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Anti‐Wrinkle Therapy: Significant New Findings in the Non‐Invasive Cosmetic Treatment of Skin Wrinkles with Beta‐Glucan

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  • Prince Edward Island BioAlliance

Abstract and Figures

Oat beta-glucan is a water soluble, linear polymer of glucose consisting of 1,4 (70%) and 1,3 (30%) linkages with an average molecular weight of 1 × 106 Da. Scientific reports indicate beta-glucan is a film-forming moisturizer, a biological response modifier, and a promoter of wound healing. Our objective was to study the penetration of oat (1,4:1,3) beta-glucan in human skin models and to evaluate clinically its efficacy for reducing fine-lines and wrinkles. Penetration studies performed on human abdominal skin used a single application of 0.5% beta-glucan solution at a dose of 5 mg per cm2. The results showed that beta-glucan, despite its large molecular size, deeply penetrated the skin into the epidermis and dermis. A clinical study of 27 subjects was performed to evaluate the effects of beta-glucan on facial fine-lines and wrinkles. After 8 weeks of treatment, digital image analysis of silicone replicas indicated a significant reduction of wrinkle depth and height, and overall roughness. This work is the first ex vivo and in vivo demonstration of the physiological effects of beta-glucan in the penetration and restructuring of human tissue. The study supports the use of oat beta-glucan in the care and maintenance of healthy skin and the cosmetic treatment of the signs of aging.
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Reprint
VOLUME 8, NUMBER 1 JANUARY / MARCH 2005
magazine
The Global Publication of the International Federation of Societies of Cosmetic Chemists
R. Pillai, M. Redmond, J. Röding
Anti-Wrinkle Therapy: Significant New Findings in the
Non-Invasive Cosmetic Treatment of Skin Wrinkles with
Beta-Glucan
2
IFSCC Magazine – Reprint
INTRODUCTION
Oat has a long history of safe use to pro-
vide fast, temporary relief of the itching,
redness, and pain associated with many
minor skin irritations such as poison ivy/
oak/sumac, insect bites, and allergy [1]. In
the cosmetic application of beta-glucan,
consumers have described various bene-
fits including excellent, sustained mois-
turization properties together with an im-
proved, smoother appearance of the skin.
In recent years new wound product appli-
cations for beta-glucan have been found
in the management of partial thickness
burns, shallow abrasions, and laser treat-
ment [2, 3]. It has been reported that topi-
cal glucan administration enhances
wound healing by increasing macrophage
infiltration into the wound milieu, stimulat-
ing tissue granulation, collagen deposi-
tion, and re-epithelialization, together
with increasing the tensile strength of the
recovered wound [4, 5].
Laboratory experiments using beta-glu-
can from cereal (1,4; 1,3 linear glucose
polymer) and fungi (1,3; 1,6 branched glu-
cose polymer) indicated that all beta-glu-
cans are biological response modifiers,
with oat beta-glucan producing the great-
est cytokine induction activity in macro-
phages [6, 7].
The mechanism by which glucan or glu-
can-induced immunomodulators enhance
wound repair has remained elusive. We
do know that beta-glucan receptors exist
on mammalian macrophages and fibrob-
lasts [4, 8]; the effect of glucan on wound
repair is speculated to involve macro-
phage release of wound growth factors
with the further direct and indirect modu-
lation of fibroblast activity, including col-
lagen biosynthesis.
In the case of wounds, the disruption of
the dermal barrier gives clear and open
access of macrophages and fibroblasts to
topically applied beta-glucan. It remained
to be determined if cellular effects could
be achieved through normal, intact skin
and if the structure of aged skin could be
affected positively through the cosmetic
application of beta-glucan.
EXPERIMENTAL
Oat (1,4; 1,3) beta-glucan
Beta-glucan is the soluble fiber found in
the cell walls of oat kernels
(Figure 1)
.
Structurally, oat beta-glucan is a linear
polymer of glucose consisting of 1,4 (70%)
and 1,3 (30%) glycosidic linkages
(Figure
2)
.
