• (18)F-VC701-PET and MRI in the in vivo neuroinflammation assessment of a mouse model of multiple sclerosis

      Belloli, Sara; Zanotti, Lucia; Murtaj, Valentina; Mazzon, Cristina; Di Grigoli, Giuseppe; Monterisi, Cristina; Masiello, Valeria; Iaccarino, Leonardo; Cappelli, Andrea; Poliani, Pietro Luigi; et al. (2018-02-05)
      BACKGROUND: Positron emission tomography (PET) using translocator protein (TSPO) ligands has been used to detect neuroinflammatory processes in neurological disorders, including multiple sclerosis (MS). The aim of this study was to evaluate neuroinflammation in a mouse MS model (EAE) using TSPO-PET with (18)F-VC701, in combination with magnetic resonance imaging (MRI). METHODS: MOG35-55/CFA and pertussis toxin protocol was used to induce EAE in C57BL/6 mice. Disease progression was monitored daily, whereas MRI evaluation was performed at 1, 2, and 4 weeks post-induction. Microglia activation was assessed in vivo by (18)F-VC701 PET at the time of maximum disease score and validated by radioligand ex vivo distribution and immunohistochemistry at 2 and 4 weeks post-immunization. RESULTS: In vivo and ex vivo analyses show that (18)F-VC701 significantly accumulates within the central nervous system (CNS), particularly in the cortex, striatum, hippocampus, cerebellum, and cervical spinal cord of EAE compared to control mice, at 2 weeks post-immunization. MRI confirmed the presence of focal brain lesions at 2 weeks post-immunization in both T1-weighted and T2 images. Of note, MRI abnormalities attenuated in later post-immunization phase. Neuropathological analysis confirmed the presence of microglial activation in EAE mice, consistent with the in vivo increase of (18)F-VC701 uptake. CONCLUSION: Increase of (18)F-VC701 uptake in EAE mice is strongly associated with the presence of microglia activation in the acute phase of the disease. The combined use of TSPO-PET and MRI provided complementary evidence on the ongoing disease process, thus representing an attractive new tool to investigate neuronal damage and neuroinflammation at preclinical levels.
    • 19F MRI Imaging of Polymer Nanogels Aided by Improved Segmental Mobility of Embedded Fluorine Moieties

      Munkhbat, Oyuntuya; Canakci, Mine; Zheng, Shaokuan; Hu, Weiguo; Osborne, Barbara; Bogdanov, Alexei A.; Thayumanavan, S. (2018-12-18)
      Using fluorinated probes for 19F MRI imaging is an emerging field with potential utility in cellular imaging and cell tracking in vivo, which complements conventional 1H MRI. An attractive feature of 19F-based imaging is that this is a bio-orthogonal nucleus and the naturally abundant isotope is NMR active. A significant hurdle however in the 19F MRI arises from the tendency of organic macromolecules, with multiple fluorocarbon substitutions, to aggregate in the aqueous phase. This aggregation results in significant loss of sensitivity, because the T2 relaxation times of these aggregated 19F species tend to be significantly lower. In this report, we have developed a strategy to covalently trap nanoscopic states with an optimal degree of 19F substitutions, followed by significant enhancement in T2 relaxation times through increased segmental mobility of the side chain substituents facilitated using stimulus-responsive elements in the polymeric nanogel. In addition to NMR relaxation time based evaluations, the ability to obtain such signals are also evaluated in mouse models. The propensity of these nanoscale assemblies to encapsulate hydrophobic drug molecules and the availability of surfaces for convenient introduction of fluorescent labels suggest the potential of these nanoscale architectures for use in multi-modal imaging and therapeutic applications.
    • 4-D Reconstruction With Respiratory Correction for Gated Myocardial Perfusion SPECT

