Comparison of Solvent-Free Microwave Extraction and Conventional Hydro-Distillation of Essential Oils from four Selected Underutilised Citrus sp. Plants

Essential oils (EOs) are widely used in cosmetic, pharmaceutical, fragrance and food industry. Solvent Free Microwave Extraction (SFME) is a green method for the extraction of essential oils from aromatic plants. The aim of the study is to compare SFME with conventional hydro-distillation (HD) technique, for the extraction of EO from four selected underutilised Citrus plant species in the essential oil industry; Citrus medica, C. reticulata, C. madurensis and C. limon in terms of chemical composition, antioxidant activity and energy consumption. The EOs were extracted using SFME (Microwave power: 700 W for 5 minutes and 400 W for 55 minutes) and HD (for 1 hour) and chemically analysed using Gas Chromatography (GC) and Gas Chromatography and Mass Spectrometry (GC-MS). The Antioxidant activity of the EOs were determined using Ferric Reducing Antioxidant Power (FRAP) and Total Polyphenolic Content (TPC) assays. The GC and GCMS identified 4(10)-Thujene (33.82%) as the major compound in C. medica oil, extracted by HD while that was L- terpinene-4-ol (18%) when extracted by SFME technique. The β-Linalool was the major component in C. reticulata (42.98%) and C. madurensis (37.95%) in SFME extracted EOs, however, it has been dropped down to 26.71% and 26.83% respectively during HD extraction. In C. limon, D-Limonene (43.069%) was abundant in HD- extracted EOs while it was (R)-(+)-Citronellal (39.08%) in SFME-extracted EO. SFME yielded EOs with higher amounts of more valuable oxygenated compounds with higher antioxidant capacities over HD. In comparison with HD, the SFME technique presented higher efficiency with reduced extraction times, costs and energy, yielding EOs with excellent chemical profiles and relatively high antioxidant activity.Keywords: Essential oils, Solvent free Microwave extraction, GC, GCMS, Antioxidan

Antimicrobial Properties of Ethanolic and Methanolic Extracts of Finger millet (Eleusine coracana (L.) Gaertn.) Varieties Cultivated in Sri Lanka

The emerging significance of natural antimicrobial agents creates an imperative need to identify novel plant sources with antimicrobial activities. The objective of the present study was to evaluate antibacterial, antifungal and β-lactamase enzyme inhibitory activities of ethanolic and methanolic extracts of Ravi, Rawana and Oshadha finger millet varieties. Flours of whole grains of the finger millet varieties were extracted with absolute ethanol and methanol separately. Antibacterial activities against six antibiotic-sensitive and four antibiotic-resistant pathogenic bacterial strains were evaluated using the resazurin reduction assay. Antifungal activities against six antimicrobial-sensitive pathogenic fungal strains were evaluated using the agar tube dilution method. β-Lactamase enzyme inhibitory activity was evaluated using a standard method. Both ethanolic and methanolic extracts of the three finger millet varieties showed dose-dependent inhibitory activities against the tested antibiotic-sensitive and antibiotic-resistant bacterial strains while exhibiting high inhibitions against Gram-positive antibiotic-sensitive bacterial strains when compared to Gram-negative antibiotic-sensitive bacterial strains. The findings revealed the antibacterial potential of both ethanolic and methanolic extracts of the three finger millet varieties against antibiotic-sensitive Staphylococcus aureus (ATCC® 6538™) and Bacillus subtilis (ATCC® 23857™) strains and the minimum inhibitory concentrations of the extracts against S. aureus and B. subtilis were found to be 2.1 and 1.8 mg/ml, respectively. However, none of the extracts can be considered as significantly active against the tested antibiotic-sensitive and antibiotic-resistant bacterial strains when compared to the standard drugs. In addition, none of the extracts can be considered as active against the tested fungal strains at the tested concentrations. Nevertheless, all extracts showed more activities against the tested bacterial strains when compared to the tested fungal strains. Since all extracts showed less than 40% β-lactamase inhibitory activities even at 2 mg/ml concentration, they do not qualify as promising sources of β-lactamase inhibitors at the tested concentration.KEYWORDS: Antibacterial activity, Antifungal activity, Finger millet, β-Lactamase enzyme inhibitory activit

Seasonal planning procedures to improve irrigation management performance: How Kirindi Oya experience can be transferred to NIRP schemes. Proceedings of the Workshop on Seasonal Planning Procedures to Improve Irrigation Management Performance: How Kirindi Oya experience of IIMI/ID can be transferred to NIRP schemes, held at the Irrigation Department, Colombo, Sri Lanka, on 16 May 1994