The beta-glucan used in the present stud-
ies was extracted from oat and supplied
as a clear, viscous, 1% solution (Symrise
Inc., New Jersey). Briefly, the extraction
method comprised the aqueous, mild al-
kali (pH 9.2) extraction of beta-glucan from
oat bran, followed by the removal of pro-
tein, and ultrafiltration through a 0.1 mi-
Anti-Wrinkle Therapy: Significant New Findings
in the Non-Invasive Cosmetic Treatment of
Skin Wrinkles with Beta-Glucan
Ravi Pillai1, Mark Redmond2, Joachim Röding3
1Symrise Inc., 10 Gordon Drive, Totowa, New Jersey, USA
2Ceapro Inc. 1008 RTF University of Alberta, Edmonton, Alberta, Canada
3Symrise GmbH & Co KG., Bleichenbrücke 10, 20354 Hamburg, Germany
Corresponding author – email: ravi.pillai@symrise.com
Abstract
Oat beta-glucan is a water soluble, linear polymer of glucose consisting of 1,4 (70%) and 1,3 (30%) linkages with an
average molecular weight of 1x106Da. Scientific reports indicate beta-glucan is a film-forming moisturizer, a biolog-
ical response modifier, and a promoter of wound healing. Our objective was to study the penetration of oat (1,4 :1,3)
beta-glucan in human skin models and to evaluate clinically its efficacy for reducing fine-lines and wrinkles.
Penetration studies performed on human abdominal skin used a single application of 0.5% beta-glucan solution at a
dose of 5 mg per cm2. The results showed that beta-glucan, despite its large molecular size, deeply penetrated the
skin into the epidermis and dermis.
A clinical study of 27 subjects was performed to evaluate the effects of beta-glucan on facial fine-lines and wrinkles.
After 8 weeks of treatment, digital image analysis of silicone replicas indicated a significant reduction of wrinkle depth
and height, and overall roughness.
This work is the first
ex vivo
and
in vivo
demonstration of the physiological effects of beta-glucan in the penetration
and restructuring of human tissue. The study supports the use of oat beta-glucan in the care and maintenance of
healthy skin and the cosmetic treatment of the signs of aging.
Keywords: Beta-glucan, skin penetration, wrinkles, anti-aging
3
IFSCC Magazine – Reprint
cron filtration system. The resulting solu-
tion was double precipitated with ethanol
and resuspended to a final concentration
of 1%. The ultrafiltration of the beta-glu-
can solution produced a clear solution
confirmed by a low turbidity (<40 Neph-
elometric Turbidity Units: NTU). The mole-
cular weight range of the beta-glucan was
determined to be 0.5 x 106– 1.0x 106Da as
measured by the method of Wood [9].
Skin penetration study
In the first study we examined the dermal
penetration of (1,4; 1,3) beta-glucan into
sections of surgically-removed, human
abdominal skin. The penetration of beta-
glucan was visualized using Calcofluor
White, a beta-glucan specific fluorescent
stain. The use of Calcofluor White also
allowed semi-quantitative measurement
of beta-glucan penetration with fluores-
cence densitometry [10, 11].
Sections of abdominal tissue were re-
moved surgically without the
subcutaneous fat. The skin was
sliced to fit a penetration and
deposition chamber based
on a Franz Diffusion Cell. The
skin sections were first deep
frozen by liquid nitrogen and
sterilized by gamma-radiation,
which destroyed all yeast and
fungal elements that could in-
terfere with the assay. After ir-
radiation, the skin-section was
thawed and the specimens
were inspected for integrity be-
fore use with a pressure test.
Next, the skin section was con-
ditioned with respect to surface
temperature and moisture con-
tent. This condition was achiev-
ed by pre-heating the liquid
medium in the test chamber and
adjusting the air flow through the cham-
ber’s ventilation channel. A macroscopic
and physical examination of the skin spec-
imen was carried out before the test to
ensure suitability, and the area of the test
application site was 10cm2for all sam-
ples. During testing, the skin specimen
was supplied with a uniformly circulated
nutrient medium, which rinsed its lower
surface. The experimental conditions
were non-occlusive.
The test procedure involved one applica-
tion of 0.5% (w/w) beta-glucan solution
using a micro dose applicator at a dose of
5 mg per cm2of skin. After 8 hours of in-
cubation, the skin tissue was deep frozen.
It was then cut into thin slices and air
dried. Then the skin was cut from lower to
higher possible concentration, meaning
deeper dermis to horny layer.