      Qi, Wenyuan; Yang, Yongyi; Song, Chao; Wernick, Miles N.; Pretorius, P. Hendrik; King, Michael A. (2017-08-01)
      Cardiac single photon emission computed tomography (SPECT) images are known to suffer from both cardiac and respiratory motion blur. In this paper, we investigate a 4-D reconstruction approach to suppress the effect of respiratory motion in gated cardiac SPECT imaging. In this approach, the sequence of cardiac gated images is reconstructed with respect to a reference respiratory amplitude bin in the respiratory cycle. To combat the challenge of inherent high-imaging noise, we utilize the data counts acquired during the entire respiratory cycle by making use of a motion-compensated scheme, in which both cardiac motion and respiratory motion are taken into account. In the experiments, we first use Monte Carlo simulated imaging data, wherein the ground truth is known for quantitative comparison. We then demonstrate the proposed approach on eight sets of clinical acquisitions, in which the subjects exhibit different degrees of respiratory motion blur. The quantitative evaluation results show that the 4-D reconstruction with respiratory correction could effectively reduce the effect of motion blur and lead to a more accurate reconstruction of the myocardium. The mean-squared error of the myocardium is reduced by 22%, and the left ventricle (LV) resolution is improved by 21%. Such improvement is also demonstrated with the clinical acquisitions, where the motion blur is markedly improved in the reconstructed LV wall and blood pool. The proposed approach is also noted to be effective on correcting the spill-over effect in the myocardium from nearby bowel or liver activities.
    • 4D non-local means post-filtering for cardiac gated SPECT

      Song, Chao; Yang, Yongyi; Pretorius, P. Hendrik; King, Michael A. (2017-12-28)
      Cardiac gated images often suffer from increased noise in single photon emission computed tomography (SPECT) due to reduced data counts compared to non-gated studies. We investigate a spatiotemporal post-processing approach based on a non-local means (NLM) filter for suppressing the noise in gated SPECT images. In this filter, the output at a voxel location is computed from a weighted average of voxels in its 4D neighborhood, wherein the filter coefficients are adjusted according to the similarity level in the local image pattern of individual voxels with respect to the output voxel. This adaptive property allows the filter to achieve noise reduction while avoiding excessive blur of the heart wall. In the experiments, we first evaluated the accuracy of the proposed NLM filtering approach using simulated SPECT imaging data. We then demonstrated the approach on eight sets of clinical acquisitions. In addition, we also explored the robustness of the NLM filter with imaging dose reduced by 50% in these clinical acquisitions. The quantitative results show that 4D NLM filtering could effectively reduce the noise level in gated images, leading to more accurate reconstruction of the myocardium. Compared to spatial filtering alone, using temporal filtering in NLM could reduce the mean-squared-error of the myocardium by 55.63% and improve the LV resolution by 19.92%. It could also improve the visibility of perfusion defects in gated images. Similar results are also observed on the clinical acquisitions both at standard dose and at 50% reduced dose. The 4D NLM results are also found to be comparable to that of a motion-compensated 4D reconstruction approach which is computationally more demanding.
    • 4D reconstruction for low-dose cardiac gated SPECT

      Jin, Mingwu; Niu, Xiaofeng; Qi, Wenyuan; Yang, Yongyi; Dey, Joyoni; King, Michael A.; Dahlberg, Seth T.; Wernick, Miles N. (2013-02-01)
      PURPOSE: Due to the combination of high-frequency use and relatively high diagnostic radiation dose (>9 mSv for one scan), there is a need to lower the radiation dose used in myocardial perfusion imaging (MPI) studies in cardiac gated single photon emission computed tomography (GSPECT) in order to reduce its population based cancer risk. The aim of this study is to assess quantitatively the potential utility of advanced 4D reconstruction for GSPECT for significantly lowered imaging dose. METHODS: For quantitative evaluation, Monte Carlo simulation with the 4D NURBS-based cardiac-torso (NCAT) phantom is used for GSPECT imaging at half and quarter count levels in the projections emulating lower injected activity (dose) levels. Both 4D and 3D reconstruction methods are applied at these lowered dose levels, and compared with standard clinical spatiotemporal reconstruction (ST121) at full dose using a number of metrics on the reconstructed images: (1) overall reconstruction accuracy of the myocardium, (2) regional bias-variance analysis of the left ventricle (LV) wall, (3) uniformity of the LV wall, (4) accuracy of the time activity curve (TAC) of the LV wall, and (5) detectability of perfusion defects using channelized Hotelling observer. As a preliminary demonstration, two sets of patient data acquired in list-mode are used to illustrate the conservation of both image quality and LV ejection fraction (LVEF) by 4D reconstruction where only a portion of the acquired counts at each projection angle are used to mimic low-dose acquisitions. RESULTS: Compared to ST121 at standard dose, even at quarter dose 4D achieved better performance on overall reconstruction accuracy of the myocardium (28.7% improvement on relative root mean square error: standard vs 4D quarter p-value < 10(-30)), regional bias-variance analysis, and similar performance on accuracy of the TAC of the LV wall and detectability of perfusion defects. A slight degradation in uniformity of the LV wall was observed in 4D at quarter dose due to use of scatter correction which increases reconstruction variance. The reconstructed images from simulated and patient data show that 4D at quarter dose is visually comparable to ST121 at standard dose, if not better. Compared to ST121 and 3D, 4D images exhibit less noise artifacts and better definition of the LV wall. The 4D images are also observed to be more consistent between half dose and quarter dose. 4D also yields more consistent LVEF results at different count levels on the patient data. CONCLUSIONS: With various quantitative metrics 4D reconstruction is demonstrated to achieve better or comparable performance at quarter dose ( approximately 2.25 mSv, 75% dose reduction) compared to conventional clinical reconstruction at standard dose ( approximately 9 mSv). Preliminary results from two patient datasets also show that 4D at an equivalent quarter dose can achieve better performance than clinical and 3D methods at higher dose levels. Such a significant dose reduction (75%) has not been demonstrated quantitatively in previous studies in GSPECT. These promising results warrant further investigations on the lower bound of dose reduction with different reconstruction strategies and more comprehensive clinical studies with greater patient variability.
    • 4D Reconstruction with Respiratory Correction for Gated Myocardial Perfusion SPECT

