Phlorotannins-bioactivity and extraction perspectives.

Phlorotannins, a seaweed based class of polyphenolic compounds, have proven to possess potential bioactivities such as antioxidant, antimicrobial, anti-allergic, anti-diabetic, anti-inflammatory, anti-cancerous, neuroprotection etc. These bioactivities have further increased demand globally and sustainable techniques such as supercritical fluid extraction, microwave assisted extraction, enzyme assisted extraction, extraction using deep eutectic solvents etc. are being explored currently for production of phlorotannin-rich extracts. In spite of such well documented bioactivities, very few phlorotannin-based nutraceuticals are available commercially which highlights the significance of generating consumer awareness about their physiological benefits. However, for industry level commercialization accurate quantification of phlorotannins with respect to the different classes is vital requiring sophisticated analytical techniques such as mass spectrometry, 1H-NMR spectroscopy etc. owing to the wide structural diversity. This review summarizes the extraction and bioactivities of phlorotannins based on the findings of in vivo and in vitro studies.

Exploring the physical and quality attributes of muffins incorporated with microencapsulated squalene as a functional food additive.

Squalene, a triterpenoid compound is proven to possess immense bioactivities by virtue of its high antioxidant activity. The present study was designed to investigate the quality attributes of muffins as influenced by addition of encapsulated squalene. Nutritional analysis showed that calorific value of prepared muffins has ranged from 480.78 ± 0.10 to 501.61 ± 0.38 kcal. Baking loss was lowest in case of muffins prepared with encapsulated squalene with its crumb region recorded higher moisture content. Color kinetics study indicated that browning index (BI) was higher in crust portion of encapsulated squalene enriched muffins. Scanning electron micrographs showing that muffins with encapsulated squalene had stronger structural organization. This was further supported by the textural studies showed that the muffins with encapsulated squalene was cohesive, springier and chewy with less gumminess and stiffness indicating their efficacy in improving the textural quality. Oxidative stability and microbiological quality were also high in squalene enriched foods suggesting that squalene might have some antimicrobial effects. Outcome of the study indicated that encapsulated squalene can be very well utilised as a functional food ingredient in ready -to-eat functional foods. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s13197-020-04955-9).

Inhibition of growth and virulence of Vibrio cholerae by carvacrol, an essential oil component of Origanum spp.

AIMS: In the age where bacterial resistance to conventional antibiotics is increasing at an alarming rate, the use of the traditional plant, herb extracts or other bioactive constituents is gradually becoming popular as an anti-virulence agent to treat pathogenic diseases. Carvacrol, a major essential oil fraction of Oregano, possesses a wide range of bioactivities. Therefore, we aimed to study the effect of sub-inhibitory concentrations of carvacrol on major virulence traits of Vibrio cholerae. METHODS AND RESULTS: We have used in vitro as well as ex vivo models to access the anti-pathogenic role of carvacrol. We found that the sub-inhibitory concentration of carvacrol significantly repressed bacterial mucin penetrating ability. Carvacrol also reduced the adherence and fluid accumulation in the rabbit ileal loop model. Reduction in virulence is associated with the downregulated expression of tcpA, ctxB, hlyA and toxT. Furthermore, carvacrol inhibits flagellar synthesis by downregulating the expression of flrC and most of the class III genes. CONCLUSIONS: Carvacrol exhibited anti-virulence activity against V. cholerae, which involved many events including the inhibition of mucin penetration, adhesion, reduced expression of virulence-associated genes culminating in reduced fluid accumulation. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings indicate that carvacrol possesses inhibitory activity against V. cholerae pathogenesis and might be considered as a potential bio-active therapeutic alternative to combat cholera.

Chitosan - Whey protein as efficient delivery system for squalene: Characterization and functional food application.

Squalene, a triterpenoid compound possessing excellent bioactivities, is not being utilized as a functional food ingredient due to its high susceptibility to oxidation. In the present study, the feasibility of chitosan-whey protein as an efficient wall material for squalene encapsulation using spray drying technique was attempted for functional food applications. The encapsulation efficiency of the squalene powder was found to be 75.4 ±â€¯0.22% whereas other physico-chemical properties such as moisture content, flowability, solubility, peroxide value, etc. have shown satisfactory results. The thermogravimetric analysis showed that chitosan-whey protein was able to retain the thermal stability of squalene up to a temperature of 422 °C. Furthermore, the functional food application of the encapsulated squalene in a bakery product (cake) exhibited significantly (p < 0.05) better properties in terms of oxidative stability, sensory attributes than that of cake with pure squalene and control treatment. Hence, it can be concluded that emulsification of squalene in chitosan-whey protein and its subsequent encapsulation by spray drying can be a potential process to produce oxidatively stable encapsulates for the development of functional foods.

