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Stability of conjugated critical reagents supporting ligand binding assays to enable biotherapeutic drug development is a universal concern. Formulation buffer employed for long-term cold storage may be key to mitigate protein aggregation issues. We investigated biophysical and functional attributes of murine mAb and human multispecific drug labeled with biotin, ruthenium, and Alexa fluor 647 frozen at -80 °C in PBS or a protein storage buffer for 3-15 months. Aggregation was observed at 4 months in mAb A-Ru (11.2%) and -Alexa (10%) in PBS followed by precipitation and reduced biological binding at 15 months. Increased aggregation in drug Ru (11.7%, 6 months) and Alexa (6.9%, 15 months) were noted but without impact on performance. There were no observations with biotin labeled reagents., Competing Interests: Declaration of competing interest The authors report no competing or conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Coronavirus disease 2019 (COVID-19) has led to a national health care emergency in the United States and exposed resource shortages, particularly of health care providers trained to provide critical or intensive care. This article describes how digital health technologies are being or could be used for COVID-19 mitigation. It then proposes the National Emergency Tele-Critical Care Network (NETCCN), which would combine digital health technologies to address this and future crises. Methods: Subject matter experts from the Society of Critical Care Medicine and the Telemedicine and Advanced Technology Research Center examined the peer-reviewed literature and science/technology news to see what digital health technologies have already been or could be implemented to (1) support patients while limiting COVID-19 transmission, (2) increase health care providers' capability and capacity, and (3) predict/prevent future outbreaks. Results: Major technologies identified included telemedicine and mobile care (for COVID-19 as well as routine care), tiered telementoring, telecritical care, robotics, and artificial intelligence for monitoring. Several of these could be assimilated to form an interoperable scalable NETCCN. NETCCN would assist health care providers, wherever they are located, by obtaining real-time patient and supplies data and disseminating critical care expertise. NETCCN capabilities should be maintained between disasters and regularly tested to ensure continual readiness. Conclusions: COVID-19 has demonstrated the impact of a large-scale health emergency on the existing infrastructures. Short term, an approach to meeting this challenge is to adopt existing digital health technologies. Long term, developing a NETCCN may ensure that the necessary ecosystem is available to respond to future emergencies.

Introduction: This article presents an emerging capability to project damage control procedures far forward for situations where evacuation to a formal surgical team is delayed. Specifically, we demonstrate the plausibility of using a wearable augmented reality (AR) telestration device to guide a nonsurgeon through a damage control procedure., Methods: A stand-alone, low-profile, commercial-off-the-shelf wearable AR display was utilized by a remotely located surgeon to synchronously guide a nonsurgeon through proximal control of the distal external iliac artery on a surgical manikin. The manikin wound pattern was selected to simulate a rapidly exsanguinating junctional hemorrhage not controllable by nonsurgical means., Results: This capability demonstration displayed successful use of AR technology, telecommunication, and procedural training and guidance in a single test pilot. The assisted physician assistant was able to rapidly control the simulated external iliac artery injury on this model. The telestration system used was commercially available for use with available civilian cell phone, wireless and satellite networks, without the need for dedicated high-speed networks., Conclusions: A nonsurgeon, using a wearable commercial on-visual-axis telestration system, successfully performed a damage control procedure, demonstrating the plausibility of this approach., (Published by Oxford University Press on behalf of the Association of Military Surgeons of the United States 2020.)

Medical simulation training has progressed in its use of incorporating various technologies to provide quality training interfaces from novices to experts. The purpose of this paper is to explore modeling, simulation and visualization training technology interfaces to improve precision learning, rigorous, objective assessment, and performance improvement feedback for clinical procedural skill training and sustainment. Technologies to include augmented reality (AR), haptic technology and computer vision will be defined and clarified. It is believed that by exploring the combination of using AR, haptics and computer vision technologies it is possible to develop a fully immersive learning system that can automate mentoring while detecting and measuring gross and fine motor skills. Such a system can be used to predict or delay the onset of skills decay (SD) by capturing rigorous, objective measures, and human performance metrics that can provide feedback to individual performers for skills improvement in real time., (Published by Oxford University Press on behalf of the Association of Military Surgeons of the United States 2019.)

