Simultaneous in vivo recording of prompt and delayed fluorescence and 820-nm reflection changes during drying and after rehydration of the resurrection plant Haberlea rhodopensis

A new instrument (M-PEA), which measures simultaneously kinetics of prompt fluorescence (PF), delayed fluorescence (DF) and modulated light reflection at 820 nm (MR), was used to screen dark-adapted leaves of the resurrection plant Haberlea rhodopensis during their progressive drying, down to 1% relative water content (RWC), and after their re-watering. This is the first investigation using M-PEA, which employs alternations of actinic light (627-nm peak, 5000 μmol photons m^? 2 s^? 1) and dark intervals, where PF-MR and DF kinetics are respectively recorded, with the added advantages: (a) all kinetics are recorded with high time resolution (starting from 0.01 ms), (b) the dark intervals' duration can be as short as 0.1 ms, (c) actinic illumination can be interrupted at different times during the PF transient (recorded up to 300 s), with the earliest interruption at 0.3 ms. Analysis of the simultaneous measurements at different water-content-states of H. rhodopensis leaves allowed the comparison and correlation of complementary information on the structure/function of the photosynthetic machinery, which is not destroyed but only inactivated (reversibly) at different degrees; the comparison and correlation helped also to test current interpretations of each signal and advance their understanding. Our results suggest that the desiccation tolerance of the photosynthetic machinery in H. rhodopensis is mainly based on mechanism(s) that lead to inactivation of photosystem II reaction centres (transformation to heat sinks), triggered already by a small RWC decrease.     

Nanosecond electric pulses induce DNA breaks in cisplatin-sensitive and -resistant human ovarian cancer cells

Human ovarian cancer cells COC1 and COC1/DDP (cisplatin-resistant subline) were exposed to 6 kV/cm nanosecond electric pulses (nsEP) with a pulse length of 8, 16 or 24 ns. The potential in a subcellular unit was calculated using a multilayer dielectric spherical model, and area under the voltage–time curves (AUC) integrated with a lower limit of 0.2 V. Cell viability was determined, and double-stand and total DNA breaks detected with the neutral and alkaline comet assays. nsEP evoked a higher voltage and AUC in nucleoplasm, and the levels in COC1 cells was just above those in COC1/DDP cells. Comets only appeared in the alkaline assay demonstrating single-stand DNA break. Fewer DNA break (16.51% vs. 35.13% at 24 ns, p = 0.0150) and more survival (22.42% vs. 13.19% at 24 ns, p = 0.0015) occurred in COC1/DDP cells despite an equal electric energy and almost equal cell sizes. 24-ns EP led to higher rates of cell-death and comet. The comet rate correlated with cell-death fraction in either cell line (r = 0.5701, p = 0.0135; r = 0.5110, p = 0.0302). There was no a correlation between the tail length, tail moment or Olive tail moment and cell-death rate. The data showed that response of chemosensitive cells differed from that of chemoresistant cells and DNA damage contributed to percent of cell death.     

Torque-onset determination: Unintended consequences of the threshold method

Background: Compared with visual torque-onset-detection (TOD), threshold-based TOD produces onset bias, which increases with lower torques or rates of torque development (RTD). Purpose: To compare the effects of differential TOD-bias on common contractile parameters in two torque-disparate groups. Methods: Fifteen boys and 12 men performed maximal, explosive, isometric knee-extensions. Torque and EMG were recorded for each contraction. Best contractions were selected by peak torque (MVC) and peak RTD. Visual-TOD-based torque-time traces, electromechanical delays (EMD), and times to peak RTD (tRTD) were compared with corresponding data derived from fixed 4-N m- and relative 5%MVC-thresholds. Results: The 5%MVC TOD-biases were similar for boys and men, but the corresponding 4-N m-based biases were markedly different (40.3 ± 14.1 vs. 18.4 ± 7.1 ms, respectively; p < 0.001). Boys–men EMD differences were most affected, increasing from 5.0 ms (visual) to 26.9 ms (4 N m; p < 0.01). Men’s visually-based torque kinetics tended to be faster than the boys’ (NS), but the 4-N m-based kinetics erroneously depicted the boys as being much faster to any given %MVC (p < 0.001). Conclusions: When comparing contractile properties of dissimilar groups, e.g., children vs. adults, threshold-based TOD methods can misrepresent reality and lead to erroneous conclusions. Relative-thresholds (e.g., 5% MVC) still introduce error, but group-comparisons are not confounded.     