Identification of common virulence factors present in enterotoxigenic Escherichia coli isolated from diarrhoeal patients in Kolkata, India.

AIMS: Development of an effective vaccine against enterotoxigenic Escherichia coli (ETEC) is largely dependent on the conscientious understanding of different virulence associated factors from diverse geographical areas. So, the objective of this study is to elucidate the distribution of enterotoxins, CF and NCVF in clinical ETEC strains isolated between 2008 and 2014 from two hospitals in Kolkata, India. METHODS AND RESULTS: Multiplex PCR method was used for detection of two enterotoxin genes, 11 common CFs and five common NCVFs. Among the 350 tested ETEC strains, 61% strains possessed est+elt genes, 25% est and 14% elt. Among 56% CF positive ETEC strains, CS21 was the prevalent one (37%) followed by CS6 (36%). NCVF genes were present in 59% of the ETEC strains; eatA was the most prevalent (65%) followed by etpA (51%). There were 29% strains negative for any CFs or NCVFs. CONCLUSIONS: We conclude that a pattern exists between CS6, eatA and toxins. We observed est with or without elt, CS6 with or without CS5 and with or without eatA were present in 24% of clinical ETEC strains (59/250) analysed. CS21 has emerged as another predominant CF but it had diverse CFs and NCVFs. SIGNIFICANCE AND IMPACT OF THE STUDY: Prevalence of ETEC virulence factors would help in tracking ETEC globally and suggests the need of a multivalent ETEC vaccine.

Prediction of clinical outcome in patients treated with cardiac resynchronization therapy - the role of NT-ProBNP and a combined response score.

BACKGROUND: Cardiac resynchronization therapy (CRT) is an established therapy for appropriately selected patients with heart failure. Response to CRT has been heterogeneously defined using both clinical and echocardiographic measures, with poor correlation between the two. METHODS: The study cohort was comprised of 202 CRT-treated patients and CRT response was defined at 6 months post-implant. Echocardiographic response (E+) was defined as a reduction in LVESV ≥ 15%, clinical response as an improvement of ≥ 1 NYHA class (C+), and biomarker response as a ≥ 25% reduction in NT-proBNP(B+). The association of response measures (E+, B+, C+; response score range 0-3) and clinical endpoints at 3 years was assessed in landmarked Cox models. RESULTS: Echo and clinical responders demonstrated greater declines in NT-proBNP than non-responders (median [E+/B+]: -52%, [E+]: -27%, [C+]: -39% and [E-/C-]: -13%; p = 0.01 for trend). Biomarker (HR 0.43 [95% CI: 0.22-0.86], p = 0.02) and clinical (HR 0.40 [0.23-0.70] p = 0.001) response were associated with a significantly reduced risk of the primary endpoint. When integrating each response measure into a composite score, each 1 point increase was associated with a 31% decreased risk for a composite endpoint of mortality, LVAD, transplant and HF hospitalization (HR 0.69 [95% CI: 0.50-0.96], p = 0.03), and a 52% decreased risk of all-cause mortality (HR 0.48 [95% CI: 0.26-0.89], p = 0.02). CONCLUSION: Serial changes in NT-proBNP are associated with clinical outcomes following CRT implant. Integration of biomarker, clinical, and echocardiographic response may discriminate CRT responders versus non-responders in a clinically meaningful way, and with higher accuracy. TRIAL REGISTRATION: The cohort was combined from study NCT01949246 and the study based on local review board approval 2011/550 in Lund, Sweden.

Antioxidant, functional properties and amino acid composition of pepsin-derived protein hydrolysates from whole tilapia waste as influenced by pre-processing ice storage.