Objectives: We describe a simulation-enhanced ultrasonography (US) curriculum for first-year medical students as part of a comprehensive curricular integration of US skills. Our goal was to assess student knowledge and performance of US and determine their satisfaction with the integrated curriculum., Methods: A committee of basic science, clinical, and interinstitutional faculty developed 7 educational US modules integrated into existing anatomy and physiology courses. First-year students in years 2012 through 2014 were administered a demographic survey and a knowledge-based pretest at the outset of the US program and assessed with a posttest, satisfaction survey, and their image acquisition abilities in an objective structured clinical examination with standardized patients on completion of the program., Results: Data from 390 students showed a significant increase in knowledge from the pretest to the posttest [t (389)  = 58.027; P < .0001]. Students with higher spatial abilities or some previous US experience performed better on the posttest. The objective structured clinical examination results showed that about 83% of the students were able to capture acceptable or marginally acceptable images. Ninety-five percent of students indicated that the US educational experience enhanced their medical education., Conclusions: Initial results show that we were able to successfully develop, implement, and evaluate performance of first-year medical students on their fundamental knowledge and performance of basic US using a model that emphasized hands-on simulation-enhanced training. Furthermore, most students found the experience to be a beneficial component of their education and indicated a desire for more US training in the medical curricula., (© 2017 by the American Institute of Ultrasound in Medicine.)

Background: As it addresses both technical and nontechnical skills, simulation-based training is playing an increasingly important role in surgery. In addition to the focus on skill acquisition, it is also important to ensure that surgeons are able to perform a variety of tasks in unique and challenging situations. These situations include responding to mass casualties, dealing with disease outbreaks, and preparing for wartime missions. Simulation-based training can be a valuable training modality in these situations, as it allows opportunities to practice and prepare for high-risk and often low-frequency events., Methods: During the 8th Annual Meeting of the Consortium of the American College of Surgeons-Accredited Education Institutes in March 2015, a multidisciplinary panel was assembled to discuss how simulation can be used to prepare the surgical community for such high-risk events., Conclusion: An overview of how simulation has been used to address needs in each of these situations is presented., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Palifosfamide, the DNA-alkylating metabolite of ifosfamide (IFOS), has been synthesized as a stabilized tris or lysine salt and found to have preclinical and clinical antitumor activity. Stabilized palifosfamide overcomes limitations of IFOS because of patient-to-patient variability in response resulting from variable prodrug activation, resistance and toxicities of metabolic byproducts, acrolein and chloroacetaldehyde. Palifosfamide represents an effective alternative to IFOS and other DNA-alkylating prodrugs. The antitumor activities of stabilized palifosfamide were investigated in vivo. Dose response, route and schedule of administration, and interaction with docetaxel or doxorubicin were investigated in NCr-nu/nu mice bearing established orthotopic mammary MX-1 tumor xenografts. Oral activity was investigated in P388-1 leukemia in CD2F1 mice. Oral and intraperitoneal bioavailabilities were compared in Sprague-Dawley rats. Stabilized palifosfamide administered by optimized regimens suppressed MX-1 tumor growth (P<0.05) by greater than 80% with 17% complete antitumor responses and up to three-fold increase in time to three tumor doublings over controls. Median survival in the P388-1 (P<0.001) model was increased by 9 days over controls. Oral bioavailability in rats was 48-73% of parenteral administration, and antitumor activity in mice was equivalent by both routes. Treatment with palifosfamide-tris combined with docetaxel or doxorubicin at optimal regimens resulted in complete tumor regression in 62-75% of mice. These studies support investigation of stabilized palifosfamide in human cancers by parenteral or oral administration as a single agent and in combination with other approved drugs. The potential for clinical translation of the cooperative interaction of palifosfamide-tris with doxorubicin by intravenous administration is supported by results from a recent randomized Phase-II study in unresectable or metastatic soft-tissue sarcoma.

