Search

This study aims to establish a facile, user-friendly, accurate, and repeatable spectrophotometric detection method for Cr(III) ions in trace quantities. The method utilises reagent 1-nitroso-2-naphthol (NNPh) in a sodium dodecyl sulphate (SDS) 1.0 % micellar solution, replacing the old, time-consuming, and expensive solvent extraction method that employs hazardous solvents such as carbon tetrachloride and chloroform. SDS surfactant is used to solubilise indissoluble chelates, making the proposed technique easy, rapid, and environmentally friendly. The established method exhibits an improved linear calibration range, sensitivity, limit of detection, and selectivity compared to earlier spectrophotometric methods employing different complexing agents. The stoichiometric ratio between Cr and NNP was observed to be 1 : 3. The molar absorption coefficient (ε) was determined to be 2.05 ∙ 104 l mol−1 cm−1 at λmax 579 nm. The limits of detection for Cr(III) was measured as 3.49 ng l−1. A calibration linear curve was achieved in the concentration range of 0.5–5.0 μg ml−1. Results obtained showed good comparison with the results obtained by atomic absorption spectrometry. The developed method has been adapted for the analysis of Cr(III) in various alloys, pharmaceutical, environmental, and biological samples.

Abstract Tanacetum falconeri is a significant flowering plant that possesses cytotoxic, insecticidal, antibacterial, and phytotoxic properties. Its chemodiversity and bioactivities, however, have not been thoroughly investigated. In this work, several extracts from various parts of T. falconeri were assessed for their chemical profile, antioxidant activity, and potential for enzyme inhibition. The total phenolic contents of T. falconeri varied from 40.28 ± 0.47 mg GAE/g to 11.92 ± 0.22 mg GAE/g in various extracts, while flavonoid contents were found highest in TFFM (36.79 ± 0.36 mg QE/g extract) and lowest (11.08 ± 0.22 mg QE/g extract) in TFSC (chloroform extract of stem) in similar pattern as found in total phenolic contents. Highest DPPH inhibition was observed for TFFC (49.58 ± 0.11 mg TE/g extract) and TFSM (46.33 ± 0.10 mg TE/g extract), whereas, TFSM was also potentially active against (98.95 ± 0.57 mg TE/g) ABTS radical. In addition, TFSM was also most active in metal reducing assays: CUPRAC (151.76 ± 1.59 mg TE/g extract) and FRAP (101.30 ± 0.32 mg TE/g extract). In phosphomolybdenum assay, the highest activity was found for TFFE (1.71 ± 0.03 mg TE/g extract), TFSM (1.64 ± 0.035 mg TE/g extract), TFSH (1.60 ± 0.033 mg TE/g extract) and TFFH (1.58 ± 0.08 mg TE/g extract), while highest metal chelating activity was recorded for TFSH (25.93 ± 0.79 mg EDTAE/g extract), TFSE (22.90 ± 1.12 mg EDTAE/g extract) and TFSC (19.31 ± 0.50 mg EDTAE/g extract). In biological screening, all extracts had stronger inhibitory capacity against AChE while in case of BChE the chloroform extract of flower (TFFC) and stem (TFSC) showed the highest activities with inhibitory values of 2.57 ± 0.24 and 2.10 ± 0.18 respectively. Similarly, TFFC and TFSC had stronger inhibitory capacity (1.09 ± 0.015 and 1.08 ± 0.002 mmol ACAE/g extract) against α-Amylase and (0.50 ± 0.02 and 0.55 ± 0.02 mmol ACAE/g extract) α-Glucosidase. UHPLC-MS study of methanolic extract revealed the presence of 133 components including sterols, triterpenes, flavonoids, alkaloids, and coumarins. The total phenolic contents were substantially linked with all antioxidant assays in multivariate analysis. These findings were validated by docking investigations, which revealed that the selected compounds exhibited high binding free energy with the enzymes tested. Finally, it was found that T. falconeri is a viable industrial crop with potential use in the production of functional goods and nutraceuticals.

Background: Levofloxacin is a broad spectrum quinolone, widely used to treat infections caused by gram negative and gram positive bacteria. Development of resistance by pathogens against different broadspectrum antibiotics is increasing and now becoming a global issue. Objectives: The aim of the study is to evaluate the current sensitivity pattern of levofloxacin against various common clinical isolates like Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa,and Klebsiellapneumoniae. Methods: A total of one hundred and ten samples were collected from different pathological laboratories of Karachi, Pakistan. The above mentioned pathogens were isolated from blood, stool/urine, sputum, skin samples. Results: Results show least resistance of levofloxacin against E.coli (27.5%), and P.aeruginosa (27%), while S.aureus possessed highest resistance (45%). Conclusion: Study concluded levofloxacin still possesses excellent anti-microbial activity against common pathogens. Routine monitoring and surveillance is further required to ensure effective treatment regimens to community. KEY WORDS: Levofloxacin, Resistance, Susceptibility, Clinical Isolates, Pathogens.

Background: COVID-19 is primarily known as a respiratory illness; however, many patients present to hospital without respiratory symptoms. The association between non-respiratory presentations of COVID-19 and outcomes remains unclear. We investigated risk factors and clinical outcomes in patients with no respiratory symptoms (NRS) and respiratory symptoms (RS) at hospital admission. Methods: This study describes clinical features, physiological parameters, and outcomes of hospitalised COVID-19 patients, stratified by the presence or absence of respiratory symptoms at hospital admission. RS patients had one or more of: cough, shortness of breath, sore throat, runny nose or wheezing; while NRS patients did not. Results: Of 178,640 patients in the study, 86.4 % presented with RS, while 13.6 % had NRS. NRS patients were older (median age: NRS: 74 vs RS: 65) and less likely to be admitted to the ICU (NRS: 36.7 % vs RS: 37.5 %). NRS patients had a higher crude in-hospital case-fatality ratio (NRS 41.1 % vs. RS 32.0 %), but a lower risk of death after adjusting for confounders (HR 0.88 [0.83–0.93]). Conclusion: Approximately one in seven COVID-19 patients presented at hospital admission without respiratory symptoms. These patients were older, had lower ICU admission rates, and had a lower risk of in-hospital mortality after adjusting for confounders.

In this study, we solved the Schrödinger wave equation by using effective potential in an artificial neural network (ANN) for the mass spectrum of different charmonium states, including ηc, ψ2, χ2, and χ4. The ANN approach provides an efficient, more general, and continuous solution-approximating strategy, thus eliminating the possibility of skipping any region of interest in mass spectroscopy. The close consistency of ANN results with the already-reported results from numerical and theoretical approaches and experimental ones shows the reliability and accuracy of the ANN approach.

Objective: To concisely compare genomic profiles of XDR Salmonella Typhi isolates from Lahore with antimicrobial-resistant Salmonella Typhi isolates from other developing nations. Study Design: Comparative analysis of whole genome sequences. Place and Duration of Study: The study was conducted from December 2020 to September 2021 at Kauser Abdulla Malik (KAM) School Life Sciences, Forman Christian College University, Lahore, Pakistan. Methods: The Galaxy pipeline was run to obtain detailed information at the nucleotide level regarding mutations that lead to the emergence of XDR strains. Whole genome sequences were analyzed to compare the genomes of selected three developing nations. Results: The Pakistani isolates had a significantly higher mutation rate, higher proportion of modifiers, and silent mutations as compared to isolates of Bangladesh and India. Conclusion: Cases of Salmonella Typhi XDR are rapidly rising in Asian countries such as Pakistan, Bangladesh, and India emphasizing the need to analyze and compare its genome with relevant strains. Our study highlights the unique profile of the Lahore (Pakistan) isolate with the highest mutation rate suggesting the potential regional differences in selective pressure. Further spotlights the necessity to elucidate the functional consequences of the identified mutations in S. Typhi isolates. How to cite this: Niaz M, Mumtaz Z, Ahmed M, Ahmad S, Ali A, Yousaf MZ. Variant Analysis of XDR Salmonella Typhi Strains Using Global Alignment Tool Kit in South Asian Region. Life and Science. 2024; 5(2): 244-250. doi: http://doi.org/10.37185/LnS.1.1.432

Abstract A comprehensive study was carried out on in vitro multiplication and rooting using the medium enriched with different plant growth regulators and acclimatization of pineapple cv. ‘Smooth Cayenne’ using different soil growing substrates. The significantly highest shoot buds (Avg. 16.7) were obtained on the medium comprising 2.0 mg L−1 BA (6-Benzylaminopurine). Results showed that 1.0 mg L−1 IBA (Indole-3-butyric acid) increased the thickness and length of white adventitious roots and resulted in a significantly highest number of roots (Avg. 8) and root length (6.15 cm). Plantlets with healthy, multiple roots were transplanted in several soil combinations of river silt, bolhari (yellow sand), and peat moss. However, the significantly highest survival (100%) of plantlets in the greenhouse was obtained on the soil medium containing only peat moss. Furthermore, soil mixtures of bolhari and peat moss (1:1) and river silt alone exhibited 98.9% and 95.1% survivability of plantlets, which was also considered equally significant (at 5% probability level). The in vitro and ex vitro protocols optimized in the current study can be applied commercially for pineapple crop worldwide.

