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The study is to evaluate the impact of COVID-19 in the Pediatric Oncology Units (POUs) of Pakistan. Data from 1 April 2019 to 30 June 2019 and 1 April 2020 to 30 June 2020 for the first and second cohort, respectively, in order to compare the registration, abandonment rate, and delay in treatment. Six hundred and thirty-four were registered cases, 379 and 255 in the first and second cohort, respectively, which was significantly different0.005. Seventy-seven were abandoned, 45 and 32 in the first and second cohort, respectively. Fifty-nine COVID-19 positive cases, 24, 4, 27, and 4 were admitted, referred, home isolated, and leave against medical advice (LAMA), respectively. Delayed treatment and reduction in new cases were observed.

Objective: To determine the impact of endemicity of ascariasis in the manifestation of acute appendicitis. Study Design: Prospective observational study. Place and Duration of Study: General Surgery Department, Combined Military Hospital Skardu, from May 2012 to Apr 2015. Methodology: Patients presenting with symptoms of acute appendicitis were included in the study. Patients were diagnosed after taking detailed history, clinical examination and blood chemistry. All the patients underwent open appendectomy under general anesthesia. Per-operatively presence of worms was assessed in terminal ileum and after that in the lumen of the appendix initially by palpation and then by opening its lumen after it was removed. Histopathologic diagnosis could not be confirmed due to its non-availability as the surgeries were performed in a remote and resource constraint area of the country. Results: A total of 224 patients were included in the study. Out of these, 143 (63.8%) were male and 81 (36.1%) female patients. The mean age of patients was 22 ± 6 years. Per-operatively, 21 (9.3%) patients had worms in both ileum and appendicular lumen while 22 (9.8%) patients had worms only in the terminal ileum and there were no worms seen in either ileum or appendix in 181 (80.8%) patients. Conclusion: There is a relation of parasitic infestation especially Ascaris lumbricoides in the manifestation of acute appendicitis in endemic areas.

Present research exertion provides the information about the stratigraphy, geological structure, geochemistry and petrographic characteristics of the exposed sedimentary rock units and igneous intrusions around Zhob and Qila Saifullah districts of Balochistan. Sedimentary deposition in this area is attribute to both Pashin basin and Suleiman basin. The litho-stratigraphic sections of Suleiman basin are Spingwar formation and Loralai formation (Allozai Group) of Jurassic age and Goru formation, Parh limestone and Pab sandstone of Cretaceous age, whereas Pashin basin comprises of Nisai formation of Eocene age and Multana formation of 0ligocene - Miocene age. The structural features of the area are comprising of complex, doubly plunging, anticlines and synclines which have been truncated by reverse faults at different places.Geochemical analysis (X-Ray diffraction and X-Ray fluorescence) displays that Igneous Intrusions are comprising of Clay, Mica, Plagioclase and Spinal mineral with abundance of Al203 (11-14% wt.), Si02 (27-34 % wt.), Ca0 (6-17% wt.) and Fe203 (19-32% wt.), whereas sedimentary rocks are rich in Quartz, Calcite and Albite with abundance of Al203 (0-17% wt.), Si02 (1.35-78% wt.), Ca0 (0.92-55% wt.) and Fe203 (0.07-13% wt.). The petrographic studies of the carbonates indicate mudstone to wackstone, carbonate fabric with skeletal grains of Alveolina, Miliolids, Globotrancana, Algae, Stylolites and foraminifera, whereas the sandstone samples are rich in sub rounded to rounded quartz grain, feldspar and mica. Petrographic analysis of igneous intrusion shows muscovite, pyroxene needles, chlorite and opaque minerals. Economically the research area is appropriate for cement factory, glass industry and building stones.

Bulbine natalensis, an emerging medicinal herb on the global market with androgenic properties, is often formulated in dietary supplements that promote perceived sexual enhancement. However, to date, comprehensive safety studies of B. natalensis are lacking, particularly those related to its herb-drug interaction potential. The purpose of this study was to assess the inductive and inhibitory effects of extracts and pure compounds of B. natalensis on human cytochrome P-450 isozymes in vitro. Our findings demonstrated that both water and methanolic extracts of B. natalensis as well as knipholone, bulbine-knipholone, and 6′-O-methylknipholone dose-dependently increased mRNA expression encoded by CYP2B6, CYP1A2, and ABCB1 genes. Functional analyses showed that water (60 to 2.20 µg/mL) and methanolic (30 to 3.75 µg/mL) extracts and knipholones (10 to 0.33 µM) increased CYP2B6 and CYP1A2 activity in a dose-dependent manner. Additionally, water extract (60 µg/mL), methanolic extract (30 µg/mL), and knipholone (10 µM) caused activation of the aryl hydrocarbon receptor up to 11.1 ± 0.7, 8.9 ± 0.6, and 7.1 ± 2.0-fold, respectively. Furthermore, inhibition studies revealed that methanolic extract attenuated the activity of metabolically active CYP1A2 (IC50, 22.6 ± 0.4 µg/mL) and CYP2B6 (IC50, 34.2 ± 6.6 µg/mL) proteins, whereas water extracts had no inhibitory effect on either isoform. These findings suggest that chronic consumption of B. natalensis may affect normal homeostasis of select CYPs with subsequent risks for HDIs when concomitantly ingested with conventional medications that are substrates of CYP2B6 and CYP1A2. However, more in-depth translational studies are required to validate our current findings and their clinical relevance.

Two monobenzoylcyclopropane (hypoxhemerol A (1) and hypoxhemeroloside G (2)) and three dibenzoylcyclopropane (hypoxhemerol B (3), hypoxhemeroloside H (4), and hypoxhemeroloside I (5)) derivatives were isolated from the hydro-alcoholic extract of Hypoxis hemerocallidea corms. This is the first instance where benzoylcyclopropane analogs were isolated from any natural source. Structure elucidation was mainly based on 1D- and 2D-NMR and HRESIMS data. The absolute configuration (2R, 4R) of 1 was determined via NOESY NMR and experimental and calculated ECD data analyses. Compounds 1–5 and 11 recently reported metabolites (hypoxoside, obtuside A, interjectin, acuminoside, curcapicycloside, and hypoxhemerolosides A – F) were screened for in vitro antimicrobial activity against various bacterial and fungal strains. Curcapicycloside and acuminoside exhibited antibacterial activity against Escherichia coli with 78 and 79% inhibition at 20 µg/mL, respectively. Hypoxhemeroloside A showed mild antifungal activity against Cryptococcus neoformans with 63% inhibition at 20 µg/mL.