In recent years, hygienic handling of fishery waste is demanded owing to the fact that the fishery waste is an ideal raw material for the preparation of bioactive compounds. In the present study, the effect of pre-processing storage (at 4 ± 2 °C) of whole tilapia waste (WTW) on the properties of its protein hydrolysate derived using pepsin was evaluated. Fish protein hydrolysates (FPH) were prepared from 0, 24 and 48 h old ice stored WTW and designated as FPH-0, FPH-1, and FPH-2, respectively. Total amino acids, total essential amino acids and total hydrophobic amino acids of FPH samples increased with the storage period of raw material (WTW). Antioxidant activities such as DPPH (2, 2 diphynyl-1-picrylhydrazyl) free radical scavenging activity and ferric reducing power of FPH samples were dose dependent. FPH-0 had better antioxidant properties including linoleic acid peroxidation inhibition activity than FPH-1 and FPH-2. The DNA nicking assay revealed the protective effect of FPH preparations against Fenton's reaction mediated oxidative damage. FPH-2 had better emulsifying properties and foaming stability whereas the FPH-0 had relatively good foaming capacity. SDS-PAGE indicated the presence of peptides ranging from 116 to < 14.4 kDa in FPH-0 and less than 18 kDa in FPH-1 and FPH-2. The present study, clearly demonstrated that whole tilapia waste can effectively be converted to FPH and could be a potential ingredient in functional food and as a rich source of high-quality protein in animal feed formulations.

Evaluation of chitosan as a wall material for microencapsulation of squalene by spray drying: Characterization and oxidative stability studies.

The present study was aimed at investigating the efficacy of chitosan as a wall material for microencapsulation of squalene by spray drying for functional food applications. Based on different core to wall material ratio (1:1, 0.5:1 and 0.3:1 on w/w basis), emulsions were prepared and evaluated in terms of emulsion stability, particle size, zeta potential, polydispersity Index (PDI), rheology and microstructure. The optimized emulsion combination was spray dried and characterized, physically and chemically. The encapsulation efficiency of the powder was found to be 26±0.6% whereas other properties such as particle size, zeta potential, water activity, hygroscopicity, Carr Index, Hausner ratio have shown satisfactory results. SEM analysis showed that the squalene microcapsules were smooth spherical particles free from dents and fissures. FTIR data further confirmed the encapsulation of squalene with chitosan. However, TGA, oxidative stability and accelerated Rancimat results showed that chitosan was not able to protect squalene from oxidation during storage. The results suggest that chitosan is not an appropriate wall material for microencapsulation of squalene and hence a combination of wall materials could be attempted for the encapsulation of squalene.

Dietary supplementation of thiamine and pyridoxine-loaded vanillic acid-grafted chitosan microspheres enhances growth performance, metabolic and immune responses in experimental rats.

In the present investigation, the effect of dietary supplementation of thiamine and pyridoxine loaded vanillic acid-grafted chitosan microspheres (TPVGC) on growth, metabolic and immune responses in Wistar strain albino rats was studied. Eight experimental groups, namely four groups each for male and female rats were fed with 0, 0.4, 0.8 and 1.6% of TPVGC in the diet. At the end of 45days feeding trials, both male and female rats supplemented with TPVGC had higher weight gain% and specific growth rate than the control groups. Significantly (p<0.05) lower blood glucose level and higher respiratory burst activity were recorded in the treatment groups than the control groups of both male and female rats. Activity of metabolic enzymes (aspartate amino transferase, alanine aminotransferase, alkaline phosphatase and acid phosphatase) and antioxidant enzymes (superoxide dismutase, catalase and glutathione S-transferase) were significantly higher (p<0.05) in the control groups and a decreasing trend in the same was observed with a gradual increase in the inclusion level of TPVGC in the diet of the treatment groups. However, a reverse trend was observed for acetylcholine esterase. It was inferred that dietary supplementation of thiamine and pyridoxine loaded vanillic acid-grafted chitosan enhanced the growth performance, metabolic and immune responses in the animal-model.

Anandamide rescues retinal barrier properties in Müller glia through nitric oxide regulation.

The blood retinal barrier (BRB) can mitigate deleterious immune response. Dysfunction at the BRB can affect disease progression. Under inflammatory conditions Müller glia produce increased pro-inflammatory factors, like nitric oxide (NO). In this study we describe molecular events at the Müller glia during inflammation which could affect inner BRB properties. Griess assay and 4,5-diaminofluorescein diacetate (DAF-2DA) time-lapse fluorescence were used to measure NO production. Western blot was used to analyze the expression of inducible nitric oxide synthase (iNOS) and mitogen-activated protein kinases (MAPK) components. Lucifer Yellow was used to measure permeability. Griess assay and DAF-2DA time-lapse fluorescence images revealed that lipopolysaccharide (LPS) induced inflammation and increased NO production. In parallel, changes were observed in tight junction proteins, zona occludens 1 (ZO-1), connexin 43 (Cx43), and permeability. This was mediated through activation of iNOS and mitogen-activated protein kinase phosphatase-1 (MKP-1), implicated in immune response. Endocannabinoids can exert a protective and anti-inflammatory effect. Exogenous arachidonoyl ethanolamide (AEA) inhibited NO generation and also abolished LPS-induced increase in permeability. Our work suggests that subtle changes in Müller glia function, which act as part of the BRB, could contribute to retinal health. AEA which can reduce inflammatory cytotoxicity has potential as treatment in several ocular manifestations where the integrity of the BRB is crucial.