The excessive use of chemical fertilizers is deteriorating both the environment and soil, making it a big challenge faced by sustainable agriculture. To assist the efforts for the solution of this burning issue, nine different potential native strains of plant growth-promoting bacteria (PGPB) namely, SA-1(Bacillus subtilis), SA-5 (Stenotrophomonas humi),SA-7(Azospirillum brasilense), BH-1(Azospirillum oryzae), BH-7(Azotobacter armeniacus), BH-8(Rhizobium pusense), BA-3(Azospirillum zeae), BA-6(Rhizobium pusense), and BA-7(Pseudomonas fragi) were isolated that were characterized morphologically, biochemically and molecularly on the basis of 16S rRNA sequencing. Furthermore, the capability of indigenous PGPB in wheat (Triticum aestivum, Chakwal-50) under control, DAP+FYM, SA-1,5,7, BH-1,7,8, BA-3,6,7, DAP+ FYM + SA-1,5,7, DAP+FYM+ BH-1,7,8 and DAP+FYM+ BA-3,6,7 treatments was assessed in a randomized complete block design (RCBD). The results of the study showed that there was a significant increase in plant growth, nutrients, quality parameters, crop yield, and soil nutrients at three depths under SA-1,5,7, BH-1,7,8, and BA-3,6,7 in combination with DAP+FYM. Out of all these treatments, DAP+ FYM + BA-3,6,7 was found to be the most efficient for wheat growth having the highest 1000-grain weight of 55.1 g. The highest values for plant height, no. of grains/spike, spike length, shoot length, root length, shoot dry weight, root dry weight, 1000 grain weight, biological yield, and economic yield were found to be 90.7 cm, 87.7 cm, 7.20 cm, 53.5 cm, 33.5 cm, 4.87 g, 1.32 g, 55.1 g, 8209 kg/h, and 4572 kg/h, respectively, in the DAP+FYM+BA treatment. The DAP+FYM+BA treatment had the highest values of TN (1.68 µg/mL), P (0.38%), and K (1.33%). Likewise, the value of mean protein (10.5%), carbohydrate (75%), lipid (2.5%), and available P (4.68 ppm) was also highest in the DAP+FYM+BA combination. C:P was found to be significantly highest (20.7) in BA alone but was significantly lowest (11.9) in DAP+FYM+BA. Hence, the integration of strains BA-3, BA-5, and BA-7 in fertilizers can be regarded as the most suitable choice for agricultural growth in the sub-mountainous lower region of AJK. This could serve as the best choice for sustainable wheat growth and improved soil fertility with lesser impacts on the environment.

The glutamate-gated ion channels known as N-methyl-d-aspartate receptors (NMDARs) are important for both normal and pathological brain function. Subunit-selective antagonists have high therapeutic promise since many pathological conditions involve NMDAR over activation, although few clinical successes have been reported. Allosteric inhibitors of GluN2B-containing receptors are among the most potential NMDAR targeting drugs. Since the discovery of ifenprodil, a variety of GluN2B-selective compounds have been discovered, each with remarkably unique structural motifs. These results expand the allosteric and pharmacolog-ical spectrum of NMDARs and provide a new structural basis for the development of next-generation GluN2B antagonists that have therapeutic potential in brain diseases. Small molecule therapeutic inhibitors targeting NMDA have recently been developed to target CNS disorders such as Alzheimer’s disease. In the current study, a cheminformatics method was used to discover potential antagonists and to identify the structural requirements for Gly/NMDA antagonism. In this case we have created a useful pharmacophore model with solid statistical values. Through pharmacophore mapping, the verified model was used to filter out virtual matches from the ZINC database. Assessing receptor-ligand binding mechanisms and affinities used molecular docking. To find the best hits, the GlideScore and the interaction of molecules with important amino acids were considered essential features. We found some molecular inhibitors, namely, ZINC13729211, ZINC07430424, ZINC08614951, ZINC60927204, ZINC12447511, and ZINC18889258 with high binding affinity using computational methods. The molecules in our studies showed characteristics such as good stability, hydrogen bonding and higher binding affinities in the solvation-based assessment method than ifenprodil with acceptable ADMET profile. Moreover, these six leads have been proposed as potential new perspectives for exploring potent Gly/NMDA receptor antagonists. In addition, it can be tested in the laboratory for potential therapeutic strategies for both in vitro and in vivo research.

Portulacaria afra Jacq (Portulacaceae) is a medicinal plant used traditionally in the treatment of pain and inflammation. This study aimed to evaluate the phytochemical composition, toxicity, antinociceptive, and enzyme inhibition potential of P. afra. The methanol extract (PAME) was prepared through maceration and fractionated with ethanol to ethanol fraction of P. afra (ETPA). Both PAME and ETPA were found to be rich in total phenolic (TPC) and total flavonoid contents (TFC). Similarly, PAME showed the highest antioxidant potential through ABTS 93.16 ± 0.05 mg TE /g of dry extract (D.E.) while ETPA showed maximum results (462 ± 1.44 mg TE/g) in the CUPRAC method. The RP-UHPLC-MS analysis of PAME showed tentative identification of 101 compounds in the positive mode and 14 compounds in the negative mode of ionization while GC–MS profiling gave a total of 15 compounds. The acute oral toxicity study of PAME revealed the safety and biocompatibility of the extract up to a dose of 5000 mg/kg orally in rats. In-vitro data revealed that varying concentrations of P. afra significantly (p

Organic acids (3-chlorobenzoic acid and 4-chlorobenzoic acid) were treated with 4-amino-2-chloropyridine and 2-amino-4-chloropyridine as coformers for cocrystallization. Acid/base pairs afforded a cocrystal (3-chlorobenzoic acid and 4-amino-2-chloropyridine, compound 1) and molecular salts (2-amino-4-chloropyridinium 3-chlorobenzoate, 2; 2-amino-4-chloropyridinium 4-chlorobenzoate, 3). The products were characterized with the help of FT-IR, UV/visible spectroscopy, PXRD, and SC-XRD. The position of the Cl-substituent on the phenyl ring was explored with respect to proton transfer between acid/base pairs, and its role in supramolecular chemistry is discussed in detail. The existence of supramolecular assemblies was further reproduced using DFT calculations. Further, frontier molecular orbital (FMO), molecular electrostatic potential (MEP), and noncovalent interaction index (NCI) analyses were performed to gain insight into the electronic properties and nature of noncovalent interactions. The prepared compounds were examined for their biological activities against selected Gram-positive and Gram-negative bacterial strains and were also tested as antioxidant agents (DPPH free radical scavenging). Structural parameters determined experimentally and theoretically are within the expected range, and the biological activities of the resultant compounds are moderate.

Sesuvium sesuvioides (Fenzl) Verdc (Aizoaceae) has been traditionally used in the treatment of inflammation, arthritis, and gout. However, its antiarthritic potential has not been evaluated scientifically. The current study was designed to assess the antiarthritic properties of the n-butanol fraction of S. sesuvioides (SsBu) by phytochemical analysis, in vitro and in vivo pharmacological activities, and in silico studies. Phytochemical analysis showed total phenolic contents (90.7 ± 3.02 mg GAE/g) and total flavonoid contents (23.7 ± 0.69 mg RE/g), and further analysis by GC-MS identified possible bioactive phytocompounds belonging to phenols, flavonoids, steroids, and fatty acids. The in vitro antioxidant potential of SsBu was assessed by DPPH (175.5 ± 7.35 mg TE/g), ABTS (391.6 ± 17.1 mg TE/g), FRAP (418.2 ± 10.8 mg TE/g), CUPRAC (884.8 ± 7.97 mg TE/g), phosphomolybdenum (5.7 ± 0.33 mmol TE/g), and metal chelating activity (9.04 ± 0.58 mg EDTAE/g). Moreover, in the in vitro studies, inhibition (%) of egg albumin and bovine serum albumin denaturation assays showed that the anti-inflammatory effect of SsBu at the dose of 800 μg/ml was comparable to that of diclofenac sodium used as a standard drug. The in vivo antiarthritic activity was assessed to determine the curative impact of SsBu against formalin-induced (dose-dependent significant (p < 0.05) effect 72.2% inhibition at 750 mg/kg compared to standard; 69.1% inhibition) and complete Freund’s adjuvant-induced arthritis (40.8%; inhibition compared to standard, 42.3%). SsBu significantly controlled PGE-2 level compared to the control group (p < 0.001) and restored the hematological parameters in rheumatoid arthritis. Treatment with SsBu significantly reduced oxidative stress by reinstating superoxide dismutase, GSH, and malondialdehyde along with pro-inflammatory markers (IL-6 and TNF-α) in arthritic rats. Molecular docking revealed the antiarthritic role of major identified compounds. Kaempferol-3-rutinoside was found to be more potent for COX-1 (−9.2 kcal/mol) and COX-2 inhibition (−9.9 kcal/mol) than diclofenac sodium (COX-1, −8.0 and COX-2, −6.5 kcal/mol). Out of the 12 docked compounds, two for COX-1 and seven for COX-2 inhibition showed more potent binding than the standard drug. The results from the in vitro, in vivo, and in silico approaches finally concluded that the n-butanol fraction of S. sesuvioides had antioxidant and antiarthritic potential, which may be due to the presence of potential bioactive compounds.

This paper contains the comparative study of elasto-mechanical, electronic and optical properties of pyrochlores. These structures are first optimized and second order-independent cubic elastic constants C11, C12 and C44 are computed using the optimized data. These computed cubic elastic constants are further used to check the mechanical stability, Poisson's ratio, Pugh's ratio and Cauchy pressure which depicts the flexible and ductile nature of both materials. Kleinman ratio is more significant for La2Zr2O7, so it has excellent bond stretching minimization phenomena. Young's modulus and lame's constants describe that Nd2Zr2O7 is stiffer and difficult to compress than La2Zr2O7. Debye temperature and sound velocities are also calculated for both oxides. Due to the high melting temperature, Nd2Zr2O7 is suitable for high-temperature environments and is used in aerospace and construction industries. Band structures reveal that these oxides are metallic as there appears no gap in electronic dispersion relations. Analysis of the density of states reveals the magnetic nature of these oxides as the density of states is not the perfect image of each other in both spin channels. The imaginary part of the dielectric function also shows the optically metallic nature and suitability for sensors and opto-electronic devices.

