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Abstract Background Psychosocial factors and socioeconomic status have been associated with incidence, survival, and quality of life among patients with head and neck cancer. We investigated the association between different psychosocial factors, socioeconomic status, and patient delays in T3–T4 oral, oropharyngeal, and laryngeal cancer. Patients and methods We conducted a nationwide prospective questionnaire-based study (n = 203) over a 3-year period. Results We found no association between psychosocial factors (depression, social isolation, loneliness, and cynical hostility) and patient delay. Depression was three times more common among head and neck cancer patients compared with the general Finnish population. Head and neck cancer patients had lower educational levels and employment status, and were more often current smokers and heavy drinkers. Conclusions Although we found no association between patient delay and psychosocial factors, patients diagnosed with a large head and neck cancer appeared to have a lower socioeconomic status and higher risk for developing depression, which should be considered in clinical practice.

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Plant-based protein is considered a sustainable protein source and has increased in demand recently. However, products containing plant-based proteins require further modification to achieve the desired functionalities akin to those present in animal protein products. This study aimed to investigate the effects of enzymes as cross-linking reagents on the physicochemical and functional properties of hybrid plant- and animal-based proteins in which lupin and whey proteins were chosen as representatives, respectively. They were hybridised through enzymatic cross-linking using two laccases (laccase R, derived from Rhus vernicifera and laccase T, derived from Trametes versicolor) and transglutaminase (TG). The cross-linking experiments were conducted by mixing aqueous solutions of lupin flour and whey protein concentrate powder in a ratio of 1:1 of protein content under the conditions of pH 7, 40 °C for 20 h and in the presence of laccase T, laccase R, or TG. The cross-linked mixtures were freeze-dried, and the powders obtained were assessed for their cross-linking pattern, colour, charge distribution (ζ-potential), particle size, thermal stability, morphology, solubility, foaming and emulsifying properties, and total amino acid content. The findings showed that cross-linking with laccase R significantly improved the protein solubility, emulsion stability and foaming ability of the mixture, whereas these functionalities were lower in the TG-treated mixture due to extensive cross-linking. Furthermore, the mixture treated with laccase T turned brownish in colour and showed a decrease in total amino acid content which could be due to the enzyme’s oxidative cross-linking mechanism. Also, the occurrence of cross-linking in the lupin and whey mixture was indicated by changes in other investigated parameters such as particle size, ζ-potential, etc., as compared to the control samples. The obtained results suggested that enzymatic cross-linking, depending on the type of enzyme used, could impact the physicochemical and functional properties of hybrid plant- and animal-based proteins, potentially influencing their applications in food.

Faba bean is a promising source of ingredients for the production of meat analogs. However, sensory properties of faba bean, especially the bitter taste of the protein concentrate, restrict its use. Our aim was to assess the feasibility of two types of faba bean ingredients—flour (from germinated, gently heat-treated beans) and groat (from non-germinated, roasted beans)—in combination with pea protein isolate and oat fiber concentrate for producing meat analogs using high-moisture extrusion. We produced six samples using varying recipes, while maintaining constant process parameters. An untrained panel (55 participants) evaluated the samples for key sensory attributes (check-all-that-apply) and rated their pleasantness. The water absorption capacity and mechanical properties of the samples were assessed using instrumental measurements. The samples were frequently described as ‘beany’ and ‘tasteless’, but very rarely as ‘bitter’. The most frequently cited attributes for mouthfeel varied between the samples containing 30% (‘tough’, ‘gummy’) and 50% (‘crumbly’, ‘floury’) of faba bean flour/groat and were associated with corresponding mechanical properties. On average, the sample containing a blend of faba bean groat and pea protein isolate (50% each) appeared to be the most pleasant. Our results suggest that faba bean groat with pea protein isolate enables the production of fibrous meat analogs with acceptable taste and texture, without the bitter off-taste.