The specimens were placed on thin glass
slides and allowed to dry. One drop of
Calcofluor White (BactidropTM, Remel,
Lenexa, KS, USA) was added and stained
for 30 seconds. The excess stain was re-
moved by washing with deionized water.
The specimens were then examined using
a fluorescent microscope with an excita-
tion wavelength ranging between 400
500 nm and a peak of 440 nm. Untreated
skin was used as the control.
The tests were done simultaneously with
two samples and one control for each vol-
unteer skin. All tests were repeated with
the skin of five volunteers.
Anti-aging study
In the second study, we performed a clin-
ical evaluation of the capacity of beta-
glucan to alleviate the extrinsic signs of
aging. The study was conducted in Col-
orado during the winter months to provide
a dry environmental challenge together
with a high exposure rate to UV.
The test was conducted on a panel of 27
subjects, with two carbomer gel formula-
tions; one contained 0.1% (w/w) (1,4; 1,3)
beta-glucan and the other was placebo.
The subjects applied the randomly as-
signed products twice daily, using a half-
face design. The subjects observed a
3-day conditioning period immediately
prior to baseline measurements. Each of
the 27 subjects treated their left and right
sides of the face, twice daily for eight
weeks. After 8 weeks of treatment, the
skin was evaluated for changes from
baseline values of various parameters
including fine lines, wrinkles and rough-
ness.
The clinical study included subjective and
objective assessments which were rec-
orded at baseline and 2, 4, and 8 weeks.
For the evaluation of fine-lines and wrin-
kles, silicone replicas of the outer canthus
of the eye area (crow’s feet) were sub-
jected to digital image analysis by expert
graders. Macrophotography was also
Figure 1: Fluorescent stained section of an oat
kernel. The beta-glucan present in the cell wall of
the oat fluoresces a brilliant blue when stained
with Calcofluor White.
Figure 2: Chemical structure of oat beta-glucan showing the beta 1,4 and beta 1,3 glycosidically linked glucose polymer structure.
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IFSCC Magazine – Reprint
used to evaluate the changes in fine lines
and wrinkles.
RESULTS AND DISCUSSION
The results of the skin penetration study
showed that (1,4; 1,3) beta-glucan had
penetrated the skin into the epidermis and
dermis
(Figures 3 and 5)
. No fluorescence
staining occurred in the control skin sec-
tion which was not treated with beta-glu-
can
(Figure 4)
. Quantitative assay of the
fluorimetric staining indicated that a sig-
nificant portion of the product (28.5% of
the applied beta-glucan) had entered the
skin
(Figure 6)
.
The clinical trial results indicated a higher
incidence of improvement with (1,4; 1,3)
beta-glucan than with the placebo.
Fig-
ure 7
shows the average percentage
change of the selected parameters from
the baseline, compared to the treatment
with placebo. The silicone replicas after
the test period demonstrated a smoothing
of the cutaneous surface after 8 weeks
of treatment with (1,4; 1,3) beta-glucan.
Macrophotography of the left and right
sides of the face also showed a reduction
in lines and wrinkles.
These results represent remarkable new
findings which will contribute to our un-
derstanding of the interaction of skin with
beta-glucan, and the ability of beta-glucan
to penetrate the skin deeply and elicit cel-
lular changes.
In the past, the potential ability of beta-
glucan to penetrate the skin was disre-
garded because it was thought that the
high molecular weight (> 0.5x106Da) of
the compound would prevent it from pen-
etrating into the epidermis and dermis,
and it would therefore be unable to inter-
Figure 3: Photograph of dermis skin sec-
tion treated with 0.05% (w/w) (1,4; 1,3) be-
ta-glucan solution (magnification x125).
Figure 4:Photograph of the control dermis
skin section (magnification x125).
