In general, a role of skeletal stem cells in cancer can be seen a

In general, a role of skeletal stem cells in cancer can be seen as running in parallel with their dual physiological functions — as progenitors of skeletal tissues and as providers and organizers of a microenvironment. The progenitor function comes into play in understanding the origin of primary bone tumors; the non-progenitor functions come into DAPT play in understanding how skeletal progenitors contribute to the establishment of hematopoietic and non-hematopoietic malignancies (bone metastasis). Skeletal stem cells may represent direct progenitors of sarcomas. In spite of the identification of specific molecular pathways underpinning

specific types of bone tumors, classical and predominant (and to some extent, partially obsolete) paradigms of histogenesis of bone tumors have largely remained indifferent to the notion that skeletal tissues emanate from a common progenitor. As a result, classification and textbooks of pathology still identify

primary bone tumors based on their predominant phenotype and/or clinical behavior. However, recent work has highlighted the significance of skeletal progenitor cells for understanding the biology of bone tumors. Transformation of murine bone marrow stromal cells in culture is a far more common event than currently appreciated Lumacaftor (perhaps accounting for some reports of extraordinary numbers of population doublings, mistaken as “self-renewal” in some reports). Screening of multiple murine “MSC” lines by in vivo transplantation assays (conducted to probe their osteogenic capacity) easily reveals their tumorigenic properties (our unpublished results) (Fig.1). The latter, in turn, are easily conceived of as the effect of the known chromosomal instability characteristic of murine cell cultures, at variance with humans. Spontaneous immortalization in cultures of human BMSCs, regardless of sporadic reports

[47], is admittedly an exceptional event, reflecting uncontrolled growth conditions. More importantly, while forced expression of hTERT in human skeletal stem cells can boost their osteogenic capacity [48] and [49], prolonged culturing mafosfamide of hTERT-immortalized human skeletal progenitors results in multiple genetic hits that may culminate with acquisition of full-blown tumorigenesis as assayed by in vivo transplantation [50]. By suggesting that inordinately high rates of proliferation over prolonged time can lead to transformation of skeletal progenitors, these data provide a direct view of sarcomagenesis as related to skeletal stem cells. More specifically, a pathogenetic link between Ewing’s sarcoma (a highly malignant bone tumor, EWS) and skeletal progenitors has been suggested recently.

, 1995 and Kapoor et al , 2012) For example, screening of commer

, 1995 and Kapoor et al., 2012). For example, screening of commercial preparations of lipases for obtaining the best conversion of oil to biodiesel is quite common (Nelson et al., 1996, Shah et al., 2004 and Shah and Gupta, 2007b). Similarly,

lipases from different sources are increasingly screened for obtaining the best yield in a promiscuous reaction (Lai et al., 2010 and Li et al., 2008). Invariably, initial rates are compared before the choice for the best biocatalyst is made. This may not necessarily be the best choice as initial rates are just that: rates yet to be affected by multiple factors which start operating as the reaction progresses selleck compound (see later for a discussion on importance of complete progress curve). However, comparing either initial rates (which has “per mg” of the biocatalyst as a part of its units) or even the conversions and yields from two different commercial preparations is actually comparing apples and pears! Foresti and Ferreira (2005a) have discussed this issue in the context of lipase-catalyzed reactions. However, the points raised have wider implications particularly in the context of industrial buy SCH772984 enzymology. • Strictly speaking, the terms “enzyme”, “lipase” and “protein” are not interchangeable. The first one is the total weight of the preparation; “protein” is the total amount of protein in the preparation on a weight basis, this can constitute

a very small percentage of the total weight of the “enzyme”. Lipase, of Vildagliptin course, refers to the amount of pure lipase present in the “enzyme”. This often is an unknown quantity in a commercial enzyme preparation. It is, however, a common practice to use the term lipase for the total amount of protein, that is, the amount of impure lipase. Often, one has to rely upon the context to understand what may be meant. Foresti and Ferreira (2005a,b) have outlined how to avoid some of the above pitfalls by careful considerations while designing the experiments. The efficiency of the enzymes is generally expressed

in terms of initial rates. This is more or less the norm when the workers describe a more efficient biocatalyst design or formulation for catalysis in low-water media (Straathof and Adlercreutz, 2000 and Vulfson et al., 2001). Engineered enzymes by site-directed mutagenesis or directed evolution are also generally evaluated in terms of initial rates. The initial rate, by definition, is the early initial and linear portion of product concentration vs. time graph. In aqueous buffers, this linearity usually persists to at least till 5% conversion has taken place (Purich, 2010). In low-water media, conversions are much slower and one can observe linearity up to 20–30% of the conversion (Solanki and Gupta, 2008 and Solanki and Gupta, 2011). Reactions which display a lag phase or a burst phase pose problems in accurate estimation of the initial rates.