Effects of stance width on postural movement pattern and anticipatory postural control associated with unilateral arm abduction

We investigated the effects of stance width on postural movement pattern and activation timing of postural muscles during unilateral arm abduction. Thirty-two healthy subjects abducted the right arm at their own timing. Stance width was 0, 9, 18 or 27 cm. Movement angles of leg lateral inclination and trunk lateral flexion to the leg in the frontal plane were analyzed. Based on movement angles at 0 cm width, subjects were classified into three groups: contralateral whole body leaning (CWBLg); ipsilateral trunk flexion (ITFg); and contralateral trunk flexion (CTFg). A high correlation between the movement angles was obtained at 0 cm width (r = 0.82). With increasing stance width, postural movement pattern in the ITFg shifted to patterns characterized by lateral flexion of the trunk toward the side opposite to arm movement, and movement angle of leg-inclination in ITFg and CWBLg decreased. At 0 cm width, left gluteus medius and tensor fascia latae were activated significantly about 40 ms ahead of the right middle deltoid in CWBLg and CTFg, but not in ITFg. However, preceding activation became prominent (about 20 ms) in ITFg for wide stances. Moreover, bilateral activation of the tensor fascia latae was observed in CTFg for all widths.     

Effect of image quality on correlation modeling error using a fiducial marker in a gimbaled linear accelerator

The purpose of this study was to investigate the effect of image quality under various imaging parameters (60, 70, 80, 90, 100, 110, and 120 kV at 200 mA and 10 ms/63, 80, 100, 160, 200, 250, and 320 mA at 120 kV and 10 ms) and the diameter of the fiducial marker (0.25, 0.50, 0.75, and 1.10 mm) on the correlation modeling error for dynamic tumor tracking (DTT) in the Vero4DRT system. Each fiducial marker was inserted into the center of the 30 × 30 × 10 cm³ water-equivalent phantom. A programmable respiratory motion table was used to simulate breathing-induced organ motion, with an amplitude of ±20 mm and a breathing cycle of 4 s. The correlation modeling error was calculated from the absolute difference between the detected and predicted target positions in the cranio-caudal direction. The image contrast of the fiducial marker was enhanced with increasing kV and mA. Increasing the diameter of the fiducial marker also enhanced the image contrast. Correlation-modeling error does not depend on the image quality and fiducial marker diameter. A lower kV setting did not generate a 4D model due to poor image contrast. All fiducial marker diameters were identified as good candidates for DTT in the Vero4DRT system.     

Effects of boldine on tyrosinase: Inhibition kinetics and computational simulation

Boldine is one of the most potent natural antioxidants and displays some important pharmacological activities, such as cytoprotective and anti-inflammatory activities. Based on its antioxidant properties, we studied the effects of boldine on l-DOPA oxidation by evaluating the inhibitory kinetics and a computational simulation between boldine and tyrosinase. Boldine reversibly inhibited tyrosinase from mushroom (Agaricus bisporus) in a mixed-type manner, with a K_i = 7.203 ± 0.933 mM. To gain insight into the inactivation process, we computed the kinetics via time-interval measurements and continuous substrate reactions. The results indicated that the inactivation induced by boldine was a first-order reaction with biphasic processes and that the substrate can promote the inactivation process. To gain further insight, we performed computational docking and molecular dynamics simulations, and the results showed that boldine can interact with several residues near the tyrosinase active site. Our study provides insight into the inhibition of tyrosinase in response to alkaloids. Based on its tyrosinase-inhibiting effect and low toxicity, boldine is a potential natural anti-pigmentation agent.     

