Research on Pathogenic Fungi and Mycotoxins in China (Volume II).

The presence of pathogenic fungi and contamination of mycotoxins in food and feed pose significant threats and challenging issues to food in the world [...].

Characterization and toxicological potential of Alternaria alternata associated with post-harvest fruit rot of Prunus avium in China.

Post-harvest fruit rot caused by Alternaria species is one of the most important threats to the fruit industry. Post-harvest rot on sweet cherry (Prunus avium) fruit was observed in the fruit markets of the Haidian district of Beijing, China. The fungal isolates obtained from the infected sweet cherry fruits matched the descriptions of Alternaria alternata based on the morphology and multi-gene (ITS, endo-PG, and Alta1) sequence analysis. Pathogenicity tests indicated that ACT-3 was the most virulent isolate, exhibiting typical post-harvest fruit rot symptoms. Physiological studies revealed that the optimal conditions for the growth of ACT-3 were temperature of 28°C, water activity of 0.999, and pH of 8 with 87, 85, and 86 mm radial growth of ACT-3 on a potato dextrose agar (PDA) medium, respectively, at 12 days post-inoculation (dpi). Moreover, the fungus showed the highest growth on a Martin agar medium (MAM) modified (85 mm) and a PDA medium (84 mm) at 12 dpi. The proliferation of the fungus was visualized inside the fruit tissues by confocal and scanning electron microscope (SEM), revealing the invasion and destruction of fruit tissues. Alternaria mycotoxins, tenuazonic acid (TeA), and alternariol (AOH) were detected in five representative isolates by HPLC analysis. The highest concentrations of TeA (313 µg/mL) and AOH (8.9 µg/mL) were observed in ACT-6 and ACT-3 isolates, respectively. This study is the first to present a detailed report on the characteristics and proliferation of A. alternata associated with sweet cherry fruit rot and the detection of toxic metabolites.

Discordance in exclusive breastfeeding between maternal recall and deuterium dose-to-mother technique during the first 6 months of infants: A multi-country study in Asia.

PURPOSE: This study aimed to assess the agreement in EBF between maternal recall and the dose-to-mother (DTM) technique. METHODS: Indonesia, Malaysia, Mongolia, Pakistan, Sri Lanka, Thailand, and Vietnam participated in the study. A total of 207 and 118 mother-infant pairs were assessed at 3 and 6 months of child's age. Using a standardized questionnaire, mothers were asked to recall child feeding during the previous 24 h, at 3 and 6 months. Those recalled to be EBF proceeded to be assessed using DTM technique. Non-milk oral intake (NMOI) cutoff of 86.6 g/d was used to classify EBF. RESULTS: According to DTM, 66% of infants were EBF at 3 months, while only 22% were EBF at 6 months. At 3 months, the overall % agreement between maternal recall and DTM method was 68%, kappa 0.06 (95% CI: 0.07-0.20), and at 6 months, the % agreement was only 21%, kappa -0.031 (95% CI -0.168 to 0.107). Human milk intakes were similar at 3 months and 6 months when expressed as g/d, but decreased when expressed as g/kg/d, with a large variation within and between countries; Pakistan being the lowest. CONCLUSION: This study showed there were declining levels of EBF from 3 to 6 months in the participating countries from Asia and the agreement between maternal recall and DTM technique to classify EBF was low. To ensure that the DTM technique can be more widely used in evaluating breastfeeding promotion programs, consensus on the appropriate NMOI cutoff and simplification of the DTM protocol is necessary.

Phytocompound screening, antioxidant activity and molecular docking studies of pomegranate seed: a preventive approach for SARS-CoV-2 pathogenesis.

