N-nitroso Compounds Research Articles

In-situ formation and evaluation of N-nitrosamine drug substance related impurities in glycopeptides implying orbitrap mass spectrometry

Nitrosamines, a class of N-nitroso compounds, have raised significant health concerns due to their established carcinogenicity. ICH M7 enlisted N-nitroso compounds in the so called cohorts of concern due to their carcinogenic effects. Glycopeptides (GPs) are complex molecules composed of peptide and glycan moieties. GPs serve as the last resort for the mitigation of bacterial infections, particularly against gram-positive bacteria. GPs are susceptible to nitrosamine drug substance related impurities (NDSRI) contamination due to the presence of secondary amine in their core structure. The incidence of formation of NDSRI impurities in GPs could be either during their semi-synthetic route or storage. However, till date, no studies have been reported on the occurrence of NDSRI in GPs which is a pre-requisite. Hence, the current study investigates the plausible mechanisms and detection methods for the NDSRI in GPs. In-situ studies were performed to evaluate the possible formation of NDSRI in GPs. In the current study, GPs of different generations were screened for their potential of forming NDSRI impurities implying nitrosating agent. LC and LCHRMS/MS studies were performed to identify the in-situ generated impurities. Interestingly, the formation of NDSRIs is evident in all the selected GPs. The molecular mechanisms and pathways for individual GPs and respective NDSRIs were elucidated. Interestingly, isomeric NDSRIs were also identified during in-situ generated samples. Using CPCA the potency scores and acceptable intakes for the NDSRIs were evaluated. Thus, this work aims to enhance the safety and efficacy of GPs ensuring compliance with the regulatory standards by advancing the understanding of NDSRIs in GPs.

Limosilactobacillus fermentum CGMCC 1.7434 and Debaryomyces hansenii GDMCC 2.149 synergize with ultrasound treatment to efficiently degrade nitrite in air-dried ducks

Nitrites in meat products are important food additives with coloring, antibacterial and antioxidant effects, but excessive intake of nitrites can pose health risks, including an increased risk of cancer due to the formation of carcinogenic nitrosamines. In the present study, Limosilactobacillus fermentum CGMCC 1.7434 was screened and the effects of it and Debaryomyces hansenii GDMCC 2.149 and their combination on nitrite degradation were investigated. It was found that the co-culture of L. fermentum CGMCC 1.7434 and D. hansenii GDMCC 2.149 significantly enhanced nitrite degradation (99.58%). The findings on salt and ethanol tolerance suggest suitability for application in meat fermentation processes. Scanning electron microscopy and additional data indicate that D. hansenii GDMCC 2.149 facilitates the growth, acid production, adhesion, secretion of AI-2 signaling molecules, and biofilm formation of L. fermentum CGMCC 1.7434. Metabolomics analysis suggests that these microorganisms reduce nitrite levels by converting NH3 derived from nitrite into L-glutamine, which is further transformed into N-nitroso compounds and their downstream derivatives through the ABC transporter pathway, the TCA cycle, and the amino acid metabolism pathway. Microbial community analyses showed that L. fermentum CGMCC 1.7434 and D. hansenii GDMCC 2.149 were successfully inoculated into air-dried ducks, becoming dominant strains and effectively inhibiting the growth of pathogenic bacteria. Furthermore, during the processing of air-dried duck, the combination of ultrasonic cavitation (250 W, 4 min, 30°C, 40 kHz) with the co-fermentation of L. fermentum CGMCC 1.7434 and D. hansenii GDMCC 2.149 effectively reduced nitrite content (84.55%) and TVB-N levels in the meat, without compromising color or TBARS values. This is crucial for understanding the mechanism of nitrite degradation by LAB in synergy with yeast and for the advancement of low-nitrite air-dried duck products.

