Search

There is a pressing need to increase the efficiency and reliability of toxicological safety assessment for protecting human health and the environment. While conventional toxicology tests rely on measuring apical changes in vertebrate models, there is increasing interest in the use of molecular information from animal and in vitro studies to inform safety assessment. One promising and pragmatic application of molecular information involves the derivation of transcriptomic points of departure (tPODs). Transcriptomic analyses provide a snapshot of global molecular changes that reflect cellular responses to stressors and progression toward disease. A tPOD identifies the dose level below which a concerted change in gene expression is not expected in a biological system in response to a chemical. A common approach to derive such a tPOD consists of modeling the dose-response behavior for each gene independently and then aggregating the gene-level data into a single tPOD. While different implementations of this approach are possible, as discussed in this manuscript, research strongly supports the overall idea that reference doses produced using tPODs are health protective. An advantage of this approach is that tPODs can be generated in shorter term studies (e.g., days) compared to apical endpoints from conventional tests (e.g., 90-day sub-chronic rodent tests). Moreover, research strongly supports the idea that reference doses produced using tPODs are health protective. Given the potential application of tPODs in regulatory toxicology testing, rigorous and reproducible wet and dry laboratory methodologies for their derivation are required. This review summarizes the current state of the science regarding the study design and bioinformatics workflows for tPOD derivation. We identify standards of practice and sources of variability in tPOD generation, data gaps, and areas of uncertainty. We provide recommendations for research to address barriers and promote adoption in regulatory decision making., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society of Toxicology.)

In the aftermath of the COVID-19 pandemic, opportunities to modulate biological pathways common to the lifecycles of viruses need to be carefully considered. N -linked glycosylation in humans is mediated exclusively by the oligosaccharyltransferase complex and is frequently hijacked by viruses to facilitate infection. As such, STT3A/B, the catalytic domain of the OST complex, became an intriguing drug target with broad-spectrum antiviral potential. However, due to the critical role N -linked glycosylation plays in a number of fundamental human processes, the toxicological ramifications of STT3A/B inhibition required attention commensurate to that given to antiviral efficacy. Herein, we describe how known STT3A/B inhibitor NGI-1 inspired the discovery of superior tool compounds which were evaluated in in vitro efficacy and translational safety (e.g., CNS, cardiovascular, liver) studies. The described learnings will appeal to those interested in the therapeutic utility of modulating N -linked glycosylation as well as the broader scientific community.