Irrigation managementIrrigation systemsMaintenanceCropsDiversificationSettlementSalinityWater use efficiencyResource managementRehabilitationPerformance

Sri Lankan Finger Millet (Elucine coracana) Variety ‘Raavana’ as Potential Probiotic Source

This study aims to isolate and identify probiotic potential lactic acid bacteria from fermented Sri Lankan finger millet variety "Raavana" and to investigate the probiotic characteristics, in vitro safety and efficacy. A bacterial isolate with typical lactic acid bacterial phenotypic and biochemical characteristics was isolated and identified. Partial sequence of the 16S rRNA gene of the Sri Lankan strain was deposited in the NCBI gene bank as Lactococcus lactis subsp. lactis FM_19LAB and the accession number MF480428 was obtained. It did not demonstrate hemolysis, DNase, gelatine hydrolysis activity as well as did not acquire complete resistance to any of the antibiotics tested hence indicating the safety. Lactococcus lactis subsp. lactis FM_19LAB had the capacity to tolerate different concentrations of acid, bile, phenol, salt, simulated gastric juices and range of temperatures. Further it exhibited anti-microbial, anticancer and anti-oxidant activities. Further, Lactococcus lactis subsp. lactis FM_19LAB assimilated cholesterol and produced lactic acid during the fermentation

Deposition of amyloid β in the walls of human leptomeningeal arteries in relation to perivascular drainage pathways in cerebral amyloid angiopathy

Deposition of amyloid beta (AB) in the walls of cerebral arteries as cerebral amyloid angiopathy (CAA) suggests an age-related failure of perivascular drainage of soluble A? from the brain. As CAA is associated with Alzheimer's disease and with intracerebral haemorrhage, the present study determines the unique sequence of changes that occur as A? accumulates in artery walls. Paraffin sections of post-mortem human occipital cortex were immunostained for collagen IV, fibronectin, nidogen 2, AB and smooth muscle actin and the immunostaining was analysed using Image J and confocal microscopy. Results showed that nidogen 2 (entactin) increases with age and decreases in CAA. Confocal microscopy revealed stages in the progression of CAA: AB initially deposits in basement membranes in the tunica media, replaces first the smooth muscle cells and then the connective tissue elements to leave artery walls completely or focally replaced by AB. The pattern of development of CAA in the human brain suggests expansion of AB from the basement membranes to progressively replace all tissue elements in the artery wall. Establishing this full picture of the development of CAA is pivotal in understanding the clinical presentation of CAA and for developing therapies to prevent accumulation of AB in artery walls. This article is part of a Special Issue entitled: Vascular Contributions to Cognitive Impairment and Dementia edited by M. Paul Murphy, Roderick A. Corriveau and Donna M. Wilcock

Isolation and identification of lactic acid bacteria with probiotic potential from fermented flour of selected banana varieties grown in Sri Lanka

This study aimed at isolating and identifying lactic acid bacteria (LAB) with probiotic potential from fermented flour of selected banana varieties grown in Sri Lanka and evaluating their probiotic and safety attributes in vitro. Ten lactic acid bacteria were isolated from six varieties of fermented banana flour, kolikuttu, seeni parakum, ambul nadee, ambum, seeni and anamalu. The isolates were screened for phenotypical and biochemical characteristics. The selected isolates were identified by 16 S rRNA sequencing as Enterococcus durans (two strains), E. gallinarum, E. hirae, E. faecium (two strains), Lactobacillus plantarum, L. curieae, Weissella cibaria and Pediococcus acidilactici and their partial sequences were deposited in NCBI. Among them, six isolates were selected based on the results of in vitro safety attributes and evaluated for their probiotic attributes. Three isolates, namely, E. durans MF405179.1, E. faecium MF574466.1 and L. curieae MF405178.1 isolated from kolikuttu, seeni parakum and ambul nadee, respectively demonstrated tolerance to acid, gastric juice, bile, salt, phenol and temperature under gastric conditions, and also showed susceptibility to tested antibiotics. Among the selected isolates, E. durans MF405179.1 demonstrated the highest hydrophobicity and auto-aggregation of 69.91 % and 76.53 %, respectively. Further, it exhibited highest adhesion to both HCT116 and HT29 cell lines demonstrating 72.5 % and 74.16 % adhesion, respectively. This is the first report of isolation and characterisation of LAB strains with probiotic potential from flour of banana varieties kolikuttu, seeni parakum, ambul nadee, ambum, seeni and anamalu grown in Sri Lanka

