Long-term Supplementation of Deep-fried Oil Consumption Impairs Oxidative Stress, Colon Histology and Increases Neurodegeneration.

BACKGROUND: Sesame oil and sunflower oil are popular cooking oils in southern India. Deep-frying is a frequent method of food preparation. Deep-frying at high temperatures has been linked with several disorders, including cancer, diabetes, and unknown metabolic problems. There have been no long-term investigations on the influence of deep-fried oils on PUFA metabolism and pathogenesis. As a result, the current study aimed to explore the effect of deep-fried frying oil on Wistar rats by continuous treatment. Furthermore, the pathophysiology of MSG-induced neurotoxicity in Wistar rats was investigated. METHODS: Wistar rats weighing 200-260 g were used in this study. Female rats were divided into five groups fed with (1) standard chow (control group), (2) unheated sesame oil (UHSO) along with standard chow, and (3) reheated sesame oil (RHSO) along with standard chow, (4) unheated sunflower oil (UHSFO) along with standard chow, and (5) reheated sunflower oil (RHSFO) along with standard chow and continued up to F1 generation. Furthermore, F1 male rats were treated with MSG of 2 g/kg body weight for 10 alternative days and were sacrificed for major tissues. RESULTS: We found that rats treated with RHSO and RHSFO showed increased body weight. Deep-fried oil-fed rats (RHSO and RHSFO) showed a significant increase in total cholesterol- 100 mg/dl, LDL- 23 mg/dl, & TAG-100 mg/dl, when compared to unheated oil rats. Liver function tests revealed that AST and ALT levels were significantly elevated in RHSO and RHSFO when compared to unheated oils and the control group. Inflammatory markers revealed that Hs-CRP (0.35 mg/dl) and LDH levels (6000 U/L) were significantly elevated in RHSO and RHSFO when compared to the unheated oils and control group. RT-PCR results showed significant elevation in the antioxidant genes SOD (twofold) and GPX (3-fold) when compared to UHSO and UHSFO groups. Liver and colon histology showed significant damage in the cell structure of RHSO and RHSFO-treated rats. Further, rats treated with unheated oils and MSG showed statistically significantly higher mRNA expression of neuroplasticity genes CREB, BDNF and reduced NMDA levels (UHSO, UHSFO) when compared to reheated oil groups (RHSO & RHSFO). Proinflammatory marker TNF-α expression was significantly elevated in RHSFO-treated rats when compared to control. Brain histology showed focal damage in glial cell degeneration in rats treated with RHSO and RHSFO when compared to other groups. CONCLUSION: The results from the present study proved that continuous supplementation deep-fried reheated oil consumption increased serum TGL and oxidative stress markers. Impaired liver metabolism and the involvement of the gut-liver-brain axis increased the risk of neurodegeneration.

Investigating the Association Between Fusobacterium nucleatum and Oral Squamous Cell Carcinoma: A Pilot Case-Control Study on Tissue Samples.

Background Fusobacterium nucleatum (F. nucleatum) has been increasingly linked to oral squamous cell carcinoma (OSCC), prompting this study to explore its presence using polymerase chain reaction (PCR) and evaluate its clinical significance. Methods In this pilot case-control study, 12 OSCC tissue samples and 12 non-cancerous oral mucosal tissue samples were analyzed. Total RNA extraction and complementary DNA (cDNA) synthesis were performed using Trizol-based methods, followed by PCR amplification and gel electrophoresis. The clinical characteristics of participants and PCR results were recorded. Results Among the OSCC tissue samples, three out of 12 tested positive for F. nucleatum, while none of the control samples showed its presence. The detection rate of F. nucleatum in OSCC was 25%. Gel analysis confirmed specific amplicon amplification, and ImageJ software enabled copy number quantification. Discussion Our findings support previous research indicating a potential association between F. nucleatum and OSCC. Understanding the etiological significance of F. nucleatum in OSCC has clinical implications, including early detection, risk stratification, and prognostication. However, the limited sample size and the need for further research to elucidate underlying mechanisms are acknowledged. Conclusion This pilot study provides initial evidence of F. nucleatum's presence in a subset of OSCC samples, supporting its potential association with oral cancer. Detecting F. nucleatum in OSCC tissues holds promise for future research and clinical applications as a diagnostic and prognostic biomarker. Understanding its role in oral carcinogenesis will facilitate the development of targeted therapeutic strategies. Larger studies are warranted to validate these findings and investigate the precise mechanisms involved.

