The understanding of the underpinnings of such interval CRCs is of importance because it may permit identification of modifiable factors, for example gaps in knowledge and training on the recognition of nonpolypoid neoplasms and their endoscopic resection. In this case, tailored educational programs would improve the awareness and help to shape practical skills, to ultimately safeguard the quality of colonoscopy. Furthermore, it is important to understand whether

certain molecular features of the inflamed mucosa could augment the risk of cancer progression. Such information may help to develop personalized (ie, molecular-based) surveillance strategies. Two recent studies exploring the cause of sporadic interval CRCs in the general population found missed lesions represent by far the most important contributor (>50% of all interval CRCs).22 and 23 ABT-199 cell line VX-809 price Undoubtedly, missed lesions are likely to account for a significant proportion of interval CRCs in IBD, although a thorough analysis using structured algorithms24 has not yet been performed. A recent population-based analysis by Wang and colleagues,25 using SEER

cancer registry data from 55,008 older patients with CRC, found rates of early/missed CRCs were three-fold greater in IBD than in patients without IBD (15.1% for Crohn’s disease, 15.8% for UC vs 5.8% for patients without IBD; P<.001). Early/missed CRCs were defined as CRCs identified within 6 to 36 months after a colonoscopic examination that did not detect cancer. This study was based on administrative data, and therefore lacked detail about the completeness of colonoscopy, bowel preparation, extent of colitis, characteristics of mucosal lesions identified at the baseline examination, and resection outcomes. Such observations underscore the importance of meticulous inspection of the entire colonic mucosa, which should be ideally clean and free of inflammation, and the need for formal training of

the endoscopist in the recognition of IBD neoplasms. Presence of active Phosphatidylinositol diacylglycerol-lyase or chronic background inflammation and the diversity in endoscopic appearance of dysplasia by IBD may, however, increase the complexity of diagnosis. Fig. 1 illustrates a lateral spreading tumor of granular subtype, which could have been missed at a previous examination. A substantial number of studies demonstrated that indigo carmine– or methylene blue–guided chromoendoscopy (CE) improves the diagnostic yield of dysplasia and invasive CRC during IBD surveillance. This is not surprising, because a significant proportion26, 27 and 28 of dysplastic lesions in patients with IBD appear to have a flat appearance, as illustrated in Table 1. Pancolonic CE delineates the borders and permits a detailed analysis of the epithelial surface, thus facilitating the diagnosis of subtle lesions and their endoscopic resection.

To investigate the correlation between the data that can be obtained using the classical kOPA test and the newly developed fOPA method, we measured fOPA titers in a panel of sera displaying a wide range

of kOPA titers to GBS Ia. Remarkably, a good correlation (R2 = 0.82, p < 0.05) between fOPA and kOPA read outs was observed (Fig. 8). PD0325901 molecular weight A subset of sera was also tested against GBS serotype III using the isolate COH1 and a good correlation between the two methods (R2 = 0.85, p < 0.05) was obtained also in this case (data not shown). The data indicate that the fOPA method can be used to test functional antibodies against different serotypes. We developed an opsonophagocytosis assay for GBS using pHrodo™ labeled bacteria. Our method offers several advantages over both killing-based and other fluorescence-based opsonophagocytic assays. The most commonly-used fluorophores in OPA assays are fluorescein (fluorescein, dicarboxyfluorescein, oregon green, dihydrodichlorofluorescein) or Alexa Fluor derivatives. Flow cytometry based on those fluorophores can detect cell-associated fluorescence but cannot distinguish between internalized and adhering bacteria, necessitating quenching steps with trypan blue or signaling pathway ethidium bromide to clean out the background fluorescence of externally bound bacteria.

The pHrodo™-based assay provides sensitive detection without the need for quenching or washings steps, saving time and eliminating measurement uncertainty. Indeed, pHrodo™ is a pH sensitive fluorophore showing a very low fluorescent signal at the neutral pH of extracellular and cytoplasmic environment and a bright fluorescent signal in acidic compartments, such as phago-lysosomes, deriving from check details the fusion of phagosome-containing bacteria with lysosomes which occurs immediately after internalization. As shown by confocal microscopy images, GBS bacteria labeled with pHrodo™ exhibit low fluorescence outside the cell, yet emit a bright

