The environmental conditions, pigment characteristics, growth act

The environmental conditions, pigment characteristics, growth activity etc., relating to the bloom are described in detail elsewhere ( Furuya et al.

2006). The primary Quizartinib objective of this work is to describe the phytoplanktonspecific absorption characteristics of the bay during the bloom. Secondly, an attempt is made to identify the pigments responsible for the major absorption peaks by resolving the overlapping features in the absorption spectra through derivative analysis. Samples were collected during fieldwork carried out in Manila Bay from 19 to 23 March 2004. The stations were distributed along two transects: an east-west (EW) transect between Manila and Limay (stn. MB7–13) and a north-south (NS) transect from the mouth of the bay to Pampanga (stn. MB1–5, MB10 & 11) (Figure 1). Physical parameters like temperature, salinity and conductivity were obtained using a portable CTD profiler. Samples for phytoplankton composition based on HPLC and phytoplankton this website spectral absorption were collected from different depths down to 23 m using a

Nansen sampler; surface (~ 5 cm) sampling was done using a bucket. Seawater samples (0.5–1 litres) were filtered onto 25 mm GF/F glass fibre filters under low vacuum pressure (< 25 hPa). The absorption spectra of total particulate matter was recorded in the wavelength range 350–750 nm at a resolution of 1 nm with a double-beam spectrophotometer (Shimadsu

MPS-2400) following the guidelines of Mitchell (1990). For each of the measured spectra, the optical density obtained at 750 nm was subtracted from all other wavelengths. The optical density of the total suspended matter was corrected for the path length amplification (β effect) according to Cleveland & Weidemann (1993). The optical density of detritus particles was measured following the pigment extraction method selleck chemicals llc of Kishino et al. (1985). The chlorophyll-specific absorption coefficients of phytoplankton (a*ph(λ)) were obtained by dividing the absorption coefficient of phytoplankton (aph(λ)) by the total Chl a (TChl a) concentration. TChl a and TChl b includes both mono and divinyl forms. Biomarker pigments were separated and quantified using reverse-phase gradient elution HPLC following Zapata et al. (2000). Seawater was filtered under a gentle vacuum (< 100 mm Hg) onto 25 mm glass fibre filters (Whatman GF/F) and stored immediately in liquid nitrogen. Pigments were extracted using methanol (95%), and the extract was mixed with 1 M ammonium acetate as the ion pairing reagent. It was then filtered through 0.2 μm PTFE filter (Whatman) and mixed with milli-Q water (5:1 v:v); thereafter 500 μl was injected into the HPLC system (Shimadzu) equipped with a Symmetry C8 column (Waters).

Huang-Hua-Zhan (HHZ), a high yielding indica variety from South C

Huang-Hua-Zhan (HHZ), a high yielding indica variety from South China was used as the recurrent parent (RP) to cross with three donors, IR64 (indica from IRRI), AT354 (indica from Sri Lanka) and C418 (japonica from Northeast China). The F1s were backcrossed to HHZ to produced 3 BC1F1 populations, from each of which 20–25 random BC1F1 plants were further Ipilimumab backcrossed to HHZ to produce 20–25 BC2F1 lines.

The selfed seeds from all BC2F1 plants from a single cross were bulk harvested, forming a single BC2F2 population. The BC2F2 populations were subjected to three different selection schemes for improving the target traits (ST, HY and DT), as described in Fig. 1. Three different selection schemes were adopted in this study. In the first selection scheme, each of the bulk BC2F2 populations was screened for DT under natural drought stress conditions (the soil type is sandy yellow clay) at the CAAS experiment station in Hainan during the 2009–2010 dry season (DS). Seeds of the bulk BC2F2 populations Selleck I BET 762 were sown in the nursery on November 20, 2009, and 400 25-day old seedlings

of each BC population were transplanted into a 40-row plot with one row of HHZ inserted every 20 rows as checks. The spacing was 20 cm × 17 cm. Drought stress was started by draining the field at the peak tillering stage 30 days after transplanting. One irrigation Ribonuclease T1 was applied to prevent death when drought stress was severe at 65 days after the stress started. At the maturity, 19, 29 and 33 plants that had obviously higher yield than HHZ were visually selected from the HHZ/IR64, HHZ/AT354 and HHZ/C418 populations. The 81 DT selected HHZ BC2F3 introgression lines (ILs) and HHZ were progeny tested under both irrigation and drought stress conditions at the CAAS experimental station in Beijing in the