      Qi, Wenyuan; Yang, Yongyi; Song, Chao; Wernick, Miles; Pretorius, P. Hendrik; King, Michael A. (2017-08-01)
      Cardiac SPECT images are known to suffer from both cardiac and respiratory motion blur. In this work, we investigate a 4D reconstruction approach to suppress the effect of respiratory motion in gated cardiac SPECT imaging. In this approach, the sequence of cardiac gated images is reconstructed with respect to a reference respiratory amplitude bin in the respiratory cycle. To combat the challenge of inherent high imaging noise, we utilize the data counts acquired during the entire respiratory cycle by making use of a motion-compensated scheme, in which both cardiac motion and respiratory motion are taken into account. In our evaluation study, we first use Monte Carlo simulated imaging data wherein the ground truth is known for quantitative comparison. We then demonstrate the proposed approach on eight sets of clinical acquisitions, in which the subjects exhibit different degrees of respiratory motion blur. The quantitative evaluation results show that 4D reconstruction with respiratory correction could effectively reduce the effect of motion blur and lead to a more accurate reconstruction of the myocardium. The mean-squared-error of the myocardium is reduced by 22%, and the LV resolution is improved by 21%. Such improvement is also demonstrated with the clinical acquisitions, where the motion blur is markedly improved in the reconstructed LV wall and blood pool. The proposed approach is also noted to be effective on correcting the spill-over effect in the myocardium from nearby bowel or liver activities.
    • 90Y labeled phosphorodiamidate morpholino oligomer for pretargeting radiotherapy

      Liu, Guozheng; Dou, Shuping; Liu, Yuxia; Wang, Yuzhen; Rusckowski, Mary; Hnatowich, Donald J. (2011-12-21)
      While (188)Re has been used successfully in mice for tumor radiotherapy by MORF/cMORF pretargeting, previous radiolabeling of the amine-derivatized cMORF with (90)Y, a longer physical half-life nuclide, was not very successful. After developing a method involving a prepurification heating step during conjugation that increases labeling efficiency and label stability, the biodistribution of (90)Y-DOTA-Bn-SCN-cMORF ((90)Y-DOTA-cMORF) was measured in normal mice and in MORF-CC49 pretargeted mice that bear LS174T tumors. Absorbed radiation doses were then estimated and compared to those estimated for (188)Re. The pharmacokinetics of the (90)Y-DOTA-cMORF in normal mice and in the pretargeted nude mice was similar to that observed previously with (99m)Tc- and (188)Re-MAG(3)-cMORFs. While the (90)Y-DOTA-cMORF cleared rapidly from normal tissues, tumor clearance was very slow and tumor radioactivity accumulation was constant for at least 7 days such that the tumor/blood (T/B) ratio increased linearly from 6 to 25 over this period. Therefore, by extrapolation, normal tissue toxicities following administration of therapeutic doses of (90)Y may be comparable to that observed for (188)Re in which the T/B increased from 5 to 20. In conclusion, radiolabeling of DOTA-cMORF with (90)Y was improved by introducing a prepurification heating step during conjugation. The (90)Y-DOTA-cMORF provided a similar T/B ratio and biodistribution to that of (188)Re-MAG(3)-cMORF and was retained well in the tumor pretargeted with MORF-CC49. Because of the longer physical half-life, the T/NT absorbed radiation dose ratios were improved in most organs and especially in blood.
    • (99m)Tc-MORF oligomers specific for bacterial ribosomal RNA as potential specific infection imaging agents