Lupin is a nutritious, yet undervalued grain used as a fodder and food crop. In the present study, native lupin flour (LF), lupin protein concentrate (LPC), and lupin protein isolate (LPI) were combined (70% LPI:LPC blend ratios [30:70, 50:50, and 70:30] and 30% LF constant fraction), extruded at high moisture (45–55%), and shaped with a long cooling die (800 mm) to obtain texturized meat analogues (TMAs) with fibrous structures. The characteristics of TMAs (e.g., hardness, water hydration capacity) depended heavily on water content, blend ratios (LPI:LPC), and to a lesser extent, the long cooling die temperature. Color changes (i.e., L*, b*) were mostly attributed to variations in blend ratios (LPI:LPC). Microstructure analysis showed that TMAs with higher water content (55%) were more likely to have thinner walls and smaller void thickness. Fluorescence imagery revealed that TMAs with lower LPI content presented more homogeneous structures. These findings show that reasonable amounts (30% d.m.) of native lupin flour can be incorporated into meat analogues by maintaining a sufficiently high protein content (>50% d.m.) to trigger the formation of fibrous structures.

Legumes have been recognised as healthy and environmentally friendly protein sources. Knowledge about the vitamin B contents in legume ingredients and extrudates is scarce. In this study, we investigated thiamin, riboflavin, niacin, and folate in various faba bean, lupin, and pea ingredients. Further, the retention of B vitamins in high moisture extrusion was studied. Prior to liquid chromatographic determinations of thiamin, riboflavin, niacin, and folate, vitamins were extracted by acid hydrolysis (niacin), enzymatic treatment (folate), or their combination (thiamin and riboflavin). The contents (on a dry matter basis) varied greatly among different ingredients: the thiamin content was 0.2–14.2 µg/g; riboflavin, 0.3–5.9 µg/g; niacin, 8.8–35.5 µg/g, and folate, 45–1453 ng/g. Generally, the highest levels were in flours and protein concentrates, whereas low levels were observed in isolates. The retention of B vitamins was excellent in high moisture extrusion, except for folate in faba bean, where the folate contents were 42–67% lower in the extrudates than in the respective ingredient mixtures. In terms of both vitamin B contents and their retention, extrudates containing substantial amounts of flour or protein concentrate are promising plant-based sources of thiamin, riboflavin, niacin, and folate.

ABSTRACT Enteroviruses are a group of positive single-stranded viruses that belong to the Picornaviridae family. They regularly infect humans and cause symptoms ranging from the common cold and hand-foot-and-mouth disease to life-threatening conditions, such as dilated cardiomyopathy and poliomyelitis. Enteroviruses have also been associated with chronic immune-mediated diseases, such as type 1 diabetes, celiac disease, and asthma. Studying these disease-pathogen connections is challenging due to the high prevalence of enterovirus infections in the population and the transient appearance of the virus during the acute infection phase, which limit the identification of the causative agent via methods based on the virus genome. Serological assays can detect the antibodies induced by acute and past infections, which is useful when direct virus detection is not possible. We describe in this immuno-epidemiological study how the antibody levels against VP1 proteins from eight different enterovirus types, representing all seven of the human infecting enterovirus species, vary over time. VP1 responses first significantly (P < 0.001) decline until 6 months of age, reflecting maternal antibodies, and they then start to increase as the infections accumulate and the immune system develops. All 58 children in this study were selected from the DiabImmnune cohort for having PCR-confirmed enterovirus infections. Additionally, we show that there is great, although not complete, cross-reactivity of VP1 proteins from different enteroviruses and that the response against 3C-pro could reasonably well reflect the recent Enterovirus infection history (ρ = 0.94, P = 0.017). The serological analysis of enterovirus antibodies in sera from children paves the way for the development of tools for monitoring the Enterovirus epidemics and associated diseases. IMPORTANCE Enteroviruses cause a wide variety of symptoms ranging from a mild rash and the common cold to paralyzing poliomyelitis. While enteroviruses are among the most common human pathogens, there is a need for new, affordable serological assays with which to study pathogen-disease connections in large cohorts, as enteroviruses have been linked to several chronic illnesses, such as type 1 diabetes mellitus and asthma exacerbations. However, proving causality remains an issue. In this study, we describe the use of an easily customizable multiplexed assay that is based on structural and nonstructural enterovirus proteins to study antibody responses in a cohort of 58 children from birth to 3 years of age. We demonstrate how declining maternal antibody levels can obscure the serological detection of enteroviruses before the age of six months and how antibody responses to nonstructural enterovirus proteins could be interesting targets for serodiagnosis.

