Validating the OCS-Plus against a clinical standard: A brief report.

This research aimed to determine the sensitivity and clinical validity of the OCS-Plus, a stroke-specific tablet-based cognitive screening tool, in comparison with the MoCA, a routinely used screening tool, after stroke. Eighty-six patients were recruited from Oxfordshire stroke wards over a 22-month period and completed both screens. Overall, we found that the OCS-Plus has good convergent validity and excellent sensitivity when compared with the MoCA. The OCS-Plus is therefore of potential benefit to those seeking a sensitive screening tool.

ß-amyloid accumulation enhances microtubule associated protein tau pathology in an APPNL-G-F/MAPTP301S mouse model of Alzheimer's disease.

Introduction: The study of the pathophysiology study of Alzheimer's disease (AD) has been hampered by lack animal models that recapitulate the major AD pathologies, including extracellular -amyloid (A) deposition, intracellular aggregation of microtubule associated protein tau (MAPT), inflammation and neurodegeneration. Methods: The humanized APPNL-G-F knock-in mouse line was crossed to the PS19 MAPTP301S, over-expression mouse line to create the dual APPNL-G-F/PS19 MAPTP301S line. The resulting pathologies were characterized by immunochemical methods and PCR. Results: We now report on a double transgenic APPNL-G-F/PS19 MAPTP301S mouse that at 6 months of age exhibits robust A plaque accumulation, intense MAPT pathology, strong inflammation and extensive neurodegeneration. The presence of A pathology potentiated the other major pathologies, including MAPT pathology, inflammation and neurodegeneration. MAPT pathology neither changed levels of amyloid precursor protein nor potentiated A accumulation. Interestingly, study of immunofluorescence in cleared brains indicates that microglial inflammation was generally stronger in the hippocampus, dentate gyrus and entorhinal cortex, which are regions with predominant MAPT pathology. The APPNL-G-F/MAPTP301S mouse model also showed strong accumulation of N6-methyladenosine (m6A), which was recently shown to be elevated in the AD brain. m6A primarily accumulated in neuronal soma, but also co-localized with a subset of astrocytes and microglia. The accumulation of m6A corresponded with increases in METTL3 and decreases in ALKBH5, which are enzymes that add or remove m6A from mRNA, respectively. Discussion: Our understanding of the pathophysiology of Alzheimer's disease (AD) has been hampered by lack animal models that recapitulate the major AD pathologies, including extracellular -amyloid (A) deposition, intracellular aggregation of microtubule associated protein tau (MAPT), inflammation and neurodegeneration. The APPNL-G-F/MAPTP301S mouse recapitulates many features of AD pathology beginning at 6 months of aging, and thus represents a useful new mouse model for the field.

Frequency of Leaks from Conical Centrifuge Tubes.

Introduction: Threaded conical centrifuge tubes are ubiquitous in biological laboratories and are frequently used for the storage/transport of potentially biohazardous samples. However, limited data are available on how frequently and from where these tubes leak. These data are valuable for laboratory biorisk management and to inform future studies on risks arising from the routine use of laboratory consumables. Methods: The frequency of leaks from threaded conical centrifuge tubes was tested using a Glo Germ solution as a tracer. Conical tubes (15 and 50 mL) from several brands were filled, inverted, and placed on their side on the benchtop. After 1 h, the presence or absence of leaks on the benchtop surface, tube threads, and exterior was recorded. Results: We observed that liquid leaked out of tubes that were apparently properly threaded in 2% of 15 mL tubes (confidence interval [95% CI] 1.4-2.6) and 1.4% of 50 mL tubes (95% CI 0.2-1.5). After opening, liquid was found on the threads on the outside of the tube in 20% of 15 mL tubes (95% CI 10-31) and 14% of 50 mL tubes (95% CI 1-28). We did not find sufficient evidence that differences in leak rates among brands were practically significant. Conclusions: The fact that leaks were not uncommonly observed from conical centrifuge tubes suggests that mitigations for any hazard posed by a leak should be a component of every biorisk management strategy for protocols involving the manipulation of hazardous substances in these tubes. Further research should be conducted on other activities that could cause tubes to leak (such as centrifugation or vortexing) and should be completed to understand the risks associated with this consumable. Research into the costs and benefits of mitigating the risk of leaks from conical tubes is recommended.

