LOW-INTENSITY PULSED ULTRASOUND PRODUCED AN INCREASE OF OSTEOGENIC GENES EXPRESSION DURING THE PROCESS OF BONE HEALING IN RATS

The aim of this study was to measure the temporal expression of osteogenic genes during the process of bone healing in low-intensity pulsed ultrasound (LIPUS) treated bone defects by means of histopathologic and real-time polymerase chain reaction (PCR) analysis. Animals were randomly distributed into two groups (n = 30): control group (bone defect without treatment) and LIPUS treated (bone defect treated with LIPUS). On days 7, 13 and 25 postinjury, 10 rats per group were sacrificed. Rats were treated with a 30 mW/cm(2) LIPUS. The results pointed out intense new bone formation surrounded by highly vascularized connective tissue presenting a slight osteogenic activity, with primary bone deposition was observed in the group exposed to LIPUS in the intermediary (13 days) and late stages of repair (25 days) in the treated animals. In addition, quantitative real-time polymerase chain reaction (RT-qPCR) showed an upregulation of bone morphogenetic protein 4 (BMP4), osteocalcin and Runx2 genes 7 days after the surgery. In the intermediary period, there was no increase in the expression. The expression of alkaline phosphatase, BMP4 and Runx2 was significantly increased at the last period. Our results indicate that LIPUS therapy improves bone repair in rats and upregulated osteogenic genes, mainly at the late stages of recovery. (E-mail: a.renno@unifesp.br) (C) 2010 Published by Elsevier Inc. on behalf of World Federation for Ultrasound in Medicine & Biology.

Low-intensity Ultrasound Increases FAK, ERK-1/2, and IRS-1 Expression of Intact Rat Bones in a Noncumulative Manner

Background Low-intensity pulsed ultrasound stimulation (LIPUS) reportedly increases osteogenesis in fracture models but fails in intact bone, suggesting LIPUS does not act on mechanotransduction and growth factor pathways of intact bone. Questions/Purposes We asked whether daily 20-minute LIPUS applied to intact tibias would act on bone proteins involved in mechanotransduction (focal adhesion kinase [FAK], and extracellular signal-regulated kinase-1/2 [ERK-1/2]), and growth factor signaling (insulin receptor substrate-1 [IRS-1]) pathways at 7, 14, and 21 days of treatment. Methods Immunoblotting was performed to detect FAK, ERK-1/2, and IRS-1 expression and activation from the stimulated intact tibias at 7, 14, and 21 days of daily 20-minute LIPUS. Results LIPUS increased FAK expression (at 7 days), ERK-1/2 (at 14 days), and IRS-1 (at 7 days), but expression decreased 7 days later, indicating a noncumulative effect of LIPUS. As only FAK expression was detected at 21 days, these observations suggest LIPUS influences nuclear reactions that may be modulated by a major cellular mechanism preferentially inhibiting IRS-1 expression and not FAK expression. Increased ERK-1/2 expression at 14 days suggests the differing mechanisms for promoting ERK-1/2, FAK, and IRS-1 syntheses. IRS-1 expression behaved similarly to FAK expression; therefore, LIPUS may modulate growth factor pathways. LIPUS increased sustained FAK and ERK-1/2 activation, but not IRS-1, suggesting sustained ERK-1/2 activation is not the result of mechanically induced growth factor activation. Conclusions LIPUS acts on mechanotransduction and growth factor pathways in intact bone in a noncumulative manner. Clinical relevance These data suggest LIPUS applied to intact bone acts on proteins involved in osteogenesis.

Is Nitric Oxide a Mediator of the Effects of Low-Intensity Electrical Stimulation on Bone in Ovariectomized Rats?

