Harnessing the potential of microalgae for the production of monoclonal antibodies and other recombinant proteins.

Monoclonal antibodies (mAbs) have become indispensable tools in various fields, from research to therapeutics, diagnostics, and industries. However, their production, primarily in mammalian cell culture systems, is cost-intensive and resource-demanding. Microalgae, diverse photosynthetic microorganisms, are gaining attention as a favorable option for manufacturing mAbs and various other recombinant proteins. This review explores the potential of microalgae as a robust expression system for biomanufacturing high-value proteins. It also highlights the diversity of microalgae species suitable for recombinant protein. Nuclear and chloroplast genomes of some microalgae have been engineered to express mAbs and other valuable proteins. Codon optimization, vector construction, and other genetic engineering techniques have significantly improved recombinant protein expression in microalgae. These accomplishments demonstrate the potential of microalgae for biopharmaceutical manufacturing. Microalgal biotechnology holds promise for revolutionizing the production of mAbs and other therapeutic proteins, offering a sustainable and cost-effective solution to address critical healthcare needs.

2-Heptanone, 2-nonanone, and 2-undecanone confer oxidation off-flavor in cow milk storage.

Flavor sensation is one of the most prevalent characteristics of food industries and an important consumer preference regulator of dairy products. So far, many volatile compounds have been identified, and their molecular mechanisms conferring overall flavor formation have been reported extensively. However, little is known about the critical flavor compound of a specific sensory experience in terms of oxidized off-flavor perception. Therefore, the present study aimed to compare the variation in sensory qualities and volatile flavors in full-fat UHT milk (FFM) and low-fat UHT milk (LFM) samples under different natural storage conditions (0, 4, 18, 25, 30, or 37°C for 15 and 30 d) and determine the main component causing flavor deterioration in the FFM and LFM samples using sensory evaluation, electronic nose, and headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). In addition, the Pearson correlation between the volatile flavor components and oxidative off-flavors was analyzed and validated by sensory reconstitution studies. Compared with the LFM samples, the FFM samples showed a higher degree of quality deterioration with increased storage temperature. Methyl ketones of odd carbon chains (i.e., 2-heptanone, 2-nonanone, 2-undecanone, 2-tridecanone, and 2-pentadecanone) reached a maximum content in the FFM37 samples over 30 d storage. The combined results of the Pearson correlation and sensory recombination study indicated that 2-heptanone, 2-nonanone, and 2-undecanone conferred off-flavor perception. Overall, the present study results provide potential target components for detecting and developing high-quality dairy products and lay a foundation for specific sensory flavor compound exploration in the food industry.

Correlation between glycaemic state and tooth mobility in patients with periodontal disease.

OBJECTIVE: To evaluate the association of tooth mobility with glycaemic levels in patients with periodontitis. METHODS: The cross-sectional study was conducted at the Department of Oral Medicine, Ziauddin Dental Hospital, Karachi, from December 2018 to May 2019, and comprised patients of either gender with chronic periodontitis. After recording demographic details and dental charting, tooth mobility scores were correlated with gingival crevicular blood glucose, finger capillary blood glucose and glycosylated haemoglobin levels using Pearson's correlation. Linear regression was applied to assess the inter-relation between the variables. Data was analysed using SPSS 20. RESULTS: Of the 348 patients, 202(58%) were females and 146(42%) were males. The overall mean age was 43±10.4 years. The mean number of teeth in patients with glucose levels <180mg/dl was 25.5±2.5 compared to 23.2±2.9 in individuals with glucose levels >200mg/dl. A moderate positive correlation (r=0.658) was seen between gingival crevicular blood glucose levels and tooth mobility. Finger capillary blood glucose levels also showed good correlation (r=0.653) with tooth mobility scores. Glycosylated haemoglobin scores showed a strong positive correlation(r=0.733). Linear regression confirmed increased glycaemic levels as a risk factor for tooth mobility (p<0.001). CONCLUSIONS: Tooth mobility and glycaemic levels were found to be strongly interrelated.

Efflux Transporters' Engineering and Their Application in Microbial Production of Heterologous Metabolites.