2010 summer. Seeds of each IL were sown into a seeding tray and 45 30-day old seedlings of each IL were transplanted into a 3-row plot with a spacing of 20 cm × 17 cm and two replications for each IL and HHZ under each water treatment. In the irrigated control, a 5 cm layer of standing water was maintained in the field until 10 days before harvest. The crop management was the standard one with two applications of fertilizers at a total rate of 120 N kg ha− 1. Under the drought treatment, all ILs were evaluated in the greenhouse at the CAAS experimental station with the same experiment design. A terminal drought was initiated by withholding irrigation 30 days after transplanting until maturity, drought conditions that were very severe killing HHZ and some ILs. However, 43 ILs survived the stress and were able to produce seeds. These included 12 ILs from the HHZ/IR64 population, 23 ILs from the HHZ/AT354 population, and 8 ILs from the HHZ/C418 population (Fig. 1).

New strategies including coupling or integration of


New strategies including coupling or integration of

complementary processes are necessary to establish economical and efficient industrial scale processes not only for fractionation, but also for simultaneous and continuous production of peptides with different bioactive properties. For example, Wu et al. [13] reported that an enzymatic ultrafiltration (UF) membrane reactor in conjunction with chromatography could be used to achieve continuous hydrolysis and isolation of multi-functional peptides more effectively than the traditional mode using batch reactors. The selection of membranes with appropriate molecular weight cut-off followed by either size-exclusion chromatography or cation exchange chromatography enabled simultaneous production and isolation GSK1120212 in vivo of peptides with ACE-inhibitory, calcium-binding and antimicrobial properties [13]. In recent years, a process coined ‘EDUF’ for ‘electrodialysis with UF membranes’ has been explored to separate molecules by electric charge and molecular mass 14 and 15•. In EDUF, the driving force through the membranes is via an electric field (anode/cathode) rather than by pressure as is the case

with conventional UF, thus mitigating the limitations of both chromatography selleck (high cost) and pressure-driven membrane (fouling) processes. EDUF can be used to achieve simultaneous production and fractionation in a single step, and the higher resolution achieved by stacking differently sized UF membranes can result in purified peptide fractions with higher functionality and bioactivity

[15•]. Bioinformatics, also known as in silico prediction and analysis, refers to computational methods applied to manage, curate and interpret information on biological systems, in this case, the bioactive peptides derived from food. Based on knowledge about structure and activity of peptides reported GPX6 in the literature and deposited in pertinent databases, computational approaches may be applied to elucidate structure–function relationships, predict peptide sequences likely to exhibit specific activities, locate peptides encrypted in particular protein sources, envisage release of those fragments by specific enzymatic cleavage, and propose the putative mechanism of action through molecular docking of binding sites [16]. Although there is a growing bank of databases pertinent to bioactive peptides and the proteolytic enzymes that may be used to release them from food proteins [17], the majority describe bioactive peptides found endogenously, that is, of physiological relevance, rather than being derived from food.

No entanto, não existem, até à data, dados suficientes que fundam

No entanto, não existem, até à data, dados suficientes que fundamentem a utilização destes parâmetros como indicadores de inflamação eosinofílica da doença4. A demora que normalmente existe entre o início dos sintomas e o diagnóstico é em média 4,3 anos (1-13 anos). A EEo tem um caráter crónico e recidivante, sendo a atividade da doença muito variável. Tem sido sugerida uma flutuação da atividade da doença see more dependente da exposição a aeroalergénios, nomeadamente pólenes15. Podem surgir complicações, nomeadamente alterações

estruturais como fibrose ou estenose que podem ser irreversíveis, bem como alterações funcionais. Até à data, não houve associação a neoplasias malignas25. A importância de tratar estes doentes prende-se com 3 vertentes: melhoria da qualidade de vida, diminuição do risco de lesões esofágicas graves que levem ao impacto alimentar e prevenção da lesão Sunitinib mouse do órgão causada pelo remodeling tecidular 26. O tratamento incide na dieta alimentar, tratamento farmacológico e tratamento endoscópico. Existem 3 tipos de dietas de evicção: dieta de evicção dos alimentos reconhecidos como mais alergénicos tais como leite, ovo, peixe, marisco, frutos secos, amendoim, soja e trigo