Online monitoring of higher alcohols and esters throughout beer fermentation by commercial Saccharomyces cerevisiae and Saccharomyces pastorianus yeast.

Higher alcohols and esters are among the predominant classes of volatile organic compounds (VOCs) that influence the quality of beer. The concentrations of these compounds are determined through a specific yeast strain selection and fermentation conditions. The effect of yeast strains on the formation of higher alcohols and esters throughout fermentations (at 20°C) was investigated. Flavour-relevant esters (ethyl acetate, isoamyl acetate, ethyl hexanoate and ethyl octanoate) and higher alcohols (isoamyl alcohol, isobutyl alcohol and phenylethyl alcohol) were monitored throughout the fermentation using proton transfer reaction-time of flight-mass spectrometry (PTR-ToF-MS) coupled with an automated sampling system for continuous measurements. Compound identification was confirmed by analysis of samples using gas chromatography-mass spectrometry (GC-MS). Results demonstrated the specific time points where variation in higher alcohol and ester generation between yeast strains occurred. In particular, the concentrations of isoamyl acetate, ethyl octanoate and isoamyl alcohol between yeast strains were significantly different over the first 2 days of fermentation; whereas, after Day 3, no significant differences were observed. The two Saccharomyces pastorianus strains produced comparable concentrations of the key higher alcohols and esters. However, the key higher alcohol and ester concentrations varied greatly between the two S. cerevisiae strains. The use of PTR-ToF-MS to rapidly measure multiple yeast strains provides new insights on fermentation for brewers to modify the sensory profile and optimise quality.

Curriculum learning-based strategy for low-density archaeological mound detection from historical maps in India and Pakistan.

This paper presents two algorithms for the large-scale automatic detection and instance segmentation of potential archaeological mounds on historical maps. Historical maps present a unique source of information for the reconstruction of ancient landscapes. The last 100 years have seen unprecedented landscape modifications with the introduction and large-scale implementation of mechanised agriculture, channel-based irrigation schemes, and urban expansion to name but a few. Historical maps offer a window onto disappearing landscapes where many historical and archaeological elements that no longer exist today are depicted. The algorithms focus on the detection and shape extraction of mound features with high probability of being archaeological settlements, mounds being one of the most commonly documented archaeological features to be found in the Survey of India historical map series, although not necessarily recognised as such at the time of surveying. Mound features with high archaeological potential are most commonly depicted through hachures or contour-equivalent form-lines, therefore, an algorithm has been designed to detect each of those features. Our proposed approach addresses two of the most common issues in archaeological automated survey, the low-density of archaeological features to be detected, and the small amount of training data available. It has been applied to all types of maps available of the historic 1″ to 1-mile series, thus increasing the complexity of the detection. Moreover, the inclusion of synthetic data, along with a Curriculum Learning strategy, has allowed the algorithm to better understand what the mound features look like. Likewise, a series of filters based on topographic setting, form, and size have been applied to improve the accuracy of the models. The resulting algorithms have a recall value of 52.61% and a precision of 82.31% for the hachure mounds, and a recall value of 70.80% and a precision of 70.29% for the form-line mounds, which allowed the detection of nearly 6000 mound features over an area of 470,500 km2, the largest such approach to have ever been applied. If we restrict our focus to the maps most similar to those used in the algorithm training, we reach recall values greater than 60% and precision values greater than 90%. This approach has shown the potential to implement an adaptive algorithm that allows, after a small amount of retraining with data detected from a new map, a better general mound feature detection in the same map.

