Human CD4 cytotoxic T lymphocytes mediate potent tumor control in humanized immune system mice.

Efficacy of immune checkpoint inhibitors in cancers can be limited by CD8 T cell dysfunction or HLA-I down-regulation. Tumor control mechanisms independent of CD8/HLA-I axis would overcome these limitations. Here, we report potent CD4 T cell-mediated tumor regression and memory responses in humanized immune system (HIS) mice implanted with HT-29 colorectal tumors. The regressing tumors showed increased CD4 cytotoxic T lymphocyte (CTL) infiltration and enhanced tumor HLA-II expression compared to progressing tumors. The intratumoral CD4 T cell subset associated with tumor regression expressed multiple cytotoxic markers and exhibited clonal expansion. Notably, tumor control was abrogated by depletion of CD4 but not CD8 T cells. CD4 T cells derived from tumor-regressing mice exhibited HLA-II-dependent and tumor-specific killing ex vivo. Taken together, our study demonstrates a critical role of human CD4 CTLs in mediating tumor clearance independent of CD8 T cells and provides a platform to study human anti-tumor immunity in vivo.

Attitudes of Young Tri-City Residents toward Game Meat in the Context of Food Neophobia and a Tendency to Look for Diversity in Food.

To conduct rational hunting management, a certain number of wild animals must be harvested yearly. However, some countries have a problem with managing the harvested meat. An example is Poland, where game consumption is estimated at 0.08 kg/person/year. This situation leads to environmental pollution as a result of meat exports. The level of environmental pollution depends on the type of transport and distance. However, the use of meat in the country of harvesting would generate less pollution than its export. Three constructs were used in the study, which aimed to determine whether the respondents show food neophobia, whether they are willing to seek diversity in food, and what their attitudes towards game meat are. All the scales used were previously validated. Four-hundred and fifty-three questionnaires were collected using the PAPI method. It was found that the respondents showed ambivalent attitudes towards game meat to the greatest extent (76.6%), 16.34% had positive attitudes, and 7.06% had negative attitudes. It seems essential that most of the respondents were highly inclined to look for variety in food (55.85%). Regarding food neophobia, there were 51.43% of people with medium neophobia, while also many people with a low level of neophobia-43.05%. Such results allow speculation that the respondents are open to the new food, they are looking for it, and the low level of game meat consumption is primarily due to the lack of knowledge and awareness about the value of this meat.

Attitudes of Young Tri-City Residents toward Game Meat. Development and Validation of a Scale for Identifying Attitudes toward Wild Meat.

Attitudes toward food are one of the most critical factors related to consumer behavior in the food market. Therefore, identifying attitudes toward a specific food product may be essential for identifying factors influencing certain behaviors regarding game. In addition, game meat is a valuable food that can increase the variety of meat and reduce the intensive breeding of slaughter animals. Therefore, a research gap was found regarding the lack of a tool for identifying attitudes toward game that would allow for the acquisition of data valid for studying conditions related to game consumption. The study aims to validate a developed scale for identifying the attitudes of young Tri-City residents toward game. To collect the database, two groups of respondents are involved in the validation procedure. This procedure includes validation of content, response process, and statistical validation. The scale is validated, and four domains are distinguished based on the PCA test. The validated scale consists of 10 statements (initially 11). The estimated Cronbach's alpha (0.6944) indicates good scale internal consistency. The developed scale can be used to identify attitudes of young Tri-City residents toward game and search for links between these attitudes and behaviors related to game consumption.

Consumer Choices and Habits Related to Tea Consumption by Poles.