(eficácia 74%), a dieta orientada pelos resultados da avaliação alergológica (eficácia 76%) e a dieta elementar, baseada numa fórmula de aminoácidos (eficácia 88 a 100%)13, 21 and 27. Nos últimos anos, tem-se demonstrado a eficácia clínica e histológica destas dietas, sobretudo nas crianças28. No entanto, num estudo realizado em adultos, verificou-se que a dieta de evicção dos alimentos reconhecidos como mais alergénicos tinha uma eficácia de 78%22. A dieta elementar, aplicável habitualmente

nas crianças, apesar de ser a mais eficaz, é aquela que é mais difícil de cumprir. Por um lado, pelas restrições alimentares subjacentes e, por outro, pela necessidade de ingestão de grandes volumes de fórmulas Fludarabine price elementares, para que não surjam défices calóricos/nutricionais. A dieta de evicção dos alimentos reconhecidos como mais alergénicos e a dieta orientada pelos resultados da avaliação alergológica são mais práticas29. No entanto, a primeira, dada a grande diversidade de alimentos a evitar, condiciona uma dieta muito restritiva e, eventualmente, desnecessária, podendo também condicionar deficiências nutricionais. Além disso, a eficácia parece ser ligeiramente superior para a dieta orientada pelo estudo alergológico (76 versus 74%), como referido anteriormente. Após a remissão da doençam os alimentos devem ser reintroduzidos na dieta de forma gradual, mantendo aqueles que não levam a recorrência29. A evicção prolongada de alimentos para os quais existe uma sensibilização assintomática, pode levar à ocorrência de reações sistémicas IgE mediadas, aquando da sua reintrodução na dieta.

The timing of encoding-related brain activity observed here is al

The timing of encoding-related brain activity observed here is also consistent with the involvement of a preparatory process. The activity started around 1 sec after cue onset and ended just before word onset, similar to what has been observed previously when the input modalities of the cue and word are kept constant (Otten et al., 2010). The relatively late onset of the effect points to a preparatory process engaged in anticipation of the upcoming event rather than a cue-specific process. Interestingly, we observed

an additional Idelalisib prestimulus effect for auditory words. While the negative frontal effect occurred prior to visual and auditory words, a more posteriorly distributed effect was observed for auditory

words in the easy cue discrimination condition. Activity shortly after the onset of auditory cues was more positive when the following word was later recalled. This effect was maximal over posterior scalp sites, suggesting a contribution of the P300 family of components (Donchin and Coles, 1988). Given the suggested role of the P300 in context updating and working memory, this might not seem surprising. The information about the upcoming input modality delivered by the cues is highly relevant and the better this information is processed, the more effective preparation might be. However, there seems little reason to assume why this would only be relevant for words presented in the auditory modality. We have previously noted that auditory words are special Selleck Ivacaftor in the learning of short word lists (Galli et al., 2012). Anacetrapib The same conclusion is evident from the fact that faster cue discrimination times increased likelihood of recall

for auditory words, whereas recall was less likely for visual words. A special status of auditory information is also apparent from the simple discrimination tasks we gave participants. When visual gratings and auditory tones were presented in isolation, speed of discrimination was identical. This means that discriminations were not inherently easier for one or the other input modality. However, as soon as gratings and tones were presented in the same temporal sequence as used during memorization, discrimination times were slower for auditory decisions even though no words were presented. Although it is not clear how this translates to the positive prestimulus effect seen for auditory words, auditory processing must be especially sensitive to the temporal dynamics of the sequence in which stimuli are embedded. Importantly, the fact that this type of prestimulus activity was again only observed during the easy discrimination task emphasizes the importance of processing resources in the elicitation of prestimulus activity. Brain activity after word onset was also predictive of subsequent memory performance.

