The Rayleigh resolution of a zone plate TXM system is determined

The Rayleigh resolution of a zone plate TXM system is determined by approximately the size of the outermost smallest zone width, and thus, is tightly connected to advancements in the lithographic fabrication process of zone plates, currently allowing hard X-ray microscopy resolutions well below 50 nm.

Whereas SR-based zone-plate TXM setups are frequently used in 2D, as well as in 3D when combined with Rapamycin a rotation stage for tomography, it was not until recently that a first desktop TXM CT system was implemented [21], which is operated with a commercial X-ray tube. An initial TXM CT measurement performed on this system provided a 3D reconstruction of an osteocyte lacunae and radiating canaliculi of a tibial trabecula in the mouse [22]. Although the spatial resolution of the system in 2D has been reported to be below 50 nm [22], canaliculi in the 3D reconstructions were interrupted. Therefore, further

refinements to this technology are needed in order to accurately model the canaliculi in 3D. Higher GDC-0199 ic50 spatial resolutions can be achieved using electrons instead of X-rays, where the resolution of an electron microscope increases in a manner that is inversely proportional to the square root of the applied voltage, and is typically in the nanometer range. TEM has been extensively used to investigate in 2D the ultrastructure of osteoblasts and osteocytes including their dendritic processes.

The morphology of osteocytes and their processes were further characterized in 3D by successive serial sectioning and TEM imaging [23]. More recently, Kamioka et al. adopted TEM computed tomography (TEM CT) on an ultra-high voltage electron microscope, where silver-stained osteocytes in 3-μm chick calvaria sections were Interleukin-3 receptor assessed at an accelerating voltage of 2 MeV and at a nominal resolution of 16 nm [24]. Prominent silver deposition for young osteocytes, which has been observed in their nuclei and in the pericellular space, was used to segment the cell nuclei, cell bodies, and the osteocyte processes (Fig. 1B). Kamioka and colleagues found that the surface of the osteocyte network was irregular and that the size and shape of the cell processes varied significantly. Besides the demanding sample preparation, a major problem of TEM is the fact that for a dense material like bone, even at ultra-high voltages, the maximal sample thickness that can be penetrated by electrons is only a few μm due to strong scattering and absorption for thicker specimens.

Osteochondral transplantation is a possible option for these youn

Osteochondral transplantation is a possible option for these younger, more active patients but access to fresh

tissue is extremely difficult. Fresh osteochondral transplants provide Selleckchem ZVADFMK the best case scenario for cell viability and matrix integrity but fresh transplant is fraught with technical difficulties. This tissue should be harvested within 24 h of death of the donor and is typically transplanted within 48–72 h after harvest [37]. This time frame is too short to perform the extensive testing required to rule out the possibility of transmission of infectious diseases. Considering that joint injury and osteoarthritis are not life threatening, the risk may not be warranted. Another significant technical difficulty is that matching for size and contour, which are important factors for long term successful outcomes, is extremely difficult on such short notice [19]. Making arrangements for complicated joint replacement surgery on short notice can result in logistic problems in arranging the operating room, appropriate surgical staff, surgeon and even the patient. Currently, blood/HLA typing is not performed as articular cartilage is considered immune privileged. That said, the cartilage is transplanted on bone and there can be minor immune reaction to the transplanted

bone. This is typically self-limited as the transplant bone is replaced with host bone if only a small amount is transplanted. In the future, blood/HLA typing may be employed to limit the immune reaction which adds another layer of complexity to performing this surgery on short notice. To address these issues, hypothermic storage at 4 °C for UK-371804 in vivo a limited time (28–42 days) is used to increase selleck screening library the supply [41] and [110]. Unfortunately, tissue deterioration begins after only 7–14 days [65]. The lack of normal mechanical stimulation impairs the efficiency of nutrient and waste transport, and decreases cytokine secretion (IL-1 and TNF-α) as reviewed by Kim, Teng and Dang [58].

The ability to store articular cartilage indefinitely would allow for precise size/contour matching, pre-surgical planning, testing for infectious diseases, possible blood typing and appropriate surgical timing for the patient, operating staff and surgeon. Successful cryopreservation of articular cartilage, by either classical methods or vitrification, can extend the availability of the tissue and allow long-term banking of articular cartilage. Successful cryopreservation and banking of articular cartilage will enable easier and more efficient utilization of straightforward protocols for transplantation. From a cryopreservation perspective, articular cartilage with its extracellular matrix containing no lymphatic, nervous or vascular systems and only one cell type is considered to be a stepping stone for the transition from simple cell to complex tissue cryopreservation with high cell viability and function.