      Chen, Ling; Wang, Yi; Cheng, Dengfeng; Liu, Xinrong; Dou, Shuping; Liu, Guozheng; Hnatowich, Donald J.; Rusckowski, Mary (2013-11-01)
      PURPOSE: Radiolabeled oligomers complementary to the 16S rRNA in bacteria were investigated as bacterial infection imaging agents. METHODS AND RESULTS: Identical sequences with backbones phosphorodiamidate morpholino (MORF), peptide nucleic acid (PNA), and phosphorothioate DNA (PS-DNA) were (99m)Tc-labeled and evaluated for binding to bacterial RNA. MORF binding to RNA from Escherichia coli strains SM101 and K12 was 4- and 150-fold higher compared to PNA and PS-DNA, respectively. Subsequently MORF oligomer in fluorescence in situ hybridization showed a stronger signal with study MORF compared to control in fixed preparations of two E. coli strains and Klebsiella pneumoniae. Flow cytometry analysis showed study MORF accumulation to be 8- and 80-fold higher compared to the control in live K. pneumoniae and Staphylococcus aureus, respectively. Further, fluorescence microscopy showed increased accumulation of study MORF over control in live E. coli and K. pneumonia. Binding of (99m)Tc-study MORF to RNA from E. coli SM101 and K12 was 30.4 and 117.8pmol, respectively, per 10(10) cells. Mice with K. pneumoniae live or heat-killed (sterile inflammation) in one thigh at 90min for both (99m)Tc-study MORF and control showed higher accumulation in target thighs than in blood and all other organs expect for kidneys and small intestine. Accumulation of (99m)Tc-study MORF was significantly higher (p = 0.009) than that of the control in the thigh with sterile inflammation. CONCLUSION: A (99m)Tc-MORF oligomer complimentary to the bacterial 16S rRNA demonstrated binding to bacterial RNA in vitro with specific accumulation into live bacteria. Radiolabeled MORF oligomers antisense to the bacterial rRNA may be useful to image bacterial infection.
    • A brief evaluation of tumor imaging in mice with 99mTc-glucarate including a comparison with 18F-FDG

      Cheng, Dengfeng; Rusckowski, Mary; Wang, Yuzhen; Liu, Yuxia; Liu, Guozheng; Liu, Xinrong; Hnatowich, Donald J. (2011-01-01)
      OBJECTIVE: Recently 99mTc-glucarate, a radiolabeled glucose analogue, has been considered as a SPECT alternative to 18F-FDG and PET for non-invasive detection of certain tumors. Thus far there have been few studies on (99m) Tcglucarate for tumor imaging and fewer, if any, studies comparing (99m)Tc-glucarate with 18F-FDG. As a preliminary indication of the properties of (99m)Tc-glucarate as a possible substitute for 18F-FDG in animal studies, we have imaged mice bearing xenografts of four tumor types with (99m)Tc-glucarate and have compared in two mice with one of these tumor types the 99mTc and 18F biodistributions. METHODS: Two mice bearing SUM190 breast cancer xenografts received 1 mCi of (99m)Tc-glucarate and were imaged on a NanoSPECT/CT small animal camera. One day later, the same animals received 1 mCi of 18F-FDG and were imaged on a MosaicHP PET small animal camera. In addition, 0.5-1 mCi of (99m)Tc-glucarate only was administered to mice bearing xenografts induced by BxPC3 pancreatic cancer cells, HEK-293 renal cell carcinomas cells or HCT-116 colorectal tumor cells. NanoSPECT/CT acquisitions were performed in these mice to evaluate tumor accumulations. RESULTS: In the SUM190 xenografted mice, the average tumor accumulation was 1.4 % (ID%/cm3) for (99m)Tc-glucarate and 2.1 % (ID%/cm3) for 18F-FDG. While slightly higher than (99m)Tc-glucarate, the tumor accumulation of 18F-FDG was accompanied by higher bone marrow and muscle accumulations at levels that could interfere with the tumor image depending upon location. The whole body clearance of (99m)Tc-glucarate was faster than that of 18F-FDG. Tumor accumulation of (99m)Tc-glucarate varied among tumor types but the tumors were readily visible in all images. CONCLUSION: In a direct comparison in the same two SUM190 tumored animals, SPECT images obtained with (99m)Tcglucarate compared favorably with PET images obtained with 18F-FDG. Tumor images with 99mTc-glucarate were also positive in three additional tumor mouse models. While further comparison studies are necessary, we conclude that (99m)Tcglucarate may be a more convenient and less expensive alternative to 18F-FDG for tumored mouse studies.
    • A canine model of mechanical thrombectomy in stroke