Collaboration is used by the US National Security Council as a means to integrate inter-federal government agencies during planning and execution of common goals towards unified, national security. The concept of collaboration has benefits in the healthcare system by building trust, sharing resources, and reducing costs. The current terrorist threats have made collaborative medical training between military and civilian agencies crucial. This review summarizes the long and rich history of collaboration between civilians and the military in various countries and provides support for the continuation and improvement of collaborative efforts. Through collaboration, advances in the treatment of injuries have been realized, deaths have been reduced, and significant strides in the betterment of the Emergency Medical System have been achieved. This review promotes collaborative medical training between military and civilian medical professionals and provides recommendations for the future based on medical collaboration.

In the cytochrome c-551 family, the heme 17-propionate caboxylate group is always hydrogen bonded to an invariant Trp-56 and conserved residues (His and Arg mainly, Lys occasionally) at position 47. The mutation of His-47 to Ala-47 for Pseudomas stutzeri ZoBell cytochrome c-551 removes this otherwise invariant hydrogen bond. The solution structure of ferrous H47A has been solved based on NMR-derived constraints. Results indicate that the mutant has very similar main chain folding compared to wild-type. However, less efficient packing of residues in the mutant surrounding the heme propionates leads to more solvent exposure for both propionate groups, which may account for decreased stability of the mutant. The mutant has a reduction potential different from wild-type, and furthermore, the pH dependence of this potential is not the same as for wild-type. The structure of the mutant suggests that these changes are related to the loss of the residue-47 propionate hydrogen bond and the loss of charge on the side chain of residue 47.

Olive fruit fly, Bactrocera oleae (Gmelin), was monitored with adult captures by season and trap type, and was related to fruit volume and nonharvested fruit to elucidate the occurrence of the newly introduced pest in California. The highest numbers of adults captured in ChamP traps in olive trees, Olea europaea, were in October in an inland valley location, and in September in a coastal location. Comparisons of trap types showed that the number of olive fruit fly adults captured in Pherocon AM traps in a commercial orchard was significantly greater than in ChamP traps. A significantly greater number of females were captured in Pherocon AM traps with bait packets and pheromone lures than traps with pheromone lures alone, while a significantly greater number of adults and males were captured in traps with pheromone lures alone. Significantly more adults were captured in ChamP traps with bait packets and pheromone lures versus traps with bait packets alone. Fruit volume increased by four times from mid-June to mid-November. Olive fruit fly was found to oviposit on small olive fruit <1 cm3 shortly after fruit set, the maximum number of ovipositional sites per fruit occurred in October, and the greatest number of pupae and adults were reared from fruit collected in September and October. The highest numbers of pupae were collected from nonharvested fruit in March when high numbers of adults were captured in the same orchard.

Objectives: To assess the individual and team skills acquired from an interactive training program to prepare emergency personnel to respond to terrorist acts., Methods: We developed a 16-hour, two-day, multimedia- and simulation-enhanced course that places learners in realistic situations using the equipment required to respond to various chemical, biologic, radiologic, and explosive acts of terrorism. Small-group sessions and drills were conducted. Errors in skill performance were corrected immediately, and then skills were repeated to achieve mastery. Participants included emergency medical technicians, paramedics, nurses, and physicians. Team performance was assessed over four successive scenarios using a 100-mm visual analog scale. Individual learner skill acquisition was assessed with precourse and postcourse evaluation of selected skills in a randomized sampling of consenting learners., Results: Nearly all teams achieved mastery of the required skills by the second assessment rotation. Individual learners demonstrated significant gains in the ability to emergently don personal protective equipment and administer a nerve agent antidote kit., Conclusions: An interactive, simulation-enhanced curriculum of terrorism response training for emergency responders can produce significant, quantifiable individual and team skill gain. Future studies should further address performance benchmarks for these newly acquired skills.

