Sonothrombolysis Before and After Percutaneous Coronary Intervention Provides the Largest Myocardial Salvage in ST Segment Elevation Myocardial Infarction.

BACKGROUND: Sonothrombolysis is a therapeutic application of ultrasound with ultrasound contrast for patients with ST elevation myocardial infarction (STEMI). Recent trials demonstrated that sonothrombolysis, delivered before and after primary percutaneous coronary intervention (pPCI), increases infarct vessel patency, improves microvascular flow, reduces infarct size, and improves ejection fraction. However, it is unclear whether pre-pPCI sonothrombolysis is essential for therapeutic benefit. We designed a parallel 3-arm sham-controlled randomized controlled trial to address this. METHODS: Patients presenting with first STEMI undergoing pPCI within 6 hours of symptom onset were randomized 1:1:1 into 3 arms: sonothrombolysis pre-/post-pPCI (group 1), sham pre- sonothrombolysis post-pPCI (group 2), and sham pre-/post-pPCI (group 3). Our primary end point was infarct size (percentage of left ventricular mass) assessed by cardiac magnetic resonance imaging at day 4 ± 2. Secondary end points included myocardial salvage index (MSI) and echocardiographic parameters at day 4 ± 2 and 6 months. RESULTS: Our trial was ceased early due to the COVID pandemic. From 122 patients screened between September 2020 and June 2021, 51 patients (age 60, male 82%) were included postrandomization. Median sonothrombolysis took 5 minutes pre-pPCI and 15 minutes post-, without significant door-to-balloon delay. There was a trend toward reduction in median infarct size between group 1 (8% [interquartile range, 4,11]), group 2 (11% [7, 19]), or group 3 (15% [9, 22]). Similarly there was a trend toward improved MSI in group 1 (79% [64, 85]) compared to groups 2 (51% [45, 70]) and 3 (48% [37, 73]) No major adverse cardiac events occurred during hospitalization. CONCLUSIONS: Pre-pPCI sonothrombolysis may be key to improving MSI in STEMI. Multicenter trials and health economic analyses are required before clinical translation.

Effects of molecule hydrophobicity and structural flexibility of appended bispecific antibody on Protein A chromatography.

Appended bispecific antibody (aBsAb) with two single chain variable fragments (scFv) linked at the c-terminus of its heavy chains is one of the promising formats in bispecific therapeutics. The presence of hydrophobic and flexible scFv fragments render aBsAb molecules higher molecule hydrophobicity and structural flexibility compared to monoclonal antibody (mAb), thus making its purification more challenging. We set out to investigate how the unique molecular properties of aBsAb affect its performance on Protein A chromatography. We showed that aBsAb has a high propensity for chromatography-induced aggregation due to its high molecule hydrophobicity, and this couldn't be improved by the addition of common chaotropic salts. Moreover, the presence of chaotropic salts, such as arginine hydrochloride (Arg-HCl), retarded aBsAb elution during Protein A chromatography rather than facilitating which was widely observed in mAb Protein A elution. Nevertheless, we were able to overcome the aggregation issue by optimizing elution condition and improved aBsAb purity from 29 % to 93 % in Protein A eluate with a high molecular weight (HMW) species of less than 5 %. We also showed that the high molecular flexibility of aBsAb leads to different hydrodynamic sizes of the aBsAb molecule post Protein A elution, neutralization, and re-acidification, which are pH dependent. This is different from mAbs where their sizes do not change post neutralization even with re-exposure to acid. The above unique observations of aBsAb in Protein A chromatography were clearly explained from the perspectives of its high molecular hydrophobicity and structural flexibility.

Restoring microvascular circulation with diagnostic ultrasound and contrast agent: rationale and design of the REDUCE trial.