Efficient transformation of mutant cells of Chlamydomonas reinhardtii by electroporation

Cells of CW-15 mutant of Chlamydomonas reinhardtii without a cell wall were transformed by electroporation. The hpt gene of hygromycin phosphotransferase was used as a selective marker. Optimal conditions of transformation were observed in the middle of the logarithmic growth phase at the density of suspension 10⁶ cells/ml, electric field intensity 1 kV/cm, and pulse duration 2 ms. Under these conditions up to 10³ hygromycin-resistant clones of trasformants per 10⁶ recipient cells were obtained that was 100 times higher than at the usage of wild-type cells. Exogenic DNA integrated into the genome of the nucleus C. reinhardtii was constantly inherited for more than 350 generations. The use of mutants without a cell wall and certain selective systems enable the efficiency of transformant yield to be doubled problems on unstable expression of geterologous genes to be investigated, and ways of obtaining super producers of foreign proteins using the alga C. reinhardtii investigated.     

Neural control of leg stiffness during hopping in boys and men

The purpose of the study was to investigate whether boys and men utilise different control strategies whilst hopping. Eleven boys (11–12 yr old) and ten men completed hopping at 1.5 Hz, 3.0 Hz and at their preferred frequency. A footswitch measured contact and flight times, from which leg stiffness was calculated. Simultaneously, surface electromyograms (EMGs) of selected lower limb muscles were recorded and quantified for each 30 ms period during the first 120 ms post-ground contact. At 1.5 Hz there were no differences between the groups in relative stiffness or muscle activity. At 3.0 Hz men had significantly shorter contact times (P = 0.013), longer flight times (P = 0.002), greater relative stiffness (P = 0.01) and significantly greater soleus (P = 0.012) and vastus lateralis (P < 0.001) activity during the initial 30 ms post-ground contact. At the preferred frequency men hopped significantly faster than the boys (P = 0.007), with greater leg stiffness (P < 0.01) and with more extensor activity in most time periods. Boys and men demonstrated similar control strategies when hopping at a slow frequency, but when hopping frequency increased men were able to better increase feedforward and reflex muscle activity to hop with greater relative stiffness.     

Descriptive analysis of kinematics and kinetics of catchers throwing to second base from their knees

In order to decrease the amount of time that it takes the catcher to throw the ball, a catcher may chose to throw from the knees. Upper extremity kinematics may play a significant role in the kinetics about the elbow observed in catchers throwing from the knees. If relationships between kinematics and kinetics exist then the development of training and coaching instruction may help in reduced upper extremity injury risk. Twenty-two baseball and softball catchers (14.36 ± 3.86 years; 165.11 ± 17.54 cm; 65.67 ± 20.60 kg) volunteered. The catchers exhibited a less trunk rotation (5.6 ± 16.2°), greater elbow flexion (87.9 ± 21.4°) and decreased humeral elevation (71.1 ± 12.3°) at the event of maximum shoulder external rotation as compared to what has previously reported in catchers. These variables are important, as they have previously been established as potential injury risk factors in pitchers, however it is not yet clear the role these variables play in catchers’ risk of injury. A positive relationship between elbow varus torque during the deceleration phase and elbow flexion at MIR was observed (r = 0.609; p = 0.003). Throwing from the knees reduces a catcher’s ability to utilize the proximal kinetic chain and this may help to explain why their kinematics and kinetics differ from what has previously been presented in the literature.     