Abstract Terpenoids from natural plant sources are valuable for their diverse biological activities that have important roles in the medical and agrochemical industries. In this study, we assessed the antioxidant, antifungal, and aphicidal activities of a mixture of spinasterol and 22,23-dihydrospinasterol from the leaves of Citrullus colocynthis. We used 1,1-diphenyl-2-picrylhydrazyl (DPPH) to assess antioxidant activity, and we measured antifungal activity using mycelium growth inhibition assays with three pathogenic fungi, Magnaporthe grisea, Rhizoctonia solani, and Phytophthora infestans. Aphicidal activity against adults of Myzus persicae was determined using in vitro and in vivo assays. Spinasterol and 22,23-dihydrospinasterol exhibited moderate antioxidant activity, even at lower concentrations: 19.98% at 0.78 µg mL−1, 31.52% at 3.0 µg mL−1, 36.61% at 12.5 µg mL−1, and 49.76% at 50 µg mL−1. Spinasterol and 22,23-dihydrospinasterol showed reasonable levels of fungicidal activity toward R. solani and M. grisea, with EC50 values of 129.5 and 206.1 µg mL−1 , respectively. The positive controls boscalid and carbendazim were highly effective against all fungi except boscalid for M. grisea (EC50 = 868 µg mL−1) and carbendazim for P. infestans (EC50 = 8721 µg mL−1). Significant insecticidal activity was observed in both residual and greenhouse assays, with LC50 values of 42.46, 54.86, and 180.9 µg mL−1 and 32.71, 42.46, and 173.8 µg mL−1 at 72, 48, and 24 h, respectively. The antioxidant activity of spinasterol and 22,23-dihydrospinasterol was strongly positively correlated with their antifungal and insecticidal activity. Spinasterol and 22,23-dihydrospinasterol therefore show good antioxidant and aphicidal activity with moderate fungicidal activity, making them suitable candidates for an alternative to synthetic agents.

The main objective of this work was to gain insight into biological propensities, and bioactive phytochemicals of Acacia jacquemontii Benth, a wild plant providing medicinal components, as well as to establish a link between its phytochemical profile and biological activities. Phytochemical profiling revealed the presence of a higher amount of total phenolic (271.44 ± 4.41 mg GAE/g) and flavonoid contents (216.47 ± 5.82 mg QE/g) in methanolic extract (MEAJ), and as compared to n-hexane fraction (HEAJ) and stronger biological activities of MEAJ were possibly linked to the higher bioactive contents. The freshly collected plant leaves showed a strong antioxidant potential (total antioxidant capacity 1.03 ± 0.19 mmol TE/g), which was found even stronger in dried methanolic extract (TAC; 4.36 ± 1.12 mmol TE/g), moreover, MEAJ also showed strong antioxidant potential when investigated by different antioxidant assays (DPPH; 154.04 ± 2.47, ABTS; 122.36 ± 0.80, FRAP; 453.18 ± 5.9, CUPRAC; 1389.97 ± 5.32 mg TE/g). The MEAJ showed good tyrosinase inhibition activity (71.69 %), compared with 83 % inhibition by kojic acid. Ten major compounds identified by GC–MS were docked and eight legends showed lower binding energies (-6 to −7.8 kcal/mol) compared with kojic acid (-5.9 kcal/mol), which shows the possible role of these compounds in the anti-tyrosinase activity of the extract, and the ADMET analysis predicted the drug-likeness and safety profile of the studied compounds. The thrombolytic effect of MEAJ was 56.41 ± 0.75 to 57.15 ± 1.41 % which was comparable with streptokinase (82.44 ± 1.15 to 84.14 ± 0.95 %). Antibacterial activity of MEAJ was also good (MEAJ; 0.5–2.0 mg/mL, and co-amoxiclav; 5.0–12.5 µg/mL), and the highest activity was observed against Bacillus subtilis (MEAJ; 0.5 mg/mL, co-amoxiclav; 5.0 µg/mL). The antiviral activity of MEAJ was highly strong (HA titer; 00 to 08) against all the tested strains. It can be concluded that A. jacquemontii is a prospective source of phytochemicals with strong biological activities, and their usage in formulations of natural products and pharmaceuticals is recommended, however, further research may address the discovery and development of novel drugs for the pharmaceutical industry.

A comparative study between methanolic extract and n-hexane fraction of Typha domingensis (Typhaceae) was conducted for the evaluation of phytochemical potential, in vitro biological activities, and in-silico molecular docking studies. The phytochemical composition was estimated by total phenolic and total flavonoid contents, and by GC–MS analysis. Several biological activities were performed such as antioxidant assays (ABTS, FRAP, DPPH, & CUPRAC), enzyme inhibition activity (Tyrosinase, Acetylcholinesterase & Butyrylcholinesterase), thrombolytic activity, and antimicrobial activity (antibacterial & antiviral) to evaluate the medicinal importance of Typha domingensis. The results of the comparative study showed that methanolic extract has more total phenolic and total flavonoid contents (95.72 ± 5.76 mg GAE/g, 131.66 ± 7.92 mg QE/g, respectively) as compared to n-hexane fraction which confirms its maximum antioxidant potential (ABTS 114.31 ± 8.17, FRAP 116.84 ± 3.01, DPPH 283.54 ± 7.3 & CUPRAC 284.16 ± 6.5 mg TE/g). In the case of in vitro enzyme inhibition study and thrombolytic activity, better results were observed for methanolic extract. Almost similar antimicrobial patterns were observed for both methanolic extract and n-hexane fraction of Typha domingensis. The major bioactive phytochemicals identified by GC–MS were further analyzed for in-silico molecular docking studies to determine the binding affinity between ligands and the enzymes. The docking study indicated that most of the bioactive compounds showed a better binding affinity with enzymes as compared to the standard compounds (kojic acid & galantamine). The results of this study recommended that Typha domingensis has promising pharmaceutical importance and it should be further analyzed for the isolation of bioactive phytochemicals which may be useful for the treatment of several diseases.

The widespread impacts of projected global and regional climate change on rice yield have been investigated by different indirect approaches utilizing various simulation models. However, direct approaches to assess the impacts of climatic variabilities on rice growth and development may provide more reliable evidence to evaluate the effects of climate change on rice productivity. Climate change has substantially impacted rice production in the mid-high latitudes of China, especially in Northeast China (NEC). Climatic variabilities occurring in NEC since the 1970s have resulted in an obvious warming trend, which made this region one of the three major rice-growing regions in China. However, the projections of future climate change have indicated the likelihood of more abrupt and irregular climatic changes, posing threats to rice sustainability in this region. Hence, understanding the self-adaptability and identifying adjustive measures to climate variability in high latitudes has practical significance for establishing a sustainable rice system to sustain future food security in China. A well-managed field study under randomized complete block design (RCBD) was conducted in 2017 and 2018 at two study sites in Harbin and Qiqihar, located in Heilongjiang province in NEC. Four different cultivars were evaluated: Longdao-18, Longdao-21 (longer growth duration), Longjing-21, and Suijing-18 (shorter growth duration) to assess the inter-relationships among grain-filling parameters, grain yield and yield components, and grain quality attributes. To better compare the adaptability mechanisms between grain-filling and yield components, the filling phase was divided into three sub-phases (start, middle, and late). The current study evaluated the formation and accumulation of the assimilates in superior and inferior grains during grain-filling, mainly in the middle sub-phase, which accounted for 59.60% of the yield. The grain yields for Suijing-18, Longjing-21, Longdao-21, and Longdao-18 were 8.02%, 12.78%, 17.19%, and 20.53% higher in Harbin than those in Qiqihar, respectively in 2017, with a similar trend observed in 2018. At Harbin, a higher number of productive tillers was noticed in Suijing-18, with averages of 17 and 15 in 2017 and 2018, respectively. The grain-filling parameters of yield analysis showed that the filling duration in Harbin was conducive to increased yield but the low dry weight of inferior grains was a main factor limiting the yield in Qiqihar. The average protein content values in Harbin were significantly higher (8.54% and 9.13%) than those in Qiqihar (8.34% and 9.14%) in 2017 and 2018, respectively. The amylose content was significantly higher in Harbin (20.03% and 22.27%) than those in Qiqihar (14.44% and 14.67%) in 2017 and 2018, respectively. The chalkiness percentage was higher in Qiqihar, indicating that Harbin produced good quality rice. This study provides more direct evidence of the relative changes in rice grain yield due to changes in grain-filling associated with relative changes in environmental components. These self-adaptability mechanisms to climatic variability and the inter-relationships between grain-filling and grain yield underscore the urgent to investigate and explore measures to improve Japonica rice sustainability, with better adaptation to increasing climatic variabilities. These findings may also be a reference for other global rice regions at high latitudes in addressing the impacts of climate change on future rice sustainability.

