Lyophilized cell-free supernatants of the oral probiotics Streptococcus salivarius M18 and Streptococcus salivarius K12 show promises for milk safety.

The functional food ingredients market has been growing due to the preferences for healthier, nutritional, environment-friendly, and convenience foods. Here, we evaluated the antimicrobial potential of the lyophilized cell-free supernatants of the two most promising oral probiotic strains Streptococcus salivarius M18 and S. salivarius K12 on Pseudomonas aeruginosa to be applied for safety purposes in the milk. We showed that the lyophilized culture supernatant of the strain M18 inhibited the pathogen growth in milk by about 75%, 70%, and 60% when incubated at 37°C, room temperature, and +4°C, respectively. The inhibition levels were about 50%, 30%, and 45% for the lyophilized K12 cell-free supernatant. Besides, the lyophilized culture supernatants of the oral probiotics, especially of S. salivarius M18, exhibited anti-cancer activities on colon cancer cells in vitro. Thus, the results of this manuscript suggest that the cell-free supernatants of the M18 and K12 strains are potential candidates, which merit more investigation for their applications, as biopreservatives in foods and beverages and as anti-cancer biotics for human health.

Non-growth inhibitory doses of dimethyl sulfoxide alter gene expression and epigenetic pattern of bacteria.

Although dimethyl sulfoxide (DMSO) is a widely used solvent in scientific research, drug screening settings, and biomedical applications, its solvent (vehicle) effects on biological processes are overlooked. Using Escherichia coli as a model, we aimed to investigate and evaluate the effects of low-dose DMSO-driven changes in bacterial cells in a comprehensive and multifaceted manner by combining Fourier transform infrared spectroscopy analyses, analytical cell-biology approaches, and high-throughput sequencing. Here, we show that the non-toxic (1.0 and 2.5%, v/v) DMSO doses reduce the cellular levels of reactive oxygen species, change the cellular nucleic acid content and DNA topology, affect the global 5-methylcytosine pattern of the genome, and modulate gene transcription. These results indicate that even at non-toxic concentrations, DMSO is not inert: it can alter validity by changing or masking the assessed activity of the analyte. Besides, this manuscript does not only highlight that the low, non-toxic solvent doses of DMSO impinge on biological processes, including genome structure and function, but also, the high-throughput sequence data obtained during the study offer a platform for future research to elucidate the mechanism of epigenetically regulated genes in bacteria. KEY POINTS: • A clear-cut differentiation between the low-dose DMSO-treated and -untreated bacteria by PCA and LDA. • Drastic alterations in the DNA topology and nucleic acids of DMSO-treated bacteria. • Changes in transcriptome and epigenetic signatures with the low-dose DMSO.

Molecular Detection of Canine Leishmaniasis in Northern Anatolia, Turkiye.

Background: Canin leishmaniasis (CanL), mostly caused by Leishmania infantum, is one of the most important vector-borne diseases in dogs in the Mediterranean region. In this study, we aimed to determine the disease profile in this region by firstly making microscopic and then molecular analyzes in the samples taken from the dogs. Methods: Overall, 112 whole blood samples taken from dogs for clinical applications by a veterinarian in Cankiri between December 2021 and November 2022 were used. After blood collection, both thin and thick drop blood smear preparations were prepared and evaluated for Giemsa staining. L. infantum was investigated by Real time-PCR (RT-PCR) method from all blood samples. Sequence analysis and phylogenetic tree study were performed on positive samples. Results: Both microscopic and RT-PCR analyzes were performed. In both studies, 3 of the 112 samples were positive. Because of the sequence analysis, they were L. infantum. Sequence analysis was performed from the samples found 3 positive. The phylogenetic tree was drawn by making NCBI (National Center for Biotechnology) data entries of the positive samples (Accession numbers: OQ184728, OQ184729, OQ184730). Conclusion: Dogs are important, as they are reservoir of this disease. In this study, 3 (2.7%) positive Leishmaniasis was detected in dogs in Cankiri. Ultimately, this should prompt discussion about new strategies going forward to combat infection caused by Leishmania.

Tapioca starch and skim milk support probiotic efficacy of Lactiplantibacillus plantarum post-fermentation medium against pathogens and cancer cells.