Cardiovascular toxicity is an important cause of drug failures in the later stages of drug development, early clinical safety assessment, and even postmarket withdrawals. Early-stage in vitro assessment of potential cardiovascular liabilities in the pharmaceutical industry involves assessment of interactions with cardiac ion channels, as well as induced pluripotent stem cell-derived cardiomyocyte-based functional assays, such as calcium flux and multielectrode-array assays. These methods are appropriate for the identification of acute functional cardiotoxicity but structural cardiotoxicity, which manifests effects after chronic exposure, is often only captured in vivo. CardioMotion is a novel, label-free, high throughput, in vitro assay and analysis pipeline which records and assesses the spontaneous beating of cardiomyocytes and identifies compounds which impact beating. This is achieved through the acquisition of brightfield images at a high framerate, combined with an optical flow-based python analysis pipeline which transforms the images into waveform data which are then parameterized. Validation of this assay with a large dataset showed that cardioactive compounds with diverse known direct functional and structural mechanisms-of-action on cardiomyocytes are identified (sensitivity = 72.9%), importantly, known structural cardiotoxins also disrupt cardiomyocyte beating (sensitivity = 86%) in this method. Furthermore, the CardioMotion method presents a high specificity of 82.5%., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Introduction of monovalent COVID-19 mRNA vaccines in late 2020 helped to mitigate disproportionate COVID-19-related morbidity and mortality in U.S. nursing homes (1); however, reduced effectiveness of monovalent vaccines during the period of Omicron variant predominance led to recommendations for booster doses with bivalent COVID-19 mRNA vaccines that include an Omicron BA.4/BA.5 spike protein component to broaden immune response and improve vaccine effectiveness against circulating Omicron variants (2). Recent studies suggest that bivalent booster doses provide substantial additional protection against SARS-CoV-2 infection and severe COVID-19-associated disease among immunocompetent adults who previously received only monovalent vaccines (3).* The immunologic response after receipt of bivalent boosters among nursing home residents, who often mount poor immunologic responses to vaccines, remains unknown. Serial testing of anti-spike protein antibody binding and neutralizing antibody titers in serum collected from 233 long-stay nursing home residents from the time of their primary vaccination series and including any subsequent booster doses, including the bivalent vaccine, was performed. The bivalent COVID-19 mRNA vaccine substantially increased anti-spike and neutralizing antibody titers against Omicron sublineages, including BA.1 and BA.4/BA.5, irrespective of previous SARS-CoV-2 infection or previous receipt of 1 or 2 booster doses. These data, in combination with evidence of low uptake of bivalent booster vaccination among residents and staff members in nursing homes (4), support the recommendation that nursing home residents and staff members receive a bivalent COVID-19 booster dose to reduce associated morbidity and mortality (2)., Competing Interests: All authors have completed and submitted the International Committee of Medical Journal Editors form for disclosure of potential conflicts of interest. Stefan Gravenstein and David H. Canaday are recipients of support from the U.S. Department of Veterans Affairs and investigator-initiated grants to their universities from the National Institute of Allergy and Infectious Diseases (NIAID) to study influenza vaccine and COVID-19 in the nursing home, Pfizer to study pneumococcal vaccines, and from Sanofi Pasteur and Seqirus to study influenza vaccines. Stefan Gravenstein also performs consulting work for Janssen, Merck, Moderna, Novavax, Pfizer, Sanofi, Seqirus, and Vaxart; has served on the speaker’s bureaus for Seqirus and Sanofi; and was paid to chair data safety monitoring boards from Longeveron and SciClone. David H. Canaday has performed consulting work for Seqirus. Elizabeth M. White reports support from the National Institute on Aging, and membership on the Society for Post-acute and Long-term Care Medicine Workforce Development Committee and on the John Hartford Foundation Moving Forward Coalition Workforce Committee. Jürgen Bosch is the cofounder and Chief Executive Officer of InterRayBio, LLC. Yi Cao, Kerri St. Denis, and Alejandro B. Balazs report support from the Ragon Institute of Massachusetts General Hospital, the Massachusetts Institute of Technology, and Harvard University for equipment used in the current study. Kevin W. McConeghy reports grant support from Sanofi-Pasteur, Sequirus Pharmaceuticals, Genentech, and Janssen, unrelated to the current work. Eleftherios Mylonakis reports institutional support from the Biomedical Advanced Research and Development Authority, NIAID, the National Institute of General Medical Sciences, National Institutes of Health, Leidos Biomedical Research, Inc., Regeneron, Pfizer, Chemic lags/KODA therapeutics, Cidara, the National Cancer Institute, and SciClone Pharmaceuticals, and receipt of consulting fees from Basilea Pharmaceutica International, Ltd. Christopher L. King reports National Cancer Institute support for Early Drivers of Humoral Immunity to SARS-CoV-2 Infections. No other potential conflicts of interest were disclosed.

Use of molecular data in human and ecological health risk assessments of industrial chemicals and agrochemicals has been anticipated by the scientific community for many years; however, these data are rarely used for risk assessment. Here, a logic framework is proposed to explore the feasibility and future development of transcriptomic methods to refine and replace the current apical endpoint-based regulatory toxicity testing paradigm. Four foundational principles are outlined and discussed that would need to be accepted by stakeholders prior to this transformative vision being realized. Well-supported by current knowledge, the first principle is that transcriptomics is a reliable tool for detecting alterations in gene expression that result from endogenous or exogenous influences on the test organism. The second principle states that alterations in gene expression are indicators of adverse or adaptive biological responses to stressors in an organism. Principle 3 is that transcriptomics can be employed to establish a benchmark dose-based point of departure (POD) from short-term, in vivo studies at a dose level below which a concerted molecular change (CMC) is not expected. Finally, Principle 4 states that the use of a transcriptomic POD (set at the CMC dose level) will support a human health-protective risk assessment. If all four principles are substantiated, this vision is expected to transform aspects of the industrial chemical and agrochemical risk assessment process that are focused on establishing safe exposure levels for mammals across numerous toxicological contexts resulting in a significant reduction in animal use while providing equal or greater protection of human health. Importantly, these principles and approaches are also generally applicable for ecological safety assessment., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society of Toxicology.)