Study was conducted In RCBD Design with three replicates to examine the impact of nitrogen application methods on the growth and yield of ridge gourd, evaluating 06 treatments comprising of various soil and foliar-applied N combinations. The following treatments were used: T1=Control (no N applied to the surface, no N applied to the leaves): T2=N at 75kg ha-1 through soil; T3=N at 75 kg ha-1 through soil+foliar urea at 1.5% concentration; T4=N at 75 kg ha-1 through soil+foliar urea at 3.0% concentration; T5=N at 100 kg ha-1 through soil; T6=N at100 kg ha-1 through soil+foliar urea at 1.5% concentration;T5=N at 100 kgha-1through soil+foliar urea at 1.5% concentration; The findings showe that various nitrogen application approaches had a major impact on all of the ridge gourd parameters calculated in this analysis (P0.05). With i258.33cm ivine iduration, 7.75 branchesivine-1, 11.26 fruits vine-1,i281.38giaverageisingle-fruitiweight, 3.25kg weightiofiridgeigourd fruitiperivine, 36.30kg ridge gourd fruit yield plot-1 and 14520.55kg ridge gourdifruit yieldiha-1, the ridge gourd vines plantediunderiT6 (N at 100kg ha-1 by soil+foliariapplicationiofiurea at 1.5%iconcentration)iratediforibothivine growthiand ridge igourd yield characteris-tics, the ridgeigourd cropifertilisediwith N at 100k ha-1 through oiliranked second, N at 75kg ha-1 through soil + foliar application ofiureaiati3.0%iconcentrationiranked third, N at 75kg ha-1 through the soil+foliar application of ureaiat 1.5% concentration ranked fourth, and N at 75 kg ha-1 through soil without foliariurea ranked fifth. Onialliof the characteristics tested, the controliplots came inilast.

In this study, the antifungal, antioxidant, enzyme inhibitory (α-amylase), phytochemical and cytotoxicity profile of Jatropha curcas seed oil and de-oiled seed cake extracts was determined. The antifungal activities were investigated against different phytopathogenic fungi including Colletotrichum coccodes, Pythium ultimum, Phytophthora capsici, Rhizoctonia solani, Bipolaris oryzae and Fusarium fujikuroi. Methanolic extract was found to be highly fungicidal as compared to aqueous extract, n-hexane extract and seed oil, against P. capsici and F. fujikuroi. Methanolic extract was also found to possess significantly higher antioxidant potential (p

Antimicrobial activities of deoiled seed kernel (mechanically pressed), fruit coat and seed coat of Jatropha curcas Linn. collected from two regions (Bannu and Peshawar) of Pakistan were investigated. The antimicrobial activities were carried out against Klebsiella pneumoniae (ATCC 43816), Escherichia coli (ATCC 10536), Staphylococcus aureus (ATCC 6538), Bacillus subtilis (ATCC 6633) and two clinical fungal isolates Aspergillus fumigatus and Candida albicans using agar well diffusion method. The antibacterial activities of Peshawar sample were found to be higher than Bannu, against selected strains. While antifungal activities of both samples were similar. Highest zone of inhibition 31.5 ± 0.7 mm was exhibited by n-hexane extract of deoiled seed kernel of Peshawar sample against Bacillus subtilis (ATCC 6633). The minimum inhibitory concentration of ethanolic extracts of deoiled seed kernel and seed coat of Peshawar sample was 31.25 - 25 mg/ml. Whereas, minimum inhibitory concentration of ethanolic and n-hexane extracts of Bannu sample was 62.5 - 125 mg/ml. The results suggested that antimicrobial potential of J. curcas Linn. varied with geographical distribution. The investigation of different varieties of medicinal plants belonging to the same species will greatly enhance the chances of best pharmaceuticals discovery. Bangladesh J. Bot. 50(2): 219-226, 2021 (June)

Background: Genus Berberis (family Berberidaceae), which contains about 650 species and 17 genera worldwide, has been used in folklore and various traditional medicine systems. Berberis Linn. is the most established group among genera with around 450-500 species across the world. This comprehensive review will not only help researchers for further evaluation but also provide substantial information for future exploitation of species to develop novel herbal formulations. Objective: The present review is focussed to summarize and collect the updated review of information of Genus Berberis species reported to date regarding their ethnomedicinal information, chemical constituents, traditional/folklore use, and reported pharmacological activities on more than 40 species of Berberis. Conclusion: A comprehensive survey of the literature reveals that various species of the genus possess various phytoconstituents mainly alkaloids, flavonoid based compounds isolated from different parts of a plant with a wide range of pharmacological activities. So far, many pharmacological activities like anti-cancer, anti-hyperlipidemic, hepatoprotective, immunomodulatory, anti-inflammatory both in vitro & in vivo and clinical study of different extracts/isolated compounds of different species of Berberis have been reported, proving their importance as a medicinal plant and claiming their traditional use.

Bacteria and fungi have a high potential to produce compounds that display large structural change and diversity, thus displaying an extensive range of biological activities. Secondary metabolism or specialized metabolism is a term for pathways and small molecule products of metabolism that are not mandatory for the subsistence of the organism but improve and control their phenotype. Their interesting biological activities have occasioned their application in the fields of agriculture, food, and pharmaceuticals. Metabolic engineering is a powerful approach to improve access to these treasured molecules or to rationally engineer new ones. A thorough overview of engineering methods in secondary metabolism is presented, both in heterologous and epigenetic modification. Engineering methods to modify the structure of some secondary metabolite classes in their host are also intensively assessed.