NDMA enhances claudin-1 and -6 expression viaCYP2E1/ROS in AGS cells

Carcinogenic N-nitroso compounds, especially N-nitroso dimethylamine, increase the risk of gastric cancer development. Cytochrome P450-2E1 metabolizes this compound, thus generating an oxidant microenvironment. We aimed to evaluate in gastric adenocarcinoma cells if its effect on CYP2E1 and ROS affects signaling pathways associated with gastric cancer oncogenesis. The impact of N- nitroso dimethylamine upon CYP2E1 and ROS activation/secretion was evaluated by the DCFDA assay protocol, TER measurements, Stat3, pSTAT3, ERK1/2, and pERK1/2 expression, claudins-1 and -6 expression, and finally mRNA values of IL-1β IL-6, IL-8 and TNFα. Our results showed that exposure to N- N-nitroso dimethylamine disrupts the regulation of Stat3 and Erk1/2, alters the expression of claudin-1 and claudin-6 tight junction proteins, and increases the secretion of pro-inflammatory cytokines. These alterations induce a continuous local inflammatory process, an event identified as a gastric cancer promoter. In summary, N-nitroso dimethylamine can disrupt cell mechanisms associated with gastric cancer oncogenesis.

Dietary Nitrosamines from Processed Meat Intake as Drivers of the Fecal Excretion of Nitrosocompounds.

Diet is one of the main exogenous sources of potentially carcinogenic nitrosamines (NAs) along with tobacco and cosmetics. Several factors can affect endogenous N-nitroso compounds (NOCs) formation and therefore the potential damage of the intestinal mucosa at initial colorectal cancer stages. To address this issue, 49 volunteers were recruited and classified according to histopathological analyses. Lifestyle and dietary information were registered after colonoscopy. The mutagenicity of fecal supernatants was assayed by a modified Ames test. Fecal heme-derived NOCs and total NOC concentrations were determined by selective denitrosation and chemiluminescence-based detection. Results revealed processed meats as the main source of dietary nitrites and NAs, identifying some of them as predictors of the fecal concentration of heme-derived and total NOCs. Furthermore, increased fecal NOC concentrations were found as the severity of colonic mucosal damage increased from the control to the adenocarcinoma group, these concentrations being strongly correlated with the intake of the NAs N-nitrosodimethylamine, N-nitrosopiperidine, and N-nitrosopyrrolidine. Higher fecal NOC concentrations were also noted in higher fecal mutagenicity samples. These results could contribute to a better understanding of the importance of modulating dietary derived xenobiotics as related with their impact on the intestinal environment and colonic mucosa damage.

Food process contaminants: formation, occurrence, risk assessment and mitigation strategies - a review.

Thermal treatment of food can lead to the formation of potentially harmful chemicals, known as process contaminants. These are adventitious contaminants that are formed in food during processing and preparation. Various food processing techniques, such as heating, drying, grilling, and fermentation, can generate hazardous chemicals such as acrylamide (AA), advanced glycation end products (AGEs), heterocyclic aromatic amines (HAAs), furan, polycyclic aromatic hydrocarbons (PAHs), N-nitroso compounds (NOCs), monochloropropane diols (MCPD) and their esters (MCPDE) which can be detrimental to human health. Despite efforts to prevent the formation of these compounds during processing, eliminating them is often challenging due to their unknown formation mechanisms. It is critical to identify the potential harm to human health in processed food and understand the mechanisms by which harmful compounds form during processing, as prolonged exposure to these toxic compounds can lead to health problems. Various mitigation strategies, such as the use of diverse pre- and post-processing treatments, product reformulation, additives, variable process conditions, and novel integrated processing techniques, have been proposed to control these food hazards. In this review, we summarize the formation and occurrence, the potential for harm to human health produced by process contaminants in food, and potential mitigation strategies to minimize their impact.