Accumulation of m6A exhibits stronger correlation with MAPT than ß-amyloid pathology in an APPNL-G-F /MAPTP301S mouse model of Alzheimer's disease.

The study for the pathophysiology study of Alzheimer's disease (AD) has been hampered by lack animal models that recapitulate the major AD pathologies, including extracellular ß-amyloid (Aß) deposition, intracellular aggregation of microtubule associated protein tau (MAPT), inflammation and neurodegeneration. We now report on a double transgenic APPNL-G-F MAPTP301S mouse that at 6 months of age exhibits robust Aß plaque accumulation, intense MAPT pathology, strong inflammation and extensive neurodegeneration. The presence of Aß pathology potentiated the other major pathologies, including MAPT pathology, inflammation and neurodegeneration. However, MAPT pathology neither changed levels of amyloid precursor protein nor potentiated Aß accumulation. The APPNL-G-F/MAPTP301S mouse model also showed strong accumulation of N6-methyladenosine (m6A), which was recently shown to be elevated in the AD brain. M6A primarily accumulated in neuronal soma, but also co-localized with a subset of astrocytes and microglia. The accumulation of m6A corresponded with increases in METTL3 and decreases in ALKBH5, which are enzymes that add or remove m6A from mRNA, respectively. Thus, the APPNL-G-F/MAPTP301S mouse recapitulates many features of AD pathology beginning at 6 months of aging.

Rate of Splashes When Opening Microfuge Tubes with Various Methods.

Introduction: Snap-cap microcentrifuge tubes are ubiquitous in biological laboratories. However, limited data are available on how frequently splashes occur when opening them. These data would be valuable for biorisk management in the laboratory. Methods: The frequency of splashes from opening snap-cap tubes using four different methods was tested. The splash frequency for each method was measured on the benchtop surface and on the experimenter's gloves and smock, using a Glo Germ solution as a tracer. Results: Splashes occurred very frequently when opening microcentrifuge snap-cap tubes, no matter which method was used to open the tube. The highest rate of splashes on all surfaces was observed with the one-handed (OH) opening method compared with two-handed methods. Across all methods, the highest rate of splashes was observed on the opener's gloves (70-97%) compared with the benchtop (2-40%) or the body of the researcher (0-7%). Conclusions: All tube opening methods we studied frequently caused splashes, with the OH method being the most error-prone but no two-handed method being clearly superior to any other. In addition to posing an exposure risk to laboratory personnel, experimental repeatability may be affected due to loss of volume when using snap-cap tubes. The rate of splashes underscores the importance of secondary containment, personal protective equipment, and good protocols for decontamination. When working with especially hazardous materials, alternatives to snap-cap tubes (such as screw cap tubes) should be strongly considered. Future studies can examine other methods of opening snap-cap tubes to determine whether a truly safe method exists.

Accumulation of m6A exhibits stronger correlation with MAPT than ß-amyloid pathology in an APPNL-G-F /MAPTP301S mouse model of Alzheimer's disease.

The study for the pathophysiology study of Alzheimer's disease (AD) has been hampered by lack animal models that recapitulate the major AD pathologies, including extracellular ß-amyloid (Aß) deposition, intracellular aggregation of microtubule associated protein tau (MAPT), inflammation and neurodegeneration. We now report on a double transgenic APPNL-G-F MAPTP301S mouse that at 6 months of age exhibits robust Aß plaque accumulation, intense MAPT pathology, strong inflammation and extensive neurodegeneration. The presence of Aß pathology potentiated the other major pathologies, including MAPT pathology, inflammation and neurodegeneration. However, MAPT pathology neither changed levels of amyloid precursor protein nor potentiated Aß accumulation. The APPNL-G-F/MAPTP301S mouse model also showed strong accumulation of N6-methyladenosine (m6A), which was recently shown to be elevated in the AD brain. M6A primarily accumulated in neuronal soma, but also co-localized with a subset of astrocytes and microglia. The accumulation of m6A corresponded with increases in METTL3 and decreases in ALKBH5, which are enzymes that add or remove m6A from mRNA, respectively. Thus, the APPNL-G-F/MAPTP301S mouse recapitulates many features of AD pathology beginning at 6 months of aging.

