Flexible array coil for cervical and extraspinal (FACE) MRI at 3.0 Tesla.

Objective.High-resolution MRI of the cervical spine (c-spine) and extraspinal neck region requires close-fitting receiver coils to maximize the signal-to-noise ratio (SNR). Conventional, rigid C-spine receiver coils do not adequately contour to the neck to accommodate varying body shapes, resulting in suboptimal SNR. Recent innovations in flexible surface coil array designs may provide three-dimensional (3D) bendability and conformability to optimize SNR, while improving capabilities for higher acceleration factors.Approach.This work describes the design, implementation, and preliminaryin vivotesting of a novel, conformal 23-channel receive-only flexible array for cervical and extraspinal (FACE) MRI at 3-Tesla (T), with use of high-impedance elements to enhance the coil's flexibility. Coil performance was tested by assessing SNR and geometry factors (g-factors) in a phantom compared to a conventional 21-channel head-neck-unit (HNU).In vivoimaging was performed in healthy human volunteers and patients using high-resolution c-spine and neck MRI protocols at 3T, including MR neurography (MRN).Main results.Mean SNR with the FACE was 141%-161% higher at left, right, and posterior off-isocenter positions and 4% higher at the isocenter of the phantom compared to the HNU. Parallel imaging performance was comparable for an acceleration factor (R) = 2 × 2 between the two coils, but improved forR= 3 × 3 with meang-factors ranging from 1.46-2.15 with the FACE compared to 2.36-3.62 obtained with the HNU. Preliminary human volunteer and patient testing confirmed that equivalent or superior image quality could be obtained for evaluation of osseous and soft tissue structures of the cervical region with the FACE.Significance.A conformal and highly flexible cervical array with high-impedance coil elements can potentially enable higher-resolution imaging for cervical imaging.

Post-exercise intramuscular O2 supply is tightly coupled with a higher proximal-to-distal ATP synthesis rate in human tibialis anterior.

KEY POINTS: The post-exercise recovery of phosphocreatine, a measure of the oxidative capacity of muscles, as assessed by 31 P MR spectroscopy, shows a striking increase from distal to proximal along the human tibialis anterior muscle. To investigate why this muscle exhibits a greater oxidative capacity proximally, we tested whether the spatial variation in phosphocreatine recovery rate is related to oxygen supply, muscle fibre type or type of exercise. We revealed that oxygen supply also increases from distal to proximal along the tibialis anterior, and that it strongly correlated with phosphocreatine recovery. Carnosine level, a surrogate measure for muscle fibre type was not different between proximal and distal, and type of exercise did not affect the gradient in phosphocreatine recovery rate. Taken together, the findings of this study suggest that the post-exercise spatial gradients in oxygen supply and phosphocreatine recovery are driven by a higher intrinsic mitochondrial oxidative capacity proximally. ABSTRACT: Phosphorus magnetic resonance spectroscopy (31 P MRS) of human tibialis anterior (TA) revealed a strong proximo-distal gradient in the post-exercise phosphocreatine (PCr) recovery rate constant (kPCr ), a measure of muscle oxidative capacity. The aim of this study was to investigate whether this kPCr gradient is related to O2 supply, resting phosphorylation potential, muscle fibre type, or type of exercise. Fifteen male volunteers performed continuous isometric ankle dorsiflexion at 30% maximum force until exhaustion. At multiple locations along the TA, we measured the oxidative PCr resynthesis rate (VPCr = kPCr × PCr depletion) by 31 P MRS, the oxyhaemoglobin recovery rate constant (kO2Hb ) by near infrared spectroscopy, and muscle perfusion with MR intravoxel incoherent motion imaging. The kO2Hb , kPCr , VPCr and muscle perfusion depended on measurement location (P < 0.001, P < 0.001, P = 0.032 and P = 0.003, respectively), all being greater proximally. The kO2Hb and muscle perfusion correlated with kPCr (r = 0.956 and r = 0.852, respectively) and VPCr (r = 0.932 and r = 0.985, respectively), the latter reflecting metabolic O2 consumption. Resting phosphorylation potential (PCr/inorganic phosphate) was also higher proximally (P < 0.001). The surrogate for fibre type, carnosine content measured by 1 H MRS, did not differ between distal and proximal TA (P = 0.884). Performing intermittent exercise to avoid exercise ischaemia, still led to larger kPCr proximally than distally (P = 0.013). In conclusion, the spatial kPCr gradient is strongly associated with the spatial variation in O2 supply. It cannot be explained by exercise-induced ischaemia nor by fibre type. Our findings suggest it is driven by a higher proximal intrinsic mitochondrial oxidative capacity, apparently to support contractile performance of the TA.