The production of functional foods containing prebiotic ingredients is an area of particular interest and a very promising market with the potential to dominate the food industry. This study aims to explore the potential of starch-based prebiotic tapioca and skim milk, as low-cost and easily accessible food sources and as natural and "clean label" food ingredients on the probiotic activities of Lactiplantibacillus plantarum (formerly Lactobacillus plantarum). The results show that concomitant use of the modified tapioca starch and skim milk promotes the antibacterial and anti-cancer properties of L. plantarum post-fermentation media pointing out how the functionality of probiotic products can be regulated by growth supplements.

Exploiting the Acidic Extracellular pH: Evaluation of Streptococcus salivarius M18 Postbiotics to Target Cancer Cells.

Previously, we showed that the growth, antibiotic resistance, and biofilm formation properties of the pathogens Pseudomonas aeruginosa and Klebsiella pneumonia were tremendously inhibited by the cell-free supernatant of the oral probiotic Streptococcus salivarius M18. These anti-pathogenic activities of the supernatant were more efficient under acidic conditions. The present approach takes advantage of the acidic nature of the tumor microenvironment to evaluate the effect of the S. salivarius M18 postbiotics on colon cancer cells. In both two-dimensional (2D) and three-dimensional (3D) cell culture models, S. salivarius M18 cell-free supernatant showed anti-cancer actions in the pH conditions mimicking the acidity of the tumor. The inhibitory effect was more prominent when the colon cancer cells have been treated with the cell-free supernatant obtained from the inulin incubated S. salivarius M18. The results of this study point out the potential of the S. salivarius M18 functional probiotic products to be used for targeting low pH environments including the unique acidic microenvironment of tumors.

Cell-free supernatant of Streptococcus salivarius M18 impairs the pathogenic properties of Pseudomonas aeruginosa and Klebsiella pneumonia.

M18 strain of Streptococcus salivarius is a bacterial replacement probiotic that has been suggested for use in the oral cavity. Here, we have shown that S. salivarius M18 cell-free supernatant reduced the growth of the two most common human pathogens Pseudomonas aeruginosa and Klebsiella pneumonia and sensitized the pathogenic bacteria to antibiotic. Besides, the supernatant inhibited biofilm formation of P. aeruginosa drastically. For pinpointing the biomolecular changes that occurred in P. aeruginosa incubated with the probiotic supernatant, attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy was used. Unsupervised learning algorithms, principal component analysis (PCA) and hierarchical cluster analysis (HCA), and intensity analyses of individual spectral bands exhibited comprehensive alterations in the polysaccharide and lipid contents and compositions of P. aeruginosa cultivated with S. salivarius M18 cell-free supernatant. These results indicate that S. salivarius M18 has the potential for the prevention or alleviation of different pathogen-induced infections along with the infections of oral pathogens.

Enhanced expression of HNF4α during intestinal epithelial differentiation is involved in the activation of ER stress.

Intestinal epithelial cells are derived from stem cells at the crypts that undergo differentiation into transit-amplifying cells, which in turn form terminally differentiated enterocytes as these cells reach the villus. Extensive alterations in both transcriptional and translational programs occur during differentiation, which can induce the activation of cellular stress responses such as ER stress-related unfolded protein response (UPR) and autophagy, particularly in the cells that are already committed to becoming absorptive cells. Using an epithelial cell model of enterocyte differentiation, we report a mechanistic study connecting enterocyte differentiation to UPR and autophagy. We report that differentiated colon epithelial cells showed increased cytosolic Ca2+ levels and activation of all three pathways of UPR: inositol-requiring enzyme 1 (IRE1), protein kinase RNA-like ER kinase, and activating transcription factor 6 (ATF6) compared to the undifferentiated cells. Enhanced UPR in the differentiated cells was accompanied by the induction of autophagy as evidenced by increased ratio of light chain 3 II/I, upregulation of Beclin-1, and downregulation of p62. We show for the first time that mechanistically, the upregulation of hepatocyte nuclear factor 4α (HNF4α) during differentiation led to increased promoter binding and transcriptional upregulation of two major proteins of UPR: X-box binding protein-1 and ATF6, implicating HNF4α as a key regulator of UPR response during differentiation. Integrating wet-lab with in silico analyses, the present study links differentiation to cellular stress responses, and highlights the importance of transcription factor signaling and cross-talk between the cellular events in the regulation of intestinal cell differentiation.

Butyrate mediated regulation of RNA binding proteins in the post-transcriptional regulation of inflammatory gene expression.

