Principles for the safety evaluation of cosmetic powders.

Consumer exposure to cosmetic (personal care) products is mostly by dermal contact, however additional considerations with regards to potential inhalation exposure from some cosmetics, such as sprays and powders, may be needed for a robust and reliable safety assessment. To get a deeper understanding of the exposure to airborne particles and droplets during product application, a team of international experts was founded under the umbrella of the European Association of the Cosmetic Industry "Cosmetics Europe" (CE) in Brussels. This expert team has worked out a pragmatic strategy how small and medium sized enterprises (SMEs), but also relevant authorities, could handle the safety evaluation of cosmetic powder products. Sufficient information on the aerodynamic diameter of sprayed droplets and here specifically of airborne particles is essential in addition to knowing the exposure after typical product application. The current article is focused on the determination of inhalation exposure to solids, and the derivation of safe exposure levels for cosmetic powder products found in the market. The principles described herein are very similar to spray products as published earlier and should be applied in a similar way (Steiling et al., 2014). Prediction models for the best estimate of inhalation exposure, developed with data from computer simulation programs, individual real-time measurements or finally by experience from the market were introduced and applied. Safety assessment approaches for exposure from powder spray products were developed and have been already considered in regulatory guidelines like the EC Cosmetics Regulation.

Confirmation of in vitro and clinical safety assessment of behentrimonium chloride-containing leave-on body lotions using post-marketing adverse event data.

Behentrimonium chloride (BTC) is a straight-chain alkyltrimonium chloride compound commonly used as an antistatic, hair conditioning, emulsifier, or preservative agent in personal care products. Although the European Union recently restricted the use of alkyltrimonium chlorides and bromides as preservatives to ≤0.1%, these compounds have been safely used for many years at ≤5% in hundreds of cosmetic products for other uses than as a preservative. In vitro, clinical, and controlled consumer usage tests in barrier-impaired individuals were conducted to determine if whole body, leave-on skin care products containing 1-5% BTC cause dermal irritation or any other skin reaction with use. BTC-containing formulations were predicted to be non-irritants by the EpiDerm® skin irritation test and the bovine corneal opacity and permeability (BCOP)/chorioallantoic membrane vascular assay (CAMVA) ocular irritation test battery. No evidence of allergic contact dermatitis or cumulative dermal irritation was noted under the exaggerated conditions of human occlusive patch tests. No clinically assessed or self-reported adverse reactions were noted in adults or children with atopic, eczematous, and/or xerotic skin during two-week and four-week monitored home usage studies. These results were confirmed by post-marketing data for five body lotions, which showed only 0.69 undesirable effects (mostly skin irritation) reported per million shipped consumer units during 2006-2011; a value consistent with a non-irritating body lotion. No serious undesirable effects were reported during in-market use of the products. Therefore, if formulated in appropriate conditions at 1-5%, BTC will not cause dermal irritation or delayed contact sensitization when used in a whole-body, leave-on product.

A comparison of breast muscle characteristics in three broiler great-grandparent lines.

Genetic selection of broiler chickens has led to a gross overdevelopment of the broiler breast muscle pectoralis major. This may have resulted in increased myopathy and detrimental effects on meat quality. The present study examined 3 commercial great-grandparent lines (lines A, B, and C). Lines A and B are female lines, and line C is a male line. The mean BW of line C (2.7 kg) was significantly greater than those of lines A and B (both 2.3 kg). However, the mean breast yield of both lines B and C (8.9 and 8.7%, respectively) was significantly greater than that of line A (6.9%). Line B therefore matched the meat yield of line A while maintaining a high reproductive capacity. The mean breast fillet weight of line A (169 g) was significantly lower than lines B (207 g) and C (235 g). No differences were observed between lines in either mean fiber size or amount of connective tissue. Therefore, additional fibers must provide the additional weight in the breast fillet of lines B and C, compared with A. Plasma creatine kinase activity, a commonly used marker of muscle damage, was significantly higher in line A (1368 IU/L) than in lines B (995 IU/L) and C (982 IU/L). However, qualitative evaluations of muscle pathology revealed no differences among lines. Selection for increased embryonic muscle fiber number, rather than for increased radial fiber growth, could improve growth potential and may also alleviate muscle damage.