3D 31 P MR spectroscopic imaging of the human brain at 3 T with a 31 P receive array: An assessment of 1 H decoupling, T1 relaxation times, 1 H-31 P nuclear Overhauser effects and NAD.

31 P MR spectroscopic imaging (MRSI) is a versatile technique to study phospholipid precursors and energy metabolism in the healthy and diseased human brain. However, mainly due to its low sensitivity, 31 P MRSI is currently limited to research purposes. To obtain 3D 31 P MRSI spectra with improved signal-to-noise ratio on clinical 3 T MR systems, we used a coil combination consisting of a dual-tuned birdcage transmit coil and a 31 P eight-channel phased-array receive insert. To further increase resolution and sensitivity we applied WALTZ4 1 H decoupling and continuous wave nuclear Overhauser effect (NOE) enhancement and acquired high-quality MRSI spectra with nominal voxel volumes of ~ 17.6 cm3 (effective voxel volume ~ 51 cm3 ) in a clinically relevant measurement time of ~ 13 minutes, without exceeding SAR limits. Steady-state NOE enhancements ranged from 15 ± 9% (γ-ATP) and 33 ± 3% (phosphocreatine) to 48 ± 11% (phosphoethanolamine). Because of these improvements, we resolved and detected all 31 P signals of metabolites that have also been reported for ultrahigh field strengths, including resonances for NAD+ , NADH and extracellular inorganic phosphate. T1 times of extracellular inorganic phosphate were longer than for intracellular inorganic phosphate (3.8 ± 1.4s vs 1.8 ± 0.65 seconds). A comparison of measured T1 relaxation times and NOE enhancements at 3 T with published values between 1.5 and 9.4 T indicates that T1 relaxation of 31 P metabolite spins in the human brain is dominated by dipolar relaxation for this field strength range. Even although intrinsic sensitivity is higher at ultrahigh fields, we demonstrate that at a clinical field strength of 3 T, similar 31 P MRSI information content can be obtained using a sophisticated coil design combined with 1 H decoupling and NOE enhancement.

A multitransmit external body array combined with a 1 H and 31 P endorectal coil to enable a multiparametric and multimetabolic MRI examination of the prostate at 7T.

PURPOSE: In vivo 1 H and 31 P magnetic resonance spectroscopic imaging (MRSI) provide complementary information on the biology of prostate cancer. In this work we demonstrate the feasibility of performing multiparametric imaging (mpMRI) and 1 H and 31 P spectroscopic imaging of the prostate using a 31 P and 1 H endorectal radiofrequency coil (ERC) in combination with a multitransmit body array at 7 Tesla (T). METHODS: An ERC with a 31 P transceiver loop coil and 1 H receive (Rx) asymmetric microstrip (31 P/1 H ERC) was designed, constructed and tested in combination with an external 8-channel 1 H transceiver body array coil (8CH). Electromagnetic field simulations and measurements and in vivo temperature measurements of the ERC were performed for safety validation. In addition, the signal-to-noise (SNR) benefit of the 1 H microstrip with respect to the 8CH was evaluated. Finally, the feasibility of the setup was tested in one volunteer and three patients with prostate cancer by performing T2 -weighted and diffusion-weighted imaging in combination with 1 H and 31 P spectroscopic imaging. RESULTS: Electromagnetic field simulations of the 31 P loop coil showed no differences in the E- and B-fields of the 31 P/1 H ERC compared with a previously safety validated ERC without 1 H microstrip. The hotspot of the specific absorption rate (SAR) at the feed point of the 31 P/1 H ERC loop coil was 9.42 W/kg when transmitting on 31 P at 1 W. Additional in vivo measurements showed a maximum temperature increase at the SAR hotspot of 0.7°C over 6 min on 31 P at 1.9 W transmit (Tx) power, indicating safe maximum power levels. When transmitting with the external 1 H body array at 40W for 2:30 min, the temperature increase around the ERC was < 0.3°C. Up to 3.5 cm into the prostate the 1 H microstrip of the ERC provided higher SNR than the 8CH. The total coil combination allowed acquisition of an mpMRI protocol and the assessment of 31 P and 1 H metabolites of the prostate in all test subjects. CONCLUSION: We developed a setup with a 31 P transceiver and 1 H Rx endorectal coil in combination with an 8-channel transceiver external body array coil and demonstrated its safety and feasibility for obtaining multiparametric imaging and 1 H and 31 P MRSI at 7T in patients with prostate cancer within one MR examination.