Tea is one of the most consumed beverages in the world. In the literature, much attention is paid to the influence of tea and its components on human health and consumer purchasing behavior. The aim of the study was to analyze the habits of Polish consumers regarding tea consumption, brewing methods, and their choices related to tea, to describe the characteristics of tea consumers, and present their segmentation based on consumer choices and habits regarding tea consumption. The study was performed using the computer-assisted web interviewing (CAWI) method on a group of 1700 adult consumers of tea. Information about consumer choices and habits related to tea consumption was collected, including brewing method, place of tea consumption, and factors determining tea choices. Using cluster analysis, six groups of tea consumers were identified. These are "Occasional tea gourmets", "Yerba mate drinkers", "Tea gourmets", "Occasional consumers", "Undemanding tea consumers," and "Occasional strong tea consumers". In summary, it can be said that Poles are not tea gourmets; they prefer black tea, in bags, brewed in cups or glasses for up to 3 min, and usually drink teas without any additives, at home, several times a week, during breakfast and between meals. The most popular brand among the respondents was Lipton. Consumers have little knowledge of the health benefits of tea.

Financing Costs and Health Effects of Air Pollution in the Tri-City Agglomeration.

This paper examines the relationship between the presence of air pollution and incidence of selected respiratory diseases in the urban population of the Tri-City agglomeration. The study takes into consideration the specific character of the region, relating to coastal, and port-based shipping. Three research hypotheses formulated the study. General regression models were used to identify the health effects of air pollution and developed health costs were calculated in relation to the treatment of diseases. The findings have shown that air pollution and climatic conditions in the Tri-City aggravate the symptoms of bronchial asthma, while also increasing the number of cases of exacerbated chronic obstructive pulmonary disease and pneumonia. The evidence demonstrates the negative impact of shipping on the health condition of the inhabitants. The calculations have shown the extent of financial losses incurred in connection with the treatment of diseases found to have been caused by air pollution. The estimated health costs turned out to be significant for each of the examined diseases. The financial inefficiency of the Polish health care system has also been demonstrated. All the models have been identified for monthly data for the first time.

Consumers' Perception and Preference for the Consumption of Wild Game Meat among Adults in Poland.

Wild game meat can be a healthier, safer, and more environmentally friendly alternative to meat from farm animals. The aims of this study were to know the preferences and opinions of Polish consumers regarding game meat and its use in their diet, and to identify consumer segments based on differences in individual game meat choices, concerns, and eating habits related to game meat. The survey was conducted using the platform for online surveys among 1261 adult Poles. Six clusters characterizing the behavior of game consumers were identified (casual consumers, occasional game gourmets, indifferent consumers, occasional consumers, accidental consumers, wild game lovers) and four clusters among those who do not eat game (uninterested, restricted, dislikers, fearful). It has been found that wild game is more often eaten by hunters and their family or friends. The most common reasons for not consuming game are high prices, low availability, no family tradition, and unacceptable taste. Many positive respondents eat game because of its nutritional value but are concerned about the potential health risks and lack of cooking skills. The results of this study indicate the need for information programs for consumers about this meat. They will provide guidance to meat companies about consumer preferences for game and allow them to develop appropriate marketing strategies.

Novel Mouse Models of Bladder Cancer Identify a Prognostic Signature Associated with Risk of Disease Progression.

To study the progression of bladder cancer from non-muscle-invasive to muscle-invasive disease, we have developed a novel toolkit that uses complementary approaches to achieve gene recombination in specific cell populations in the bladder urothelium in vivo, thereby allowing us to generate a new series of genetically engineered mouse models (GEMM) of bladder cancer. One method is based on the delivery of adenoviruses that express Cre recombinase in selected cell types in the urothelium, and a second uses transgenic drivers in which activation of inducible Cre alleles can be limited to the bladder urothelium by intravesicular delivery of tamoxifen. Using both approaches, targeted deletion of the Pten and p53 tumor suppressor genes specifically in basal urothelial cells gave rise to muscle-invasive bladder tumors. Furthermore, preinvasive lesions arising in basal cells displayed upregulation of molecular pathways related to bladder tumorigenesis, including proinflammatory pathways. Cross-species analyses comparing a mouse gene signature of early bladder cancer with a human signature of bladder cancer progression identified a conserved 28-gene signature of early bladder cancer that is associated with poor prognosis for human bladder cancer and that outperforms comparable gene signatures. These findings demonstrate the relevance of these GEMMs for studying the biology of human bladder cancer and introduce a prognostic gene signature that may help to stratify patients at risk for progression to potentially lethal muscle-invasive disease. SIGNIFICANCE: Analyses of bladder cancer progression in a new series of genetically engineered mouse models has identified a gene signature of poor prognosis in human bladder cancer.

