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Background The preparation of an effective drug delivery formulation is an urgent need to treat cutaneous leishmaniasis (CL). Pentavalent antimonials and Amphotericin B (AmB) are considered to treat leishmaniasis; however, their clinical usage is hampered by poor solubility, high cost, toxicity, and the emergence of drug-resistant Leishmania spp. The drug delivery systems (DDS) could be used as an alternative treatment option for the treatment of CL to circumvent these problems. We tested the antileishmanial efficacies of free AmB and amphotericin B-loaded chitosan nanoparticles (AmB-CNPs) under in vitro conditions. Results Chitosan nanoparticles (CNPs) were synthesized using the ionic gelation method with negatively charged tripolyphosphate (TPP). During the synthesis of CNPs, AmB was incorporated into the nanoparticles (NPs). The NPs were characterized for their size, surface morphology, encapsulation efficacy (EE), drug loading content (DLC), and surface charge using different techniques. Their efficacy was evaluated against promastigotes and axenic amastigotes forms of Leishmania tropica using MTT assay. The synthesized AmB-CNPs displayed a spherical shape with a mean particle size of 118 nm, a positive zeta potential of (+ 6.21 ± 2.02 mV), and an encapsulation efficacy of 88%. Dynamic light scattering technique (DLS) shows that the average size of prepared AmB-CNPs was 95.5 nm. Free AmB presented very low efficacy (only 65% and 67% inhibition of the promastigotes and axenic amastigotes parasite load), whereas AmB-CNPs exhibited 90% and 84% parasite inhibition after 72 h incubation. The AmB-CNPs exhibited significantly higher efficacy than free AmB in terms of reduction in parasite viability. Half-maximal inhibitory concentration (IC50) measured values of the AmB-CNPs were significant lowers than free AmB. Conclusions The present data indicated that AmB-CNPs exhibited vigorous anti-leishmanial activity than free AmB by dose and time-dependent manner. This formulation can be used for local therapy of CL after in vivo efficacy conformational studies.

Several archaea harbour genes that code for fructosyltransferase (FTF) enzymes. These enzymes have not been characterized yet at structure-function level, but are of extreme interest in view of their potential role in the synthesis of novel compounds for food, nutrition and pharmaceutical applications. In this study, 3D structure of an inulin-type fructan producing enzyme, inulosucrase (InuHj), from the archaeon Halalkalicoccus jeotgali was resolved in its apo form as well as with bound substrate (sucrose) molecule and first transglycosylation product (1-kestose). This is the first crystal structure of an FTF from halophilic archaea. Its overall five-bladed β-propeller fold is conserved with previously reported FTFs, but also shows some unique features. The InuHj structure is closer to those of Gram-negative bacteria, with exceptions such as residue E266, which is conserved in FTFs of Gram-positive bacteria and has possible role in fructan polymer synthesis in these bacteria as compared to fructooligosaccharide (FOS) production by FTFs of Gram-negative bacteria. Highly negative electrostatic surface potential of InuHj, due to a large amount of acidic residues, likely contributes to its halophilicity. The complex of InuHj with 1-kestose indicates that the residues D287 in the 4B-4C loop, Y330 in 4D-5A and D361 in the unique α2 helix may interact with longer FOSs and facilitate the binding of longer FOS chains during synthesis. The outcome of this work will provide targets for future structure-function studies of FTF enzymes, particularly those from archaea.

Tomato plant (Lycopersicon esculentum) is a widely cultivated vegetable crop. The plant is economically very important. To ensure a successful and economically viable harvest, it is essential that all aspects of its production to be monitored. At present, there is a limited scientific data regarding the use of pesticides in relation to the seasonal incidence of insect pests in the Bajaur district of Pakistan. This study was carried out at a local farm, where the efficacies of Flurofenafire pesticide were examined. The pesticide Flurofenafire was applied once a week to examine its efficacy. The stability of the pesticide was highly dependent on the weather. The occurrence of rain within 24 hours of spraying resulted in Aphid, Jassid, White fly, Thrips and Fruit borer infestations. Aphid infestations were observed on the tomato crop in May 2019, which peaked in July. Bemisia Tabaci infestations peaked in August. Amrasca Biguttula populations peaked in September 2019. Scirtothrips dorsalis and Helicoverpaarmigera infestations were also observed on the tomato crops and caused significant damage. The application of Flurofenafire was able to control these infestations. In conclusion the pesticide Flurofenafire was found effective against a wide range of insect pest of tomato plant. Key Words: Flurofenafire, Pest, Pesticide, Seasonal incidence, Lycopersicon esculentum