Figure 5: High magnification photograph
of epidermis skin section treated with a
0.05% (w/w) solution of beta-glucan. Note
that beta-glucan staining is associated
with the inter-cellular matrix indicating
that the beta-glucan permeates the skin
by passing between cells rather than
passing through cells directly (magnifica-
tion x250).
stratum corneum epidermis dermis
Percentage of beta-glucan penetrated
Figure 6: Graphical analysis of the fluorimetric data obtained from the
skin penetration study. The blue bars represent the beta-glucan treat-
ed skin and the green bars represent the control skin. The results indi-
cate that beta-glucan is able to penetrate into the lower levels of the
skin, and therefore is able to interact with the fibroblasts and other
structural elements.
deep wrinkle all wrinkle average peak roughness
reduction reduction reduction reduction
Average percentage change from baseline
Figure 7: Graphical analysis of the facial results of the clinical trial obtained
through digital picture analysis after 8 weeks. The blue bars represent the
beta-glucan treated skin and the green bars represent the control skin.
5
IFSCC Magazine – Reprint
act with macrophages and fibroblasts.
Beta-glucan is able to adopt a number of
conformations and is typically extracted
in the form of aggregate particles > 1µm
which are clearly visible under a light
microscope. It is understandable that
such large particles may not be expected
to enter the skin and the effects of beta-
glucan were thought to be limited to the
skin’s surface.
However, the beta-glucan used in the
present penetration study was subject to
sub-micron filtration to produce an aggre-
gate-free, low-turbidity solution with no
particles visible under the light micro-
scope. Examination of the micrographs in
Figure 5
shows that the beta-glucan used
in our study does not enter the skin by
direct passage through the cells of the
epidermis and dermis, but instead works
its way into the skin by passing through the
inter-cellular matrix. Such a process may
be facilitated by a diffusion gradient and
by lipid and phospholipid interactions. In-
teractions of beta-glucan with lipids are
known and are the basis of the health-en-
hancing, lipid-controlling properties rec-
ognized by the FDA [12].
Having penetrated the skin to the dermis,
beta-glucan is able to interact with spe-
cific cells, namely macrophages and
fibroblasts. Results of
in vitro
experiments
have demonstrated that beta-glucan in-
teracts with macrophages to induce the
production of IL-1, which indirectly pro-
motes the production of procollagen by
fibroblasts. In addition, beta-glucan has
been shown to interact with fibroblast re-
ceptors, which results directly in the pro-
duction of procollagen [13, 14]. The con-
version of procollagen to collagen and its
incorporation into collagen bundles would
result in the type of effects noted in our
clinical study, specifically the facial skin
tightening leading to a reduction of fine
lines and wrinkles.
Questionnaires and subject follow-ups
indicated that the effects on fine lines
and wrinkles associated with use of beta-
glucan treatment were long-lived but not
permanent. With normal cellular turnover,
there was an appearance of fine lines. It
may be speculated that the continued use
of products containing beta-glucan would
result in a sustained improvement of ap-
pearance.
The results presented in the present study
offer a cosmetic alternative to other more
invasive treatments aimed at the reduc-
tion of fine lines and wrinkles in an aging
population. Injectable fillers like collagen
either from human, bovine, or porcine
sources are common, and recently
hyaluronic acid fillers have also been
introduced. Such fillers produce tempo-
rary, soft tissue with effects that last on
average for 3 to 4 months. With a similar
duration of effect, the cosmetic use of
Botulinum
toxin type A has been reported
to have increased multifold since 1997
[15]. Actives like retinoic acid and coen-
zyme Q10 are also used for the treatment
of wrinkles [16-18]. The regular and fre-
quent use of cosmetics containing oat (1,4;
1,3) beta-glucan is a new and exciting tool
in the fight against the signs of aging.
CONCLUSION
Oat (1,4; 1,3) beta-glucan is a natural
active ingredient offering significant per-
formance-enhancing properties for per-
sonal care applications. Our studies have
shown that the molecule, despite its con-
siderable molecular weight, is able to en-
ter the stratum corneum and epidermis
and penetrate deep into the dermis. The
observed effects of beta-glucan on tissue
restructuring and wrinkle reduction are
most likely effects mediated by fibroblast
stimulation and collagen deposition in the
dermis. These unique properties make oat
beta-glucan a promising and effective
ingredient for cosmetics.