Skeletal muscle fibre growth and growth associated myopathy in the domestic chicken (Gallus domesticus).

1. Genetic selection of broilers may have pushed muscle fibres to their maximum functional size constraints. Broiler (B), female great-grandparent (GGP) and layer (L) lines were weighed, blood sampled and killed from 5 to 25 weeks of age. 2. At 25 weeks, Pectoralis major (Pm) fibre size reached by the B (65.9 microm) and GGP (59.8 microm) were 1.5 times greater than the L (38.1 microm). In the B and GGP lines, fibre growth of the Pm markedly exceeded that of the Biceps femoris (Bf) muscle. However, in the L line, fibre growth of the Pm and Bf muscle was comparable. Connective tissue content was generally higher in the Bf than in the Pm of all lines. 3. Centralised nuclei were observed predominantly in Pm, and may regulate fibre size. Both large muscle fibres and inadequate capillary supply may induce metabolic stress in B and GGP lines due to the large diffusion distances for oxygen, metabolites and waste products. 4. Enzyme markers of muscle damage (creatine kinase (CK), lactate dehydrogenase (LDH) and aspartate aminotransaminase (AST)) and histopathological analysis of Pm and Bf indicated greater myopathy in B and GGP vs L. 5. Regenerative processes were associated with oestrogen secretion. Reduced CK and LDH preceded egg yolk precursor production and increased calcium uptake for eggshell synthesis in all three lines. Oestrogen may stimulate muscle fibre regeneration and recovery as a myo-protective adaptation to potentially detrimental changes in calcium economy during egg production.

A stone farm: development of a method for simultaneous production of multiple calcium oxalate stones in vitro.

We have previously shown how individual calcium oxalate stones of about 1 cm can be grown in vitro. While this proved a design concept, it was severely limited as an experimental tool because of the time required to undertake comparative studies. Here we describe a development of this system in which six parallel pairs of stone generators are supplied with feed solutions generating a medium that is supersaturated with calcium oxalate. Twelve stones were grown simultaneously in aseptically prepared artificial urine over a period of 32 days from 100 mg to about 250 mg. Flow rates, pH and [Ca(2+)] were stable and reproducible over the course of the experiment. Sodium azide (0.02%) was included in the growth medium of six stones and caused a modest decrease in growth rate from 5.5 to 3.4 mg/day. The experimental design is such that this was readily detectable both visually and statistically ( p<0.001). This multiple stone growing system ("a stone farm") shows improved consistency and illustrates the statistical power of the technique. Azide has only a minor effect on the growth kinetics and can be used as an antibacterial agent in studies involving urinary macromolecules. The technique is suitable for practical and meaningful investigation of calcium oxalate stone formation in vitro.

The legal advantage of disability management.

Actively working with disabled workers to try to accommodate their needs and limitations--even if accommodation proves impossible--provides powerful protection from ADA lawsuits.

Histological and histochemical study of the vesicoureteric junction in infancy and childhood.

The morphology of the vesicoureteric junction was compared using 19 post mortem specimens obtained from male and female Afro-American and Caucasian children with an age range of 1 to 72 months (mean 4). All specimens were serially sectioned and the tissues processed using standard histological and histochemical techniques (acetylcholinesterase ¿AChE¿ and pseudocholinesterase ¿PChE¿). The results failed to reveal any differences in the structure of the vesicoureteric junction with respect to age, sex and ethnic origins. The vesicoureteric junction comprised 3 histologically and histochemically distinct smooth muscle components. Ureteric muscle formed a complete inner layer rich in PChE which continued beyond the ureteric orifices to merge distally with the superficial trigone. An intermediate layer of muscle was also demonstrated whose constituent muscle cells possessed specific histological features and which was rich in both AChE and PChE, which is distinct from that derived from the ureter and detrusor. The presence of detrusor muscle on the outer aspect of the juxtavesical segment of ureter rich in AChE was also confirmed. While this study, using histochemical studies in infants and children, did not reveal any differences in the structure of the vesicoureteric junction with respect to age, sex or ethnic origin, an intermediate layer of muscle was identified with histochemical characteristics more like that of the male genital tract than that derived from the ureter or detrusor muscle.