Yaks are often subject to long-term starvation and a high prevalence of respiratory diseases and mortality in the withered season, yet the mechanisms that cause this remain unclear. Research has demonstrated that β-hydroxybutyrate (BHB) plays a significant role in regulating the immune system. Hence, we hypothesize that the low glucose and high BHB condition induced by severe starvation might have an effect on the pro-inflammatory response of the alveolar macrophages (AMs) in yaks. To validate our hypothesis, we isolated and identified primary AMs from freshly slaughtered yaks and cultured them in a medium with 5.5 mM of glucose or 2.8 mM of glucose plus 1–4 mM of BHB. Utilizing a real-time quantitative polymerase chain reaction (RT-qPCR), immunoblot assay, and enzyme-linked immunosorbent assay (ELISA), we evaluated the gene and protein expression levels of GPR109A (G-protein-coupled receptor 109A), NF-κB p65, p38, and PPARγ and the concentrations of pro-inflammatory cytokines interleukin (IL)-1β and IL-6 and tumor necrosis factor (TNF)-α in the supernatant. The results demonstrated that AMs exposed to low glucose plus BHB had significantly higher levels of IL-1β, IL-6, and TNF-α (p < 0.05) and higher activity of the GPR109A/NF-κB signaling pathway. A pretreatment of either pertussis toxin (PTX, inhibitor of GPR109A) or pyrrolidinedithiocarbamic (PDTC, inhibitor of NF-κB p65) was effective in preventing the elevated secretion of pro-inflammatory cytokines induced by low glucose plus BHB (p < 0.05). These results indicated that the low glucose plus BHB condition would induce an enhanced pro-inflammatory response through the activation of the GPR109A/NF-κB signaling pathway in primary yak AMs, which is probably the reason why yaks experience a higher rate of respiratory diseases and mortality. This study will offer new insight into the prevention and treatment of bovine respiratory diseases.

Glomerella leaf spot (GLS) is a severe infectious disease of apple whose infective area is growing gradually and thus poses a huge economic threat to the world. Different species of Colletotrichum including Colletotrichum gloeosporioides are responsible for GLS. For efficient GLS control, it is important to understand the mechanism by which the cruciferous crops and C. gloeosporioides interact. Arginine is among one of the several types of amino acids, which plays crucial role in biochemical and physiological functions of fungi. The arginine biosynthesis pathway involved in virulence among plant pathogenic fungi is poorly understood. In this study, CgCPS1 gene encoding carbamoyl phosphate synthase involved in arginine biosynthesis has been identified and inactivated experimentally. To assess the effects of CgCPS1, we knocked out CgCPS1 in C. gloeosporioides and evaluated its effects on virulence and stress tolerance. The results showed that deletion of CgCPS1 resulted in loss of pathogenicity. The Δcgcps1 mutants showed slow growth rate, defects in appressorium formation and failed to develop lesions on apple leaves and fruits leading to loss of virulence while complementation strain (CgCPS1-C) fully restored its pathogenicity. Furthermore, mutant strains showed extreme sensitivity to high osmotic stress displaying that CgCPS1 plays a vital role in stress response. These findings suggest that CgCPS1 is major factor that mediates pathogenicity in C. gloeosporioides by encoding carbamoyl phosphate that is involved in arginine biosynthesis and conferring virulence in C. gloeosporioides.

Abstract Bemisia tabaci is one of the most notorious agricultural pests in the world. A vicious circle among insect resistance, dose increased, environment and human body impaired as the overuse of synthetic pesticides are becoming increasingly evident. Entomopathogenic Beauveria sp. is known as an effective natural enemy to control B. tabaci. Therefore, this study aimed to purify and identify the biological compounds from Beauveria sp. LY2 via extensive chromatographic techniques, NMR and MS and evaluated for their insecticidal activities against B. tabaci via contact and feeding assay. The outcome identified that one new cerebroside, cerebroside F (1), nine known compounds, cerebroside B (2), bassiatin (3), methyl 1,4-dihydro-4-oxo-2-quinolinecarboxylate (4), cerevisterol (5), 9-hydroxycerevisterol (6), 6-dehydrocerevisterol (7), (22E,24R)-ergosta-8(14),22-diene-3β,5α,6β,7α-tetrol (8), melithasterol B (9) and ergosterol peroxide (10) were isolated. Among the known compounds, methyl 1,4-dihydro-4-oxo- 2-quinolinecarboxylate (4) was isolated from natural origin for the first time. It is demonstrable from the results that compounds 3, 4 and 7 strongly featured insecticidal activities against B. tabaci, being the LC50 value as 10.59, 19.05, 26.59 μg/mL respectively in contact as well as 11.42, 5.66, 5.65 μg/mL respectively in feeding experiment. Moreover, no adverse effect on plant growth/height or phytotoxicity was observed on pepper, cucumber, tomato and cotton. The data from the current study has provided the foundation for the use of newly purified compounds against Bemisia tabaci as an alternative to synthetic chemical compounds.

Objectives: Aloe perryi plant in the genus Aloe (family: Asphodelaceae), have been used in traditional medicine systems. Herein, we aimed to analyze the anticancer effects of A. perryi petroleum ether extract (APPeE) against human breast cancer (MDA-MB-231) and normal (HEK-293) cell lines. Further the biomarker stigmasterol, isolated from A. perryi extract was quantified by a densitometric high-performance thin layer chromatography (HPTLC) method. Methods: The cytotoxic potential of APPeE was measured by MTT assay, neutral red uptake (NRU) assay, and morphological identification. Results: APPeE-induced a concentration dependent strong cytotoxic effects on MDA-MB-231 cells with an IC50 value of 24.5 μg/ml in contrast to HEK-293 (IC50 value of > 100 μg/ml). Similar results were obtained by NRU assay. Further cytotoxic concentrations of APPeE increased reactive oxygen species (ROS) production and activated caspase-3 and −9, leading to apoptosis in MDA-MB-231 cells. Additionally, a significant augment in the expression of p53, Bax, caspase-3 and −9 with a decline in Bcl-2 gene expression confirmed the involvement of apoptosis pathway in MDA-MB-231 cell death. In HPTLC analysis, the solvent system hexane: ethylacetate (8:2 v/v) furnished a sharp peak for stigmasterol (Rf = 0.19 ± 0.001). The low value of % relative standard deviation (RSD) (0.97–1.39) proposed the method is robust. The limit of detection (LOD) and limit of quantification (LOQ) were established as 13.82 and 41.89 ng, respectively. The stigmasterol in APPeE was quantified as 0.238% w/w of dried APPeE. Conclusions: Hence, this study concludes the promising anticancer effect of APPeE, which might be endorsed to the existence of known anticancer biomarker stigmasterol.

Population explosion, industrialization, and urbanization have accelerated energy requirements across the globe. This has led to the human quest to find simple and cost-effective energy solutions. A promising solution is the revival of the Stirling engine with the addition of Shape Memory Alloy NiTiNOL in it. The experimental results reveal that the addition of a NiTiNOL spring at the base plate of the Stirling engine enhances the overall efficiency of the engine, demonstrating some impact of the shape memory alloy toward the performance output of the Stirling engine. The newly modified engine has been named the STIRNOL ENGINE. The comparative study of Stirling and Stirnol engines reveals a minuscule efficiency improvement, yet there is a furtherance that opens a window for future researchers to get a lead and venture into this new field. We are confident that with more complex designs and better Stirling and NiTiNOL combinations, more efficient engines can be invented in the future. This research focuses on changing the material of the base plate of the Stirnol engine and ascertaining its performance differential through the integration of the NiTiNOL spring. A minimum of four types of materials are utilized for experimentation.

Neurological disorders are disorders characterized by progressive loss of neurons leading to disability. Neurotransmitters such as nor-adrenaline, dopamine, and serotonin are partially regulated by the enzyme monoamine oxidase (MAO). Treatments for conditions like Alzheimer’s, Parkinson’s, anxiety, and depression involve the use of MAOIs. To target MAO enzyme inhibition, various scaffolds are prepared and evaluated, including modified coumarins, chromone carboxylic acid substituents, pyridazine derivatives, and indolylmethylamine. The research presented here focuses on combining different computational tools to find new inhibitors of the MAO-B protein. We discovered 5 possible chemical inhibitors using the above computational techniques. We found five molecular inhibitors with high binding affinity using computational methods. These five molecules showed a high binding affinity; they are −10.917, −10.154, −10.223, −10.858, and −9.629 Kcal/mol, respectively. Additionally, the selected inhibitors were further examined by in vitro activity, and their binding affinity was confirmed using an enzyme-based assay. In summary, the computational studies performed here using molecular dynamics and free energy calculations can also be used to design and predict highly potent derivatives as MAO-B inhibitors, and these top inhibitors help in the development of novel drugs for neurological diseases such as Alzheimer’s and Parkinson’s.