Low-intensity electrical stimulation (LIES) may counteract the effects of ovariectomy (OVX) on nitric oxide synthase (NOS) expression, osteocyte viability, bone structure, and microarchitecture in rats (Lirani-Galvo et al., Calcif Tissue Int 84:502-509, 2009). The aim of the present study was to investigate if these effects of LIES could be mediated by NO. We analyzed the effects of NO blockage (by l-NAME) in the response to LIES on osteocyte viability, bone structure, and microarchitecture in OVX rats. Sixty rats (200-220 g) were divided into six groups: sham, sham-l-NAME (6 mg/kg/day), OVX, OVX-l-NAME, OVX-LIES, and OVX-LIES-l-NAME. After 12 weeks, rats were killed and tibiae collected for histomorphometric analysis and immunohistochemical detection of endothelial NOS (eNOS), inducible NOS (iNOS), and osteocyte apoptosis (caspase-3 and TUNEL). In the presence of l-NAME, LIES did not counteract the OVX-induced effects on bone volume and trabecular number (as on OVX-LIES). l-NAME blocked the stimulatory effects of LIES on iNOS and eNOS expression of OVX rats. Both l-NAME and LIES decreased osteocyte apoptosis. Our results showed that in OVX rats l-NAME partially blocks the effects of LIES on bone structure, turnover, and expression of iNOS and eNOS, suggesting that NO may be a mediator of some positive effects of LIES on bone.

Low-Intensity Electrical Stimulation Counteracts the Effects of Ovariectomy on Bone Tissue of Rats: Effects on Bone Microarchitecture, Viability of Osteocytes, and Nitric Oxide Expression

Low Intensity Electrical Stimulation (LIES) has been used for bone repair, but little is known about its effects on bone after menopause. Osteocytes probably play a role in mediating this physical stimulus and they could act as transducers through the release of biochemical signals, such as nitric oxide (NO). The aim of the present study was to investigate the effects of LIES on bone structure and remodeling, NOS expression and osteocyte viability in ovariectomized (OVX) rats. Thirty rats (200-220 g) were divided into 3 groups: SHAM, OVX, and OVX subjected to LIES (OVX + LIES) for 12 weeks. Following the protocol, rats were sacrificed and tibias were collected for histomorphometric analysis and immunohistochemical detection of endothelial NO synthase (eNOS), inducible NOS (iNOS), and osteocyte apoptosis (caspase-3 and TUNEL). OVX rats showed significant (p < 0.05 vs. SHAM) decreased bone volume (10% vs. 25%) and trabecular number (1.7 vs. 3.9), and increased eroded surfaces (4.7% vs. 3.2%) and mineralization surfaces (15.9% vs. 7.7%). In contrast, after LIES, all these parameters were significantly different from OVX but not different from SHAM. eNOS and iNOS were similarly expressed in subperiosteal regions of tibiae cortices of SHAM, not expressed in OVX, and similarly expressed in OVX + LIES when compared to SHAM. In OVX, the percentage of apoptotic osteocytes (24%) was significantly increased when compared to SHAM (11%) and OVX + LIES (8%). Our results suggest that LIES counteracts some effects of OVX on bone tissue preserving bone structure and microarchitecture, iNOS and eNOS expression, and osteocyte viability.

Collaboration intensity in the Brazilian supermarket retail chain

Purpose - This paper seeks to identify collaboration elements and evaluate their intensity in the Brazilian supermarket retail chain, especially the manufacturer-retailer channel. Design/methodology/approach - A structured questionnaire was elaborated and applied to 125 representatives from suppliers of large supermarket chains. Statistical methods including multivariate analysis were employed. Variables were grouped and composed into five indicators (joint actions, information sharing, interpersonal integration, gains and cost sharing, and strategic integration) to assess the degree of collaboration. Findings - The analyses showed that the interviewees considered interpersonal integration to be of greater importance to collaboration intensity than the other integration factors, such as gain or cost sharing or even strategic integration. Research limitations/implications - The research was conducted solely from the point of view of the industries that supply the large retail networks. The interviews were not conducted in pairs; that is, there was no application of one questionnaire to the retail network and another to the partner industry. Practical implications - Companies should invest in conducting periodic meetings with their partners to increase collaboration intensity, and should carry out technical visits to learn about their partners` logistic reality and thus make better operational decisions. Originality/value - The paper reveals which indicators produce greater collaboration intensity, and thus those that are more relevant to more efficient logistics management.

