Name: oscillation and reaction board techniques for estimating inertial properties of a below-knee prosthesis
Uploaded: 2014-05-08T16:00:00
Duration: 8 min 8 s
Description: Watch this Scientific Journal Video about Oscillation and Reaction Board Techniques for Estimating Inertial Properties of a Below-knee Prosthesis at JoVE.com

Funding from the American and International Societies of Biomechanics was provided for this study.

<ol>
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H., Kerwin, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+the+uncertainties+in+resultant+joint+moments+computed+in+a+dynamic+activity.">Quantification of the uncertainties in resultant joint moments computed in a dynamic activity.</a> J Sports Sci. 14, 219-231 (1996).</li><li>Hunter, J. P., Marshall, R. N., McNair, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Segment-interaction+analysis+of+the+stance+limb+in+sprint+running.">Segment-interaction analysis of the stance limb in sprint running.</a> J Biomech. 37, 1439-1446 (2004).</li><li>Lin-Chan, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+added+prosthetic+mass+on+physiologic+responses+and+stride+frequency+during+multiple+speeds+of+walking+in+persons+with+transtibial+amputation.">The effects of added prosthetic mass on physiologic responses and stride frequency during multiple speeds of walking in persons with transtibial amputation.</a> Arch Phys Med Rehabil. 84, 1865-1871 (2003).</li><li>Mattes, S. J., Martin, P. E., Royer, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Walking+symmetry+and+energy+cost+in+persons+with+unilateral+transtibial+amputations:+matching+prosthetic+and+intact+limb+inertial+properties.">Walking symmetry and energy cost in persons with unilateral transtibial amputations: matching prosthetic and intact limb inertial properties.</a> Arch Phys Med Rehabil. 81, 561-568 (2000).</li><li>Smith, J. D., Martin, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short+and+longer+term+changes+in+amputee+walking+patterns+due+to+increased+prosthesis+inertia.">Short and longer term changes in amputee walking patterns due to increased prosthesis inertia.</a> J Prosthet Orthot. 23, 114-123 (2011).</li><li>Smith, J. D., Martin, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+prosthetic+mass+distribution+on+metabolic+costs+and+walking+symmetry.">Effects of prosthetic mass distribution on metabolic costs and walking symmetry.</a> J Appl Biomech. 29, 317-328 (2013).</li><li>Czerniecki, J. M., Gitter, A., Munro, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Joint+moment+and+muscle+power+output+characteristics+of+below+knee+amputees+during+running:+the+influence+of+energy+storing+prosthetic+feet.">Joint moment and muscle power output characteristics of below knee amputees during running: the influence of energy storing prosthetic feet.</a> J Biomech. 24, 63-75 (1991).</li><li>Miller, D. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resultant+lower+extremity+joint+moments+in+below-knee+amputees+during+running+stance.">Resultant lower extremity joint moments in below-knee amputees during running stance.</a> J Biomech. 20, 529-541 (1987).</li><li>Vanicek, N., Strike, S., McNaughton, L., Polman, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gait+patterns+in+transtibial+amputee+fallers+vs.+non-fallers:+Biomechanical+differences+during+level+walking.">Gait patterns in transtibial amputee fallers vs. non-fallers: Biomechanical differences during level walking.</a> Gait &amp; Posture. 29, 415-420 (2009).</li><li>Royer, T., Koenig, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Joint+loading+and+bone+mineral+density+in+persons+with+unilateral,+trans-tibial+amputation.">Joint loading and bone mineral density in persons with unilateral, trans-tibial amputation.</a> Clin Biomech. 20, 1119-1125 (2005).</li><li>Underwood, H. 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The authors declare that they have no competing financial interests.