OBJECTIVES: This study aims to evaluate the efficacy and cost-effectiveness of sonothrombolysis delivered pre and post primary percutaneous coronary intervention (pPCI) on infarct size assessed by cardiac MRI, in patients presenting with STEMI, when compared against sham procedure. BACKGROUND: More than a half of patients with successful pPCI have significant microvascular obstruction and residual infarction. Sonothrombolysis is a therapeutic use of ultrasound with contrast enhancement that may improve microcirculation and infarct size. The benefits and real time physiological effects of sonothrombolysis in a multicentre setting are unclear. METHODS: The REDUCE (Restoring microvascular circulation with diagnostic ultrasound and contrast agent) trial is a prospective, multicentre, patient and outcome blinded, sham-controlled trial. Patients presenting with STEMI will be randomized to one of 2 treatment arms, to receive either sonothrombolysis treatment or sham echocardiography before and after pPCI. This tailored design is based on preliminary pilot data from our centre, showing that sonothrombolysis can be safely delivered, without prolonging door to balloon time. Our primary endpoint will be infarct size assessed on day 4±2 on Cardiac Magnetic Resonance (CMR). Patients will be followed up for 6 months post pPCI to assess secondary endpoints. Sample size calculations indicate we will need 150 patients recruited in total. CONCLUSIONS: This multicentre trial will test whether sonothrombolysis delivered pre and post primary PCI can improve patient outcomes and is cost-effective, when compared with sham ultrasound delivered with primary PCI. The results from this trial may provide evidence for the utilization of sonothrombolysis as an adjunct therapy to pPCI to improve cardiovascular outcomes in STEMI. ANZ Clinical Trial Registration number: ACTRN 12620000807954.

False-Positive Metastatic Bone Disease on FDG PET/CT Due to Multilevel Tophaceous Gout of the Spine.

ABSTRACT: A 74-year-old woman was referred for 18 F-FDG PET/CT for the evaluation of incidental CT finding of expansile destruction of left L4/5 facet joint with associated soft tissue mass concerning for a metastatic deposit. The FDG PET/CT revealed variable abnormally increased FDG activity involving multiple facet joints in all regions of the spine with corresponding expansile "punched-out" lytic lesions with sclerotic rims and overhanging margins on CT, raising the possibility of inflammatory polyarthropathy, including gout, as a differential diagnosis. Dual-energy CT of lumbar spine and CT-guided biopsy and culture of the left L4/5 facet joint demonstrated the presence of urate crystal deposition with no evidence of malignancy or infection, confirming the diagnosis of multilevel tophaceous gout of the spine.

Intra-individual comparison of prostate-specific membrane antigen positron emission tomography/computed tomography versus bone scan in detecting skeletal metastasis at prostate cancer diagnosis.

OBJECTIVES: To compare the diagnostic performance and radiological staging impact of 68 Ga-prostate-specific membrane antigen positron emission tomography/computed tomography (PSMA PET/CT) compared to 99 Tc whole-body bone scan (WBBS) for the detection of skeletal metastasis in the primary staging of prostate cancer (PCa). PATIENTS AND METHODS: A prospective institutional database was retrospectively examined for patients who underwent both PSMA PET and WBBS within a 1 week interval for PCa primary staging. Lesions were categorised as 'negative', 'equivocal', or 'definite' based on nuclear medicine physician interpretation. Metastatic burden was characterised for each imaging modality according to three groups: (i) local disease (no skeletal metastases), (ii) oligometastatic disease (three or fewer skeletal metastases), or (iii) polymetastatic disease (more than three skeletal metastases). RESULTS: There were 667 patients included. The median (interquartile range) prostate-specific antigen level was 9.2 (6.2-16) ng/mL and 60% of patients were high risk according to a modified D'Amico risk classification. The overall distribution of skeletal metastasis detection changed across the two scans overall (P = 0.003), being maintained within high-risk (P = 0.030) and low-risk (P = 0.018) groups. PSMA PET/CT identified more definite skeletal metastases compared to WBBS overall (10.3% vs 7.3%), and according to risk grouping (high: 12% vs 9%, intermediate: 4% vs 1%). Upstaging was more common with PSMA PET/CT than WBBS (P = 0.001). The maximum standardised uptake value (SUVmax ) of the primary tumour was associated with upstaging of skeletal metastases on PSMA PET/CT (P = 0.025), while age was associated with upstaging on WBBS (P = 0.021). The SUVmax of the primary tumour and metastases were both higher according to extent of metastatic disease (P = 0.001 and P < 0.001, respectively). CONCLUSIONS: More skeletal metastases were detected with PSMA PET/CT than WBBS, resulting in a higher upstaging rate mostly in high-risk patients. The SUVmax of the primary tumour and metastases was associated with upstaging.