      Brooks, Olivia W.; King, Robert M.; Nossek, Erez; Marosfoi, Miklos G.; Caroff, Jildaz; Chueh, Juyu; Puri, Ajit S.; Gounis, Matthew J. (2019-05-18)
      PURPOSE: To develop a preclinical model of stroke with a large vessel occlusion treated with mechanical thrombectomy. MATERIALS AND METHODS: An ischemic stroke model was created in dogs by the introduction of an autologous clot into the middle cerebral artery (MCA). A microcatheter was navigated to the clot and a stent retriever thrombectomy was performed with the goal to achieve Thrombolysis in Cerebral Ischemia (TICI) 2b/3 reperfusion. Perfusion and diffusion MRI was acquired after clot placement and following thrombectomy to monitor the progression of restricted diffusion as well as changes in ischemia as a result of mechanical thrombectomy. Post-mortem histology was done to confirm MCA territory infarct volume. RESULTS: Initial MCA occlusion with TICI 0 flow was documented in all six hound-cross dogs entered into the study. TICI 2b/3 revascularization was achieved with one thrombectomy pass in four of six animals (67%). Intra-procedural events including clot autolysis leading to spontaneous revascularization (n=1) and unresolved vasospasm (n=1) accounted for thrombectomy failure. In one case, iatrogenic trauma during microcatheter navigation resulted in a direct arteriovenous fistula at the level of the cavernous carotid. Analysis of MRI indicated that a volume of tissue from the initial perfusion deficit was spared with reperfusion following thrombectomy, and there was also a volume of tissue that infarcted between MRI and ultimate recanalization. CONCLUSION: We describe a large animal stroke model in which mechanical thrombectomy can be performed. This model may facilitate, in a preclinical setting, optimization of complex multimodal stroke treatment paradigms for clinical translation.
    • A case of distal limb arterial tortuosity and dilation: observations and potential clinical significance

      Carter, Yasmin; Bennett, Desmond J.; Molla, Vadim; Wink, Alexandra E.; Collins, Amanda J.; Giannaris, Eustathia Lela (2021-06-01)
      Arterial tortuosity describes variation via bending of the arterial wall and has been noted in several arteries throughout the body. Tortuous blood vessels can cause nerve compression, as well as present difficulties to surgeons and radiologists. Here we present an unusual case of multi-vessel arterial tortuosity discovered in 78-year-old Hispanic male cadaver, independent of systemic pathology. The left ulnar and right tibial arteries were dissected, and using calibrated digital calipers, their external and internal diameters were measured both at the origin site and at the site of greatest dilation. Both wall thickness and the number of inflection points were also measured. Six bends were noticed in the ulnar artery and its diameter measured 8.11 mm at its widest, with a wall thickness of 0.88mm. On the lower extremity, the right tibial artery had three bends and its diameter measured 4.86 mm at its widest, with a wall thickness of 1.32 mm. This uncommon tortuosity is not only more prone to laceration during surgery, but the bending and thickening can be mistaken for tumors. Finally, fluid dynamics can be altered, resulting in an impact on blood pressure in the extremities. Thus, raising awareness is crucial to prevent both symptoms and iatrogenic complications.
    • A case of solitary kidney with duplex collecting systems and renal vascular variants in an adult male cadaver