Use of Food Services by Consumers in the SARS-CoV-2 Pandemic. How the Eating Habits of Consumers Changed in View of the New Disease Risk Factors?

The SARS-CoV-2 pandemic in 2020-2021 changed the eating habits of people around the world. The aim of this study is to understand the effects of COVID-19 on changing consumers' eating habits, including their concerns about food service nutrition in case of new disease risk factors. The survey conducted using the computer-assisted web-based interviewing method on a group of 1021 adult respondents in Poland. We collected information about consumer choices and habits related to use of food services during the pandemic. This research found that COVID-19 had an impact on consumers' use of food services, both on-site and take-away. Using cluster analysis, we identified five main groups of food service consumers. It was found that almost half of the respondent group did not change their diet during the pandemic, 20% of respondents changed their diet to a positive one, and 20% to a diet that was negative. For respondents the most important forms of protection against COVID-19 in catering establishments were hand disinfection (70.3%), table disinfection (70.4%), wearing of masks and visors by staff (68.2%), and the possibility of cashless payments (64.6%). Based on cluster analysis (eight consumer clusters), we stated that majority of respondents did not see any threats to using catering service during the pandemic. Only a small group (8.1%) of respondents were afraid of the possibility of getting sick with COVID-19. This study presented the effects of COVID-19 on consumer eating behavior in catering and their concerns with food services uses. Discovering consumer concerns can reduce risk, increase food safety and improve eating habits.

Air Pollutants' Concentrations Are Associated with Increased Number of RSV Hospitalizations in Polish Children.

Respiratory syncytial virus (RSV) contributes significantly to pediatric hospitalizations. An association between air pollution and an increased number of RSV cases has been suggested. We sought to evaluate the short-term impact of air pollutants on RSV hospitalizations in Polish children in the period 2010-2019. Daily concentrations of PM10 and PM2.5 (particulate matter with an aerodynamic diameter less than or equal to 10 µm and 2.5 µm, respectively) and nitrogen dioxide (NO2) were analyzed in general regression models (GRM) to establish their influence and full interaction scheme. Significant seasonal and annual periodicity among 53,221 hospitalizations was observed; finally, data from the 2012-2019 RSV high-risk seasons created models for seven agglomerations. The addition of PM2.5, PM10, and NO2 to the basic model for RSV seasonality explained 23% (4.9-31%, univariate model) to 31.4% (8.4-31%, multivariate model) of the variance in RSV hospitalizations. A 10 µg/m3 increase in PM2.5, PM10, and NO2 concentrations was associated with 0.134 (0.087-0.16), 0.097 (0.031-0.087), and 0.212 (0.04-0.29) average increases in hospitalizations, respectively. In the multivariate models, PM2.5, PM10, and NO2 alone, as well as PM2.5-NO2, PM2.5-PM10, and PM10-NO2 interactions, were associated with hospitalizations in some of the locations, while the metaregression showed statistically significant interactions between each of the pollutants, and between the pollutants and the year of the study. The inclusion of PM2.5, PM10, and NO2 in GRM explains a significant number of RSV hospitalizations. The pollutants act alone and interact together in a varied manner. Reducing air contamination might decrease the costs of hospital healthcare.

Consumer Choices and Habits Related to Coffee Consumption by Poles.