There are indications, however, that this might be significant T

There are indications, however, that this might be significant. Thus, L-phenylalanine benzyl ester, which was found to reduce sickling, appears to partition into the RBC membrane and non-specifically inhibits transport Selleck Sirolimus systems including the Na+/K+ pump, the cation cotransporters (probably the Na+–K+–2Cl− cotransporter, NKCC) and the anion exchanger (AE1)

whilst also increasing passive cation leaks [12]. No information is available on the aromatic aldehydes. The current results provide the first evidence that o-vanillin directly inhibits the RBC KCC, Gardos channel and Psickle. As reported, o-vanillin was found to increase O2 affinity and inhibit sickling, but their effects on these permeability pathways do not depend on this action. Thus, for KCC and Gardos channel, inhibition also occurred when RBCs were treated with either the sulphydryl reacting reagent NEM or the Ca2 + ionophore A23187, manipulations which bypass any anti-sickling action of o-vanillin. The Na+/K+ pump was also inhibited by o-vanillin. Although this raises the possibility that it acts non-specifically, as suggested for the phenylalanine benzyl esters [12], perhaps by partitioning

into the membrane and destabilising the transporters, the much reduced effect of its isoform, para-vanillin (or usually simply vanillin) argues against this. 5HMF, currently in clinical trials in SCD patients, was different in effect, at least in the transport assays carried out in this work. Nevertheless, selleck screening library present findings indicate that it is possible to design aromatic aldehydes which combine a direct inhibitory effect on HbS polymerisation together with favourable effects on reduction

of RBC permeability to thereby increase RBC hydration. These dual effects may potentiate their ability to ameliorate the complications of SCD. There are no conflicts of interest to declare. AH carried out most experiments with assistance from UMC and OTG. Study was designed by JSG, DCR and ST. Analysis was carried out by AH, UMC and OTG. Manuscript was prepared by JSG, AH and DCR. We thank Action Medical Research and the Medical Research Council for financial support. UMC is supported by a BBSRC studentship. OTG is supported through the generosity of a Yousef Jameel Scholarship and the Cambridge Commonwealth Trust. “
“The clinical manifestations of sickle cell anemia Amisulpride (SCA) include marked phenotypic heterogeneity, involving genetic and environmental factors as well. Fetal hemoglobin (HbF) levels and concomitant α-thalassemia are the two best characterized modifiers of severity in SCA and β-thalassemia. α-Thalassemia modulates SCA by reducing the intracellular concentration of sickle cell hemoglobin (HbS), which decreases HbS polymer-induced cellular damage and in turn ameliorates hemolysis. High HbF levels may reduce SCA severity due to its ability to inhibit HbS polymerization and also reduce the mean corpuscular HbS concentration (reviewed in [1] and [2]).

Certain imaging modalities such as magnetic resonance imaging, tr

Certain imaging modalities such as magnetic resonance imaging, transcranial ultrasound, and single-photon emission computed tomography can be useful in making diagnostic decisions in some cases of PD. Devaki Shilpa Surasi, Patrick J. Peller, Zsolt Szabo, Gustavo Mercier, and Rathan M. Subramaniam The clinical diagnosis of Parkinson disease (PD) is difficult, as several other neurodegenerative and basal ganglia disorders have similar clinical presentations. Dopamine transporter single-photon emission computed tomography has been proposed as possible diagnostic tool to help

differentiate idiopathic PD from essential tremor and other disorders that present with parkinsonian symptoms. In addition, it is valuable in the diagnosis of dementia with Lewy bodies, differentiating Selumetinib ic50 it from other causes of dementia such as Alzheimer disease. Shichun Peng, Doris J. Doudet, Vijay Dhawan, and Yilong Ma This article discusses the current use of PET imaging in the evaluation of dopamine function in Parkinson disease (PD). The article reviews the major radioligands targeting dopaminergic systems in patients with parkinsonian disorders. The primary objective is to show the novel

clinical applications of molecular imaging in the diagnosis and assessment of motor and nonmotor symptoms in PD.