New insights into the antibacterial mode of action of quercetin against uropathogen Serratia marcescens in-vivo and in-vitro.

In the course of a quest for therapeutic agents inhibiting uropathogens, the rise and universal blowout of antibiotic-resistant organisms is a wide problem. To overcome this matter, exploration of alternative antimicrobials is necessary. The antimicrobial potential of quercetin has been widely described against some pathogenic microorganisms, but to the best of our knowledge, no report exists against the pathogenicity of uropathogenic Serratia marcescens. Hence, the present study focused on the antibacterial mechanism of action of quercetin, a flavonoid against the uropathogen Serratia marcescens. Quercetin was evaluated for its anti-QS activity, and the attained outcomes showed that quercetin inhibited QS-mediated virulence factors such as biofilm formation, exopolysaccharides, swarming motility and prodigiosin in Serratia marcescens. The proposed mechanism of action of quercetin greatly influences cell metabolism and extracellular polysaccharide synthesis and damages the cell membrane, as revealed through global metabolome profiling. In vivo experiments revealed that treatment with quercetin prolonged the life expectancy of infected Caenorhabditis elegans and reduced the colonization of Serratia marcescens. Hence, the current study reveals the use of quercetin as a probable substitute for traditional antibiotics in the treatment of uropathogen infections driven by biofilms.

Maternal supplementation of high-value PUFA-Rich Isochrysis sp. biomass prevents monosodium glutamate-induced neurotoxicity in first-generation Wistar rats.

Prenatal supplementation of high-value PUFA (HVPUFA) is essential for adequate brain development in infants. As marine microalgal derived omega-3 fatty acids are considered an alternative source of fish oil, their neuroprotective role on monosodium glutamate (MSG)-induced neurotoxicity, bioavailability, and disease prevention in first-generation (F1) animals need to be explored at molecular level. This study tested the long term supplementation of microalgal derived ω-3 PUFAs from parent rats to its offspring rats and studied the neuroprotective role in monosodium glutamate (MSG)-induced neurotoxicity in F1 rats. The parent animals were divided into three groups: control, microalgal-administered group (5.7 mg of EPA and 1.4 mg of DHA/kg BW from Isochrysis sp.), and fish oil-administered group (4.2 mg of EPA and 2.9 mg of DHA/kg BW derived from fish oil) (FG) and continued up to F1 generation. The F1 male rats from respective parents were separated for disease induction: group I animals (control) were administered with 500 µl of Milli-q water alone and group II (disease control), III (Microalga), and IV (fish oil) animals were administered with 2 g/kg bodyweight of MSG for 10 alternative days. Microalga-treated F1 rats showed significant HDL (43 mg/dl) levels when compared to their experimental groups. Brain tissues of microalga-treated F1 rats (MG) showed higher concentration of DHA (10.1 mg/100 mg tissue) and ARA (18.7 mg/100 mg tissue) levels and significant reduction of MDA (30 nM mg protein) levels. Furthermore, MSG induced neurotoxicity was ameliorated through the activation of CREB and BDNF genes The mRNA expressions of CREB and BDNF were 1.5-fold higher and NMDA levels were 2.0-fold higher in treated groups compared to disease control group. However, the expressions of antioxidant genes (SOD, catalase, and GPX) and apoptotic genes (Bcl-2 and Caspase-3) were significantly reduced in MG treated F1 rats when compared to disease control rats. Histopathological results also showed minimal focal damage in the tissues of MG F1 rats. Prenatal and continuous supply of microalgal biomass improves brain DHA and greatly reduced the consequences of MSG neurotoxicity in F1 rats.