Particle-induced X-ray emission (PIXE) method was used in this present research to identify the elements present in selected vegetable samples to show the possible influence in the metal absorption by the vegetables grown in a saline region of Rampal area of Munshiganj District, Bangladesh. The data acquisition setup is calibrated using a 2.5-MeV proton beam in the current ranges of 5nA to 15nA. The detector was used to measure the X-rays emitted during the irradiation. Data acquisition system MAESTRO-32 was used to measure the spectrum picks, and concentration calculation has been done by GUPIX/DAN-32 software. The purpose of this study is to determine the concentration of heavy and trace elements in these samples and to give current information on their safety for consumption. The result shows that K, Ca, and Fe have the highest concentrations, while lead exhibits the lowest but alarming rates compared to reference materials. The findings were likened to IAEA-V-10, IAEA-359, SRM 1515 (apple leaf), and SRM 1573a (tomato leaf). The outcomes of the present investigation demonstrate that these samples are not devoid of health risks in intake.

This study is based on the analysis of language practices and features that different cricket commentators use in international cricket matches. This study explores that cricket commentary forms a complete register on its own with its field, tenor, and mode components. Data for this research have been taken from the 1992 World Cup, the 2015 World Cup, and the Ashes Series 2005 matches available on YouTube. Textual analysis technique has been employed for data analysis. After analyzing the data in the light of Halliday’s notion of register comprising field, tenor, and mode, it is found that cricket commentary is characterized by special syntactic features such as ellipsis, extensive use of exclamatory sentences, the abundant use of the simple present tense and simple sentences, passive construction notably in elliptical form, first and third conditionals, and inversion. Use of idioms, certain action verbs, certain positive and negative adjectives, and certain adverbs are some of the salient features of cricket commentary. Cricket commentary also has a special jargon with words like ‘knock’, ‘mid-on’, ‘mid-off’, ‘innings’ and ‘bouncer’ etc. Cricket commentators also use certain aspects of connected speech such as weak forms of words, elision, and assimilation.

Aim: To evaluate antimicrobial activity of extracellular metabolites (EMs) of endophytic fungal isolates (EFIs) from Azadirachta indica. Materials & methods: EFIs were identified by internal transcribed spacer (ITS) sequencing. Antimicrobial activity, and minimum inhibitor concentration (MIC) and minimum bactericidal concentration (MBC) were determined using agar diffusion and microdilution method, respectively. Results: Seventeen EFIs were isolated from different organs of A. indica. Eight of them were identified based on ITS sequencing. The EMs of EFIs inhibited the growth of six multidrug-resistant (MDR) bacterial superbugs and three phytopathogenic fungi. The MDR bacterial superbugs are resistant to six commercial antibiotics of different generations but susceptible to EMs of EFIs. The MIC (0.125–1.0 μg/μl), MBC (0.5–4.0 μg/μl) and minimum fungicidal concentration (1.0–4.0 μg/μl) of the EMs from EFIs are lower enough. Conclusion: The EMs of the EFIs have promising antimicrobial activity against MDR bacteria and phytopathogenic fungi.