Correlation Between Autofluorescence Intensity and Tumor Area in Mice Bearing Renal Cell Carcinoma

Protoporphyrin IX (PpIX) is a porphyrin derivative that is accumulated in cancerous tissue in consequence of the tumor-specific metabolic alterations. The aim of this study was to evaluate the accumulation of PpIX in mice bearing renal cell carcinoma by spectroscopy analysis. A total of 24 male Balb/c mice, 6 weeks old, were divided into six groups: Normal (without inoculation of tumor cells) and 4, 8, 13, 16, and 20 days after inoculation of tumor cells. The orthotopic tumor model of renal cancer was used. Murine renal cell carcinoma (Renca cells) were inoculated into the subcapsular space of the kidney. Normal and tumor-bearing kidneys in different progression stages were removed and analyzed by ex-vivo spectroscopy and by microscopy, for tumor histometric analysis. Emission spectra were obtained by exciting the samples at 405 nm. Significant differences between normal and tumor-bearing kidneys in autofluorescence shape occurred in the 600-700 nm spectral region. A good correlation was found between emission band intensity at 635 nm and the tumor area.

South American leaf blight intensity evaluated in six clones of young and adult rubber trees in the Vale do Ribeira region, Sao Paulo state, Brazil

Rubber tree clones present different intensity of symptoms, depending on their age. This is mostly clearly seen in the presence or absence of old leaves with ascopores, corresponding to flows of new and susceptible leaves. The objective of this work was to evaluate the intensity of symptoms of south American leaf blight (SALB) in six rubber tree clones, one and eight years old, in the Vale do Ribeira region, Sao Paulo state. The results Showed that clones FX 3864, RRIM 600, IAN 873 and IAN 717 Suffered more attacks when young, and RRIM 600, IAN 717 and FX 3864 when adult. The clone IAN 873 showed the smallest amounts of disease in the adult phase, due to uniform change in the leaves and a compact flow of new leaves, which happened during a season that was unfavorable to pathogen infection, presenting the phenomenon of avoidance or evasion in time.

The potential for increasing rubber production by matching tapping intensity to leaf area index

Understanding resource capture can help design appropriate species combinations, planting designs and management. Leaf area index (LAI) and its longevity are the most important factors defining dry matter production and thus growth and productivity. The ecophysiological modifications and yield of rubber (Hevea spp.) in an agroforestry system (AFS) with beans (Phaseolus vulgaris L.) were studied. The experiment was established in Southeast-Brazil, with three rubber cultivars: IAN 3087, RRIM 600 and RRIM 527. The AFS comprised double rows of rubber trees along with beans sown in autumn and winter seasons in 1999. There was about 50% higher rubber yield per tree in the AFS than the rubber monoculture. Trees within the AFS responded to higher solar radiation availability with higher LAI and total foliage area, allowing its greater interception. All three cultivars had higher LAI in the AFS than monoculture, reaching maximum values in the AFS between April and May of 3.17 for RRIM 527; 2.83 for RRIM 600 and 2.28 for IAN 3087. The maximum LAI values for monocrop rubber trees were: 2.65, 2.62 and 1.99, respectively, for each cultivar. Rubber production and LAI were positively correlated in both the AFS and monoculture but leaf fall of rubber trees in the AFS was delayed and total phytomass was larger. It is suggested that trees in the AFS were under exploited and could yield more without compromising their life cycle if the tapping system was intensified. This shows how knowledge of LAI can be used to manage tapping intensity in the field, leading to higher rubber yield.

Nutrition in bean plants and anthracnose intensity in function of silicon and copper application.