Plasma P increased somewhat in S2* but little change was seen in

Plasma P increased somewhat in S2* but little change was seen in S5*. Problems with treatment compliance were acknowledged and the therapy was subsequently terminated. Candidate gene analysis

of the SLC34AC gene, encoding a type IIc Cyclopamine sodium-phosphate transporter (NaPi-IIc) expressed in the kidney, was conducted in the DNA sample from the youngest affected sibling (S1*) (Fig. 1). Three SNPs were found for which the other family members were then screened. Two of the SNPs had been previously reported on the NCBI database and had been assigned the following reference numbers rs28542318 and rs74842953. These SNPs referred to a non-synonymous mutation E513V (c.1538A > T) and a synonymous mutation L599L (c.1795 T > C) respectively. In silico mutation analysis suggested that these two SNPs were benign polymorphisms and they were unique to S1* and were not present in the other investigated family members. The third SNP was a novel non-synonymous mutation resulting in an amino acid change S168F (c.503C > T). Blast alignment results indicated that the flanking amino acid regions at the site of the S168F mutation were highly conserved throughout species

( Table 3). In silico mutation analysis predicted that this SNP would cause a damaging RO4929097 solubility dmso mutation in the NaPi-IIc protein, the likely cause being the loss of function of the transmembrane domain spanning 133–188 amino acids. Prediction analysis, suggested that the protein product containing S168F was of normal length (599 amino acids) and that the correct reading frame was maintained. The S168F mutation was present homozygously in the affected siblings (S5*, S2* and S1*) and heterozygously in the unaffected family members ( Fig. 1). The Republic of The Gambia (latitude 13°N) in Adenosine West Africa has a hot and dry tropical climate with a single wet season from June to October. There is abundant UVB-containing sunshine throughout the year and a lifestyle that does not limit skin UVB-exposure. Cases of rickets have, however, been reported and have been attributed predominantly

to a chronically low dietary calcium (Ca) intake leading to a 1,25(OH)2D-driven increase in FGF23 leading to urinary phosphate (P) wasting and rickets [1] and [2]. The family described in this study is, however, strikingly different to our previous reports of rickets in The Gambia. To our knowledge, this study documents the first cases of hereditary hypophosphataemic rickets with hypercalciuria (HHRH) in Africa. The cause of HHRH is a mutation within the gene encoding the Type IIc sodium-phosphate co-transporter [3] and [4]. There are two major NaPi co-transporters involved in P reabsorption in the proximal tubule of the kidney. These are NaPi-Type IIa and -Type IIc, both of which are regulated by FGF23 and PTH.

Mulder et al3 and Ishioka et al4 initially described diverticulot

Mulder et al3 and Ishioka et al4 initially described diverticulotomy by using freehand endoscope manipulation. Sakai et al5 demonstrated that using a cap at the tip of the endoscope offers 3-MA datasheet better visualization of the septum. Evrard et al6 showed better exposition of the septum with the use of a soft diverticuloscope. The fear of bleeding during treatment prompted Mulder et al3 and 7 to use argon plasma coagulation (APC) instead of conventional current, requiring multiple sessions with the risk of inducing fibrosis.

The availability of a soft, plastic diverticuloscope that mimics the Van Overbeek diverticuloscope8 has allowed better septum exposure and endoscope stability without the risk of trauma associated with the use of the rigid instrument. Moreover, it allows an extended section of the septum and protects the airway from aspiration in nonintubated patients. A previous study showed that diverticulotomy with a flexible endoscope and soft diverticuloscope is an effective treatment for ZD,6 and another suggested that this treatment was safer and more effective than freehand

treatment.9 This initial experience RO4929097 clinical trial prompted us to modify the technique with systematic clipping of the bottom section at the end of the procedure to reduce the risk of perforation and improve hemostasis. We report our long-term results of ZD treatment by using flexible endoscopy assisted by the use of a soft diverticuloscope. This study was conducted in accordance with the ethical principles of the Declaration of Helsinki, in compliance with good clinical