To quantify the resultant joint forces and moments during movement, an inverse dynamics model of the system of interest is needed when working with empirical data. For lower extremity biomechanics, inverse dynamics models typically represent the foot, shank, and thigh as rigid bodies. Input for these models come from three primary sources: a) motion kinematics, b) ground reaction forces, and c) segment anthropometrics and inertial properties. Motion data are collected with a variety of motion analysis systems, but all systems essentially provide the basic kinematics of the movement (position, velocity, and acceleration). Ground reaction forces are collected with a force plate and provide the contact forces acting on the feet. Anthropometrics are measurements taken directly from the body using rulers, flexible tapes, and/or calipers.&#160;These anthropometric measurements are used to estimate the inertial properties of the body segments used in the inverse dynamics analyses. Inertial properties include the mass, center of mass (COM) location and moment of inertia (MOI) of the segment relative to an axis through the segment COM or the proximal or distal joint. Methodologies and equipment used for collecting motion and ground reaction force data are similar among research groups, but inertial estimations of body segments can vary widely among researchers depending on which method the researcher chooses for estimating these inertial properties.
Various techniques available for estimating the inertial properties of a fully intact human body segment include: 1) regression equations based on cadaver data1-5, 2) mathematical models (i.e., geometric models)6,7, and 3) scanning &#38; imaging techniques8-15. Many of these techniques require direct measurements from the body, but it has previously been shown that regardless of the estimation method being used, the precision of body segment inertial estimates based on these methods is high16. It has also been shown that errors in the estimates of the inertial properties of intact body segments have minimal impact on the magnitudes of resultant joint moments during walking17,18. Joint moments are influenced to a greater extent by ground reaction forces, center of pressure locations, moment arm lengths, and segment kinematics17-19. Therefore, it is not surprising that methods for estimating inertial properties of body segments vary widely across the literature when using able-bodied individuals as research participants given that small errors in these estimates are likely to have little impact on outcomes of the study.
Many of these inertial estimations for a fully intact body segment are often used for estimating the inertial properties of prostheses for lower extremity amputees. Modern lower limb prostheses are fabricated using lightweight materials resulting in prosthetic limbs that are much lighter than the limbs they replace. This results in an inertial asymmetry between the prosthetic limb and intact limb. Compared with a typical intact shank and foot, the mass of a below-knee prosthesis and residual limb is approximately 35% less and has a center of mass located approximately 35% closer to the knee joint20-23. The lower mass and more proximal mass distribution of the prosthetic limb also produces a much lower (~60%) moment of inertia relative to the knee joint for the prosthetic limb compared to that of the intact shank and foot. Although researchers24,25 have previously suggested that using intact inertial estimates for the prosthetic limb have little effect on joint kinetic estimates, these comparisons focused on resultant joint moments during the stance phase of walking, where the ground reaction force dominates the moment produced at the joint. During swing, where ground reaction forces are not present, the reduced inertial properties of the prosthesis are more likely to influence estimates of resultant joint moments. Given that some researchers e.g., 26-32 utilize intact segment inertia properties to represent prosthesis inertial properties and otherse.g. 21-23 estimate prosthesis inertial properties directly, it is important to understand the impact of the methods chosen for estimating the inertial properties of the prosthesis. Minimizing the time required for measuring inertial properties of the prosthesis was an important consideration in the development of our technique. In the technique presented here the prosthesis remains fully intact for all measurements to reduce measurement times and avoid any additional times associated with realigning the prosthesis after measurement.
Thus, the purpose of this study was two-fold: 1) demonstrate a technique that can be used directly to estimate the inertial properties of a below-knee prosthesis, and 2) contrast the effects of the proposed technique and that of using intact limb inertial properties on joint kinetic estimates during walking in unilateral, transtibial amputees. It was hypothesized that joint kinetic magnitudes are larger when inertial properties of the intact shank and foot are used as the inertial estimates for the prosthesis compared to direct measurements of the prosthesis inertial properties.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr height="105">
			<td height="105" style="height:105px;width:216px;">Oscillation Rack &#38; Reaction Board</td>
			<td style="width:144px;">Custom Built</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Outer cage made from 80/20 aluminum, inner cage from various thicknesses of solid of aluminum.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Laboratory scale</td>
			<td style="width:144px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">NI LabView</td>
			<td style="width:144px;">National Instruments</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Software for recording TTL pulses from infrared photocell.</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">BNC-1050</td>
			<td style="width:144px;">National Instruments</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">BNC Breakout box with direct pin connections to the data acquisition card</td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:216px;">MATLAB</td>
			<td style="width:144px;">Mathworks Inc.</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Software for processing oscillation and reaction board data to predict inertial properties of prosthesis.</td>
		</tr>
	</tbody>
</table>

oscillation and reaction board techniques for estimating inertial properties of a below-knee prosthesis

