Human papillomavirus 16 infection has been proven to be associated with oropharyngeal squamous cell carcinomas (SCCs) and is probably the main reason of the reported increase in the incidence. The ...role of high-risk (HR) HPV for carcinogenesis of other sites in the head and neck awaits confirmation. With the aim to evaluate the prevalence of HPV infection and the reliability of different diagnostic tools in SCCs of different sites, 109 consecutive untreated head and neck SCCs were enrolled, and fresh tumour samples collected.
Human papillomavirus DNA was detected by Digene Hybrid Capture 2 (HC2). Human papillomavirus E6 and E7 mRNA were detected by NucliSENS EasyQ HPVv1. P16 expression was evaluated by immunohistochemistry.
In all, 12.84% of cases were infected by HR genotypes and 1.84% by low-risk genotypes. Human papillomavirus 16 accounted for 87% of HR infections. The overall agreement between DNA and RNA detection is 99.1%. Although p16 expression clearly correlates with HPV infection (P=0.0051), the inter-rater agreement is poor (k=0.27). The oropharynx showed the highest HR HPV infection rate (47.6%) and was also the only site in which p16 immunohistochemistry revealed to be a fair, but not excellent, diagnostic assay (κ=0.61).
The prognostic role of HR HPV infection in oropharyngeal oncology, with its potential clinical applications, underscores the need for a consensus on the most appropriate detection methods. The present results suggest that viral mRNA detection could be the standard for fresh samples, whereas DNA detection could be routinely used in formalin-fixed, paraffin-embedded samples.
Overexpression of the c-Met/hepatocyte growth factor receptor(HGF-R) proto-oncogene and abnormal generation of intracellular oxygen species (reactive oxygen species (ROS)) have been linked, by ...independent lines of evidence, to cell transformation and to malignant growth. By comparing two subpopulations of the B16 mouse melanoma (B16-F0 and B16-F10) endowed with different lung metastasis capacities (low and high, respectively) we found that both the expression/phosphorylation of c-Met and the steady-state levels of ROS positively correlated with metastatic growth. shRNA-mediated downregulation of c-Met in F10 cells led to a parallel decrease in the generation of oxygen species and in metastatic capacity, suggesting that oxidants may mediate the pro-metastatic activity of the HGF receptor. c-Met activation by a ligand elicits the formation of oxidant species through the oxidase-coupled small GTPase Rac-1, a relevant downstream target of the HGF-R. Moreover, cell treatment with the catalytic ROS scavengers EUK-134 and EUK-189 attenuates Met signaling to ERKs and inhibits the anchorage-independent growth of F10 cells, consistent with a critical role for oxygen species in HGF signaling and in aggressive cell behavior. Finally, genetic manipulation of the Rac-ROS cascade at different levels demonstrated its crucial role in the pro-metastatic activity of c-Met in vivo. Thus, we have outlined a novel cascade triggered by c-Met and mediated by ROS, linked to metastasis and potentially targetable by new antimetastatic, redox-based therapies.
Even though premating isolation is hypothesized to be a major driving force in speciation, its genetic basis is poorly known. In the noctuid moth Heliothis subflexa, one group of sex pheromone ...components, the acetates, emitted by the female, plays a crucial isolating role in preventing interspecific matings to males of the closely related Heliothis virescens, in which females do not produce acetates and males are repelled by them. We previously found intraspecific variation in acetates in H. subflexa: females in eastern North America contain significantly more acetates than females in Western Mexico. Here we describe the persistence of this intraspecific variation in laboratory‐reared strains and the identification of one major quantitative trait locus (QTL), explaining 40% of the variance in acetate amounts. We homologized this intraspecific QTL to our previously identified interspecific QTL using restriction‐associated DNA (RAD) tags. We found that a major intraspecific QTL overlaps with one of the two major interspecific QTL. To identify candidate genes underlying the acetate variation, we investigated a number of gene families with known or suspected acetyl‐ or acyltransferase activity. The most likely candidate genes did not map to our QTL, so that we currently hypothesize that a transcription factor underlies this QTL. Finding a single, large QTL that impacts variation in pheromone blends between and within species is, to our knowledge, the first such example for traits that have been demonstrated to affect premating isolation.
