Detecting trace concentrations of banned doping agents in athlete biofluids remains a significant challenge for anti-doping analysis due to the lack of sensitivity, speed, and environmentally sustainable options with traditional methods. We have developed an innovative AI-supported surface-enhanced Raman spectroscopy (SERS) methodology here that uses engineered nanostructured plasmonic substrates combined with advanced machine learning algorithms to achieve ultra-low concentrations of banned substances. This new technology will enable the rapid, selective, and sensitive detection of doping agents within complicated biological matrices e.g., sweat, saliva, and urine. Our novel SERS platform uses metallic nanostructures to amplify the Raman signal to enable extremely precise molecular characterization with little sample preparation. To facilitate enhanced accuracy and robustness, we will use intelligent computational models to classify the processed spectral data e.g., support vector machines and deep neural networks, achieving classification accuracies as high as 98%. In addition, the AI component significantly reduces the detrimental effects of differences in the spectral data, noise interference, and peak overlap and thus increases the reliability of doping detection. The ability to detect substances at low concentration levels of nanomolar and picomolar; having a high degree of linearity with an R2 value greater than or equal to 0.98; having a low degree of variability among replicates will provide high levels of analytical accuracy. Compared to current conventional chromatographic and mass spectrometric techniques, this method requires 70% to 85% less reagents, provides quicker analysis, less than 2 min for each sample and requires less energy consumption. In addition, the use of micro-scale sensors together with data-driven analyses will facilitate the development of portable diagnostic devices that can monitor anti-doping in real time and/or on a decentralized basis. The sensitivity, specificity and scalability of this methodology for the identification of trace and sub-trace levels has been demonstrated through experimental and simulated testing. In conclusion, the combination of Surface Enhanced Raman Spectroscopy (SERS) and Artificial Intelligence (AI)-based smart intelligence computing can be viewed as a revolutionary breakthrough in the area of sustainable microchemical analysis. The resulting system has the potential to provide a viable, environmentally friendly solution for issues related to sport integrity as well as for many other applications in the field of biomedicine.
The distal tibia bone quality is of paramount importance for ankle fractures, total ankle implants, ankle fusions, and osteotomy procedures. Despite this fact relatively little is known regarding the overall bone quality for this section of the tibia. Previous literature suggest that there is a statistically significant decrease in bone mineral density within the distal 5% to 10% segment of the tibia medullary canal. This segment of medullary bone is considerable in size and thus valuable for fixation constructs as it is oftentimes utilized for medial malleolar fractures, distal tibia fractures, total ankle replacements, ankle fusions, and other procedures. This study assessed bone attenuation between the distal 5% and 10% mark of the tibia in 1% slices via Hounsfield unit measurements on CT scans based on previously established correlation between Hounsfield units and bone mineral density found on DEXA scans. One hundred five distal tibia segments were assessed with an average interval in percentile slices of 3.8 mm. As expected there was a gradual decrease in bone attenuation noted with each proximal percentile segment. There exists a statistically significant difference in bone attenuation among males versus females as well as those older than 60 years versus younger than 60 years. The findings suggest fixation constructs in the tibia medullary canal may find limited benefit proximal from 7% segment in females ≥60, or 26.1 mm from tibial plafond. Fixation constructs in tibia medullary canal may find limited benefit proximal from 8% segment in males <60, or 32.3 mm from tibial plafond.
Abstract Disclosure: A.R. Pallapati: None. J. Gimenez-Roig: None. S. Rojekar: None. F. Korkmaz: None. D. Sant: None. O. Barak: None. F. Sultana: None. O. Moldavski: None. A. Gumerova: None. H.S. Kannangara: None. U. Cheliadinova: None. C.J. Rosen: None. J.N. Caminis: None. M. Meseck: None. V.E. Demambro: None. S.L. Sims: None. S. Gera: None. R. Witztum: None. S. Miyashita: None. V. Ryu: None. M. Saxena: None. T. Frolinger: None. A. Macdonald: None. S. Kim: None. G. Pevnev: None. D. Lizneva: None. T. Yuen: None. M. Zaidi: None. Pharmacological and genetic studies suggest that FSH is an actionable target for diseases affecting millions, notably osteoporosis, obesity and Alzheimer’s disease (AD). Blocking FSH action prevents bone loss (1, 2), fat accrual (3) and AD-like features in mice (4). We recently developed a first-in-class, humanized, epitope-specific FSH blocking antibody that binds to a 13-amino-acid-long sequence of FSHβ—MS-Hu6—with a KD of 7.52 nM (5). We showed that MS-Hu6 binds specifically to FSHβ, without binding to LH and TSH. For efficacy studies, we have blocked FSH action using either MS-Hu6 or the parent murine antibody, Hf2, targeted to the same epitope. Using our Good Laboratory Practice platform (Code of Federal Regulations, Title 21, Part 58), we report that FSH blockade prevents obesity, osteoporosis and AD in mice. We injected 20-week-old C57BL/6 male mice on a high-fat diet with a