Endothelial phenotypes derived from different pulmonary vascular segments have markedly different permeability response to inflammatory agonists, but their responses to mechanical strain have not been characterized. Therefore, we evaluated the effect of cyclical stretch on cell shape, cell membrane wounding, and junctional beta-catenin in rat pulmonary artery (RPAEC) and microvascular (RPMVEC) endothelial cell monolayers. After 24 h of 24% uniaxial strain at 40 cycles/min, RPAEC but not RPMVEC reoriented transverse to the axis of strain. Total beta-catenin increased in RPAEC but decreased in RPMVEC. Transient plasma membrane wounding was produced by cyclical biaxial strain of 34% or by scratching of monolayers with a needle and was indicated by retention of lysine fixable fluorescent 70 kDa dextran. Junctional beta-catenin was quantified by fluorescence intensity and image analysis. beta-catenin fluorescence was significantly lower in wounded cells than in adjacent uninjured cells in both phenotypes, and the decrease was significantly greater in RPAEC compared to RPMVEC in both scratched (57% vs. 30%) and stretched (55% vs. 37%) cells. Using immunoprecipitation, VE-cadherin-associated beta-catenin decreased significantly in RPAEC (61%) but E-cadherin-associated beta-catenin was not significantly decreased in RPMVEC after 34% biaxial cyclical strain. These data suggest that RPAEC more readily remodel cell-cell adhesions during cyclical stretch than RPMVEC and that a reduced intercellular adhesion adjacent to wounded cells could serve as transvascular leak sites in both phenotypes.

Introduction: Responding to acts of terrorism requires the effective use of public-safety and medical-response resources. The knowledge, skills and attitudes necessary to respond to future threats is unfamiliar to most emergency responders., Objectives: The purpose of this report to describe the development, implementation and evaluation of a multidisciplinary, interactive and simulation-enhanced course to prepare responders to acts of terrorism., Methods: We used a 5-step systematic process to develop a blended-learning, simulation-enhanced training program. Learners completed a self-confidence questionnaire and written examination prior to the course and a self-confidence questionnaire, written examination and course evaluation when they finished the course., Results: From July 7, 2003 to March 8, 2005, 497 consenting learners completed the course. After course completion, learners demonstrated significant increases in their knowledge of terrorism response (t = -64.3, df = 496, p < 0.05) and their confidence in responding to terrorist events (t = -45.5, df = 496, p < 0.05). Learner feedback about the course was highly positive., Conclusions: We successfully implemented a two-day course for professionals likely to respond to terrorist acts that included scenario-based performance training and assessment. Course participants increased their knowledge and were more confident in their ability to respond to acts of terrorism after participating in this course.

The amino boronic dipeptide, PT-100 (Val-boro-Pro), a dipeptidyl peptidase (DPP) inhibitor, has been shown to up-regulate gene expression of certain cytokines in hematopoietic tissue via a high-affinity interaction, which appears to involve fibroblast activation protein. Because fibroblast activation protein is also expressed in stroma of lymphoid tissue and tumors, the effect of PT-100 on tumor growth was studied in mice in vivo. PT-100 has no direct cytotoxic effect on tumors in vitro. Oral administration of PT-100 to mice slowed growth of syngeneic tumors derived from fibrosarcoma, lymphoma, melanoma, and mastocytoma cell lines. In WEHI 164 fibrosarcoma and EL4 and A20/2J lymphoma models, PT-100 caused regression and rejection of tumors. The antitumor effect appeared to involve tumor-specific CTL and protective immunological memory. PT-100 treatment of WEHI 164-inoculated mice increased mRNA expression of cytokines and chemokines known to promote T-cell priming and chemoattraction of T cells and innate effector cells. The role of innate activity was further implicated by observation of significant, although reduced, inhibition of WEHI 164 and A20/2J tumors in immunodeficient mice. PT-100 also demonstrated ability to augment antitumor activity of rituximab and trastuzumab in xenograft models of human CD20(+) B-cell lymphoma and HER-2(+) colon carcinoma where antibody-dependent cytotoxicity can be mediated by innate effector cells responsive to the cytokines and chemokines up-regulated by PT-100. Although CD26/DPP-IV is a potential target for PT-100 in the immune system, it appeared not to be involved because antitumor activity and stimulation of cytokine and chemokine production was undiminished in CD26(-/-) mice.