Rice blast is a major threat to the rice crop in rice-growing areas around the globe. However, synthetic fungicides are injurious for the ecosystem and the existing bio-control agents facing the resistance from the pathogens. In this study, an isolated novel Streptomyces strain (TA-47), from the rice field was identified by culturing on different media, physiological tests, biochemical tests, chemo-taxonomy and molecular biological approaches. In-vitro and in-vivo bio-control activity of the isolated strain was evaluated. Growth on the media, spore production, utilization of sole carbon and nitrogen sources, presence of diaminopimelic acid (DAP) in the cell wall of the strain and fatty acid production were observed.16S rRNA (gene sequence) and a neighbour-joining hood tree indicated that the strain TA-47 is Streptomyces koyanogensis. In-vitro inhibition zone of 29 mm against the M. grisea showed the antagonistic potential of the novel strain TA-47. On the other hand, in the in-vivo studies, strain TA-47, showed the strong bio-control effect, as disease severity decreased to 19.66 ± 1.6%, while in the non-treated sample the disease severity was recorded as 44.66 ± 5.249%. Moreover, after the treatment of the strain, effects on the growth, such as increased height and root length by 72.22 and 16.54 cm, respectively, were also recorded. In conclusion, rhizosphere Streptomyces TA-47, was a novel Streptomyces koyanogensis strain. The study found that the strain had strong bio-control and growth-promoting abilities. Abbreviation: M Grisea, Magnaporthe grisea; ISP, International Streptomyces Project; PDA, Potato Dextrose AgarSEM, Scan Electron Microscope; TLC, Thin Layer Chromatography; GC-MS, Gas Chromatography Mass Spectrum

Objective: To explore the factors which motivate or hamper our youngsters for blood donation and to understand the behavior of youth regarding blood donation through which we can rise voluntary blood donation. Study Design: Cross sectional study. Place and Duration of Study: Allama Iqbal Medical College Lahore, from Jan 2017 to Sep 2017. Methodology: A self-designed detailed questionnaire was designed, comprised of the demographic status of the subject, various aspects of the attitudes, perceptions, and drives toward blood donation including the questions regarding earlier blood donation, their experience, future planning for blood donation and reasons for not donating the blood. Results: Out of the 789 participants, only 284 (36.01%) were blood donors including 231 (81.39%) males and 53 18.61%) females. Altruism, sense of social responsibility, volunteerism, spiritual pleasure and gaining experience were major factors for blood donation while never asked for donation, objection from elders, fear for the needle, fear of adverse effects, weight gain/loss were the barriers which impede our youngsters for blood donation. Conclusion: Blood donation can be up surged with proper counseling, diminishing out spread myths and false perception among young population of Pakistan.

The hormonal imbalances, including abscisic acid (ABA) and brassinosteroid (BR) levels, caused by salinity constitute a key factor in hindering spikelet development in rice and in reducing rice yield. However, the effects of ABA and BRs on spikelet development in plants subjected to salinity stress have been explored to only a limited extent. In this research, the effect of ABA and BRs on rice growth characteristics and the development of spikelets under different salinity levels were investigated. The rice seedlings were subjected to three different salt stress levels: 0.0875 dS m−1 (Control, CK), low salt stress (1.878 dS m−1, LS), and heavy salt stress (4.09 dS m−1, HS). Additionally, independent (ABA or BR) and combined (ABA+BR) exogenous treatments of ABA (at 0 and 25 μM concentration) and BR (at 0 and 5 μM concentration) onto the rice seedlings were performed. The results showed that the exogenous application of ABA, BRs, and ABA+BRs triggered changes in physiological and agronomic characteristics, including photosynthesis rate (Pn), SPAD value, pollen viability, 1000-grain weight (g), and rice grain yield per plant. In addition, spikelet sterility under different salt stress levels (CK, LS, and HS) was decreased significantly through the use of both the single phytohormone and the cocktail, as compared to the controls. The outcome of this study reveals new insights about rice spikelet development in plants subjected to salt stress and the effects on this of ABA and BR. Additionally, it provides information on the use of plant hormones to improve rice yield under salt stress and on the enhancement of effective utilization of salt-affected soils.

The development of efficient cell culture strategies for the generation of dopaminergic neurons is an important goal for transplantation-based approaches to treat Parkinson’s disease. To identify extracellular matrix molecules that enhance differentiation and might be used in these cell cultures we have used micro-contact printed arrays on glass slides presenting 190 combinations of 19 extracellular matrix molecules selected on the basis of their expression during embryonic development of the ventral midbrain. Using long-term neuroepithelial stem cells (Lt-NES), this approach identified a number of matricellular proteins that enhanced differentiation, with the combination of Sparc, Sparc-like (Sparc-l1) and Nell2 increasing the number of tyrosine hydroxylase+ neurons derived from Lt-NES cells and, critically for further translation, human pluripotent stem cells.

The current study was designed to investigate the feasibility of incorporating the water-insoluble lipophilic drug Alprazolam (Alp) into solid lipid nanoparticles (SLNs) to offer the combined benefits of the quick onset of action along with the sustained release of the drug. Therefore, compritol-based alprazolam-loaded SLNs (Alp-SLNs) would provide early relief from anxiety and sleep disturbances and long-lasting control of symptoms in patients with depression, thereby enhancing patient compliance. The optimized Alp-SLNs analyzed by DLS and SEM showed consistent particle size of 92.9 nm with PI values and standard deviation of the measurements calculated at

Air quality has a significant influence on the environment and health. Instruments that efficiently and inexpensively detect air quality could be extremely valuable in detecting air quality indices. This study presents a robust deep learning model named AQE-Net, for estimating air quality from mobile images. The algorithm extracts features and patterns from scene photographs collected by the camera device and then classifies the images according to air quality index (AQI) levels. Additionally, an air quality dataset (KARACHI-AQI) of high-quality outdoor images was constructed to enable the model’s training and assessment of performance. The sample data were collected from an air quality monitoring station in Karachi City, Pakistan, comprising 1001 hourly datasets, including photographs, PM2.5 levels, and the AQI. This study compares and examines traditional machine learning algorithms, e.g., a support vector machine (SVM), and deep learning models, such as VGG16, InceptionV3, and AQE-Net on the KHI-AQI dataset. The experimental findings demonstrate that, compared to other models, AQE-Net achieved more accurate categorization findings for air quality. AQE-Net achieved 70.1% accuracy, while SVM, VGG16, and InceptionV3 achieved 56.2% and 59.2% accuracy, respectively. In addition, MSE, MAE, and MAPE values were calculated for our model (1.278, 0.542, 0.310), which indicates the remarkable efficacy of our approach. The suggested method shows promise as a fast and accurate way to estimate and classify pollutants from only captured photographs. This flexible and scalable method of assessment has the potential to fill in significant gaps in the air quality data gathered from costly devices around the world.

The purpose of this study was to find the biological propensities of the vegetable plant Pleurospermum candollei by investigating its phytochemical profile and biological activities. Phytochemical analysis was done by spectroscopic methods to investigate the amount of total polyphenols, and biological evaluation was done by the different antioxidant, enzyme inhibitory (tyrosinase, α-amylase, and α-glucosidase), thrombolytic, and antibacterial activities. The highest amount of total phenolic and flavonoid contents was observed in methanolic extract (240.69 ± 2.94 mg GAE/g and 167.59 ± 3.47 mg QE/g); the fractions showed comparatively less quantity (57.02 ± 1.31 to 144.02 ± 2.11 mg GAE/g, and 48.21 ± 0.75 to 96.58 ± 2.30 mg QE/g). The effect of these bioactive contents was also related to biological activities. GCMS analysis led to the identification of bioactive compounds with different biological effects from methanolic extract (antioxidant; 55.07%, antimicrobial; 56.41%), while the identified compounds from the n-hexane fraction with antioxidant properties constituted 67.86%, and those with antimicrobial effects constituted 82.95%; however, the synergetic effect of polyphenols may also have contributed to the highest value of biological activities of methanolic extract. Molecular docking was also performed to understand the relationship of identified secondary metabolites with enzyme-inhibitory activities. The thrombolytic activity was also significant (40.18 ± 1.80 to 57.15 ± 1.10 % clot lysis) in comparison with streptokinase (78.5 ± 1.53 to 82.34 ± 1.25% clot lysis). Methanolic extract also showed good activity against Gram-positive strains of bacteria, and the highest activity was observed against Bacillus subtilis. The findings of this study will improve our knowledge of phytochemistry, and biological activities of P. candollei, which seems to be a ray of hope to design formulations of natural products for the improvement of health and prevention of chronic diseases; however, further research may address the development of novel drugs for use in pharmaceuticals.

The current study describes ethnobotanical (EB) and traditional ethnomedicinal (TEMs) uses of medicinal plants (MPs) of Tehsil Samahni area of Azad Jammu and Kashmir, Pakistan. Indigenous inhabitants of the area mostly reside in remote mountainous areas and depend on wild plants for life necessities and treatment of different diseases. This paper presents first quantitative ethnobotanical study describing the popularity level of MPs in indigenous communities by using relative popularity level (RPL) and rank order of priority (ROP) indices. The data of TEMs and EB uses were collected from 200 interviewees consisting of traditional herbal practioners (110) and farmers (90) from study area using structured and semi-structured interview methodology during year 2019. It was found that illiterate peasants have more knowledge and practice of TEMs than other people. A total of 150 plant species belonging to 58 families with botanical and local names, parts used, mode of recipes preparation, administration mechanism and TEMs uses were documented in systematic manner. It was explored that highest number (7.69%) of plants of Moraceae were used in TEMs and followed by species of Asteraceae (6.29%). The ethnobotanical data was analyzed by using quantitative ethnobotanical tools viz: informant consensus factor (ICF), fidelity level (FL), relative frequency of citation (RFC), use value index (UVI), relative importance of plants (RIP), relative populatiry level (RPL) and rank order of popularity (ROP). The leaf ranked first (36.49%) and fruit 2nd (12.07%) being used in TEMs while prevalent use-form was decoction (29.28%), followed by tea (23.05%) and preferably taken as oral (46.66%). ICF analysis proved that diabetes, hypertension, rabies, snake sting and joint pains were the most prevalent occurring infirmities. The highest RFC (0.95) was found for Acacia modesta, followed by Boerhavia procumbens (RFC:0.87) and Berberis lycium (RFC:0.85). The relative importance of MPs was calculated by using UVI and Moringa oleifera showed highest (1.38), followed by Zanthoxylum armatum (1.25) and Withania somnifera (1.24) use-values. High UVI of plants depicts that these species are predominantly used in local health care system. The plants Phyllanthus emblica, Morus macruora, Justicia adhatoda and Melia azedarach depicted high values (1.00) of FL and RPL proving that these taxa are commonly used in indigenous medicines by local inhabitants and have better potential for drug discovery by ethnopharmacological analysis. Out of total, 26 species had more than 50% ROP index and Phyllanthus emblica and Flacourtia indica (ROP = 100) followed by Morus macruora (ROP: 97) were used to cure 'hypertension and hyperlipidemia' and 'cough, chest infection', respectively. It was found that 30% of total species were previously reported as medicinal while 70% plants were first time reported to be used in TEMs and authenticated by using of quantitative ethnobotanical tools. Ethnopharmacological potential of indigenous plants was confirmed by RIP and RPL indices which had been used to cure one or more body systems and were promulgated in the local herbal medicine system. The research provides clues to screen these plant species by using latest phytochemical and pharmacological analysis for novel drug discovery. This study will also be useful for conservation of bioculture traditional knowledge of indigenous communities and the most important is to conserve medicinal plants of the study area for future generations.

