Tetrazine-Derived Near-Infrared Dye as a Facile Reagent for Developing Targeted Photoacoustic Imaging Agents.

A new photoacoustic (PA) dye was developed as a simple-to-use reagent for creating targeted PA imaging agents. The lead molecule was prepared via an efficient two-step synthesis from an inexpensive commercially available starting material. With the dye's innate albumin-binding properties, the resulting tetrazine-derived dye is capable of localizing to tumor and exhibits a biological half-life of a few hours, allowing for an optimized distribution profile. The presence of tetrazine in turn makes it possible to link the albumin-binding optoacoustic signaling agent to a wide range of targeting molecules. To demonstrate the utility and ease of use of the platform, a novel PA probe for imaging calcium accretion was generated using a single-step bioorthogonal coupling reaction where high-resolution PA images of the knee joint in mice were obtained as early as 1 h post injection. Whole-body distribution was subsequently determined by labeling the probe with 99mTc and performing tissue counting following necropsy. These studies, along with tumor imaging and in vitro albumin binding studies, revealed that the core PA contrast agent can be imaged in vivo and can be easily linked to targeting molecules for organ-specific uptake.

Lacticaseibacillus rhamnosus HA-114 and Bacillus subtilis R0179 Prolong Lifespan and Mitigate Amyloid-ß Toxicity in C. elegans via Distinct Mechanisms.

Background: Recent advances linking gut dysbiosis with neurocognitive disorders such as Alzheimer's disease (AD) suggest that the microbiota-gut-brain axis could be targeted for AD prevention, management, or treatment. Objective: We sought to identify probiotics that can delay Aß-induced paralysis. Methods: Using C. elegans expressing human amyloid-ß (Aß)1-42 in body wall muscles (GMC101), we assessed the effects of several probiotic strains on paralysis. Results: We found that Lacticaseibacillus rhamnosus HA-114 and Bacillus subtilis R0179, but not their supernatants or heat-treated forms, delayed paralysis and prolonged lifespan without affecting the levels of amyloid-ß aggregates. To uncover the mechanism involved, we explored the role of two known pathways involved in neurogenerative diseases, namely mitophagy, via deletion of the mitophagy factor PINK-1, and fatty acid desaturation, via deletion of the Δ9 desaturase FAT-5. Pink-1 deletion in GMC101 worms did not modify the life-prolonging and anti-paralysis effects of HA-114 but reduced the protective effect of R0179 against paralysis without affecting its life-prolonging effect. Upon fat5 deletion in GMC101 worms, the monounsaturated C14:1 and C16:1 FAs conserved their beneficial effect while the saturated C14:0 and C16:0 FAs did not. The beneficial effects of R0179 on both lifespan and paralysis remained unaffected by fat-5 deletion, while the beneficial effect of HA-114 on paralysis and lifespan was significantly reduced. Conclusions: Collectively with clinical and preclinical evidence in other models, our results suggest that HA-114 or R0179 could be studied as potential therapeutical adjuncts in neurodegenerative diseases such as AD.

Discovery of Hishimonus diffractus Dai, Fletcher amp; Zhang, 2013 (Hemiptera: Auchenorrhyncha: Cicadellidae) in Europe, with remarks on previously recorded species of the genus.

During recent surveys on leafhoppers in urban and agroecosystem areas in Europe the Oriental Hishimonus diffractus Dai, Fletcher Zhang, 2013 (Hemiptera, Cicadellidae) has been found. This species, described from China and Thailand, is recorded here for the first time from Europe in the following countries: France, Portugal (Madeira), Spain, Gibraltar, United Kingdom, and Italy (Sicily). H. diffractus has been found associated with Jasminum and Olea (Oleaceae), Citrus (Rutaceae), Pittosporum (Pittosporaceae), Rubus (Rosaceae), Nerium (Apocynaceae) and Vitis (Vitaceae). Presence of the adults, morphological features, and some biological notes are provided to H. diffractus. Furthermore, Hishimonus hamatus Kuoh, 1976 is here reported for the first time in Spain and France. All Hishimonus species introduced in Europe are considered, and their economic importance discussed. Details on the first record and country in Europe, origin area, current distribution and host plants are given for each species.

Frequency-domain differential photoacoustic radar: theory and validation for ultrasensitive atherosclerotic plaque imaging.

Lipid composition of atherosclerotic plaques is considered to be highly related to plaque vulnerability. Therefore, a specific diagnostic or imaging modality that can sensitively evaluate plaques' necrotic core is desirable in atherosclerosis imaging. In this regard, intravascular photoacoustic (IVPA) imaging is an emerging plaque detection technique that provides lipid-specific chemical information from an arterial wall with great optical contrast and long acoustic penetration depth. While, in the near-infrared window, a 1210-nm optical source is usually chosen for IVPA applications since lipids exhibit a strong absorption peak at that wavelength, the sensitivity problem arises in the conventional single-ended systems as other arterial tissues also show some degree of absorption near that spectral region, thereby generating undesirably interfering photoacoustic (PA) signals. A theory of the high-frequency frequency-domain differential photoacoustic radar (DPAR) modality is introduced as a unique detection technique for accurate and molecularly specific evaluation of vulnerable plaques. By assuming two low-power continuous-wave optical sources at ∼1210 and ∼970 nm in a differential manner, DPAR theory and the corresponding simulation/experiment studies suggest an imaging modality that is only sensitive and specific to the spectroscopically defined imaging target, cholesterol.