Oat has been recognized for its health-promoting fiber, β-glucan, while protein-rich faba bean has remained underutilized in Nordic countries despite its good nutritional quality. This research investigated the functionality of oat fiber concentrate and faba bean protein concentrate in plant-based substitutes for minced meat (SMs). The resulting product aimed at mimicking the mechanical and physicochemical characteristics of beef minced meat (BM) and its applications (i.e., fried and burger patty). In this regard, the mechanical properties (e.g., chewiness, Young's modulus) of original/fried SMs were comparable to or higher than those of original/fried BM. SM patties (45% SMs) were structurally weaker than beef burger patties (100% BM). The rheological analysis showed that the presence of oat fiber concentrate increased the gel-like properties of the blend, which correlated with the overall strength of original SMs (e.g., Young's modulus). The results suggested that SMs could be used as BM for the preparation of vegetarian meat-like products.

Cells shape their extracellular milieu by secreting intracellular products into the environment including extracellular vesicles which are lipid-bilayer limited membrane particles. These vesicles carry out a range of functions, including regulation of coagulation, via multiple contributor mechanisms. Urinary extracellular vesicles are secreted by various cells, lining the urinary space, including the nephron and bladder. They are known to have procoagulant properties, however, the details of this function, beyond tissue factor are not well known. The aim of the study was to access the role of urinary extracellular vesicles in impacting coagulation upon supplementation to plasma. This could indicate their physiological function upon kidney injury or pathology. Supplementation to standard human plasma and plasmas deficient in various coagulation factors was used for this purpose, and calibrated automated thrombogram (CAT®) was the major technique applied. We found that these vesicles contain multiple coagulation-related factors, and their lipid composition affects coagulation activities of plasma upon supplementation. Remarkably, these vesicles can restore thrombin generation in FVII, FVIII, FIX and FXI -deficient plasmas. This study explores the multiple roles of urinary extracellular vesicles in coagulation in in vitro blood coagulation and implies their importance in its regulation by several mechanisms.

The bioaccessibility of vitamin B12 (B12) in plant-based products fortified using wheat bran extract fermented with B12-producing food-grade Propionibacterium freudenreichii was studied by applying a standard static in vitro model. At first, a culture of P. freudenreichii, fresh or heat-treated, was subjected to in vitro assays. Then, food ingredients or products were evaluated for their in vitro bioaccessibility: spray-dried wheat bran extract powder, pasta made with an extruder using fermented bran extract and breads made with spray-dried powder or with added cyanocobalamin. B12 bioaccessibility from the fresh P. freudenreichii culture was only ca. 53%, which, when heated, increased to 73%. The bioaccessibility of B12 from the food products varied from 75% (spray-dried powder) to 95% (breads). B12 from the fortified bread was as bioaccessible as from the bread made with added cyanocobalamin (99%). The in vitro results suggest that B12 synthesized by P. freudenreichii, when fortified in the studied cereal-based products, is largely bioaccessible and could be available for absorption. Plant-based products fortified using fermentation with P. freudenreichii could thus be considered excellent sources of bioaccessible B12.

In type 3 von Willebrand disease (VWD3), the most severe form with absent von Willebrand factor (VWF), the bleeding phenotype is variable. Platelet contribution to the hemostatic defect in VWD3 calls upon further studies. We investigated the contribution of platelets to in vitro thrombin generation (TG) and platelet procoagulant activity in VWD3. TG was assessed by calibrated automated thrombogram (CAT) in platelet-poor (PPP) and –rich plasma (PRP) from 9 patients before and in 6 patients also 30 min after receiving their regular VWF therapy. Responsiveness of PPP to FVIII and protein S was also investigated. TG data were compared with routine laboratory variables, rotational thromboelastometry (ROTEM) and platelet expression of P-selectin and phosphatidylserine in flow cytometry. Compared with healthy controls, TG was markedly decreased in VWD3 PPP (peak thrombin was 16% of normal median), but not in PRP (77% of normal median) (p = 0.002). Six out of nine patients (67%) were high responders in their platelet P-selectin, and 5/9 (56%) in phosphatidylserine expression. Replacement therapy improved TG in PPP, while in PRP TG only modestly increased or was unaffected. In PPP, FVIII levels associated with TG and in vitro FVIII-supplemented TG inclined up to threefold. Conversely, a FVIII inhibitory antibody reduced plasma TG in all, but especially in patients with remnant FVIII levels. Inhibition of protein S improved plasma TG, particularly at low FVIII levels. ROTEM failed to detect VWD3. In VWD3, TG is reduced in PPP and regulated by FVIII and protein S, but TG is close to normal in PRP. VWD3 platelets seem to compensate for the FVIII-associated reduction in TG by their exposure of P-selectin and phosphatidylserine.