Background: Keratinocytes constitute a major part of the melanoma microenvironment, considering their protective role towards melanocytes in physiological conditions. However, their interactions with tumor cells following melanomagenesis are still unclear., Methods: We used two in vitro models (melanoma-conditioned media and indirect co-culture of keratinocytes with melanoma cells on Transwell inserts) to activate immortalized keratinocytes towards cancer-associated ones. Western Blotting and qPCR were used to evaluate keratinocyte markers and mediators of cell invasiveness on protein and mRNA expression level respectively. The levels and activity of proteases and cytokines were analysed using gelatin-FITC staining, gelatin zymography, chemiluminescent enzymatic test, as well as protein arrays. Finally, to further study the functional changes influenced by melanoma we assessed the rate of proliferation of keratinocytes and their invasive abilities by employing wound healing assay and the Transwell filter invasion method., Results: HaCaT keratinocytes activated through incubation with melanoma-conditioned medium or indirect co-culture exhibit properties of less differentiated cells (downregulation of cytokeratin 10), which also prefer to form connections with cancer cells rather than adjacent keratinocytes (decreased level of E-cadherin). While they express only a small number of cytokines, the variety of secreted proteases is quite prominent especially considering that several of them were never reported as a part of secretome of activated keratinocytes' (e.g., matrix metalloproteinase 3 (MMP3), ADAM metallopeptidase with thrombospondin type 1 motif 1). Activated keratinocytes also seem to exhibit a high level of proteolytic activity mediated by MMP9 and MMP14, reduced expression of TIMPs (tissue inhibitor of metalloproteinases), upregulation of ERK activity and increased levels of MMP expression regulators-RUNX2 and galectin 3. Moreover, cancer-associated keratinocytes show slightly elevated migratory and invasive abilities, however only following co-culture with melanoma cells on Transwell inserts., Conclusions: Our study offers a more in-depth view of keratinocytes residing in the melanoma niche, drawing attention to their unique secretome and mediators of invasive abilities, factors which could be used by cancer cells to support their invasion of surrounding tissues. Video abstract., (© 2022. The Author(s).)

Antibody decline occurred from 2 weeks to 6 months post-BNT162b2 mRNA vaccination in nursing home (NH) residents and healthcare workers. Antispike, receptor-binding domain, and neutralization levels dropped >81% irrespective of prior infection. Notably, 69% of infection-naive NH residents had neutralizing antibodies at or below the assay's limit of detection., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America.)

Background: Nursing home (NH) residents have borne a disproportionate share of SARS-CoV-2 morbidity and mortality. Vaccines have limited hospitalisation and death from earlier variants in this vulnerable population. With the rise of Omicron and future variants, it is vital to sustain and broaden vaccine-induced protection. We examined the effect of boosting with BNT162b2 mRNA vaccine on humoral immunity and Omicron-specific neutralising activity among NH residents and healthcare workers (HCWs)., Methods: We longitudinally enrolled 85 NH residents (median age 77) and 48 HCWs (median age 51), and sampled them after the initial vaccination series; and just before and 2 weeks after booster vaccination. Anti-spike, anti-receptor binding domain (RBD) and neutralisation titres to the original Wuhan strain and neutralisation to the Omicron strain were obtained., Findings: Booster vaccination significantly increased vaccine-specific anti-spike, anti-RBD, and neutralisation levels above the pre-booster levels in NH residents and HCWs, both in those with and without prior SARS-CoV-2 infection. Omicron-specific neutralisation activity was low after the initial 2 dose series with only 28% of NH residents' and 28% HCWs' titres above the assay's lower limit of detection. Omicron neutralising activity following the booster lifted 86% of NH residents and 93% of HCWs to the detectable range., Interpretation: With boosting, the vast majority of HCWs and NH residents developed detectable Omicron-specific neutralising activity. These data provide immunologic evidence that strongly supports booster vaccination to broaden neutralising activity and counter waning immunity in the hope it will better protect this vulnerable, high-risk population against the Omicron variant., Funding: NIH AI129709-03S1, U01 CA260539-01, CDC 200-2016-91773, and VA BX005507-01., (Published by Elsevier B.V.)