A global hazard to public health has been generated by the coronavirus infection 2019 (COVID-19), which is spreading quickly. Pomegranate is a strong source of antioxidants and has demonstrated a number of pharmacological characteristics. This work was aimed to analyze the phytochemicals present in ethanolic pomegranate seed extract (PSE) and their in vitro antioxidant potential and further in-silico evaluation for antiviral potential against crystal structure of two nucleocapsid proteins i.e., N-terminal RNA binding domain (NRBD) and C-terminal Domain (CTD) of SARS-CoV-2. The bioactive components from ethanolic extract of PSE were assessed by gas chromatography-mass spectroscopy (GC-MS). Free radical scavenging activity of PSE was determined using DPPH dye. Molecular docking was executed through the Glide module of Maestro software. Lipinski's 5 rule was applied for drug-likeness characteristics using cheminformatics Molinspiration software while OSIRIS Data Warrior V5.5.0 was used to predict possible toxicological characteristics of components. Thirty-two phytocomponents was detected in PSE by GC-MS technique. Free radical scavenging assay revealed the high antioxidant capacity of PSE. Docking analysis showed that twenty phytocomponents from PSE exhibited good binding affinity (Docking score ≥ - 1.0 kcal/mol) towards NRBD and CTD nucleocapsid protein. This result increases the possibility that the top 20 hits could prevent the spread of SARS-CoV-2 by concentrating on both nucleocapsid proteins. Moreover, molecular dynamics (MD) simulation using GROMACS was used to check their binding efficacy and internal dynamics of top complexes with the lowest docking scores. The metrics root mean square deviation (RMSD), root mean square fluctuation (RMSF), intermolecular hydrogen bonding (H-bonds) and radius of gyration (Rg) revealed that the lead phytochemicals form an energetically stable complex with the target protein. Majority of the phytoconstituents exhibited drug-likeness with non-tumorigenic properties. Thus, the PSE phytoconstituents could be useful source of drug or nutraceutical development in SARS-CoV-2 pathogenesis.

TeM-DTBA: time-efficient drug target binding affinity prediction using multiple modalities with Lasso feature selection.

Drug discovery, especially virtual screening and drug repositioning, can be accelerated through deeper understanding and prediction of Drug Target Interactions (DTIs). The advancement of deep learning as well as the time and financial costs associated with conventional wet-lab experiments have made computational methods for DTI prediction more popular. However, the majority of these computational methods handle the DTI problem as a binary classification task, ignoring the quantitative binding affinity that determines the drug efficacy to their target proteins. Moreover, computational space as well as execution time of the model is often ignored over accuracy. To address these challenges, we introduce a novel method, called Time-efficient Multimodal Drug Target Binding Affinity (TeM-DTBA), which predicts the binding affinity between drugs and targets by fusing different modalities based on compound structures and target sequences. We employ the Lasso feature selection method, which lowers the dimensionality of feature vectors and speeds up the proposed model training time by more than 50%. The results from two benchmark datasets demonstrate that our method outperforms state-of-the-art methods in terms of performance. The mean squared errors of 18.8% and 23.19%, achieved on the KIBA and Davis datasets, respectively, suggest that our method is more accurate in predicting drug-target binding affinity.

Body composition reference charts for infants from birth to 24 months: Multicenter Infant Body Composition Reference Study.

BACKGROUND: Body composition assessment in the first 2 y of life provides important insights into child nutrition and health. The application and interpretation of body composition data in infants and young children have been challenged by a lack of global reference data. OBJECTIVES: We aimed to develop body composition reference charts of infants aged 0-6 mo based on air displacement plethysmography (ADP) and those aged 3-24 mo based on total body water (TBW) by deuterium dilution (DD). METHODS: Body composition was assessed by ADP in infants aged 0-6 mo from Australia, India, and South Africa. TBW using DD was assessed for infants aged 3-24 mo from Brazil, Pakistan, South Africa, and Sri Lanka. Reference charts and centiles were constructed for body composition using the lambda-mu-sigma method. RESULTS: Sex-specific reference charts were produced for FM index (FMI), FFM index (FFMI), and percent FM (%FM) for infants aged 0-6 mo (n = 470 infants; 1899 observations) and 3-24 mo (n = 1026 infants; 3690 observations). When compared with other available references, there were observable differences but similar patterns in the trajectories of FMI, FFMI, and %FM. CONCLUSIONS: These reference charts will strengthen the interpretation and understanding of body composition in infants across the first 24 mo of life.