Metabolic engineering of microbial hosts for the production of heterologous metabolites and biochemicals is an enabling technology to generate meaningful quantities of desired products that may be otherwise difficult to produce by traditional means. Heterologous metabolite production can be restricted by the accumulation of toxic products within the cell. Efflux transport proteins (transporters) provide a potential solution to facilitate the export of these products, mitigate toxic effects, and enhance production. Recent investigations using knockout lines, heterologous expression, and expression profiling of transporters have revealed candidates that can enhance the export of heterologous metabolites from microbial cell systems. Transporter engineering efforts have revealed that some exhibit flexible substrate specificity and may have broader application potentials. In this Review, the major superfamilies of efflux transporters, their mechanistic modes of action, selection of appropriate efflux transporters for desired compounds, and potential transporter engineering strategies are described for potential applications in enhancing engineered microbial metabolite production. Future studies in substrate recognition, heterologous expression, and combinatorial engineering of efflux transporters will assist efforts to enhance heterologous metabolite production in microbial hosts.

Ultrasonic pretreatment improved the antioxidant potential of enzymatic protein hydrolysates from highland barley brewer's spent grain (BSG).

Highland barley brewer's spent grain (BSG), being China's brewing industry's major by-product is the focus of current research. The aim of the present study was to scrutinize the effects of ultrasound and heat pretreatments on enzymatic hydrolysis of highland barley BSG protein hydrolysates (HBSGPH) and evaluate the effect of enzymatic hydrolysis time on the antioxidant activity of hydrolysates by Alcalase. Different ultrasonic waves (40 and 50 kHz) and heat pretreatment temperatures (50 and 100 °C) were chosen and the pretreatment time was 15, 30, and 60 min. The obtained results revealed that the ultrasound pretreatment of highland barley BSG protein at 40 and 50 kHz has significantly (P < 0.05) enhanced about 57 and 67% of oxygen radical absorption capacity of obtained hydrolysate over the untreated substrate. The 1,1-diphenyl-2-picrylhdrazl (DPPH) radical scavenging activity (DRSA) 28%, metal chelating activity (MCA) 54%, superoxide radical scavenging activity (SRSA) 18%, and hydroxyl radical scavenging activity (HRSA) 25% of HBSGPH at 50 kHz were also improved (P < 0.05) significantly. HBSGPH from heat treatment at 100 °C showed no SRSA and HRSA scavenging activities but improved significantly (P < 0.05) about 27% ferric reducing antioxidant power (FRAP) assay values. In the present work, the resultant HBSGPH had stronger antioxidant properties with ultrasound pretreatment at 50 kHz and the enzymatic hydrolysis after 4 hr was facilitating the enzymatic release of antioxidant peptides from HBSGPH. PRACTICAL APPLICATION: Highland barley BSG is attracting toward natural food products due to its potent natural antioxidants to overcome the risk of diseases and are beneficial for human health.

Clinical efficacy of photodynamic therapy as an adjunct to scaling and root planing in the treatment of chronic periodontitis among cigarette smokers: A systematic review and meta-analysis.

BACKGROUND: The aim of the study was to evaluate the clinical efficacy of adjunctive photodynamic therapy (PDT) as compared to scaling and root planing (SRP) alone in the treatment of chronic periodontitis among cigarette smokers. METHODS: Electronic databases including EMBASE, MEDLINE, Cochrane Oral Health Group Trials Register and Cochrane Central Register of Controlled Trials were searched up to January 2019. Randomized clinical trials (RCTs) with data on comparison between adjunctive PDT compared to SRP in each group were included. Primary outcomes included clinical attachment level (CAL) gain, while secondary outcomes was probing depth (PD) reduction. The weighted mean differences (WMD) of outcomes and 95% confidence intervals (CI) for each variable were calculated using random and fixed effect models. RESULTS: A total of 5 RCTs were included. Two clinical trials were classified as having a low risk of bias and 3 clinical trials as high risk of bias. The overall effect for CAL gain (WMD= -0.088, 95% CI= -0.40 to 0.23, p = 0.58) and PD reduction (WMD= -0.35, 95% CI= -0.87 to -0.17, p = 0.18) was not statistically significant between PDT and SRP groups at follow-up, respectively. CONCLUSION: Within the limitations of this study, it remains debatable whether PDT as an adjunct to SRP is more effective in clinical attachment level gain as compared to SRP alone in cigarette smokers given that the available scientific evidence is weak.

Cytokine levels and their role in the etiopathogenesis of Burning Mouth Syndrome: A systematic review.