Silybum marianum L. is a therapeutic plant belonging to the family Asteraceae, which has exhibited silymarin, a principal component used to cure various physiochemical disorders. The study appraised the phytochemical analysis, antioxidant activity and chemical analysis of an extract from the seed, stem and leaves. Qualitative and quantitative phytochemical analysis was evaluated by the Folin–Ciocalteu reagent method and aluminum chloride colorimetric method, respectively. While the antioxidant activity was determined by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and acetate buffer in ferric chloride (FRAP) assay, respectively, the chemical profile was evaluated by Gas Chromatography-Mass Spectrometry (GC-MS) assay. The study outcomes identified that alkaloids, glycosides, flavonoids, terpenoids, steroids and catcholic tannins were present in seed, stem and leaves extracts except for saponins and Gallic tannins. Whereas, phenols were absent only in seed extract. Quantitative analysis revealed the presence of phenols and flavonoids in appreciable amounts of 21.79 (GAE/g), 129.66 (QE/g) and 17.29 (GAE/g), 114.29 (QE/g) from the leaves and stem extract, respectively. Similarly, all extracts expressed reasonable DPPH inhibition (IC50) and FRAP reducing power such as 75.98, 72.39 and 63.21% and 46.60, 51.40 and 41.30 mmol/g from the seeds, stem and leaves extract, respectively. Additionally, chemical analysis revealed the existence of 6, 8 and 9 chemical compounds from the seeds, stem and leaves extract, respectively, corresponding to 99.95, 99.96 and 98.89% of the whole extract. The chemical compound, Dibutyl phthalate was reported from all extracts while, Hexadecanoic acid, methyl ester and Silane, (1,1-dimethylethyl), dimethyl (phenylmethoxy) were reported only from the seed and leaves extract. Moreover, Methyl stearate was also a major compound reported from all extracts except for seed extract. It is demonstrable that extracts from different parts of S. marianum possess significant antioxidant activity, as well as valuable chemical compounds accountable for therapeutic effects that might be incorporated as an alternative to synthetic chemical agents.

During the last few decades, worsening air quality has been diagnosed in many cities around the world. The accurately prediction of air pollutants, particularly, particulate matter 2.5 (PM2.5) is extremely important for environmental management. A Convolutional Neural Network (CNN) P-CNN model is presented in this paper, which uses seven different pollutant satellite images, such as Aerosol index (AER AI), Methane (CH4), Carbon monoxide (CO), Formaldehyde (HCHO), Nitrogen dioxide (NO2), Ozone (O3) and Sulfur dioxide (SO2), as auxiliary variables to estimate daily average PM2.5 concentrations. This study estimates daily average of PM2.5 concentrations in various cities of Pakistan (Islamabad, Lahore, Peshawar and Karachi) by using satellite images. The dataset contains a total of 2562 images from May-2019 to April-2020. We compare and analyze AlexNet, VGG16, ResNet50 and P-CNN model on every dataset. The accuracy of machine learning models was checked with Mean Absolute Error (MAE), Root Mean Square Error (RMSE) and Mean Absolute Percentage Error (MAPE). The results show that P-CNN is more accurate than other approaches in estimating PM2.5 concentrations from satellite images. This study presents robust model using satellite images, useful for estimating PM2.5 concentrations.

Summary: The development of midbrain dopaminergic (mDA) neurons is controlled by multiple morphogens and transcription factors. However, little is known about the role of extracellular matrix proteins in this process. Here we examined the function of roof plate-specific spondins (RSPO1-4) and the floor plate-specific, spondin 1 (SPON1). Only RSPO2 and SPON1 were expressed at high levels during mDA neurogenesis, and the receptor LGR5 was expressed by midbrain floor plate progenitors. Surprisingly, RSPO2, but not SPON1, specifically promoted the differentiation of mDA neuroblasts into mDA neurons in mouse primary cultures and embryonic stem cells (ESCs). In addition, RSPO2 was found to promote not only mDA differentiation, but also mDA neurogenesis in human ESCs. Our results thus uncover an unexpected function of the matricellular protein RSPO2 and suggest an application to improve mDA neurogenesis and differentiation in human stem cell preparations destined to cell replacement therapy or drug discovery for Parkinson disease. : Gyllborg and colleagues report on the function of the matricellular protein R-Spondin 2 (RSPO2) in dopaminergic neuron development. RSPO2 is dynamically expressed during midbrain development and promotes the dopaminergic differentiation of mouse and human neuroblasts. Furthermore, they show that RSPO2 induced dopaminergic neurogenesis in human stem cell cultures, suggesting a possible application of RSPO2 in cell replacement strategies for Parkinson disease. Keywords: extracellular matrix, RSPO2, progenitors, dopaminergic neurons, differentiation, neurogenesis, human embryonic stem cells, Wnt, cell replacement, Parkinson disease

Novel compositions of methylcellulose in ethylene, propylene and butylene glycol were investigated for their thermal gel formation. These compositions have previously been found useful for inkjet-printing-based additive manufacturing processes as support materials. Experimental techniques such as viscosity measurements between 20 °C–150 °C–20 °C, differential scanning calorimetry (DSC) and X-ray diffraction (XRD) were used and the results showed that the gel formation upon cooling is caused by polymer–polymer association. The results also show that, for methylcellulose, propylene glycol is a better solvent than ethylene glycol and butylene glycol. Since no chemical reaction is involved, these gels can be used as support materials for jetting-based additive manufacturing processes.

BackgroundIn Pakistan, many tuberculosis (TB) cases are not reported to the national surveillance system. An active case finding strategy in the form of conventional (simple) or innovative (infotainment) chest camps can contribute to diagnosing these missed cases.ObjectiveTo compare the yield in terms of TB patients detected at a simple chest camp (SCC) versus an infotainment chest camp (ICC) in rural areas.MethodsA cluster randomized controlled trial with 2 parallel arms was conducted in 4 districts of Pakistan from June 2012 to May 2013. Rural neighborhoods (n = 318) were randomly allocated in a ratio of 1:3 to receive either SCC or ICC. Incidence of TB (all forms and sputum smear positive [SS+]) and number needed to screen (NNS) to diagnose 1 TB case were calculated. Cluster analysis was done according to intention to treat and risk ratio (RR), and 95% confidence intervals (CIs) were calculated.FindingsA total of 3086 participants were tested at the SCC and 9029 at the ICC, of whom 38.5% were female. Mean age was 37.4 ± 15.9 years. Incidences of previously undiagnosed TB (all forms) for SCC and ICC were 23.6 (95% CI 20.04-27.4) and 22.1 (95% CI 20.3-24.1) per 100,000 population ('P' = .42), SS+ TB 22.5 (95% CI 19.3-26.1) and 21.6 (95% CI 19.8-23.6) ('P' = .67), respectively. NNS to diagnose 1 TB case were 260 (95% CI 234.3-289.6) and 258 (95% CI 233.3-287.9) for SCC and ICC, respectively ('P' = .9). RRs for all forms of TB and SS+ TB in SCC compared to ICC were 0.94 (95% CI 0.73-1.19) and 0.95 (95% CI 0.74-1.22) and 'P' values were .58 and .71, respectively.ConclusionsBoth types of chest camps are equally effective in active case finding of previously undiagnosed TB cases in rural areas in 2 provinces in Pakistan.

A pituitary tumor is a common manifestation of the endocrinopathies criterion in McCune Albright syndrome. Herein, we report a 12-year-old male with McCune Albright syndrome, who presented with dysmorphic features and tall stature, then, underwent a trans-sphenoidal resection of suprasellar lesion that revealed a pituitary mammosomatotroph tumor with a unique finding of folliculo-stellate and mucin cells. Keywords: McCune Albright syndrome, Pituitary tumor, Mammosomatotroph, Folliculo-stellate cells

A critical challenge in scaffold design for tissue engineering is recapitulating the complex biochemical patterns that regulate cell behavior in vivo. In this work, we report the adaptation of a standard sterilization methodology—UV irradiation—for patterning the surfaces of two complementary polymeric electrospun scaffolds with oxygen cues able to efficiently immobilize biomolecules. Independently of the different polymer chain length of poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT) copolymers and PEOT/PBT ratio, it was possible to easily functionalize specific regions of the scaffolds by inducing an optimized and spatially controlled adsorption of proteins capable of boosting the adhesion and spreading of cells along the activated fibrous runways. By allowing an efficient design of cell attachment patterns without inducing any noticeable change on cell morphology nor on the integrity of the electrospun fibers, this procedure offers an affordable and resourceful approach to generate complex biochemical patterns that can decisively complement the functionality of the next generation of tissue engineering scaffolds.