      Salimy, Mehdi S.; Luiselli, Gabrielle A.; Yuen, Megan; Healy, Rose C.; Shah, S. G.; Giannaris, Eustathia Lela; Das, Manas; Wink, Alexandra E. (2020-08-04)
      We describe a unique solitary kidney with duplex collecting system and vascular variation observed in an 86-year-old White male formaldehyde- and phenol-fixed cadaver during routine academic dissection. The left renal fossa was empty with an intact adrenal gland, and the right renal fossa contained a fused renal mass with apparent polarity between the superior and inferior regions and two renal pelves converging into a single ureter. There were three right renal arteries supplying the renal mass; the superior and middle arteries were noted to be postcaval and the inferior artery was precaval. There were also two right renal veins draining into the inferior vena cava and following a regional distribution with the superior vein draining the inferior portion of the renal mass. Despite generally being asymptomatic, the detection of renal anatomical variants is clinically important for appropriate patient management and surgical interventions.
    • A clinical perspective on endovascular stroke treatment biomechanics

      Ospel, Johanna M.; van der Lugt, Aad; Gounis, Matthew J.; Goyal, Mayank; Majoie, Charles B. L. M. (2021-10-11)
      Acute ischemic stroke (AIS) is caused by blockage of an arterial blood vessel in the brain by a thrombus, which interrupts oxygen supply to the brain parenchyma. The goal of endovascular stroke treatment (mechanical thrombectomy) is to restore blood flow as quickly and completely as possible. There are numerous factors that influence endovascular treatment success. They can be broadly grouped into a) factors related to blood vessels, b) factors related to the thrombus, c) factors related to endovascular treatment technique and tools and d) operator-related factors. While blood vessel and tgthro thrombus-related factors are mostly non-modifiable in the acute setting, operator and technique-related factors can be modified, and extensive research is currently being done to investigate the complex interplay of all these variables, and to optimize the modifiable factors to the maximum possible extent. In this review, we will describe these factors and how they interact with each other in detail, and outline some of their practical implications. We will conclude with a short summary and outlook on future directions for optimizing endovascular treatment success.
    • A comparison in monkeys of (99m)Tc labeled to a peptide by 4 methods

      Rusckowski, Mary; Qu, Tong; Gupta, Suresh; Ley, Arthur C.; Hnatowich, Donald J. (2001-12-26)
      Although a number of different strategies for labeling peptides with (99m)Tc have been developed, only a few studies have compared the in vivo properties of (99m)Tc when attached to different chelators. Furthermore, these comparisons are usually in mice, whereas results obtained in nonhuman primates may be expected to be more relevant to the clinical situation. METHODS: We evaluated the influence of 4 common chelators on the biodistribution in monkeys of (99m)Tc-labeled HNE-2, a 6.7-kDa peptide being investigated as an inflammation/infection imaging agent. The peptide was conjugated with the N-hydroxysuccinimide ester of mercaptoacetyltriglycine (MAG3), mercaptoacetyltriserine (MAS3), hydrazinonicotinamide (HYNIC), and the cyclic anhydride of diethylenetriaminepentaacetic acid (DTPA). After radiolabeling, each peptide was administered intravenously to rhesus monkeys with a Staphylococcus aureus-induced focal inflammation/infection. RESULTS: Quantification of radioactivity accumulation by regions of interest over 3 h after administration in monkeys showed important differences among labeling methods: For example, at 3 h, kidney accumulation varied in percentage injected dose per organ (%ID per organ) from 31 %ID per organ (HYNIC) to 18 %ID per organ (MAG3), whereas liver varied from 7.8 %ID per organ (MAG3) to 2.8 %ID per organ (MAS3). Radioactivity accumulation in the lesion was independent of labeling method. These organ accumulations were compared with that obtained earlier in mice by sacrifice and dissection also at 3 h and at the same administered dosage. In the rodent, kidney levels varied from 45 %ID per organ (HYNIC) to 12 %ID per organ (MAS3) and liver levels varied from 6.5 %ID per organ (DTPA) to 2.0 %ID per organ (MAS3). CONCLUSION: In agreement with previous work from this laboratory and elsewhere, the method of radiolabeling had an important effect on the biodistribution of (99m)Tc. Furthermore, although biodistribution results in mice should be used with caution to predict biodistributions in primates, in major organs, these results in mice and monkeys were similar.
    • A complication involving a braided hook-wire localization device