INTRODUCTION: Burning Mouth Syndrome is characterized by variable symptoms that include pain, burning and paraguesia in an otherwise healthy-appearing oral mucosa. Although the etiopathogenesis of Burning Mouth Syndrome is unknown, some studies provide evidence of subclinical inflammation leading to disrupted cytokine levels. AIM: To investigate the expression of cytokines and role in the etiopathogenesis of Burning Mouth Syndrome. METHODS: Online databases (MEDLINE and EMBASE) were searched from November 1986 to November 2018 for case control/cross-sectional studies comparing the levels of cytokines in patients with Burning Mouth Syndrome and healthy controls. RESULTS: A total of eight studies were included in the current review. Four studies were of high and four studies were of moderate quality. Seven studies evaluated IL-6, out of which four showed comparable results, two showed higher levels and one study reported lower levels in Burning Mouth Syndrome patients compared to controls. Four studies assessed IL-2, out of which two reported comparable results whereas one study reported higher levels and one study reported lower levels in Burning Mouth Syndrome patients compared to controls. IL-10 levels were measured in three studies that reported no significant differences in the levels between Burning Mouth Syndrome and healthy controls. DISCUSSION AND CONCLUSION: The etiopathogenesis of Burning Mouth Syndrome is multifactorial. Studies have provided scientific evidence that inflammation plays a key role in Burning Mouth Syndrome pathogenesis. However, whether up-regulation or down-regulation of specific cytokines contribute to the etiopathogenesis of Burning Mouth Syndrome remains debatable. Further high-quality studies with larger sample size and assessing a wider array of cytokines are warranted in order to obtain strong conclusions.

Differential expression of salivary S100A7 in oral submucous fibrosis.

AIM: To investigate the expression of salivary S100A7 levels among patients with oral submucous fibrosis (OSF) and healthy controls. METHOD: A total number of 60 participants were included in the study (30 OSF cases and 30 healthy controls). Demographic data was collected using a structured baseline questionnaire. Salivary S100A7 levels were quantified using enzyme-linked immunosorbent assay. Data was analyzed using Student t-test. Pearson correlation test was used to evaluate correlation between S100A7 levels and independent variables such as frequency and duration of areca nut use, gutka use, and mouth opening. RESULTS: The mean value of salivary S100A7 for OSF group was 0.275 ng/ml, whereas mean value of salivary S100A7 for healthy controls was 0.195 ng/ml. Student t-test indicated that there was statistically significantly higher levels of S100A7 in OSF group as compared to healthy controls (p < .001). When the clinical variables of individual groups were analysed, a significant negative correlation was found between salivary S100A7 and duration of areca nut (p = .009) and gutka chewing (p = .03), whereas a significant positive correlation was found for mouth opening (p = .04). CONCLUSION: OSF presented higher levels of salivary S100A7 levels as compared with healthy individuals and may be used as surrogate measure to identify subjects at risk for OSF.

Effect of local probiotic (Lactobacillus reuteri) vs systemic antibiotic therapy as an adjunct to non-surgical periodontal treatment in chronic periodontitis.

AIM: The aim of the present study was to assess and compare the clinical efficacy of local probiotic Lactobacillus reuteri (L. reuteri) and systemic antibiotics as adjunct to scaling and root planing (SRP) in the treatment of chronic periodontitis (CP). METHODS: Thirty systemically-healthy participants (15 probiotic and 15 antibiotic), clinically diagnosed with CP, were enrolled. All patients underwent SRP. Adjunctive probiotics were administered twice daily for 3 months, whereas a combination of amoxicillin and metronidazole were given three times daily for 7 days. RESULTS: Intragroup analysis showed statistically-significant improvement in all clinical parameters: plaque index, bleeding on probing, periodontal pocket depth, and clinical attachment level gain at each follow-up visit. However, intergroup comparison of clinical periodontal parameters did not show statistical significance. CONCLUSION: The adjunctive use of L. reuteri and systemic antibiotics along with SRP showed similar improvement in all clinical periodontal parameters. This indicates that both adjunctive therapeutic agents showed similar efficacy in resolving inflammation and improving periodontal outcomes.

Systematic review and meta-analysis of double-blind, placebo-controlled, randomized clinical trials using probiotics in chronic periodontitis.