Index  487 “
“Li Q, Zhou XD, Kolosov VP, Perelman JM. Sunitinib molecular weight Nicotine suppresses inflammatory factors in HBE16 airway epithelial cells after exposure to cigarette smoke extract and lipopolysaccharide. Transl Res 2010;156:326-34. In our December 2010 publication in Translational Research, we incorrectly stated that cigarette JAK inhibitor smoke extract (CSE) was “supplied by Professor C-H Cho (Department of Pharmacology, University of Hong Kong).” Although Dr Cho prepared the CSE samples, he did not directly provide us with them. Instead, they were obtained by Dr Zhou who had access to the samples while a research fellow at Hong Kong University from 2005 to 2006. “
“Chronic obstructive pulmonary disease (COPD) is a complex systemic disease, that until recently, was underrecognized, underappreciated, and poorly understood. Bonet first described COPD as early as 1679 when he discussed “voluminous lungs,” and yet it wasn’t until the 1960s that physicians began to create formalized definitions of the clinical syndrome they were encountering.1 These initial definitions focused on either clinical characteristics (such as cough and dyspnea) or anatomic features (such as enlargement of alveolar spaces) and, in some sense, neglected expanded features that could be useful in identifying and understanding the disease.

17 and 18 Consistent with the expected lack of effect on CNS immu

17 and 18 Consistent with the expected lack of effect on CNS immune surveillance owing to the gut-selective blockade of lymphocyte trafficking with vedolizumab,20 and 22 no PML cases have been identified in the vedolizumab development program to date. As of June 27, 2013, there were 3129 patients with CD or UC who had received vedolizumab in 11 clinical studies (including GEMINI 1, 2, 3, and the GEMINI long-term extension study) for a median of 313 days (mean, 481 days; range, 1–1977 days). Accounting for a pharmacologic effect duration of approximately 16 weeks after the last vedolizumab dose, 995 of these patients had been exposed to vedolizumab NADPH-oxidase inhibitor for at least 24 months. If the incidence of PML in patients

receiving vedolizumab was similar to that in patients with multiple sclerosis receiving natalizumab (ie, >1 case in 500 patients) before the application of known risk-stratification factors (ie, therapy duration, previous immunosuppressive use, and JC virus seropositivity),17 it is estimated that 6 to 7 cases would have been seen among vedolizumab-exposed patients. Although no PML cases have been reported in

the integrated vedolizumab safety database, additional longer-term observational data are needed to exclude any potential of developing PML as a result of vedolizumab exposure. In conclusion, vedolizumab was not statistically superior to placebo in achieving clinical remission at week 6 among patients with moderately to severely active CD and previous TNF antagonist failure. Several prespecified outcomes Quizartinib datasheet suggest that vedolizumab may lead to clinical remission in TNF antagonist–naive patients with CD and at 10 weeks in TNF antagonist–failure patients.

These clinically relevant response kinetics have potential implications for bridging induction therapy to vedolizumab maintenance therapy, which has established efficacy, in patients Urease with this lifelong condition. The safety profile of vedolizumab was generally similar to that of placebo in this short-term study and was consistent with that of longer-term vedolizumab use in previous studies. The authors thank all of the investigators and patients who participated in this study; Timothy Leach, MD, for assisting with study oversight; Quintiles, Inc, for medical monitoring; and Whitney Kent for her substantial contribution as the clinical research manager in the operational conduct of the study. Medical writing assistance was provided by Stefanie Dorlas, BMath, of MedLogix Communications, LLC, and was funded by Takeda Pharmaceuticals International, Inc. “
“Event Date and Venue Details from 2013 *65th INTERNATIONAL SYMPOSIUM ON CROP PROTECTION 21 May Ghent, BELGIUM Contact: E-mail: [email protected] Web: *3rd INTERNATIONAL ENTOMOPHAGOUS INSECTS CONFERENCE 02-06 June Orford, QUE, CANADA Contact see: *ANNUAL MEETING CANADIAN PHYTOPATHOLOGICAL SOCIETY 16–19 June Edmonton, ALB, CANADA Info: K.