red fluorescence after internalization into the acidic environment of the phagocyte. By determining whether phagosome containing bacteria mature to phago-lysosome acidic compartments, the pHrodo™ assay is predictive of phagocytic killing. Several different mechanisms can lead to bacterial survival after phagocytosis, rendering the phagocytosis measurement non strictly indicative of pathogen clearance. For instance, it has been observed that certain mycobacteria (e.g. Mycobacterium avium, Mycobacterium tubercolosis) are not always killed even when enclosed in phagocytic cells, because the phagosome-lysosome fusion is not accompanied by the normal acidification that creates the appropriate conditions for killing ( Hornef et al., 2002, Bellaire et al., 2005 and Huynh and Grinstein, 2007). Further, the phagosome-lysosome fusion may not occur or the phagosome may not close.

Factor Inhibiting HIF (FIH)

is a 2OG oxygenase that catalyzes the hydroxylation of an asparagine residue within the C-terminal transactivation domain of HIF-α, thereby inhibiting the binding of co-activators CREB-binding protein (CBP) and p300 to the HIF transcriptional complex. Conversely, FIH inactivation facilitates CBP/p300 recruitment and results in increased HIF target gene expression under hypoxia.86 In the kidney, FIH has been detected in REPC, podocytes and in the distal tubule.[90] and [93] While the role of PHDs and FIH in the regulation of HIF activity is well established, alternative hydroxylation targets have been identified Y-27632 purchase and are likely to impact hypoxia and EPO responses in the kidney.[85], [94] and [95] Furthermore, Selleckchem Etoposide it is likely that renal EPO synthesis is modulated by epigenetic changes

that are carried out by non-HIF 2OG oxygenases. Although nothing is known about their role in renal physiology, 2OG oxygenases, which contain a jumonji domain, catalyze the demethylation of methylated histones,85 and are likely to provide additional functional links between alterations in renal pO2 levels and gene expression.96 Although in vitro approaches identified HIF-1 as the transcription factor responsible for the hypoxic induction of EPO, 97 HIF-2 has now emerged as the main regulator of EPO production in vivo ( Fig. 2). Several lines of evidence exist that support this notion: a) the location of HIF-2α-expressing renal interstitial

cells coincides with the location of REPC [12] and [98]; b) genetic studies in mice have demonstrated that renal and liver EPO synthesis is HIF-2- and not HIF-1-dependent, as did siRNA and chromatin immunoprecipitation (ChIP)-based studies in certain EPO-producing cell lines [72], [99] and [100]; c) genetic analysis of patients with inherited forms of erythrocytosis have revealed mutations in HIF2Α but not in HIF1Α (see section on Reverse transcriptase HIF pathway mutations in patients with secondary erythrocytosis); and d) genetic variants of HIF2A have been associated with high altitude dwellers who are protected from chronic mountain sickness (see section on molecular adaptation to life at high altitude). While HIF-1α is ubiquitously expressed, HIF-2α expression is more restricted. HIF-2α was initially identified in endothelial cells, subsequent studies however demonstrated expression in hepatocytes, cardiomyocytes, glial cells, type-II pneumocytes, and in renal peritubular interstitial cells.[98] and [101] The analysis of HIF-1α and HIF-2α knockout mice provided the first major insights into the functional differences between these two HIF homologs.

For the separation of sucrose, the zeolites CaX and MgX presented the most promising results, since they adsorbed about 250 g/L after 60 min of reaction. The amount of glucose adsorbed after 60 min increased according to the following

sequence of zeolite forms: Ba2+ < Mg2+ < Ca2+ < K+ < Sr2 < Na+. Considering the fructose separation, the amount adsorbed increased according to the following sequence of zeolite forms: Sr2 < Ca2+ < K+ < Mg2+ < Ba2+ < Na+. Considering the sucrose separation, the amount adsorbed increased according to the following sequence of zeolite forms: K+ < Na+ < Sr2 < Ba2+ < Ca2+ < Mg2+. Apoptosis Compound Library high throughput Heper et al. (2007) evaluated the separation of glucose and fructose using the Y zeolite. Considering the fructose adsorption, the amount adsorbed increased