Biocontrol potential of lipopeptides produced by the novel Bacillus subtilis strain Y17B against postharvest Alternaria fruit rot of cherry.

The use of synthetic fungicides against postharvest Alternaria rot adversely affects human health and the environment. In this study, as a safe alternative to fungicides, Bacillus subtilis strain Y17B isolated from soil exhibited significant antifungal activity against Alternaria alternata. Y17B was identified as B. subtilis based on phenotypic identification and 16S rRNA sequence analysis. To reveal the antimicrobial activity of this strain, a PCR-based study detected the presence of antifungal lipopeptide (LP) biosynthetic genes from genomic DNA. UPLC Q TOF mass spectrometry analysis detected the LPs surfactin (m/z 994.64, 1022.68, and 1026.62), iturin (m/z 1043.56), and fengycin (m/z 1491.85) in the extracted LP crude of B. subtilis Y17B. In vitro antagonistic study demonstrated the efficiency of LPs in inhibiting A. alternata growth. Microscopy (SEM and TEM) studies showed the alteration of the morphology of A. alternata in the interaction with LPs. In vivo test results revealed the efficiency of LPs in reducing the growth of the A. alternata pathogen. The overall results highlight the biocontrol potential of LPs produced by B. subtilis Y17B as an effective biological control agent against A. alternata fruit rot of cherry.

A machine learning strategy with clustering under sampling of majority instances for predicting drug target interactions.

Drug Target Interactions (DTIs) are crucial in drug discovery as it reduces the range of candidate searches, speeding up the drug screening process. Considering in vitro and in vivo experimentations are time and cost-expensive, there has been a surge in computational techniques, especially ML methods for DTIs prediction. Therefore, this study aims to present a methodology that uses molecular structures and amino acid sequences for generating PSSM and PubChem fingerprints for drugs and targets respectively. The proposed work uses a novel technique NearestCUS for handling the class imbalance problem of the benchmark datasets. We use Isomap Embedding to extract features from PSSMs. Feature selection is performed using ANOVA. CatBoost is used for predicting the interaction between drugs and targets for the first time. To quantify the efficacy of NearestCUS, we compared it with other sampling techniques. We found that the proposed methodology performed better than state-of-the-art approaches.

Bacillus thuringiensis CHGP12 uses a multifaceted approach for the suppression of Fusarium oxysporum f. sp. ciceris and to enhance the biomass of chickpea plants.

BACKGROUND: Bacillus species synthesize antifungal lipopeptides (LPs) making them a sustainable and eco-friendly management option to combat Fusarium wilt of chickpea. RESULTS: In this study, 18 endophytic Bacillus strains were assessed for their antifungal activity against Fusarium oxysporum f. sp. ciceris (FOC) associated with Fusarium wilt of chickpea. Among them, 13 strains produced significant inhibition zones in a direct antifungal assay while five strains failed to produce the inhibition of FOC. Bacillus thuringiensis CHGP12 exhibited the highest inhibition 3.45 cm of FOC. The LPs extracted from CHGP12 showed significant inhibition of the pathogen. Liquid chromatography-mass spectrometry (LC-MS) analysis confirmed that CHGP12 possessed the ability to produce fengycin, surfactin, iturin, bacillaene, bacillibactin, plantazolicin, and bacilysin. In an in vitro qualitative assay CHGP12 exhibited the ability to produce lipase, amylase, cellulase, protease, siderophores, and indole 3-acetic acid (IAA). IAA and gibberellic acid (GA) were quantified using ultra-performance liquid chromatography (UPLC) with 370 and 770 ng mL-1 concentrations of IAA and GA respectively. Furthermore, the disease severity showed a 40% decrease over control in CHGP12 treated plants compared to the control in a glasshouse experiment. Moreover, CHGP12 also exhibited a significant increase in total biomass of the plants namely, root and shoot growth parameters, stomatal conductance, and photosynthesis rate. CONCLUSION: In conclusion, our findings suggest that B. thuringiensis CHGP12 is a promising strain with high antagonistic and growth-promoting potential against Fusarium wilt of chickpea. © 2022 Society of Chemical Industry.