An 8-channel receive array for improved 31 P MRSI of the whole brain at 3T.

PURPOSE: To demonstrate a 1 H/31 P whole human brain volume coil configuration for 3 Tesla with separate 31 P transmit and receive components that maintains 1 H MRS performance and delivers optimal 31 P MRSI with 1 H decoupling. METHODS: We developed an 8-channel 31 P receive array coil covering the head to be used as an insert for a commercial double-tuned 1 H/31 P birdcage transmit-receive coil. This retains the possibility of using low-power rectangular pulses for 1 H-decoupled 3D 31 P MRSI (nominal resolution 17.6 cm3 ; acquisition duration 13 min) but increases the SNR with the receive sensitivity of 31 P surface coils. The performance of the combined coil setup was evaluated by measuring 1 H and 31 P SNR with and without the 31 P receive array and by assessing the effect of the receive array on the transmit efficiencies of the birdcage coil. RESULTS: Compared to the birdcage coil alone, the 31 P insert in combination with the birdcage achieved an average 31 P SNR gain of 1.4 ± 0.4 in a center partition of the brain. The insert did not cause losses in 1 H MRS performance and transmit efficiency, whereas for 31 P approximately 20% more power was needed to achieve the same γB1. CONCLUSION: The new coil configuration allows 1 H MRSI and optimal 1 H-decoupled 3D 31 P MRSI, with increased SNR of the human brain without patient repositioning, for clinical and research purposes at 3 Tesla.

Evaluation of fluorescent dyes to measure protein aggregation within mammalian cell culture supernatants.

BACKGROUND: A current challenge in bioprocessing is the ability to analyse critical quality attributes such as aggregation without prior purification. This study evaluated the use of fluorescent dyes (Bis-ANS, SYPRO Orange, Thioflavin T and ProteoStat) to characterise mAb aggregates in Chinese hamster ovary clarified cultures. RESULTS: The null and mAb culture supernatants showed an increase in fluorescence intensity over the duration of the culture. The null cultures on day 14 saw a rapid increase in fluorescence intensity; day 10 to day 14, Bis-ANS and Thioflavin T had average increases of 21% and 48%, respectively, whereas ProteoStat and SYPRO Orange showed an average increase of 60%. Higher fluorescence intensity on day 14 with the null cultures, also correlated with loss of viability. CONCLUSION: Fluorescent dyes are not a specific indicator of mAb aggregation, but rather an indicator of overall protein aggregation or high molecular weight species. SYPRO Orange was more sensitive at detecting very large molecular weight species and ProteoStat seemed better suited to smaller aggregates. Although the assay cannot be used to measure mAb aggregates in cell culture, it could be used to aid cell line selection in maximising viabilities and minimising the amount of aggregates. © 2017 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Oxidative capacity varies along the length of healthy human tibialis anterior.

