Dilutional Effect of Ethylenediaminetetraacetic Acid on Packed Cell Volume in Healthy Dogs.

Packed cell volume (PCV) is commonly used to assess and monitor red blood cell count in animals, but the results can be altered if inappropriate ratios of anticoagulant/blood are used. The purpose of this study was to determine the effect of ideally filled, overfilled, and underfilled K3 ethylenediaminetetraacetic acid (EDTA) tubes with various volumes of healthy dog blood on centrifuged PCV. Six milliliters of blood was obtained from 94 blood donors each. Initial distribution was injected into two nonheparinized microhematocrit tubes. The remainder was instilled into 1.3 mL K3 EDTA spray-dried tubes as 1.5 mL, 1.3 mL, 0.75 mL, 0.5 mL, and 0.25 mL aliquots. Normality was determined using the D'agostino-Pearson method and by visual examination of histograms. Data were analyzed using a repeated-measures analysis of variance with post hoc testing using Tukey's test. There is a statistically significant decrease in the PCV between all groups with progressive underfilling of tubes (P < .0001). The closest difference is between 1.5 and 1.3 mL (P = .0138). Our study suggested that underfilling K3 EDTA tubes significantly and negatively influences the PCV in healthy dogs. Using underfilled K3 EDTA tubes result in a lower PCV compared with directly filled microhematocrit tubes without anticoagulant.

Newly detected DNA viruses in juvenile nasopharyngeal angiofibroma (JNA) and oral and oropharyngeal squamous cell carcinoma (OSCC/OPSCC).

PURPOSE: Approximately 20% of cancers are estimated to have a viral etiology. We aimed to investigate whether DNA of 8 human parvoviruses [bocavirus 1-4 (HBoV1-4), parvovirus B19 (B19V), protoparvoviruses (bufa-, tusa-, and cutavirus)] and 13 human polyomaviruses (HPyV) can be detected in oropharyngeal and oral cavity squamous cell carcinoma (OPSCC/OSCC), and in juvenile nasopharyngeal angiofibroma (JNA) tissue samples. METHODS: Fresh samples of seven JNA tissues and ten paired tissues of OSCC/OPSCC tumor and adjacent healthy tissues were collected. DNA extraction and real-time PCRs were performed to detect HBoV1-4, B19V, bufa- tusa- and cutavirus, and HPyV genomes. RESULTS: JNA specimens were negative for all parvoviruses tested, whereas one JNA sample was Merkel cell polyomavirus (MCPyV) DNA positive. The OSCC/OPSCC samples were negative for the human protoparvoviruses, HBoV1-4, and all human polyomaviruses, except for one patient that was MCPyV DNA positive in both healthy and tumor tissues. Seven OSCC/OPSCC patients were positive for B19V DNA, three of them in both healthy and cancerous tissues and three in only healthy tissues. Three of the B19V DNA-positive patients harbored viral genotype 1, three genotype 2, and one genotype 3B. CONCLUSIONS: These are the first reports of MCPyV and B19V DNA being detected in JNA and OPSCC. The significance of viral DNA positivity is unclear. B19V DNA is known to remain in the tissues lifelong, however, it is of interest that there are some patients with B19 DNA in healthy tissue, but not in the corresponding cancer tissue.

Air movement affects insulatory values of nests constructed by Old World Warblers.

Avian nests provide a location for incubation, and in many cases rearing of nestlings. These energetically demanding activities have meant that nest insulation has been the subject of many studies but few to date have dealt with how variation among species in nest construction materials could affect nest insulation. This study investigated the insulation of nests constructed by three species of Old Word warblers (Sylviidae), which vary in size and composition. Insulatory values, i.e. difference in cooling rate of temperature loggers placed inside and outside a nest, and internal cooling rates within the nest cup were determined using temperature loggers under still-air and moving-air conditions. Insulatory values determined in still-air conditions of a laboratory were significantly different among nests of the different species but not when the values were determined within the smaller volume of a wind tunnel. Moving-air increased insulatory values by an order of magnitude but also increased internal cooling rates in all species. Insulatory values were positively correlated with the nest base thickness. Moving-air increased the cooling rate of the external temperature logger much more than the internal logger, which inflated the insulatory value of a nest wall in moving-air. Reasons for these results may reflect the thermal properties of the materials individually or in combination. Future testing of nest insulation should be under standard conditions that limit air movement but the role of the nest location in situ should be investigated in future research.