Hemostasis, thrombosis, and inflammation are tightly interconnected processes which may give rise to thrombo-inflammation, involved in infectious and non-infectious acute and chronic diseases, including cardiovascular diseases (CVD). Traditionally, due to its hemostatic role, blood coagulation is isolated from the inflammation, and its critical contribution in the progressing CVD is underrated, until the full occlusion of a critical vessel occurs. Underlying vascular injury exposes extracellular matrix to deposit platelets and inflammatory cells. Platelets being key effector cells, bridge all the three key processes (hemostasis, thrombosis, and inflammation) associated with thrombo-inflammation. Under physiological conditions, platelets remain in an inert state despite the proximity to the endothelium and other cells which are decorated with glycosaminoglycan (GAG)-rich glycocalyx (GAGs). A pathological insult to the endothelium results in an imbalanced blood coagulation system hallmarked by increased thrombin generation due to losses of anticoagulant and cytoprotective mechanisms, i.e., the endothelial GAGs enhancing antithrombin, tissue factor pathway-inhibitor (TFPI) and thrombomodulin-protein C system. Moreover, the loss of GAGs promotes the release of mediators, such as von Willebrand factor (VWF), platelet factor 4 (PF4), and P-selectin, both locally on vascular surfaces and to circulation, further enhancing the adhesion of platelets to the affected sites. Platelet-neutrophil interaction and formation of neutrophil extracellular traps foster thrombo-inflammatory mechanisms exacerbating the cardiovascular disease course. Therefore, therapies which not only target the clotting mechanisms but simultaneously or independently convey potent cytoprotective effects hemming the inflammatory mechanisms are expected to provide clinical benefits. In this regard, we review the cytoprotective protease activated protein C (aPC) and its strong anti-inflammatory effects thereby preventing the ensuing thrombotic complications in CVD. Furthermore, restoring GAG-like vasculo-protection, such as providing heparin-proteoglycan mimetics to improve regulation of platelet and coagulation activity and to suppress of endothelial perturbance and leukocyte-derived pro-inflammatory cytokines, may provide a path to alleviate thrombo-inflammatory disorders in the future. The vascular tissue-modeled heparin proteoglycan mimic, antiplatelet and anticoagulant compound (APAC), dual antiplatelet and anticoagulant, is an injury-targeting and locally acting arterial antithrombotic which downplays collagen- and thrombin-induced and complement-induced activation and protects from organ injury.

Faba bean is a potential ingredient due to its high protein yield and its possible cultivation in colder climate regions. In this study, meat analogues made from faba bean protein isolate (FPI) and concentrate (FPC) blends were produced using high moisture extrusion. The aim of this study was to investigate the effect of the FPI content (FPIc), feed water content (FWC), and temperature of the long cooling die (LT) during extrusion on the mechanical and physicochemical properties as well as on the structure of the meat analogues. Increased FPIc resulted in higher values in hardness, gumminess, chewiness, and cutting strengths as well as in darker colour and decreased water absorption capacity. The effect of increased FWC on these properties was weaker and the opposite. Images from microtomography revealed that higher FPIc led to a less organised fibrous structure. In conclusion, fibrous structures can be achieved by utilising a mixture of faba bean protein ingredients, and a higher FPC content seemed to promote fibre formation in the meat analogue.

Millennials are considered the key generation with regard to the consumption of plant-based meat alternatives via flexitarianism. This study sought to characterize millennials’ consumer segments based on their consumption of and attitudes toward meat and meat alternatives. We conducted an online survey on the hedonic tones of the associations evoked by meat and meat alternatives, consumption of such foods, and diet-related attitudes among a representative sample of Finnish millennials (N = 546, 59% women, age 20–39 years). Some 41% of respondents regularly ate plant-based meat alternatives, while 43% had tried such foods. We divided the respondents into six segments based on the hedonic tones of their meat vs. meat alternatives associations. The segments differed in terms of their consumption of meat alternatives and the underlying reasons why, importance of meat in meals, and Meat Commitment Scale scores. The segment that reported much more positive associations with meat than meat alternatives (~14% of the respondents) may prove resistant to interventions intended to reduce meat intake, whereas the segment that displayed the most positive attitudes toward meat alternatives (~18%) did not eat much meat. Thus, the four middle segments (totaling ~68%), whose associations’ hedonic tones were close to each other, may be the best targets for future interventions designed to reduce meat consumption through the use of meat alternatives. To conclude, introducing a simple segmentation allowed us to identify consumer segments with large potential to reduce meat consumption.

The immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptor G6b-B is critical for platelet production and activation. Loss of G6b-B results in severe macrothrombocytopenia, myelofibrosis and aberrant platelet function in mice and humans. Using a combination of immunohistochemistry, affinity chromatography and proteomics, we identified the extracellular matrix heparan sulfate (HS) proteoglycan perlecan as a G6b-B binding partner. Subsequent in vitro biochemical studies and a cell-based genetic screen demonstrated that the interaction is specifically mediated by the HS chains of perlecan. Biophysical analysis revealed that heparin forms a high-affinity complex with G6b-B and mediates dimerization. Using platelets from humans and genetically modified mice, we demonstrate that binding of G6b-B to HS and multivalent heparin inhibits platelet and megakaryocyte function by inducing downstream signaling via the tyrosine phosphatases Shp1 and Shp2. Our findings provide novel insights into how G6b-B is regulated and contribute to our understanding of the interaction of megakaryocytes and platelets with glycans.

Bone metastases are prevalent in many common cancers such as breast, prostate, and lung cancers, and novel therapies for treating bone metastases are needed. Human immune system-engrafted models are used in immuno-oncology (IO) studies for subcutaneous cancer cell or patient-derived xenograft implantations that mimic primary tumor growth. Novel efficacy models for IO compounds on bone metastases need to be established. The study was performed using CIEA NOG (NOG) mice engrafted with human CD34+ hematopoietic stem cells (huNOG) and age-matched immunodeficient NOG mice. Bone phenotyping was performed to evaluate baseline differences. BT-474 human breast cancer cells were inoculated into the tibia bone marrow, and cancer-induced bone changes were monitored by X-ray imaging. Bone content and volume were analyzed by dual X-ray absorptiometry and microcomputed tomography. Tumor-infiltrating lymphocytes (TILs) and the expression of immune checkpoint markers were analyzed by immunohistochemistry. Bone phenotyping showed no differences in bone architecture or volume of the healthy bones in huNOG and NOG mice, but the bone marrow fat was absent in huNOG mice. Fibrotic areas were observed in the bone marrow of some huNOG mice. BT-474 tumors induced osteoblastic bone growth. Bone lesions appeared earlier and were larger, and bone mineral density was higher in huNOG mice. huNOG mice had a high number of human CD3-, CD4-, and CD8-positive T cells and CD20-positive B cells in immune-related organs. A low number of TILs and PD-1-positive cells and low PD-L1 expression were observed in the BT-474 tumors at the endpoint. This study reports characterization of the first breast cancer bone growth model in huNOG mice. BT-474 tumors represent a “cold” tumor with a low number of TILs. This model can be used for evaluating the efficacy of combination treatments of IO therapies with immune-stimulatory compounds or therapeutic approaches on bone metastatic breast cancer.

Increased ultraviolet-B (UV-B) due to global change can affect plant development and metabolism. Quinoa tolerates extreme conditions including high UV levels. However, the physiological mechanisms behind its abiotic stress tolerance are unclear, especially those related to UV-B. We previously demonstrated that 9.12 kJ m−2 d−1 may induce UV-B-specific signaling while 18.24 kJ m−2 d−1 promotes a UV-B-independent response. Here, we explored the effects of these UV-B doses on hormonal regulation linked to plant morphology and defense among diverse varieties. Changes in fluorescence parameters of photosystem II, flavonoids and hormones (indoleacetic acid (IAA), jasmonic acid (JA), abscisic acid (ABA) and salicylic acid (SA)) were surveyed under controlled conditions. Here, we showed that the sensitivity to short acute UV-B doses in varieties from different habitats is influenced by their parental lines and breeding time. UV-B sensitivity does not necessarily correlate with quinoa’s geographical distribution. The role of flavonoids in the UV-B response seems to be different depending on varieties. Moreover, we found that the extent of changes in JA and SA correlate with UV-B tolerance, while the increase of ABA was mainly related to UV-B stress.