Increase in presumptive sensory nerves of the urinary bladder in idiopathic detrusor instability.

The density of subepithelial, presumptive sensory nerves in the bladder wall was assessed in 21 women with idiopathic detrusor instability and compared with the density of these nerves in 21 asymptomatic women, using a point-counting technique on sections of bladder biopsies stained for acetylcholinesterase activity. The mean value (+/- S.E.) for the amount of such nerves in patients with detrusor instability (91 +/- 13/mm2) was significantly greater than that from the control group (61 +/- 7/mm2). This suggests that a relative abundance of subepithelial sensory nerves may serve to increase the appreciation of bladder filling, giving rise to the frequency and urgency of micturition which are characteristic of patients with detrusor instability.

The innervation of the human prostate gland--the changes associated with benign enlargement.

Different regions of the prostate gland, namely prostatic capsule, peripheral prostate and central prostate (subdivided into proximal (near the bladder neck), distal (near the verumontanum) and midway between these areas) were obtained from 32 obstructed (stable obstructed, n = 8; unstable obstructed, n = 13; acute retention, n = 11) and five control patients. The innervation of these tissues was studied both histochemically to localise acetylcholinesterase activity and immunohistochemically for dopamine-beta-hydroxylase, 5-hydroxytryptamine, vasoactive intestinal polypeptide, neuropeptide Y, leu- and met-enkephalin, calcitonin gene-related peptide, substance P and somatostatin. In control patients the greatest density of nerves was found in the proximal central prostate, followed by the anterior capsule and distal central prostate, with the least density in the peripheral prostate. The greatest density of nerves were acetylcholinesterase positive and immunoreactive to neuropeptide Y followed (in decreasing order) by nerves immunoreactive to: vasoactive intestinal polypeptide and dopamine beta-hydroxylase; leu-enkephalin and 5-hydroxytryptamine; calcitonin gene-related peptide; met-enkephalin; substance P; somatostatin. In addition a group of periacinar 5-hydroxytryptamine-immunoreactive cells and ganglia containing acetylcholinesterase, dopamine beta-hydroxylase and all of the peptides studied except somatostatin were identified. In the prostate gland from obstructed patients there was a significant reduction in the density of acetylcholinesterase-positive nerves (p less than 0.001) when compared with the controls. A similar trend was found for dopamine beta-hydroxylase, 5-hydroxytryptamine and all of the putative neuropeptides in most areas of the prostate, the most notable exceptions being in the peripheral prostate, with an increase in dopamine beta-hydroxylase- and leu-enkephalin-immunoreactive nerves in all three groups of obstructed patients an an increase in vasoactive intestinal polypeptide- and calcitonin gene-related peptide-immunoreactive nerves in those presenting in urinary retention. The functional significance of these findings is discussed.

Cystometric, physiological and morphological studies after relief of bladder outflow obstruction in the pig.

Experimental bladder outflow obstruction was relieved in 18 pigs between 2 and 15 months after the creation of partial urethral obstruction. Cystometric, physiological and morphological studies were performed 2 to 6 months after relief of the obstruction. An increase in average voiding flow rates from 2.8 +/- 1.0 ml/s to 6.8 +/- 1.2 ml/s was recorded in the Landrace pigs and from 2.2 +/- 0.9 ml/s to 7.4 +/- 1.4 ml/s in the Göttingen mini-pigs. There was a concomitant decrease in the voiding detrusor pressures from 52 +/- 11 cm H2O to 32 +/- 8 cm H2O and from 78 +/- 12 cm H2O to 33 +/- 6 cm H2O respectively. A return towards control values of the physiological responses to exogenously applied agonists (acetylcholine and potassium) and to electrical field stimulation was observed. There was an increase in neuronal innervation in the morphological studies which was more marked in the animals with a shorter period of obstruction. The implications for patient care are discussed.

Relationship between bladder morphology and long-term outcome of treatment in patients with high pressure chronic retention of urine.