Patients with early-stage oropharyngeal cancer can be identified with label-free serum proteomics.

BACKGROUND: The increasing incidence of oropharyngeal squamous cell carcinoma (OPSCC) is mainly related to human papillomavirus (HPV) infection. As OPSCCs are often diagnosed at an advanced stage, mortality and morbidity remain high. There are no diagnostic biomarkers for early detection of OPSCC. METHODS: Serum from 25 patients with stage I-II OPSCC, and 12 healthy controls, was studied with quantitative label-free proteomics using ultra-definition MSE. Statistical analyses were performed to identify the proteins most reliably distinguishing early-stage OPSCCs from controls. P16 was used as a surrogate marker for HPV. P16-positive and P16-negative tumours were analysed separately. RESULTS: With two or more unique proteins per identification, 176 proteins were quantified. A clear separation between patients with early-stage tumours and controls was seen in principal component analysis. Latent structures discriminant analysis identified 96 proteins, most reliably differentiating OPSCC patients from controls, with 13 upregulated and 83 downregulated proteins in study cases. The set of proteins was studied further with network, pathway and protein-protein interaction analyses, and found to participate in lipid metabolism, for example. CONCLUSIONS: We found a set of serum proteins distinguishing early-stage OPSCC from healthy individuals, and suggest a protein set for further evaluation as a diagnostic biomarker panel for OPSCC.

Influence of chloride on the chronic toxicity of sodium nitrate to Ceriodaphnia dubia and Hyalella azteca.

While it has been well established that increasing chloride concentration in water reduces the toxicity of nitrite to freshwater species, little work has been done to investigate the effect of chloride on nitrate toxicity. We conducted acute and chronic nitrate (as sodium nitrate) toxicity tests with the cladoceran Ceriodaphnia dubia and the amphipod Hyalella azteca (chronic tests only) over a range of chloride concentrations spanning natural chloride levels found in surface waters representative of watersheds of the Great Lakes Region. Chronic nitrate toxicity test results with both crustaceans were variable, with H. azteca appearing to be one of the more sensitive invertebrate species tested and C. dubia being less sensitive. While the variability in results for H. azteca were to an extent related to chloride concentration in test water that was distinctly not the case for C. dubia. We concluded that the chloride dependent toxicity of nitrate is not universal among freshwater crustaceans. An additional sodium chloride chronic toxicity test with the US Lab strain of H. azteca in the present study suggested that when present as predominantly sodium chloride and with relatively low concentrations of other ions, there is a narrow range of chloride concentrations over which this strain is most fit, and within which toxicity test data are reliable.

Altered microheterogeneity at several N-glycosylation sites in OPSCC in constant protein expression conditions.

Protein glycosylation responds sensitively to disease states. It is implicated in every hallmark of cancer and has recently started to be considered as a hallmark itself. Changes in N-glycosylation microheterogeneity are more dramatic than those of protein expression due to the non-template nature of protein glycosylation. This enables their potential use in serum-based diagnostics. Here, we perform glycopeptidomics on serum from patients with oropharyngeal squamous cell carcinoma (OPSCC), compared to controls and comparing between cancers based on etiology (human papilloma virus- positive or negative). Using MS2, we then targeted glycoforms, significantly different between the groups, to identify their glycopeptide compositions. Simultaneously we investigate the same serum proteins, comparing whether N-glycosylation changes reflect protein-level changes. Significant glycoforms were identified from proteins such as alpha-1-antitrypsin (SERPINA1), haptoglobin, and different immunoglobulins. SERPINA1 had glycovariance at 2 N-glycosylation sites, that were up to 35 times more abundant in even early-stage OPSCCs, despite minimal differences between SERPINA1 protein levels between groups. Some identified glycoforms' fold changes (FCs) were in line with serum protein level FCs, others were less abundant in early-stage cancers but with great variance in higher-stage cancers, such as on immunoglobulin heavy constant gamma 2, despite no change in protein levels. Such findings indicate that glycovariant analysis might be more beneficial than proteomic analysis, which is yet to be fruitful in the search for biomarkers. Highly sensitive glycopeptide changes could potentially be used in the future for cancer screening. Additionally, characterizing the glycopeptide changes in OPSCC is valuable in the search for potential therapeutic targets.