The article documents faculty experiences with the shift online due to the pandemic and provides recommendations to science, technology, engineering, and mathematics (STEM) instructors. Over 100 ...faculty members were surveyed on these topics and contrasted with previously reported student experiences. The online shift changed how faculty administered exams, ran courses, and acted to ensure academic integrity. For example, when exams went online, 73% of faculty reported spending more time preventing cheating. Concerning academic integrity and stress, faculty and students agreed with the exception of a few notable disconnects. Students reported greater workloads in online classes, while faculty maintained that the shift online did not change student workloads. Students perceived more online cheating than faculty. Overall, there seems to be a significant disconnect regarding faculty not realizing how much their actions may encourage or discourage cheating. Few faculty (<15%) indicated that being a tough grader or having test times too short is a motivating factor, but over 55% of students reported that these motivate students to cheat. Conversely, over 60% of students reported respect for their professors discourages them from cheating, while only 37% of faculty indicated the same. Over 70% of faculty and students indicated that fear of getting caught is a deterrent to cheating. Recommendations to reconnect include (i) faculty should use the finding that the number one deterrent of cheating is fear of getting caught; and (ii) faculty should maintain students' respect by being clear or overestimating workload requirements, carefully adjusting time for online exams, and setting clear expectations with uncomplicated exam questions consistent with the material taught.
Objective Full-field visual evoked potentials (ff-VEP) are fundamental in neurological practice to define the nature of visual disturbances. We explored whether, in case of normal ff-VEP and ...suspected organic visual pathway involvement, multifocal visual evoked potentials (mf-VEP) and optical coherence tomography (OCT) can be useful in the diagnostic workup. Methods Observational case reports. Results Three patients arrived at our department for visual disturbances. Two had relapsing optic neuritis (ON) and visual acuity (VA) loss, with normal or non-significant ff-VEP outside acute clinical episodes. In both cases OCT showed a retinal nerve fiber layer (RNFL) thickness reduction, allowing visual pathway damage identification. The third, with meningioma involving left optic nerve, complained of blurred vision in the nasal field of the left eye as confirmed by computerized perimetry (CP). While ff-VEP showed normal latency and non-significant amplitude reduction in the left eye, mf-VEP showed important amplitude reduction in the whole lower left eye field. OCT scan confirmed axonal damage showing left RNFL thinning. Conclusions Sometimes ff-VEP fails in identifying abnormalities in patients with processes involving the visual pathway, particularly in cases with axonal or sectoral optic nerve involvement. We suggest the importance of a multimodal evaluation, including OCT and mf-VEP.
Cytomegalovirus (CMV) disease represents a major cause of post-transplantation morbidity and mortality. To estimate the risk of infection and monitor response to antiviral therapy, current guidelines ...suggest combination of viral load monitoring with direct assessment of CMV-specific immune response. We used enzyme-linked immunospot (ELISpot) for the evaluation of CMV-specific T-cell response in kidney transplant recipients with CMV viremia and investigated how information gained could help manage CMV infection.
Seventeen patients on pre-emptive antiviral therapy and CMV quantitative polymerase chain reaction (qPCR) ≥500 copies/mL (first episode after transplantation) were assessed using ELISpot and divided into Weak (9 patients with baseline ELISpot <25 spot-forming colonies SFCs/200,000 peripheral blood mononuclear cells PBMCs) and Strong Responders (8 patients with baseline ELISpot ≥25 SFCs/200,000 PBMCs). CMV-specific T-cell response, infection severity, viral load, and antiviral therapy were prospectively recorded and compared between groups at 1, 2, and 24 months of follow-up.
Demographic and transplant characteristics of Weak and Strong Responders were similar. No episodes of CMV disease were observed. Weak Responders were more likely to experience CMV syndrome (56% vs 36.5%) and late virus reactivation (56% vs 25%) than Strong Responders. Weak Responders showed higher baseline median viral loads (19,700 vs 9265 copies/mL) and needed antiviral therapy for longer (179 vs 59.5 days). T-cell response showed 2 main patterns: early and delayed.
ELISpot provides prognostic information about infection severity, risk of late reactivation, and response to therapy. Randomized trials, evaluating the need for antiviral therapy in kidney transplant recipients with asymptomatic infection and effective virus-specific T-cell immune response, are warranted.
•ELISpot shows CMV-specific T-cell response in kidney transplant recipients.•CMV-specific T-cell response influences clinical course of the disease.•Strong virus-specific immune response can mitigate CMV infection.•Antiviral therapy could be tailored according to ELISpot results.
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