Introduction: Enterobacteriaceae such as Escherichia coli and Klebsiella pneumoniae are the most prominent bacterial species resistant to almost all commonly used antibiotics. Carbapenem is one of the last resort drugs for treating such emerging multidrug-resistant bacteria. This study aimed to detect carbapenem-resistant blaNDM-1 gene in ESBL producing E. coli and K. pneumoniae isolates. Methodology: A total of 190 E. coli and 350 K. pneumoniae isolates were screened for extended spectrumβ-lactamase (ESBL), carbapenemase and metallo β-lactamase (MBL) production via double-disk synergy test (DDST), modified Hodge test and combined-disk diffusion method. The blaNDM-1 gene was detected by PCR and confirmed via Sanger sequencing method. Results: Of the 540 isolates tested, 71.8% were found to be multidrug-resistant. Overall rate of ESBL-positive isolates were 57.89% E. coli and 31.42% K. pneumoniae. Among ESBL positive isolates, 49.09% E. coli and 40% K. pneumoniae were positive for carbapenemase production whereas MBL production was detected in 29% E. coli and 22% K. pneumoniae isolates. In MBL positive isolates, (37%) E. coli and (40%) K. pneumoniae isolates harboured blaNDM-1 gene. The pair-wise DNA was aligned with the NDM-1 sequence from GenBank. The alignment score was 243 and the blast nucleotide sequencing results showed 97% sequence similarity with the sequences in GenBank for the blaNDM-1 gene. Conclusions: The blaNDM-1 gene was found to be the most prevalent in urine samples. There is a dire need to conduct screening tests in hospitals and communities to find out the exact prevalence of the blaNDM-1 spread in our population.

Vacuolar H+-ATPases (V-ATPases) transport protons across cellular membranes to acidify various organelles. ATP6V0A1 encodes the a1-subunit of the V0 domain of V-ATPases, which is strongly expressed in neurons. However, its role in brain development is unknown. Here we report four individuals with developmental and epileptic encephalopathy with ATP6V0A1 variants: two individuals with a de novo missense variant (R741Q) and the other two individuals with biallelic variants comprising one almost complete loss-of-function variant and one missense variant (A512P and N534D). Lysosomal acidification is significantly impaired in cell lines expressing three missense ATP6V0A1 mutants. Homozygous mutant mice harboring human R741Q (Atp6v0a1R741Q) and A512P (Atp6v0a1A512P) variants show embryonic lethality and early postnatal mortality, respectively, suggesting that R741Q affects V-ATPase function more severely. Lysosomal dysfunction resulting in cell death, accumulated autophagosomes and lysosomes, reduced mTORC1 signaling and synaptic connectivity, and lowered neurotransmitter contents of synaptic vesicles are observed in the brains of Atp6v0a1A512P/A512P mice. These findings demonstrate the essential roles of ATP6V0A1/Atp6v0a1 in neuronal development in terms of integrity and connectivity of neurons in both humans and mice.