Source-specific nitrate intake and all-cause mortality in the Danish Diet, Cancer, and Health Study

IntroductionNitrate and nitrite are naturally occurring in both plant- and animal-sourced foods, are used as additives in the processing of meat, and are found in water. There is growing evidence that they exhibit a spectrum of health effects, depending on the dietary source. The aim of the study was to examine source-dependent associations between dietary intakes of nitrate/nitrite and both all-cause and cause-specific mortality.MethodsIn 52,247 participants of the Danish Diet, Cancer and Health Study, associations between source-dependent nitrate and nitrite intakes––calculated using comprehensive food composition and national drinking water quality monitoring databases––and all-cause, cardiovascular disease (CVD)-related, and cancer-related mortality over 27 years were examined using restricted cubic splines within Cox proportional hazards models adjusting for demographic, lifestyle, and dietary confounders. Analyses were stratified by factors hypothesised to influence the formation of carcinogenic N-nitroso compounds (namely, smoking and dietary intakes of vitamin C, vitamin E, folate, and polyphenols).ResultsPlant-sourced nitrate intake was inversely associated with all-cause mortality [HRQ5vsQ1: 0.83 (0.80, 0.87)] while higher risks of all-cause mortality were seen for higher intakes of naturally occurring animal-sourced nitrate [1.09 (1.04, 1.14)], additive permitted meat-sourced nitrate [1.19 (1.14, 1.25)], and tap water-sourced nitrate [1.19 (1.14, 1.25)]. Similar source-dependent associations were seen for nitrite and for CVD-related and cancer-related mortality except that naturally occurring animal-sourced nitrate and tap water-sourced nitrate were not associated with cancer-related mortality and additive permitted meat-sourced nitrate was not associated with CVD-related mortality. No clear patterns emerged in stratified analyses.ConclusionNitrate/nitrite from plant sources are inversely associated while those from naturally occurring animal-sources, additive-permitted meat sources, and tap water-sources are positively associated with mortality.

The demethylase ALKBH5 mediates ZKSCAN3 expression through the m6A modification to activate VEGFA transcription and thus participates in MNNG-induced gastric cancer progression

N-Nitroso compounds (NOCs) are recognized as important factors that promote gastric cancer development, but the specific effects and potential mechanisms by which NOC exposure promotes gastric cancer are still poorly understood. In this study, we explored the effects and potential molecular mechanisms of NOCs on the promotion of gastric cancer using methylnitronitrosoguanidine (MNNG), a classical direct carcinogen of NOC. The results of in vivo and in vitro experiments showed that chronic and low-concentration MNNG exposure significantly promoted the malignant progression of tumors, including cell migration, cell invasion, vasculogenic mimicry (VM) formation, cell spheroid formation, stem cell-like marker expression, and gastric cancer growth and metastasis. Mechanistically, we revealed that demethylase ALKBH5 regulated the level of the N6‑methyladenosine (m6A) modification in the 3′UTR and CDS region of the ZKSCAN3 mRNA to promote ZKSCAN3 expression, mediated the binding of ZKSCAN3 to the VEGFA promoter region to regulate VEGFA transcription, and participated in MNNG-induced gastric cancer cell migration, invasion, VM formation, cell spheroid formation, stem cell-like marker expression and ultimately gastric cancer progression. In addition, our study revealed that ALKBH5-ZKSCAN3-VEGFA signaling was significantly activated during MNNG-induced gastric carcinogenesis, and further studies in gastric cancer patients showed that ALKBH5, ZKSCAN3, and VEGFA expression were upregulated in cancers compared with paired gastric mucosal tissues, that ALKBH5, ZKSCAN3, and VEGFA could serve as important biomarkers for determining patient prognosis, and that the molecular combination showed greater prognostic value. These findings provide a theoretical basis for developing gastric cancer interventions for NOCs and for determining gastric cancer progression.

Creating a Framework for Treating Autoimmune Gastritis-The Case for Replacing Lost Acid.