One size does not fit all: Impact of hand size on ease of use of instruments for minimally invasive surgery.

BACKGROUND: Consideration of ergonomic factors is important for the practice of safe and efficient minimally invasive surgery (MIS). Surgeons with smaller glove sizes have previously been reported to have increased difficulties with some minimally invasive instruments. We aim to investigate hand anthropometrics and their relationship to surgeon comfort when using MIS instruments. METHODS: Male and female surgeons from two centres were surveyed on their experience of handling MIS instruments and images obtained of the dorsal and palmar aspects of their dominant hand. Photographs of hands were transformed to calibrated coordinates to enable anthropometric measurements of finger length and width as well as palm width and hand span photogrammetrically. Surgeon-perceived discomfort, fatigue, pressure points and techniques to mitigate difficulty handling instruments were compared to hand measurements. RESULTS: Questionnaires were completed by 58 surgeons; 20 (34%) were consultants, 17 (29%) were women. Glove size ranged from 6 to 8 (median 7.5). Male participants had significantly larger hands than females in all measured dimensions. Female surgeons and those with smaller finger and hand dimensions were significantly more likely to experience difficulty or discomfort across a range of variables when using MIS instruments. CONCLUSIONS: Surgeons with smaller hands reported increased problems handling MIS instruments. This represents an issue of equity in surgery, with women being more significantly affected than men. Hand size varies greatly between surgeons and anthropometric variability should be considered in design of MIS instruments.

Developmental changes in colour constancy in a naturalistic object selection task.

When the illumination falling on a surface change, so does the reflected light. Despite this, adult observers are good at perceiving surfaces as relatively unchanging-an ability termed colour constancy. Very few studies have investigated colour constancy in infants, and even fewer in children. Here we asked whether there is a difference in colour constancy between children and adults; what the developmental trajectory is between six and 11 years; and whether the pattern of constancy across illuminations and reflectances differs between adults and children. To this end, we developed a novel, child-friendly computer-based object selection task. In this, observers saw a dragon's favourite sweet under a neutral illumination and picked the matching sweet from an array of eight seen under a different illumination (blue, yellow, red, or green). This set contained a reflectance match (colour constant; perfect performance) and a tristimulus match (colour inconstant). We ran two experiments, with two-dimensional scenes in one and three-dimensional renderings in the other. Twenty-six adults and 33 children took part in the first experiment; 26 adults and 40 children took part in the second. Children performed better than adults on this task, and their performance decreased with age in both experiments. We found differences across illuminations and sweets, but a similar pattern across both age groups. This unexpected finding might reflect a real decrease in colour constancy from childhood to adulthood, explained by developmental changes in the perceptual and cognitive mechanisms underpinning colour constancy, or differences in task strategies between children and adults. HIGHLIGHTS: Six- to 11-year-old children demonstrated better performance than adults on a colour constancy object selection task. Performance decreased with age over childhood. These findings may indicate development of cognitive strategies used to overcome automatic colour constancy mechanisms.

Using machine learning to examine drivers of inappropriate outpatient antibiotic prescribing in acute respiratory illnesses.

Using a machine-learning model, we examined drivers of antibiotic prescribing for antibiotic-inappropriate acute respiratory illnesses in a large US claims data set. Antibiotics were prescribed in 11% of the 42 million visits in our sample. The model identified outpatient setting type, patient age mix, and state as top drivers of prescribing.

Pandemic Stories: The Voices of Older Adults.