Hazrat Bilal,1 Tayyab Ur Rehman,2 Muhammad Asif Khan,2 Fareeha Hameed,2 Zhang Gao Jian,1 Jianxiong Han,1 Xingyuan Yang1 1Faculty of Health Sciences, Institute of Physical Sciences and Information Technology, Anhui University, Hefei, People’s Republic of China; 2Institute of Basic Medical Sciences, Khyber Medical University, Peshawar, PakistanCorrespondence: Xingyuan YangFaculty of Health Sciences, Institute of Physical Science and Information Technology, Anhui University, No, 111 Jiulong Road, Hefei, Anhui, 230601, People’s Republic of ChinaEmail xingyuan@ahu.edu.cnPurpose: The multiple-drug resistant Escherichia coli are among the deadliest pathogens causing life-threatening infections. This study was planned to determine the molecular epidemiology of mcr-1, blaKPC-2, and blaNDM-1 harboring clinically isolated E. coli from Pakistan.Methods: In total, 545 strains of E. coli from clinical samples were collected from June 2018 to September 2019. All the isolates were screened for colistin-resistance, extended-spectrum-β-lactamases (ESBL), and carbapenemases through the micro-dilution method, Double-Disk-Synergy-Test (DDST), and Modified-Hodge-Test (MHT). The detection, sequence-typing, conjugal transfer, S1-PFGE, plasmid-replicon-typing, and southern-blotting for mcr, ESBL, and carbapenemase-encoding genes were performed.Findings: A total of four (0.73%) colistin-resistant strains carrying alongside mcr-1 and blaCTX-M-15 genes, three of these strains also had the blaTEM-1 gene. The presence of ESBL genes was detected in 139 (25.5%) isolates harboring blaCTXM-15 (74.82%), blaTEM (34.53%), blaSHV (28.06%) and blaOXA-1 (28.78%). In 129 carbapenemase-producers, 35.83% possessed blaNDM-1, 26.67% blaKPC-2, 8.3% blaOXA-48, 25% blaVIM-1, and 20.83% blaIMP-1 genes. The sequence typing revealed that mcr-1 harboring isolates belonged to ST405, ST117, and ST156. Fifty percent of blaKPC-2 and 48.83% of blaNDM-1 were found on ST131 and ST1196, respectively. Two rare types of STs, ST7584, and ST8671 were also identified in this study. The mcr-1 gene was located on Incl2 (60-kb) plasmid. The blaKPC-2 was present on (140-kb) IncH12, (100-kb) IncN, (90-kb) Incl1, while blaNDM-1 was located on (70-kb) IncFIIK, (140-kb) IncH12, (100-kb) IncN, (60-kb) IncA/C, and (45-kb) IncFII plasmids, which were successfully trans-conjugated. Among the plasmid types, the Incl1 carrying blaKPC-2, IncH12 harboring blaKPC-2 and blaNDM-1, and IncFIIK carrying blaNDM-1 were for the first time detected in Pakistan.Conclusion: The mcr-1, blaKPC-2, and blaNDM-1 genes finding in various clonal and plasmids types indicate that a substantial selection of the resistance genes had occurred in our clinical strains.Keywords: colistin-resistance, extended-spectrum-β-lactamases, carbapenemases, mcr1, blaKPC-2, blaNDM-1, plasmids, E. coli

Muhammad Fazal Hameed,1 Yanan Chen,1 Ying Wang,2 Muhammad Shafiq,3 Hazrat Bilal,1 Linqing Liu,4 Jinming Ma,1 Pengying Gu,4 Honghua Ge1 1Institutes of Physical Science and Information Technology, Anhui University, Hefei, Anhui, 230601, People’s Republic of China; 2Department of Laboratory Medicine, The First Affiliated Hospital of USTC, Division of Life Science and Medicine, University of Science and Technology of China, Hefei, Anhui, 230036, People’s Republic of China; 3College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China; 4The Department of Geriatrics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230036, People’s Republic of ChinaCorrespondence: Honghua GeInstitutes of Physical Science and Information Technology, Anhui University, Hefei, Anhui, 230601, People’s Republic of ChinaTel +86-551-63861773Email hhge@ahu.edu.cnPengying GuThe Department of Geriatrics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230036, People’s Republic of ChinaEmail hellengpy@163.comPurpose: Antimicrobial resistance, especially carbapenem resistance Enterobacteriaceae and plasmid mediated mobile colistin resistance, is a serious issue worldwide. This study was designed to determine the epidemiological characteristics of plasmid mediated colistin resistance and carbapenem resistant Enterobacteriaceae from tertiary A hospital located in Hefei, China.Methods: Totally, 158 carbapenems resistant Enterobacteriaceae (CRE) were screened for antibiotic susceptibility, mcr-1, extended spectrum β-lactamases (ESBLs), metallo-β-lactamases (MBLs), and fosfomycin resistance genes using PCR and sequencing. The sequence types were identified by multilocus sequence typing (MLST). Plasmid profiles were determined by PCR based replicon typing (PBRT), and the plasmid sizes were confirmed by southern blotting.Results: The isolates showed high MIC50 and MIC90 for all antimicrobials, except tigecycline, meropenem, and colistin. The main Carbapenemase genes were blaKPC-2 (90.5%), blaNDM-1(3.7%), blaOXA-48(5.6%) and fosA3 (14.5%). The blaCTXM-15 found 36.7%, mcr-1 (3.7%) recorded in six isolates. PBRT revealed blaKPC-2 in K. pneumoniae on IncR, IncFII, and IncA/C. blaNDM-1 in E. coli on IncFII, whereas in E. cloacae noticed on IncHI2 plasmid. mcr-1 was recorded among IncFIIK, IncFII, and IncF in E. coli, K. pneumoniae, and E. cloacae. Resistance genes (mcr-1, blaNDM-1, blaKPC-2) harboring plasmids are successfully trans-conjugant to EC-600. A high incidence of ST11 was observed in K. pneumoniae carbapenem resistant isolates. While in E. coli, multiple STs were identified. However, mcr-1 in ST23 was identified for the first time in Anhui Province. Among Enterobacter cloacae, ST270 detected carrying blaNDM-1. Southern-hybridization confirmed the plasmid sizes 35– 150kb.Conclusion: This study indicates the co-carrying of mcr-1, blaKPC-2, and blaNDM-1 among clinical isolates, the prevalence of different Enterobacteriaceae STs is alarming, especially in E. coli. Holding such a resistance profile is a threat for humans and animals, which may be transferred between the strains through plasmid transfusion. Persistent control actions are immediately necessary to combat this hazard.Keywords: ESBL, MBL, KPC2, mcr-1, NDM-1, CTXM-15