Autoimmune gastritis (AIG) is characterized by the destruction of gastric parietal cells, resulting in hypochlorhydria and eventual achlorhydria, as oxyntic glands in the corpus are destroyed and become atrophic. The permanent loss of gastric acid has many impacts-both theoretical and documented. The most concerning of these are hypergastrinemia and increased N-nitroso compounds, both of which increase the risk of gastric cancers. While known deficiencies of B12 and iron are often replaced in AIG, acid is not. Moreover, patients with AIG are often prescribed acid suppression for a stomach that is decidedly no longer acidic, worsening the sequelae of gastric atrophy. Betaine hydrochloride (BHCL) is a short-acting acidifying agent, available over the counter in capsule form. Mealtime acid supplementation has an historic basis and could ameliorate many AIG-related gastrointestinal symptoms. Theoretically, acidification could also reduce the potential for hypergastrinemia and the production of N-nitroso compounds, consequently reducing the risk of gastric cancers. Supplemental vitamin C may also help in preventing gastric N-nitroso formation, regardless of the gastric pH. This narrative review describes the functions of gastric acid in gastrointestinal and immune health, documents the effects of hypochlorhydria in AIG, and proposes potential options for safely re-establishing the acid milieu of the stomach for patients with AIG.

Physicochemical properties and sensory features of ripened, industrially prepared sausages, enriched with olive leaf extract to replace nitrite and nitrate

The use of nitrite and nitrate in meat products has been questioned due to the generation of N-nitroso compounds with carcinogenic effects. Thus, research has been focused on their replacement with several alternatives. This study is focused on evaluating the effects of a commercial olive leaf extract (OLE), marketed as dietary supplement, on protein and lipid oxidation, N-nitrosamine content and sensory properties of industrial sausages at the end of the ripening period. Five different formulations were prepared: F1 (OLE: 1000 mg/kg; NO2–NO3: 0 mg/kg), F2 (OLE: 1000 mg/kg; NO2–NO3: 75-75 mg/kg), F3 (OLE: 0 mg/kg; NO2–NO3: 0 mg/kg), F4 (OLE: 0 mg/kg; NO2–NO3: 75-75 mg/kg), F5 (OLE: 500 mg/kg; NO2–NO3: 35-35 mg/kg). The use of OLE in combination with a reduced dose of nitrite and nitrate (F5) significantly reduced the lipid and protein oxidation (p < 0.05) compared to the formulation with synthetic additives alone (F4). No differences were observed in terms of textural properties with the samples without the extract. The OLE did not affect the microbiological quality and enabled a significant reduction in the N-nitrosamines content. However, the addition of OLE decreased the intensity of red colour and determined the insurgence of a slight bitter taste in the sausages.

N-Nitrosomorpholine-induced oncocytic transformation in rat endocrine organs

BackgroundN-Nitrosomorpholine (NMO) is one of the most common N-nitroso compounds. An oncocytic transformation has been demonstrated in renal tubules of NMO-treated rats. In our study, we aimed to investigate the potential transformation of oncocytic cells in 6 endocrine organs, i.e., thyroid, adrenal and pituitary glands, pancreas, testis, and bone, of NMO-exposed rats.MethodsThirty male rats were born and raised. Fifteen of them were given a single dose of 320 mg NMO per kg body weight, dissolved in drinking water, by a gavage tube. At the end of 52 weeks, the animals in both series were killed. Right after the killing, 6 different endocrine organs (hypophysis, thyroid, pancreas, adrenal gland, bone [femur], and testicles) of each animal were excised.ResultsThere was no evidence of oncocytic cell development in the control group. In contrast, oncocytes were observed in 8 out of 13 NMO-treated rats: 2 in the adrenal sections, 1 in the thyroid sections, 3 in the pituitary sections, and 2 in the pancreas sections. Thesticle and bone sections were completely normal.ConclusionsWe showed that NMO induced an oncocytic change in pancreas, thyroid, pituitary, and adrenal glands. To date, no identified specific environmental risk factors that lead to an oncocytic transformation in endocrine glands have been reported previously. Given the increasing prevalence of endocrine-disrupting chemicals in the environment, personal care products, manufactured goods, and food sources, there is a need to advance our understanding of the pathological mechanisms underlying oncocytosis in endocrine organs.