The purpose of this study was two-fold: 1) demonstrate a technique that can be used to directly estimate the inertial properties of a below-knee prosthesis, and 2) contrast the effects of the proposed technique and that of using intact limb inertial properties on joint kinetic estimates during walking in unilateral, transtibial amputees. An oscillation and reaction board system was validated and shown to be reliable when measuring inertial properties of known geometrical solids. When direct measurements of inertial properties of the prosthesis were used in inverse dynamics modeling of the lower extremity compared with inertial estimates based on an intact shank and foot, joint kinetics at the hip and knee were significantly lower during the swing phase of walking. Differences in joint kinetics during stance, however, were smaller than those observed during swing. Therefore, researchers focusing on the swing phase of walking should consider the impact of prosthesis inertia property estimates on study outcomes. For stance, either one of the two inertial models investigated in our study would likely lead to similar outcomes with an inverse dynamics assessment.

Inertial properties of the prosthetic leg distal to the knee were lower than those of the intact leg (Table 1). Averaged across participants, prosthetic side mass was 39% less, moment of inertia about a transverse axis through the knee was 52% less, and the center of mass location was 24% closer to the knee compared with values for the intact leg.
<table border="1" cellpadding="0" cellspacing="0" fo:keep-together.within-page="always" width="553">
	<tbody>
		<tr>
			<td>Subject</td>
			<td>Intact* Mass (kg)</td>
			<td>Pros&#8224; Mass (kg)</td>
			<td>Est. Mass difference (kg)</td>
			<td>&#8225;Ikneeintact (kg&#183;m2)</td>
			<td>Ikneepros (kg&#183;m2)</td>
			<td>Intact CM below the knee joint (m)</td>
			<td>Pros CM below the knee joint (m)</td>
		</tr>
		<tr>
			<td>A</td>
			<td>6.03</td>
			<td>4.27</td>
			<td>1.76</td>
			<td>0.604</td>
			<td>0.325</td>
			<td>0.268</td>
			<td>0.215</td>
		</tr>
		<tr>
			<td>B</td>
			<td>6.07</td>
			<td>3.39</td>
			<td>2.68</td>
			<td>0.400</td>
			<td>0.196</td>
			<td>0.215</td>
			<td>0.177</td>
		</tr>
		<tr>
			<td>C</td>
			<td>5.80</td>
			<td>3.12</td>
			<td>2.68</td>
			<td>0.575</td>
			<td>0.194</td>
			<td>0.264</td>
			<td>0.198</td>
		</tr>
		<tr>
			<td>D</td>
			<td>5.72</td>
			<td>3.17</td>
			<td>2.55</td>
			<td>0.559</td>
			<td>0.317</td>
			<td>0.265</td>
			<td>0.191</td>
		</tr>
		<tr>
			<td>E</td>
			<td>7.14</td>
			<td>4.65</td>
			<td>2.49</td>
			<td>0.742</td>
			<td>0.325</td>
			<td>0.276</td>
			<td>0.200</td>
		</tr>
		<tr>
			<td>F</td>
			<td>6.23</td>
			<td>4.22</td>
			<td>2.01</td>
			<td>0.585</td>
			<td>0.287</td>
			<td>0.260</td>
			<td>0.192</td>
		</tr>
		<tr>
			<td>Mean &#177; STD</td>
			<td>6.17 &#177; 0.51</td>
			<td>3.80 &#177; 0.66</td>
			<td>2.36 &#177; 0.38</td>
			<td>0.578 &#177; 0.109</td>
			<td>0.274 &#177; 0.063</td>
			<td>0.258 &#177; 0.022</td>
			<td>0.196 &#177; 0.013</td>
		</tr>
	</tbody>
</table>
*Intact refers to values for the combined intact shank and foot. 
&#8224;Pros refers to values for the combined prosthesis and residual limb. 
&#8225;Moment of inertia about a transverse axis through the knee.
Table 1. Comparison of inertial properties between the prosthetic and intact limbs from the knee down.