Effect of ultrasound combined with malic acid on the activity and conformation of mushroom (Agaricus bisporus) polyphenoloxidase

Polyphenoloxidase (PPO) plays an important role in the browning of vegetables, fruits and edible fungi. The effects of ultrasound, malic acid, and their combination on the activity and conformation of mushroom (Agaricus bisporus) PPO were studied. The activity of PPO decreased gradually with the increasing of malic acid concentrations (5–60 mM). Neither medium concentrations (10, 20, 30 mM) malic acid nor individual ultrasound (25 kHz, 55.48 W/cm²) treatment could remarkably inactivate PPO. However, the inactivation during their combination was more significant than the sum of ultrasound inactivation and malic acid inactivation. The inactivation kinetics of PPO followed a first-order kinetics under the combination of ultrasound and malic acid. The conformation of combination treated PPO was changed, which was reflected in the decrease of α-helix, increase of β-sheet contents and disruption of the tertiary structure. Results of molecular microstructure showed that ultrasound broke large molecular groups of PPO into small ones. Moreover, combined treatment disrupted the microstructure of PPO and molecules were connected together.     

The inhibitory effect of curcumin on voltage-dependent K+ channels in rabbit coronary arterial smooth muscle cells

We investigated the effects of curcumin, the principal active compound of turmeric, on voltage-dependent K⁺ (Kv) channels in freshly isolated rabbit coronary arterial smooth muscle cells using the voltage-clamp technique. Curcumin reduced the Kv current in a dose-dependent manner with an apparent K_d value of 1.07 ± 0.03 μM. Although curcumin did not alter the kinetics of Kv current activation, it predominantly accelerated the decay rate of channel inactivation. The association and dissociation rate constants of curcumin were 1.35 ± 0.05 μM^?1 s^?1 and 1.47 ± 0.17 s^?1, respectively. Curcumin did not alter the steady-state activation or inactivation curves. Application of train pulses (1 or 2 Hz) increased curcumin-induced blockade of the Kv current, and the recovery time constant also increased in the presence of curcumin suggesting, that the inhibitory action of Kv currents by curcumin was use-dependent. From these results, we concluded that curcumin inhibited vascular Kv current in a state-, time-, and use-dependent manner.     

Effects of gender and age on median and ulnar nerve sensory responses over ring finger

ObjectiveTo analyze the effect of gender on median nerve (MN) and ulnar nerve (UN) sensorial responses over ring finger (RF).Materials and methodsResults of individuals admitted to our ENMG laboratory between June 2011 and March 2012 for nerve conduction studies (NCSs) were retrospectively analyzed. Sensory NCSs were performed by standard antidromic technique.ResultsTotally, 112 normal recordings belong to 100 patients were included. Mean antidromic sensory conduction velocity of MNs (wrist-to-second finger) or UNs (wrist-to-fifth finger) was not different between two genders. Mean sensory nerve action potential (SNAP) amplitude of MN from second finger was also not different between two genders. However, mean SNAP amplitude of UN from fifth finger was higher in females. In RF’s sensorial response studies; mean peak latency of MN was similar between females and males (3.05 ± 0.25 ms vs. 3.14 ± 0.29 ms, p = 0.111), whereas one of UN was shorter in females (2.86 ± 0.22 ms vs. 3.04 ± 0.31 ms, p = 0.001). MN to UN latency difference to RF was greater in females than males (0.19 ± 0.15 ms vs. 0.10 ± 0.16 ms, p = 0.007). Mean SNAP amplitude of MN and UN were both higher in females than males (17.9 ± 7.1 μV vs. 14.1 ± 5.5 μV, p = 0.011 and 18.5 ± 8.0 μV vs. 12.9 ± 6.1 μV, p = 0.0009, respectively). All data of NCSs were re-analyzed after adjustment for age, and obtained findings regarding effect of aging are also included.ConclusionGender has a prominent effect on RF’s sensorial responses. Normative values regarding them should be prepared with adjustment for gender.     