Nitrate in drinking water and pregnancy outcomes: A narrative review of epidemiological evidence and proposed biological mechanisms

There is emerging evidence of an association between nitrate contamination in drinking water and adverse pregnancy outcomes. Few studies have discussed the evidence in the context of plausible biological mechanisms. We performed a narrative review of the current evidence investigating associations between nitrate in drinking water and the risk of adverse pregnancy outcomes with a focus on congenital anomalies (CA) and preterm birth (PTB). We also reviewed evidence and discuss several biological mechanisms that may explain the observed associations. We reviewed cohort and case-control epidemiological studies assessing associations between maternal nitrate exposure and adverse pregnancy outcomes published until January 2023. Three plausible mechanisms may explain how maternal exposure to nitrate may impact pregnancy outcomes, which include the formation of N-nitroso compounds (NOC), thyroid dysfunction and oxidative stress. For CA, seven studies were included (two cohort and five case-control studies). The strongest associations observed in epidemiological studies were for neural tube defects. Inconsistent positive and negative associations were observed for anomalies of the limb, eye, ear, face and neck. Of the four PTB studies (all cohort studies), three of the largest cohort studies observed associations between PTB with nitrate levels &gt;5 mg/L while one small cohort study did not. Emerging epidemiological evidence has observed a potential increased risk of adverse pregnancy outcomes with elevated maternal nitrate exposure in drinking water. There are also plausible biological mechanisms to support this association. However, the current evidence lacks homogeneity and additional longitudinal evidence with robust exposure asssessments is required. Given the increasing concentrations of nitrate contamination in drinking water in many countries, and the adverse effects observed at concentrations below the current regulatory standard, a further precautionary approach should be adopted until futher evidence emerges.

Ingested nitrate and nitrite and end-stage renal disease in licensed pesticide applicators and spouses in the Agricultural Health Study

BackgroundNitrate and nitrite ingestion has been linked to kidney cancer, possibly via the endogenous formation of carcinogenic N-nitroso compounds. These exposures might also contribute to end-stage renal disease (ESRD).ObjectivesWe investigated associations of drinking water nitrate and dietary nitrate and nitrite intakes (total and by food type) with incident ESRD in the Agricultural Health Study. We also explored modifying effects of vitamin C and heme iron intake, which may affect endogenous nitrosation.MethodsWe performed complete case analyses among private pesticide applicators and their spouses. We obtained water nitrate estimates for participants whose primary drinking water source at enrollment (1993−1997) was public water supplies (PWS) or private wells (N = 59,632). Average nitrate concentrations were computed from historical data for PWS users and predicted from random forest models for private well users. Analysis of dietary nitrate and nitrite was restricted to the 30,177 participants who completed the NCI Dietary History Questionnaire during follow-up (1999−2003). Incident ESRD through 2018 was ascertained through linkage with the U.S. Renal Data System. We estimated adjusted hazard ratios (HRs) and 95%CI for associations of tertiles (T) of exposure with ESRD overall and explored effects in strata of vitamin C and heme iron intake.ResultsWe identified 469 incident ESRD cases (206 for dietary analysis). Water nitrate and total dietary nitrate/nitrite were not associated with ESRD, but increased ESRD was associated with nitrate and nitrite from processed meats. We found apparent associations between nitrite and ESRD only among participants with vitamin C <median (T3 vs. T1 HR: 2.26, 95%CI: 1.05, 4.86) and with heme iron ≥median (T3 vs. T1 HR: 1.73, 95%CI: 0.89, 3.39).SignificanceESRD incidence was associated with dietary nitrate/nitrite from processed meat among all study participants and with total dietary nitrite among participants with lower vitamin C or higher heme iron intake.Impact statementThere are few well-established environmental risk factors for end-stage renal disease (ESRD), a worldwide public health challenge. Ingestion of nitrate and nitrite, which may lead to endogenous formation of carcinogenic N-nitroso compounds, has been linked to some cancers and chronic diseases. We investigated these exposures in relation to ESRD in an agricultural cohort. ESRD incidence was associated with dietary nitrate/nitrite from processed meat and with total dietary nitrite among subgroups with lower vitamin C or higher heme iron intake. This study provides preliminary evidence that points to dietary nitrite and possibly dietary nitrate intake as a potential contributor to ESRD.