Resultant joint forces (Figure 3) and moments (Figure 4) at the ankle, knee, and hip were affected by the inertial parameters used in the inverse dynamics model. Specifically, joint kinetics were reduced during swing initiation (~65% of the gait cycle) and swing termination (~95% of the gait cycle) when direct measures of prosthesis inertia were used in inverse dynamics assessments compared with regressions based on intact anatomy (Table 2). Effect sizes suggest these differences during swing were not trivial (&#8805;1.0). In addition, joint kinetic values increased on average by 80% during swing initiation and termination when intact inertial estimates were used compared with direct measures of the prosthesis inertial properties. Thus, when inertial properties of an intact limb were used to model the prosthetic side, joint kinetic profiles of the prosthetic side during swing more closely resembled those of an intact limb (see Figures 3 and 4). During stance, a number of statistical differences were observed. The largest effect size for any difference during stance was observed for the hip anterioposterior resultant joint force (ES = 0.86). Although this effect size is large and still considered as part of stance, the peak value for this measure occurred during terminal stance (~52%), or as the limb was transitioning into swing. Effect sizes for all other significant differences observed during stance ranged from 0.01 to 0.41, which would be considered small effects with the larger of these values being observed in the hip resultant joint reaction forces. Although significant differences were found during stance, these differences when considered in terms of the size of the difference (i.e., effect sizes) might lead one to question the meaningfulness of these differences.
<img alt="Figure 3" fo:content-width="5in" src="/files/ftp_upload/50977/50977fig3highres.jpg" width="500" /> 
Figure 3. Resultant joint reaction forces of the ankle, knee, and hip in the anterioposterior (left panels) and vertical directions (right panels). Data were averaged across subjects for presentation. The stance phase begins at 0% of the gait cycle with foot contact and ends at approximately 60% of the gait cycle with toe-off. Swing continues until the next foot contact of the same leg at 100% of the gait cycle. <a href="https://www.jove.com/files/ftp_upload/50977/50977fig3highres.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 4" fo:content-width="5in" src="/files/ftp_upload/50977/50977fig4highres.jpg" width="500" /> 
Figure 4. Resultant joint moments about a transverse axis (a.k.a, mediolateral axis) through the ankle, knee, and hip. Data were averaged across subjects for presentation. The stance phase begins at 0% of the gait cycle with foot contact and ends at approximately 60% of the gait cycle with toe-off. Swing continues until the next foot contact of the same leg at 100% of the gait cycle.
<img alt="Table 2" fo:content-width="6in" src="/files/ftp_upload/50977/50977table2.jpg" width="600" /> 
Table 2. Peak resultant joint reaction forces and moments averaged across subjects and statistical comparisons between the two inertial models for the prosthetic side's joint kinetics. Notes: Mean data are presented as mean(SD). % Gait Cycle column represents the average percentage across subjects where the peak value occurred for that variable. p&#60;.05 considered significant.

Watch this Scientific Journal Video about Oscillation and Reaction Board Techniques for Estimating Inertial Properties of a Below-knee Prosthesis at JoVE.com

Oscillation and Reaction Board Techniques for Estimating Inertial Properties of a Below-knee Prosthesis

Body segmental inertial properties are required for inverse dynamics modeling. Using an oscillation and reaction board technique, inertial properties of below-knee prostheses were measured. Using direct measures of prosthesis inertia in the inverse dynamics model of the prosthetic leg resulted in lower magnitudes of resultant joint forces and moments.

The purpose of this study was two-fold: 1) demonstrate a technique that can be used to directly estimate the inertial properties of a below-knee ...