practice and according to local regulations. This work was not supported financially or otherwise by any external sources. All patients gave informed consent after explanation of the technique. Ethical approval for the study was obtained from the Institutional Review Board from our center, reference P2010/353. All patients with ZD who were treated in our medical-surgical department between July 2002 and June 2011 were included in the study. None of them Liothyronine Sodium were included in our previous report.6 Files were reviewed retrospectively, and clinical data were recorded (demographics, symptoms, dysphagia score, endoscopic treatment technique, adverse events, and outcome). Adverse events are defined by the Cotton et al10 severity scoring system. Patients were asked to fill in a questionnaire to describe their symptoms and quantify dysphagia before and after treatment. Dysphagia scores 1 month after treatment were available only for patients who were seen in the outpatient clinic at that time. Because a significant number of patients came from abroad, early follow-up in them was performed by the referring physician, and results were not available at the time of data collection. The Dakkak and Bennett11 score of dysphagia (score 0, no dysphagia; score 1, dysphagia to solids; score 2, dysphagia to semi-solids; score 3, dysphagia to liquids; score 4, aphagia) was used to quantify dysphagia.

1A) These 31 sites represented our best judgment of conditions b

1A). These 31 sites represented our best judgment of conditions before the BMN 673 mouse oil entered the estuaries. We were prevented from accessing most marshes until the fall 2010. Various agency and satellite image analyses at that time indicated that the most prominent oiling was in east and west Barataria Bay and eastern Terrebonne Bay. We focused on these three areas and chose the target areas before the field trip began, and then made our final selection while in the field and before landing the boat. Subsequent sampling included these three general areas, but the same exact sites were not always re-sampled because of landowner

permission, erosion, or logistical issues (principally the shallow water depth that hindered Neratinib clinical trial boat access). A core set of 12–13 site locations were sampled on each trip. Thirty sites were established on the northern edge of Bay Batiste in February 2011 (Fig. 1C). These were clusters of 3 stations 10 m apart and are the same sites used by McClenachan et al. (2013) for a marsh erosion study. Sites were marked

with a plastic 0.25 m2 quadrat to facilitate repeated sampling at the same location. We had no access to data on oil concentration to assist in site selection for any site until late summer 2011. We collected 405 surface-sediment samples from Louisiana coastal wetlands during May 2010 (n = 31), September 2010 (n = 64), February 2011 (n = 30), May 2011 (n = 87), September 2011 (n = 66), June 2012 (n = 22), August 2012 (n = 30), September 2012 (n = 30), October 2012 (n = 15), and June 2013 (n = 30) ( Fig. 1). The majority of the samples were collected within 10 m of the shoreline. Others were collected every 20 m along eight 90 m transects in June 2011, and five 100 m transects Tacrolimus (FK506) in September 2011. These transects were perpendicular to the wetland/water interface. Sampling in February 2011, August 2012, September 2012, and June 2013 were within 1 m of each other. The primary emergent vegetation was Spartina alterniflora

and Juncus sp. with minor amounts of Schneoplectus americanus. The wetland type is commonly known as a ‘salt marsh’. All sediment samples were collected as a composite sample of the upper 5 cm, stored on ice until delivery to the laboratory, and either immediately extracted or refrigerated at 4 °C for no more than 14 days until extraction, as recommended by the US EPA (2007). The samples were analyzed using GC/MS-SIM that targeted 28 alkanes, 18 parent PAHs, and 25 alkyl homolog groups (Table 2). The target petrogenic compounds were extracted from the sediment samples using EPA SW-846 method 3540C (US EPA, 2000). Reagent grade or pesticide grade solvents were used in all the extractions and analyses. Samples were homogenized and a 15–20 g subsample was weighed, spiked with surrogate recovery standards (5-alpha androstane and phenanthrene-d10, AccuStandard, Inc.

Differences in grammatical or lexical class may not, however, be

Differences in grammatical or lexical class may not, however, be the principle factor in the neural differentiation between nouns and verbs. As one variable of interest, word meaning, or semantics, has frequently been discussed as an underlying determinant of noun/verb dissociations (Pulvermüller, Lutzenberger et al., 1999, Shallice, 1988, Vigliocco et al., 2011 and Warrington and Shallice, 1984). An essential confound exists in the literature as most verbs are undeniably words used to speak about actions whereas most nouns refer to objects, so it is hardly possible to match and control for relevant semantic differences between check details the lexical classes; furthermore, were one to succeed in precisely

matching sets of nouns and verbs for factors such as the concreteness of their object reference and intensity of their action relationship, one might, from a linguistic perspective, still argue that such selections would certainly be far from representing typical specimens from the lexical groups.