Gastric Schwannoma Demonstrated on 18 F FDG PET/CT With Pathological Correlation.

ABSTRACT: A 66-year-old man with background hypertension and type 2 diabetes presented with 1 month history of epigastric pain for investigation. Abdominal CT and MRI identified round homogeneously enhancing mass arising from the lesser curvature of the stomach. On 18 F-FDG PET/CT, the gastric mass demonstrated intense FDG uptake (SUV max , 9.6). The patient subsequently underwent partial gastrectomy, with pathological features and immunohistochemical patterns consistent with gastric schwannoma.

Comparative spatial proteomics of Plasmodium-infected erythrocytes.

Plasmodium parasites contribute to one of the highest global infectious disease burdens. To achieve this success, the parasite has evolved a range of specialized subcellular compartments to extensively remodel the host cell for its survival. The information to fully understand these compartments is likely hidden in the so far poorly characterized Plasmodium species spatial proteome. To address this question, we determined the steady-state subcellular location of more than 12,000 parasite proteins across five different species by extensive subcellular fractionation of erythrocytes infected by Plasmodium falciparum, Plasmodium knowlesi, Plasmodium yoelii, Plasmodium berghei, and Plasmodium chabaudi. This comparison of the pan-species spatial proteomes and their expression patterns indicates increasing species-specific proteins associated with the more external compartments, supporting host adaptations and post-transcriptional regulation. The spatial proteome offers comprehensive insight into the different human, simian, and rodent Plasmodium species, establishing a powerful resource for understanding species-specific host adaptation processes in the parasite.

Manufacturability and functionality assessment of different formats of T-cell engaging bispecific antibodies.

T-cell-engaging bispecific antibodies (T-bsAbs) are promising immunotherapies for cancer treatment due to their capability of redirecting T-cells toward destroying tumor cells. Numerous T-bsAb formats have been developed, each with advantages and disadvantages in terms of developability, immunogenicity, effector functions, and pharmacokinetics. Here, we systematically compared T-bsAbs produced using eight different formats, evaluating the effect of molecular design of T-bsAbs on their manufacturability and functionality. These eight T-bsAb formats were constructed using antigen-binding fragments (Fabs) and single-chain variable fragments (scFvs) of antibodies linked to the crystallizable fragment (Fc) domain of immunoglobulin G. To ensure a fair comparison of growth and production data, we used recombinase-mediated cassette exchange technology to generate the T-bsAb-producing CHO cell lines. The produced T-bsAbs were assessed for their purification profile and recovery, binding capability, and biological activities. Our findings indicated that the manufacturability of bsAbs was adversely affected with increased number of scFv building blocks, while the functionality was affected by the combination of multiple factors, including the binding affinity and avidity of targeting moieties and the flexibility and geometry of formats. These results provide valuable insights into the impact of the format design on the optimal production and function of T-bsAbs.

Comparison between PSMA PET/CT and MRI for Characterizing Hepatocellular carcinoma: A Real-World Study.