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IFSCC Magazine – Reprint
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Apresenta-se neste eBook uma visão geral da produção cervejeira e dos insumos utilizados, contextualizando a etapa da mosturação, a qual é abordada de forma detalhada. A mosturação consiste numa das principais etapas da fase quente (inicial) da produção de cervejas, envolvendo a extração e transformação de moléculas presentes nos maltes e outros insumos cervejeiros para produção do mosto que será fermentado para produção da bebida. Durante a mosturação, de acordo com os procedimentos adotados, diversas enzimas poderão atuar, as quais apresentam substratos, temperaturas e pHs ótimos diferentes. Dessa forma, conhecer as enzimas presentes, como agem e suas consequências para produção é de extrema importância. Para tanto, apresentam-se as principais enzimas, relacionando-as com os possíveis resultados no mosto e na bebida final. Espera-se que a leitura deste eBook te auxilie na compreensão da importância e das ações das enzimas presentes na mosturação, auxiliando-o no planejamento consciente desta importante etapa envolvida na produção cervejeira.
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Striae gravidarum (SG) is a kind of dermal scar associated with psychosocial and therapeutic challenge. Topical reagents and non‐invasive laser are more preferred than invasive procedures for less pain and shorter downtime. However, there are few studies on comparing and combining these two modalities. The aim of the present study was to evaluate the efficacy and tolerance of a topical regimen containing β‐glucan, 1565‐nm non‐ablative fractional laser (NAFL; ResurFX), and combination of them for SG. A total of 128 unilateral abdomens from 64 subjects were randomly divided into four strategies and were followed up for 12 weeks: topical vehicle (Veh); topical β‐glucan (B); 1565‐nm NAFL combined topical vehicle (NAFL); 1565‐nm NAFL combined topical β‐glucan (B + NAFL). NAFL was applied three times with a 4‐week interval. Topical reagent was applied b.i.d. for 12 weeks. Global Aesthetic Improvement Scale (GAIS) scores were assessed by blinded physician according to standard photograph, and by subjects at 12th week. The degree of SG atrophy was assessed by blinded physician before treatment and at the 12th week according to a standard 5‐point scale. Collagen remodeling was evaluated by histological analysis and all adverse effects were recorded. A total of 56 women (112 unilateral abdomens) completed all study. The GAIS scores by blinded physician showed greater improvement in NAFL as compared with β‐glucan, and by subjects showed greater improvement in β‐glucan as compared with vehicle. In terms of striae atrophy scale, the improvement of SG atrophy was more prominent in NAFL compared to β‐glucan, and in β‐glucan + NAFL compared to Veh + NAFL. All treatments were well tolerated. Topical β‐glucan regimen can mildly improve SG. NAFL showed better results than topical β‐glucan regimen. The combined strategy may further improve the SG atrophy compared with single treatment strategy.
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Natural bioactive compounds (BCs) are types of chemicals found in plants and certain foods that promote good health, however they are sensitive to processing and environmental conditions. Microencapsulation by spray drying is a widely used and cost-effective approach to create a coating layer to surround and protect BCs and control their release, enabling the production of high functional products/ingredients with extended shelf life. In this process, wall materials determine protection efficiency, and physical properties, bioavailability, and storage stability of microencapsulated products. Therefore, an understanding of physicochemical properties of wall materials is essential for the successful and effective spray-dried microencapsulation process. Typically, polysaccharide-based wall materials are generated from more sustainable sources and have a wider range of physicochemical properties and applications compared to their protein-based counterparts. In this review, we highlight the essential physicochemical properties of polysaccharide-based wall materials for spray-dried microencapsulation of BCs including solubility, thermal stability, and emulsifying properties, rheological and film forming properties. We provide further insight into possibilities for the chemical structure modification of native wall materials and their controlled release behaviors. Finally, we summarize the most recent studies involving polysaccharide biopolymers as wall materials and/or emulsifiers in spray-dried microencapsulation of BCs.
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Previous studies have shown that beta-glucans extracted from yeast or fungi potentiate immune responses. In the present study, the immunomodulatory activities of beta-(1-->3,1-->4)-glucan, derived from oats, were investigated. The ability of oat beta-glucan (ObetaG) to stimulate IL-1 and TNF-alpha release from murine peritoneal macrophages and the murine macrophage cell line P338D1, was assessed. In vitro stimulation of macrophages with ObetaG resulted in the production of IL-1 in a dose and time-dependent manner, whereas only small amounts of TNF-alpha could be detected in the culture supernatants. ObetaG also induced the production of IL-2, IFN-gamma and IL-4 secretion in a dose-dependent manner in cultured spleen cells. The intraperitoneal administration of ObetaG in mice resulted in the accumulation of leucocytes, predominantly macrophages, in the peritoneal cavity. Furthermore, ObetaG was tested for its ability to enhance non-specific resistance to a bacterial challenge in mice. Survival of mice challenged with Staphylococcus aureus was enhanced by a single intraperitoneal administration of 500 microg of ObetaG 3 days prior to bacterial challenge. In conclusion, these studies demonstrated that ObetaG possesses immunomodulatory activities capable of stimulating immune functions both in vitro and in vivo.