Coffee is one of the most popular drinks consumed in the world, also in Poland. In the literature, much attention is paid to the influence of coffee on human health, especially daily intake of caffeine, and also purchasing consumer behavior. There is a lack of research devoted to consumer choices and habits in relation to coffee consumption and brewing method. Therefore, the aim of this study is to describe the characteristics of coffee consumers and present their segmentation based on consumer choices and habits towards coffee consumption. The study was performed using the computer-assisted web interviewing (CAWI) method on a group of 1500 adults respondents in Poland reporting the consumption of coffee. We collected information about consumer choices and habits related to coffee consumption, including brewing method, place of consuming coffee, and factors determining coffee choices. Using cluster analysis, we identified three main groups of coffee consumers. There are "Neutral coffee drinkers", "Ad hoc coffee drinkers", and "Non-specific coffee drinkers". The respondents in the study are not coffee gourmets; they like and consume coffee, but these are often changing choices. To conclude, it can be stated that the Polish coffee consumer prefers conventional methods of brewing coffee (like a "traditionalist") but is open to novelties and new sensory experiences. Based on study results it is possible to know the coffee drinking habits in Poland.

Sulfatide decreases the resistance to stress-induced apoptosis and increases P-selectin-mediated adhesion: a two-edged sword in breast cancer progression.

BACKGROUND: We have previously shown that galactosylceramide (GalCer) affects the tumourigenic and metastatic properties of breast cancer cells by acting as an anti-apoptotic molecule. Since GalCer is a precursor molecule in the synthesis of sulfatides, the present study was aimed to define the role of sulfatides in apoptosis and breast cancer progression. METHODS: Expression of GAL3ST1 in breast cancer cell lines and breast cancer tissue specimens was analysed using real-time PCR, western blotting and immunohistochemistry analysis. The amount of sulfatide, GalCer and ceramide was analysed by thin-layer chromatography binding assay and by the modified hydrophilic interaction liquid chromatography coupled with electrospray mass spectrometry methodology. The tumourigenicity of cancer cells was analysed by an in-vivo tumour growth assay. Apoptotic cells were detected based on caspase-3 activation and the TUNEL assay. The interaction of breast cancer cells with P-selectin or E-selectin was analysed using the flow adhesion assay. The ability of sulfatide-expressing cells to activate and aggregate platelets was studied using the flow-cytometry-based aggregation assay. RESULTS: Using two models of breast cancer, T47D cells with blocked synthesis of sulfatide and MDA-MB-231 cells with neosynthesis of this glycosphingolipid, we showed that high sulfatide levels resulted in increased sensitivity of cancer cells to apoptosis induced by hypoxia and doxorubicin in vitro, and decreased their tumourigenicity after transplantation into athymic nu/nu mice. Accordingly, a clinical study on GAL3ST1 expression in invasive ductal carcinoma revealed that its elevated level is associated with better prognosis. Using MDA-MB-231 cells with neosynthesis of sulfatide we also showed that sulfatide is responsible for adhesion of breast cancer cells to P-selectin-expressing cells, including platelets. Sulfatide also acted as an activating molecule, increasing the expression of P-selectin. CONCLUSIONS: This study demonstrates that increased synthesis of sulfatide sensitises cancer cells to microenvironmental stress factors such as hypoxia and anticancer drugs such as doxorubicin. However, sulfatide is probably not directly involved in apoptotic cascades, because its increased synthesis by GAL3ST1 decreased the amounts of its precursor, GalCer, a known anti-apoptotic molecule. On the other hand, our data support the view that sulfatides are malignancy-related adhesive molecules involved in activating and binding P-selectin-expressing platelets to breast cancer cells.

Tumor Evolution and Drug Response in Patient-Derived Organoid Models of Bladder Cancer.

Bladder cancer is the fifth most prevalent cancer in the U.S., yet is understudied, and few laboratory models exist that reflect the biology of the human disease. Here, we describe a biobank of patient-derived organoid lines that recapitulates the histopathological and molecular diversity of human bladder cancer. Organoid lines can be established efficiently from patient biopsies acquired before and after disease recurrence and are interconvertible with orthotopic xenografts. Notably, organoid lines often retain parental tumor heterogeneity and exhibit a spectrum of genomic changes that are consistent with tumor evolution in culture. Analyses of drug response using bladder tumor organoids show partial correlations with mutational profiles, as well as changes associated with treatment resistance, and specific responses can be validated using xenografts in vivo. Our studies indicate that patient-derived bladder tumor organoids represent a faithful model system for studying tumor evolution and treatment response in the context of precision cancer medicine.