Mirabilis multiflora is an acclaimed hallucinogen consumed traditionally by the Hopi Indians to induce diagnostic visions. Its root extract afforded a new (3) and four known (2, 5, 6, and 7) 12a-hydroxyrotenoids, a known rotenoid (4), and two known secondary metabolites (1 and 8). The structures of the compounds were elucidated based on spectroscopic and spectrometric data analysis. Electronic circular dichroism data were used to define the (6aS,12aR) absolute configuration of the 12a-hydroxyrotenoids. Compounds 2-7 were screened for their radioligand binding affinities toward the opioid (δ, κ, and μ) and cannabinoid (CB1 and CB2) receptor subtypes. The 6-methoxy-substituted rotenoids 3, 4, and 7 showed the highest receptor binding affinity with moderate selectivity toward the δ-opioid receptor subtype, with negligible binding affinities for CB1 and CB2. Their binding affinities toward the δ-opioid receptor were 64.5% (4), 58.7% (7), and 55.3% (3) at 10 μM, respectively.

Aim: Candiduria is very common in hospitalized patients. It poses a clinical challenge for the physicians since it is usually asymptomatic. The aim of this study was to identify risk factors associated with nosocomial candiduria in urinary tract infection (UTI) suspected patients in Methodology: Intensive Care Unit (ICU) and to determine their antifungal sensitivity profile. The urine specimens (168) were collected, microscopically screened for presence of yeast, cultured and analyzed for counting, isolation, phenotypic identification of Candida albicans. and testing antifungal resistance profile. Data regarding age, gender, use of catheter, use of antibiotics, diabetes mellitus among patients was also recorded. Results: Out of 168 specimens, C. albicans were isolated from 69 specimens, whereas 20 specimens showed other Candida spp. Age >45 years, gender female, previous use of antibiotics, urinary catheterization, stay in ICU >1 week were found the main predisposing factors (p

In an attempt to find species specific markers, a phenoxychromone (1) and eight isoflavonoids including six isoflavans (2-7) and two isoflavanones (8 and 9) were isolated from the root of Glycyrrhiza uralensis. Among the isolated phenolic compounds, glycyurelone (1), glycyurelvanins A and B (2 and 3) were found to be undescribed while others, (-)-vestitol (4), conferol A (5), glyasperin C (6), glyasperin D (7), (-)-licoisoflavanone (8), and (-)-3'-(γ,γ-dimethylallyl)kievitone (9) were previously reported. All compounds except 4 and 5 were prenylated and majority of these possess isoflavan scaffold with highly conserved stereo specificity at C-3 center. Structure elucidation was mainly based on extensive NMR, ECD and mass spectral data analysis.

This study aimed at investigating the physicochemical and bread‐making features of dehydrated spinach. Physicochemical composition of spinach powder was compared with wheat flour and the effect of spinach powder supplementation on the nutritional composition, dough rheology, and quality attributes of chapatti were assessed. The results suggested spinach powder to be holding 8.2% crude fiber, 19.2% protein, 1,304 mg/100g calcium, and 40.4 mg/100g iron. Spinach powder indicated significantly increased values for hygroscopicity, swelling power, and water solubility index values, that is, 6.4%, 7.1 g/g, and 4.2%, respectively, when compared with wheat flour. Supplementation of spinach powder in wheat flour at 20% substitution level significantly reduced dough development properties including water absorption, dough stability, and peak dough development time. Color measurements of baked chapatti indicated a significant reduction in L*, a*, and chroma values with increasing the level of spinach powder supplementation; however, sensory profiling confirmed that supplementation of spinach powder at 7.5% had an optimum effect on the overall acceptability of the baked product. The results further suggested that replacing wheat flour with spinach powder (5%–7.5%, w/w) in baked products could be a viable dietary approach to enhance the optimum supply of micronutrients and to combat micronutrient deficiencies among various population segments., Our findings conclude that spinach powder supplementation at 7.5% could be a viable and practical approach to develop value‐added baked goods. The results further indicate that the utilization of green leafy vegetables as powder formulae or premixes could help abridge nutritional inadequacies and food insecurities cost‐effectively among vulnerable population groups.

The point of this study to investigate the impact of sports support on social improvement at an optional level in the region Muzaffarabad. The goals of the investigation were to discover the effect of sports interest on social improvement of the understudies (enrolled students) at the auxiliary level in District Muzaffarabad, Azad Jammu and Kashmir and to look at the social improvement of male and female understudies partaking in sports. The population of the study was all competitors while 444 of them were taken as a sample. The test of the examination comprised of 444 competitor understudies from chosen public area auxiliary schools of region Muzaffarabad through purposive inspecting. A self-created poll comprised of 35 shut finished inquiries was utilized as information Honesty estimated on a five-point Likert scale. The study was delimited to auxiliary schools of Muzaffarabad. The outcome demonstrated that there is no contrast between the social advancement of male and female competitor understudies at auxiliary school level in area Muzaffarabad. Noteworthy effect of sports support was found on social advancement in athletes' i.e. Self-conduct, friendship, social-conduct, building certainty and honesty at auxiliary level in region Muzaffarabad

Drought stress at the germination stage is an important environmental stress limiting crop yield. Hence, our study investigated comparative root transcriptome profiles of four contrasting soybean genotypes viz., drought-tolerant (PI342618B/DTP and A214/DTL) and drought-sensitive (NN86-4/DSP and A195/DSL) under drought stress using RNA-Seq approach. A total of 4850 and 6272 differentially expressed genes (DEGs) were identified in tolerant (DTP and DTL) and sensitive (DSP and DSL) genotypes, respectively. Principle component analysis (PCA) and correlation analysis revealed higher correlation between DTP and DTL. Both gene ontology (GO) and MapMan analyses showed that the drought response was enriched in DEGs associated with water and auxin transport, cell wall/membrane, antioxidant activity, catalytic activity, secondary metabolism, signaling and transcription factor (TF) activities. Out of 981 DEGs screened from above terms, only 547 showed consistent opposite expression between contrasting genotypes. Twenty-eight DEGs of 547 were located on Chr.08 rich in QTLs and "Hotspot regions" associated with drought stress, and eight of them showed non-synonymous single nucleotide polymorphism. Hence, 10 genes (including above eight genes plus two hub genes) were predicated as possible candidates regulating drought tolerance, which needs further functional validation. Overall, the transcriptome profiling provided in-depth understanding about the genetic mechanism and candidate genes underlying drought tolerance in soybean.