Eco-friendly acid dyeing of silk and wool fabrics using Acid Violet 49 dye.

The effluent load from textile industries has forced traders and industrialists to use sustainable tools that not only save energy, money, and labor but also make the process cleaner. The purpose of this study is to improve the dyeing of proteinous fabrics using Acid Violet 49 dye under microwave radiation. Aqueous and acidic dye solutions were prepared and treated with MW radiations for up to 10 min. MW treated and untreated dye solutions were used to color treated and untreated fabrics to observe color yield. It has been found that dyeing of irradiated silk at 65 °C for 35 min, using 55 mL of irradiated dye solution containing 1 g/100 mL salt, has given excellent results. Whereas good color characteristics are obtained if irradiated wool fabric is dyed at 85 °C for 55 min, using 55 mL irradiated dye solution using 1 g/100 mL salt. Physicochemical analysis reveals that MW rays have physically modified the fabric without altering its chemistry. ISO standard methods employed for colorfastness show that under optimal conditions, the color developed is fast; statistical analysis shows that the dyeing process has given significant results. It is concluded that MW rays have excellent potential to improve acidic dyeing of proteinous fabric under mild conditions which show that the utilization of MW rays is a cost-, time-, and energy-effective process.

Prevalence of irritable bowel syndrome and its associated risk factors among university students of Bangladesh.

Background: Irritable bowel syndrome (IBS) is a very common gastrointestinal disorder worldwide, but research regarding this disease is rare in Bangladesh. This study aimed to assess the prevalence of IBS and its associated risk factors among university students in Bangladesh. Methods: This is a cross-sectional study. A total of 300 randomly selected participants were included in this study. By using a structured questionnaire and anthropometric methods, we collected all the required data for our study. The diagnosis of IBS was based on Rome III criteria. Results: The overall prevalence of IBS was 39.3%, but the majority (77.3%) had no basic awareness of IBS. In our study, anxiety and depression (χ 2 = 6.817; odds ratio [OR] = 1.910; 95% confidence interval [CI] = 1.172, 3.113; P = 0.011) had a significant relationship with IBS and IBS had a significant (P < 0.001) relationship with food intolerance (χ 2 = 8.737; OR = 2.130; 95% CI = 1.284, 3.531), chest pain (χ 2 = 7.482; OR = 2.035; 95% CI = 1.218, 3.401), and insomnia (χ 2 = 19.320; OR = 2.907; 95% CI = 1.794, 4.709). In our dietary data, the intake patterns of vegetables (P = 0.000), fast food (P = 0.000), and tea-coffee (P = 0.003) showed a strong significant association with IBS. On the other hand, monthly household income (P = 0.154) and body mass index (BMI) (P = 0.138) showed no significant association with IBS. Among our study subjects, IBS-constipation (54.2%) was more common than IBS-diarrhea (27.1%) and IBS-mixed (18.6%). Moreover, among the 118 IBS respondents, 67.8% had a headache with increased flatulence (95.8%) as the most common IBS-related complication. Conclusion: IBS is common in university students of Bangladesh and is associated with anxiety, depression, and particular dietary patterns.

Sustainable eco-friendly extraction of yellow natural dye from haar singhar (Nyctanthes arbor-tritis) for bio coloration of cotton fabric.

The revival of natural dyes in different walks of life is due to stringent environmental standards imposed by many associations. For current studies, flowers of haar singhar (Nyctanthes arbor-tritis) has been chosen for bio-dyeing of cotton fabric using microwave irradiation techniques. For this purpose, liquid extracts and fabrics were exposed to microwave for 5 min at various conditions. These treated and un-treated dye extracts obtained in respective media were employed to dye the radiated and non-radiated cotton fabrics. The characterization of extract and untreated and undyed irradiated fabrics samples was done through FTIR. Different dyeing variables were optimized under CCD response surface methodology as a statistical tool. With the introduction of new shades and improvement of colorfastness properties, different concentrations of sustainable chemical and bio-mordant were employed. All dyed cotton fabrics were exposed to CIE-color space system for estimation of color coordinates and color strength by using spectra flash SF600 and to rate colorfastness properties using ISO standard methods for light, dry and wet rubbing, washing fastness. Cotton fabric was dyed with haar singhar flower extract of 7 pH from 4 g of powder, using 1.5 g/100 mL salt solution at 60 °C for 30 min. To get acceptable shades, aqueous extract after microwave treatment for 4 min provided excellent color strength. Pistachio (K/S = 3.6342) is a bio-mordant with great results, and aluminum (K/S = 4.8205) is a chemical mordant with outstanding results. It is found that green methods for isolation of colorant and green mordants for getting new shades should be employed.