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Measuring the Mechanical Properties of Living Cells Using Atomic Force Microscopy

Mechanical properties of cells and extracellular matrix (ECM) play important roles in many biological processes including stem cell differentiation, tumor formation, and wound healing. Changes in stiffness of cells and ECM are often signs of changes in cell physiology or diseases in tissues. Hence, cell stiffness is an index to evaluate the status of cell cultures. Among the multitude of methods applied to measure the stiffness of cells and tissues, micro-indentation using an Atomic Force Microscope (AFM) provides a way to reliably measure the stiffness of living cells. This method has been widely applied to characterize the micro-scale stiffness for a variety of materials ranging from metal surfaces to soft biological tissues and cells. The basic principle of this method is to indent a cell with an AFM tip of selected geometry and measure the applied force from the bending of the AFM cantilever. Fitting the force-indentation curve to the Hertz model for the corresponding tip geometry can give quantitative measurements of material stiffness. This paper demonstrates the procedure to characterize the stiffness of living cells using AFM. Key steps including the process of AFM calibration, force-curve acquisition, and data analysis using a MATLAB routine are demonstrated. Limitations of this method are also discussed.

This paper demonstrates a protocol to characterize the mechanical properties of living cells by means of microindentation using an Atomic Force Microscope (AFM).

Mechanical  properties of cells and extracellular matrix (ECM) play important roles in many  biological processes including stem cell differentiation, ...

Engineering Platform and Experimental Protocol for Design and Evaluation of a Neurally-controlled Powered Transfemoral Prosthesis

To enable intuitive operation of powered artificial legs, an interface between user and prosthesis that can recognize the user's movement intent is desired. A novel neural-machine interface (NMI) based on neuromuscular-mechanical fusion developed in our previous study has demonstrated a great potential to accurately identify the intended movement of transfemoral amputees. However, this interface has not yet been integrated with a powered prosthetic leg for true neural control. This study aimed to report (1) a flexible platform to implement and optimize neural control of powered lower limb prosthesis and (2) an experimental setup and protocol to evaluate neural prosthesis control on patients with lower limb amputations. First a platform based on a PC and a visual programming environment were developed to implement the prosthesis control algorithms, including NMI training algorithm, NMI online testing algorithm, and intrinsic control algorithm. To demonstrate the function of this platform, in this study the NMI based on neuromuscular-mechanical fusion was hierarchically integrated with intrinsic control of a prototypical transfemoral prosthesis. One patient with a unilateral transfemoral amputation was recruited to evaluate our implemented neural controller when performing activities, such as standing, level-ground walking, ramp ascent, and ramp descent continuously in the laboratory. A novel experimental setup and protocol were developed in order to test the new prosthesis control safely and efficiently. The presented proof-of-concept platform and experimental setup and protocol could aid the future development and application of neurally-controlled powered artificial legs.

Neural-machine interfaces (NMI) have been developed to identify the user's locomotion mode. These NMIs are potentially useful for neural control of powered artificial legs, but have not been fully demonstrated. This paper presented (1) our designed engineering platform for easy implementation and development of neural control for powered lower limb prostheses and (2) an experimental setup and protocol in a laboratory environment to evaluate neurally-controlled artificial legs on patients with lower limb amputations safely and efficiently.

To enable intuitive operation of powered artificial legs, an interface between user and prosthesis that can recognize the user's movement intent is ...

In Situ Mapping of the Mechanical Properties of Biofilms by Particle-tracking Microrheology

Bacterial cells are able to form surface-attached biofilm communities known as biofilms by encasing themselves in extracellular polymeric substances (EPS). The EPS serves as a physical and protective scaffold that houses the bacterial cells and consists of a variety of materials that includes proteins, exopolysaccharides and DNA. The composition of the EPS may change, which remodels the mechanic properties of the biofilm to further develop or support alternative biofilm structures, such as streamers, as a response to environmental cues. Despite this, there are little quantitative descriptions on how EPS components contribute to the mechanical properties and function of biofilms. Rheology, the study of the flow of matter, is of particular relevance to biofilms as many biofilms grow in flow conditions and are constantly exposed to shear stress. It also provides measurement and insight on the spreading of the biofilm on a surface. Here, particle-tracking microrheology is used to examine the viscoelasticity and effective crosslinking roles of different matrix components in various parts of the biofilm during development. This approach allows researchers to measure mechanic properties of biofilms at the micro-scale, which might provide useful information for controlling and engineering biofilms.