After BNT162b2 messenger RNA vaccination, antibody levels to spike, receptor-binding domain, and virus neutralization were examined in 149 nursing home residents and 110 healthcare worker controls. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-naive nursing home residents' median post-second vaccine dose antibody neutralization titers are one-quarter that of SARS-CoV-2-naive healthcare workers., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America.)

Background: The BNT162b2 SARS-CoV-2 mRNA vaccination has mitigated the burden of COVID-19 among residents of long-term care facilities considerably, despite being excluded from the vaccine trials. Data on reactogenicity (vaccine side effects) in this population are limited., Aims: To assess reactogenicity among nursing home (NH) residents. To provide a plausible proxy for predicting vaccine response among this population., Methods: We enrolled and sampled NH residents and community-dwelling healthcare workers who received the BNT162b2 mRNA vaccine, to assess local or systemic reactogenicity and antibody levels (immunogenicity)., Results: NH residents reported reactions at a much lower frequency and lesser severity than the community-dwelling healthcare workers. These reactions were mild and transient with all subjects experiencing more local than systemic reactions. Based on our reactogenicity and immunogenicity data, we developed a linear regression model predicting log-transformed anti-spike, anti-receptor-binding domain (RBD), and neutralizing titers, with a dichotomous variable indicating the presence or absence of reported reactions which revealed a statistically significant effect, with estimated shifts in log-transformed titers ranging from 0.32 to 0.37 (all p < 0.01) indicating greater immunogenicity in subjects with one or more reported reactions of varying severity., Discussion: With a significantly lower incidence of post-vaccination reactions among NH residents as reported in this study, the BNT162b2 mRNA vaccine appears to be well-tolerated among this vulnerable population. If validated in larger populations, absence of reactogenicity could help guide clinicians in prioritizing vaccine boosters., Conclusions: Reactogenicity is significantly mild among nursing home residents and overall, subjects who reported post-vaccination reactions developed higher antibody titers., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Provision of nutrition care is vital to the health and well-being of any patient who enters the health care system, whether in the ambulatory, inpatient, or long-term care setting. Interdisciplinary professionals-nurses, physicians, advanced practice providers, pharmacists, and dietitians-identify and treat nutrition problems or clinical conditions in each of these health care settings. The documentation of nutrition care in a structured format from screening and assessment to discharge allows communication of the nutrition treatment plans. The goal of this document is to provide recommendations to clinicians for working with an organization's Information Systems department to create tools for documentation of nutrition care in the electronic health record. These recommendations can also serve as guidance for health care organizations choosing and implementing health care software., (Copyright © 2020 Academy of Nutrition and Dietetics, American Society for Parenteral and Enteral Nutrition, and Association of Clinical Documentation Improvement Specialists. Published by Elsevier Inc. All rights reserved.)

Introduction: The aim of our study was to evaluate testicular growth and risk of atrophy at different lengths of time from the 2SLF-SO in boys with intra-abdominal cryptorchidism., Material and Methods: Twenty-seven boys aged one to 24 months were treated for 35 non-palpable testes. Twenty-one boys with 29 IAT underwent a two-stage laparoscopic Fowler-Stephens operation. The patients underwent ultrasound examination of the TV at six time points: before treatment (TP1), 3 months (TP2) and 9-12 months after laparoscopic F-S operation (TP3), 3 months after inguinal orchidopexy (TP4), 3-6 years of age (TP5) and 7-9 years of age (TP6). The testicular atrophy index (TAI) of the affected testicle was calculated., Results: The overall success rate of treatment was 93.1% (27/29). The median volume of the affected testicles before treatment was less than the median TV of the healthy gonad, but increased systematically at the successive time points, showing significant growth between the second and sixth time points ( p < 0.02, R = 2.75). The median TAI value decreased at the subsequent time points from the level of 26% at TP1 to 5.4% at TP6, with no significant difference., Conclusions: Two-stage laparoscopic Fowler-Stevens operation proved to be an effective procedure for the treatment of intra-abdominal testicles. The undescended testis had a chance to grow and to equalize with the healthy testis after this procedure. The incidence of testicular atrophy was low., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2019 Termedia & Banach.)