A dose-response relationship was not observed in olive fruit fly, Bactrocera oleae (Gmelin), larvae exposed to acetic acid concentrations (0-2.5%) used in commercial brine solutions to cure olives. Immersion in a 1% acetic acid brine solution impeded emergence of the immature stages. A 1-wk exposure of olives infested with olive fruit fly larvae to low-temperature storage as a postharvest treatment at 0-1 degree C resulted in 8% survival of the population, and exposures of 2 through 5 wk further reduced pupal and adult emergence to <1.0%. One- to 2-wk exposures at 2-3 degrees C resulted in a significant decrease in survival from 20 to 3%, respectively, and longer durations of 3-5 wk reduced survival to <1.0%. Mean daily fruit pulp temperatures in olives in the top, middle, and bottom of plastic bins stored at 2-3 degrees C decreased by 5-8 degrees C from the first to the second day. Lowest temperatures were observed in the top, and highest temperatures were observed in the middle layer of fruit, which attained a mean temperature of 3.8 degrees C on day 5. Laboratory choice tests showed that olive fruit fly oviposited at a higher rate in late season Mission olives that were green than in fruit that were in the red blush maturity stage in tests with 1- and 3-4-d exposure periods, and an increase in duration of exposure was related to an increase in the total number of ovipositional sites. Higher percentages of olive fruit fly third instars, pupae, and adults were reared from green fruit than from fruit in the red blush stage after a 1-d exposure to oviposition. Manzanillo olives were more attractive for oviposition by olive fruit fly than Mission olives, and significantly more third instars, pupae, and adults developed in Manzanillo fruit than in Mission fruit in the red blush stage. These differences were related to the better quality and higher flesh content of the Manzanillo versus Mission olives used in the tests.

In hematopoiesis, cytokine levels modulate blood cell replacement, self-renewal of stem cells, and responses to disease. Feedback pathways regulating cytokine levels and targets for therapeutic intervention remain to be determined. Amino boronic dipeptides are orally bioavailable inhibitors of dipeptidyl peptidases. Here we show that the high-affinity inhibitor Val-boro-Pro (PT-100) can stimulate the growth of hematopoietic progenitor cells in vivo and can accelerate neutrophil and erythrocyte regeneration in mouse models of neutropenia and acute anemia. Hematopoietic stimulation by PT-100 correlated with increased cytokine levels in vivo. In vitro, PT-100 promoted the growth of primitive hematopoietic progenitor cells by increasing granulocyte-colony-stimulating factor (G-CSF), interleukin-6 (IL-6), and IL-11 production by bone marrow stromal cells. Two molecular targets of PT-100 are expressed by stromal cells- CD26/DPP-IV and the closely related fibroblast activation protein (FAP). Because PT-100 was active in the absence of CD26, FAP appears to be the hematopoietic target for PT-100. Interaction of PT-100 with the catalytic site seems to be required because amino-terminal acetylation of PT-100 abrogated enzyme inhibition and hematopoietic stimulation. PT-100 is a therapeutic candidate for the treatment of neutropenia and anemia. The data support increasing evidence that dipeptidyl peptidases can regulate complex biologic systems by the proteolysis of signaling peptides.

Hessian fly puparia (37,167), Mayetiola destructor (Say), did not survive a large-scale commercial test (three freight containers) of a new quarantine treatment using compression (32 kg/cm2) and hydrogen phosphide fumigation (61 g/28.2 m3) for large-size, polypropylene fabric-wrapped bales of hay exported to Japan. Mean +/- SEM temperatures in the large-size bales in different locations in the freight containers ranged from 18.0 +/- 0.9 to 26.0 +/- 1.3 degrees C during the 7-d test conducted in a heated building at 20.1 +/- 1.1 degrees C. Highest concentrations of hydrogen phosphide in most locations in the freight containers were observed after 3 d of fumigation and ranged from 366.7 +/- 96.1 to 425.0 +/- 162.7 ppm (mean +/- SEM) and throughout the 7 d of fumigation ranged from 253.6 +/- 59.9 to 407.1 +/- 76.5 ppm (mean +/- SEM). Hydrogen phosphide residues after fumigation and aeration were <10 ppb and were below the U.S. Environmental Protection Agency tolerance of 0.1 ppm in animal feeds. The results of the test fulfills regulatory agency testing requirements and confirms the efficacy of the treatment to control Hessian fly in large-size, polypropylene fabric-wrapped bales of hay. Hessian fly puparia (2,160) did not survive a large-scale commercial test of compression (105 kg/cm2) and a 3-d hydrogen phosphide (60 g/28.3 m3) fumigation for standard bales.