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The processes of aging and photoaging are associated with an increase in cellular oxidation. This may be in part due to a decline in the levels of the endogenous cellular antioxidant coenzyme Q10 (ubiquinone, CoQ10). Therefore, we have investigated whether topical application of CoQ10 has the beneficial effect of preventing photoaging. We were able to demonstrate that CoQ10 penetrated into the viable layers of the epidermis and reduce the level of oxidation measured by weak photon emission. Furthermore, a reduction in wrinkle depth following CoQ10 application was also shown. CoQ10 was determined to be effective against UVA mediated oxidative stress in human keratinocytes in terms of thiol depletion, activation of specific phosphotyrosine kinases and prevention of oxidative DNA damage. CoQ10 was also able to significantly suppress the expression of collagenase in human dermal fibroblasts following UVA irradiation. These results indicate that CoQ10 has the efficacy to prevent many of the detrimental effects of photoaging.
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Glucans are (1-3)-beta-D-linked polymers of glucose that are produced as fungal cell wall constituents and are also released into the extracellular milieu. Glucans modulate immune function via macrophage participation. The first step in macrophage activation by (1-3)-beta-D-glucans is thought to be the binding of the polymer to specific macrophage receptors. We examined the binding/uptake of a variety of water soluble (1-3)-beta-D-glucans and control polymers with different physicochemical properties to investigate the relationship between polymer structure and receptor binding in the CR3- human promonocytic cell line, U937. We observed that the U937 receptors were specific for (1-->3)-beta-D-glucan binding, since mannan, dextran, or barley glucan did not bind. Scleroglucan exhibited the highest binding affinity with an IC(50)of 23 nM, three orders of magnitude greater than the other (1-->3)-beta-D-glucan polymers examined. The rank order competitive binding affinities for the glucan polymers were scleroglucan>schizophyllan > laminarin > glucan phosphate > glucan sulfate. Scleroglucan also exhibited a triple helical solution structure (nu = 1.82, beta = 0.8). There were two different binding/uptake sites on U937 cells. Glucan phosphate and schizophyllan interacted nonselectively with the two sites. Scleroglucan and glucan sulfate interacted preferentially with one site, while laminarin interacted preferentially with the other site. These data indicate that U937 cells have at least two non-CR3 receptor(s) which specifically interact with (1-->3)-beta-D-glucans and that the triple helical solution conformation, molecular weight and charge of the glucan polymer may be important determinants in receptor ligand interaction.
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The effect of intragastrically or parenterally administered β-glucan, extracted from oats, on the enhancement of disease resistance to Eimeria vermiformis was studied in mice. Groups of mice were immunosuppressed with dexamethasone (DXM), infected with oocysts of E. vermiformis and treated with oat β-glucan by the intragastric (i.g.) or subcutaneous (s.c.) routes. Faecal oocyst shedding was reduced in the β-glucan-treated groups compared to the non-treated group. Immunosuppressed mice which received no β-glucan treatment showed more severe clinical signs of the disease and a 50% mortality, while minimal clinical signs and no mortality were recorded in the β-glucan-treated groups. Total IgG, IgG1, IgG2a, IgM and IgA immunoglobulins in the serum of β-glucan-treated groups were overall higher than those in the non-treated group. Specific IgG anti-sporozoite and merezeite immunoglobulins in serum were significantly higher in the β-glucan-treated groups than in the non-treated aninals. No significant differences were found in the levels of intestinal IgA anti-sporozoite and anti-merozoite immunoglobulins. IFN-γ and IL-4-secreting cells, in response to sprozoite antigen, were detected in the spleen and mesenteric lymph nodes of the β-glucan-treated groups only. In conclusion, the i.g. and s.c. oat β-glucan treatment increased the resistance to E. vermiformis infection in immunosuppressed mice.