Four undescribed cembranoids, sarcoroseolides A-D (1-4) along with nine reported related cembranoids (5-13) were isolated from the soft coral Sarcophyton roseum. The chemical structures of sarcoroseolides A-D were elucidated by extensive 1 D and 2 D NMR as well as HR-ESIMS spectroscopic data. Moreover, the geometric and absolute configurations were assigned by the modified Mosher's method and/or NOESY experiments. The extract and the isolated metabolites were evaluated for their potential anti-inflammatory and cytotoxic activities.

Bioremediation of soil contaminated with organic chemicals is a challenging problem in the environmental scenario. On the basis of identification of remediation capability of Helianthus annus(sunflower) assisted with Pseudomonas aeruginosaand Micrococcus luteuswere used to investigate the soil contaminated with petroleum hydrocarbons (crude oil) at varying concentrations (25,50,75,100,125, &150ml/ kg of soil) in a pot experiment carried outin a green house. According to both qualitative and quantitative analysis of parameters including plant height, stem girth, number of leaves, leaf area and photosynthetic pigments including chlorophyll a, b and carotenoid indicated that H.annusshowed peculiar tolerance to the higher concentration of crude oil concentration in the soil containing both micrococcus and pseudomonas (MP) (combined) consortium showing positive synergetic effect on H.annusin remediating the petroleum hydrocarbons at all concentrations more significantly at higher concentration of crude oil (150ml/kg) of soil. The results show significant increase in growth of (both Micrococcus and Pseudomonas) MP treated plant as compare to control plants; in control the leaf area 18.716 ± 1.256, stem girth 3.40±0.10 and plant height 90.72±4.062 whereas in MP treated plant leaf area 19.65 ±0.292, stem girth 3.580 ±0.048, height 98.20±3.852 respectively. On the whole Helianthus anuusis commercial oil yielding crop that can be effectively appliedto phytoremediation of petroleum hydrocarbons assisted with Micrococcusand Pseudomonasis a significant treatment for remediating the soil contaminated with petroleum hydrocarbon.

Information is needed for comparative assessment and agronomic practices for phytoavoidation in multi-pollutant field. A field study was conducted to explore 97 Brassica pekinensis L. genotypes with permissible limit of contaminants growing in a severely Cd, moderately nitrate and slightly Pb multi-polluted field. Thirteen genotypes, i.e. KGZY, CXQW, CAIB, JINL, JQIN, JFEN, WMQF, XLSH, TAIK, BJXS, JUKA, XYJQ and GQBW, were identified with permissible limit for nitrate, Cd and Pb based on their resistance to heavy metal and nitrate accumulation in leaves when grown in co-contaminated soils. Furthermore, the correlation between essential and toxic elements concentrations in plant of B. pekinensis were inconsistent. Generally speaking, application of increasing Ca, K and S fertilizers in appropriate forms and dosages tended to increase the yield and quality of B. pekinensis cultivated in multi-pollutant field.

Glycyrrhiza inflata Batalin is among the three glycyrrhizin producing Glycyrrhiza species and can be distinguished from other species with regard to its retrochalcone contents. Seven retrochalcones, echinatin and licochalcones A, C, D, E, K, and L were isolated and characterized from the chloroform extract of G. inflata roots. Among the isolates, licochalcone L was found to be previously undescribed. Structure elucidation of these specialised metabolites was achieved through NMR and mass spectroscopic data analyses.

A rare dimeric sesquiterpenoid (tinosposinoside, 1) and a phenylpropanoid (cordifolioside C, 2), two undescribed metabolites, were isolated from the methanolic extract of the stems of Tinospora sin...

Phytochemical investigation of the methanolic extract of Vangueria agrestis aerial parts led to the isolation and characterization of four monoterpene rhamnosides, two ursane saponins, and six flavonoid glycosides. Of these constituents, are three undescribed monoterpene glycosides, vangagrestosides A-C. The reported constituents comprise the monoterpene glycoside [(2E,6Z)-2,6-dimethyl-8-[O-α-L-rhamnopyranosyl-(1→3)-α-L-rhamnopyranosyl)-oxy]-octadien-1-yl α-L-rhamnopyranoside], the saponins [3-O-[α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl]pomolic acid, and 3-O-[α-L-rhamnopyranosyl-(1→2)-β-D-xylopyranosyl]pomolic acid], and the flavonoids [narcissoside, rutin, hyperoside, guajavarin, biorobin, and nicotifloroside].

Herein, synthesis of rhamnolipid surfactants was performed using a non-pathogenic Pseudomona s putida strain cultured on a variety of waste frying oils (WFOs) under Taguchi multi-objective optimization design. The effect of substrate types, fermentation setups and incubation time on the biomass concentration, rhamnolipid yield and surface tension of the cultivate media has been investigated. The results demonstrate that the multi-objectives investigation helps to document the optimal limits of the process parameters based on Gray relational analysis. After finding the optimal conditions, a validati on run was performed; therein, the rhamnolipid yield increased from 3.4 to 4.1 g/L; the biomass concentration decreased by 4.84% with an additional surface tension reduction of 2.19% due to an increase of rhamnolipids yield. Overall, soybean WFO was observed to be a preferred substrate for P. putida strain both under Taguchi design and the validation run. The present study proposes a low total of runs and optimum product yield under the Taguchi based multi-objective optimization.

Pink bollworm, Pectinophora gossypiella (Saunders), (Lepidoptera: Gelechidae) is considered as a notorious and destructive pest in the world and can reduce cotton yield up to 30% if not managed properly. Insect pest management through mating disruption technique offers practically an ideal approach to combat the pests. In the present study, the efficacy of mating disruption dispensers (PB rope L) for the management of pink bollworm was evaluated and the percent infestation in bolls and leftover bolls was determined in the cotton growing areas of Punjab, Pakistan during 2017. The PB ropes @100 per acre were used for each treated field of approximately 50 acres with a block of 10 acres as a control plot in the same union council of the treated plot. Calculation was made by comparing infestation and cotton yield in treated field with that of control field. The PB rope significantly reduced both the number of males in the sex pheromone traps and cotton bolls damage. Results revealed that PB rope during the first pin square period (45 days after sowing) effectively minimized the damage even in leftover bolls. The yield of treated field was enhanced compared with control field by 31% in 2017. Infestation in leftover bolls was estimated by comparing cotton bolls collected from control field with bolls from the treated field. The results reflected 1.56% damage in control field and 0.30% damage in treated field with a difference of 1.26 (80% reduction in damage in treated plots). Result indicated that PB rope disrupted the pest mating effectively during July to October.