Short chain fatty acids (SCFAs) are produced by commensal bacteria in the gut and are known to reduce inflammation through transcriptional inhibition of cytokines and inflammatory proteins such as cyclooxygenase-2 (COX-2). Butyrate is a SCFA that was reported to alter the mRNA stability of inflammatory genes by increasiing the expression of the RNA binding protein (RBP) Tristetraprolin (TTP). We have hypothesized that butyrate may regulate gene expression post-transcriptionally through global effects on the expression or cytoplasmic translocation of RBPs. Using bioinformatics analyses of publicly available microarray data as well as colon cancer cell lines treated with sodium butyrate, we have observed that butyrate treatment led to a general reduction in expression of several (but not all) RBPs and inhibition in the cytosolic translocation of HuR, a well-known stabilizing RBP. This was reflected in reduced NanoLuc reporter activity of several different AU-rich element (ARE) sequences in the presence of butyrate; this suppression was retained even when HuR was overexpressed. Mechanistically, we have shown that reduced activity of HuR was related to decreased phosphorylation of p38 and MK2 and enhanced phosphorylation of Chk2. As a proof of concept, we show butyrate-mediated inhibition in binding of HuR to the 3'UTR of COX-2 mRNA resulting in reduced mRNA and protein levels of the inflammatory gene. Overall, our data suggest that butyrate can reduce the expression of inflammatory genes not only by transcriptional regulation, but also by post-transcriptional regulation via inhibition of mRNA stabilizing proteins.

Methylation, sugar puckering and Z-form status of DNA from a heavy metal-acclimated freshwater Gordonia sp.

Heavy metal acclimation of bacteria is of particular interest in many aspects. It could add to our understanding of adaptation strategies applied by bacteria, as well as help us in devising ways to use such adaptive bacteria for bioremediation. In this study, we have explored the changes in the DNA of an aquatic Gordonia sp. acclimated to silver, cadmium, and lead. We have measured the changes in the DNA extracted from the acclimated bacteria by using ATR-FTIR coupled with unsupervised and supervised pattern recognition algorithms. Although whole-cell FTIR studies do reveal nucleic acid changes, the special care should be taken when considering marker nucleic acid bands in such spectra, as various other cell or tissue constituents also yield IR bands in the same region. An FTIR study on isolated DNA can be used to avoid this problem. The IR spectral profiles of the DNA molecules revealed significant changes in the backbone and sugar conformations of upon acclimation. We then further analyzed the DNA's global cytosine-methylation patterns of the heavy metal-acclimated bacteria. We aimed to find out whether epigenetic mechanisms operate in bacteria for survival and growth in inhibitory heavy metal concentrations or not. We found hypermethylation in Cd acclimation but hypomethylation for both Pb and Ag in Gordonia sp. Our results imply that changes in the conformational and methylation states of DNA seem to let bacteria to thrive in otherwise inhibitory conditions and mark the involvement of epigenetic modulation in acclimation processes.

Evaluation of colloidal platinum on cytotoxicity, oxidative stress and barrier permeability across the gut epithelium.

Colloidal platinum (Pt) is widely consumed due to its health promoting benefits. However, the exact biological effects of these nanoparticles have not been studied in detail, particularly in the gut. In the present study we observed that colloidal Pt was not cytotoxic towards three different epithelial colon cancer cell lines. Co-treatment of the colon cancer cell line Caco-2 with the oxidative stress inducing agent hydrogen peroxide (H2O2) and colloidal Pt resulted in a significant decrease in H2O2 induced oxidative stress. Colloidal Pt by itself did not induce any oxidative stress. Additionally, both overnight pretreatment of Caco-2 cells with colloidal Pt followed by 1 h treatment with H2O2, or co-treatment of cells for 1 h with colloidal Pt and H2O2 resulted in a significant recovery of cell death. Of note, the same protective effects of colloidal Pt were not observed when the oxidative stress was induced in the presence of 2, 2-azobis (2-amidinopropane) dihydrochloride, indicating that the source of free radicals may define the outcome of anti-oxidant activity of colloidal Pt. Colloidal Pt was also able to cross a model intestinal barrier formed in vitro with differentiated Caco-2 cells easily. Overall, our data indicate that colloidal Pt was not toxic towards intestinal epithelial cells, reduced H2O2 induced oxidative stress, protected from oxidative stress related death of intestinal epithelial cells and could pass a model gut barrier easily. Colloidal Pt can therefore be consumed orally for its anti-oxidant and other health promoting benefits.

Biodegradable polymer promotes osteogenic differentiation in immortalized and primary osteoblast-like cells.