Background: In Pakistan chronic obstructive pulmonary disease (COPD) prevalence is 2.1% in adults aged >40 years. Despite being a health policy focus, integrated COPD care has remained neglected, with wide variation in practice. Aim: To assess whether enhanced care at public health facilities resulted in better control of COPD, treatment adherence, and smoking cessation. Design & setting: A two-arm cluster randomised controlled trial was undertaken in 30 public health facilities (23 primary and 7 secondary), across three districts of Punjab, between October 2014–December 2016. Both arms had enhanced diagnosis and patient recording processes. Intervention facilities also had clinical care guides; drugs for COPD; patient education flipcharts; associated staff training; and mobile phone follow-up. Method: Facilities were randomised in a 1:1 ratio (sealed envelope independent lottery method), and 159 intervention and 154 control patients were recruited. The eligibility criteria were as follows: diagnosed with COPD, aged ≥18 years, and living in the catchment area. The primary outcome was change in BODE (Body mass index, airway Obstruction, Dyspnoea, Exercise capacity) index score from baseline to final follow-up visit. Staff and patients were not blinded. Results: Six-month primary outcomes were available for 147/159 (92.5%) intervention and 141/154 (91.6%) control participants (all clusters). The primary outcome results cluster-level analysis were as follows: mean intervention outcome = -1.67 (95% confidence intervals [CI] = -2.18 to -1.16); mean control outcome = -0.66 (95% CI = -1.09 to -0.22); and covariate-adjusted mean intervention–control difference = -0.96 (95% CI = -1.49 to -0.44; P = 0.001). Conclusion: The findings of this trial and a separate process evaluation study support the scaling of this integrated COPD care package at primary and secondary level public health facilities in Pakistan and similar settings.

Salt stress in soil is a critical constraint that affects the production of rice. Salt stress hinders plant growth through osmotic stress, ionic stress, and a hormonal imbalance (especially ethylene), therefore, thoughtful efforts are needed to devise salt tolerance management strategies. 1-Methylcyclopropene (1-MCP) is an ethylene action inhibitor, which could significantly reduce ethylene production in crops and fruits. However, 1-MCPs response to the physiological, biochemical and antioxidant features of rice under salt stress, are not clear. The present study analyzed whether 1-MCP could modulate salt tolerance for different rice cultivars. Pot culture experiments were conducted in a greenhouse in 2016–2017. Two rice cultivars, Nipponbare (NPBA) and Liangyoupeijiu (LYP9) were used in this trial. The salt stress included four salt levels, 0 g NaCl/kg dry soil (control, CK), 1.5 g NaCl/ kg dry soil (Low Salt stress, LS), 4.5 g NaCl/kg dry soil (Medium Salt stress, MS), and 7.5 g NaCl/kg dry soil (Heavy Salt stress, HS). Two 1-MCP levels, 0 g (CT) and 0.04 g/pot (1-MCP) were applied at the rice booting stage in 2016 and 2017. The results showed that applying 1-MCP significantly reduced ethylene production in rice spikelets from LYP9 and NPBA by 40.2 and 23.9% (CK), 44.3 and 28.6% (LS), 28 and 25.9% (MS), respectively. Rice seedlings for NPBA died under the HS level, while application of 1-MCP reduced the ethylene production in spikelets for LYP9 by 27.4% compared with those that received no 1-MCP treatment. Applying 1-MCP improved the photosynthesis rate and SPAD value in rice leaves for both cultivars. 1-MCP enhanced the superoxide dismutase production, protein synthesis, chlorophyll contents (chl a, b, carotenoids), and decreased malondialdehyde, H2O2, and proline accumulation in rice leaves. Application of 1-MCP also modulated the aboveground biomass, and grain yield for LYP9 and NPBA by 19.4 and 15.1% (CK), 30.3 and 24% (LS), 26.4 and 55.4% (MS), respectively, and 34.5% (HS) for LYP9 compared with those that received no 1-MCP treatment. However, LYP9 displayed a better tolerance than NPBA. The results revealed that 1-MCP could be employed to modulate physiology, biochemical, and antioxidant activities in rice plants, at different levels of salt stress, as a salt stress remedy.

Background: Hypertension in Pakistan affects 33% of people aged ≥45 years, and in urban areas around 70% of basic health care occurs in private facilities. Aim: To assess whether enhanced care at urban private clinics resulted in better control of hypertension, cardiovascular disease (CVD) risk factors, and treatment adherence. Design & setting: A two-arm cluster randomised controlled trial was conducted at 26 private clinics (in three districts of Punjab) between January 2015–September 2016. Both arms had enhanced screening and diagnosis of hypertension and related conditions, and patient recording processes. Intervention facilities also had a clinical care guide, additional drugs for hypertension, a patient lifestyle education flipchart, associated training, and mobile phone follow-up. Method: Clinics were randomised in a 1:1 ratio (sealed envelope lottery method). A total of 574 intervention and 564 control patients in 13 clusters in each arm were recruited (male and female, aged ≥25 years, systolic blood pressure [SBP] >140 mmHg, and/or diastolic blood pressure [DBP] >90 mmHg). The primary outcome was change in SBP from baseline to 9-month follow-up. Staff and patients were not blinded, but outcome assessors were blinded. Results: Nine-month primary outcomes were available for 522/574 (90.9%) intervention and 484/564 (85.8%) control participants (all clusters). The unadjusted cluster-level analysis results were as follows: mean intervention outcome was -25.2 mmHg (95% confidence intervals [CI] = -29.9 to -20.6); mean control outcome was -9.4 mmHg (95% CI = 21.2 to 2.2); and mean control–intervention difference was 15.8 (95% CI = 3.6 to 28.0; P = 0.01). Conclusion: The findings and separate process evaluation support the scaling of an integrated CVD–hypertension care intervention in urban private clinics in areas lacking public primary care in Pakistan.

Background: There were an estimated 7 million people living with diabetes in Pakistan in 2014, and this is predicted to reach 11.4 million by 2030. Aim: To assess if an integrated care package can achieve better control of diabetes. Design & setting: The pragmatic cluster randomised controlled trial (cRCT) was conducted from December 2014–June 2016 at 14 primary healthcare facilities in Sargodha district. Opportunistic screening, diagnostic testing, and patient recording processes were introduced in both the control 'testing, treating, and recording' (TTR) arm, and the intervention 'additional case management' (ACM) arm, which also included a clinical care guide and pictorial flipbook for lifestyle education, associated clinician training, and mobile phone follow-up. Method: Clinics were randomised on a 1:1 basis (sealed envelope lottery method) and 250 patients recruited in the ACM arm and 245 in the TTR-only arm (age ≥25 years and HbA1c >7%). The primary outcome was mean change in HbA1c (%) from baseline to 9-month follow-up. Patients and staff were not blinded. Results: The primary outcome was available for n = 238/250 (95.2%) participants in the ACM arm and n = 219/245 (89.4%) participants in the TTR-only arm (all clusters). Cluster level mean outcome was -2.26 pp (95% confidence intervals [CI] = -2.99 to -1.53) for the ACM arm, and -1.44 pp (95% CI = -2.34 to -0.54) for the TTR-only arm. Cluster level mean ACM–TTR difference (covariate-unadjusted) was -0.82 pp (95% CI = -1.86 to 0.21; P = 0.11). Conclusion: The ACM intervention in public healthcare facilities did not show a statistically significant effect on HbA1c reduction compared to the control (TTR-only) arm. Future evaluation should assess changes after a longer follow-up period, and minimal care enhancement in the comparator (control) arm.

The sudden breakout of coronavirus disease (COVID-19) rapidly spread across the globe, leaving no country behind in being affected by the global pandemic in the year 2019–20. As COVID-19 commenced, within months two major Asian giants initiated the norms of social distancing and lockdowns in their societies. The indiscriminate nature of the current pandemic has not only impacted the health and quality of life of people but has also disrupted the global economy, supply chains, and countries all over the world. In food and beverage manufacturing industries, the unanticipated disruption has encumbered its lock on the global food supply chain and service sector as major cities shut down for several months in China and India. Human existence is dependent upon food, which renders energy for activity, growth, and all functions of the body. Although both China and India have shown eminent response to tackle the ongoing pandemic, the food supply chain remains vastly exposed to significant COVID-19 risks. This research primarily investigates the ongoing COVID-19 scenario in two major economies (China and India), delivering insight into the pandemic’s impact within the food and beverage manufacturing sectors, and explores the policies adopted and strategies undertaken for sustainability in food supply chains.