First Report of Bacterial Leaf Spot of Ficus benghalensis Caused by Pseudomonas cichorii in Pakistan.

Ficus benghalensis L. belongs to the family Moraceae, native to Asia and commonly known as Banyan. It has been identified as an important medicinal tree due to its antioxidant, anti-diabetic, and anti-inflammatory properties (Singh et al., 2009; Thite et al., 2014). In March 2021, leaf spots were observed on Banyan trees in the Kharian forest zone, District Gujrat, Punjab Province, Pakistan. Initial symptoms on leaves were irregular, water-soaked, and light brown lesions. The lesions turned dark brown at the centre, and the margins gradually turned yellow. The average size of lesions was 12 to 20 × 8 to 13 mm. The lesions coalesced and produced necrotic areas on the leaf (Figure 1). Samples (n=34) were collected based on symptoms and infected leaf segments were excised into small pieces (10-20 mm), surface disinfected with 1% NaClO for 10 seconds and rinsed three times with sterilized distilled water (SDW). Ten pieces/sample were mashed and soaked in 1.5 ml SDW to obtain a suspension. Later, 10 µL of the suspension was streaked on Nutrient agar (NA) and King's B medium (KBM) and incubated for 72 h at 30°C. After 72 h bacterial colonies appeared on NA and KBM medium. Each colony was re-streaked for three times to obtain the purified colonies. Morphological and biochemical characteristics of isolated bacterial cultures were performed by following the method of Schaad et al. (2001). Bacterial colonies appeared pale yellow to creamy, smooth, and circular with undulated margins on both NA and KBM medium. The colonies produced a fluorescent blue colour on KBM under the UV light. Isolated bacterial cultures were positive for oxidase, negative for levan production and arginine dihydrolase. Based on these characteristics, the pathogen was identified as Pseudomonas species. For molecular identification, 16S rRNA and rpoB genes were amplified and sequenced using the following primers: 27F/1492R (Lane, 1991) and LAPS/LAPS27 (Ait Tayeb et al. 2005), respectively. All the isolates were identified as P. cichorii after BLASTn analysis. The sequences of isolate BLS-01 obtained in this study were deposited in GenBank with accession No. OK397593 for 16S rRNA and OK423684 for rpoB exhibiting 100 % similarity with P. cichorii Accession No. MK356431 for 16S rRNA and JQ267563 for rpoB. A pathogenicity test was performed on healthy Banyan seedlings to fulfil Koch's postulates. Leaves from seedling plants were inoculated with 3 mL of BLS-01 suspension (108 CFU/ml) by spraying the inoculum on leaves using a sterilized spray bottle. The leaves sprayed with sterilized distilled water served as control (Figure 2). The experiment was performed three times following the same protocol as described above. Symptoms that appeared on inoculated leaves after 7-10 days were similar to the symptoms observed on original infected Banyan tree leaves in the forest zone. Control leaves remained asymptomatic during the whole experiment. The pathogen from the artificial infected leaves was re-isolated and identified as P. cichorii based on morphological, biochemical, and molecular characteristics. To our knowledge, this is the first report of leaf spot of F. benghalensis caused by P. cichorii in Pakistan.

First record of Leaf Spot of Ficus religiosa Caused by Alternaria alternata in Pakistan.