KEY POINTS: During exercise skeletal muscles use the energy buffer phosphocreatine. The post-exercise recovery of phosphocreatine is a measure of the oxidative capacity of muscles and is traditionally assessed by 31 P magnetic resonance spectroscopy of a large tissue region, assuming homogeneous energy metabolism. To test this assumption, we collected spatially resolved spectra along the length of human tibialis anterior using a home-built array of 31 P detection coils, and observed a striking gradient in the recovery rate of phosphocreatine, decreasing along the proximo-distal axis of the muscle. A similar gradient along this muscle was observed in signal changes recorded by 1 H muscle functional MRI. These findings identify intra-muscular variation in the physiology of muscles in action and highlight the importance of localized sampling for any methodology investigating oxidative metabolism of this, and potentially other muscles. ABSTRACT: The rate of phosphocreatine (PCr) recovery (kPCr ) after exercise, characterizing muscle oxidative capacity, is traditionally assessed with unlocalized 31 P magnetic resonance spectroscopy (MRS) using a single surface coil. However, because of intramuscular variation in fibre type and oxygen supply, kPCr may be non-uniform within muscles. We tested this along the length of the tibialis anterior (TA) muscle in 10 male volunteers. For this purpose, we employed a 3T MR system with a 31 P/1 H volume transmit coil combined with a home-built 31 P phased-array receive probe, consisting of five coil elements covering the TA muscle length. Mono-exponential kPCr was determined for all coil elements after 40 s of submaximal isometric dorsiflexion (SUBMAX) and incremental exercise to exhaustion (EXH). In addition, muscle functional MRI (1 H mfMRI) was performed using the volume coil after another 40 s of SUBMAX. A strong gradient in kPCr was observed along the TA (P < 0.001), being two times higher proximally vs. distally during SUBMAX and EXH. Statistical analysis showed that this gradient cannot be explained by pH variations. A similar gradient was seen in the slope of the initial post-exercise 1 H mfMRI signal change, which was higher proximally than distally in both the TA and the extensor digitorum longus (P < 0.001) and strongly correlated with kPCr . The pronounced differences along the TA in functional oxidative capacity identify regional variation in the physiological demand of this muscle during everyday activities and have implications for the bio-energetic assessment of interventions to modify its performance and of neuromuscular disorders involving the TA.

Feasibility of Multiparametric Magnetic Resonance Imaging of the Prostate at 7 T.

OBJECTIVES: The aim of this study was to evaluate the technical feasibility of prostate multiparametric magnetic resonance imaging (mpMRI) at a magnetic field strength of 7 T. MATERIALS AND METHODS: In this prospective institutional review board-approved study, 14 patients with biopsy-proven prostate cancer (mean age, 65.2 years; median prostate-specific antigen [PSA], 6.2 ng/mL), all providing signed informed consent, underwent 7 T mpMRI with an external 8-channel body-array transmit coil and an endorectal receive coil between September 2013 and October 2014. Image and spectral quality of high-resolution T2-weighted (T2W) imaging (0.3 × 0.3 × 2 mm), diffusion-weighted imaging (DWI; 1.4 × 1.4 × 2 mm or 1.75 × 1.75 × 2 mm), and (H) MR spectroscopic imaging (MRSI; real voxel size, 0.6 mm in 7:16 minutes) were rated on a 5-point scale by 2 radiologists and a spectroscopist. RESULTS: Prostate mpMRI including at least 2 of 3 MR techniques was obtained at 7 T in 13 patients in 65 ± 12 minutes. Overall T2W and DWI image quality at 7 T was scored as fair (38% and 17%, respectively) to good or very good (55% and 83%, respectively). The main artifacts for T2W imaging were motion and areas of low signal-to-noise ratio, the latter possibly caused by radiofrequency field inhomogeneities. For DWI, the primary artifact was ghosting of the rectal wall in the readout direction. Magnetic resonance spectroscopic imaging quality was rated fair or good in 56% of the acquisitions and was mainly limited by lipid contamination. CONCLUSIONS: Multiparametric MRI of the prostate at 7 T is feasible at unprecedented spatial resolutions for T2W imaging and DWI and within clinically acceptable acquisition times for high-resolution MRSI, using the combination of an external 8-channel body-array transmit coil and an endorectal receive coil. The higher spatial resolutions can yield improved delineation of prostate anatomy, but the robustness of the techniques needs to be improved before clinical adoption of 7 T mpMRI.

Evaluation of options for harvest of a recombinant E. Coli fermentation producing a domain antibody using ultra scale-down techniques and pilot-scale verification.

Ultra scale-down (USD) methods operating at the millilitre scale were used to characterise full-scale processing of E. coli fermentation broths autolysed to different extents for release of a domain antibody. The focus was on the primary clarification stages involving continuous centrifugation followed by depth filtration. The performance of this sequence was predicted by USD studies to decrease significantly with increased extents of cell lysis. The use of polyethyleneimine reagent was studied to treat the lysed cell broth by precipitation of soluble contaminants such as DNA and flocculation of cell debris material. The USD studies were used to predict the impact of this treatment on the performance and here it was found that the fermentation could be run to maximum productivity using an acceptable clarification process (e.g., a centrifugation stage operating at 0.11 L/m(2) equivalent gravity settling area per hour followed by a resultant required depth filter area of 0.07 m(2) /L supernatant). A range of USD predictions was verified at the pilot scale for centrifugation followed by depth filtration. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:382-392, 2016.