according to the sequence NH4+ < Mg2+ < Na+ < Ca2+, while the amount of glucose adsorbed increased according to the sequence NH4+ < Mg2+ < Ca2+ < Na+. These results are similar to the one obtained in this work. Gramblicka & Polakovic (2007) reported the capacity of the adsorbents Diaion, Dowex, Lewatit and Amberlite to recovery of individual saccharides and verified that the adsorbed amounts decreased in the order fructose > glucose > sucrose > kestose > nystose > fructofuranosylnystose. In addition, Gramblicka & Polakovic (2007) verified that the sieve effect of the resins were the primary cause of the Protease Inhibitor Library order different partitioning of the investigated saccharides between the solid and liquid phases. This also explains that no effect of the concentration on the distribution coefficients was observed at the multicomponent adsorption from the mixture of FOS. The authors also O-methylated flavonoid assumed

that obtained isotherms for individual FOS were not affected by the presence of other species of the mixture. The low performance of the BaX zeolite to recovery glucose could be due to the fact that the hydrated Ba ions cannot migrate into the sodalite unit and the hexagonal prism during ion exchange because of their large ionic radii. They occupy positions in the supercage and can interact with the adsorbents even at a low degree of exchange (Schöllner, Einicke, & Gläser, 1993). Based on the experimental results, the most appropriated forms to separate glucose, fructose and sucrose from the reaction medium are the forms NaX, NaX or BaX and MgX or CaX, respectively. Nevertheless, the choice of the most appropriated form to separate these sugars can be made according to a numerical analysis of the model parameters in terms of adsorption rates and mass transfer resistances involved in the process. In this sense, therefore, the experimental data from Fig. 1 were used to estimate the model parameters for each zeolite form, which are presented at Table 1. Before the analysis of the model parameters, some aspects concerning the convergence and stability of the parameter estimation should be overviewed.

Also, inside the RV there were placed a few (5–7) 2 mm diameter glass beads that helped to damp liquid motion when pyruvic acid first entered. The pH of the injected substrate was automatically measured during the dissolution procedure using a pH electrode (ASP200-2-1M-BNC, Active robots Ltd., Radstock, UK) placed in the RV. The pH electrode was connected to a custom built amplifier that had a variable output voltage in response Obeticholic Acid to changes in pH, see Supplementary

information. The amplifier was connected to an analogue to digital converter input on the microcontroller. By titrating 45 mg pyruvic acid against 2.0 M sodium hydroxide over a pH range of 1.8-–13.0, voltage versus pH was plotted and used to generate a linear calibration equation. The pH electrode could also be calibrated from within the Arduino software by measuring the electrode voltage in 3 different buffer solutions (pH 4, pH 7, pH 10) and calculating a linear

equation for pH versus voltage. Also connected into the RV was a 6 mm O.D. pipe connected to a vacuum pump (GAST GF3, Gast Manufacturing Inc., MI) to reduce back pressure during transfer of hyperpolarized solution. The vacuum pump was gated on/off by the HyperSense DNP polarizer. Injection volumes for each species are limited to ensure that the circulation INCB024360 of the animals is not overloaded. The physical constraints of an MRI scanner require a long length of cannula line for i.v. injections, resulting in a significant dead volume that contributes

to the injection volume. This is problematic where hyperpolarized signal is limited. If, for example, saline occupies the dead volume of the cannula then, during injection, its volume must be considered part of the dose and yet it does Staurosporine nmr not contribute to the measured hyperpolarized signal. To increase the percentage the hyperpolarized compound contributes to the injected volume, the dead volume was reduced by splitting the cannula into two pathways without introducing additional dead volume; one pathway was then used as a waste stream for clearing the dead space volume whilst the other was used for drug administration into the animal. Flow direction was computer-controlled by valves. A fluid diverter cannula was constructed using two types of tubing: 0.96 mm O.D. polyethylene tube (Portex, Smiths Medical, St. Paul, MN), hereby referred to as ‘small tube’ and 1.0 mm I.D. Tygon tube (Cole-Parmer, London, UK), hereby referred to as ‘large tube’. Tygon tubing was used as its mechanical properties permit multiple compressions without permanent damage. A 19 gauge Luer hub was drilled to enlarge its inner diameter to 2 mm, see Fig. 3, into which the ends of three 30 mm lengths of small tubing were inserted to ensure the hole was almost completely occupied; one was used for the waste pathway, one for the animal pathway which was inserted into a rat vein, whilst the third one was unused and blanked off. The tubes were then sealed to the Luer hub with glue.