The architecture and contraction time of intrinsic foot muscles

Although critical for effective human locomotion and posture, little data exists regarding the segmentation, architecture and contraction time of the human intrinsic foot muscles. To address this issue, the Abductor Hallucis (AH), Abductor Digiti Minimi (ADM), Flexor Digitorum Brevis (FDB) and Extensor Digitorum Brevis (EDB) were investigated utilizing a cadaveric dissection and a non-invasive whole muscle mechanomyographic (wMMG) technique. The segmental structure and architecture of formaldehyde-fixed foot specimens were determined in nine cadavers aged 60–80 years. The wMMG technique was used to determine the contraction time (Tc) of individual muscle segments, within each intrinsic foot muscle, in 12 volunteers of both genders aged between 19 and 24 years.While the pattern of segmentation and segmental –architecture (e.g. fibre length) and –Tc of individual muscle segments within the same muscle were similar, they varied between muscles. Also, the average whole muscle Tc of FDB was significantly (p < 0.05) shorter (faster) (Tc = 58 ms) than in all other foot muscles investigated (ADM Tc = 72 ms, EDB Tc = 72 ms and ABH Tc = 69 ms). The results suggest that the architecture and contraction time of the FDB reflect its unique direct contribution, through toe flexion, to postural stability and the rapid development of ground reaction forces during forceful activities such as running and jumping.     

Algorithmic detection of the beginning and end of bipolar electrograms: Implications for novel methods to assess local activation time during atrial tachycardia

Activation mapping is required to effectively ablate atrial tachycardia (AT). Conventional tools to assess local activation time (LAT) are based upon the peak of the bipolar electrogram (B-EGM, LAT_Peak) and the maximal negative slope of the unipolar electrogram (U-EGM, LAT_Slope). Bipolar electrograms are influenced by wavefront direction, bipole orientation, and inter-electrode spacing causing ambiguity in peak detection, whereas unipolar electrograms are disturbed by the presence of far-field signals. We developed a new algorithm to detect the beginning and end of bipolar electrograms (t_begin and t_end). Then, we introduced new LAT methods related to the onset of B-EGMs (LAT_Onset), the center of mass of B-EGMs (LAT_CoM), and the slope of U-EGMs within a pre-defined window (LAT_Slope-hybrid).In total 3752 recordings from 31 AT patients were retrospectively analyzed. The signal-to-noise ratio (SNR) for B-EGMs was calculated to differentiate algorithmically high from low quality electrograms (HQ and LQ). In a subset of 328 B-EGMs, five experts validated the t_begin as determined by the algorithm by visual rating. The newly developed LAT methods were compared to the conventional LAT methods and to one another (Bland–Altman plots) in both HQ (n = 3003) and LQ EGMs (n = 749).The t_begin algorithm was accurate (deviation < ±10 ms) in 96 ± 4% of HQ and 91 ± 8% of LQ B-EGMs. BA plots revealed the following difference (bias) and variation in HQ and LQ EGMs respectively: (1) LAT_Onset vs. LAT_Peak: 27 ± 30 ms and 24 ± 62 ms; (2) LAT_CoM vs. LAT_Peak: 0 ± 16 ms and 2 ± 38 ms; (3) LAT_Slope-hybrid vs. LAT_Slope: 1 ± 32 ms and 15 ± 110 ms; (4) LAT_Onset vs. LAT_CoM: 22 ± 24 ms and 18 ± 22 ms; (5) LAT_Onset vs. LAT_Slope-hybrid: 16 ± 18 ms and 13 ± 22 ms; and (6) LAT_CoM vs. LAT_Slope-hybrid: 5 ± 20 ms and 4 ± 18 ms.In the present study, we introduced three new methods to assess local activation time in AT, based upon an algorithm detecting accurately the beginning and end of the B-EGM complex. BA analysis of the new methods showed similar variation in high and low quality EGMs, suggesting that they introduce less ambiguity than the conventional peak method. LAT_Onset consistently yielded an earlier activation moment. LAT_Slope-hybrid – by blanking far-field potentials – seems to be the optimal method for detection of the maximal negative slope in U-EGMs. Interestingly, LAT_CoM in B-EGMs coincided with the maximal negative slope in U-EGMs, suggesting its physiological sense and future use. The new LAT methods can be implemented in real-time mapping applications.     