(1)H MR spectroscopic imaging of the prostate at 7T using spectral-spatial pulses.

PURPOSE: To assess the feasibility of prostate (1)H MR spectroscopic imaging (MRSI) using low-power spectral-spatial (SPSP) pulses at 7T, exploiting accurate spectral selection and spatial selectivity simultaneously. METHODS: A double spin-echo sequence was equipped with SPSP refocusing pulses with a spectral selectivity of 1 ppm. Three-dimensional prostate (1)H-MRSI at 7T was performed with the SPSP-MRSI sequence using an 8-channel transmit array coil and an endorectal receive coil in three patients with prostate cancer and in one healthy subject. No additional water or lipid suppression pulses were used. RESULTS: Prostate (1)H-MRSI could be obtained well within specific absorption rate (SAR) limits in a clinically feasible time (10 min). Next to the common citrate signals, the prostate spectra exhibited high spermine signals concealing creatine and sometimes also choline. Residual lipid signals were observed at the edges of the prostate because of limitations in spectral and spatial selectivity. CONCLUSION: It is possible to perform prostate (1)H-MRSI at 7T with a SPSP-MRSI sequence while using separate transmit and receive coils. This low-SAR MRSI concept provides the opportunity to increase spatial resolution of MRSI within reasonable scan times.

Enhancing the selective extracellular location of a recombinant E. coli domain antibody by management of fermentation conditions.

The preparation of a recombinant protein using Escherichia coli often involves a challenging primary recovery sequence. This is due to the inability to secrete the protein to the extracellular space without a significant degree of cell lysis. This results in the release of nucleic acids, leading to a high viscosity, difficulty to clarify, broth and also to contamination with cell materials such as lipopolysaccharides and host cell proteins. In this paper, we present different fermentation strategies to facilitate the recovery of a V H domain antibody (13.1 kDa) by directing it selectively to the extracellular space and changing the balance between domain antibody to nucleic acid release. The manipulation of the cell growth rate in order to increase the outer cell membrane permeability gave a small ~1.5-fold improvement in released domain antibody to nucleic acid ratio without overall loss of yield. The introduction during fermentation of release agents such as EDTA gave no improvement in the ratio of released domain antibody to nucleic acid and a loss of overall productivity. The use of polyethyleneimine (PEI) during fermentation was with the aim to (a) permeabilise the outer bacterial membrane to release selectively domain antibody and (b) remove selectively by precipitation nucleic acids released during cell lysis. This strategy resulted in up to ~4-fold increase in the ratio of domain antibody to soluble nucleic acid with no reduction in domain antibody overall titre. In addition, a reduction in host cell protein contamination was achieved and there was no increase in endotoxin levels. Similar results were demonstrated with a range of other antibody products prepared in E. coli.

Cell tracking using (19)F magnetic resonance imaging: technical aspects and challenges towards clinical applications.

(19)F MRI is emerging as a new imaging technique for cell tracking. It is particularly attractive because of its potential for direct and precise cell quantification. The most important challenge towards in vivo applications is the sensitivity of the technique, i.e. the detection limit in a reasonable imaging time. Optimal sensitivity can be achieved with dedicated (19)F compounds together with specifically adapted hardware and acquisition methods. In this paper we introduce the (19)F MRI technique focusing on these key sensitivity issues and review the state-of-the-art of (19)F MRI and developments towards its clinical use. We calculate (19)F detection limits reported in preclinical cell and clinical (19)F drug studies in terms of tissue concentration in a 1 cm(3) voxel, as an alternate way to compare detection limits. We estimate that a tissue concentration of a few millimoles per litre (mM) of (19)F is required for a human study at a resolution of 1 cm(3).

(31) P MR spectroscopic imaging of the human prostate at 7 T: T1 relaxation times, Nuclear Overhauser Effect, and spectral characterization.