Given this seemingly hopeless confound of lexical Cilengitide order class with semantics, it is therefore unsurprising that many scholars have tried to trace the “lexical” differences to their semantic origins, at least as far as putative word class specific brain activation patterns are concerned. Ingenious attempts have been made to clarify this issue by varying semantic properties within the lexical classes so that consistent noun/verb differences in brain activation – for example in the middle-temporal cortex (Bedny et al. 2008) – reveal more genuine lexical class differences. In addition, many authors have attempted to strip words of their semantics by contrasting homophonous pseudowords in noun and verb context (to wug vs. the wug), thus providing a tool for ascertaining differential representation

of lexical categories (Cappelletti et al., 2008, Laiacona and Caramazza, 2004, Shapiro and Caramazza, 2003, Shapiro et al., 2006 and Shapiro et al., 2001). However, taking on the role of an advocatus diaboli, one might still argue that the phrase “to wug” suggests an action (e.g., whacking) whereas the context “the wug” is more compatible with an object (a rug) interpretation and, therefore, these pseudowords were not truly stripped of semantic associations, but were, in fact, semantically biased by Pyruvate dehydrogenase lipoamide kinase isozyme 1 the contexts in which they were presented: as the authors did not explore this possibility empirically, this interpretation (which has earlier been suggested and supported by Pulvermüller, Kherif, Hauk, Mohr, and Nimmo-Smith (2009) and Vigliocco et al. (2011)) cannot be ruled out at this point. Further evidence for representation of lexical categories in the brain comes from differential brain activity in response to homophonous noun and verb affixes presented in noun and verb contexts ( Pulvermüller & Shtyrov, 2009), which persist even after the contributions of the noun/verb stems are subtracted.

Assuming a two state model, the observed mean-residue ellipticity

Assuming a two state model, the observed mean-residue ellipticity at 222 nm ([Θ]obs222) was converted into α-helix fraction (fH) using the method proposed

by Rohl and Baldwin (1998) and previously described ( Konno et al., 2001). The lipid bilayers were obtained from giant unilamellar vesicles (GUVs), which were positioned onto the chip MK-1775 aperture by application of negative pressure. The GUVs burst as soon as they touch the glass surface of the chip and form a bilayer that spans the aperture (Sondermann et al., 2006). Asolectin (Sigma), a negatively charged mixture of lipids, was used to form artificial membranes. GUVs were formed by electroswelling, using the Nanion Technologies (Munich, Germany) device Vesicle Prep Pro©. 20 μL of 10 mg/mL lipid solution (in chloroform) were deposited onto an indium tin oxide (ITO) coated glass plate and evaporated for 45–60 min. A nitrile ring was placed around the dried lipid film and filled with 350 μL of 250 mM D-Sorbitol dissolved in Milli-Q water. A second ITO coated glass plate was placed on top of the ring. An AC voltage of 3 V peak-to-peak Stem Cell Compound Library molecular weight amplitude at 5 Hz frequency was supplied to the ITO slides over a

period of 2 h at 36 °C (modified from Sondermann et al., 2006). The formed vesicles were kept in plastic vials under refrigeration (4 °C) until use or used immediately. GUVs suspensions were always observed under light microscope prior to use. The experiments were performed with the automated Patch-Clamp device Port-a-Patch (Nanion Technologies – Munich, Germany), using borosilicate glass chips NPC-1 with aperture

diameter of approximately 1 μm. The resistance of the apertures was approximately 1–3 MΩ in 150 mM HCl solution. Current signals were amplified and recorded by an amplifier EPC-10 (Heka Elektronik, Lambrecht, Etoposide supplier Germany) and an analogical/digital interface ITC-1600. The system was computer controlled by the PatchControl™ software (Nanion) (Fertig et al., 2002 and Sondermann et al., 2006). During the experiments symmetrical solution of 150 mM HCl with 5 mM Tris was used. After a seal was formed (Rm > 500 mΩ), the peptides diluted with Milli-Q water at a 5 μM concentration were added to the cis side of the chip (top) to observe the single channel activity. The volume of peptide solution was never superior to 10% of the solution at the cis side. Voltage pulses were applied at the trans side of the chip (bottom). Usually, single channel activity started approximately 10 min after adding the peptides, as monitored by a constant Vhold of −100 mV. Single channel conductance of incorporated channels was determined under positive and negative voltage pulses. The experiments were performed at room temperature (∼22 °C). The data was analyzed by PatchMaster and Matlab softwares.