In Situ Mapping of the Mechanical Properties of Biofilms by Particle-tracking Microrheology

Particle-tracking microrheology investigates the viscoelasticity of materials. Here, the technique is used to determine the viscoelasticity, creep compliance and effective crosslinking roles of different matrix components of a bacterial biofilm. The matrix consists of polymeric substances secreted by the bacteria and its components determine biofilm structure and mechanical properties.

Bacterial cells are able to form surface-attached biofilm communities known as biofilms by encasing themselves in extracellular polymeric substances ...

Techniques for the Evolution of Robust Pentose-fermenting Yeast for Bioconversion of Lignocellulose to Ethanol

Lignocellulosic biomass is an abundant, renewable feedstock useful for production of fuel-grade ethanol and other bio-products. Pretreatment and enzyme saccharification processes release sugars that can be fermented by yeast. Traditional industrial yeasts do not ferment xylose (comprising up to 40% of plant sugars) and are not able to function in concentrated hydrolyzates. Concentrated hydrolyzates are needed to support economical ethanol recovery, but they are laden with toxic byproducts generated during pretreatment. While detoxification methods can render hydrolyzates fermentable, they are costly and generate waste disposal liabilities. Here, adaptive evolution and isolation techniques are described and demonstrated to yield derivatives of the native Scheffersomyces stipitis strain NRRL Y-7124 that are able to efficiently convert hydrolyzates to economically recoverable ethanol despite adverse culture conditions. Improved individuals are enriched in an evolving population using multiple selection pressures reliant on natural genetic diversity of the S. stipitis population and mutations induced by exposures to two diverse hydrolyzates, ethanol or UV radiation. Final evolution cultures are dilution plated to harvest predominant isolates, while intermediate populations, frozen in glycerol at various stages of evolution, are enriched on selective media using appropriate stress gradients to recover most promising isolates through dilution plating. Isolates are screened on various hydrolyzate types and ranked using a novel procedure involving dimensionless relative performance index (RPI) transformations of the xylose uptake rate and ethanol yield data. Using the RPI statistical parameter, an overall relative performance average is calculated to rank isolates based on multiple factors, including culture conditions (varying in nutrients and inhibitors) and kinetic characteristics. Through application of these techniques, derivatives of the parent strain had the following improved features in enzyme saccharified hydrolyzates at pH 5-6: reduced initial lag phase preceding growth, reduced diauxic lag during glucose-xylose transition, significantly enhanced fermentation rates, improved ethanol tolerance and accumulation to 40 g/L.

Adaptive evolution and isolation techniques are described and demonstrated to yield derivatives of Scheffersomyces stipitis strain NRRL Y-7124 that are able to rapidly consume hexose and pentose mixed sugars in enzyme saccharified undetoxified hydrolyzates and to accumulate over 40 g/L ethanol.

Lignocellulosic biomass is an abundant, renewable feedstock useful for production of fuel-grade ethanol and other bio-products. Pretreatment and enzyme ...

Production of Metal Nanoparticles by Pulsed Laser-ablation in Liquids: A Tool for Studying the Antibacterial Properties of Nanoparticles

The emergence of multidrug-resistant bacteria is a global clinical concern leading some to speculate about our return to a "pre-antibiotics" era of medicine. In addition to efforts to identify novel small-molecule antimicrobial drugs, there has been great interest in the use of metal nanoparticles as coatings for medical devices, wound dressings, and consumer packaging, due to their antimicrobial properties. The wide variety of methods available for nanoparticle synthesis results in a broad spectrum of chemical and physical properties which can affect antibacterial efficacy. This manuscript describes the pulsed laser-ablation in liquids (PLAL) method to create nanoparticles. This approach allows for the fine tuning of nanoparticle size, composition, and stability using post-irradiation methods as well as the addition of surfactants or volume excluders. By controlling particle size and composition, a large range of physical and chemical properties of metal nanoparticles can be explored which may contribute to their antimicrobial efficacy thereby opening new avenues for antibacterial development.