Anti-listerial activity of microalgal fatty acid methyl esters and their possible applications as chicken marinade.

Fatty acid methyl esters (FAMEs) from marine microalgae have been reported to possess antimicrobial activities against several Gram positive and Gram negative bacteria, but a majority of them needs to be explored. The objective of this study was to investigate the antibacterial activity, mechanism of FAMEs from selected marine microalgae against Listeria monocytogenes, and to elucidate its efficacy in food model. The minimum inhibitory concentration of FAMEs was calculated to be 155 µg/mL for Chromulina sp. and 162 µg/mL for Nannochloropsis sp. against L. monocytogenes. Time-killing kinetics showed that FAMEs efficiently inhibited the growth of L. monocytogenes in a time and concentration dependent manner. The mechanism of action of FAMEs was studied by analysing its effects at a MIC on the cellular metabolism, membrane permeability, and membrane integrity of L. monocytogenes. Transmission Electron Microscopy (TEM) results showed that cells exposed to FAMEs showed damaged cell membrane structure with leakage of the internal contents in the cells of L. monocytogenes. Fluorescence microscopy images showed that L. monocytogenes cells treated with FAMEs showed high dead cell population corresponding with propidium iodide positive cells. Furthermore, FAMEs significantly down regulated quorum sensing and biofilm related genes (DegU, FlaE, and FlaD). In vivo therapeutic potential of FAMEs revealed improved Caenorhabditis elegans survival and reduced intestinal colonization during L. monocytogenes infection. Growth of listeria was abolished in chicken meat during the cold storage of 9 days when the samples were pre-treated with FAMEs. These results suggest anti-L. monocytogenes activity of FAMEs and elucidated its use in food control of chicken meat at refrigerated conditions.

Structured form of DHA prevents neurodegenerative disorders: A better insight into the pathophysiology and the mechanism of DHA transport to the brain.

Docosahexaenoic acid (DHA) is one of the most important fatty acids that plays a critical role in maintaining proper brain function and cognitive development. Deficiency of DHA leads to several neurodegenerative disorders and, therefore, dietary supplementations of these fatty acids are essential to maintain cognitive health. However, the complete picture of how DHA is incorporated into the brain is yet to be explored. In general, the de novo synthesis of DHA is poor, and targeting the brain with specific phospholipid carriers provides novel insights into the process of reduction of disease progression. Recent studies have suggested that compared to triacylglycerol form of DHA, esterified form of DHA (i.e., lysophosphatidylcholine [lysoPC]) is better incorporated into the brain. Free DHA is transported across the outer membrane leaflet of the blood-brain barrier via APOE4 receptors, whereas DHA-lysoPC is transported across the inner membrane leaflet of the blood-brain barrier via a specific protein called Mfsd2a. Dietary supplementation of this lysoPC specific form of DHA is a novel therapy and is used to decrease the risk of various neurodegenerative disorders. Currently, structured glycerides of DHA - novel nutraceutical agents - are being widely used for the prevention and treatment of various neurological diseases. However, it is important to fully understand their metabolic regulation and mechanism of transportation to the brain. This article comprehensively reviews various studies that have evaluated the bioavailability of DHA, mechanisms of DHA transport, and role of DHA in preventing neurodegenerative disorders, which provides better insight into the pathophysiology of these disorders and use of structured DHA in improving neurological health.

Lowering the culture medium temperature improves the omega-3 fatty acid production in marine microalga Isochrysis sp. CASA CC 101.