To document the reactions and experiences of older persons during the COVID-19 pandemic, we have conducted a qualitative study of 25 older adults (50 years of age and older) throughout the pandemic, using serial interviewing methods. This analysis reports on the data collected from the first two rounds of interviews - one conducted in the summer of 2020 and one conducted in the fall of 2020. Our thematic analysis found eight major themes: thoughts on the dangers of the pandemic, how the virus has changed daily life (including social life), health care during COVID (being a caregiver, losing a loved one, seeking health care), missing spontaneity and dealing with existential dread, the growing frustration, seeking connection through civic participation, adaptation and resilience, and the social ills that the pandemic has revealed. These stories describe both loneliness and connection, hope coupled with disappointment, but overwhelmingly, an insight into what the pandemic has shown us about the social ills that it has revealed.

Validation of the UK English Oxford cognitive screen-plus in sub-acute and chronic stroke survivors.

Introduction: Stroke survivors are routinely screened for cognitive impairment with tools that often fail to detect subtle impairments. The Oxford Cognitive Screen-Plus (OCS-Plus) is a brief tablet-based screen designed to detect subtle post-stroke cognitive impairments. We examined its psychometric properties in two UK English-speaking stroke cohorts (subacute: <3 months post-stroke, chronic: >6 months post-stroke) cross-sectionally. Patients and methods: This study included 347 stroke survivors (mean age = 73 years; mean education = 13 years; 43.06% female; 74.42% ischaemic stroke). The OCS-Plus was completed by 181 sub-acute stroke survivors and 166 chronic stroke survivors. All participants also completed the Oxford Cognitive Screen (OCS) and a subset completed the Montreal Cognitive Assessment (MoCA) and further neuropsychological tests. Results: First, convergent construct validity of OCS-Plus tasks to task-matched standardized neuropsychological tests was confirmed (r > 0.30). Second, we evaluated divergent construct validity of all OCS-Plus subtasks (r < 0.19). Third, we report the sensitivity and specificity of each OCS-Plus subtask compared to neuropsychological test performance. Fourth, we found that OCS-Plus detected cognitive impairments in a large proportion of those classed as unimpaired on MoCA (100%) and OCS (98.50%). Discussion and conclusion: The OCS-Plus provides a valid screening tool for sensitive detection of subtle cognitive impairment in stroke patients. Indeed, the OCS-Plus detected subtle cognitive impairment at a similar level to validated neuropsychological assessments and exceeded detection of cognitive impairment compared to standard clinical screening tools.

Spatial and socioeconomic inequities in liveability in Australia's 21 largest cities: Does city size matter?

Spatial and area-level socioeconomic variation in urban liveability (access to social infrastructure, public transport, open space, healthy food choices, local employment, street connectivity, dwelling density, and housing affordability) was examined and mapped across 39,967 residential statistical areas in Australia's metropolitan (n = 7) and largest regional cities (n = 14). Urban liveability varied spatially, with inner-city areas more liveable than outer suburbs. Disadvantaged areas in larger metropolitan cities were less liveable than advantaged areas, but this pattern was reversed in smaller cities. Local data could inform policies to redress inequities, including those designed to avoid disadvantage being suburbanised as cities grow and gentrify.

Odorant receptor orthologues in conifer-feeding beetles display conserved responses to ecologically relevant odours.