Assessment of residual nitrite levels in cooked sausages: Compliance, thermal processing effects, and consumer safety

The increasing global consumption of processed meat, which often contains nitrite as a preservative, raises health concerns due to potential adverse effects from its metabolites, such as nitric oxide and N-nitroso compounds. The study sought to evaluate the food safety of processed meat products within the Serbian market, specifically in the Belgrade region. Nitrite levels were analysed in cooked sausages, both in their raw state and after undergoing the manufacturer's recommended thermal processing prior to consumption, if applicable. Additionally, thus far, there has been a lack of research exploring the potential influence of residual nitrite levels in the meat products prepared as per manufacturer recommendations prior to the consumption of meat products, as well as their contribution to acceptable daily intake (ADI), which provides crucial insights into the overall dietary safety of processed meats. During a three-year period, the study performed analysis on a total of 77 cooked sausages, following the standard ISO methodology. Boiling the cooked sausages led to a major reduction in this meatproduct additive, whereas frying led to a comparatively smaller decrease in nitrite concentration. Additionally, the greatest exposure to nitrite compounds occurs when consuming meat products without prior preparation, i.e., without thermal treatment by consumers before ingestion. In summary, the assessment of the ADI for nitrites revealed a high level of food safety, with all values noticeably below the maximum permitted levels specified by national legislation (150 mg/kg).

Microbiota and detrimental protein derived metabolites in colorectal cancer.

Colorectal cancer (CRC) is the third leading cancer in incidence and the second leading cancer in mortality worldwide. There is growing scientific evidence to support the crucial role of the gut microbiota in the development of CRC. The gut microbiota is the complex community of microorganisms that inhabit the host gut in a symbiotic relationship. Diet plays a crucial role in modulating the risk of CRC, with a high intake of red and processed meat being a risk factor for the development of CRC. The production of metabolites derived from protein fermentation by the gut microbiota is considered a crucial element in the interaction between red and processed meat consumption and the development of CRC. This paper examines several metabolites derived from the bacterial fermentation of proteins associated with an increased risk of CRC. These metabolites include ammonia, polyamines, trimethylamine N-oxide (TMAO), N-nitroso compounds (NOC), hydrogen sulphide (H2S), phenolic compounds (p-cresol) and indole compounds (indolimines). These compounds are depicted and reviewed for their association with CRC risk, possible mechanisms promoting carcinogenesis and their relationship with the gut microbiota. Additionally, this paper analyses the evidence related to the role of red and processed meat intake and CRC risk and the factors and pathways involved in bacterial proteolytic fermentation in the large intestine.

Diethylnitrosamine-Induced Liver Tumorigenesis in Mice Under High-Hat High-Sucrose Diet: Stepwise High-Resolution Ultrasound Imaging and Histopathological Correlations.