Stability and activity of Dictyoglomus thermophilum GH11 xylanase and its disulphide mutant at high pressure and temperature

The functional properties of extremophilic Dictyoglomus thermophilum xylanase (XYNB) and the N-terminal disulphide-bridge mutant (XYNB-DS) were studied at high pressure and temperature. The enzymes were quite stable even at the pressure of 500 MPa at 80 °C. The half-life of inactivation in these conditions was over 30 h. The inactivation at 80 °C in atmospheric pressure was only 3-times slower. The increase of pressure up to 500 MPa at 80 °C decreased only slightly the enzyme's stability, whereas in 500 MPa the increase of temperature from 22 to 80 °C decreased significantly more the enzyme's stability. While the high temperature (80–100 °C) decreased the enzyme reaction with short xylooligosaccharides (xylotetraose and xylotriose), the high pressure (100–300 MPa) had an opposite effect. The temperature of 100 °C strongly increased the K_m but did not affect the k_cat to the same extent, thus indicating that the interaction of the substrate with the active site suffers before the catalytic reaction begins to decrease as the temperature rises. Circular dichroism spectroscopy showed the high structural stability of XYNB and XYNB-DS at 93 °C.     

Electromyographic latency of postural evoked responses from the leg muscles during EquiTest Computerised Dynamic Posturography: Reference data on healthy subjects

No normative data are available for the latencies of the EMG signals from the ankle muscles in response to sudden sagittal tilt (toes-UP or toes-DOWN) or shift (shift-FOR or shift-BACK) of the support surface during standing. In this study the postural evoked response (PER) paradigm on the EquiTest™ force platform was applied to 31 healthy adults (18 women and 13 men; mean age 29 years). The EMG latencies (PEREMG) were computed both through the standard manual procedure and through a specially designed automated algorithm. The manually computed PEREMG onset yielded a 95% tolerance interval between 82 ms and 148 ms after toes-UP perturbation, between 93 ms and 182 ms after toes-DOWN perturbation, between 67 ms and 107 ms after shift-BACK perturbation, and between 73 ms and 113 ms after shift-FOR perturbation. When comparing the two methods, paired t-tests showed no significant mean difference (Bonferroni-adjusted p-values ranged from 0.440 to 1.000) and all Bland–Altman plots included zero difference within the limits of agreement. Therefore, the manual and the automated methods appear to be sufficiently consistent. These results foster the clinical application of PEREMG testing on the EquiTest platform.     

The effect of PI3 kinase inhibitor LY294002 on voltage-dependent K+ channels in rabbit coronary arterial smooth muscle cells

AimsWe examined the effect of LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, on voltage-dependent K⁺ (Kv) channels.Main methodsElectrophysiological recordings were performed in freshly isolated rabbit coronary arterial smooth muscle cells.Key findingsThe Kv current amplitude was inhibited by LY294002 in a dose-dependent manner, with a K_d value of 1.48 μM. Without alteration of the kinetics of activation, LY294002 accelerated the decay rate of Kv channel inactivation. The rate constants of association and dissociation for LY294002 were 1.83 ± 0.01 μM^? 1 s^? 1 and 2.59 ± 0.14 s^? 1, respectively. Application of LY294002 had no significant impact on the steady-state activation or inactivation curves. In the presence of LY294002, the recovery time constant from inactivation was increased, and Kv channel inhibition increased under train pulses (1 or 2 Hz). This indicates that LY294002-induced Kv channel inhibition is use-dependent. Furthermore, pretreatment with another PI3K inhibitor, wortmannin (10 μM), did not affect the Kv current, and did not change the inhibitory effect of LY294002.SignificanceBased on these results, we suggest that LY294002 directly blocks Kv current irrespective of PI3K inhibition.     