Introduction: Colorectal cancer is the second most common cancer worldwide. DNA double strand breaks (DSBs) are the most dangerous lesions which can lead to carcinogenesis. Nonhomologous end joining (NHEJ) is an important pathway, that allows for recovering DNA by direct end joining. The XRCC6 and LIG4 genes encode respectively Ku70 protein and human ATP-dependent DNA ligase, which are the components of the NHEJ repair pathway. The aim of our study was to evaluate the influence of XRCC6 C1310G and LIG4 T9I genes polymorphisms on colorectal cancer risk among Polish population., Materials and Method: Genotyping was performed using TaqMan probes based on analysis of PCR products amplified in Real Time PCR. The research has been carried out on the material obtained from 100 patients with colorectal cancer and 100 cancer-free individuals who were age and sex-matched as a control group. The results were developed using the chi - squer test and odds ratio (OR)., Results: Odd ratio analysis indicates reduced risk of colorectal cancer for LIG4 T9I polymorphism in heterozygotus model C/T (OR= 0.2717 95% CI= 0.1247-0,5918) and homozygous model T/T (OR= 0.3593 95% CI= 0.1394-0.9266). Similar situation we observed for XRCC6 C1310G gene polymorphism, which indicated on heterozygotus variant C/G (OR= 0.1181 95% CI= 0.0145-0.964) and homozygotus variant G/G (OR= 0.0972 95% CI= 0.0097-0.9713) to decrease the risk of colorectal cancer. Conslusions: Our research revealed XRCC6 C1310G and LIG4 T9I polymorphisms are associated with diminished risk of colorectal cancer. However, to confirm obtained results, a further investigations should be carried out.

ADJUST, a novel ankle-foot orthosis (AFO) that we have developed, allows the ankle a normal range of motion (ROM) while providing support for flaccid ankle-muscle paresis. It consists of two leaf-spring hinges that independently control plantarflexion and dorsiflexion stiffness. To evaluate whether ADJUST meets the minimum mechanical requirements, we quantified its ankle ROM and stiffness. To evaluate whether it meets the minimum ankle kinematic and kinetic goals for normal gait, a patient with both plantarflexor and dorsiflexor paralysis used it, and his own AFO, to walk. When fitted with stiff springs, ADJUST met all requirements and goals. During the stance and the swing phases, ankle ROM was within the normal range when ADJUST was fitted with stiff springs. Ankle ROM during stance was outside the normal range both with the patient's own AFO and with ADJUST when it was fitted with flexible springs. Power at the ankle met the minimum goal but was lower with ADJUST than with the patient's own AFO. The optimal stiffness configuration that would result in a higher power at the ankle with a normal ankle ROM was not reached for this patient. Walking with ADJUST seems feasible and could be profitable in patients with flaccid ankle muscle paresis.

Background: Improving ankle-foot orthosis design can best be done by implementing a user-centered approach., Objective: To provide insight into the ideas of ankle-foot orthosis users with flaccid ankle muscle paresis on the importance of activities and suggestions for an improved ankle-foot orthosis design., Study Design: A focus-group discussion with eight ankle-foot orthosis users (57 ± 5 years, 50% female)., Methods: Main inclusion criteria were as follows: ⩾18 years, unable to stand on tip-toe and unable to lift toes. Main exclusion criterion was spasticity of lower extremity muscles. Transcribed data were coded according to the International Classification of Functioning, Disability and Health. Thematic analysis with inductive approach was chosen to order and interpret codes., Results: Ankle-foot orthosis users ranked walking the most important activity followed by sitting down/standing up from a chair. Their opinion was that ankle-foot orthoses facilitate walking and standing. Ankle-foot orthosis users suggested that an improved ankle-foot orthosis design should balance between stability and flexibility., Conclusion: Current ankle-foot orthoses facilitate walking which was the most important activity according to ankle-foot orthosis users. An improved ankle-foot orthosis design should enable walking and should optimize between stability and flexibility dependent on the activity and the paresis severity. Clinical relevance Experienced users of ankle-foot orthosis agreed that matching ankle-foot orthosis functions to daily-life activities is a trade-off between stability and flexibility. An improved ankle-foot orthosis design should at least enable level walking.