Ultrastructural alterations in the gills of Labeo rohita fingerlings exposed to thermal extremes.

This study aimed to determine the cellular alterations in the gill of Labeo rohita exposed to lethal temperature maxima (LTM ax ) and lethal temperature minima (LTM in ) by means of transmission electron microscopy (TEM). Acclimation of advanced fingerlings of L. rohita was carried out at 26°C for 30 days. Acclimated fish were subjected to a constant rate of increase or decrease in temperature (0.3°C/min) until the LTM ax and LTM in values were reached. Dissected gills were processed for TEM, both at the end of acclimation period at ambient temperature (26°C) and at lethal temperatures. Results indicated that at ambient temperature, the gill tissues appeared normal. However, significant changes were observed at lethal temperatures. The gill tissues at lethal temperature maxima showed severely damaged lamellae, with more vacuolated space. At lethal temperature minima, gill tissues showed increased density of mitochondria. Our prima-facie report indicated that L. rohita exposed to lethal temperatures exhibited marked ultrastructural changes in the gills.

Common genetic variants in the 9p21 region and their associations with multiple tumours.

BACKGROUND: The chromosome 9p21.3 region has been implicated in the pathogenesis of multiple cancers. METHODS: We systematically examined up to 203 tagging SNPs of 22 genes on 9p21.3 (19.9-32.8 Mb) in eight case-control studies: thyroid cancer, endometrial cancer (EC), renal cell carcinoma, colorectal cancer (CRC), colorectal adenoma (CA), oesophageal squamous cell carcinoma (ESCC), gastric cardia adenocarcinoma and osteosarcoma (OS). We used logistic regression to perform single SNP analyses for each study separately, adjusting for study-specific covariates. We combined SNP results across studies by fixed-effect meta-analyses and a newly developed subset-based statistical approach (ASSET). Gene-based P-values were obtained by the minP method using the Adaptive Rank Truncated Product program. We adjusted for multiple comparisons by Bonferroni correction. RESULTS: Rs3731239 in cyclin-dependent kinase inhibitors 2A (CDKN2A) was significantly associated with ESCC (P=7 × 10(-6)). The CDKN2A-ESCC association was further supported by gene-based analyses (Pgene=0.0001). In the meta-analyses by ASSET, four SNPs (rs3731239 in CDKN2A, rs615552 and rs573687 in CDKN2B and rs564398 in CDKN2BAS) showed significant associations with ESCC and EC (P<2.46 × 10(-4)). One SNP in MTAP (methylthioadenosine phosphorylase) (rs7023329) that was previously associated with melanoma and nevi in multiple genome-wide association studies was associated with CRC, CA and OS by ASSET (P=0.007). CONCLUSION: Our data indicate that genetic variants in CDKN2A, and possibly nearby genes, may be associated with ESCC and several other tumours, further highlighting the importance of 9p21.3 genetic variants in carcinogenesis.

Genome-wide association study meta-analysis of European and Asian-ancestry samples identifies three novel loci associated with bipolar disorder.

Meta-analyses of bipolar disorder (BD) genome-wide association studies (GWAS) have identified several genome-wide significant signals in European-ancestry samples, but so far account for little of the inherited risk. We performed a meta-analysis of ∼750,000 high-quality genetic markers on a combined sample of ∼14,000 subjects of European and Asian-ancestry (phase I). The most significant findings were further tested in an extended sample of ∼17,700 cases and controls (phase II). The results suggest novel association findings near the genes TRANK1 (LBA1), LMAN2L and PTGFR. In phase I, the most significant single nucleotide polymorphism (SNP), rs9834970 near TRANK1, was significant at the P=2.4 × 10(-11) level, with no heterogeneity. Supportive evidence for prior association findings near ANK3 and a locus on chromosome 3p21.1 was also observed. The phase II results were similar, although the heterogeneity test became significant for several SNPs. On the basis of these results and other established risk loci, we used the method developed by Park et al. to estimate the number, and the effect size distribution, of BD risk loci that could still be found by GWAS methods. We estimate that >63,000 case-control samples would be needed to identify the ∼105 BD risk loci discoverable by GWAS, and that these will together explain <6% of the inherited risk. These results support previous GWAS findings and identify three new candidate genes for BD. Further studies are needed to replicate these findings and may potentially lead to identification of functional variants. Sample size will remain a limiting factor in the discovery of common alleles associated with BD.