PURPOSE: Optimization of phosphorus ((31) P) MR spectroscopic imaging (MRSI) of the human prostate at 7 T by the evaluation of T1 relaxation times and the Nuclear Overhauser Effect (NOE) of phosphorus-containing metabolites. METHODS: Twelve patients with prostate cancer and one healthy volunteer were scanned on a 7 T whole-body system using a (31) P endorectal coil combined with an eight-channel (1) H body array coil. T1 relaxation times were measured using progressive saturation in a two-dimensional localization sequence. (31) P MRSI was performed twice: once without NOE and once with NOE using low-power continuous wave (1) H irradiation to determine NOE enhancements. RESULTS: T1 relaxation times of (31) P metabolites in the human prostate at 7 T varied between 3.0 and 8.3 s. Positive but variable NOE enhancements were measured for most metabolites. Remarkably, the (31) P MR spectra showed two peaks in chemical shift range of inorganic phosphate. CONCLUSION: Knowledge of T1 relaxation times and NOE enhancements enables protocol optimization for (31) P MRSI of the prostate at 7 T. With a strongly reduced (31) P flip angle (≤ 45°), a (31) P MRSI dataset with optimal signal-to-noise ratio per unit time can be obtained within 15 minutes. The NOE enhancement can improve fitting accuracy, but its variability requires further investigation.

Clinical comparison between a currently available single-loop and an investigational dual-channel endorectal receive coil for prostate magnetic resonance imaging: a feasibility study at 1.5 and 3 T.

OBJECTIVES: The objectives of this study were to test the feasibility of an investigational dual-channel next-generation endorectal coil (NG-ERC) in vivo, to quantitatively assess signal-to-noise ratio (SNR), and to get an impression of image quality compared with the current clinically available single-loop endorectal coil (ERC) for prostate magnetic resonance imaging at both 1.5 and 3 T. MATERIALS AND METHODS: The study was approved by the institutional review board, and written informed consent was obtained from all patients. In total, 8 consecutive patients with prostate cancer underwent a local staging magnetic resonance examination with the successive use of both coils in 1 session (4 patients at 1.5 T and 4 other patients at 3 T). Quantitative comparison of both coils was performed for the apex, mid-gland and base levels at both field strengths by calculating SNR profiles in the axial plane on an imaginary line in the anteroposterior direction perpendicular to the coil surface. Two radiologists independently assessed the image quality of the T2-weighted and apparent diffusion coefficient maps calculated from diffusion-weighted imaging using a 5-point scale. Improvement of geometric distortion on diffusion-weighted imaging with the use of parallel imaging was explored. Statistical analysis included a paired Wilcoxon signed rank test for SNR and image quality evaluation as well as κ statistics for interobserver agreement. RESULTS: No adverse events were reported. The SNR was higher for the NG-ERC compared with the ERC up to a distance of approximately 40 mm from the surface of the coil at 1.5 T (P < 0.0001 for the apex, the mid-gland, and the base) and approximately 17 mm (P = 0.015 at the apex level) and 30 mm at 3 T (P < 0.0001 for the mid-gland and base). Beyond this distance, the SNR profiles of both coils were comparable. Overall, T2-weighted image quality was considered better for NG-ERC at both field strengths. Quality of apparent diffusion coefficient maps with the use of parallel imaging was rated superior with the NG-ERC at 3 T. CONCLUSIONS: The investigational NG-ERC for prostate imaging outperforms the current clinically available ERC in terms of SNR and is feasible for continued development for future use as the next generation endorectal coil for prostate imaging in clinical practice.

Phosphorus magnetic resonance spectroscopic imaging at 7 T in patients with prostate cancer.

OBJECTIVES: The aim of this study was to identify characteristics of phosphorus (P) spectra of the human prostate and to investigate changes of individual phospholipid metabolites in prostate cancer through in vivo P magnetic resonance spectroscopic imaging (MRSI) at 7 T. MATERIALS AND METHODS: In this institutional review board-approved study, 15 patients with biopsy-proven prostate cancer underwent T2-weighted magnetic resonance imaging and 3-dimensional P MRSI at 7 T. Voxels were selected at the tumor location, in normal-appearing peripheral zone tissue, normal-appearing transition zone tissue, and in the base of the prostate close to the seminal vesicles. Phosphorus metabolite ratios were determined and compared between tissue types. RESULTS: Signals of phosphoethanolamine (PE) and phosphocholine (PC) were present and well resolved in most P spectra in the prostate. Glycerophosphocholine signals were observable in 43% of the voxels in malignant tissue, but in only 10% of the voxels in normal-appearing tissue away from the seminal vesicles. In many spectra, independent of tissue type, 2 peaks resonated in the chemical shift range of inorganic phosphate, possibly representing 2 separate pH compartments. The PC/PE ratio in the seminal vesicles was highly elevated compared with the prostate in 5 patients. A considerable overlap of P metabolite ratios was found between prostate cancer and normal-appearing prostate tissue, preventing direct discrimination of these tissues. The only 2 patients with high Gleason scores tumors (≥4+5) presented with high PC and glycerophosphocholine levels in their cancer lesions. CONCLUSIONS: Phosphorus MRSI at 7 T shows distinct features of phospholipid metabolites in the prostate gland and its surrounding structures. In this exploratory study, no differences in P metabolite ratios were observed between prostate cancer and normal-appearing prostate tissue possibly because of the partial volume effects of small tumor foci in large MRSI voxels.