Nutrition in bean plants and anthracnose intensity in function of silicon and copper application. The objective of this work was to evaluate the effect of calcium silicate and copper sulfate on anthracnose intensity and nutrition of bean plants. The experiment was conducted using an experimental design in randomized blocks following a 4 x 4 factorial arrangement , (four levels of calcium silicate and four levels of copper sulfate) and two additional treatments (plants without inoculation and plants sprinkled with Benomyl). Four evaluations of the incidence and severity of anthracnose were done, in addition to measuring, total leaf area. At the end of the evaluations, incidence: and data were integrated over time, obtaining the area under disease progress curve (AUDPC). Contents of N, P, K, Ca, Mg, B, Cu, Fe, Mn, Zn, Si and lignin were determined in the aerial Part. A linear decrease of the intensity AUDPC was observed with the increase of the doses of calcium silicate. The severity AUDPC was influenced by the doses of copper, obtaining a reduction of 35% on the higher dosage. The supply of silicon and copper altered the content of the K, mg, S, Zn, Ca and Si in the aerial part of the bean plants.

Effect of Toasting Intensity at Cooperage on Phenolic Compounds in Acacia (Robinia pseudoacacia) Heartwood

The phenolic composition of heartwood from Robinia pseudoacacia, commonly known as false acacia, before and after toasting in cooperage was studied by HPLC-DAD and HPLC-DAD/ESI-MS/MS. A total of 41 flavonoid and nonflavonoid compounds were identified, some tentatively, and quantified. Seasoned acacia wood showed high concentrations of flavonoid and low levels of nonflavonoid compounds, the main compounds being the dihydroflavonols dihydrorobinetin, fustin, tetrahydroxy, and trihydroxymethoxy dihydroflavonol, the flavonol robinetin, the flavanones robtin and butin, and a leucorobinetinidin, none of which are found in oak wood. The low molecular weight (LMW) phenolic compounds present also differed from those found in oak, since compounds with a beta-resorcylic structure, gallic related compounds, protocatechuic aldehyde, and some hydroxycinnamic compounds are included, but only a little gallic and ellagic acid. Toasting changed the chromatographic profiles of extracts spectacularly. Thus, the toasted acacia wood contributed flavonoids and condensed tannins (prorobinetin type) in inverse proportion to toasting intensity, while LMW phenolic compounds were directly proportional to toasting intensity, except for gallic and ellagic acid and related compounds. Even though toasting reduced differences between oak and acacia, particular characteristics of this wood must be taken into account when considering its use in cooperage: the presence of flavonoids and compounds with beta-resorcylic structure and the absence of hydrolyzable tannins.

SENSORY APPROACH TO MEASURE FRAGRANCE INTENSITY ON THE SKIN

Sensory analysis is a precise and descriptive measuring technique to quantify human responses to stimuli. Odor, one of these stimuli, is basically the result of the interaction between a chemical stimulus and the olfactory receptor system, which can be described using a number of different dimensions and measures through different sensory tests: threshold, intensity and quality. To measure fragrance performance on the skin, these parameters are very important, but the main attribute to be evaluated is substantivity, thus the importance of the sensory scale chosen to measure perception, discriminate different intensities and determine the substantivity of the fragrance. Some studies comparing the labeled magnitude scale (LMS) with other magnitude scales and their derivations showed that the use of the LMS scale to measure fragrance intensity could semantically understand the intensity of the stimulus. Tests using this scale confirmed the applicability and efficiency of the LMS. PRACTICAL APPLICATIONS The objective of this article is to review the techniques used to measure odor and fragrance intensities applied on the skin. The review shows general sensory techniques and their goals, the newest olfactory mechanism and its contribution to sensory evaluation and which attributes should be considered to measure odor. Substantivity/retentivity or longevity can be regarded as the most important attributes if you want to measure fragrance performance on the skin. Past studies showed different scales tested to measure odor, and some of them demonstrated that the labeled magnitude scale is very suitable to measure fragrance on the skin.

Influence of storage temperature on cooling intensity of topical emulsions containing encapsulated menthol

The cooling intensity of topical emulsions added with encapsulated or free menthol was evaluated by a screened and trained panel recruited based on the American Society for Testing and Materials method. A sensory panel composed of 10 trained judges performed the evaluation of samples stored at 22 +/- 2C for 24 h and, after 28 days of storage, at 37.0 +/- 0.5C. The obtained data were analyzed by analysis of variance and Tukey`s test. The results showed an increase of cooling intensity as a function of encapsulated menthol concentration. The opposite was observed in samples added with free menthol, which may have caused sensory fatigue. Storage at 37 +/- 0.5C for 28 days had no impact on the cooling intensity of emulsions containing encapsulated menthol, demonstrating high stability and suggesting its application in cooling skin care products. In contrast, emulsions added with free menthol showed a drastic decrease of cooling intensity at 37 +/- 0.5C..