Background During the last six decades, extensive use of antibiotics has selected resistant strains, increasing the rate of fatal infectious diseases, and exerting an economic burden on society. This situation is widely accepted as a global problem, yet its degree is not well elucidated in many regions of the world. Up till now, no systemic analysis of Antimicrobial resistance (AMR) in Pakistan has been published. The current study aims to describe the antibiotic-resistance scenario of Pakistan from human samples of the last 10 y, to find the gaps in surveillances and methodology and recommendations for researchers and prescribers founded on these outcomes. Methods Original research articles analyzed the pattern of Antibiotic resistance of any World Health Organization (WHO) enlisted priority pathogens in Pakistan (published onward 2009 till March 2020), were collected from PubMed, Google scholar, and PakMedi Net search engines. These articles were selected based on predefined inclusion and exclusion criteria. Data about the study characteristics and antibiotic-resistance for a given bacterium were excluded from literature. Antibiotic resistance to a particular bacterium was calculated as a median resistance with 95% Confidence Interval (CI). Results Studies published in the last 10 y showed that Urinary Tract Infection (UTI) is the most reported clinical diagnosis (16.1%) in Pakistan. E. coli were reported in 28 (30.11%) studies showing high resistance to antibiotics’ first line. Methicillin-resistant Staphylococcus aureus (MRSA) was found in 49% of S. aureus’ total reported cases. Phenotypic resistance pattern has mostly been evaluated by Disk Diffusion Method (DDM) (82.8%), taken Clinical Laboratory Standards Institute (CLSI) as a breakpoint reference guideline (in 79.6% studies). Only 28 (30.11%) studies have made molecular identification of the resistance gene. blaTEM (78.94% in Shigella spp) and blaNDM-1 (32.75% in Klebsiella spp) are the prominent reported resistant genes followed by VanA (45.53% in Enterococcus spp), mcr-1 (1.61% in Acinetobacter spp), and blaKPC-2 (31.67% in E. coli). Most of the studies were from Sindh (40.86%), followed by Punjab (35.48%), while Baluchistan’s AMR data was not available. Conclusion Outcomes of our study emphasize that most of the pathogens show high resistance to commonly used antibiotics; also, we find gaps in surveillances and breaches in methodological data. Based on these findings, we recommend the regularization of surveillance practice and precise actions to combat the region’s AMR.