Table 2 shows the effects

of juglone on

Table 2 shows the effects

of juglone on Anticancer Compound Library chemical structure the ADP/O and respiratory control ratios (RC). As noted, juglone reduced significantly the ADP/O ratio already at the concentration of 1 μM when β-hydroxybutyrate was the substrate. At the concentration of 5 μM the ADP/O ratio could no longer be determined. The respiratory control ratio was also reduced and eventually abolished, depending on the concentration. Similar results were obtained when succinate was the substrate, but at somewhat higher concentrations. The uncoupling action of juglone was further investigated by measuring the ATPase, NADH-oxidase and succinate-oxidase activities of rat liver mitochondria. The ATPase activity was measured using mitochondria under three different conditions: intact (coupled), freeze-thawing disrupted and 2,4-dinitrophenol uncoupled. Fig. 8A shows that the ATPase activity was stimulated by juglone in the range between 1 and 10 μM, but with a maximum at 2.5 μM. The ATPase activity of disrupted and uncoupled mitochondria, however, was relatively insensitive to juglone in the range up to 2.5 or 5 μM, and inhibited at higher concentrations. The selleck chemical actions of juglone on the NADH- and succinate-oxidase activities are shown in Fig. 8B. The NADH-oxidase activity was stimulated at concentrations between 5 and 10 μM; the succinate-oxidase activity,

however, was not significantly affected. The main conclusion that can be drawn from the bulk of the data obtained in the present work is that juglone is active on liver metabolism and able to affect several metabolic routes which are linked in some way to energy oxyclozanide metabolism. In general, most observations in the perfused liver are compatible with its reported uncoupling action. The most important observations, which have also been traditionally reported for other uncouplers of oxidative phosphorylation are: a)

stimulation of oxygen consumption at low concentrations (Soboll et al., 1978 and Suzuki-Kemmelmeier and Bracht, 1989); b) diminution of the ATP content combined with diminutions in the ATP/ADP and ATP/AMP ratios (Soboll et al., 1978); c) increase in the NADH/NAD+ ratio (Soboll et al., 1978 and Suzuki-Kemmelmeier and Bracht, 1989); d) inhibition of gluconeogenesis (Kelmer-Bracht and Bracht, 1993 and Suzuki-Kemmelmeier and Bracht, 1989) from two different substrates, namely lactate and alanine; e) stimulation of glycolysis as a cytosolic compensatory phenomenon for the diminished mitochondrial ATP production (Soboll et al., 1978 and Suzuki-Kemmelmeier and Bracht, 1989); f) stimulation of glycogenolysis as a means of providing glucose 6-phosphate for the increased glycolytic flux (Lopez et al., 1998 and Soboll et al., 1978). Experiments with isolated mitochondria, based on the original observations of Makawiti et al. (1990), allowed to characterize further the actions of juglone on the organelle.

The authors effectively balance between these two endpoints of hi

The authors effectively balance between these two endpoints of historical ignorance. The text conveys a great deal of information, but is quite accessible to a non-specialist reader interested in natural history and environmental change. The scholarship is thorough, balanced, and impeccable, and the writing is engaging. The text is nicely illustrated with diagrams, historic maps, and matched

historic and contemporary photographs. The matched photographs are particularly effective because juxtaposed on the same page, facilitating visual comparison of changes through time. The title refers to irreversible changes to the river through the Tucson Basin, mainly from urbanization and groundwater overdrafts. The authors conclude the book by noting that, although “the Santa Cruz River of old can be neither NLG919 restored nor revived” (p. 182), the river can be managed to minimize flood risk and maximize ecosystem services. This “will require both an acknowledgement ZD1839 ic50 of history and fresh perspectives on how to manage rivers and floodplains in urban areas of the Southwest” (p. 182). This