A group of 32 men undergoing bladder outflow surgery for high pressure chronic retention (HPCR) of urine were studied prospectively. At the time of treatment marked morphological changes in the bladder wall were demonstrated histologically, but after a mean follow-up of 42.9 months residual urine had decreased significantly and renal function had improved or stabilised in 28 patients (84%). Four patients deteriorated but in 3 of these another potential cause for loss of renal function was present. The majority of patients have a good long-term prognosis following treatment for HPCR.

Sequential morphological changes in the pig detrusor in response to chronic partial urethral obstruction.

The sequential morphological changes which occur in the wall of the porcine urinary bladder in response to experimentally induced partial outflow obstruction have been determined using morphometric light and electron microscope techniques. The initial morphological response to urethral obstruction was a reduction of approximately 50% in the density of autonomic nerves in the bladder wall which occurred within the first 3 months. During this period the morphology of smooth muscle cells and the distribution of connective tissue were unaltered. Longer periods of obstruction produced a gradual further reduction in the density of innervation. However, from 3 months onwards the detrusor muscle bundles became infiltrated by connective tissue, although significant change in muscle cell size was not detected after obstruction for 12 months. Using electron microscopy, smooth muscle cells were observed to possess extensive amounts of perinuclear granular reticulum, this feature being particularly marked in bladders subjected to longer periods of obstruction. The underlying mechanisms responsible for these structural changes remain to be determined.

The pathogenesis of genitourinary prolapse and stress incontinence of urine. A histological and histochemical study.

Histological and histochemical analysis of biopsy samples of pubococcygeus muscle obtained from asymptomatic women and from women with stress incontinence of urine, with or without genitourinary prolapse, have been compared. In the asymptomatic women both age and parity appeared to be related to the morphological features of the samples and in particular those obtained from the posterior part of the pubococcygeus. In the symptomatic women there was a significant increase in the number of muscle fibres showing pathological damage which were obtained from the posterior part of the pelvic floor. The range of diameters of both Type I and Type II fibres obtained from this region was significantly different between symptomatic and asymptomatic women. These findings may be attributable to partial denervation of the pelvic floor in patients with urinary stress incontinence with or without genital tract prolapse.

Bladder outflow obstruction--a cause of denervation supersensitivity.

Eighteen Landrace pigs and 12 Göttingen mini-pigs were evaluated in a study of experimental bladder outflow obstruction. Twenty-two of the animals underwent partial bladder outflow obstruction for periods up to 12 months. The subsequent changes were assessed using cystometric, physiological and morphological means. There was a consistent increase in the voiding pressures and a concomitant reduction in the flow rates in all the obstructed animals. Seventy-seven per cent of the obstructed animals showed cystometric evidence of bladder instability. In vitro studies showed an increase in sensitivity to exogenously applied agonists and a reduction in sensitivity to intramural nerve stimulation. Morphological studies showed an inverse correlation between neuronal density and the duration of obstruction. These changes are typical of post-junctional supersensitivity secondary to partial denervation. These results suggest that agents capable of stabilising the bladder smooth muscle membrane may be useful in the treatment of detrusor instability secondary to bladder outflow obstruction.

Quantitative study of phenol as a neurolytic agent in the urinary bladder.

The endoscopic subtrigonal injection of a 6% aqueous phenol solution is an effective technique for denervating the bladder but its clinical usefulness is limited by unpredictable side effects. This study explored the possibility of making this procedure safer by comparing the neurolytic effects of different concentrations and carriers of phenol. Phenol in 2.5 and 5% solutions in three different carriers (water, glycerine and oil) was injected into the rectovesical pouch in 35 rats. After 3 weeks the bladders were excised and the effects on the density of acetylcholinesterase-positive nerves were assessed in each animal using morphometric techniques. The density of enzyme-containing nerves was reduced by 20% in the phenolised animals when compared with controls. This reduction was maximum when water was used as the carrier for either 2.5 or 5% phenol solutions.

The effects of vasectomy on the autonomic innervation of the human vas deferens.