Brevicoryne brassicae is a problematic pest in cabbage and other field crops. Synthetic pesticides are used to control this pest, but they are injurious for human health and the environment. The present study aimed to purify and identify the active compounds from Citrullus colocynthis leaves with an appraisal of their efficacy against B. brassicae. Separation and purification were performed via different chromatographic techniques. Molecular analysis and chemical structures were recognized by mass spectrum (MS) and nuclear magnetic resonance (NMR), respectively. Moreover, in vitro and in vivo aphicidal activity was assessed using various concentrations, i.e., 6.25, 12.5, 25 and 50 µg/mL at 12, 24, 48 and 72 h exposure. The outcome shows that mass spectrum analyses of the purified compounds suggested the molecular formulae are C30H50O and C29H50O, C29H48O. The compounds were characterized as fernenol and a mixture of spinasterol, 22,23-dihydrospinasterol by 1H-NMR and 13C-NMR spectrum analysis. The toxicity results showed that the mixture of spinasterol and 22,23-dihydrospinasterol showed LC50 values of 32.36, 44.49 and 37.50 µg/mL by contact, residual and greenhouse assay at 72 h exposure, respectively. In contrast, fernenol recorded LC50 values as 47.99, 57.46 and 58.67 µg/mL, respectively. On the other hand, spinasterol, 22,23-dihydrospinasterol showed the highest mortality, i.e., 66.67%, 53.33% and 60% while, 30%, 23.33% and 25% mortality was recorded by fernenol after 72 h at 50 µg/mL by contact, residual and greenhouse assay, respectively. This study suggests that spinasterol, 22,23-dihydrospinasterol are more effective against B. brassicae which may be introduced as an effective and suitable substitute of synthetic chemical pesticides.

Rice (Oryza sativa L.) is a major cereal food crop worldwide, and its growth and yield are affected by several fungal phytopathogens, including Magnaporthe oryzae, Fusarium graminearum, F. moniliforme, and Rhizoctonia solani. In the present study, we have isolated and characterized root-associated bacterial endophytes that have antifungal activities against rice fungal phytopathogens. A total of 122 root-associated bacterial endophytes, belonging to six genera (Bacillus, Fictibacillus, Lysinibacillus, Paenibacillus, Cupriavidus, and Microbacterium) and 22 species were isolated from three rice cultivars. Furthermore, the 16S rRNA sequence-based phylogeny results revealed that Bacillus was the most dominant bacterial genera, and that there were 15 different species among the isolates. Moreover, 71 root-associated endophytes showed antagonistic effects against four major fungal phytopathogens, including M. oryzae, F. graminearum, F. moniliforme, and R. solani. Additionally, the biochemical, physiological, and PCR amplification results of the antibiotic-related genes further supported the endophytes as potential biocontrolling agents against the rice fungal pathogens. Consequently, the findings in this study suggested that the isolated bacterial endophytes might have beneficial roles in rice defense responses, including several bioactive compound syntheses. The outcomes of this study advocate the use of natural endophytes as an alternative strategy towards the rice resistance response.

Background: Sophisticated health education is now become a prime requirement to bring improvements in health profiles of patients and their families. Health care activities play a significant role to keep health education at forefront. The aim of the present study isto determine the health care workers’ perception for the need of instituting health promotion and also identify their beliefs about the improvement of community health by conducting health literacy sessions. Methods: A cross sectional study was conducted from April to October 2016 in various districts of Karachi. A total 286 medical care providers including physicians, pharmacists, nurses, dispensers/compounders, academicians and medical officers/quacks belonging to different organizations/institutions were approached.A structured questionnaire was developed containing different close ended items, seems to be necessary to meet objective of the study. Results: About 82.42% of them felt the need of health promotion services owing to their belief that such counseling would be beneficial in maintaining better health status of population. The lack of information was seen regarding drug-drug (24.12%) and food-drug interactions (25.87%). The most common issue, addressed by health care providers (68.53%) was the improvement in physical fitness via life style modification. The healthcare providers utilized various ways for spreading of information; however the role of media was highly encouraged by them. Conclusion: Institution of health education programs for residents is indeed necessary to reduce the risks of many communicable and non-communicable diseases.Therefore, such activities should be planned on regular basisin various areas of the city to provide maximum opportunitiesto individuals for participation. Health counseling also played a vital role and considered to be beneficial in the development of healthy community/society.

Background: Respiratory diseases, namely asthma and chronic obstructive pulmonary disease (COPD), account for one-fourth of the patients at the primary health-care (PHC) facilities in Pakistan. Standard care practices to manage these diseases are necessary to reduce the morbidity and mortality rate associated with non-communicable diseases in developing countries. Objective: To develop and measure the effectiveness of operational guidelines and implementation materials, with sound scientific evidence, for expanding lung health care, especially asthma and COPD through PHC facilities already strengthened for tuberculosis (TB) care in Pakistan. Design: A cluster randomized controlled trial with two arms (intervention and control), with qualitative and costing study components, is being conducted in 34 clusters; 17 clusters per arm (428 asthma and 306 COPD patients), in three districts in Pakistan from October 2014 to December 2016. The intervention consists of enhanced case management of asthma and COPD patients through strengthening of PHC facilities. The main outcomes to be measured are asthma and COPD control among the registered cases at 6 months. Cluster- and individual-level analyses will be done according to intention to treat. Residual confounding will be addressed by multivariable logistic and linear regression models for asthma and COPD control, respectively. The trial is registered with ISRCTN registry (ISRCTN 17409338). Conclusions: Currently, only about 20% of the estimated prevalent asthma and COPD cases are being identified and reported through the respective PHC network. Lung health care and prevention has not been effectively integrated into the core PHC package, although a very well-functioning TB program exists at the PHC level. Inclusion of these diseases in the already existent TB program is expected to increase detection rates and care for asthma and COPD.

Objective: Sexually transmitted infections (STIs) are a priority health problem. We proposed a prospective study in two districts of Punjab, using an intervention package, which included guidelines and protocols on syndrome-based management of STIs, adapted in light of technical guidelines from the National AIDS Control Program and the World Health Organization. The aim of this study was to assess the operational effectiveness of STI case management guidelines and to assess factors that determine the adherence to guidelines for management of STIs at public health facilities in Pakistan. Methods: A prospective study lasting 18 months (January 2008 to June 2009), which reviewed early implementation experiences of updated case management guidelines for delivery of syndrome-based STI/reproductive tract infection care, through public-sector health care facilities. The project was implemented in two districts of Punjab, Sargodha and Jhang. A Cox regression model with stratification was done. Findings: The prevalence of STI was 26 per 100,000 patients. In women, the reported symptoms were 80% vaginal discharge and 12% abdominal pain. Forty-four percent of men had a genital ulcer and 29% of men had genital discharge. Age of participants ranged from 13 to 60 years. The study comprised 28.6% men and 71.4% women. The majority of the population attending these clinics was from rural areas (70%). The variables independently associated with adherence to guidelines were availability of male paramedic, age of patient, and type of diagnosis made. There was an important interaction (effect modification) present between the area of health facility and patient sex. Conclusion: Screening, diagnosis, and treatment costs for many STIs are expensive and thus an easier, low-cost, syndrome-based public health strategy is the adoption of the proposed STI syndrome case management guidelines.

Owing to the pharmacological potential of ATRA (all trans-retinoic acid), a series of retinamides and a 1-(retinoyl)-1,3-dicyclohexylurea compound were prepared by reacting ATRA with long chain alkyl or alkenyl fatty amines by using a 4-demethylaminopyridine (DMAP)-catalyzed N,N¢-dicyclohexylcarbodiimide (DCC) coupling. The successful synthesis of the target compounds was demonstrated using a range of spectroscopic techniques. The cytotoxicity of the compounds was measured along with their ability to induce cell cycle arrest and apoptosis in human cancer cell lines MCF-7 (breast cancer) and HepG2 (liver cancer) and normal human cell line HEK293 (embryonic kidney). The results of cytotoxicity and flow cytometry data showed that the compounds had a moderate to strong effect against MCF-7 and HepG2 cells and were less toxic to HEK293 cells. N-oleyl-retinamide was found to be the most potent anticancer agent and was more effective against MCF-7 cells than HepG2 cells.

Aim: To optimize the concentrations of growth regulators in the media for the proficient micropropagation of grapevine (Vitis vinifera L.) cv. King’s Ruby. Methods and results: Apical meristems of the grape cultivar were used to establish in vitro shoot cultures. Nodal explants, each containing an axillary bud, taken from in vitro grown shoots were inoculated in shoot proliferation medium, i.e., half strength Murashige and Skoog (MS) medium supplemented with benzyl aminopurine (BAP), kinetin, glycine and gibberellic acid (GA3). A higher number of shoots (5.33) with greater shoot length (2.75 cm) was produced in the medium supplemented with 1.0 mg L-1 BAP and 0.1 mg L-1 GA3. Calluses were induced from leaf explants taken from in vitro grown shoots. Callus induction was greater (73.00%) on the medium containing 2.0 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.3 mg L-1 BAP and 0.2 mg L-1 α-naphthaleneacetic acid (NAA). The maximum frequency of shoot regeneration (53.33%) was achieved on the medium supplemented with 1.5 mg L-1 BAP and 0.5 mg L-1 NAA, and the regenerated shoots successfully formed roots on growth regulator-free half strength MS medium. Conclusion: Optimizing the concentration of BAP and GA3 and omitting the glycine and kinetin in the culture medium increased the number and length of shoots. Similarly, for inducing the callus of the leaf explants, taken from in vitro grown shoots, it is recommended to adjust the medium with the higher concentration of 2,4-D and lower concentrations of BAP. Moreover, the maximum number of shoots was regenerated on a medium supplemented with relatively high levels of both BAP and NAA (1.5 and 0.5 mg L-1, respectively). Finally, we suggest the half strength MS medium that is free from growth regulators for the root formation of the regenerated shoots. Significance and impact of the study: Optimizing the concentration of growth regulators is crucial for the efficient micropropagation of a grape cultivar. Knowing the specific balance between the growth regulators is necessary to establish in vitro shoot cultures, callus induction and shoot regeneration and, hence, to propagate disease-free true to type grape cultivars in a short time.