In vivo 31P MR spectroscopic imaging of the human prostate at 7 T: safety and feasibility.

(31)P MR spectroscopic imaging of the human prostate provides information about phosphorylated metabolites that could be used for prostate cancer characterization. The sensitivity of a magnetic field strength of 7 T might enable 3D (31)P MR spectroscopic imaging with relevant spatial resolution in a clinically acceptable measurement time. To this end, a (31)P endorectal coil was developed and combined with an eight-channel (1)H body-array coil to relate metabolic information to anatomical location. An extensive safety validation was performed to evaluate the specific absorption rate, the radiofrequency field distribution, and the temperature distribution of both coils. This validation consisted of detailed Finite Integration Technique simulations, confirmed by MR thermometry and B 1+ measurements in a phantom and in vivo temperature measurements. The safety studies demonstrated that the presence of the (31)P endorectal coil had no influence on the specific absorption rate levels and temperature distribution of the external eight-channel (1)H array coil. To stay within a 10 g averaged local specific absorption rate of 10 W/kg, a maximum time-averaged input power of 33 W for the (1)H array coil was allowed. For transmitting with the (31)P endorectal coil, our safety limit of less than 1°C temperature increase in vivo during a 15-min MR spectroscopic imaging experiment was reached at a time-averaged input power of 1.9 W. With this power setting, a second in vivo measurement was performed on a healthy volunteer. Using adiabatic excitation, 3D (31)P MR spectroscopic imaging produced spectra from the entire prostate in 18 min with a spatial resolution of 4 cm(3). The spectral resolution enabled the separate detection of phosphocholine, phosphoethanolamine, inorganic phosphate, and other metabolites that could play an important role in the characterization of prostate cancer.

Optimised mammalian expression through the coupling of codon adaptation with gene amplification: maximum yields with minimum effort.

Gene amplification methodologies are frequently employed for the generation of large quantities of recombinant proteins in mammalian cells. Although they usually guarantee very high yields, they are very time consuming. In addition, due to the large genomic re-arrangements that frequently occur with amplification, the resulting high-producing clones can be unstable. We herein describe significant improvements to the dihydrofolate reductase (DHFR)/methotrexate (MTX) based gene amplification methodology typically employed to improve yields of recombinant proteins produced in genetically engineered CHO host cells. We demonstrate substantial synergy when such gene amplification is combined with extremely high codon optimisation strategies. As a result, expression saturation can be achieved rapidly, in as low as 5 nM MTX, with minimal effort and without compromise in final yields achieved.

Effective and robust plasmid topology analysis and the subsequent characterization of the plasmid isoforms thereby observed.

Within the biopharmaceutical industry, recombinant plasmid DNA is used both as a raw material (e.g. in lentiviral and AAV vector production) as well as an active ingredient (e.g. in DNA vaccines). Consequently, many analytical laboratories are routinely involved with plasmid DNA topoisoform qualitative analysis and quantification. In order to reliably determine plasmid topology, one must ensure that the methodology employed can reliably, precisely and accurately measure qualitatively and quantitatively all topological isoforms. Presented here are an anion-exchange high-performance liquid chromatography (AEC) and an agarose gel electrophoresis (AGE)-based method developed for this purpose. The strategies undertaken to overcome the respective typical problems of limited linear range of quantitation (for AGE) and isoform resolution (for AEC) are described. Also presented is a subsequent direct comparison (for assay precision/accuracy) of these two methods, as well as a package of species characterization [by chloroquine-AGE, enzymatic digestion, multi-angle laser light-scattering (MALLS) and electron microscopy] undertaken to confirm the identity of a minor supercoiled dimeric concatamer observed by both approaches.