The current study investigates the toxic effects of acute dose of an endocrine disruptor atrazine on Grass carp (Ctenepharyngodon idella) using histopathological changes as biomarkers. Histopathology is promising field for research in aquatic toxicology, in this manner vital organs; brain, gills, and liver tissues were inspected histological after exposing to sublethal groupings of atrazine 0.025 and 0.03 μl/L for 3, 6, 9, 12 and 15 days individually with equal untreated control group. Against various doses, rapid movements, gulping of air and jumping of fish to scat from toxic medium were noticed. Various severe (+++) morphological modifications in tissue were documented in comparison with control group comprised of degenerated neurons, vacuolization, inflammatory cells infiltration and neural necrosis in brain tissue. The most well-known gills tissue alteration at all concentrations of atrazine was epithelial hyperplasia, desquamation, epithelial lifting and smaller aneurism while hepatic injuries were described by overcast expanding of hepatocytes esteemed as cloudy swelling of hepatocytes followed by karyolysis, karyohexis and dilation of sinusoids which shows that atrazine introduction upgraded the toxicosis impacts with the increase concentration, influenced the strength of the fish, inferable from histological inconsistencies.

Repeatedly deposition of anthropogenic chemicals such as chlorpyrifos led to the risk of its associated unwanted toxicity to the non-targeted organisms. The present project was designed to assess the toxicity of chlorpyrifos, an organophosphorus insecticide, at different dose level and duration of exposure to elucidate its cellular and nuclear anomalies in erythrocytes and histopathological effect on brain and liver tissues of the fresh water fish (Ctenopharyngodon idella). The chlorpyrifos was used at 0.01 and 0.015 μL/L for a duration of 3, 6, 9, 12, and 15 days respectively. Alteration in functional markers such as cellular anomalies associated with exposed dosage was reported as swollen cells, swollen or enlarged cells, and hemolyzed cells followed by nuclear abnormalities stated as micronucleus, deformed nucleus and nuclear shift. The result indicated alterations observed in brain and liver tissues were categorized as necrosis, vacuolization, degeneration of neurons and hepatic necrosis, hepatolysis and hepatic vacuolization that were further scored on the basis of severity from 0 (normal) to 3 (severe). These effects significantly increased with the increase in dose level of chlorpyrifos and duration of exposure (p < 0.05). The present findings contributed that extended exposure of such chemicals to the aquatic ecosystems led to behavioral and histopathological alterations that may result in loss of normal functioning of the organs.

Honey is a suitable matrix for the evaluation of environmental contaminants including organochlorine insecticides. The present study was conducted to evaluate residues of fifteen organochlorine insecticides in honey samples of unifloral and multifloral origins from Dir, Pakistan. Honey samples (5 g each) were extracted with GC grade organic solvents and then subjected to Rotary Evaporator till dryness. The extracts were then mixed with n-Hexane (5 ml) and purified through Column Chromatography. Purified extracts (1μl each) were processed through Gas Chromatograph coupled with Electron Capture Detector (GC-ECD) for identification and quantification of the insecticides. Of the 15 insecticides tested, 46.7% were detected while 53.3% were not detected in the honey samples. Heptachlor was the most prevalent insecticide with a mean level of 0.0018 mg/kg detected in 80% of the samples followed by β-HCH with a mean level of 0.0016 mg/kg detected in 71.4% of the honey samples. Honey samples from Acacia modesta Wall. were 100% positive for Heptachlor with a mean level of 0.0048 mg/kg followed by β-HCH with a mean level of 0.003 mg/kg and frequency of 83.3%. Minimum levels of the tested insecticides were detected in the unifloral honey from Ziziphus jujuba Mill. Methoxychlor, Endosulfan, Endrin and metabolites of DDT were not detected in the studied honey samples. Some of the tested insecticides are banned in Pakistan but are still detected in honey samples indicating their use in the study area. The detected levels of all insecticides were below the Maximum Residue Levels (MRLs) and safe for consumers. However, the levels detected can cause mortality in insect fauna. The use of banned insecticides is one of the main factors responsible for the declining populations of important insect pollinators including honeybees. Resumo O mel é uma matriz adequada para a avaliação de contaminantes ambientais, incluindo inseticidas organoclorados. O presente estudo foi conduzido para avaliar resíduos de 15 inseticidas organoclorados em amostras de mel de origem unifloral e multifloral de Dir, Paquistão. Amostras de mel (5 g cada) foram extraídas com solventes orgânicos de grau GC e, em seguida, submetidas ao evaporador rotativo até a secura. Os extratos foram então misturados com n-hexano (5 ml) e purificados por cromatografia em coluna. Os extratos purificados (1μl cada) foram processados através de cromatógrafo gasoso acoplado a detector de captura de elétrons (GC-ECD) para identificação e quantificação dos inseticidas. Dos 15 inseticidas testados, 46,7% foram detectados enquanto 53,3% não foram detectados nas amostras de mel. O heptacloro foi o inseticida mais prevalente com um nível médio de 0,0018 mg / kg detectado em 80% das amostras, seguido por β-HCH com um nível médio de 0,0016 mg / kg detectado em 71,4% das amostras de mel. Amostras de mel da parede de Acacia modesta foram 100% positivos para heptacloro com um nível médio de 0,0048 mg / kg seguido por β-HCH com um nível médio de 0,003 mg / kg e frequência de 83,3%. Níveis mínimos dos inseticidas testados foram detectados no mel unifloral de Ziziphus jujuba da usina. Metoxicloro, Endosulfan, Endrin e metabólitos do DDT não foram detectados nas amostras de mel estudadas. Alguns dos inseticidas testados são proibidos no Paquistão, mas ainda são detectados em amostras de mel, indicando seu uso na área de estudo. Os níveis detectados de todos os inseticidas estavam abaixo dos Níveis Máximos de Resíduos (MRLs) e seguros para os consumidores. No entanto, os níveis detectados podem causar mortalidade na fauna de insetos. O uso de inseticidas proibidos é um dos principais fatores responsáveis pelo declínio das populações de importantes insetos polinizadores, incluindo as abelhas.