Ficus religiosa (L.) belongs to the family Moraceae, native to India and commonly known as 'Peepal'. It has high medicinal value due to its antibacterial, antiviral and antioxidant properties (Singh et al., 2015; Kalpana et al., 2009). In August 2021, leaf spots were observed on F. religiosa trees in Pabbi forest park Kharian (32°50'01.4"N 73°50'17.7"E), District Gujrat, Pakistan. The disease incidence was recorded approximately 30%. The leaf spots were irregular in shape, brown in colour, 3-9 mm in size and encircled by yellowish halo. In severe condition, the spots coalesced and produced necrotic areas on leaf surface (Figure 1). The samples (n=21) were collected based on symptoms and infected leaf segments were excised into small pieces, surface disinfected with 1% NaClO for 20s and rinsed 3 times with sterilized distilled water. The pieces were plated on Potato Dextrose Agar (PDA) medium and incubated at 28°C for 7 days. All the pure cultures were obtained through single spore method on PDA and preserved in 30% glycerol at -80°C. The colonies were olive green to dark brown with white margin and later turned dark olive or black with enormous sporulation. Conidia (n=24) were obclavate, ovoid, brown in colour and measuring 10.2 to 34.1 µm long x 5.9 to 12.3 µm wide with 1 to 6 transverse and 1 to 3 longitudinal septa (Figure 2). Based on these characteristics, the pathogen was identified as Alternaria alternata (Gilardi, G., et al. 2019). For molecular identification, the Internal Transcribed Spacers (ITS) region, endopolygalacturonase (endoPG) gene and major allergen (Alt a1) gene were amplified using ITS1/4 (White et al. 1990), PG3/PG2b (Andrew et al. 2009) and Alt-4for/Alt-4rev (Lawrence et al. 2013) primers respectively. Based on molecular characteristics, all isolates were identified as A. alternata. The sequences of the representative isolate FLB-1 were submitted in the GenBank with the accession numbers OL514181 for ITS, OK315658 for endoPG and OK315659 for Alt al showing 100% similarity with ITS accession KP124298, and endoPG accession AY205020 and 99.7% with Alt al accession KP123847 sequences of CBS106.24 A. alternata after BLASTn queries. The Phylogenetic reconstruction based on maximum likelihood, using Mega X (Kumar et al. 2018) and FLB-1 grouped with A. alternata (Figure 3). Pathogenicity test was performed on nine months old healthy F. religiosa (L.) seedlings (n=12) to fulfil the Koch's postulates. The leaves were pinpricked and sprayed with FLB-1 conidial suspension (107 spores/ml) by using atomizer (Bajwa et al., 2010). The leaves of F. religiosa (L.) seedlings (n=12) sprayed with sterilized distilled water served as control. All the seedlings were incubated at 25 ± 3°C in the glasshouse. The experiment was performed three times under the same conditions. The typical symptoms appeared on inoculated leaves after 7-14 days that were similar to the symptoms observed on original infected F. religiosa (L.) trees. In the control treatment leaves remained asymptomatic (Figure 4). The pathogen from the artificial infected leaves was re-isolated and identified as A. alternata based on morphological and molecular characteristics. To our knowledge, this is the first report of leaf spot of F. religiosa (L.) caused by A. alternata in Pakistan. The leaves of F. religiosa (L.) are commonly used in Asia for different purposes and this leaf spot disease may represent a significant threat to F. religiosa (L.) tree health.

First report of leaf blight disease of Livistona chinensis caused by Alternaria alternata in Pakistan.