Effect of test duration and feeding on relative sensitivity of genetically distinct clades of Hyalella azteca.

The amphipod Hyalella azteca is widely used in ecotoxicology laboratories for the assessment of chemical risks to aquatic environments, and it is a cryptic species complex with a number of genetically distinct strains found in wild populations. While it would be valuable to note differences in contaminant sensitivity among different strains collected from various field sites, those findings would be influenced by acclimation of the populations to local conditions. In addition, potential differences in metabolism or lipid storage among different strains may confound assessment of sensitivity in unfed acute toxicity tests. In the present study, our aim was to assess whether there are genetic differences in contaminant sensitivity among three cryptic provisional species of H. azteca. Therefore, we used organisms cultured under the same conditions, assessed their ability to survive for extended periods without food, and conducted fed and unfed acute toxicity tests with two anions (nitrate and chloride) whose toxicities are not expected to be altered by the addition of food. We found that the three genetically distinct clades of H. azteca had substantially different responses to starvation, and the presence/absence of food during acute toxicity tests had a strong role in determining the relative sensitivity of the three clades. In fed tests, where starvation was no longer a potential stressor, significant differences in sensitivity were still observed among the three clades. In light of these differences in sensitivity, we suggest that ecotoxicology laboratories consider using a provisional species in toxicity tests that is a regionally appropriate surrogate.

Spatial and interspecific patterns in persistent contaminant loads in bighead and silver carp from the Illinois River.

We measured concentrations of selected organohalogens, fluorinated compounds and mercury in whole, ground silver (Hypophthalmichthys molitrix) and bighead (H. nobilis) carp from the Illinois River, Illinois, in 2010 and 2011 to characterize spatial and interspecific patterns of contaminant burdens. Silver carp, which had greater lipid content, tended to have greater concentrations of lipophilic compounds. Concentrations of organohalogens were generally greater in carp from the upper reaches of the river. The halogenated compounds were associated with length and lipid content in silver carp. Bighead carp had greater mercury concentrations than did silver carp; total mercury concentrations were negatively associated with lipid content of bighead carp. Perfluorinated compounds, comprised predominantly of perfluorooctane sulfonic acid, did not vary by species or river reach. Chlordanes and polychlorinated biphenyls were of potential concern with regard to the use of these carp as animal feed additives. Our results indicated that, even though they occupy a lower trophic level than many similarly-sized fish, these carp may accumulate measureable concentrations of organic contaminants.

Influence of Test Method Variables on Sensitivity of Neocloeon Triangulifer to a Reference Toxicant in Short-Term, Effluent Style Evaluations.

Recent literature has demonstrated the sensitivity of mayflies to environmental contaminants. However, to date, there are no methods approved by the US Environmental Protection Agency for using sensitive insects like mayflies in whole-effluent toxicity or receiving water toxicity tests. The parthenogenetic mayfly Neocloeon triangulifer has been shown to be amenable to continuous culture in the laboratory, and methods have been described for its use in both acute and chronic toxicity studies. The goal of the present study was to investigate aspects of N. triangulifer testing and culturing methods that might require adjustment so that they are applicable for testing effluents and receiving waters in a short-term exposure. To this end, the influence of organism age, test duration, and test temperature on sensitivity to NaCl as a reference toxicant were tested (concentrations ranging from 182 to 2489 mg/L). Further studies were conducted to assess the utility of commercially available diets and the influence of nutrient amendment of water on organism growth and sensitivity. Seven-day NaCl tests started with less than 24-h-old larvae were similar in sensitivity to 14-day and full life chronic tests, and were much more sensitive than those started with 7-day-old organisms. Reducing test temperature from 25 °C to 22 °C had a minor influence on culture timing, and little impact on sensitivity to NaCl. In other experiments, reconstituted test water supplemented with nutrients to potentially improve in-test food quality had minimal effect on growth at 7 days and did not significantly alter acute sensitivity to NaCl relative to unamended reconstituted water. A suitable commercially available, ready-to-feed diet substitute for cultured diatoms was not found. Testing N. triangulifer in effluents or receiving waters with the methods recommended will complement similar methods for Ceriodaphnia dubia. Environ Toxicol Chem 2022;41:2758-2768. © 2022 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Repellency of naturally occurring volatile alcohols to fungus gnat Bradysia sp. nr. coprophila (Diptera: Sciaridae) adults under laboratory conditions.