      D'Orsi, Carl J.; Swanson, Richard S.; Moss, Lawrence J.; Reale, Frank R.; Wertheimer, Michael D. (1993-05-01)
      At the authors' institution, needle localization of breast lesions with a braided hook wire involves the wire being cut 1-2 cm from the point of entry before dissection, to avoid contamination of the sterile field with the nonsterile portion of wire. During dissection, the wire is brought through the skin into the area of dissection. In one patient, fragments of wire filaments were left within the breast. Braided hook wires must be cut cleanly, the cut surface should be wiped before dissection, and the surgical area should be cleansed before closure.
    • A convenient thiazole orange fluorescence assay for the evaluation of DNA duplex hybridization stability

      Liang, Min Min; Liu, Xinrong; Nakamura, Kayoko; Chen, Xiangji; Cheng, Dengfeng; Liu, Guozheng; Dou, Shuping; Wang, Yi; Rusckowski, Mary; Hnatowich, Donald J. (2009-11-01)
      OBJECTIVE: A simple and rapid method for measuring the hybridization stability of duplexes of DNAs and other oligomers in different environments is described. When added to an oligomer duplex, the thiazole orange (TO) dye intercalates and in this state is fluorescent. Therefore, when duplex dissociation occurs, the release of TO results in a detectable change in fluorescence intensity. This assay was developed primarily to screen antisense oligomer duplexes that are stable in serum and in the cytoplasm but unstable in the presence of their target messenger RNA (mRNA). METHODS: The two antisense oligomers of this investigation were both 25 mer phosphorothioate (PS) DNAs, one directed against the RIα mRNA and the other directed against the mdr1 mRNA. The former duplex was first used in the solution studies, in most cases duplexed with a 16 mer phosphodiester (PO) complementary DNA (i.e., PS-DNA25/PO-cDNA16). Both duplexes were then tested in a series of cell studies using SK-BR-3 (RIα+), KB-G2 (mdr1++), and KB-31 (mdr1+/-) cells. RESULTS: Preliminary measurements in solution showed that maximum fluorescence was achieved when more than ten TO molecules were bound to each duplex. When a 25 mer PO-DNA or PO-RNA with the base sequence of the RIα mRNA was added, the dramatic change in fluorescence intensity that followed signified dissociation of the antisense DNA from the study duplex and reassociation with the target DNA. Kinetic measurements showed that this process was completed in about 3 min. Fluorescent measurements of SK-BR-3 (RIα+) cells incubated at 37 degrees C with the anti-RIα study duplex over time showed a maximum at the point where the loss of fluorescence due to dissociation of the study duplex, probably by an antisense mechanism, began to dominate over the increasing fluorescence due to continuing cellular accumulation. A similar result was observed in the KB-G2 (mdr1+) cells incubated with the anti-mdr1 study duplex. CONCLUSIONS: When study duplexes shown to be stable in serum were incubated with their target cells, the assay successfully detected evidence of dissociation, most likely by an antisense mechanism. Thus, a TO fluorescence assay has been developed that is capable of detecting the dissociation of DNA duplexes.
    • A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system

      Alterman, Julia F.; Godinho, Bruno M. D. C.; Hassler, Matthew R.; Ferguson, Chantal M.; Echeverria, Dimas; Sapp, Ellen; Haraszti, Reka A.; Coles, Andrew H.; Conroy, Faith; Miller, Rachael; et al. (2019-08-02)
      Sustained silencing of gene expression throughout the brain using small interfering RNAs (siRNAs) has not been achieved. Here we describe an siRNA architecture, divalent siRNA (di-siRNA), that supports potent, sustained gene silencing in the central nervous system (CNS) of mice and nonhuman primates following a single injection into the cerebrospinal fluid. Di-siRNAs are composed of two fully chemically modified, phosphorothioate-containing siRNAs connected by a linker. In mice, di-siRNAs induced the potent silencing of huntingtin, the causative gene in Huntington's disease, reducing messenger RNA and protein throughout the brain. Silencing persisted for at least 6 months, with the degree of gene silencing correlating to levels of guide strand tissue accumulation. In cynomolgus macaques, a bolus injection of di-siRNA showed substantial distribution and robust silencing throughout the brain and spinal cord without detectable toxicity and with minimal off-target effects. This siRNA design may enable RNA interference-based gene silencing in the CNS for the treatment of neurological disorders.
    • A DNA-guided Argonaute Protein Functions in DNA Replication in Thermus thermophilus [preprint]