05) ( Fig  3E) CD31, a vascular cell-specific cell–cell adhesion

05) ( Fig. 3E). CD31, a vascular cell-specific cell–cell adhesion molecule, has been identified to play an important part in the process of angiogenesis. We stained CD31 to investigate

the angiogenesis ability in different transplant sites (Fig. 3C). The quantities of CD31+ blood vessels in various syngeneic grafts were significantly different (P = 0.0002): intra-omental syngeneic grafts had more CD31+ blood vessels than subcutaneous syngeneic grafts (P < 0.05), which had more than orthotopic syngeneic grafts (P < 0.05). The quantities of CD31+ blood vessels in various allografts were also significantly different (P = 0.0093): the quantity of CD31+ blood vessels in EX 527 cost orthotopic allografts was more than heterotopic allografts (P < 0.05), while the quantities were not significantly different between two heterotopic allografts (P > 0.05). Compared with the corresponding syngeneic grafts,

all of the allografts had revascularization at lower level (P < 0.05) ( Fig. 4A). Myofibroblasts with capacity of collagen synthesis are involved in Sirolimus in vivo tissue remodeling. We used α-SMA as a marker for myofibroblasts to determine the fibrosis degrees in transplanted trachea (Fig. 3D). In syngeneic grafts, myofibroproliferation was nearly undetectable during the observation time, whereas allografts had more proliferation of myofibroblasts in lamina propria of transplanted trachea (P < 0.05). The percentages of α-SMA positive area were not significantly different in syngeneic

grafts (P = 0.5278). The percentages were significantly Tangeritin different in allografts (P = 0.0030): The percentages of α-SMA+ area in two different heterotopic allografts were similar (P > 0.05), but significantly higher than orthotopic allografts (P < 0.05) ( Fig. 4B). The optimal tool to study OB pathogenesis, no doubt, is human lung transplantation. However, drawbacks such as sparse OB samples, and difficulties of sampling at various times, in addition to complications after sampling like infections, hamper human lung transplantation to act as a “model”. There is therefore a critical need for some animal models that could elucidate the pathogenesis of OB. Of the different tracheal transplantation models employed in this study, each has obvious advantages and drawbacks [15], and previous investigators have not yet come to a consistent conclusion on which of the transplantation models is more qualified as a model for studying OB. Since evidence is mounting that epithelial damage [16] and [17], immune-mediated tissue injury [18], angiogenesis [19] and [20] and fibroproliferative remodeling [21] may be involved in the development of OB, we compared transplantation models in terms of these hotspot issues in this study. In addition, we combined transplantation models to decrease the consumption of the animals as well as improve individual error and the experimental efficiency.

3) Traditionally, a limited set of technologies has been used to

3). Traditionally, a limited set of technologies has been used to characterize micro- and nano-vesicles with more techniques being available to the micro sized particles [32].

These include: flow cytometry, dynamic light scattering (DLS), electron microscopy [33] and [34] or enzyme linked immune-sorbent assays (ELISA) [35], [36] and [37]. Most widespread is flow cytometry; commercial flow cytometry typically has a lower practical size limit (for polystyrene beads) of around 300 nm at which point the signal is indistinguishable from the baseline noise level. Whilst this detection limit can be extended with the use of fluorescent labels, at lower sizes the ability to accurately size such particles is quite limited. Dynamic light scattering has also been

used in this application, but being an ensemble measurement, the results comprise either a simple z-average (intensity weighted) particle size and polydispersity, or a very limited-resolution particle Epacadostat size distribution profile [38] and [39]. Electron microscopy is a useful research tool for studying micro- and nanovesicles but at the expense of capital running costs, extensive sample preparation [40]. Most frequently, EVS are counted in biological samples by flow cytometry [21], [41] and [42]. Several authors have pointed out that despite flow cytometry is the technique of choice for evaluation of EVS, it is limited by lack of adequate standardization [43]. Preanalytical as well as analytical issues have been evaluated in detail by Yuana et al. [44]. Preanalytical parameters Y-27632 mouse were also studied in our laboratory, when measuring EVS in stored blood products: in addition to the flow cytometry parameters chosen for the numbering of EVS, we observed that many preanalytical variables have to be taken into account (diluents used,

temperature, vortexing duration, etc.) [45] and [46]. In addition, Mullier et al. recently evaluated many aspects of the prenalytical conditions as well as pending issue that has to be considered when measuring EVS in blood samples Liothyronine Sodium [47]. Because REVS express transmembrane proteins such as band 3, glycophorins, or blood group antigens, it is quite convenient to use specific antibodies raised against glycophorin A, CD47 or other blood group specific antigens for flow cytometric purposes. The expression of negatively charged phospholipids (surface phosphatidylserine) at the external part of the vesicles membrane can be explored by using annexin V as a ligand. However, standardization is mandatory in order to be able to compare data from different sets of experiments, to compare normal individuals with patients and to compare data from different laboratories. Recently, Xiong et al. presented a standardized approach based on quantitative flow cytometric technique [48]. The enumeration of REVS is made possible by using a fixed number of different-sized calibration beads spiked into each sample.