This study, conducted under laboratory conditions, was designed to determine the repellent activity of 10 naturally occurring volatile alcohol constituents against adults of the fungus gnat, Bradysia sp. nr. coprophila (Lintner) (Diptera: Sciaridae). The essential oil constituents were octanoic acid, furfural, acetophenone, benzaldehyde, dimethoxybenzene, borneol, menthol, 1-octen-3-ol, and 7-hydroxycitronellol, and alpha-terpineol. alpha-Terpineol, octanoic acid and furfural were tested at several concentrations, whereas the remaining seven were tested at only one concentration. The essential oil constituents' menthol, 1-octen-3-ol, and borneol displayed the most repellent activity. The mean percentage of fungus gnat adults recovered from the test compound petri dishes associated with the three essential oil constituents was between 6 and 15% compared with between 36 and 50% for the petri dishes with distilled water. The mean +/- SEM number of fungus gnat adults present in the sample compartments associated with menthol (10.4 +/- 2.6), 1-octen-3-ol (18.8 +/- 2.4), and borneol (23.4 +/- 5.6) was statistically lower than those in the petri dishes containing distilled water (60.9 +/- 7.4, 49.8 +/- 4.0, and 79.7 +/- 13.5), respectively. Only the highest concentration of alpha-terpineol (8.0 micromol) displayed significant repellent activity against fungus gnat adults. The other essential constituents tested, including octanoic acid (all three concentrations), furfural (both concentrations), acetophenone, dimethoxybenzene, and 7-hydroxycitronellol, were not statistically different from the distilled water control. The results of this study indicate that certain essential oil constituents repel fungus gnat adults, which may be useful, from a practical standpoint, in deterring adults from laying eggs into growing media.

Modeling the Bioavailability of Nickel and Zinc to Ceriodaphnia dubia and Neocloeon triangulifer in Toxicity Tests with Natural Waters.

We studied biotic ligand model (BLM) predictions of the toxicity of nickel (Ni) and zinc (Zn) in natural waters from Illinois and Minnesota, USA, which had combinations of pH, hardness, and dissolved organic carbon (DOC) more extreme than 99.7% of waters in a nationwide database. We conducted 7-day chronic tests with Ceriodaphnia dubia and 96-hour acute and 14-day chronic tests with Neocloeon triangulifer and estimated median lethal concentrations and 20% effect concentrations for both species. Toxicity of Ni and Zn to both species differed among test waters by factors from 8 (Zn tests with C. dubia) to 35 (Zn tests with N. triangulifer). For both species and metals, tests with Minnesota waters (low pH and hardness, high DOC) showed lower toxicity than Illinois waters (high pH and high hardness, low DOC). Recalibration of the Ni BLM to be more responsive to pH-related changes improved predictions of Ni toxicity, especially for C. dubia. For the Zn BLM, we compared several input data scenarios, which generally had minor effects on model performance scores (MPS). A scenario that included inputs of modeled dissolved inorganic carbon and measured Al and Fe(III) produced the highest MPS values for tests with both C. dubia and N. triangulifer. Overall, the BLM framework successfully modeled variation in toxicity for both Zn and Ni across wide ranges of water chemistry in tests with both standard and novel test organisms. Environ Toxicol Chem 2021;40:3049-3062. © 2021 SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Effects of millimeter wave carbon fibers on filter-feeding freshwater invertebrates.

The purpose of our study was to investigate the sub-lethal effects of millimeter wave carbon fibers (MWCF), a military obscurant, on filter-feeding freshwater invertebrates. We observed decreased survival, reproduction, and oxygen consumption in Daphnia magna at realistic loading rates. In experiments with the Asiatic clam (Corbicula fluminea), soft tissue dry weight and tissue condition index were not significantly different among control and MWCF exposed treatments; however, using a (15)N labeled alga as food, we observed decreased nitrogen turnover in tissues of clams exposed to MWCF, suggesting lower filtering or ingestion rates. Our findings combined with previous demonstrations of MWCF toxicity to green algae suggest that over a period of several months, bivalve growth may be inhibited, and cladoceran populations may be even more strongly affected by MWCF. Given that these fibers are persistent, further experiments should be conducted to determine the longer-term effects of contamination of water bodies with MWCF.