Effects of light intensity and dilution rate on the semicontinuous cultivation of Arthrospira (Spirulina) platensis. A kinetic Monod-type approach

Semicontinuous cultures were carried out at different dilution rates (D) and light intensities (I) to determine the maximum productivity of Arthrospira platensis cultivated in helicoidal photobioreactor up to the achievement of pseudo-steady-state conditions. At I = 108 mu mol photons m(-2) s(-1), the semicontinuous regime ensured the highest values of maximum cell concentration (X(m) = 5772 +/- 113 mg L(-1)) and productivity (P(XS) = 1319 +/- 25 mg L(-1) d(-1)) at the lowest (D = 0.1 day(-1)) and the highest (D = 0.3 day(-1)) dilution rates, respectively. A kinetic model derived from that of Monod was proposed to determine the relationship between the product of light intensity to dilution rate (ID) and the cell productivity, which were shown to exert a combined influence on this parameter. This result put into evidence that pseudosteady-state conditions could be modified according to circumstances, conveniently varying one or other of the two independent variables. (C) 2010 Elsevier Ltd. All rights reserved.

Influence of ammonium chloride feeding time and light intensity on the cultivation of Spirulina (Arthrospira) platensis

This study dealt with the influence of both the feeding time and light intensity on the fed-batch culture of the cyanobacterium Spirulina (Arthrospira) platensis using ammonium chloride as a nitrogen source. For this purpose, a 2 2 plus star central composite experimental design combined with response surface methodology was employed, and the maximum cell concentration (X-m), the cell productivity (P-X), and the yield of biomass on nitrogen (Y-X/N) were selected as the response variables. The optimum values of X-m (1,833 mgL(-1)) and Y-X/N (5.9 gg(-1)) estimated by the model at light intensity of 13 klux and feeding time of 17.2 days were very close to those obtained experimentally under these conditions (X-m = 1,771 +/- 41 mg L-1; Y-X/N = 5.7 +/- 0.17 gg(-1)). The cell productivity was a decreasing function of the ammonium chloride feeding time and a quadratic function of the light intensity. The protein and lipid contents of dry biomass collected at the end of cultivations were shown to decrease with increasing light intensity.

Effects of carbon dioxide feeding rate and light intensity on the fed-batch pulse-feeding cultivation of Spirulina platensis in helical photobioreactor

The behavior of S. platensis was investigated in this study through fed-batch pulse-feeding cultures performed at different carbon dioxide feeding rates (F = 0.44-1.03 g L-1 d(-1)) and photosynthetic photon flux density (PPFD = 80-250 mu mol photons m(-2) s(-1)) in a bench-scale helical photobioreactor. To achieve this purpose, an inorganic medium lacking the carbon source was enriched by gaseous carbon dioxide from a cylinder. The maximum cell concentration achieved was 12.8 g L-1 at PPFD = 166 mu mol photons m(-2) s(-1) and F= 0.44 g L-1 d(-1) of CO2. At PPFD = 80 and 125 mu mol photons m(-2) s(-1), the carbon utilization efficiency (CUE) reached maximum values of 50 and 69%, respectively, after about 20 days, and then it decreased, thus highlighting a photolimitation effect. At PPFD = 166 mu mol photons m(-2) s(-1), CUE was >= 90% between 20 and 50 days. The photosynthetic efficiency reached its maximum value (9.4%) at PPFD = 125 mu mol photons m(-2) s(-1). The photoinhibition threshold appeared to strongly depend on the feeding rate: at high PPFD, an increase in the amount of fed CO2 delayed the inhibitory effect on biomass growth, whereas at low PPFD, excess CO2 addition caused the microalga to stop growing. (c) 2007 Elsevier B.V. All rights reserved.