ARF Confers a Context-Dependent Response to Chemotherapy in Muscle-Invasive Bladder Cancer.

Muscle-invasive bladder cancer (MIBC) generally responds poorly to treatment and tends to exhibit significant mortality. Here we show that expression of the tumor suppressor p14ARF (ARF) is upregulated in aggressive subtypes of MIBC. Accumulation of ARF in the nucleolus is associated with poor outcome and attenuated response to chemotherapy. In both genetically engineered mouse models and murine xenograft models of human MIBC, we demonstrate that tumors expressing ARF failed to respond to treatment with the platinum-based chemotherapy agent cisplatin. Resistance was mediated in part by the integrin-binding protein ITGB3BP (CENPR) and reflected ARF-dependent impairment of protein translation, which was exaggerated by drug treatment. Overall, our results highlight a context-dependent role for ARF in modulating the drug response of bladder cancer. Cancer Res; 77(4); 1035-46. ©2017 AACR.

Expression of metallothionein in the liver and kidneys of the red deer (Cervus elaphus L.) from an industrial metal smelting area of Poland.

The metallothionein 1 (MT1) coding sequence of red deer was identified and compared to orthologous sequences from other mammals. Over 90% identity was observed between red deer MT1 amino acid sequence and MT1 sequences of other ruminants. Liver and kidney samples of red deer were collected from the industrial zinc smelting site of Miasteczko Slaskie and from the Masuria Lake District serving as a pollution-free control site. The concentrations of cadmium (Cd), lead (Pb), copper (Cu) and zinc (Zn) were analyzed by the atomic absorption spectrometry technique (AAS). The levels of Cd in the liver of red deer from the metal smelting region was about 8 times higher than for the reference control site. Next, the expression of MT1 mRNA in the liver of red deer was quantified by the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and the expression of MT1/2 protein in the liver and kidneys was analyzed by immunohistochemistry. Positive correlations were found between expression levels for MT1 mRNA and the concentrations of Cu and Zn in liver of red deer, and with the age of animals. Immunohistochemical staining demonstrated the nuclear and cytoplasmatic expression in both liver and kidney tissues, but with no obvious relationship shown for the expression of MT1/2 protein and tissue metal levels. Our results showed that the analysis of MT expression levels in the red deer could not be used independently as a biomarker for identifying exposure to Cd, but could be co-analyzed with tissue metal levels to give better prognosis for environmental exposure to metals.

Elevated PIM2 gene expression is associated with poor survival of patients with acute myeloid leukemia.

The PIM2 gene encodes the serine/threonine kinase involved in cell survival and apoptosis. The aim of the study was to evaluate the expression of the PIM2 gene in acute myeloid leukemia (AML) and to examine its role in apoptosis of the blastic cells. We analyzed the PIM2 expression in 148 patients: 91 with AML, 57 with acute lymphoblastic leukemia and 24 healthy controls by Real-Time PCR and Western blot. Inhibition of the PIM2 gene in human leukemic HL60 cell line was performed with RNAi and apoptosis rate was analyzed. Our results indicate that overexpression of PIM2 in AML is associated with low complete remission rate, high-risk cytogenetics, shorter leukemia-free survival, and event-free survival. Cytometric analysis of HL60/PAC-GFP and HL60/PAC-GFP-shPIM2 cells revealed an increase in the number of apoptotic cells after inhibition of PIM2 gene. In summary, the elevated expression of PIM2 in blastic cells is associated with poor prognosis of AML patients and their resistance to induction therapy.

Metallothionein-3 Increases Triple-Negative Breast Cancer Cell Invasiveness via Induction of Metalloproteinase Expression.