Marine microalga Isochrysis sp. contains omega-3 fatty acids like eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Environmental factors play a major role in PUFA biosynthesis. Hence, the study focused to optimize factors such as temperature, pH, and photoperiod by response surface methodology (RSM). RSM results showed that the model is significant (p ≤ 0.05) with a high correlation coefficient (R2 = 0.908). The optimum conditions showed that maximum biomass (327 mg/L) at the temperature of 30 °C, pH of 7.5 and 16:8 (Light: Dark cycle), whereas the higher amount of DHA (13.3%) and EPA (9.0%) was observed in the conditions of 18 °C, pH of 7.5 and 16:8 (Light: Dark cycle). The biomass content was directly proportional to the temperature whereas DHA content was inversely proportional. It was revealed that the mRNA expression of EPA and DHA specific desaturases (5Des & 4Des) were significantly elevated in low temperature (20 °C) conditions. The results were highly correlated with the fatty acid profile of Isochrysis sp. grown under low temperature (20 °C) conditions which enhanced the EPA and DHA levels. This study suggests that the temperature is the most influencing factor which can be exploited in the industrial application of DHA and EPA production from Isochrysis sp.

Listeria monocytogens - Amended understanding of its pathogenesis with a complete picture of its membrane vesicles, quorum sensing, biofilm and invasion.

Listeria monocytogenes is a ubiquitous, intracellular foodborne pathogen that causes listeriosis in animals and humans. Pathogenic Listeria monocytogenes easily adapted to the conditions of human gastrointestinal tract and tolerate the counter changes such as acidity, bile, osmolarity, and antimicrobial peptides. They secrete specialized biologically active extra organ called membrane vesicles which comprises proteins, lipids, and lipopolysaccharides. Listerial vesicles possess functional versatility and play a significant role in pathogenesis by cell-free intercellular communication and toxin packaging. L. monocytogenes can attach promptly and decisively to inert substratum including intestinal mucosa, and forms biofilms and causes detrimental effects. Further, they invade the host cells through quorum sensing (QS) controlled virulence determinants and biofilms. The precise degree to which the bacterium retains the intracellular ambiance of host cells remains unknown. The machinery associated with intracellular survival, and the role of membrane vesicles, quorum sensing, and the Agr system in Listeria monocytogenes largely remains unclear. The current review focused to understand the role of membrane vesicles mediated pathogenesis biofilms, and delivers auxiliary impetus to understanding the potentials of virulence mediated invasion in Listeria monocytogenes.

Omega-3-rich Isochrysis sp. biomass enhances brain docosahexaenoic acid levels and improves serum lipid profile and antioxidant status in Wistar rats.

BACKGROUND: Isochrysis sp. is a marine microalga, rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The potential use of its biomass as an alternative source of polyunsaturated fatty acids (PUFAs) has not been studied in animal models. Male albino Wistar rats were divided into three groups and treated for 28 days. The rats were fed with (1) standard chow (control group), (2) microalgal biomass rich in EPA and DHA along with standard chow (microalga group), and (3) fish oil that contains equivalent amounts of EPA and DHA along with standard chow (fish oil group). After intervention, biochemical indices, histopathological indices, relative mRNA expression of PUFA genes, antioxidant genes, inflammatory markers, and the fatty acid profile of major tissues were studied. RESULTS: Animals treated with microalgal biomass showed significantly increased serum HDL levels (P < 0.05) and reduced oxidative stress markers with a concomitant decrease in urea and creatinine levels. Oral supplementation of microalgal biomass did not show any toxicity or damage in any major organs. The mRNA expression of PUFA genes was significantly downregulated (P < 0.05) and antioxidant genes were upregulated. Furthermore, the mRNA expression of pro-inflammatory markers was significantly downregulated (P < 0.05) and anti-inflammatory markers were upregulated. Oral supplementation of microalgal biomass improved DHA status in brain and liver. CONCLUSION: The present study demonstrated that Isochrysis sp. can be used as a safe, alternative food supplement for ω-3 fatty acids. © 2019 Society of Chemical Industry.