Biodegradable polymers have been broadly used as agents that can complex with and deliver osteoinductive agents, but osteoinductivity of the polymers themselves has been rarely studied. Here we report the osteoinductivity of poly(4-hydroxy-L-proline ester) (PHPE), a biodegradable cationic polymer with cell penetrating properties. Under physiological conditions, PHPE degrades into trans-4-hydroxy-L-proline (trans-Hyp), a non-coded amino acid with essential functions in collagen fibril formation and fibril stability. Treatment of SaOS-2 osteoblast-like cells and hFOB 1.19 primary osteoblast cells with PHPE promoted earlier collagen nodule formation and mineralization of the extracellular matrix compared to untreated cells, even when mineralization activators were absent in the growth medium. Our results indicate that PHPE is a potential osteoinductive agent in vitro that can favor bone regeneration. Moreover, this osteoinductive property could be partly attributed to the degradation product trans-Hyp, which could recapitulate some, but not all of the osteogenic activity. The primary findings of this study can be summarized as follows: treatment of cells with PHPE led to (1) the induction of COL1A1 expression, collagen synthesis and secretion in osteoblast-like cells, (2) mineralization of the ECM in both SaOS-2 and hFOB 1.19 primary osteoblasts, and (3) induction of BMP2 gene and protein expression in osteoblast-like cells, which can promote mineralization of the ECM and regeneration of the bone tissue. Our results suggest that PHPE is a non-cytotoxic polymer and can be potentially used to overcome collagenopathies such as osteogenesis imperfecta.

Determination of Autophagy in the Caco-2 Spontaneously Differentiating Model of Intestinal Epithelial Cells.

The Caco-2 colorectal cancer cell line is widely used as a model for intestinal differentiation and barrier function. These cells, upon reaching confluency, spontaneously differentiate into enterocyte-like cells, synthesize intestinal enzymes, and form domes. Caco-2 cells also undergo autophagy in the course of differentiation. The criteria to establish the induction of autophagy in cells are already well established. Here, we describe the protocol for the spontaneous differentiation of Caco-2 cells and the detection of autophagy using Western blot, flow cytometry, and immunofluorescence.

Low dose dimethyl sulfoxide driven gross molecular changes have the potential to interfere with various cellular processes.

Dimethyl sulfoxide (DMSO) is a small molecule with polar, aprotic and amphiphilic properties. It serves as a solvent for many polar and nonpolar molecules and continues to be one of the most used solvents (vehicle) in medical applications and scientific research. To better understand the cellular effects of DMSO within the concentration range commonly used as a vehicle (0.1-1.5%, v/v) for cellular treatments, we applied Attenuated Total Reflectance (ATR) Fourier Transform Infrared (FT-IR) spectroscopy to DMSO treated and untreated epithelial colon cancer cells. Both unsupervised (Principal Component Analysis-PCA) and supervised (Linear Discriminant Analysis-LDA) pattern recognition/modelling algorithms applied to the IR data revealed total segregation and prominent differences between DMSO treated and untreated cells at whole, lipid and nucleic acid regions. Several of these data were supported by other independent techniques. Further IR data analyses of macromolecular profile indicated comprehensive alterations especially in proteins and nucleic acids. Protein secondary structure analysis showed predominance of ß-sheet over α-helix in DMSO treated cells. We also observed for the first time, a reduction in nucleic acid level upon DMSO treatment accompanied by the formation of Z-DNA. Molecular docking and binding free energy studies indicated a stabilization of Z-DNA in the presence of DMSO. This alternate DNA form may be related with the specific actions of DMSO on gene expression, differentiation, and epigenetic alterations. Using analytical tools combined with molecular and cellular biology techniques, our data indicate that even at very low concentrations, DMSO induces a number of changes in all macromolecules, which may affect experimental outcomes where DMSO is used as a solvent.

Emerging cellular functions of the lipid metabolizing enzyme 15-Lipoxygenase-1.

The oxygenation of polyunsaturated fatty acids such as arachidonic and linoleic acid through lipoxygenases (LOXs) and cyclooxygenases (COXs) leads to the production of bioactive lipids that are important both in the induction of acute inflammation and its resolution. Amongst the several isoforms of LOX that are expressed in mammals, 15-LOX-1 was shown to be important both in the context of inflammation, being expressed in cells of the immune system, and in epithelial cells where the enzyme has been shown to crosstalk with a number of important signalling pathways. This review looks into the latest developments in understanding the role of 15-LOX-1 in different disease states with emphasis on the emerging role of the enzyme in the tumour microenvironment as well as a newly re-discovered form of cell death called ferroptosis. We also discuss future perspectives on the feasibility of use of this protein as a target for therapeutic interventions.