Insects are able to detect a plethora of olfactory cues using a divergent family of odorant receptors (ORs). Despite the divergent nature of this family, related species frequently express several evolutionarily conserved OR orthologues. In the largest order of insects, Coleoptera, it remains unknown whether OR orthologues have conserved or divergent functions in different species. Using HEK293 cells, we addressed this question through functional characterization of two groups of OR orthologues in three species of the Curculionidae (weevil) family, the conifer-feeding bark beetles Ips typographus L. ("Ityp") and Dendroctonus ponderosae Hopkins ("Dpon") (Scolytinae), and the pine weevil Hylobius abietis L. ("Habi"; Molytinae). The ORs of H. abietis were annotated from antennal transcriptomes. The results show highly conserved response specificities, with one group of orthologues (HabiOR3/DponOR8/ItypOR6) responding exclusively to 2-phenylethanol (2-PE), and the other group (HabiOR4/DponOR9/ItypOR5) responding to angiosperm green leaf volatiles (GLVs). Both groups of orthologues belong to the coleopteran OR subfamily 2B, and share a common ancestor with OR5 in the cerambycid Megacyllene caryae, also tuned to 2-PE, suggesting a shared evolutionary history of 2-PE receptors across two beetle superfamilies. The detected compounds are ecologically relevant for conifer-feeding curculionids, and are probably linked to fitness, with GLVs being used to avoid angiosperm nonhost plants, and 2-PE being important for intraspecific communication and/or playing a putative role in beetle-microbe symbioses. To our knowledge, this study is the first to reveal evolutionary conservation of OR functions across several beetle species and hence sheds new light on the functional evolution of insect ORs.

Physical Activity and Food Environments in and around Schools: A Case Study in Regional North-West Tasmania.

A better understanding of the physical activity (PA) infrastructure in schools, the walkability of neighborhoods close to schools, and the food environments around schools, particularly in rural, socioeconomically challenged areas such as the North-West (NW) of Tasmania, could be important in the wider effort to improve the health of school-age children. Accordingly, this research aimed to assess PA resources, walkability, and food environments in and around schools in three socioeconomically disadvantaged, regional/rural Local Government Areas (LGAs) of Tasmania, Australia. A census of schools (including assessment of the PA infrastructure quality within school grounds), a walkability assessment, and a census of food outlets surrounding schools (through geospatial mapping) were executed. Most of the schools in the study region had access to an oval, basketball/volleyball/netball court, and free-standing exercise equipment. In all instances (i.e., regardless of school type), the quality of the available infrastructure was substantially higher than the number of incivilities observed. Most schools also had good (i.e., within the first four deciles) walkability. Numerous food outlets were within the walking zones of all schools in the study region, with an abundance of food outlets that predominantly sold processed unhealthy food.

The myometrial-cervical ratio (MCR): Assessing the diagnostic accuracy of a novel ultrasound measurement in the diagnosis of adenomyosis.

BACKGROUND: Adenomyosis is histologically defined by the presence of endometrial glands and stroma in the myometrium. To date, there are no standardised ultrasound findings that reliably predict histological adenomyosis. AIMS: This study aimed to determine the diagnostic accuracy of a novel sonographic measurement for adenomyosis, the myometrial-cervical ratio (MCR), when compared with histopathological diagnosis. MATERIALS AND METHODS: A single-centre retrospective study was performed. The MCR was calculated from the pre-operative ultrasound, and histopathology reviewed for each case. Accuracy data were analysed in the form of 2 × 2 tables. The discriminative value of the MCR was summarised with a receiver operator characteristic (ROC) curve. Sub-analysis examined the impact of fibroids, hormonal suppression, menopausal status, parity and indication for surgery. RESULTS: Between 1 January 2016 and 31 December 2018, 982 patients underwent hysterectomy for benign non-obstetric indication and adequate pre-operative ultrasound was available for 260. The MCR demonstrated limited diagnostic ability for adenomyosis (area under the receiver operating characteristic curve (AUROC) 0.58, 95% CI 0.51-0.65). However, when applied to cases with no uterine fibroid included in the MCR calculation (n = 133) there was a strong association between MCR and diagnosis of adenomyosis (odds ratio: 5.79, 95% CI: 2.15, 15.62, P = 0.001) with AUROC for this model 0.68 (95% CI: 0.59, 0.77). At an MCR cut-point of 1.74, sensitivity is 67.16% and specificity is 66.15%, with 66.67% of samples correctly classified. CONCLUSIONS: While diagnostic accuracy was suboptimal, the MCR outperforms traditional ultrasound diagnostic features of adenomyosis. The MCR may offer a simple imaging measurement for adenomyosis.