Interspecific studies indicate that sperm morphology and other ejaculatory traits diverge more rapidly than other types of character in Drosophila and other taxa. This pattern has largely been attributed to postcopulatory sexual selection involving interaction between the sexes. Such divergence has been suggested to lead rapidly to reproductive isolation among populations and thus to be an 'engine of speciation.' Here, we test two critical predictions of this hypothesis: (i) there is significant variation in reproductive traits among incipient species; and (ii) divergence in interacting sex-specific traits exhibits a coevolutionary pattern among populations within a species, by examining geographical variation in Drosophila mojavensis, a species in the early stages of speciation. Significant among-population variation was identified in sperm length and female sperm-storage organ length, and a strong pattern of correlated evolution between these interacting traits was observed. In addition, crosses among populations revealed coevolution of male and female contributions to egg size. Support for these two important predictions confirms that coevolving internal characters that mediate successful reproduction may play an important part in speciation. The next step is to determine exactly what that role is.

In Drosophila, sperm length and the length of the females' primary sperm-storage organ have rapidly coevolved through post-copulatory sexual selection. This pattern is evident even among geographic populations of Drosophila mojavensis. To understand better these traits of potential importance for speciation, we performed quantitative genetic analysis of both seminal receptacle length and sperm length in two divergent populations. Parental strains, F1, F1 reciprocal (F1r), F2, F2r, backcross and backcross reciprocal generations were used in a line-cross (generation means) analysis. Seminal receptacle length is largely an autosomal additive trait, whereas additivity, dominance and epistasis all contributed to the means of sperm length. Either an X-chromosome or a Y-chromosome effect was necessary for models of sperm length to be significant. However, the overall contributions from the X and Y chromosomes to sperm length was small.

The nirM gene encoding cytochrome c-551 from Pseudomonas stutzeri Zobell (PZ) has been expressed in Escherichia coli at levels higher than those previously reported but only under strict anaerobic growth conditions. Expression yields for wild-type cytochrome in this study typically reached 0.6 micromol per liter of saturated E. coli culture (5.5mg/L). Culture conditions investigated are compared to obtained c-551 expression levels; the results may lead to a greater understanding of the challenges encountered when expressing c-type hemoproteins in E. coli. The nirM gene was mutated to produce a histidine-47-alanine mutation of c-551 that been heterologously expressed in E. coli using optimum culture conditions and had its physiochemical properties compared to those of the wild-type protein. In PZ, the histidine-47 residue is part of a conserved hydrogen-bonding network located at the bottom of the heme crevice that also involves tryptophan-56 and a heme propionate. Ionization events within this network are experimentally demonstrated to modulate c-551 oxidation-reduction potential and its observed dependence on pH around neutrality. The redox potential of the mutant cytochrome still displays pH-dependence; however, the midpoint potential is approximately 25mV lower with respect to wild-type c-551 at neutral pH while the pK at which the heme propionate (HP-17) ionizes is lowered by 1.3 pH units. Temperature and chemical denaturant studies also show that loss of the hydrogen-bond-donating imidazole leads to a large decrease in c-551 tertiary stability.

Despite its central role in post-copulatory sexual selection, the female reproductive tract is poorly understood. Here we provide the first experimental study of the adaptive significance of variation in female sperm-storage organ morphology. Using populations of Drosophila melanogaster artificially selected for longer or shorter seminal receptacles, we identify relationships between the length of this primary sperm-storage organ and the number of sperm stored, pattern of progeny production, rate of egg fertilization, remating interval, and pattern of sperm precedence. Costs and benefits of relatively short or long organs were identified. Benefits of longer receptacles include increased sperm-storage capacity and thus progeny production from a single insemination. Results suggest that longer receptacles have not naturally evolved because of developmental time costs and a correlated reduction in longevity of mated females. This latter cost may be a consequence of sexual conflict mediated by ejaculate toxicity. Receptacle length did not alter the pattern of sperm precedence, which is consistent with data on the co-evolution of sperm and female receptacle length, and a pattern of differential male fertilization success being principally determined by the interaction between these male and female traits.