Livistona chinensis (Jacq.) is also known as fan palm and is commonly grown in the subtropical region of the world. This plant is widely cultivated in Asia for ornamental purpose and also used in Chinese medicines (Li et al. 2019). In May 2021, severe leaf blight was observed on L. chinensis leaves in ornamental plant nurseries, located at Pattoki (30°59'41.5"N 73°48'43.8"E) District Kasur, Punjab province, Pakistan. The disease incidence was up to 50% and the initial symptoms appeared as chlorotic brown spots on the upper portion of leaves. Later, the spots expanded and changed into elliptical lesions on the leaves. The lesions with dark brown margins coalesced to cause extensive tissue necrosis of leaves and exhibited blight (Figure 1). Two to three leaves were taken from each infected plant. Infected leaves of each sample of L. chinensis were excised into small pieces (3-4 mm) with the help of sterilized scissor and surface disinfected with 1% NaClO for 20s and rinsed 3 times with sterilized distilled water. To isolate the potential causal organism, these pieces were plated on Potato Dextrose Agar (PDA) medium and incubated at 28 °C with 70 % relative humidity for 7 days. Purified cultures were obtained through single spore culture on PDA. All obtained isolates were preserved in 30% glycerol at -80°C. The fungal colony colour was olive to dark greenish and dark brown to black on the reverse side. The conidia (n=36 per isolate) were greenish to brown in colour, ellipsoid to obclavate, ovoid, irregular and measured an average range from 10.9 to 30.7 µm long x 6.3 to 12.5 µm wide with 2 to 5 transverse and 0 to 3 longitudinal septa (Figure 2). The genomic DNA was extracted from all isolates (n=40) and multi-locus sequence analysis approach was used for molecular identification. The Internal Transcribed Spacers (ITS) region, Alt a1 major allergen (ALT) gene, glyceraldehyde-3-phosphate dehydrogenase (GPD), actin (ACT) gene and histone 3 genes were amplified using ITS1/4 (White et al. 1990), Alt-4for/Alt-4rev (Lawrence et al. 2013), GPD1/GPD2 (Guerber et al. 2003), ACT512F/ACT783R (Carbone and Kohn, 1999) and H3-1a/H3-1b (Luan et al. 2007). Based on morphological and molecular characteristics, all isolates were identified as Alternaria alternata. The sequences of the representative isolate APLB-3 were submitted in the GenBank with the accession numbers (ITS: MZ663802), (ALT: MZ666883), (ACT: MZ666885), (GPD: MZ666884), and (Histone3: MZ666886) showing 100% similarity with ITS accession MK968038, ALT accession MN702781, ACT accession MT318253, GPD accession MT524743 and histone 3 accession MH824369. For pathogenicity test, potted L. chinensis plants (n=9) leaves were pin-pricked with sterilized needle (Bajwa et al. 2010) and inoculated with spore suspensions (107 spore/ml) of APLB-3 (1ml/leaf) to confirm Koch's postulates. After 14 days, the inoculated leaves showed chlorotic brown spots and leaf blight symptoms similar to those observed on infected plants in nurseries. The plants grown as the control group (n=9) were sprayed with sterilized distilled water and had no symptoms (Figure 3). The experiment was performed three times. The fungal pathogen was re-isolated from the artificial inoculated leaf tissues and identified as A. alternata based on morphological and molecular characterization. To our knowledge, this is the first record of A. alternata causing leaf blight disease of L. chinensis in Pakistan. This disease may potentially decrease the value of ornamental plants in Pakistan under favourable conditions and proper management strategies should be applied.

Assessment of dietary habits, nutritional status and common health complications of older people living in rural areas of Bangladesh.

BACKGROUND: Old age is one of the vulnerable and prone stages in terms of health status. So this study aimed to assess the nutritional status and common health complications of older people. METHODS: Simplified Nutritional Appetite Questionnaire (SNAQ), Anthropometric measurements, Diet History Method, and Mini Nutritional Assessment (MNA) tools were used to measure the nutritional status. Data were analyzed by using Statistical Package for Social Science (SPSS) version 16. RESULTS: Out of the total 320 elderly participants the mean ± SD value for the age of male and female was 67.25 ± 6.5 and 67.32 ± 7.7 years respectively. According to BMI classification, it was noticed that with advancing age the percentage of underweight was also increased such as for 60-75 years old age group the underweight percentage was 30.0% where for 76 to 85 and >85 years old age group the underweight percentage was 45.0% and 60.0% respectively. According to the MNA score, 97 elderly respondents were malnourished and a total of 172 respondents had SNAQ scores below 14. This study found a statistically significant (P < 0.05) correlations among various health complications with nutritional status according to MNA score. In addition 56.6% (OR = 1.24, 95% CI = .799-1.939), 63.8% (OR = 1.18, 95% CI = .745-1.857) and 64.7% (OR = 1.14, 95% CI = .720-1.804) respondents had diabetes mellitus, hypertension and cardiovascular disease respectively. The risk of musculoskeletal pain (OR = 1.073, 95% CI = .684-1.681), bedsore (OR = 1.884, 95% CI = .903-3.934) and decreased sense of thirst (OR = 1.278, 95% CI = .821-1.991) were higher among females than males. A little number of the elderly used to take milk, meat, and fish daily. CONCLUSION: During this cross-sectional study, significant correlations among nutritional changes with health complications were determined. To prevent malnutrition among the elderly a proper health policy as well as periodical nutritional screening should be conducted.