All patients treated at our institution and for

whom medi

All patients treated at our institution and for

whom medical records were available were selected for inclusion in this retrospective study. Initial locoregional staging included a clinical evaluation performed by a gynecologic surgeon and radiation oncologist (according to the 1995 FIGO (International Federation of Gynecology and Obstetrics) classification (7)). Abdominopelvic MRI was obtained for 168 patients (74.3%) and CT imaging for 160 patients (70.8%). FDG-PET (fluorine-18-fluorodeoxyglucose positron emission tomography) scan was not systematically Selleckchem Sirolimus performed, and no decision has been taken based only on its results; 148 patients (65.4%), mostly Stages I and II, with a good health status and without suspicious lomboaortic nodes at CT or MRI were selected to receive pelvic lymphadenectomy by coelioscopy first. Only 1 patient had a para-aortic lymphadenectomy. Nodal involvement was determined if histologically proven (65 patients) or suspected on CT (24 patients). All patients with positive lymph nodes, including IB1 stage, received first external beam radiation therapy and are included in the study. Stage IB1 patients treated with preoperative intracavitary PDR BT followed with colpohysterectomy were excluded from the analysis (19 patients). Institutional review board approval

was obtained for this study, and it was conducted in compliance with the Helsinki Declaration. All patients received 45 Gy pelvic external beam radiotherapy (EBRT) before PDR BT with a standard four-field technique (190 patients) EGFR inhibitor or a two anterior/posterior opposing fields technique (36 patients) using high E7080 ic50 megavoltage photons from a linear accelerator (photons × 18 and 25 MeV). EBRT included the para-aortic area when the CT showed enlarged common iliac or para-aortic nodes. When the nodal involvement was histologically proved or suspected on CT, a complementary boost irradiation was delivered after BT to reach a minimum of 60 Gy to the parametria and/or involved pelvic nodes and 55 Gy to the para-aortic

nodes, taking into account the dose contribution of BT. From 1999, based on the results of randomized trials [8], [9], [10], [11] and [12], chemotherapy was given during EBRT for all stages ≥IB2, with intravenous cisplatin 40 mg/m² once a week for 5 weeks in 150 of 226 patients (66.4%). Chemotherapy courses were not delivered during the hospitalization for the BT procedure. After EBRT, the PDR BT boost was delivered during a single hospitalization, using the PDR Selectron (Elekta, Stockholm, Sweden). At the beginning of the BT procedure, a careful clinical examination was carried out under general anesthesia to assess clinical response to EBRT. A Fletcher applicator was used, and no patient underwent interstitial BT. Pulses were delivered hourly during night and day. Before 1999, the BT treatment planning dosimetry was based on orthogonal radiographs, in accordance with International Commission on Radiation Units (ICRU) 38 (13).

Peers may have the potential to influence health outcomes of othe

Peers may have the potential to influence health outcomes of other patients by addressing feelings of isolation, promoting a positive outlook, and encouraging healthy behaviour [16]. A better understanding of what actually takes place in peer support interventions is needed, to tease out how peer support works, in what circumstances and for whom. This paper

synthesizes Histone Methyltransferase inhibitor qualitative research about the experiences and perceived impacts of peer support interventions across multiple chronic diseases, and in so doing, works towards a conceptual model. It also aims to identify both positive and negative aspects of peer support, and examine which experiences and perceived impacts have relevance for mentors and mentees. Given the growing interest in developing evidence based peer support interventions for people with chronic illness [17], it is important to build on what is already known. We aim to contribute to the development and implementation of future interventions. The technique of meta-ethnography was chosen for qualitative data synthesis as it is an interpretive method that preserves the qualitative Selleck DAPT nature of the material being synthesised [18]. Meta-ethnography encourages a clearer understanding of how concepts in different studies are related to each other. This mutual “translation” preserves the structure

of relationships between concepts within any given study, thereby reducing the possibility of de-contextualization [19]. The value of meta-ethnography lies not only in its ability to retain the meaning of primary data, but also in its potential to enable a higher level of analysis and generate new conceptual models. Meta-ethnography requires a literature search strategy, abstract selection, quality

appraisal, and extraction, translation, and synthesis of concepts [19]. These stages were carried out by a team of 17 researchers including two people with arthritis (one of the chronic diseases included in the synthesis). Regular face to face, tele- and video-conference meetings were held over 30 months. A customized web-based platform facilitated data extraction and analysis of the identified articles. Seven comprehensive, on-line literature Fluorouracil searches were conducted across the following disease categories: rheumatic disease, HIV/AIDS, cardiovascular disease (CVD), cancer, asthma, diabetes, and chronic disease. These diseases were identified by team consensus and by a desire to focus on physical diseases. Searched databases included MEDLINE (Ovid SP), EMBASE (Ovid SP), CINAHL (EbscoHOST), PsycINFO (Scholars Portal), ERIC (Scholars Portal), Social Sciences Citation Index (Scholars Portal), Social Work Abstracts (Scholars Portal), Cochrane Database of Systematic Reviews, The Cochrane Library (Wiley Interscience), and DARE (Centre for Reviews and Dissemination). There were no date restrictions. Studies were published in English.