The hepatotoxic N-nitroso compound diethylnitrosamine (DEN) administered intraperitoneally (i.p.) induces liver neoplasms in rodents that reproducibly recapitulate some aspects of human hepatocarcinogenesis. In particular, DEN drives the stepwise formation of pre-neoplastic and neoplastic (benign or malignant) hepatocellular lesions reminiscent of the initiation-promotion-progression sequence typical of chemical carcinogenesis. In humans, the development of hepatocellular carcinoma (HCC) is also a multi-step process triggered by continuous hepatocellular injury, chronic inflammation, and compensatory hyperplasia that fuel the emergence of dysplastic liver lesions followed by the formation of early HCC. The DEN-induced liver tumorigenesis model represents a versatile preclinical tool that enables the study of many tumor development modifiers (genetic background, gene knockout or overexpression, diets, pollutants, or drugs) with a thorough follow-up of the multistage process on live animals by means of high-resolution imaging. Here, we provide a comprehensive protocol for the induction of hepatocellular neoplasms in wild-type C57BL/6J male mice following i.p. DEN injection (25mg/kg) at 14days of age and 36weeks feeding of a high-fat high-sucrose (HFHS) diet. We emphasize the use of ultrasound liver imaging to follow tumor development and provide histopathological correlations. We also discuss the extrinsic and intrinsic factors known to modify the course of liver tumorigenesis in this model.

Enriching Cured Meat Products with Bioactive Compounds Recovered from Rosa damascena and Rosmarinus officinalis L. Distillation By-Products: The Pursuit of Natural Antimicrobials to Reduce the Use of Nitrites

Endogenously formed N-nitroso compounds (NOCs) from nitrite use in red meat have been recently linked to the risk of colorectal cancer. Therefore, replacing sodium nitrite (E250) with natural antimicrobials, such as bioactive compounds, is an issue of great industrial and scientific interest. In this research, such compounds were extracted from rose petal (Rosa damascena) and rosemary leaf (Rosmarinus officinalis L.) by-products of the essential oil industry and appropriately introduced in commercial cured meat products, as partial nitrite replacers. Shelf-life tests demonstrated an inhibitory effect of the rose extracts on microbial growth, obtaining similar or reduced growth rates and a prolonged lag phase, when compared to conventionally produced samples (CNT): μmax,ref = 0.128 vs. 0.166 d−1 and λref = 15.9 vs. 12.4 d at 4 °C, respectively. None of the bacon samples suffered from lipid oxidation in the examined period. The RSE samples (containing the rose extract and half of the nitrites) exhibited a good retention of their red color, receiving an acceptable sensory score throughout storage. Since the proposed partial nitrite replacement leads to an important shelf-life extension, namely 78 d (RSE) vs. 60 d (CNT), our results provide preliminary proof of the potential use and valorization of such side streams as effective natural antimicrobial agents for cured meat products, in order to reduce the use of nitrites.

The influence of Helicobacter pylori, proton pump inhibitor, and obesity on the gastric microbiome in relation to gastric cancer development

Helicobacter pylori infection is still the main risk factor for the development of gastric cancer (GC). We explore the scientific evidence for the role of the gastric microbiome beyond Helicobacter pylori (H. pylori) in gastric carcinogenesis. The composition of the gastric microbiome in healthy individuals, in presence and absence of H. pylori infection, in proton pump inhibitor (PPI)-users, obese individuals, and GC patients was investigated. Possible mechanisms for microbial involvement, limitations of available research and options for future studies are provided.A common finding amongst studies was increased levels of Streptococcus, Prevotella, Neisseria, and Actinomyces in healthy individuals or those with H. pylori-negative gastritis. In PPI-users the risk for GC increases with the treatment duration, and the gastric microbiome shifts, with the most consistent increase in the genus Streptococcus. Similarly, in obese individuals, Streptococcus was the most abundant genus, with an increased risk for cardia GC. The genera Streptococcus, Lactobacillus and Prevotella were found to be more prominent in GC patients in multiple studies. Potential mechanisms of non-H. pylori microbiota contributing to GC are linked to lipopolysaccharide production, contribution to inflammatory pathways, and the formation of N-nitroso compounds and reactive oxygen species.In conclusion, the knowledge of the gastric microbiome in GC is mainly descriptive and based on sequencing of gastric mucosal samples. For a better mechanistic understanding of microbes in GC development, longitudinal cohorts including precancerous lesions, different regions in the stomach, and subtypes of GC, and gastric organoid models for diffuse and intestinal type GC should be employed.