AIM: The aim of the present study was to evaluate the efficacy of probiotics as an adjunct to scaling and root planning (SRP) in the treatment of chronic periodontitis (CP). METHODS: The focused question of the study was: Does adjunctive use of probiotics yield better clinical periodontal outcomes compared to placebo/no treatment group in the treatment of CP? Electronic and manual literature searches were conducted up to December 2017 using the following databases: MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, and Cochrane Oral Health Group Trials Register. Forest plots were computed reporting weighted mean difference (WMD) of outcomes and 95% confidence intervals (CI). RESULTS: Seven clinical studies were included. Four studies showed additional benefits in reducing periodontal probing depth (PPD) and gaining clinical attachment level (CAL), whereas, three studies showed comparable clinical periodontal outcomes between probiotics and SRP/placebo. Significant heterogeneity was observed for PPD reduction and CAL gain. The overall mean difference for CAL gain between probiotics and placebo/SRP was significant (WMD = 1.41, 95% CI = 0.15-2.67, P = .028) at follow up. CONCLUSION: Adjunctive probiotics could result in additional benefits in CAL gain in CP. Nevertheless, further high-quality randomized clinical trials with microbiological outcomes are warranted to obtain strong conclusions in this regard.

S100 proteins in oral squamous cell carcinoma.

BACKGROUND: Controversy exists in the literature regarding the differential expression of S100 protein members and their functional correlations in oral squamous cell carcinoma (OSCC). The aim of the present study was to systematically review the expression of S100 protein family members among OSCC and healthy controls and to evaluate whether S100 protein members serve as diagnostic marker in OSCC. METHODS: Indexed databases were searched up to and including October 2017. Case-control/cross-sectional studies in human diagnosed clinically and/or histologically with OSCC and evaluated the expression of S100 protein family among OSCC and healthy controls were included. RESULTS: A total of 11 studies were included. Four studies were of good quality, 5 were of moderate and 2 were of poor quality. Five studies evaluated S100A2, A7 and A12 and showed overexpression of these protein levels in OSCC patients when compared to healthy controls. Three studies reported down-regulation of S100A1, A3, A6, A11, A13, A14, A16 and S100Z in OSCC patients as compared to healthy controls. Two studies reported overexpression of S100A9 and one study each reported overexpression of S100A4, A8, A10, and S100P in OSCCs as compared to healthy controls respectively. CONCLUSION: It remains debatable whether up-regulation or down-regulation of specific S100 protein members serves as a diagnostic marker in OSCC. With the findings of the present systematic review, the threshold for diagnostic levels of S100 proteins cannot be proposed. In addition, S100A7 protein could act as a potential OSCC marker. However, further case-control studies with larger sample size are required to obtain strong conclusion in this regard.

Hutchinson-Gilford Progeria Syndrome: A Premature Aging Disease.

Progeria is sporadic, very rare, autosomal dominant, deadly childhood disorder. It is one of the progeroid syndromes also known as Hutchinson-Gilford progeria syndrome (HGPS). Aging is a developmental process that begins with fertilization and ends up with death involving a lot of environmental and genetic factors. The disease firstly involves premature aging and then death from complications of atherosclerosis such as myocardial infarction, stroke, atherosclerosis, or heart failure. The lifespan of the patient is normally up to teen age or early twenties. It is usually not inherited because a patient normally dies before the age of reproduction. The most important genetic linkage between progeria and aging is shortening of telomere ends with each replication cycle. The patients are normally observed to have extremely short telomeres. Currently, 90% of the patients are said to have de novo point mutations in the LMNA gene that substitute cytosine with thymine and have been found in individuals with HGPS. Lmna encodes lamins A and C, and the A-type lamins have important structural function in the nuclear envelope. The most common type of HGPS mutation is located at codon 608 (G608G). It could not be diagnosed at birth, but after the age of 2 years, visible, prominent symptoms can be observed. Still, lot of research is needed to solve this mystery; hopefully, future research on HGPS would provide important clues for progeria and other fatal age-related disorders.

Composition and Nutrient Value Proposition of Brewers Spent Grain.

Brewer's spent grain (BSG), a major brewing industry byproduct, is generated in large quantities annually. This review summarizes research into the composition and preservation of BSG, different extraction techniques for BSG proteins and phenolic acids, and the bioactivities of these phenolic components. Moreover, this article also highlights BSG integration into foodstuff for human consumption and animal feed supplements. BSG is considered a rich source of fiber, protein, and phenolic compounds. The phenolic acids present in BSG are hydroxycinnamic acids (ferulic, p-coumaric, and caffeic acids), which have many biofunctions, such as antioxidant, anticarcinogenic, antiatherogenic, and antiinflammatory activities. Previously, attempts have been made to integrate BSG into human food, such as ready-to-eat snacks, cookies and bread, to increase fiber and protein contents. The addition of BSG to animal feed leads to increased milk yields, higher fat contents in milk, and is a good source of essential amino acids. Therefore, many studies have concluded that integrating the biofunctional compounds in BSG into human food and animal feed has various health benefits.