range of doses of Hf2 or vehicle s.c. five-days-a-week for 8 weeks. Hf2 (100 µg/mouse/day) reduced the increase in fat mass by 33% starting week 3, with a 7% reduction in in body weight. In separate studies, MS-Hu6 not only caused beiging of white adipose tissue in UCP1-reporter ThermoMice (IVIS imaging), but also improved bone density and microstructure (micro-CT) by elevating bone formation (dynamic histomorphometry). The increase in bone mass and improved microstructure were replicated in Cliff Rosen’s lab using C57BL6 mice 24 weeks post-ovariectomy. Novel Object Recognition testing of AD-prone, ovariectomized 3xTg mice showed a deficit in recognition memory, which was reversed after 8 weeks of Hf2 (100 µg/mouse/day for 5-days-a-week) exposure. Biodistribution studies using 89Zr-labelled, biotinylated or unconjugated MS-Hu6 in mice and monkeys showed localization to bone, bone marrow, fat depots and brain tissue. MS-Hu6 displayed a β phase t½ of 7.5 days in humanized Tg32 mice. In monkeys, an acute single injection of MS-Hu6 did not affect vitals, and biochemical parameters remained within the normative range. We tested 215 variations of excipients using a range of physicochemical techniques, including protein thermal shift, size exclusion chromatography, dynamic light scattering, Fourier-transform infrared spectroscopy, circular dichroism spectroscopy, and differential scanning calorimetry, to yield a formulation with thermal, colloidal, monomeric and structural stability at an ultra-high concentration (100 mg/mL) with acceptable viscosity, clarity and turbidity parameters. MS-Hu6 showed the same “humanness” as human IgG1 in silico and was non-immunogenic in ELISPOT assays for IL-2 and IFNγ in human PBMC cultures. In conclusion, MS-Hu6 is efficacious, durable and manufacturable, and is therefore poised for human testing as a multipurpose therapeutic for obesity, osteoporosis, and perhaps for AD. 1Cell, 2006; 2PNAS, 2018,; 3Nature, 2017; 4Nature, 2022; 5. PNAS, 2020, eLife, 2022. Presentation: Saturday, June 17, 2023
Abstract Background Patients with Crohn’s disease (CD) experience intestinal inflammation. Ontamalimab (SHP647), a fully human immunoglobulin G2 monoclonal antibody against mucosal addressin cell adhesion molecule-1, is a potential novel CD treatment. OPERA II, a multicenter, open-label, phase 2 extension study, assessed the long-term safety and efficacy of ontamalimab in patients with moderate-to-severe CD. Methods Patients had completed 12 weeks of blinded treatment (placebo or ontamalimab at 22.5, 75, or 225 mg subcutaneously) in OPERA (NCT01276509) or had a clinical response to ontamalimab 225 mg in TOSCA (NCT01387594). Participants received ontamalimab at 75 mg every 4 weeks (weeks 0–72), then were followed up every 4 weeks for 24 weeks. One-time dose reduction to 22.5 mg or escalation to 225 mg was permitted at the investigator’s discretion. The primary end points were safety and tolerability outcomes. Secondary end points included changes in serum drug and biomarker concentrations. Efficacy end points were exploratory, and used non-responder imputation methods. Results Overall, 149/268 patients completed the study. The most common adverse event leading to study discontinuation was CD flare (19.8%). Two patients died; neither death was considered to be drug related. No dose reductions occurred; 157 patients had their dose escalated. Inflammatory biomarker concentrations decreased. Serum ontamalimab levels were consistent with known pharmacokinetics. Remission rates (Harvey-Bradshaw Index [HBI] ≤ 5; baseline, 48.1%; week 72, 37.3%) and response rates (baseline [decrease in Crohn’s Disease Activity Index ≥ 70 points], 63.1%; week 72 [decrease in HBI ≥ 3], 42.5%) decreased gradually. Conclusions Ontamalimab was well tolerated; treatment responses appeared to be sustained over 72 weeks. ClinicalTrials.gov ID: NCT01298492.
Goals: This US-based, retrospective claims study aimed to investigate disease burden and treatment patterns in patients with eosinophilic esophagitis (EoE), and to compare health care resource use (HCRU) in patients with EoE and matched controls without EoE. Materials and Methods: Patients with a diagnosis of EoE and >= 12 months of prediagnosis data were identified from the Truven Health MarketScan Research databases (January 2008 to September 2016) and followed up from the diagnosis date until termination of eligibility for a health plan. Patient clinical characteristics and HCRU were recorded in the 12 months before diagnosis; HCRU and treatment patterns were recorded during follow-up. HCRU in patients with EoE and matched controls was compared during the 12-month postdiagnosis period. Results: Among the 23,003 patients with EoE (mean age: 34.3 y; 64.8% male), gastroesophageal reflux disease was the most common prediagnosis condition (34.6%). After diagnosis, the most common off-label, first-line treatments were proton pump inhibitor monotherapy (52.8%) and topical corticosteroid monotherapy (21.5%). Overall, 3336 patients (14.5%) received at least 3 lines of off-label pharmacotherapy. Outpatient visits (recorded in 99.9% of patients on and postdiagnosis) were most frequently to gastroenterologists/pediatric gastroenterologists (49.5% prediagnosis, 72.6% on and postdiagnosis). Inpatient admissions and outpatient and emergency room visits were more likely in patients with EoE than in matched controls (P<0.0001). Conclusions: Patients with EoE in the USA experience a high disease burden both before and after diagnosis, which requires significant HCRU. Our findings highlight the unmet need for adequate control of EoE-related symptoms.