In recent decades, Pakistan has suffered a decline in cotton production due to several factors, including insect pests, cotton leaf curl disease (CLCuD), and multiple abiotic stresses. CLCuD is a highly damaging plant disease that seriously limits cotton production in Pakistan. Recently, genome editing through CRISPR/Cas9 has revolutionized plant biology, especially to develop immunity in plants against viral diseases. Here we demonstrate multiplex CRISPR/Cas-mediated genome editing against CLCuD using transient transformation in N. benthamiana plants and cotton seedlings. The genomic sequences of cotton leaf curl viruses (CLCuVs) were obtained from NCBI and the guide RNA (gRNA) were designed to target three regions in the viral genome using CRISPR MultiTargeter. The gRNAs were cloned in pHSE401/pKSE401 containing Cas9 and confirmed through colony PCR, restriction analysis, and sequencing. Confirmed constructs were moved into Agrobacterium and subsequently used for transformation. Agroinfilteration in N. benthamiana revealed delayed symptoms (3–5 days) with improved resistance against CLCuD. In addition, viral titer was also low (20–40%) in infected plants co-infiltrated with Cas9-gRNA, compared to control plants (infected with virus only). Similar results were obtained in cotton seedlings. The results of transient expression in N. benthamiana and cotton seedlings demonstrate the potential of multiplex CRISPR/Cas to develop resistance against CLCuD. Five transgenic plants developed from three experiments showed resistance (60−70%) to CLCuV, out of which two were selected best during evaluation and screening. The technology will help breeding CLCuD-resistant cotton varieties for sustainable cotton production.

At present time, every nation is absolutely concern about increase agricultural production and bioremediation of petroleum contaminated soil. Hence, with this intention in current study potent natural surfactant (surfactin) was evaluated for low-cost production by Bacillus subtilis SNW3, previously isolated from Fimkessar oil field, Chakwal Pakistan. The best results were obtained using substrates in combination (white beans powder (6% w/v) plus waste frying oil (1.5% w/v) and (0.1% w/v) urea) with surfactin production of about 1.17 g/L contributing 99% reduction in cost required for medium preparation. To the best of our knowledge, no single report is present describing surfactin production by Bacillus subtilis using white beans powder as a culture medium. Surfactin was confirmed as the principal product characterized by thin-layer chromatography (TLC) and Fourier-transform infrared spectroscopy (FTIR). Additionally, produced surfactin display great physicochemical properties of surface tension reduction value (SFT=28.8 mN/m), significant oil displacement activity (ODA=4.9 cm), excessive emulsification ability (E24=69.8 %), and attains critical micelle concentration (CMC) value at 0.58 mg/mL. Furthermore, surfactin exhibits excellent stability over an extensive range of pH (1-11), salinity (1-8%), temperature (20-121°C) and even after autoclaving. Subsequently, surfactin produced proved suitable for bioremediation of crude oil (86%) and as potent plant growth-promoting agent that significantly (P

In this study, Rhizobium rhizogenes-mediated root proliferation system in Sedum alfredii has been established. Twenty strains of R. rhizogenes were screened for root proliferation. A significant difference (P0.01) was observed in plant morphological characters under influence of different bacterial strains. The highest root fresh weight (3.236 g/plant) was observed with strain AS12556. Furthermore, significant difference (P0.05) was observed in the chemical composition of organic acids, Tartaric acid (TA), Succinic acid (SA), Malic acid (MA), Citric acid (CA) and Oxalic acid (OA), pH, Total Nitrogen (TN), Total Organic Carbon (TOC) and soluble sugars in root exudates with different R. rhizogenes mediated roots. Furthermore, a series of hydroponics experiments were conducted with varying concentrations of Cd (25, 50 and 75 µM) and Zn (100, 200 and 500 µM) to assess the phytoextraction efficiency of proliferated roots with Rhizobium. Several plants with proliferated roots showed enhanced growth and improved metal extraction efficiency. Five strains (LBA 9402, K599, AS12556, MSU440 and C58C1) were identified as potential strains for root proliferation in Sedum alfredii. R. rhizogenes strain AS12556 improved Cd/Zn phytoextraction by exogenous production of phytochemicals to promote root proliferation, improved shoot biomass, lowered oxidative damage and enhanced phytoextraction efficiency in S. alfredii. Therefore, it has been selected as a potential microbial partner of S. alfredii to develop extensive rooting system for better growth and enhanced phytoremediation potential. Results suggest that R. rhizogenes mediated root proliferation system can be used for optimizing metal extraction from contaminated soils.

Climate change is decreasing overall yield of crops through amplified water stress, increased disease, pests and weeds’ incidence, and shortened growing season. Sorghum [(Sorghum bicolor) L. Monch], a staple food of around 500 million individuals, promises to be the future crop to ensure food security as it has great adaptation against high temperature, high salinity, and drought stress. It is a C4 plant like many other grasses, that is, maize, sugarcane, millet, and switch grass. It can be used as an alternate energy crop, human food, livestock feed, and for medicinal and industrial purposes. The biomass of sorghum can be used as a source of carbohydrate or as a biofuel via thermochemical route. Although sorghum is relatively more adapted to different stresses, challenges seem to be different in the 21st century. In this chapter, we have summarized the literature related to different biotic and abiotic stresses that are threating sorghum productivity and also discussed how sorghum plant responds to these climatic factors. We have also summarized the use of different conventional as well as modern breeding tools that will help in ideotyping the model sorghum plants to cope future challenges and ensure food security.