Acute and Chronic Toxicity of Nickel and Zinc to a Laboratory Cultured Mayfly (Neocloeon triangulifer) in Aqueous but Fed Exposures.

Aquatic insects are poorly represented in water quality criteria, and previous studies have suggested a lack of sensitivity in acute toxicity tests despite observational studies demonstrating the contrary. Our objectives were to determine the toxicity of nickel (Ni) and zinc (Zn) to the mayfly Neocloeon triangulifer in fed acute (96-h) and chronic exposures to estimate aqueous effect concentrations while acknowledging the importance of dietary exposure for these insects. For the chronic tests, we conducted preliminary full-life cycle (~25-30 d) and subchronic (14 d) exposures to compare the relative sensitivity of the 2 test durations under similar conditions (i.e., feeding rates). Observing similar sensitivity, we settled on 14 d as the definitive test duration. Furthermore, we conducted experiments to determine how much food could be added to a given volume of water while minimally impacting dissolved metal recovery; a ratio of food dry mass to water volume (<0.005) achieved this. In the 14-d tests, we obtained a median lethal concentration and most sensitive chronic endpoint of 147 and 23 µg/L dissolved Ni (acute to chronic ratio [ACR] = 6.4), respectively, and 81 (mean value) and 10 µg/L dissolved Zn (ACR = 8.1), respectively. The acute values are orders of magnitude lower than previously published values for mayflies, probably most importantly due to the presence of dietary exposure but also potentially with some influence of organism age and test temperature. Environ Toxicol Chem 2020;39:1196-1206. © 2020 SETAC.

Tonsillar surface swab bacterial culture results differ from those of the tonsillar core in recurrent tonsillitis.

OBJECTIVES/HYPOTHESIS: We aimed to determine whether there was a difference between core and surface bacteriology of Finnish adults with recurrent or chronic tonsillitis to understand whether a surface swab is worthwhile and which bacteria are involved. STUDY DESIGN: Case series. METHODS: Uninflamed tonsillar surface swabs and core biopsies were taken prior to and during surgery, respectively, in 103 patients aged 16 to 66 years undergoing tonsillectomy for recurrent or chronic tonsillitis. The McNemar test was used to determine differences between the surface and core in the most prevalent bacterial species. RESULTS: Twenty-seven bacterial species were isolated in addition to normal flora and were more commonly found in the core (1.11 surface and 4.75 core bacteria isolated per patient). Viridans group streptococci were the most commonly detected bacteria, found in 88% of the patients, mainly in the core. The bacteria in general were mainly isolated from the core. Of the 10 most prevalent bacteria, only group C ß-hemolytic streptococci showed no difference between detection from core and surface swabs. Other bacteria found mainly in the core include Prevotella melaninogenica, Staphylococcus aureus, and fusobacteria. CONCLUSIONS: There is discord between the surface and core bacteria. A different population of bacteria exists in the core, especially anaerobic bacteria, suggesting that a core sample may be useful in evaluating recurrent and chronic tonsillitis. LEVEL OF EVIDENCE: 4 Laryngoscope, 2019.

Mass spectrometry-based lipidomics of oral squamous cell carcinoma tissue reveals aberrant cholesterol and glycerophospholipid metabolism - A Pilot study.