Water, bile, enzymes, and mucous contribute to the change in cons

Water, bile, enzymes, and mucous contribute to the change in consistency. Once the nutrients have been absorbed and the leftover-food residue liquid has passed through the small intestine, it then moves onto the large intestine for expulsion. Lastly bile toxicity has been related to apoptosis and necrosis, not bacteria infection which has been demonstrated in several studies. The absence of I-BET-762 chemical structure evidence of disease transmission to other reef organisms is promising for field testing.

The doses of oxbile required to kill A. planci, concentration 4 g l−1 at 10 ml volume (single injection) are very low compared to doses used when injecting sodium bisulfate, 140 g l−1 concentration at 60 ml volumes (multiple injections) ( Kayal etal., 2011). The amount of oxbile 5-FU purchase injected is only 0.04 mg/sea star which is distributed in A. planci tissues and it will be attacked, englobed and partially degraded by the sea star immune cells and expelled through the water vascular system as part of the regular functions of the immune system. More importantly, many bacteria are capable of transforming and degrading bile in the digestive tract and in the environment ( Hofmann

and Hagey, 2008 and Bodo, 2011). Bacterial bile salt transformation and degradation is of high ecological relevance and also essential for the biotechnological production of steroid drugs (Bodo, 2011). Thus, A. planci remains containing bile salts will be constantly degraded by different mechanisms. Normally, considerable amount of bile salts is released into the environment Exoribonuclease with faeces and urine of

vertebrates. Bile salts cholate, glycocholate, deoxycholate and glycodeoxycholate are also produced and degraded by marine bacteria ( Bode et al., 2003, Maneerat et al., 2005 and Kim et al., 2007). Moreover, aerobic bacteria are able to grow with bile salts as sole source of carbon and energy. For energy conservation, these bacteria oxidase steroid compounds completely to CO2. In the water column, petromyzonol sulfate which is the major bile salt in sea lampreys is subject to microbial degradation ( Hagey et al., 2010). On the GBR, eradication of outbreaks populations of A. planci are predicted to reverse the current trend of declining coral cover ( De’ath et al., 2012). The low doses (concentration and volume) and limited risk of unintended casualties make oxbile and oxgall good candidates for field testing as a novel control method for A. planci. This new approach, coupled with strategic measures to improve water quality, could mitigate the effects of A. planci on coral communities and enable gradual recovery of coral assemblages and reef ecosystems. “
“The authors wish to correct the accidental omission of three of the author names from their poster abstract printed in J Am Med Dir Assoc 2013;14:B17-B18. The Author line should be corrected to: “Author(s): Renee M.

Plasma creatine kinase activity was determined using the CK-UV Ki

Plasma creatine kinase activity was determined using the CK-UV Kit (Bioclin, Brazil). Activity was expressed in units/L, with one unit corresponding to the production of 1 μmol of NADH per min at 30 °C. Negative controls received 50 μL of 1% DMSO-PBS alone. The control group for each sesquiterpene lactone compound received an intramuscular PD98059 injection of 50 μL of a solution containing

just 20 μg of each sesquiterpene lactone derivative compound, dissolved in 1% DMSO in PBS (pH 7.2) ( Souza et al., 2008 and Da Silva et al., 2008a). The measurement of the enzymatic activity using the micellar substrate, 1-hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphoglycerol (HPGP), was performed through the microtiter plate assay. Each sesquiterpene lactone derivative compound was tested in final concentrations of 0.0, 1.0, 2.0, 3.0 and 4.0 μM. Seven wells of a 96-well microtiter plate were used for each assay, resulting in six measurement repetitions of the enzymatic activity for each final concentration of the inhibitor. Thus, for each assay with different concentrations of the inhibitor, 100 μL of solution A in assay buffer (27 μM bovine serum albumin, 50 mM KCl, 1 mM CaCl2, 50 mM Tris– HCl, pH 8.0) was added to seven wells, followed by the addition of a volume of lactone OSI-744 clinical trial derivative compound (the volume used was according to the assay concentration,