Anthemis austriaca Jacq. flowers are traditionally used to alleviate abdominal pain, hemorrhoids, ovary diseases and pneumonia. This study aimed to investigate the effects of A. austriaca flowers, which are frequently used in gynecological disorders, on the rat endometriosis model. The rat endometriosis model was used to evaluate the potential activity of the plant in endometriosis. The dried plant material was extracted with n-hexane, ethyl acetate (EtOAc), and methanol (MeOH), successively. The obtained extracts from A. austriaca flowers were applied to the rats. The adhesion scores, endometrial foci areas, and cytokine levels of the peritoneal fluids were measured on surgical induction of endometriosis in rats. The adhesion scores, endometriotic volume, and cytokine levels of the peritoneal fluids were reduced in the EtOAc, MeOH, and buserelin acetate-treated (reference) groups. The MeOH extract reduced the adhesion scores and endometrial foci areas from 3.1 to 1.1 (p < 0.01) and from 86.4 to 40.5 (p < 0.01), respectively and also the MeOH extract reduced tumor necrosis factor (TNF)-alpha, vascular endothelial growth factor (VEGF), and interleukin (IL)-6 levels of the peritoneal fluids from 13.7 to 3.8 (p < 0.01), from 28.4 to 16.3 (p < 0.05) and from 50.2 to 24.3 (p < 0.01), respectively. Therefore, isolation studies were conducted on the EtOAc and MeOH extracts. After the MeOH extract was fractionated using RP-18 column, the obtained subfractions were evaluated again on the endometriosis rat model. Subfractions A and C of the MeOH extract displayed statistically significant activity on the endometriosis rat model. Phytochemical investigation resulted in the isolation of 4-beta-D glucopyranosyloxy-6-methyl-2H-pyran-2-one (1) from Fr. A and quercetin (2), apigenin-7-O-(3 ''-O-acetyl)-beta-D-glucopyranoside (3), apigenin-7-O-(6 ''-O-acetyl)-beta-D-glucopyranoside (4), apigenin-7-O-beta-D-glucopyranoside (5), quercetin-7-O-beta-D-glucopyranoside (6) from Fr. C. Moreover, beta-sitosterol-3-O-beta-D-glucopyranoside (7) was isolated from the EtOAc extract. As a conclusion, the MeOH extract obtained from A. austriaca flowers contributed to the regression of endometriosis. In addition, flavonoids and sterols of the plant were detected as the possible compounds responsible for the activity. (C) 2019 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.

Conocarpus lancifolius is a fast-growing and drought tolerant tree species with phytoremediation potential in arid environments. The present study was conducted to evaluate the phytoaccumul...

Sedum alfredii Hance is a hyperaccumulator of cadmium (Cd) and zinc (Zn) that exhibits extraordinary accumulation of these metals in various tissues. The Nicotianamine synthase (NAS) genes play key roles in regulating the production of nicotianamine, a non-protein amino acid that facilitates metal homeostasis in plants, but the functions of these genes in S. alfredii remain unknown. The aims of this study were to identify and characterize the NAS gene from S. alfredii (SaNAS1) and explore NAS roles in Cd or Zn tolerance and accumulation. One Cd-induced NAS gene from S. alfredii (SaNAS1) was cloned. We determined the subcellular localization of the encoded protein and evaluated SaNAS1 expression. We also assessed SaNAS1 function by conducting a complementary assay with yeast mutants Δzrc1 exposed to Cd or Zn. Finally, transgenic Arabidopsis plants expressing SaNAS1 were produced, and SaNAS1 function was further examined in these plants. SaNAS1 was highly expressed in response to Cd or Zn exposure, and the encoded protein was distributed throughout the cytoplasm and nucleus. Furthermore, yeast expressing SaNAS1 exhibited increased tolerance to Cd or Zn. Finally, expression of SaNAS1 in Arabidopsis increased nicotianamine production and promoted Cd or Zn accumulation in roots and shoots. SaNAS1-expressing transgenic Arabidopsis lines showed improved seedling growth under Cd or Zn stress compared with wild type, indicating that SaNAS1 enhances tolerance to both Cd and Zn. SaNAS1 may play a critical role in Cd or Zn tolerance and hyperaccumulation by regulating nicotianamine level in S. alfredii.

The rhizomes of Bulbine natalensis furnished six previously unreported anthraquinone derivatives (1-6), together with eight known specialized metabolites. Their structures were determined by interpretation of 1D and 2D NMR and HRESIMS data. The absolute configurations of compounds 1-6 were determined by specific rotation and circular dichroism experiments. The isolated compounds were evaluated for antimicrobial activities, and compound 1 was found to be a moderate inhibitor (IC50 0.02 μM) against methicillin-resistant Staphylococcus aureus (MRSA).

Sceletorines A and B, two unique alkaloids, were isolated for the first time from the aerial parts of Mesembryanthemum tortuosum. Their structures were determined by the extensive spectroscopic methods and their absolute configuration was established with ECD calculations. Plausible biosynthetic pathways for 1 and 2 are proposed and a relationship to the previously assumed artifact, channaine is also addressed.

Spinach (Spinacia oleracea L.) is an important leafy vegetable, rich in various essential nutrients required for human health. However, it may also accumulate hazardous materials in edible parts, and subsequently poses human health risk. Evaluation of variation in essential nutrients and hazardous materials in spinach genotypes is important for breeding desired genotypes for human consumption. In this study, we investigated 68 spinach genotypes for screening desired genotypes. Seven spinach genotypes, i. e. KBXX, XIBW, KBWZ, DBSC, LLCJ, QUAN and JXQD were identified as safe genotypes that contained contaminants in edible parts below permissible limit (Cd

Comparative impact of CO2 application and endophyte inoculation was investigated on the growth, rhizosphere characteristics, and cadmium (Cd) absorption of two ecotypes of Sedum alfredii Hance in response to Cd stress under hydroponic or rhizo-box culture conditions. The results showed that both CO2 application and endophyte inoculation significantly (P