Kinetic analysis for suicide-substrate inactivation of microperoxidase-11: A modified model for bisubstrate enzymes in the presence of reversible inhibitors

Kinetics of microperoxidase-11 (MP-11) as a heme–peptide enzyme model in oxidation reaction of guaiacol (AH) by hydrogen peroxide was studied in the presence of amino acids, taking into account the inactivation of MP-11 during reaction by its suicide substrate, H₂O₂. Reliability of the kinetic equation was evaluated by non-linear mathematical fitting. Fitting of experimental data into a new integrated kinetic relation showed a close match between the kinetic model and the experimental data. Indeed, it was found that the mechanism of suicide-peroxide inactivation of MP-11 in the presence of amino acids is different from MP-11 and/or horseradish peroxidase. In this mechanism, amino acids compete with hydrogen peroxide for the sixth co-ordination position of iron atom in the heme group through a competitive inhibition mechanism.The proposed model can successfully determine the kinetic parameters including inactivation by hydrogen peroxide as well as the inhibitory rate constants by the amino acid inhibitor.Kinetic parameters of inactivation including the initial activity of MP-11, α₀, the apparent inactivation rate constant, k_i and the apparent inhibition rate constant for cysteine, k_I were obtained 0.282 ± 0.006 min^?1, 0.497 ± 0.013 min^?1 and 1.374 ± 0.007 min^?1 at [H₂O₂] = 1.0 mM, 27 °C, phosphate buffer 5.0 mM, pH 7.0. Results showed that inactivation and inhibition of microperoxidase as a peroxidase model enzyme occurred simultaneously even at low concentrations of hydrogen peroxide (0.4 mM). This kinetic analysis based on the suicide-substrate inactivation of microperoxidase-11, provides a tool and model for studying peroxidase models in the presence of reversible inhibitors. The introduced inhibition procedure can be used in designing activity tunable and specific protected enzyme models in the hidden and reversibly inhibited forms, which do not undergo inactivation.     

Signal-to-noise ratio of diffusion weighted magnetic resonance imaging: Estimation methods and in vivo application to spinal cord

Diffusion tensor imaging (DTI) and tractographic reconstruction may be applied for in vivo clinical spinal cord studies. However, this structure represents a challenge to current acquisition and reconstruction strategies, due to its small size, motion artifacts, partial volume effects and low signal-to-noise-ratio (SNR). Aims of this work were to select the best approach for the estimate of SNR and to use it for spinal cord diffusion weighted (DW) sequence optimization.Seven methods for the estimate of SNR were compared on uniform phantom DW images, and the best performing approach (single ROI for signal and noise, difference of images—SNR_diff) was applied for the following in vivo sequence evaluations.Fifteen sequences with different parameters (voxel size, repetition (TR) and echo (TE) times) were compared according to SNR, resolution, fractional anisotropy (FA) and tractography performances on three healthy volunteers. In vivo optimization of DW sequences resulted in: axial sequence, with voxel size = 1.5 mm × 1.5 mm × 3.5 mm, TR = 3200 ms and TE = 89 ms, sagittal sequence with voxel size = 2.2 mm × 2.2 mm × 2 mm, TR = 3000 ms and TE = 84 ms.An objective method tested on phantom and a practical index for in vivo spinal cord DTI SNR estimation allowed to obtain axial and sagittal optimized sequences, providing excellent tractographic results, with acceptable acquisition times for in vivo clinical applications.     

Expanded bed adsorption of an alkaline lipase from Pseudomona cepacia

An extracellular lipase was isolated from Pseudomona cepacia by expanded bed adsorption on an Amberlite 410 ion-exchange resin. Enzyme characterization and hydrodynamic study of a chromatography column were done. Enzyme purification was done at three condition of expanded bed height (H): at one and half (6 cm), at two (8 cm) and at three (12 cm) times the fixed bed height (H₀ = 4 cm). The results showed that the experimental data was fitted to the Richardson and Zaki equation, and the comparison between the experimental and calculated terminal velocities showed low relative error. In enzyme purification for better condition, a purification factor of about 80 times was found at 6 cm of expanded bed height, or 1.5 times of expansion degree. Purified lipase had an optimal pH and a temperature of 8 and 37 °C, respectively.