrosoglutathione) produced a drastic slowdown of the granule emptying. Conversely, cell treatment with N(omega)-nitro-l-arginine methyl ester (a NO synthase inhibitor) or with NO scavengers (methylene blue, 2-(4-carboxyphenyl)-4,4,5, 5-tetramethyl-imidazoline-1-oxyl-3-oxide potassium) accelerated the extrusion of catecholamines from chromaffin granules, suggesting the presence of a NO modulatory tone. The incubation with phosphodiesterase inhibitors (3-isobutyl-1-methylxanthine or zaprinast) or with the cell-permeant cGMP analog 8-bromo-cGMP, mimicked the effects of NO, suggesting the involvement of the guanylate cyclase cascade. NO effects were not related to changes in intracellular Ba(2+). NO did not modify the duration of feet. Effects were evident even on pre-fusioned granules, observed under hypertonic conditions, suggesting that the fusion pore is not the target for NO, which probably acts by modifying the affinity of catecholamines for the intragranular matrix. NO could modify the synaptic transmitter efficacy through a novel mechanism, which involves the regulation of the emptying of secretory vesicles.

Exocytosis is the most common way for cells to secrete substances. This is a wide distributed phenomenon that involves all cell types and all animal species from yeast to human. Catecholamine (CA) release from adrenal chromaffin cells occurs by the exocytosis of secretory vesicles also called chromaffin granules. Individual secretory events can be easily monitored by the use of carbon fibers encased in glass microelectrodes. Catecholamines oxidized at the electrode tip produce a transient electrical current wave called secretory spike. The typical secretory spike consists in a sharp elevation of current followed by a rapid exponential decrease to the basal level. A 35-50% of the spikes showed a pre-spike wave called "foot" which represents the CA release through the fusion pore. The time course of secretory spikes observed is altered by extracellular phenomena's like diffusion. It has been suggested that amperometric spikes can be described by the convolution of a Gaussian with a decreasing exponential. The exponential function is partially governed by the diffusion of the secreted substances towards the electrode. In this article a method to deconvolve both functions is proposed. This mathematical approach allows the observation of the original secretory profile--the Gaussian--without the distortion caused by the diffusional broadening of catecholamines.