Dietary fibers, starch fractions and nutritional composition of finger millet varieties cultivated in Sri Lanka

Soluble, insoluble and total dietary fiber contents, rapidly and slowly digestible starch contents, arabinoxylans, β-glucans, fructans, resistant starch, amylose and total sugar contents, minerals and trace elements compositions and proximate compositions of three finger millet varieties, namely Ravi, Rawana and Oshadha, were evaluated using standard protocols. There were no significant differences (P≥ 0.05) among the rapidly digestible starch, arabinoxylans, β-glucans, fructans, amylose, total sugar, protein, crude fat and crude fiber contents of Ravi, Rawana and Oshadha varieties. Total dietary fiber contents varied between 13.01% (Ravi) and 13.79% (Oshadha). Slowly digestible starch contents ranged from 43.38% (Ravi) to 49.15% (Oshadha) and resistant starch contents ranged from 3.75% (Ravi) to 4.58% (Oshadha). Ash content of Ravi (3.22%) was significantly higher (P < 0.05) than ash contents of other two varieties. Average sodium, magnesium, potassium, calcium, iron, zinc and phosphorous contents of three finger millet varieties were 12.04, 141.78, 407.15, 345.62, 3.49, 1.89 and 331.07 mg/100 g, respectively. Findings of the present study indicated that studied finger millet varieties were good sources of dietary fibers (including resistant starch) as well as minerals and trace elements (especially potassium, calcium, phosphorous and iron) when compared to commonly consumed cereals such as rice and wheat

The Edinburgh CT and genetic diagnostic criteria for lobar intracerebral haemorrhage with cerebral amyloid angiopathy: model development and diagnostic test accuracy study

BACKGROUND: Identification of lobar spontaneous intracerebral haemorrhage associated with cerebral amyloid angiopathy (CAA) is important because it is associated with a higher risk of recurrent intracerebral haemorrhage than arteriolosclerosis-associated intracerebral haemorrhage. We aimed to develop a prediction model for the identification of CAA-associated lobar intracerebral haemorrhage using CT features and genotype.METHODS: We identified adults with first-ever intracerebral haemorrhage diagnosed by CT, who died and underwent research autopsy as part of the Lothian IntraCerebral Haemorrhage, Pathology, Imaging and Neurological Outcome (LINCHPIN) study, a prospective, population-based, inception cohort. We determined APOE genotype and radiologists rated CT imaging appearances. Radiologists were not aware of clinical, genetic, and histopathological features. A neuropathologist rated brain tissue for small vessel diseases, including CAA, and was masked to clinical, radiographic, and genetic features. We used CT and APOE genotype data in a logistic regression model, which we internally validated using bootstrapping, to predict the risk of CAA-associated lobar intracerebral haemorrhage, derive diagnostic criteria, and estimate diagnostic accuracy.FINDINGS: Among 110 adults (median age 83 years [IQR 76-87], 49 [45%] men) included in the LINCHPIN study between June 1, 2010 and Feb 10, 2016, intracerebral haemorrhage was lobar in 62 (56%) participants, deep in 41 (37%), and infratentorial in seven (6%). Of the 62 participants with lobar intracerebral haemorrhage, 36 (58%) were associated with moderate or severe CAA compared with 26 (42%) that were associated with absent or mild CAA, and were independently associated with subarachnoid haemorrhage (32 [89%] of 36 vs 11 [42%] of 26; p=0·014), intracerebral haemorrhage with finger-like projections (14 [39%] of 36 vs 0; p=0·043), and APOE ɛ4 possession (18 [50%] of 36 vs 2 [8%] of 26; p=0·0020). A prediction model for CAA-associated lobar intracerebral haemorrhage using these three variables had excellent discrimination (c statistic 0·92, 95% CI 0·86-0·98), confirmed by internal validation. For the rule-out criteria, neither subarachnoid haemorrhage nor APOE ɛ4 possession had 100% sensitivity (95% CI 88-100). For the rule-in criteria, subarachnoid haemorrhage and either APOE ɛ4 possession or finger-like projections had 96% specificity (95% CI 78-100).INTERPRETATION: The CT and APOE genotype prediction model for CAA-associated lobar intracerebral haemorrhage shows excellent discrimination in this cohort, but requires external validation. The Edinburgh rule-in and rule-out diagnostic criteria might inform prognostic and therapeutic decisions that depend on identification of CAA-associated lobar intracerebral haemorrhage.FUNDING: UK Medical Research Council, The Stroke Association, and The Wellcome Trust.</p