0.0–4.0 μM) dissolved in DMSO. For control reactions, the same volume of DMSO was used Methane monooxygenase alone. Solution B had the same composition as that of Solution A, but contained PLA2 (0.5 μg/mL) and was delivered in 100 μL volumes to seven wells, except for the first well. Instead of Solution B, an additional

100 μL-portion of Solution A was added to the first of the seven wells in the assay. Solution B was prepared immediately before each set of assays to avoid loss of enzymatic activity. Quickly, after the addition of Solution B, the assay was initiated by the addition of 0.5–50 μL of Solution C to the seven wells (53 mM HPGP vesicles in assay buffer), with a repeating pipette. The final concentration of HPGP varied from 0.125 to 10 mM. The final volume of the assay was 265 μL and, when necessary, the wells received an extra volume of solution A in order to complete this volume. The fluorescence (excitation = 342 nm, emission = 395 nm) was read with a microtiter plate spectrophotometer (Fluorocount, Packard Instruments). Control reactions without enzyme or inhibitor were run for all assays and the initial velocity was calculated from the initial slope of fluorescence versus time, for each concentration of the substrate used. The significance of differences between groups was evaluated using the Student’s t-test. A P-value <0.05 was considered to be significant ( Da Silva et al., 2008b). The structures of each sesquiterpene lactone derivative compound were submitted to ab initio quantum calculations. In order to select the best conformations, the HyperChem 7.51 software was utilized.

Our results indicate that pro-oxidant concentrations of retinol i

Our results indicate that pro-oxidant concentrations of retinol induce the activation of redox-sensitive pathways which result in the up-regulation of RAGE in cultured Sertoli cells. Pregnant Wistar rats were housed individually in Plexiglas cages. Litters were restricted to eight pups each. Animals were maintained selleck inhibitor on a 12-h light/dark cycle at a

constant temperature of 23 °C, with free access to commercial food and water. Male immature rats (15 days old) were killed by cervical dislocation. All procedures were approved by the Local Ethics Committee Board of UFRGS. All-trans retinol alcohol, Trolox, 2′,7′-dichlorohydrofluorescein diacetate (DCFH-DA), 3-(4,5-dimethyl)-2,5-diphenyl tetrazolium bromide (MTT), Tween-20, and β-mercaptoethanol were from Sigma Chemical Co. (St. Louis, MO, USA). Retinol was dissolved in ethanol. Concentrated stocks were prepared immediately before experiments by diluting retinol into ethanol and determining final stock concentration by UV absorption; solution was kept protected from light and temperature during all procedures. Appropriate solvent controls were performed for each condition. Treatments were initiated by adding concentrated solutions to reach final concentrations

in the well. The final ethanol concentration did not exceed 0.2% Gefitinib purchase in any experiment. Tissue culture reagents were from Gibco (Invitrogen Corporation, Carlsbad, CA, USA) and were of tissue Inositol monophosphatase 1 culture grade. Rabbit polyclonal anti-RAGE was from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse monoclonal anti-β-actin was from Sigma. Rabbit polyclonal anti-p38 (phosphorylated form) and anti-Akt (phosphorylated form) were from Santa Cruz. Sodium dodecyl sulphate (SDS)–polyacrylamide gel electrophoresis (PAGE) reagents were from Bio-Rad Laboratories (Hercules, CA, USA), nitrocellulose membrane (Hybond ECL), enhanced chemiluminescence kit (ECL plus), and anti-rabbit immunoglobulin (horseradish peroxidase-linked whole antibody from donkey) were from Amersham Pharmacia Biotech (Amersham,

UK). UO126 was from Promega Corporation (Madison, WI, USA), GÖ6983 and SB203580 were from Merck Biosciences (Darmstadt, Germany) and H89 was from Biomol Research Laboratories (Plymouth Meeting, PA, USA). Other kinase inhibitors were a kind gift from Professor Peter Dunkley (University of Newcastle, NSW, Australia). Sertoli cells were isolated as previously described (Pasquali et al., 2008). Briefly, testes of 15-day-old rats were removed, decapsulated and digested enzymatically with trypsin and deoxyribonuclease for 30 min at 37 °C, and centrifuged at 750 × g for 5 min. The pellet was mixed soybean trypsin inhibitor, then centrifuged and incubated with collagenase and hyaluronidase for 30 min at 37 °C. After incubation, this fraction was centrifuged (10 min at 40 × g). The pellet was taken to isolate Sertoli cells and supernatant was discarded.