’ [31] Annex III of the OSPAR Convention was also amended to ena

’ [31]. Annex III of the OSPAR Convention was also amended to enable, on the same conditions set out in Annex II, the dumping of CO2 streams from offshore installations. The EU CCS Directive establishes a detailed legal framework for the environmentally safe storage of CO2 both onshore and offshore. The UK has implemented (‘transposed’) the Directive׳s

provisions by modifying its pre-existing petroleum legislation and associated regulatory policies [32]. Existing UK legal and policy frameworks that impact on offshore CO2 storage and planning for such activities fall into four broad clusters, which are discussed below: This legislation was developed in order to consolidate regulation and AZD2281 order planning of marine activities in UK waters, and implement in a marine context the UK Government׳s commitment to sustainable development [33],

[34], [35], [36] and [37]. The Act׳s core provisions relate to: establishment of the Marine Management Organisation (MMO) (Part 1); designation of certain maritime zones (Part 2); marine planning and licensing (Parts 3 and 4); nature conservation including the designation of marine conservation zones (Part 5); inshore and offshore fisheries management (Parts 6 and 7); law enforcement (Part 8); and recreational coastal access (Part 9). The foundation of the Act׳s marine planning and licensing framework is a ‘Marine Policy Statement’, in which the UK Government and other participating government bodies publish general policies ‘for contributing to the achievement of sustainable development’ in UK waters [38]. The current (and first) Marine Policy Statement buy PD0332991 was published in March 2011 and Staurosporine manufacturer was prepared jointly by the UK Government, Northern Ireland

Executive, Scottish Government and Welsh Assembly Government [39]. The statement contains several paragraphs that highlight the importance of offshore CO2 storage, and planning for such activities, as means of implementing the UK׳s legal and policy commitments concerning climate change mitigation [40]. The MCAA subdivides UK waters into eight ‘marine planning regions’ which correspond to the inshore and offshore regions of England, Northern Ireland, Scotland and Wales [41]. The Act does not establish a planning framework for the inshore regions of Northern Ireland and Scotland, reflecting a devolution of legislative responsibility to those constituent countries [42]. For each of the remaining six planning regions (or parts thereof), the Act provides for the preparation of a ‘Marine Plan’ by designated government bodies [43]. The list of designated bodies includes the MMO, which operates autonomously from the UK Government, but is required to comply with directions issued under with MCAA section 37 by the Secretary of State (i.e. cabinet minister) in charge of the UK Department of Environment, Food and Rural Affairs (DEFRA).

Fortunately, the identification of molecules using mass spectrome

Fortunately, the identification of molecules using mass spectrometry analysis is helping to characterize these neglected molecules and change the current scenario [14] and [15]. Through natural selection, scorpion venoms molecules were conserved to act upon certain physiological mechanisms which are shared by a great variety of organisms, including

human beings. Therefore, it is probable that compounds like scorpion venom peptides can be prototypes for the development of new drugs. For example, the chlorotoxin (CTX) from the scorpion Leiurus quinquestriatus was first described as a chloride toxin [8], but nowadays it has been shown to be effective against the human glioma brain tumor via inhibition of the MMP-2, an important metallopeptidase over-expressed by tumor cells [22]. This fact Ibrutinib molecular weight suggests that novelties are still to be discovered, including new functions for already known molecules. Thimet oligopeptidase (EP24.15) belongs to the M3 family metallopeptidase Dinaciclib research buy [13] and was first described as a neuropeptide-degrading enzyme present in the soluble fraction of brain homogenates [12]. The EP24.15 does not have a clear primary

specificity to cleave substrates, with the ability to accommodate different amino acid residues at subsites S4 to S3′ [11]. In fact, EP24.15 shows substrate size restriction to peptides containing from 5 to 17 amino acids because of its catalytic center, located in a deep channel [17]. These features of EP24.15 were decisive in successfully describing two new peptides: the human hemopressin [19] and a potent inhibitor of ACE in the venom of Bothrops