Introduction: Bunium persicum (Boiss.) B. Fedtsch belongs to family Apiaceae of genus Bunium, which is comprised of about 166 species and is widely distributed in Kashmir valley. It is used medicinally in dyspepsia and diarrhea, digestive and urinay tract infections, chronic gastiritis, colitis, anticonvulsant, antiasthma and diuretic. Objectives: The current study deals with Pharmacognostic parameters of the fruits of Bunium persicum (Boiss.) B. Fedtsch. Materials and Methods: The fruits of Bunium persicum (Boiss.) B. Fedtsch was collected, shade dried for about 2 weeks and powdered and the powered fruit part of plant material was evaluated for Pharmacognostic parameters by standard methods. The hydroalcholic extract of Bunium persicum was subjected to preliminary phytochemical screening for the presence of various Phytoconstituents. The microscopy of the fruits of Bunium persicum (Boiss.) B. Fedtsch reveals the presence of Part of a group of sclereids from the mesocarp, thicker-walled sclereids with adjacent thin walled parenchyma, Fragment of vitta, Thin-walled fibre and Calcium-oxalate crystals. Results: Proximate analysis of the fruits of Bunium persicum (Boiss.) B. Fedtsch showed that the dried fruit powder has 34.0 % Total ash value, 29.0 % Acid insoluble ash value, 9.5 % Sulphated ash value. Loss on drying was found to be 6.2 %. The hydroalcholic extract of Bunium persicum was found to contain various phytoconstituents. Fluorescence analysis of the fruit powder showed the behavour, when treated with different chemical reagents. Conclusion: The current study displayed the microscopical characters, the preliminary phytochemical screening and the proximate analysis of the fruits of Bunium persicum (Boiss.) B. Fedtsch. Data composed from such studies can be used as standard in the quality control of this plant as a herbal medicine for treatment of various diseases.

Rooibos tea ( Aspalathus linearis) is a well-known South African herbal tea enjoyed worldwide. Limited reports indicate the potential of rooibos tea to alter the activity of certain cytochrome P450 (CYP450) isozymes. In this study, the phytochemical investigation of MeOH extract of A. linearis (leaves and stems) resulted in the isolation and characterization of 11 phenolic compounds. The MeOH extract exhibited significant inhibition of the major human CYP450 isozymes (CYP3A4, CYP1A2, CYP2D6, CYP2C9, and CYP2C19). The strongest inhibition was observed by the extract for CYP3A4 (IC50 1.7 ± 0.1 μg/mL) followed by CYP2C19 (IC50 4.0 ± 0.3 μg/mL). Among the tested phytochemicals, the most potent inhibitors were isovitexin on CYP3A4 (IC50 3.4 ± 0.2 μM), vitexin on CYP2C9 (IC50 8.0 ± 0.2 μM), and thermopsoside on CYP2C19 (IC50 9.5 ± 0.2 μM). The two major, structurally related compounds aspalathin and nothofagin exhibited a moderate pregnane-X receptor (PXR) activation, which was associated with increased mRNA expression of CYP3A4 and CYP1A2, respectively. These results indicate that a high intake of nutraceuticals containing rooibos extracts may pose a risk of herb-drug interactions when consumed concomitantly with clinical drugs that are substrates of CYP enzymes.

Phytochemical study on the stem bark of Clausena excavata Burm. f. led to the isolation and identification of 11 compounds, including four prenylated coumarins (1–4), three carbazole alkaloids (5–7), a limonoid (8), a sterol (9), a depside (10) and a xanthone (11). The chemical structures of the compounds were elucidated based on NMR and mass spectroscopic data, and further supported by comparison with previously literature values. This is the first report on the isolation of compounds 10 and 11 from the genus Clausena. The chemotaxonomic significance of the isolated compounds is discussed.

Background: Burn injuries are common and major health problem throughout the world. The burn wound represents as a favorable area for opportunistic colonization of microorganisms with exogenous and endogenous origin. In burns patients infections arise from multiple sources. Burn wounds become initially colonized and infected with Gram positive bacteria, mainly Staphylococci, that are superseded during the second week by Gram Negative bacteria. Objectives: it is a microbial surveillance retrospective study; that aimed to evaluate the significance of Rule of nine in diagnosis of aerobic bacterial burn wound infection and carried out in between June 2007 to September 2011 in the Department of Pharmaceutics, University of Karachi. Study Design: Descriptive retrospective study. Materials: A total of 118 patient irrespective of age, sex, date and time of burn, date and time of hospital admission, interval between time of burn and hospital admission, degree and percentage (%) of burn and duration of hospital stay (when specimen collected) were registered for this study. All patients were divided into two groups (A and B). Results: Out of 58 patients of group A isolation rate per patient was 1.1 while in group B it becomes 3.0. According to TBSA the isolation rate in group B rises with rise in TBSA. Most prevalent organism in these patients was found S. aureus(23%) P.aeruginosa(21%). Conclusions: Burn patients are incubator for variety of aerobic bacteria and rate of isolation of these organisms increase with rise in TBSA. The wounds of these patient must required continuous microbial surveillance that may reduce the rate of mortality

Objective: To determine the frequency of dyslipidemia in Helicobacter pyloriinfected patients. Patients and methods: This cross sectional descriptive study of six monthsstudy was conducted at Liaquat University Hospital Hyderabad from 01-03-2012 to 31-08-2012. All the patients between 19 to 60 years of age present with symptoms of dyspepsia,bloating or epigastric discomfort for more than 01 week duration were admitted and evaluatedfor Helicobacter pylori infection. Thereafter the positive cases (Helicobacter pylori infectedpatients) were further evaluated for dyslipidemia. Results: During six month study period,total 144 patients (95 males and 49 females) with Helicobacter pylori infection were evaluatedfor dyslipidemia (lipid profile). Majority of patients were from urban areas 110/144 (76%).The mean ±SD for age of patients with Helicobacter pylori infection was 35.94±10.77. Themean age ±SD of dyslipidemic patient was 32.62±6.52. The dyslipidemia was identified in 87(60.4%) patients, of which 51(58.6%) were males and 36(41.4%) were females. Regarding thepattern of dyslipidemia, ten (11.4%) patients had raised serum triglycerides level, twenty eight(32%) had raised LDL-C level, twenty six (28.9%) had raised serum cholesterol level, seven(8.7%) had low HDL-C level and sixteen (18.3%) had mixed dyslipidemia. The mean ±SD ofhypertriglyceridemia, ↑HDL-C, ↑LDL-C and hypercholesterolemia in dyslipidemic Helicobacterpylori infected was 280.72±22.85, 24.21±2.63, 180.63±12.98 and 285.21±23.63 respectively.Conclusions: The H. pylori infected patients are prone to acquire dyslipidemia, therefore thepresent study observed 60.4% prevalence of dyslipidemia with male predominance (58.6%)