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Although topical applications of retinoids on rodents and humans have been shown to cause epidermal hyperplasia, a detailed study of the influence of retinoids on epidermal differentiation in vivo has not been performed. In order to assess the pharmacologic effects of chronic topical tretinoin application used to improve the appearance of patients with photoaged skin, cutaneous biopsies from 25 patients in a controlled clinical study were examined histologically and immunocytochemically. Chronic application of tretinoin causes epidermal thickening (25 of 25 samples), stratum granulosum thickening (15 of 25), parakeratosis (13 of 25), a marked increase in the number of cell layers expressing epidermal transglutaminase (13 of 25), and focal expression of two keratins, K6 (12 of 25) and K13 (8 of 25), not normally expressed in the epidermis. The morphologic changes correlated with immunohistochemical abnormalities; neither of these correlated with the subjective cosmetic response. Three major epidermal differentiation products, keratins K1, K10, and K14 were not altered, within the limits of the methods used. Thus, chronic topical tretinoin reprograms some, but not all, aspects of human epidermal differentiation in vivo.
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Immunomodulators that enhance macrophage function have been shown to be beneficial in a number of wound-healing models in humans and in experimental animals. The exact mechanism of this improved healing is unclear. To assess the role of collagen biosynthesis, the immunomodulator glucan phosphate was utilized in two murine models of wound healing, i.e., colon anastomosis and full-thickness skin incision. Tensile strength was evaluated using computer-assisted constant velocity tensiometry. Collagen biosynthesis was determined by assaying hydroxyproline content of wound hydrolysates by N-(9-fluorenyl)methoxycarbonyl/o-phthalaldehyde high-performance liquid chromatography. Experimental animals were treated with (1-3)-beta-D-glucan phosphate (250 mg/kg) intravenously 24 hours prior to colon anastomosis or skin incision. A second dose of glucan phosphate was given immediately postoperatively. Control animals received dextrose and water (5% w/v) intravenously. Tensile strength and hydroxyproline content were measured on postoperative Day 3. In the skin wound model, glucan phosphate treatment increased (P < 0.05) tensile strength by 42 per cent (342.5 +/- 12.2 vs 241.8 +/- 4.8 g), and hydroxyproline content was increased by 23.5 per cent (242.0 +/- 14.4 vs 196.8 +/- 10.5 pmol/microg; P < 0.05). In the glucan phosphate group, colon tensile strength was significantly (P < 0.05) increased by 34 per cent (34.2 +/- 2.3 g vs 45.8 +/- 2.1 g), and hydroxyproline content was increased by 7 per cent (47.45 +/- 3.31 vs 44.34 +/- 3.74 pmol/microg). These data indicate that macrophage modulation with glucan phosphate will increase tensile strength in experimental colon and skin wounds. In addition, we observed a positive correlation between glucan phosphate treatment, wound tensile strength, and collagen biosynthesis.
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The authors propose a treatment to improve skin texture and to decrease thin wrinkles and creases. The treatment is based on the use of 0.1% all-trans retinoic acid intradermic injections (with biopresence of 0.02%) combined with topic cream, immediately followed by 340 Nm blue light skin exposure. These procedures determine the retinoic binding protein stabilization that provides the acid intracellular penetration with its subsequent effects. An average of 10 sessions, once a week was required.
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We characterized the use of the fluorescent probe Sodium Green for measurements of intracellular free sodium using frequency-domain, phase-modulation fluorometry. The intensity decays were found to be strongly Na+ dependent, with mean lifetime increasing from 1.13 ns in the absence of Na+ to 2.39 ns in the presence of 140 mM Na+. Detailed analysis of the intensity decays in the presence of Na+ and K+ in the concentration range from 0 to 500 mM is provided. Sodium sensing using data measured at a single modulation frequency is described. Phase and modulation data showed high sensitivity to Na+ and substantially lower sensitivity to K+. Additionally, exposure of Sodium Green to intense illumination indicated that Sodium Green is much more photostable than its precursor, fluorescein. These results indicate that lifetime-based measurements with Sodium Green can be used for imaging of intracellular free [Na+] in the range from about 0.5 to 50 mM with high accuracy.