Aspects of silver tolerance in bacteria: infrared spectral changes and epigenetic clues.

In this study, the molecular profile changes leading to the adaptation of bacteria to survive and grow at inhibitory silver concentration were explored. The profile obtained through infrared (IR)-based measurements indicated extensive changes in all biomolecular components, which were supported by chemometric techniques. The changes in biomolecular profile were prominent, including nucleic acids. The changes in nucleic acid region (1350-950 cm-1 ) were encountered as a clue for conformational change in DNA. Further analysis of DNA by IR spectroscopy revealed changes in the backbone and sugar conformations. Moreover, Enzyme-Linked Immunosorbent Assay-based measurements of DNA methylation levels were performed to see if epigenetic mechanisms are in operation during bacterial adaptation to this environmental challenge. The results indicated a notable demethylation in Escherichia coli and methylation in Staphylococcus aureus likely to be associated with their elaborate adaptation process to sustain survival and growth.

Opposing roles of the aldo-keto reductases AKR1B1 and AKR1B10 in colorectal cancer.

PURPOSE: Aldo-keto reductases (including AKR1B1 and AKR1B10) constitute a family of oxidoreductases that have been implicated in the pathophysiology of diabetes and cancer, including colorectal cancer (CRC). Available data indicate that, despite their similarities in structure and enzymatic functions, their roles in CRC may be divergent. Here, we aimed to determine the expression and functional implications of AKR1B1 and AKR1B10 in CRC. METHODS: AKR1B1 and AKR1B10 gene expression levels were analyzed using publicly available microarray data and ex vivo CRC-derived cDNA samples. Gene Set Enrichment Analysis (GSEA), The Cancer Genome Atlas (TCGA) RNA-seq data and The Cancer Proteome Atlas (TCPA) proteome data were analyzed to determine the effect of high and low AKR1B1 and AKR1B10 expression levels in CRC patients. Proliferation, cell cycle progression, cellular motility, adhesion and inflammation were determined in CRC-derived cell lines in which these genes were either exogenously overexpressed or silenced. RESULTS: We found that the expression of AKR1B1 was unaltered, whereas that of AKR1B10 was decreased in primary CRCs. GSEA revealed that, while high AKR1B1 expression was associated with increased cell cycle progression, cellular motility and inflammation, high AKR1B10 expression was associated with a weak inflammatory phenotype. Functional studies carried out in CRC-derived cell lines confirmed these data. Microarray data analysis indicated that high expression levels of AKR1B1 and AKR1B10 were significantly associated with shorter and longer disease-free survival rates, respectively. A combined gene expression signature of AKR1B10 (low) and AKR1B1 (high) showed a better prognostic stratification of CRC patients independent of confounding factors. CONCLUSIONS: Despite their similarities, the expression levels and functions of AKR1B1 and AKR1B10 are highly divergent in CRC, and they may have prognostic implications.

15-Lipoxygenase-1 re-expression in colorectal cancer alters endothelial cell features through enhanced expression of TSP-1 and ICAM-1.

15-lipoxygenase-1 (15-LOX-1) oxygenates linoleic acid to 13(S)-hydroxyoctadecadienoic acid (HODE). The enzyme is widely suppressed in different cancers and its re-expression has tumor suppressive effects. 15-LOX-1 has been shown to inhibit neoangiogenesis in colorectal cancer (CRC); in the present study we confirm this phenomenon and describe the mechanistic basis. We show that re-expression of 15-LOX-1 in CRC cell lines resulted in decreased transcriptional activity of HIF1α and reduced the expression and secretion of VEGF in both normoxic and hypoxic conditions. Conditioned medium (CM) was obtained from CRC or prostate cancer cell lines re-expressing 15-LOX-1 (15-LOX-1CM). 15-LOX-1CM treated aortic rings from 6-week old C57BL/6 mice showed significantly less vessel sprouting and more organized structure of vascular network. Human umbilical vein endothelial cells (HUVECs) incubated with 15-LOX-1CM showed reduced motility, enhanced expression of intercellular cell adhesion molecule (ICAM-1) and reduced tube formation but no change in proliferation or cell-cycle distribution. HUVECs incubated with 13(S)-HODE partially phenocopied the effects of 15-LOX-1CM, i.e., showed reduced motility and enhanced expression of ICAM-1, but did not reduce tube formation, implying the importance of additional factors. Therefore, a Proteome Profiler Angiogenesis Array was carried out, which showed that Thrombospondin-1 (TSP-1), a matrix glycoprotein known to strongly inhibit neovascularization, was expressed significantly more in HUVECs incubated with 15-LOX-1CM. TSP-1 blockage in HUVECs reduced the expression of ICAM-1 and enhanced cell motility, thereby providing a mechanism for reduced angiogenesis. The anti-angiogenic effects of 15-LOX-1 through enhanced expressions of ICAM-1 and TSP-1 are novel findings and should be explored further to develop therapeutic options.