We have proposed that safe and effective protection against the development of adult onset cancers may be achieved by vaccination against tissue-specific self-proteins that are "retired" from expression at immunogenic levels in normal tissues as we age, but are overexpressed in emerging tumors. α-Lactalbumin is an example of a "retired" self-protein because its expression in normal tissues is confined exclusively to the breast during late pregnancy and lactation, but is also expressed in the vast majority of human triple negative breast cancers (TNBC)-the most aggressive and lethal form of breast cancer and the predominant form that occurs in women at high genetic risk including those with mutated BRCA1 genes. In anticipation of upcoming clinical trials, here we provide preclinical data indicating that α-lactalbumin has the potential as a vaccine target for inducing safe and effective primary immunoprevention as well as immunotherapy against TNBC.

Background: People with floppy ankle muscles paresis use ankle foot orthoses to improve their walking ability. Ankle foot orthoses also limit ankle range of motion thereby introducing additional problems. Insight in effects of ankle foot orthoses on body functions and activities in people with floppy paretic ankle muscles aids in clinical decision making and may improve adherence., Methods: Studies published before October 27th, 2014, were searched in Pubmed, Embase, Cinahl, and Cochrane Library. Studies evaluating effects of ankle foot orthoses on body functions and/or activities in people with floppy paretic ankle muscles were included. Studies solely focusing on people with spastic paretic ankle muscles were excluded. Study quality was assessed using a custom-made scale. Body functions and activities were defined according to the International Classification of Functioning, Disability and Health., Findings: Twenty-four studies were included, evaluating 394 participants. Participants were grouped according to paresis type (i) dorsiflexor paresis, (ii) plantar flexor paresis, (iii) both dorsiflexor and plantar flexor paresis. Dorsal, circular, and elastic ankle foot orthoses increased dorsiflexion during swing (by 4-6°, group i). Physical comfort with dorsal ankle foot orthoses was lower than that with circular ankle foot orthoses (groups i and iii). Dorsal ankle foot orthoses increased push-off moment (by 0.2-0.5 Nm/kg), increased walking efficiency, and decreased ankle range of motion (by 12-30°, groups ii and iii)., Interpretation: People with dorsiflexor paresis benefit more from circular and elastic ankle foot orthoses while people with plantar flexor paresis (and dorsiflexor paresis) benefit more from dorsal ankle foot orthoses., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Purpose: To evaluate how accurately final infarct volume in acute ischemic stroke can be predicted with perfusion CT (PCT) using a 64-MDCT unit and the toggling table technique., Materials and Methods: Retrospective analysis of 89 patients with acute ischemic stroke who underwent CCT, CT angiography (CTA) and PCT using the “toggling table” technique within the first three hours after symptom onset. In patients with successful thrombolytic therapy (n = 48) and in those without effective thrombolytic therapy (n = 41), the infarct volume and the volume of the penumbra on PCT were compared to the infarct size on follow-up images (CT or MRI) performed within 8 days. The feasibility of complete infarct volume prediction by 8 cm cranio-caudal coverage was evaluated., Results: The correlation between the volume of hypoperfusion on PCT defined by cerebral blood volume reduction and final infarct volume was strongest in patients with successful thrombolytic therapy with underestimation of the definite infarct volume by 8.5 ml on average. The CBV map had the greatest prognostic value. In patients without successful thrombolytic therapy, the final infarct volume was overestimated by 12.1 ml compared to the MTT map on PCT. All infarcts were detected completely. There were no false-positive or false-negative results., Conclusion: Using PCT and the “toggling table” technique in acute stroke patients is helpful for the rapid and accurate quantification of the minimal final infarct and is therefore a prognostic parameter which has to be evaluated in further studies to assess its impact on therapeutic decision.