It has been recently found that metallothionein-3 (MT3) enhances the invasiveness and tumorigenesis of prostate cancer cells. This finding is in contrast to those of earlier studies, which indicated that overexpression of MT3 in breast cancer and prostate cancer cell lines inhibits their growth in vitro. Therefore, to clarify the role of MT3 in breast cancer progression, we analyzed the effect of MT3-overexpression on proliferation, invasiveness, migration, and tumorigenesis of breast cancer MDA-MB-231/BO2 cells. It was found that MDA-MB-231/BO2 cells overexpressing MT3 were characterized by increased invasiveness in vitro, compared to the control cells. Interestingly, this increased invasiveness correlated with a highly increased concentration of MMP3 in the culture supernatants (p<0.0001). Our data suggest that MT3 may regulate breast cancer cell invasiveness by modulating the expression of MMP3. These experimental results, obtained using triple-negative MDA-MB-231/BO2 cells, were further supported by clinical data. It was found that, in triple-negative breast cancer (TNBC), nuclear MT3 immunoreactivity in cancer cells tended to be associated with patients' shorter disease-specific survival, suggesting that nuclear MT3 expression may be a potential marker of poor prognosis of triple-negative TNBC cases.

Expression of metallothioneins I and II in kidney of doxorubicin-treated rats.

Metallothioneins I/II (MT) are commonly expressed in mammalian tissues and are highly inducible in the response to stress conditions. Doxorubicin (DOX) intoxication promotes oxidative stress and subsequent apoptosis leading to kidney damage. The present study investigates a correlation between endogenous MT expression and DOX-induced apoptosis in renal tubular cells. Experiments were conducted on Buffalo rats receiving DOX (8 mg/kg b.w. for 3 weeks) versus control rats injected with saline. The histopathological alterations and apoptosis (TUNEL) were evaluated in tissue sections. MT expression and tissue localization was examined using immunohistochemical method (IHC). Western blot (WB) was used to evaluate pro-caspase-3, active caspase-3 and MT expression level in tissue homogenates. Examination of renal tissue revealed severe nephrotoxicity in DOX-treated animals. Apoptosis was observed in distal convoluted tubular cells, whereas MT was detected in proximal tubular cells. A significant increase in pro-caspase-3, active caspase-3 and MT expression levels (WB) were seen in DOX group. Positive correlations between histopathological lesions, apoptosis and MT expression were observed. The results obtained in this study could suggest the protective and antiapoptotic effect of MT expression in renal proximal tubular cells under DOX intoxication.

Modelling bladder cancer in mice: opportunities and challenges.

The prognosis and treatment of bladder cancer have improved little in the past 20 years. Bladder cancer remains a debilitating and often fatal disease, and is among the most costly cancers to treat. The generation of informative mouse models has the potential to improve our understanding of bladder cancer progression, as well as to affect its diagnosis and treatment. However, relatively few mouse models of bladder cancer have been described, and in particular, few that develop invasive cancer phenotypes. This Review focuses on opportunities for improving the landscape of mouse models of bladder cancer.

Nogo-B receptor expression correlates negatively with malignancy grade and ki-67 antigen expression in invasive ductal breast carcinoma.

BACKGROUND: Nogo-B receptor (NgBR) has been shown to be involved in endothelial cell chemotaxis and morphogenesis. However, few studies analyzing its expression in cancer cells have been performed. MATERIALS AND METHODS: We examined NgBR expression in 233 patients with invasive ductal breast carcinoma (IDC) and corresponding non-malignant breast tissues (NMBT) on mRNA (real-time polymerase chain reaction) and protein levels (immunohistochemistry; IHC and western-blot analysis). NgBR expression was found also analyzed in breast cancer cell lines of varying invasiveness. RESULTS: NgBR expression was increased in IDC compared to NMBT on the mRNA (p=0.0007) and protein level (p=0.018). NgBR expression decreased significantly with IDC malignancy grade and correlated negatively with the Ki-67 antigen expression (r=-0.18; p=0.0005). High NgBR mRNA expression was associated with estrogen receptor negativity (p=0.0023) and the triple-negative phenotype of the tumors (p=0.0129). CONCLUSION: NgBR may be involved in IDC development, however, its role in its progression requires further research.