Interference-free Detection of Lipid-laden Atherosclerotic Plaques by 3D Co-registration of Frequency-Domain Differential Photoacoustic and Ultrasound Radar Imaging.

As lipid composition of atherosclerotic plaques is considered to be one of the primary indicators for plaque vulnerability, a diagnostic modality that can sensitively evaluate their necrotic core is highly desirable in atherosclerosis imaging. In this regard, intravascular photoacoustic (IVPA) imaging is an emerging plaque detection modality that provides lipid-specific chemical information of arterial walls. Within the near-infrared window, a 1210-nm optical source is usually chosen for IVPA applications because lipid exhibits a strong absorption peak at that wavelength. However, other arterial tissues also show some degree of absorption near 1210 nm and generate undesirable interfering PA signals. In this study, a novel wavelength-modulated Intravascular Differential Photoacoustic Radar (IV-DPAR) modality was introduced as an interference-free detection technique for a more accurate and reliable diagnosis of plaque progression. By using two low-power continuous-wave laser diodes in a differential manner, IV-DPAR could efficiently suppress undesirable absorptions and system noise, while dramatically improving system sensitivity and specificity to cholesterol, the primary ingredient of plaque necrotic core. When co-registered with intravascular ultrasound imaging, IV-DPAR could sensitively locate and characterize the lipid contents of plaques in human atherosclerotic arteries, regardless of their size and depth.

Ultrasound basics.

Imaging technologies for in vivo functional and molecular imaging in small animals have undergone a very fast development in the last years with very intense competition to further develop resolution and molecular sensitivity. Among the imaging technologies available, ultrasound-based molecular imaging methods are of particular interest, since the use of ultrasound contrast agents allows specific and sensitive depiction of molecular targets. Together with new developments in quantification methods of targeted microbubbles, sonography represents a dynamic and seminal tool for molecular imaging.

Ultrasound contrast agents for molecular imaging.

The successful use of targeted ultrasound contrast agents (USCAs) for qualitative US-based imaging has been shown by several academic and industrial research groups in different animal models. Furthermore, techniques have been developed that enable the in-vivo quantification of targeted microbubbles (MBs). USCAs for quantitative functional and molecular imaging in small animals can be used for a more detailed characterization of new and established disease models and provide quantitative biological insights into the interaction between drug and target or target and disease in living animals. The advantages of such contrast agents in research and development are seen to be as follows: new functional or molecular findings in the complex biology of disease development, these findings can lead to new therapeutic strategies or drug candidates, a better understanding of the treatment effects of new and existing drug candidates, a more sensitive and specific characterization of early treatment effects in living animals, identification of in-vivo biomarkers for translational medicine. Further outcomes are seen in speeding up the evaluation of new drug compounds and in a reduction of the number of animals used for biomedical research.

A novel method for the measurement of proximal fibrous cap puncture force in chronic total occlusions: the effect of increasing age.

AIMS: The composition of CTO lesions changes significantly as they age. These changes may adversely affect PCI outcome. We present a robust ex vivo technique to measure the force required to puncture the proximal cap of CTOs, and to determine whether the puncture force differs according occlusion age to examine the effects of compositional changes over time on lesion stiffness. METHODS AND RESULTS: Occlusions were created in 44 rabbit femoral arteries by thrombin injection. Between two and 15 weeks following induction, vessels were harvested and tested in a custom setup to determine the force required to puncture the proximal cap. The puncture force mean values at 2, 6, 12, and 15 weeks were 0.61N, 0.78N, 1.21N and 1.52N, respectively. The puncture force required in occlusions of ≤6 weeks was significantly lower those≥12 weeks (0.72±0.10N versus 1.45±0.13N, p<0.01). Using a cutoff point of 1N, 86% of lesions≤6 weeks of age required<1N compared to 30% of those≥12 weeks. CONCLUSIONS: We have shown an objective and reproducible testing system for measuring CTO puncture force. Puncture force is correlated with occlusion age. This technique would be useful to evaluate therapies that alter CTO composition/compliance, as well as guidewire testing.

Validation of a 40 MHz B-scan ultrasound biomicroscope for the evaluation of osteoarthritis lesions in an animal model.

OBJECTIVE: To evaluate high frequency (40 MHz) B-mode ultrasound for the detection of osteoarthritis (OA) lesions of varying severity in an animal model of OA. DESIGN: Ultrasound biomicroscopy (UBM) was performed on the femoral articular surface of adult rabbits with unilateral transection of the anterior cruciate ligament at 4, 8 and 12 weeks post-surgery and on control rabbits. The articular cartilage was examined and graded macroscopically and histologically for OA lesions. Histological examination was used as a reference to determine sensitivity and specificity of ultrasonographic and macroscopic examination regarding fibrillation and ulceration of articular cartilage. RESULTS: Identification of slight surface irregularities was made possible with UBM. The sensitivity and specificity of UBM were 92.3% and 96.4%, respectively, to detect histological fibrillation and 90.9% and 97.6%, respectively, to identify histological ulceration. Macroscopic examination using India Ink had a sensitivity and specificity of 80% and 96.4%, respectively, for fibrillation and 90.9% and 90.5%, respectively, for ulceration when compared to histology. A high correlation (rsp=0.90) was found between ultrasonographic and histological scores. CONCLUSIONS: UBM of articular cartilage reflects histological structure and can accurately detect early changes such as fibrillation. UBM has the potential to be a valuable tool for the in vivo identification of early lesions of OA and for monitoring the disease or efficacy of novel therapy if it can be packaged in a minimally invasive format suitable for intra-articular imaging.