Background: Biosurfactants, being environment friendly, highly biodegradable, less toxic and stable compounds have applications in several environmental and industrial sectors that includes cosmetics, biomedical, bioremediation, and agriculture. Growing concern about eco-friendly compounds leads to replacement of chemical surfactants with biological surfactants. However, use of biosurfactant limits due to high production cost. Surfactin, a class of lipopeptide, considered as powerful biosurfactants having wide applications in therapeutics and environmental field. This study aims to investigate production and characterization of surfactin by Bacillus subtilis SNW3 and evaluating their potential application in seed germination and plant growth. Results: In present study, Bacillus subtilis SNW3 was previously isolated from Chakwal Pakistan and used for biosurfactant production. Optimized media for biosurfactant production was at (6% w/v) white beans powder in combination with (1.5% w/v) waste frying oil and (0.1% w/v) urea that shows surface tension reduction (28.8 mN/m), oil displacement assay (4.9 cm) and emulsification index (69.8 %). Environmental growth parameters like temperature (30 °C), inoculum size (1%), pH (6) and agitation (150 rpm) exhibit important role towards enhanced biosurfactant yield. Furthermore, surfactin obtained was found to be most stable at (5-7) pH, (8%) NaCl and (100 °C) temperature. Biosurfactant obtained was of lipopeptide nature called surfactin characterized by thin-layer chromatography (TLC) and fourier-transform infrared spectroscopy (FTIR). The surfactin obtained, used in a concentration of (0.7 g/100 mL) helps in seed germination and significantly enhanced growth of Solanum lycopersicum (tomato), Pisum sativum (pea), Capsicum annuum (peppers) and Lactuca sativa (lettuce).Conclusions: Bacillus subtilis SNW3 produces surfactin with more stability, that makes it useful for processing of food and in agriculture. The use of white beans powder and waste frying oil as sole source of carbon and energy makes the biosurfactant production more profitable, and environment friendly procedure by utilizing food processing by-products and wastes as substrate. Results obtained provide understanding about surfactin use for seed development and plant growth.

Airborne bioaerosols and particulate matter (PM) have been associated with asthma occurrence. Due to the adverse indoor environment and the absence of any baseline data for asthmatic patients of Pakistan, this study was aimed to establish a correlation between microflora and PM of residential microenvironments of asthmatic patients. This pilot study was conducted in different residential settings of asthmatic patients registered in the Jinnah hospital, Lahore. The characterization of PM (PM01, PM2.5, PM10) and bioaerosols were carried out in the houses of fifty patients that were categorized into four groups; A-large (418.06 m2 ), B-medium (211 m2 ), C-medium (104 m2 ), and D-small (62.71 m2 ) houses. The PM concentrations were monitored; using the DustTrack8533 aerosol monitor and the bioaerosols were characterized up to the Genus; using the culture-based method and biochemical testing. The bioaerosols were sampled; using the expose plate method and were analyzed using morphological features and biochemical tests. Eleven types of fungi and seven bacterial types were found in the air samples. The tendency of asthma occurrence is linked with higher Alternaria spp and Aspergillus spp. The mean indoor readings of PM01, PM2.5, PM10 were highest in D-category (331.75, 342.5, and 502.33 respectively). Moreover, the highest bacterial 9618 CFU/m3 ) and fungal levels (3092 CFU/m3 ) were also seen in D-category. According to two-way ANOVA, bacterial concentration was significantly different among the four groups while fungi concentration was non-significant (p

Background Bulbine natalensis Baker and Bulbine frutescens (L.) Willd., belonging to the family Asphodelaceae, are widely distributed in South Africa and traditionally used as an aphrodisiac and skin remedies. Objective The aim of this study is to develop an analytical method for chemical profiling and identification of components in Bulbine species, which would be useful for herbal identification and understanding of the biological activity of B. natalensis in terms of safety and benefits to human health. Method The anthraquinone-type compounds were structurally characterized from the extracts of dried stem and roots of Bulbine species and dietary supplements using liquid chromatography–quadrupole time-of-flight mass spectrometry (LC-QToF) with negative and positive ion electrospray. The calculated accurate masses of the protonated and deprotonated molecules and fragment ions were used for identification of the components from two Bulbine species. Results A total of 55 anthraquinone-type compounds, including 11 standard compounds, were identified in the crude extracts of two Bulbine species. Two Bulbine species and dietary supplements were clustered into different groups and possible chemical markers were identified. Conclusions The developed analytical method provided a fast and economic method for quality assessment of Bulbine species in dietary supplements based on anthraquinone-type compounds. Highlights This study reports holistic chemical profiling of Bulbine species using LC-QToF. The developed analytical method enabled non-targeted analysis of components in B. natalensis and B. frutescens, and is recommended for commercial and regulatory purposes.

Stem and leaf of Cissus quadrangularis L. (Vitaceae), indigenous to Asia and Africa, were used for medicinal and dietary purposes with limited information about the plant's phytochemistry. Stem and leaf samples were assessed for the simultaneous determination of polyphenolic compounds (catechin, epicatechin, quercetin-3-O-β-glucopyranoside, kaempferol-3-O-β-glucoside, quercetin-3-O-β-rhamnoside, leachianol F, amurensin A, pallidol, resveratrol, and quadrangularin A), using UHPLC-PDA-MS. The validation data showed that the method is precise, specific, accurate, and linear over the range of 0.5-100 μg/mL. Reversed-phase ultra-high performance liquid chromatography (UHPLC) fingerprints of the crude methanolic stem and leaf extracts of C. quadrangularis were obtained at different wavelengths based on their λmax. Polyphenolics were characterized using both UHPLC-PDA-MS and LC-QToF analysis. From liquid chromatography quadrupole time of flight-electrospray ionization mass spectrometry (LC-QToF) spectra, over 40 components were structurally correlated, and confirmation was based on the fragmentation characteristics and also from the information available in the literature. In addition to the LC-QToF method, a simple, fast HPTLC method was developed as a visual aid for the rapid qualitative analytical tool to help establish the quality assessment of botanical raw materials and dietary supplements claiming to contain Cissus.