jararacussu [20]. Considering the property of EP24.15 to select small molecules and its presence in the nervous system, where the TsV mainly acts, the objective of this study was to find in TsV new bioactive peptides selected by interaction with EP24.15 activity in vitro. The lyophilized TsV, provided by Butantan Institute, São Paulo, Brazil, was suspended in sodium acetate pH 4.0 and immediately fractionated at 4 °C using a 10 kDa molecular weight cut off membrane (Millipore), ifenprodil in order to prevent proteolytic cleavage of peptides by the crude venom. The filtrated solution (Peptide Pool) was subjected to reverse phase HPLC (Prominence, Shimadzu), using a Shim-pack VP-ODS C-18 column (4.6 × 150 mm); 0.1% TFA in water (solvent A), and acetonitrile plus solvent A (9:1) as solvent B. The chromatography was performed at a flow rate of 1 mL/min and detected by ultraviolet absorption (214 nm). The peaks were collected manually, dried and subjected to enzymatic assays. The recombinant EP24.15 was obtained as described [18]. The peptidase assay was conducted in a 50 mM phosphate and 20 mM NaCl 7.

1 M HEPES/NaOH (pH 8 5) and 25 μL of NaCl solution (6 mM–1 2 M) i

1 M HEPES/NaOH (pH 8.5) and 25 μL of NaCl solution (6 mM–1.2 M) in a micro centrifuge tube. The reactions were initiated by the addition of 25 μL of the midgut homogenate to the tubes, and the mixtures were

then incubated at 30 °C for 2 h. The reducing carbohydrates released from the substrate by the action of the amylase were quantified using the dinitrosalicylic acid method, as described in Section 2.2.1. The blanks were prepared with the same NaCl concentrations and with water instead of samples. The assays in the absence of Cl− were performed separately using a similar protocol. The dissociation constant of the Cl− ion from the amylase was calculated using GRAFIT (Erithacus Software, version 7.0), assuming the enzyme was saturated with the substrate. To investigate the influence of calcium ions, 10 total midguts were dissected in 0.9% (w/v) NaCl and transferred to 250 μL of 600 mM NaCl. The samples were homogenized using an abrasive micro-homogenizer Selleck Bortezomib made of

glass and then centrifuged at 4 °C for 10 min at 14,000×g. The supernatant containing the equivalent of 1 midgut (25 μL) was used in the assays. The assays where performed mixing 100 μL of a 1.5% (w/v) aqueous starch solution, 150 μL of 0.1 M HEPES/NaOH buy Regorafenib (pH 8.5) and 25 μL of different CaCl2 solutions (concentrations varying from zero to 96 mM) in a micro centrifuge tube. The reaction was started by the addition of 25 μL of the sample, and the tubes were incubated at 30 °C for 1 h. The reducing carbohydrates released from the Methocarbamol substrate were quantified using the dinitrosalicylic acid method, as described in Section 2.2.1. The blanks were prepared with the same CaCl2 concentrations and with water in the place of sample. The midgut sample containing amylase was obtained by homogenizing 5 total midguts in 50 μL of 200 mM

NaCl. After centrifugation at 14,000×g at 4 °C for 10 min, the supernatant was used for the starch hydrolysis assay. The starch hydrolysis was assayed by mixing 100 μL of a 4.5% (w/v) aqueous starch solution with 150 μL of 0.1 M HEPES/NaOH buffer (pH 8.5) and 50 μL of a sample containing the equivalent of 5 midguts in a micro centrifuge tube. The mixture was incubated at 30 °C for 6 h. Throughout the incubation time, 20 μL aliquots were collected at 0, 1.5, 3 and 6 h and transferred to another tube in which the action of the amylase on starch was inactivated by immersion in boiling water for 2 min. All three samples were centrifuged (14,000×g, 10 min), and 15 μL from each aliquot was applied to a silica gel plate (Fluka 99903). The chromatography was performed using a mixture of butanol, ethanol and water (5:3:2, v/v/v). The spots corresponding to the products of starch hydrolysis were developed via aspersion of an ethanol/sulfuric acid mixture (9:1) and heating at 100 °C in an oven. The processivity of the α-amylase-starch complex was evaluated according to the method of Robyt and French (1967) and Bragatto et al. (2010) with modifications.