Eicosanoid pathway in colorectal cancer: Recent updates.

Enzymatic metabolism of the 20C polyunsaturated fatty acid (PUFA) arachidonic acid (AA) occurs via the cyclooxygenase (COX) and lipoxygenase (LOX) pathways, and leads to the production of various bioactive lipids termed eicosanoids. These eicosanoids have a variety of functions, including stimulation of homeostatic responses in the cardiovascular system, induction and resolution of inflammation, and modulation of immune responses against diseases associated with chronic inflammation, such as cancer. Because chronic inflammation is essential for the development of colorectal cancer (CRC), it is not surprising that many eicosanoids are implicated in CRC. Oftentimes, these autacoids work in an antagonistic and highly temporal manner in inflammation; therefore, inhibition of the pro-inflammatory COX-2 or 5-LOX enzymes may subsequently inhibit the formation of their essential products, or shunt substrates from one pathway to another, leading to undesirable side-effects. A better understanding of these different enzymes and their products is essential not only for understanding the importance of eicosanoids, but also for designing more effective drugs that solely target the inflammatory molecules found in both chronic inflammation and cancer. In this review, we have evaluated the cancer promoting and anti-cancer roles of different eicosanoids in CRC, and highlighted the most recent literature which describes how those molecules affect not only tumor tissue, but also the tumor microenvironment. Additionally, we have attempted to delineate the roles that eicosanoids with opposing functions play in neoplastic transformation in CRC through their effects on proliferation, apoptosis, motility, metastasis, and angiogenesis.

Single nucleotide polymorphisms in the 3'UTR of VPAC-1 cooperate in modulating gene expression and impact differently on the interaction with miR525-5p.

Complex immune and neurodegenerative disorders are the result of multiple interactions between common genetic variations having, individually, a weak effect on the disease susceptibility or resistance. Interestingly, some genes have been found to be associated with more than one disease although not necessarily the same SNPs are involved. In this context, single nucleotide polymorphisms in the 3'UTR region of type 1 receptor (VPAC-1) for vasoactive intestinal peptide (VIP) have been reported to be associated with some immune-mediated as well as with neurodegenerative diseases such as Alzheimer's Disease (AD). Here, we demonstrate that variations at the 3'UTR of the VPAC-1 gene act synergistically to affect the expression of the luciferase as well as of the GFP reporter genes expressed in HEK293T cells. Moreover, the miRNA 525-5p, previously shown by us to target the 3'UTR of VPAC-1, is more efficient in decreasing GFP expression when co-expressed with constructs carrying the allele C at rs896 (p<10(-3)) suggesting that this miRNA regulates VPAC-1 expression at different levels depending on rs896 polymorphism and thus adding complexity to the network of disease susceptibility.

The multifaceted nature of NLRP12.

NLRs are a class of cytoplasmic PRRs with various functions, ranging from pathogen/damage sensing to the modulation of inflammatory signaling and transcriptional control of MHC and related genes. In addition, some NLRs have been implicated in preimplantation and prenatal development. NLRP12 (also known as RNO, PYPAF7, and Monarch-1), a member of the family containing an N-terminal PYD, a NBD, and a C-terminal LRR region, is one of the first described NLR proteins whose role remains controversial. The interest toward NLRP12 has been boosted by its recent involvement in colon cancer, as well as in the protection against some severe infections, such as that induced by Yersinia pestis, the causative agent of plague. As NLRP12 is mainly expressed by the immune cells, and its expression is down-regulated in response to pathogen products and inflammatory cytokines, it has been predicted to play a role as a negative regulator of the inflammatory response. Herein, we present an overview of the NLR family and summarize recent insights on NLRP12 addressing its contribution to inflammatory signaling, host defense, and carcinogenesis.