      Jolly, Samson M.; Zhang, Han; Strittmatter, Lara; Hendricks, Gregory M.; Dhabaria, Avantika; Ueberheide, Beatrix; Zamore, Phillip D. (2019-12-09)
      Argonaute proteins use nucleic acid guides to protect organisms against transposons and viruses. In the eubacterium Thermus thermophilus, the DNA-guided Argonaute TtAgo defends against transformation by DNA plasmids. Here, we report that TtAgo also participates in DNA replication. TtAgo binds small DNA guides derived from the chromosomal region where replication terminates and associates with proteins known to act in DNA replication. T. thermophilus deploys a single type II topoisomerase, gyrase. When gyrase is inhibited, T. thermophilus relies on TtAgo to complete replication of its circular genome; loss of both gyrase and TtAgo activity produces long filaments that fail to separate into individual bacteria. We propose that the primary role of TtAgo is to help T. thermophilus disentangle the catenated circular chromosomes made by DNA replication.
    • A feasible approach to evaluate the relative reactivity of NHS-ester activated group with primary amine-derivatized DNA analogue and non-derivatized impurity

      Dou, Shuping; Virostko, John; Greiner, Dale L.; Powers, Alvin C.; Liu, Guozheng (2015-04-15)
      Synthetic DNA analogues with improved stability are widely used in life science. The 3'and/or 5' equivalent terminuses are often derivatized by attaching an active group for further modification, but a certain amount of non-derivatized impurity often remains. It is important to know to what extent the impurity would influence further modification. The reaction of an NHS ester with primary amine is one of the most widely used options to modify DNA analogues. In this short communication, a 3'-(NH2-biotin)-derivatized morpholino DNA analogue (MORF) was utilized as the model derivatized DNA analogue. Inclusion of a biotin concomitant with the primary amine at the 3'-terminus allows for the use of streptavidin to discriminate between the products from the derivatized MORF and non-derivatized MORF impurity. To detect the MORF reaction with NHS ester, S-acetyl NHS-MAG3 was conjugated to the DNA analogue for labeling with (99m)Tc, a widely used nuclide in the clinic. It was found that the non-derivatized MORF also reacted with the S-acetyl NHS-MAG3. Radiolabeling of the product yielded an equally high labeling efficiency. Nevertheless, streptavidin binding indicated that under the conditions of this investigation, the non-derivatized MORF was five times less reactive than the amine-derivatized MORF.
    • A Finite Element Method to Predict Adverse Events in Intracranial Stenting Using Microstents: In Vitro Verification and Patient Specific Case Study

      Iannaccone, Francesco; De Beule, Matthieu; De Bock, Sander; Van der Bom, Imramsjah M.J.; Gounis, Matthew J.; Wakhloo, Ajay K.; Boone, Matthieu; Verhegghe, Benedict; Segers, Patrick (2015-11-30)
      Clinical studies have demonstrated the efficacy of stent supported coiling for intra-cranial aneurysm treatment. Despite encouraging outcomes, some matters are yet to be addressed. In particular closed stent designs are influenced by the delivery technique and may suffer from under-expansion, with the typical effect of "hugging" the inner curvature of the vessel which seems related to adverse events. In this study we propose a novel finite element (FE) environment to study potential failure able to reproduce the microcatheter "pull-back" delivery technique. We first verified our procedure with published in vitro data and then replicated the intervention on one patient treated with a 4.5 x 22 mm Enterprise microstent (Codman Neurovascular; Raynham MA, USA). Results showed good agreement with the in vitro test, catching both size and location of the malapposed area. A simulation of a 28 mm stent in the same geometry highlighted the impact of the delivery technique, which leads to larger area of malapposition. The patient specific simulation matched the global stent configuration and zones prone to malapposition shown on the clinical images with difference in tortuosity between actual and virtual treatment around 2.3%. We conclude that the presented FE strategy provides an accurate description of the stent mechanics and, after further in vivo validation and optimization, will be a tool to aid clinicians to anticipate the acute procedural outcome avoiding poor initial results.