Secondary injury and inflammation after intracerebral haemorrhage: a systematic review and meta-analysis of molecular markers in patient brain tissue

BACKGROUND: Inflammatory responses to intracerebral haemorrhage (ICH) are potential therapeutic targets. We aimed to quantify molecular markers of inflammation in human brain tissue after ICH compared with controls using meta-analysis. METHODS: We searched OVID MEDLINE (1946–) and Embase (1974–) in June 2020 for studies that reported any measure of a molecular marker of inflammation in brain tissue from five or more adults after ICH. We assessed risk of bias using a modified Newcastle-Ottawa Scale (mNOS; mNOS score 0–9; 9 indicates low bias), extracted aggregate data, and used random effects meta-analysis to pool associations of molecules where more than two independent case–control studies reported the same outcome and Gene Ontology enrichment analysis to identify over-represented biological processes in pooled sets of differentially expressed molecules (International Prospective Register of Systematic Reviews ID: CRD42018110204). RESULTS: Of 7501 studies identified, 44 were included: 6 were case series and 38 were case–control studies (median mNOS score 4, IQR 3–5). We extracted data from 21 491 analyses of 20 951 molecules reported by 38 case–control studies. Only one molecule (interleukin-1β protein) was quantified in three case–control studies (127 ICH cases vs 41 ICH-free controls), which found increased abundance of interleukin-1β protein after ICH (corrected standardised mean difference 1.74, 95% CI 0.28 to 3.21, p=0.036, I(2)=46%). Processes associated with interleukin-1β signalling were enriched in sets of molecules that were more abundant after ICH. CONCLUSION: Interleukin-1β abundance is increased after ICH, but analyses of other inflammatory molecules after ICH lack replication. Interleukin-1β pathway modulators may optimise inflammatory responses to ICH and merit testing in clinical trials

Cognitive impairment in adults with epilepsy: The relationship between subjective and objective assessments of cognition

Aim: This study aimed to assess the relationship between objective measures of cognition and subjective perception of cognitive functioning reported by patients with epilepsy and their caregivers. Methods: One hundred patients with epilepsy attending hospital neurology outpatient clinics and their caregivers were enrolled in this study. The EpiTrack® (version 1) brief cognitive screening tool was used to measure objective impairment, the ABNAS questionnaire (A-B Neuropsychological Assessment Schedule) to assess subjective cognitive performance, and a version of the ABNAS designed to be completed by caregivers (C-ABNAS) to document caregivers' views. Patient anxiety and depression were measured using the Hospital Anxiety and Depression Scale (HADS) and considered as covariates. Patients with an uncertain diagnosis of epilepsy or likely severe comorbid mood or anxiety disorders were excluded. Results: Data from 82 patients were analyzed after exclusion of patients with uncertain diagnoses or likely severe comorbid mood or anxiety disorders. Fifty-nine (72%) had a degree of objective cognitive impairment. Fifty (84.7%) of these 59 patients had 'high' ABNAS scores concordant with the objective assessment, and 43 (72.9%) had high C-ABNAS scores matching the abnormalities detected by objective screening. Of the 23 (28%) patients without objective cognitive impairment, seven (30.4%) had concordantly low ABNAS scores, and 10 (43.4%) had concordantly low C-ABNAS scores. Patient memory impairment was more often reported by patients themselves than by caregivers (p. =. 0.011). Carers were significantly more likely to rate patients as having impaired motor coordination than patients themselves.A small part of the variance of the EpiTrack score was predicted by the C-ABNAS.Objective cognitive performance did not predict ABNAS or C-ABNAS scores. Conclusions: Self-report or caregiver report questionnaires identify patients with epilepsy and objective cognitive impairment more accurately than patients with intact cognition. Those without objective evidence of cognitive impairment may, nevertheless, perceive themselves as having memory dysfunction; it is these patients, therefore, who most require both subjective and objective assessments of cognition, including carers' assessments, in order to establish the nature of their symptoms. None of these assessment measures can be used as a reliable proxy for another, each contributes individually to a comprehensive assessment of cognition, and all must be used in conjunction with measures of mood and anxiety