Lipid metabolic reprogramming is one hallmark of cancer. Lipid metabolism is regulated by numerous enzymes, many of which are targeted by several drugs on the market. We aimed to characterize the lipid alterations in oral squamous cell carcinoma (OSCC) as a basis for understanding its lipid metabolism, thus identifying potential therapeutic targets. We compared lipid species, classes, and glycerophospholipid (GPL) fatty acid species between paired tumor tissue and healthy oral tongue mucosa samples from 10 OSCC patients using a QExactive mass spectrometer. After filtering the 1370 lipid species identified, we analyzed 349 species: 71 were significantly increased in OSCC. The GPL metabolism pathway was most represented by the lipids differing in OSCC (P = .005). Cholesterol and the GPLs phosphatidylcholines, phosphatidylethanolamines, and phosphatidylinositols were most significantly increased in OSCC tissue (FC 1.8, 2.0, 2.1, and 2.3 and, P = .003, P = .005, P = .002, P = .007). In conclusion, we have demonstrated a shift in the lipid metabolism in these OSCC samples by characterizing the detailed landscape. Predominantly, cholesterol and GPL metabolism were altered, suggesting that interactions with sterol regulatory binding proteins may be involved. The FA composition changes of the GPLs suggest increased de novo lipogenesis.

Factors Affecting Thermal Insulation of Songbird Nests as Measured Using Temperature Loggers.

AbstractBird nests represent an extended phenotype that is variable among and within species in terms of nest location and construction materials. It has also been suggested that nests indicate niche construction, although empirical evidence to support this is lacking. The nest wall is often considered to confer some insulation because this would help minimize the energetic expenditure by the incubating adult. However, it has been previously suggested that nests are constructed primarily for their structural role rather than for insulation, but to date, studies have used a variety of techniques to investigate the insulation of nest walls but only for relatively few species. This study used temperature loggers to determine insulatory values of nests for 16 previously undescribed species and to replicate values for eight previously described species. In addition, data for nest wall insulation that were determined using temperature loggers have been collated for a total of 32 different passerine species. The effects of nest dimensions, mass, and composition on these values were examined. Base thickness, but not wall thickness, and nest mass significantly positively affected insulatory values. This study found that the proportions of feathers and moss in the nest wall significantly positively correlated with insulatory values. This suggests that there is a key role for nest materials in determining insulation, which provides empirical evidence that nest building constitutes niche construction. The data will also help us interpret data from future studies of the thermal properties of nests. Ultimately, we need to develop our understanding of the role of insulatory values in the evolution of nest function.

Acute and Chronic Toxicity of Sodium Nitrate and Sodium Sulfate to Several Freshwater Organisms in Water-Only Exposures.

Elevated nitrate (NO3 ) and sulfate (SO4 ) in surface water are of global concern, and studies are needed to generate toxicity data to develop environmental guideline values for NO3 and SO4 . The present study was designed to fill existing gaps in toxicity databases by determining the acute and/or chronic toxicity of NO3 (tested as NaNO3 ) to a unionid mussel (Lampsilis siliquoidea), a midge (Chironomus dilutus), a fish (rainbow trout, Oncorhynchus mykiss), and 2 amphibians (Hyla versicolor and Lithobates sylvaticus), and to determine the acute and/or chronic toxicity of SO4 (tested as Na2 SO4 ) to 2 unionid mussels (L. siliquoidea and Villosa iris), an amphipod (Hyalella azteca), and 2 fish species (fathead minnow, Pimephales promelas and O. mykiss). Among the different test species, acute NO3 median effect concentrations (EC50s) ranged from 189 to >883 mg NO3 -N/L, and chronic NO3 20% effect concentrations (EC20s) based on the most sensitive endpoint ranged from 9.6 to 47 mg NO3 -N/L. The midge was the most sensitive species, and the trout was the least sensitive species in both acute and chronic NO3 exposures. Acute SO4 EC50s for the 2 mussel species (2071 and 2064 mg SO4 /L) were similar to the EC50 for the amphipod (2689 mg SO4 /L), whereas chronic EC20s for the 2 mussels (438 and 384 mg SO4 /L) were >2-fold lower than the EC20 of the amphipod (1111 mg SO4 /L), indicating the high sensitivity of mussels in chronic SO4 exposures. However, the fathead minnow, with an EC20 of 374 mg SO4 /L, was the most sensitive species in chronic SO4 exposures whereas the rainbow trout was the least sensitive species (EC20 > 3240 mg SO4 /L). The high sensitivity of fathead minnow was consistent with the finding in a previous chronic Na2 SO4 study. However, the EC20 values from the present study conducted in test water containing a higher potassium concentration (3 mg K/L) were >2-fold greater than those in the previous study at a lower potassium concentration (1 mg K/L), which confirmed the influence of potassium on chronic Na2 SO4 toxicity to the minnow. Environ Toxicol Chem 2020;39:1071-1085. © 2020 SETAC.