Prostate specific membrane antigen (PSMA) is expressed by hepatocellular carcinoma (HCC). PSMA PET/CT has potential as an imaging agent for the detection of HCC including early diagnosis and monitoring for recurrence following surgical resection. This study aims to compare PSMA PET to standard surveillance imaging in the detection of HCC. Patients with suspected or treated HCC were prospectively recruited from a tertiary hospital outpatient clinic. In addition to routine surveillance imaging as recommended by the multidisciplinary team, a PSMA PET/CT was performed. Imaging and clinical characteristics were compared over a follow-up period of up to 12 months. In a cohort of 19 patients with known HCC or suspected recurrent HCC, PSMA PET/CT had similar efficacy to MRI for the detection of HCC, with a sensitivity of 91% and a specificity of 70% and sensitivity of 87% and a specificity of 73% for PSMA PET/CT and MRI, respectively. PSMA PET/CT had a higher negative predictive value of 90%. In this relatively large single centre study, PSMA is shown to have promising equivalence in performance and its role should be further evaluated in multi-centre prospective trials.

Strategies to enhance immunomodulatory properties and reduce heterogeneity in mesenchymal stromal cells during ex vivo expansion.

Therapies using mesenchymal stromal cells (MSCs) to treat immune and inflammatory conditions are now at an exciting stage of development, with many MSC-based products progressing to phase II and III clinical trials. However, a major bottleneck in the clinical translation of allogeneic MSC therapies is the variable immunomodulatory properties of MSC products due to differences in their tissue source, donor heterogeneity and processes involved in manufacturing and banking. This variable functionality of MSC products likely contributes to the substantial inconsistency observed in the clinical outcomes of phase III trials of MSC therapies; several trials have failed to reach the primary efficacy endpoint. In this review, we discuss various strategies to consistently maintain or enhance the immunomodulatory potency of MSCs during ex vivo expansion, which will enable the manufacture of allogeneic MSC banks that have high potency and low variability. Biophysical and biochemical priming strategies, the use of culture additives such as heparan sulfates, and genetic modification can substantially enhance the immunomodulatory properties of MSCs during in vitro expansion. Furthermore, robust donor screening, the use of biomarkers to select for potent MSC subpopulations, and rigorous quality testing to improve the release criteria for MSC banks have the potential to reduce batch-to-batch heterogeneity and enhance the clinical efficacy of the final MSC product. Machine learning approaches to develop predictive models of individual patient response can enable personalized therapies and potentially establish correlations between in vitro potency measurements and clinical outcomes in human trials.

A multi-modality medical imaging head and neck phantom: Part 1. Design and fabrication.

A novel anthropomorphic head and neck phantom which features the emulation of blood flow and perfusion is proposed. The phantom is helpful in both education and research and contains major blood vessels and a porous silicone elastomer brain compartment with microvascular capillary flow. The porous brain compartment is fabricated by use of a novel cast-moulding-dissolution technique. The skull and vertebra are fabricated by a combination of 3D printing and cast-moulding and are tissue equivalent with CT numbers ranging from 1000 HU to 1200 HU. The elastic structure of the phantom allows ultrasound imaging in the neck region. MRI compatible pressure sensors measure the pressure in the carotic arteries and the jugular veins and pulsatile flow is created by use of a peristaltic pump. The pressure-flow dynamics are physiologically realistic and also matches well with computational simulations of porous Darcy flow. The phantom can be used to optimize and validate MRI pulse sequences and protocols for flow imaging, MR angiography, Arterial Spin Labeling (ASL) and dynamic contrast enhanced (DCE) MRI.

A multi-modality medical imaging head and neck phantom: Part 2. Medical imaging.

The head and neck phantom discussed in an accompanying paper (part 1), is imaged with MRI, X-ray CT, PET and ultrasound. MRI scans show a distinct image contrast between the brain compartment and other anatomical regions of the head. The silicone matrix that was used to create a porous brain compartment has a relatively high proton density and a spin-spin relaxation time (T2) that is long enough to provide an MRI signal. While the longitudinal magnetization was found to recover according to a mono-exponential, a bi-exponential decay was observed for the transverse relaxation with a slow T2 relaxation component corresponding to the perfusate and a fast T2 relaxation component corresponding to the silicone. The fraction of the slow T2 relaxation component increases upon perfusion. A dynamic contrast enhanced (DCE) MRI experiment is conducted in which the injection rate of the contrast agent is varied. Parametric DCE maps are created and reveal regional differences in contrast agent kinetics as a result of differences in porosity. The skull, vertebra and the brain compartment are clearly visible on X-ray CT. Dynamic PET scanning has been performed while the carotic arterial input function is monitored by use of a Geiger-Müller counter. Similar regions of perfusion are found in the PET study as in the DCE MRI study. By doping the perfusate with a lipid micelle emulsion, the phantom is applicable for carotic Doppler ultrasound demonstration and validation.