book provides a firm foundation for such a path forward. “
“Lagoons are widely distributed throughout the world ocean coasts. They constitute about 13 percent of the total world coastline (Barnes, 1980). They represent 5.3 percent of European coastlines (Razinkovas et al., 2008), with more than 600 lagoons in the Mediterranean area alone (Gaertner-Mazouni and De Wit, 2012). From geological and geomorphological viewpoints, coastal lagoons are ephemeral systems that can change in time (becoming estuaries or infilled; Davies, 1980). The nature of this change depends on the main factors controlling their evolution, such as mean sea level, hydrodynamic setting, river sediment supply and pre-existing topography. As observed by Duck and da Silva (2012), however, these coastal forms are seldom if ever allowed to evolve naturally. They are often modified by Racecadotril human intervention typically

to improve navigability or in attempts to maintain the environmental status quo. By controlling their depth and topography, humans have exploited them for many centuries for food production (fisheries, gathering of plants and algae, salt extraction, aquaculture, etc.) (Chapman, 2012). These modifications can transform radically the lagoon ecosystem. Human activities have also influenced the evolution of the Lagoon of Venice (Italy) over the centuries (Gatto and Carbognin, 1981, Favero, 1985, Carbognin, 1992, Ravera, 2000, Brambati et al., 2003 and Tosi et al., 2009). Together with the historical city of Venice, the Venice Lagoon is a UNESCO World Cultural and Natural Heritage Site. The first human remains in the lagoon area date back to the upper Paleolithic age (50,000–10,000 BC). The lithic remains found in Altino (Fig.

This cycle was repeated until the enzymatic activity become null

This cycle was repeated until the enzymatic activity become null. The influences of pH and temperature on β-glucosidase activities were determined using the standard assay for the free and immobilised enzymes, except that the pH values were modified to a range of 2.0–8.0 (Mcllvaine, 1921) and the temperature values BMS-387032 mouse ranged from 10 to 60 °C. The pH stability

of β-glucosidase was determined by incubating the free enzyme solution or the alginate beads in the pH range of 2.0–8.0 for 30 min, on ice. After incubation, the mixture was used for determining residual activity, according to standard assay, using pNPβGlc as the substrate. Thermal stability was investigated by incubating the enzymatic solution or the alginate beads in 50 mM sodium phosphate buffer, pH 6.0 or 5.5, respectively, at temperatures of 45 and 50 °C for different times. After pre-incubation, aliquots of the enzymes or 4 alginate beads were collected and submitted to the standard MLN0128 assay, measuring the remaining activity. The relative activities were calculated in

relation to β-glucosidase activity without pre-incubation, which was considered to be 100%. Results of the analyses are presented as mean ± SD for three measurements. The Michaelis–Menten constant (KM) and Vmax for substrate hydrolysis by the free enzyme and the KMapp value for the immobilised enzyme were calculated by the Michaelis–Menten plot. Concentrations of pNPβGlc varied from 0.2 to 5.0 mM. The inhibition Cytidine deaminase constant (Ki) for the free enzyme using glucose as inhibitor was determined by varying the pNPβGlc concentrations from 0.05 to 1.2 mM in the presence of 50, 100 or 120 mM of glucose. Enzymatic assays were performed with various synthetic, natural and polymeric substrates. The reaction mixtures contained 650 μL of 50 mM sodium phosphate buffer pH 6.0, 0–100 μL of enzyme solution and 250 μL of synthetic substrates (0.5 mM)

or celobiose, lactose, maltose, gentiobiose, melibiose and sucrose (2.5 mM) or cellulose (0.025%). Activities were measured under standard assay conditions at 40 °C. The data presented for all enzyme activity determinations are mean values ± SD of three measurements. The effects of ions, simple sugars and reducing agents on enzyme activity were assayed by the standard methods. Reaction mixtures contained 450 μL of 50 mM sodium phosphate buffer pH 6.0, 0–100 μL of the enzyme solution and 200 μL of the compounds (0.2 and 2 mM). The data presented for all enzyme activity assays are mean values ± SD of measurements performed in triplicate. The soy molasses samples were kindly donated by Melaços Brasileiros Ltda., Saltinho, São Paulo, Brazil. One gram samples of soy molasses were incubated with either 10 U of free β-glucosidase in 50 mM sodium phosphate buffer pH 6.0 (10 mL) or with a calculated number of beads corresponding to 10 U of β-glucosidase in 50 mM sodium phosphate buffer pH 5.