The antimicrobial properties of metals such as copper and silver have been recognized for centuries. This protocol describes pulsed laser-ablation in liquids, a method of synthesizing metal nanoparticles that provides the ability to fine tune the properties of these nanoparticles to optimize their antimicrobial effects.

The emergence of multidrug-resistant bacteria is a global clinical concern leading some to speculate about our return to a "pre-antibiotics" era of ...

Synthesis of Hydrogels with Antifouling Properties As Membranes for Water Purification

Hydrogels have been widely utilized to enhance the surface hydrophilicity of membranes for water purification, increasing the antifouling properties and thus achieving stable water permeability through membranes over time. Here, we report a facile method to prepare hydrogels based on zwitterions for membrane applications. Freestanding films can be prepared from sulfobetaine methacrylate (SBMA) with a crosslinker of poly(ethylene glycol) diacrylate (PEGDA) via photopolymerization. The hydrogels can also be prepared by impregnation into hydrophobic porous supports to enhance the mechanical strength. These films can be characterized by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) to determine the degree of conversion of the (meth)acrylate groups, using goniometers for hydrophilicity and differential scanning calorimetry (DSC) for polymer chain dynamics. We also report protocols to determine the water permeability in dead-end filtration systems and the effect of foulants (bovine serum albumin, BSA) on membrane performance.

This paper reports practical methods to prepare hydrogels in freestanding films and impregnated membranes and to characterize their physical properties, including water transport properties.

Hydrogels have been widely utilized to enhance the surface hydrophilicity of membranes for water purification, increasing the antifouling properties and ...

Semiautomated Longitudinal Microcomputed Tomography-based Quantitative Structural Analysis of a Nude Rat Osteoporosis-related Vertebral Fracture Model

Osteoporosis-related vertebral compression fractures (OVCFs) are a common and clinically unmet need with increasing prevalence as the world population ages. Animal OVCF models are essential to the preclinical development of translational tissue engineering strategies. While a number of models currently exist, this protocol describes an optimized method for inducing multiple highly reproducible vertebral defects in a single nude rat. A novel longitudinal semiautomated microcomputed tomography (&#181;CT)-based quantitative structural analysis of the vertebral defects is also detailed. Briefly, rats were imaged at multiple time points post-op. The day 1 scan was reoriented to a standard position, and a standard volume of interest was defined. Subsequent &#181;CT scans of each rat were automatically registered to the day 1 scan so the same volume of interest was then analyzed to assess for new bone formation. This versatile approach can be adapted to a variety of other models where longitudinal imaging-based analysis could benefit from precise 3D semiautomated alignment. Taken together, this protocol describes a readily quantifiable and easily reproducible system for osteoporosis and bone research. The suggested protocol takes 4 months to induce osteoporosis in nude ovariectomized rats and between 2.7 and 4 h to generate, image, and analyze two vertebral defects, depending on tissue size and equipment.

The goal of this protocol is to generate a nude rat osteoporosis-related vertebral compression fracture model that can be longitudinally evaluated in vivo using a semiautomated microcomputed tomography-based quantitative structural analysis.

Osteoporosis-related vertebral compression fractures (OVCFs) are a common and clinically unmet need with increasing prevalence as the world population ...

A Millimeter Scale Flexural Testing System for Measuring the Mechanical Properties of Marine Sponge Spicules