Several enzymes are sensitive to inhibition by high ionic strengths and altering the concentrations of charged substrates and the pH of the buffer may also affect this. The ionic strength of assay media is seldom stated, although this can be calculated if the full composition and pH of the assay mixture is given, it would be helpful if all authors were required to state the value. Other additives such as chelating or reducing compounds, which are needed for the

this website activity of some enzymes, will inhibit others and specific metabolites are required to activate some enzymes, such as acetyl Co-A for pyruvate carboxylase (EC 6.4.1.1) and N-acetyl-l-glutamate for carbamoyl-phosphate synthase (ammonia) (EC 6.3.4.16). Various attempts have been made to define assay media that are appropriate for determining the behaviour of enzymes under “in vivo-like” conditions ( van Eunen et al., 2010 and Goel et al., 2012). However, from the above examples, it should be clear that it is unlikely that a universal buffer medium, suitable for all enzymes

in all tissues and organelles, will be found. Indeed different Entinostat cell line conditions should apply to the same enzymes from different sources. Individual standards will be required for each organism, organ and organelle to be studied, bearing in mind that these may not be constant under all metabolic conditions. Perhaps the answer will lie in more complex mixtures, including proteins as buffers, that more closely mimic the, crowded, in vivo environments of groups of enzymes. In its attempts at formulating more physiologically relevant assay conditions the STRENDA Commission needs advice from those working with specific systems. None of the authors have any conflict of interest. “
“Due to a production error, the issue 16P3 starts with page 1 instead of page 209 as a continuation of 16P2. The Publisher sincerely apologizes to the readers and deeply regrets any inconvenience caused. “
“Foreword v Preface vii Acknowledgements xi

Biographies xiii 1. Vaccine evolution 1 Appendices I Glossary XI Disclaimers XXIII Copyrights permission texts for non-original illustrations XXVII Index XXXVII Supplementary Data XLV “
“The history of infectious disease Aurora Kinase shows unequivocally that vaccination is the cheapest and most effective form of medical intervention ever devised. Application of the original strategy, developed (in 1796) when Edward Jenner scarified pustular material recovered from the teat of an infected cow into the arm of a young boy, James Phipps, then challenged him later with virulent smallpox virus, led to the global elimination of that terrible disease some 200 years later. Though we may still lack optimal vaccines, the toll of catastrophic infections like cholera was substantially blunted through the 19th century by cleaning up the water supply.

In Fig. 3, crossings during the readout were seen in the linear-order phases in the bipolar sequence. This is characteristic of phase contributions from incomplete cancellation of eddy-currents or inaccurate pre-emphasis. Complex phase behaviour with increasing b-values was seen in the bipolar case while the unipolar

sequence lacked such crossings. This sequence difference is possibly related to the fact that there were more gradient switches in the diffusion-sensitizing gradients of the bipolar sequence, with eddy-currents arising from more time-points. The specific timing of gradient switches depended on the b-value. Eddy currents cancel each other if a gradient switch Nutlin-3a chemical structure is closely followed in time by an opposite gradient switch [17], [31], [32] and [33]. However, the

switching of strong gradients with relatively long temporal separation (as in diffusion imaging) results in incomplete cancellation and residual eddy currents. The linear accumulation of 0th-order phases could be related to a drift in the centre frequency between the calibration and phantom scans. The second- and third-order phases had relatively linear accrual that persisted beyond the readout. This suggests the presence of eddy currents with relatively long time constants. Compared to those with intermediate time constants, eddy-currents with longer time constants have better self-cancellation properties (following opposite gradient switches of trapezoidal diffusion pulses). However, neither will completely cancel out since the gradient switches are not coincident in time. The field AZD6244 camera is sensitive to small residual eddy-current phases resulting from incomplete cancellation Atezolizumab order [20], [34] and [35]. The gradient pre-emphasis

was on and its effects were included in the measured phases. Thus, any residual eddy currents contribute to the shape of the observed phases. More comprehensive models are required to fully describe eddy-current behaviour [34] and [35]. The gradient impulse response method is free from model restrictions and can measure residual eddy-currents phases that do not conform to those predicted by simple models with limited sets of exponential terms. In general, the specific shapes of the eddy-current phases can only be predicted closely by characterizing the entire frequency behaviour of the gradient system [34] and [35]. In a clinical setting, the TE would be determined by the maximum b-value in the set. The other (lower) b-values in the set would have lower gradient amplitudes and thus, less eddy current distortions. However, the purpose in this study was to measure the maximum eddy-current contribution (by applying the diffusion pulses at maximum gradient strength with shortest TE) to determine the worst case scenario at each chosen b-value.