Solitary 18F-fluorodeoxyglucose avid spinous process cholangiocarcinoma metastasis proven on CT guided percutaneous needle bone biopsy.

A 65-year-old female with newly diagnosed cholangiocarcinoma was referred for a FDG PET/CT for initial staging. There was a region of moderate FDG avidity localizing to the hepatic hilum representing the primary site of malignancy. An unexpected moderately FDG avid focus was demonstrated in the spinous process of the T11 vertebra with no corresponding mass lesion seen on low dose CT and no evidence of distant metastatic disease elsewhere. A percutaneous CT guided needle bone biopsy was performed which confirmed a solitary T11 spinous process metastasis on histopathology.

Solitary 68Ga-PSMA-Avid Proximal Tibial Prostatic Adenocarcinoma Metastasis Mimicking Enthesopathy.

A 76-year-old man with previously treated prostatic adenocarcinoma (Gleason 8) was referred for a Ga-prostate-specific membrane antigen PET/CT scan due to a rising serum PSA level. An intensely PSMA-avid focus was demonstrated in the left proximal tibia with no evidence of local recurrence or metastatic disease elsewhere. This was diagnosed and managed as enthesopathy. A Tc-MDP bone scan with SPECT/CT performed 9 months later confirmed an intensely osteoblastic mixed lytic/sclerotic metastasis at the left proximal tibia.

68Ga-Prostate-Specific Membrane Antigen Solitary Base of Skull Prostate Adenocarcinoma Metastasis.

A 74-year-old man with prostate adenocarcinoma (Gleason 7) was referred for Ga-prostate-specific membrane antigen (PSMA-HBED-CC) PET/CT scan for staging. Findings confirmed the prostate malignancy and demonstrated a solitary metastasis in the left skull base, with no evidence of regional or distant metastasis elsewhere. Solitary base of the skull metastasis may be subtle and easily overlooked, highlighting the need for thorough evaluation of the region.

Rapid activation of distinct members of multigene families in Plasmodium spp.

The genomes of Plasmodium spp. encode a number of different multigene families that are thought to play a critical role for survival. However, with the exception of the P. falciparum var genes, very little is known about the biological roles of any of the other multigene families. Using the recently developed Selection Linked Integration method, we have been able to activate the expression of a single member of a multigene family of our choice in Plasmodium spp. from its endogenous promoter. We demonstrate the usefulness of this approach by activating the expression of a unique var, rifin and stevor in P. falciparum as well as yir in P. yoelii. Characterization of the selected parasites reveals differences between the different families in terms of mutual exclusive control, co-regulation, and host adaptation. Our results further support the application of the approach for the study of multigene families in Plasmodium and other organisms.

A prospective, matched comparison of ultra-low and standard-dose computed tomography for assessment of renal colic.