Undergraduate structural biology education: A shift from users to developers of computation and simulation tools.

The use of theory and simulation in undergraduate education in biochemistry, molecular biology, and structural biology is now common, but the skills students need and the curriculum instructors have to train their students are evolving. The global pandemic and the immediate switch to remote instruction forced instructors to reconsider how they can use computation to teach concepts previously approached with other instructional methods. In this review, we survey some of the curricula, materials, and resources for instructors who want to include theory, simulation, and computation in the undergraduate curriculum. There has been a notable progression from teaching students to use discipline-specific computational tools to developing interactive computational tools that promote active learning to having students write code themselves, such that they view computation as another tool for solving problems. We are moving toward a future where computational skills, including programming, data analysis, visualization, and simulation, will no longer be considered an optional bonus for students but a required skill for the 21st century STEM (Science, Technology, Engineering, and Mathematics) workforce; therefore, all physical and life science students should learn to program in the undergraduate curriculum.

Outbreak of SARS-CoV-2 in a Children's Nursery in the United Kingdom.

BACKGROUND: Early Years' Settings (EYSs) provide childcare and education for children 0-5 years old. They remained fully open in England during the third National lockdown when other educational settings were only open for selected children. EYSs are generally considered to be low-risk settings for transmission of severe acute respiratory syndrome coronavirus 2 (SARS CoV-2). METHODS: An observational study describing a large outbreak of SARS CoV-2 within an EYS in Cambridgeshire, United Kingdom. RESULTS: Overall 45 cases of SARS-CoV-2 were identified; 24 adults (71% of staff members) and 21 children (25% of nursery attendees). One case was identified as the alpha variant (B.1.1.7 [VOC-20-DEC-01]). One staff member became critically unwell. CONCLUSIONS: Transmission of SARS-CoV-2 occurred quickly, with a high attack rate; likely a consequence of a variant with enhanced transmissibility and an inability of the setting to adhere to infection control measures.

Codon Optimization of Insect Odorant Receptor Genes May Increase Their Stable Expression for Functional Characterization in HEK293 Cells.

Insect odorant receptor (OR) genes are routinely expressed in Human Embryonic Kidney (HEK) 293 cells for functional characterization ("de-orphanization") using transient or stable expression. However, progress in this research field has been hampered because some insect ORs are not functional in this system, which may be due to insufficient protein levels. We investigated whether codon optimization of insect OR sequences for expression in human cells could facilitate their functional characterization in HEK293 cells with stable and inducible expression. We tested the olfactory receptor co-receptor (Orco) proteins from the bark beetles Ips typographus ("Ityp") and Dendroctonus ponderosae ("Dpon"), and six ItypORs previously characterized in Xenopus laevis oocytes and/or HEK cells. Western blot analysis indicated that codon optimization yielded increased cellular protein levels for seven of the eight receptors. Our experimental assays demonstrated that codon optimization enabled functional characterization of two ORs (ItypOR25 and ItypOR29) which are unresponsive when expressed from wildtype (non-codon optimized) genes. Similar to previous Xenopus oocyte recordings, ItypOR25 responded primarily to the host/conifer monoterpene (+)-3-carene. ItypOR29 responded primarily to (+)-isopinochamphone and similar ketones produced by fungal symbionts and trees. Codon optimization also resulted in significantly increased responses in ItypOR49 to its pheromone ligand (R)-(-)-ipsdienol, and improved responses to the Orco agonist VUAA1 in ItypOrco. However, codon optimization did not result in functional expression of DponOrco, ItypOR23, ItypOR27, and ItypOR28 despite higher protein levels as indicated by Western blots. We conclude that codon optimization may enable or improve the functional characterization of insect ORs in HEK cells, although this method is not sufficient for all ORs that are not functionally expressed from wildtype genes.