Aspirin but not ibuprofen use is associated with reduced risk of prostate cancer: a PLCO study.

BACKGROUND: Although most epidemiological studies suggest that non-steroidal anti-inflammatory drug use is inversely associated with prostate cancer risk, the magnitude and specificity of this association remain unclear. METHODS: We examined self-reported aspirin and ibuprofen use in relation to prostate cancer risk among 29 450 men ages 55-74 who were initially screened for prostate cancer from 1993 to 2001 in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. Men were followed from their first screening exam until 31 December 2009, during which 3575 cases of prostate cancer were identified. RESULTS: After adjusting for potential confounders, the hazard ratios (HRs) of prostate cancer associated with <1 and ≥ 1 pill of aspirin daily were 0.98 (95% confidence interval (CI), 0.90-1.07) and 0.92 (95% CI: 0.85-0.99), respectively, compared with never use (P for trend 0.04). The effect of taking at least one aspirin daily was more pronounced when restricting the analyses to men older than age 65 or men who had a history of cardiovascular-related diseases or arthritis (HR (95% CI); 0.87 (0.78-0.97), 0.89 (0.80-0.99), and 0.88 (0.78-1.00), respectively). The data did not support an association between ibuprofen use and prostate cancer risk. CONCLUSION: Daily aspirin use, but not ibuprofen use, was associated with lower risk of prostate cancer risk.

Use of external fixator in trochanteric fractures of femur.

External fixation of trochanteric fractures appears to be an effective option where major surgery is contra-indicated. A total of 26 cases of trochanteric fractures of femur (12 stable and 14 unstable) were treated with external fixation as other conventional treatment could not be applied due to associated comorbid factors related specially with the elderly age group and those unsuitable for major surgery and open trochanteric fractures. Ages of the patients varied from 38 to 93 (average being 64.8) years. Sixteen patients were female and 10 were male. Right side was affected in 12 and left side in 14 cases. All the cases were operated either by short general or spinal anaesthesia or even under local anaesthesia. All fractures united within 14-18 (average 16) weeks of time. According to Salvatti and Wilson scoring 10 patients showed good, 8 patients fair and 2 patients poor results. Out of other 6 patients, 2 patients died during course of treatment and 4 patients did not turn up for follow-up. Six patients suffered from superficial and two from deep pin tract infections. Other complications were noted such as bed sores in 2, mild to moderate degree of stiffness of both hips and knee joints in 2 and shortening of the affected limb (avarage 2 cm) in 17 cases. This procedure may be recommended to any patient with trochanteric fracture, who are unfit for major surgery and also for open fractures.

Efficacy of intra-articular dexmedetomidine for postoperative analgesia in arthroscopic knee surgery.

INTRODUCTION: Various drugs are administered intraarticularly to provide postoperative analgesia after arthroscopic knee surgery, of which opioids and alpha2 agonists could be of particular interest. METHODS: Sixty patients undergoing elective knee arthroscopy were randomly assigned to two groups (n = 30). Group R received 19 ml of 0.25% ropivacaine and 1 ml of isotonic saline (total volume 20 ml) intraarticularly. Group RD received 100 microg (1 ml) of dexmedetomidine added to 19 ml of 0.25% ropivacaine intra-articularly (total volume 20 ml). Analgesic effect was evaluated by measuring pain intensity (VAS score) and duration of analgesia. RESULTS: A longer delay was observed between intraarticular injection of study medication and first requirement of supplementary analgesic in group RD (10.84 +/- 2.6 hours) compared to group R (5.38 +/- 1.4 hours). Total consumption of fentanyl citrate in postoperative period was significantly less in group RD. No significant side effects were noted. CONCLUSION: Dexmedetomidine, added as adjunct to ropivacaine in patients undergoing arthroscopic knee surgery, improve the quality and duration of postoperative analgesia.