OBJECTIVE: To determine the diagnostic accuracy of ultra-low-dose computed tomography (ULDCT) compared with standard-dose CT (SDCT) in the evaluation of patients with clinically suspected renal colic, in addition to secondary features (hydroureteronephrosis, perinephric stranding) and additional pathological entities (renal masses). PATIENTS AND METHODS: A prospective, comparative cohort study was conducted amongst patients presenting to the emergency department with signs and symptoms suggestive of renal or ureteric colic. Patients underwent both SDCT and ULDCT. Single-blinded review of the image sets was performed independently by three board-certified radiologists. RESULTS: Among 21 patients, the effective radiation dose was lower for ULDCT [mean (SD) 1.02 (0.16) mSv] than SDCT [mean (SD) 4.97 (2.02) mSv]. Renal and/or ureteric calculi were detected in 57.1% (12/21) of patients. There were no significant differences in calculus detection and size estimation between ULDCT and SDCT. A higher concordance was observed for ureteric calculi (75%) than renal calculi (38%), mostly due to greater detection of calculi of <3 mm by SDCT. Clinically significant calculi (≥3 mm) were detected by ULDCT with high specificity (97.6%) and sensitivity (100%) compared to overall detection (specificity 91.2%, sensitivity 58.8%). ULDCT and SDCT were highly concordant for detection of secondary features, while ULDCT detected less renal cysts of <2 cm. Inter-observer agreement for the ureteric calculi detection was 93.9% for SDCT and 87.8% for ULDCT. CONCLUSION: ULDCT performed similarly to SDCT for calculus detection and size estimation with reduced radiation exposure. Based on this and other studies, ULDCT should be considered as the first-line modality for evaluation of renal colic in routine practice.

False-Positive 18F-FDG PET/CT Uptake in Unilateral Lactation.

A 31-year-old woman (7 months postpartum and lactating) with multiple sclerotic bone lesions was referred for an 18F-FDG PET/CT scan for characterization. The scan demonstrated unilateral diffuse intense FDG uptake corresponding to dense soft tissue in the right breast, likely related to secretory hyperplasia. On further questioning, it was made apparent that she had only been breastfeeding from the right breast. While the left breast also demonstrated dense soft tissue to a lesser degree, no significant FDG uptake was seen. The sclerotic bone lesions were not FDG avid, likely due to a separate non-FDG avid benign condition or bony metastases from a non-FDG avid primary malignancy. This was reinforced by the fact that subsequent investigations including serial bilateral breast ultrasound and percutaneous biopsy demonstrated no definite evidence of malignancy in the bilateral breasts. The histopathology findings of an open surgical biopsy of sclerotic lesions in the left posterior ilium were also nonspecific, favouring bone dysplasia with no evidence of malignancy.

Immunomic Identification of Malaria Antigens Associated With Protection in Mice.

Efforts to develop vaccines against malaria represent a major research target. The observations that 1) sterile protection can be obtained when the host is exposed to live parasites and 2) the immunity against blood stage parasite is principally mediated by protective antibodies suggest that a protective vaccine is feasible. However, only a small number of proteins have been investigated so far and most of the Plasmodium proteome has yet to be explored. To date, only few immunodominant antigens have emerged for testing in clinical trials but no formulation has led to substantial protection in humans. The nature of parasite molecules associated with protection remains elusive. Here, immunomic screening of mice immune sera with different protection efficiencies against the whole parasite proteome allowed us to identify a large repertoire of antigens validated by screening a library expressing antigens. The calculation of weighted scores reflecting the likelihood of protection of each antigen using five predictive criteria derived from immunomic and proteomic data sets, highlighted a priority list of protective antigens. Altogether, the approach sheds light on conserved antigens across Plasmodium that are amenable to targeting by the host immune system upon merozoite invasion and blood stage development. Most of these antigens have preliminary protection data but have not been widely considered as candidate for vaccine trials, opening new perspectives that overcome the limited choice of immunodominant, poorly protective vaccines currently being the focus of malaria vaccine researches.

68Ga-DOTATATE Uptake in Solitary Pancreatic Metastasis From Clear Cell Renal Cancer.

A 64-year-old man with a lesion of the pancreatic tail was referred for Ga-DOTATATE PET/CT imaging. His medical history included previous metastatic clear cell renal cell cancer. Ga-DOTATATE PET/CT demonstrated increased DOTATATE uptake (SUVmax 10.5) in a pancreatic tail lesion. Histopathology of the resected lesion confirmed clear renal cell cancer metastasis. This case illustrates that clear cell renal cancer metastasis can demonstrate Ga-DOTATATE accumulation.