Distributed renewable energy systems for resilient and sustainable development of remote and vulnerable communities.

More than 940 million people (13% of the world population) do not have any access to electricity. Most of these people live in rural and remote areas, where the lack of electricity access is significantly damaging the quality of life, economic development and the environment. Distributed energy systems (DESs) (based on clean energy technologies) for energy access offer a potentially important strategy for pursuing environment-friendly sustainable development and poverty alleviation; especially in rural and remote communities. DESs are also helpful in reducing deforestation and greenhouse gas (GHG) emissions as the fossil fuel-based energy production is among the largest contributors to GHG emissions. This paper presents the importance of energy access through DESs for resilient and sustainable development using two projects case studies from Pakistan. The first project case study is based on the Afghan refugee villages, where, currently, there is no electricity access. A baseline survey was conducted to assess the socio-economic conditions and energy demand of the refugees. This assessment is then used to devise clean energy solutions as per the local context. This project aims to improve the quality of life of the refugees by providing energy access. In the second case study, electricity access was provided to the local communities a few years ago. Analyses of primary data collected in this case study show that DES integrated with socio-economic and cultural systems can bring a significantly positive impact on the local communities, advancing all the sustainability development goals. This work concludes that DES can be significantly generative, if effectively integrated into socio-economic processes. This article is part of the theme issue 'Developing resilient energy systems'.

Data mining and machine learning in cancer survival research: An overview and future recommendations.

Data mining and machine learning techniques are transforming the decision-making process in the medical world. From using nomograms and expert advice, scientists are now moving towards machine learning and deep learning techniques to make informed decisions for patients. The change in this aspect is mainly attributed to large amounts of digital data stored in hospitals. This study is focused on the transformation of cancer survival research in the past few years. A road map based on seven different aspects has been provided in this study utilizing various machine learning techniques, presenting a review of 62 articles published in the past 15 years. It was found that researchers are now moving to more clinical data even with less number of instances. Though most of the studies used traditional machine learning techniques for predicting cancer survival, researchers are now moving towards deep learning and hybrid approaches to gain some insights into survival prediction. Finally, this study presents ten new open research issues and possible future research plans to focus on for better results in cancer survival research. It is hoped that this review will be viewed by both apprentice and expert researchers as a valuable resource to understand the currently used practices and possible future recommendations to work.

Eco-friendly approach towards isolation of colorant from Esfand for bio-mordanted silk dyeing.

Sustainability in all applied fields particularly in textiles is to protect our globe, environment, and community, where green dyed products are playing their role. For the current study, Esfand (Peganum harmala) has been explored using a green isolation tool, i.e., ultrasonic (U.S.) rays, and applied onto fabric. Different dyeing parameters have been explored statistically through response surface methodology by employing temperature (50-80°C), time (25-65 min), extract volume (15-55 mL), salt (1-5 g/100 mL), and dye bath pH (4-7) through series of experiments. For developing new shades, green mordants such as elaichi, neem, turmeric, and zeera have been utilized. It has been found that exposure of 35 mL extract of 7 pH containing 3 g/100 mL of salt as exhausting agent to U.S. rays for 30 min for the dyeing of silk at 70°C for 45 min has given maximum color strength with reddish-yellow shades. Color characteristics obtained in the CIE Lab system reveal that 5% of turmeric as meta bio-mordant has given good quality reddish-yellow shades. It is found that U.S. rays have not only good potential to isolate colorant followed by dyeing of silk under reduced condition but also the application of bio-mordants have made the process more greener, sustainable, and cleaner.