The length of CKX ORF in foxtail millet ranged from 720 to 1620 bp. BLAST analysis against the Pfam and SMART database indicated that all of them belonged to the SiCKX gene family. The predicted SiCKX proteins had a typical FAD- and CK-binding domains, which were specific to CKX family members. The 11 SiCKX genes were distributed on seven foxtail millet chromosomes: chromosomes 1, 3, 4, 6, 7, and 11 each contains one gene, while chromosome 5 contains five genes. The tool of NetOGlyc (http://www.cbs.dtu.dk/services/NetOGlyc/) was used to predict Dabrafenib manufacturer the number of glycosylation sites. SiCKX1, SiCKX3, SiCKX5, and SiCKX10 each contains two glycosylation sites, SiCKX7, SiCKX8 and SiCKX11

each contains five glycosylation sites, SiCKX4 contains three sites, SiCKX6

contains one site and SiCKX2 and SiCKX9 contain no glycosylation sites. Five of the 11 SiCKX proteins showed localization on the chloroplast thylakoid membrane by a PSORT analysis (http://psort.nibb.ac.jp/). Of the remaining proteins, SiCKX2, SiCKX5 and SiCKX9 showed localization in the cytoplasm, SiCKX4 located in the nuclear, and SiCKX10 and SiCKX11 showed localization in the extracellular and vacuole, respectively ( Table 1). The cDNA sequences were compared with see more the corresponding genomic DNA sequences to detect the numbers and positions of exons and introns within each SiCKX gene by using the GSDS program (http://gsds.cbi.pku.edu.cn/) ( Fig. 1). The coding sequences of all the SiCKX genes were disrupted by introns, with numbers varying from one to four. The motif distribution in SiCKX proteins was analyzed based on the MEME program. Three putative conserved motifs were identified, each with 50 amino acids. All three were present in each SiCKX member except SiCKX9; motif 2 appeared twice in SiCKX8, and SiCKX9 contained only motif 1 and motif 2 ( Fig. 2). In order to uncover the evolutionary relationships among foxtail millet, rice and Arabidopsis

CKXs, the amino acid sequences of CKX genes were compared by ClustalX. In the phylogenetic tree constructed by the NJ method ( Fig. 3) the proteins clustered into three major groups (I, II, and III). Group I contained 22 members (with 8, 9, and 5 members of foxtail millet, rice, Arabidopsis, respectively) Idoxuridine and further divided into subclusters IA and IB. Group II included 6 members (with 3, 3, and 1 members of foxtail millet, rice, Arabidopsis, respectively). Group III contained the SiCKX7 gene only. Based on phylogenetic results (Fig. 4), four paralogs (SiCKX1/SiCKX3, SiCKX2/SiCKX4, SiCKX5/SiCKX8, and SiCKX10/SiCKX11), were identified in SiCKX genes. According to the foxtail millet genome annotation results, we found one tandemly duplicated pair, namely SiCKX5/SiCKX8, on chromosome 5. Segmental duplications might have contributed to the other three paralogous genes ( Fig. 5).

All cDNA was quantified using a NanoDrop Spectrophotometer – 2000 (NANODROP, USA). The concentrations were adjusted, and samples were stored at −80 °C. All gene expression was measured by qRT-PCR on the Applied Biosystems 7500 Fast Real-Time PCR system (Applied Biosystems™, USA), using the cycling conditions recommended by Applied Biosystems. We used the following assays: preproET-1 (ppET-1)– Assay ID: Rn00561129_m1*, ETA – Assay Id: Rn00561137_m1*, ETB – Assay Id: Rn00569139_m1* and GAPDH -