Comparing serum protein levels can aid in differentiating HPV-negative and -positive oropharyngeal squamous cell carcinoma patients.

BACKGROUND: The surrogate immunohistochemical marker, p16INK4a, is used in clinical practice to determine the high-risk human papillomavirus (HPV) status of oropharyngeal squamous cell carcinomas (OPSCC). With a specificity of 83%, this will misclassify some patients compared with direct HPV testing. Patients who are p16INK4a-positive but HPV DNA-negative, or RNA-negative, may be unsuitable for treatment de-escalation aimed at reducing treatment-related side effects. We aimed to identify cost-effective serum markers to improve decision making for patients at risk of misclassification by p16INK4a alone. METHODS: Serum proteins from pre-treatment samples of 36 patients with OPSCC were identified and quantified using label-free mass spectrometry-based proteomics. HPV-status was determined using p16INK4a/HPV DNA and E6/E7 mRNA. Serum protein expressions were compared between groups of patients according to HPV status, using the unpaired t-test with a Benjamini-Hochberg correction. ROC curves (AUC) were calculated with SPSS (v25). RESULTS: Of 174 serum proteins identified, complement component C7 (C7), apolipoprotein F (ApoF) and galectin-3-Binding Protein (LGALS3BP) significantly differed between HPV-positive and -negative tumors (AUC ranging from 0.84-0.87). ApoF levels were more than twice as high in the E6/E7 mRNA HPV-positive group than HPV-negative. CONCLUSIONS: Serum C7, ApoF and LGALS3BP levels discriminate between HPV-positive and HPV-negative OPSCC. Further studies are needed to validate these host immunity-related proteins as markers for HPV-associated OPSCC.

Influence of early neonatal experience on nociceptive responses and analgesic effects in rats.

Early maternal separation has profound effects on nociception in rats. Cross-fostering is a standard husbandry procedure used by some commercial breeders. This study aimed to determine if cross-fostering altered nociception and the analgesic efficacy of buprenorphine and morphine. At seven and nine weeks of age, an elevated plus maze was used to assess anxiety and Hargreaves apparatus was used to measure thermal nociception at two intensities in cross-fostered and naturally-reared rats. At 10 weeks of age these rats were assigned to one of three treatment groups: saline, buprenorphine or morphine. The Hargreaves apparatus was used to evaluate the effect of analgesics on nociception. Differences were observed in nociception between the cross-fostered and naturally-reared rats at both intensities. At the lower intensity no significant differences were seen between the cross-fostered and naturally-reared rats post-administration of an analgesic. At the higher intensity significant differences were apparent. Morphine was less effective in inducing analgesia to thermal stimuli in cross-fostered rats compared with naturally-reared rats, whereas the opposite was found with buprenorphine which had a more pronounced analgesic effect in the cross-fostered rats. No significant differences in performance on an elevated plus maze were demonstrated between the cross-fostered and naturally-reared rats.

The analgesic effects of oral paracetamol in two strains of mice undergoing vasectomy.

As the production of transgenic mice increases, the need for vasectomized mice also increases. Currently, there is no accurate method of identifying pain in these mice which can be used routinely and therefore no acceptable analgesic regimens can be established. Sixteen male CBA/CaCrl and 16 male DBA/2JCrl mice were randomly allocated to one of four treatment groups (saline, low, medium and high dose oral paracetamol) and then underwent abdominal vasectomy. Their behaviour was videotaped (filmed) preoperatively and at one hour postoperatively and the data were analysed using an automated system - HomeCageScan. HomeCageScan detected significant changes in 16 behaviours following vasectomy. Such behaviours included twitching, rearing and grooming with varying levels of significance between the strains. No significant effects of drug treatment in any of the behaviours analysed by HomeCageScan were detected in the postoperative observation period. Strain-specific changes do occur in behaviour following abdominal vasectomy and HomeCageScan is capable of detecting these changes. The use of orally administered paracetamol was deemed to be an ineffective analgesic for CBA/CaCrl and DBA/2JCrl mice undergoing vasectomy.