Quinoa epitomizes the drive for healthier foods with ethnic concepts in developed countries, particularly among millennials. As a result, the popularity of quinoa as a gluten-free alternative has steadily grown over the last 20 years. Despite this, little is known about the impact of specific varieties on processed foods. The purpose of this study was to examine the impact of quinoa varieties (variety and content) on the mechanical and textural properties of buckwheat-based extruded pasta (spaghetti). Peruvian native (var. rosada taraco, kuchivila, negra collana, and mistura) and Latvian-grown (var. titicaca) varieties were independently incorporated to pasta between 5 and 20% (w/w). Pasta containing 20% quinoa var. negra collana, which presented the largest content of fiber and lowest content of saponin, was strongly associated to structural resilience (i.e., cohesiveness, firmness). Conversely, pasta containing 20% quinoa var. Titicaca appeared structurally weak (i.e., smooth). The addition of saponin-containing varieties to pasta (20%), such as rosada taraco and mistura, resulted in resilient structures with little effect on taste (incl. bitterness). Despite initial stability, pasta containing 20% quinoa var. kuchivila suffered heavy structural damage. In conclusion, the relationship of compositional, mechanical, and textural properties of pasta was strongly variety-dependent.

Co-design is used for improving innovation, obtaining better solutions, and higher user satisfaction. In this paper we present how the use of a walk-in virtual environment and actual-size virtual prototypes support co-design. Unlike in most studies we presented the prototypes to users in an early phase of the design process. This study examines the co-design of healthcare facilities with multi-occupational groups. The practical case examines designing single-patient rooms for an intensive care unit. In this design process 238 participants of different hospital professions evaluated virtual prototypes in three iterative rounds. The participants improved the design by discussing their work practices. The virtual environment situation and actual size virtual prototypes make an easy environment for users to discuss and evaluate the design without any design knowledge. In addition to describing the co-design results we also outline some important issues and guidelines about creating the virtual prototypes and organizing the participants’ visits in a virtual environment.

The relationship between the efficacy of resuscitation and the mattresses and backboards used in acute care units, has been studied previously. However, few reports focus on the relative efficacy of resuscitation when using mattresses with different modes of function. This study examines the performance of different support surfaces during experimental cardiopulmonary resuscitation (CPR). The surfaces included a hard surface, a higher specification foam mattress, a dynamic, alternating pressure mattress, and a dynamic, reactive minimum pressure air mattress system. A pressure sensitive mat was placed between the mattresses and each surface and the efficacy of resuscitation measured using differences in compression frequency, compression depth and hands-on time. Our results suggest that the efficacy of resuscitation is dependent on the mode of action of the mattress, while adequate compression frequency and depth do not have a significant effect. In the open system alternating mattress, deflation of the mattress using the CPR function improved the stability of the resuscitation in our study, especially in situations where the height of the air mattress is greater than 20–25 centimeters. Using our experimental system, resuscitation on a closed air system mattress optimally combined stability and effort, while the CPR function converts the air system of the mattress to open, which impairs its functionality during resuscitation. These results indicate that resuscitation is dependent of the mode of action of the mattress and whether the mattress-specific CPR function was used or not. However, the interactions are complex and are dependent on the interaction between the body and the mattress, i.e. its immersion and envelopment properties. Furthermore, this study casts doubt on the necessity of the CPR function in air mattresses.

Abstract Background Epithelial-mesenchymal transition (EMT) is a crucial process in tumorigenesis since tumor cells attain fibroblast-like features enabling them to invade to surrounding tissue. Two transcription factors, TWIST and SNAI1, are fundamental in regulating EMT. Methods Immunohistochemistry was used to study the expression of TWIST and SNAI1 in 109 pharyngeal squamous cell carcinomas. Results Tumors with intense stromal staining of TWIST relapsed more frequently (p = 0.04). Tumors with both positive TWIST and SNAI1 immunoreactivity in the stroma were at least Stage II (p = 0.05) and located more often in hypopharynx (p = 0.035). Tumors with negative immunostaining of TWIST and SNAI1 in the stromal compartment were smaller (T1-2) (p = 0.008), less advanced (SI-II) (p = 0.031) and located more often in the oropharynx (p = 0.007). Patients with negative SNAI1 and TWIST immunostaining in tumor stroma had a better 5-year disease-specific and overall survival (p = 0.037 and p = 0.014 respectively). Conclusion TWIST and SNAI1 expression in stromal cells is associated with clinical and histopathological characteristics that indicate progressive disease. Negative expression of these EMT-promoting transcription factors predicts a better outcome.