Epidemiology of Gastric Cancer: Global Trends, Risk Factors and Premalignant Conditions.

This review article aims to provide a comprehensive overview of recent epidemiology, pathogenesis, risk factors, and premalignant conditions of gastric cancer. Worldwide, gastric cancer is one of the most common and most fatal cancers. The incidence and mortality remain high in regions such as East Asia and Eastern Europe. Although there is a lower incidence in the United States, it remains a deadly disease. Age, gender, and race are non-modifiable demographic risk factors for developing gastric cancer. There have been several dietary and lifestyle risk factors such as salt preserved foods, N-nitroso compounds containing foods, tobacco smoke, alcohol use, and obesity that have been shown to contribute to the development of gastric cancer. Infections have additionally been shown to have a clear role in the pathogenesis of gastric cancer as Helicobacter pylori eradication has shown a significant reduction in the incidence of gastric cancer as well as other pathogens such as Epstein-Barr virus. There are certain premalignant lesions that increase the risk of developing gastric cancer. These include atrophic gastritis, and intestinal metaplasia amongst others.

Characterization and Quantitation of N-Nitroso Duloxetine Impurity in Duloxetine Hydrochloride Drug Substance

The nitrosamine family of chemicals is a large group of carcinogens found in human diets and other environmental media. In animal experiments, it has been shown that N-nitroso compounds can cause cancer, hence the presence of nitrosamine impurities are important to evaluate and need to be quantified. This work shows the content of N-nitroso duloxetine in duloxetine (DXT) hydrochloride, before proceeding to the quantification of impurity, the impurity standard must be well characterized by analytical techniques and it places a vigorous role, i.e. N-nitroso DXT molecular weight conformed by mass spectral data and the atoms connectivity was confirmed by NMR spectral data (1H, 13C, COSY, HSQC and HMBC) and thermal analyses (DSC and TGA) were also performed. In N-nitroso DXT, two sets of signals were typically observed, indicating the presence of asymmetric N-nitroso DXT, concluding with drift functional theory (DFT) for E-, Z-isomers related to the nitroso group in N-nitroso DXT. The medical agency recommends a specified limit of 0.83 ppm for the acceptable intake (AI) and the maximum daily dosage. The determination of N- itroso DXT was performed by liquid chromatography mass spectrometry (LC-MS) followed by electron spray ionization with triple quadrupole.

LDHA and LDHB overexpression promoted the Warburg effect in malignantly transformed GES-1 cells induced by N-nitroso compounds.

N-nitroso compounds (NOCs) exposure is a major risk factor for the development of gastric cancer. However, the carcinogenic mechanisms by which NOCs induce gastric and other cancers, especially the NOCs-induced Warburg effect, have not been comprehensively studied. Lactate dehydrogenase (LDH), which has two subunits (LDHA and LDHB), plays an important role in the Warburg effect of tumor cells. Therefore, we hypothesized that LDHA and LDHB could promote Warburg effect in malignant transformed GES-1cells induced by Nmethyl-N'-nitro-N-nitrosoguanidine (MNNG). GES-1cells were exposed to 1μmol/L MNNG and cultured for 40 passages. During the culturing process, cell proliferation, migration, and soft agar colony formation significantly increased after 30 passages. Following MNNG exposure, lactate, LDH, glucose uptake, and the expression levels of key enzymes in glycolysis were significantly increased. Knocking down LDHA or LDHB alone reduced lactate secretion, inhibited cell viability, and impaired migratory capacities. Knocking down LDHA and LDHB together fully suppressed lactate secretion and effectively suppressed the malignant phenotype of cells transformed by long-term MNNG exposure. Finally, we demonstrated that overexpression of LDHA and LDHB promotes the malignant transformation of GES-1cells by enhancing the Warburg effect during long-term exposure to NOCs.