Neurohistochemical and fine structural techniques have been employed to examine the intramural autonomic innervation of the human vas deferens following surgical division of the duct one to 15 years previously. Samples from sites on the distal (testicular) and proximal (urethral) aspects of the original vasectomy have been compared with control specimens obtained at vasectomy as to the arrangement and distribution of autonomic nerves. In contrast with tissue from the proximal part and from controls, the distal samples revealed a marked reduction in the noradrenergic innervation of the muscle coat. In addition acetylcholinesterase-containing nerves associated with the basal aspect of the epithelium were usually absent from the distal portion of the vas deferens. These findings have been considered in relation to the contractile and secretory activities of the organ following vasovasostomy and may be of importance to the maturation and fertility of spermatozoa.

PIP: Vas deferens specimens taken from 43 men during vasovasostomy were examined by histochemical techniques and electron microscopy for evidence of noradrenergic and acetylcholinergic nerve structures. The subjects had been vasectomized 1-15 years ago (mean 6.5 years). Controls were 22 men being operated for vasectomy. Light microscopy revealed fine noradrenergic nerve plexi throughout the muscle coat, in the proximal, urethral portions of the vas, and circling blood vessels in the adventitia, as well as in both proximal and distal control specimens. The distal, testicular specimens of previously vasectomized men contained only occasional adrenergic fibers. In proximal or urethral portions, fine cholinergic nerves occurred in the muscle coat; large ones in the adventitia near blood vessels, and numerous cholinergic fibers were located under the epithelium. In distal, testicular portions, cholinergic nerves were similar to controls and to urethral specimens from vasectomized men in the muscle layer only, but absent in lamina propria. Ultramicroscopic structure of these various nerve fibers is described in detail. It is to be noted that the distal portion of the vas is the testicular portion, as concerns innervation, and this is lost after vasectomy. It was remarkable to see evidence of reinnervation of cholinergic axons distally, however. It has been established that noradrenergic control is important for the motor and secretory activities of the vas. Physiological significance of adrenergic innervation is unknown. These results add evidence for the advice to men seeking vasectomy to consider it a permanent form of fertility control.

The effect of age on the autonomic innervation of the urinary bladder.

Quantitative methods have been used to assess the amount of autonomic nerve in bladder biopsy samples from a group of "control" patients ranging in age from 20 to 79 years. Each patient included in the study was urodynamically normal and showed no subjective evidence of bladder trabeculation at cystoscopy. Using light microscopy, a significant linear reduction in the amount of acetylcholinesterase-positive nerve was observed with increasing age. Counts of axon profiles and measurement of smooth muscle cell cross-sectional areas in the electron microscope revealed a similar reduction in the amount of nerve per mm2 of detrusor muscle tissue. These findings thus confirm that a real reduction in the number of nerve axons occurs with age and the light microscope results are not due merely to a reduction in the amount of nerve-associated acetylcholinesterase. The present study provides baseline data for future comparisons of the density of autonomic innervation in bladders which are functionally impaired.

Decrease in the autonomic innervation of human detrusor muscle in outflow obstruction.

In a group of patients in whom bladder outflow obstruction had been confirmed urodynamically, quantitative assessment of the amount of autonomic nerve in detrusor biopsy samples has been carried out using light and electron microscope techniques. In each specimen allowance was made for muscle cell hypertrophy and increases in connective tissue, both of which occurred in response to bladder outflow obstruction. Similar quantitative assessment was performed on bladder biopsy samples from a group of unobstructed 'control' patients. When the results from the two groups were compared a statistically significant reduction in the amount of autonomic nerve supplying detrusor muscle was demonstrated in the obstructed group. This finding provides additional evidence that functional impairment of the urinary bladder occurs in response to outflow obstruction and emphasizes the need for prompt relief of the condition.

Bladder muscle biopsy and urethral sphincter EMG in patients with bladder dysfunction after pelvic surgery.

Eleven patients who suffered persistent bladder dysfunction after pelvic surgery have been investigated by needle urethral sphincter electromyography (EMG) and bladder muscle biopsy, and the results compared with those obtained in a series of controls. Individual motor units recorded from the urethral sphincter in patients who had undergone pelvic surgery were strikingly abnormal, suggesting the presence of reinnervation, and the density of detrusor innervation was significantly reduced. However, since reduction in the density of detrusor innervation may occur in circumstances other than peripheral nerve injury, we conclude that urethral sphincter EMG provides the most effective means of assessing damage to vesico-urethral innervation as a result of previous pelvic surgery.