Assay ID: Rn99999916_s1. The threshold values were uniformly set for all assays. All reactions were performed in duplicate. Replicates with standard deviations (SD) higher than 0.5 for the cycle threshold (CT) value were repeated or excluded from the analysis. The amplification curve of each group was determined, and the CT values were obtained for all genes (ppET-1, ETA, ETB and GAPDH). We

used the comparative learn more CT method (ΔΔCT method), where we first calculated ΔCT = CT target – CT endogenous controls to normalize the target gene to the endogenous controls. Notably, the Relative Quantification (RQ) of ppET-1, ETA, ETB genes was calculated using the control group as a reference and using the 2-ΔΔCT formula, which provides the percentage change, or how much more one gene is expressed in one group relative to another. All CT values were obtained using 7500 software 2.0, and these data were exported to Microsoft Excel (Microsoft, USA) to calculate 2-ΔCT and RQ. The data are presented Montelukast Sodium as the mean ± SEM. The Rmax and pEC50 values were compared by see more two-way ANOVA followed by Bonferroni’s post-test because one variable was the physical training and the other was exposure to a single exercise session. P < 0.05 were considered statistically significant. The Ang II responses in femoral veins are discrete and difficult to measure. Therefore, the Ang II

concentration-response curves in the femoral veins are characteristically low. These curves exhibited a similar pattern in both sedentary and trained animals, whether studied at rest or after a single bout of exercise (Fig. 1A). Differences between groups were not observed in the presence of indomethacin either (Fig. 1B). In the presence of L-NAME, however, the Ang II concentration-response curves determined for resting-sedentary animals as well as the related Rmax values were higher compared to the other groups ( Fig. 1C). However, in the presence of both L-NAME and indomethacin, preparations taken from exercised-sedentary, resting-trained and exercised-trained animals exhibited Ang II concentration-response curves of similar magnitude to preparations taken from resting-sedentary animals ( Fig. 1D). Indeed, the difference in the Ang II Rmax observed between groups in the presence of L-NAME disappeared in the presence of both L-NAME and indomethacin.

For inoculating the fungus expressing DsRed, the maize roots 1 cm in length were immersed in a suspension of spores (105 conidia mL− 1) for 5 min before transfer

to 1 mL of BNM medium on a slide. The growth and colonization of F. verticillioides were subjected to epifluorescent microscopy. Following infection with F. verticillioides, the dyes of neutral red (0.01%, W/V) and Evans blue (0.2%, W/V) (Sigma, St. Louis, MO, USA) were used to stain the cross and longitudinal sections of the roots for 5 min each, rinsed with water, and then observed under a microscope. NVP-BEZ235 ic50 Dead cells were stained blue with Evans blue, whereas living cells were stained red with neutral red. Certain characteristic indicators, e.g., accumulation of peroxide, can be detected when PCD occurs. To investigate whether infection of F. verticillioides induced PCD in maize leaves, peroxide

staining using 3,3-diaminobenzidine (DAB) as the substrate was performed to detect the accumulation of H2O2 following infection of F. verticillioides as described previously [33]. At the two-leaf-stage, the leaves of maize plants inoculated with the F. verticillioides strain expressing DsRed were excised and incubated in a 1 mg mL− 1 solution of DAB (pH 3.8) for 2 h under light at 25 °C and then boiled in ethanol (96%) for 10 min. After cooling, the leaves were extracted with fresh ethanol at room temperature. The degree of dark brown polymerization indicated the amount of H2O2 accumulated in the treated leaves. Selleck Cabozantinib Selective Fusarium Methocarbamol agar (SFA) [29] and [30] medium was used to analyze the colonization of F. verticillioides on/in maize roots. DsRed-labeled fungus-infected and mock-inoculated roots and basal stems of maize were sampled at different times after the inoculation from two replicated greenhouse trials to determine the numbers of colony forming units (CFU) as previously described [31]. A randomized complete block design with four replicates consisting of two plants each was used to arrange the inoculated maize plants. The roots

were removed from the vermiculite, washed thoroughly with tap water, and surface sterilized for 3 min in 0.5% (V/V) NaOCl solution. After rinsing with sterile deionized water several times, roots were wiped with sterile filter paper. Roots and basal stems from the two plants in each replicate were weighted, ground, and mixed into 10 ml of sterile deionized water with a Fast-Prep-24 Instrument (MP Biomedicals, Solon, OH, USA) at high speed for 1 min. Homogenized suspensions of root and basal stem samples were filtered through four layers of cheesecloth to remove plant debris and diluted 20-fold with sterile deionized water. The diluted samples were separately spread with a sterile glass rod onto the SFA plates. Each inoculated sample consisted of five replicated plates with 50 μL of diluted tissue suspension.