Many load bearing biological structures (LBBSs)&#8212;such as feather rachises and spicules&#8212;are small (&#60;1 mm) but not microscopic. Measuring the flexural behavior of these LBBSs is important for understanding the origins of their remarkable mechanical functions.
We describe a protocol for performing three-point bending tests using a custom-built mechanical testing device that can measure forces ranging from 10-5 to 101 N and displacements ranging from 10-7 to 10-2 m. The primary advantage of this mechanical testing device is that the force and displacement capacities can be easily adjusted for different LBBSs. The device&#39;s operating principle is similar to that of an atomic force microscope. Namely, force is applied to the LBBS by a load point that is attached to the end of a cantilever. The load point displacement is measured by a fiber optic displacement sensor and converted into a force using the measured cantilever stiffness. The device&#39;s force range can be adjusted by using cantilevers of different stiffnesses.
The device&#39;s capabilities are demonstrated by performing three-point bending tests on the skeletal elements of the marine sponge Euplectella aspergillum. The skeletal elements&#8212;known as spicules&#8212;are silica fibers that are approximately 50 &#181;m in diameter. We describe the procedures for calibrating the mechanical testing device, mounting the spicules on a three-point bending fixture with a &#8776;1.3 mm span, and performing a bending test. The force applied to the spicule and its deflection at the location of the applied force are measured.

We present a protocol for performing three-point bending tests on sub-millimeter scale fibers using a custom-built mechanical testing device. The device can measure forces ranging from 20 &#181;N up to 10 N and can therefore accommodate a variety of fiber sizes.

Many load bearing biological structures (LBBSs)&#8212;such as feather rachises and spicules&#8212;are small (&#60;1 mm) but not microscopic. Measuring the ...

Synthesis of Multi-walled Carbon Nanotubes Modified with Silver Nanoparticles and Evaluation of Their Antibacterial Activities and Cytotoxic Properties

In this study, multi-walled carbon nanotubes (MWCNTs) were treated with an aqueous sulfuric acid solution to form an oxygen-based functional group. Silver MWCNTs were prepared by the reductive deposition of silver from an aqueous solution of AgNO3 on the oxidized MWCNTs. Given the unique color of the CNTs, it was not possible to apply them to the minimum inhibitory concentration or mitochondrial toxicity assays to evaluate the toxicity and antibacterial properties, since they would interfere with the assays. The inhibition zone and minimum bactericidal concentration for the Ag-MWCNTs were measured and Live/Dead and Trypan Blue assays were used to measure the toxicity and antibacterial properties without interfering with the color of the CNTs.

In this study, antimicrobial nanomaterials were synthesized by acidic oxidation of multiwalled carbon nanotubes and subsequent reductive deposition of silver nanoparticles. Antimicrobial activity and cytotoxicity tests were performed with the as-prepared nanomaterials.

In this study, multi-walled carbon nanotubes (MWCNTs) were treated with an aqueous sulfuric acid solution to form an oxygen-based functional group. Silver ...

3D Printing of In Vitro Hydrogel Microcarriers by Alternating Viscous-Inertial Force Jetting

Microcarriers are beads with a diameter of 60-250 &#181;m and a large specific surface area, which are commonly used as carriers for large-scale cell cultures. Microcarrier culture technology has become one of the main techniques in cytological research and is commonly used in the field of large-scale cell expansion. Microcarriers have also been shown to play an increasingly important role in in vitro tissue engineering construction and clinical drug screening. Current methods for preparing microcarriers include microfluidic chips and inkjet printing, which often rely on complex flow channel design, an incompatible two-phase interface, and a fixed nozzle shape. These methods face the challenges of complex nozzle processing, inconvenient nozzle changes, and excessive extrusion forces when applied to multiple bioink. In this study, a 3D printing technique, called alternating viscous-inertial force jetting, was applied to enable the construction of hydrogel microcarriers with a diameter of 100-300 &#181;m. Cells were subsequently seeded on microcarriers to form tissue engineering modules. Compared to existing methods, this method offers a free nozzle tip diameter, flexible nozzle switching, free control of printing parameters, and mild printing conditions for a wide range of bioactive materials.

3D Printing of In Vitro Hydrogel Microcarriers by Alternating Viscous-Inertial Force Jetting

Presented here is a mild 3D printing technique driven by alternating viscous-inertial forces to enable the construction of hydrogel microcarriers. Homemade nozzles offer flexibility, allowing easy replacement for different materials and diameters. Cell binding microcarriers with a diameter of 50-500 &#181;m can be obtained and collected for further culturing.

Microcarriers are beads with a diameter of 60-250 &#181;m and a large specific surface area, which are commonly used as carriers for large-scale cell ...