The present study aims to explore the larvicidal activity of n-hexane extract isolated fractions from Artemisia scoparia against third instar larvae of Culex quinquefasciatus mosquito. Twelve fractions, F1 to F12 were separated through column chromatography by using different eluent solvents such as hexane, hexane-chloroform, chloroform and chloroform-methanol, in the order of increasing polarity. Larval bioassay was done by using WHO standard guideline. Larvicidal potential of the separated fractions was evaluated at tested concentrations of 30, 60, 90 and 120 ppm. Mortality rate was scored 24 h post exposure. The fractions F3 and F4 were found more active, exhibiting LC50 values of 28.66 and 22.37 ppm and LC90 values of 53.59 and 53.16 ppm respectively. The larvicidal activity of different fractions was compared by using ANOVA analysis.

Background and Objective: The mucoviscosity associated gene A (magA) in the hypermucoviscous variants of K. pneumoniae is reported to be associated with invasive infections and considered a virulence factor. We sought to analyze the magA genes in K. pneumoniae isolates in the clinical specimen collected from Bangladesh. Methods: We established a multicenter cohort of patients with Klebsiella infection hospitalized at 05 different hospitals between September 2016 and April 2017. We collected 313 K. pneumoniae isolates from patients who consented to participate in the study. The isolates were evaluated for harboring the magA genes using a single-tube multiplexed polymerase chain reaction. The magA genes were analyzed by PCR-RFLP technique using two enzymes, namely PciI and SmaI. Antibiogram assay using 12 commercially available antibiotic discs was performed on all the isolates. Results: The presence of K. pneumoniae specific gene (ureD) was confirmed in all the isolates. The percentage of isolates harboring the magA gene was 7.34%(23 isolates), the majority of which was collected from the patients admitted in intensive care units (16 isolates, 69.6%), and infectious diseases wards (5 isolates, 21.7%). PCR-RFLP analysis revealed that for 7 out of 23 isolates, where Sma1 could not cleave the magA gene. All the isolates showed resistance to ampicillin, carbenicillin cefradine, chloramphenicol, erythromycin, kanamycin, and sulphamethoxazole, though the extent was varying. However, imipenem showed 100% sensitivity to all the tested isolates. Conclusion: This study demonstrates the presence of the magA gene in multidrug-resistant clinical isolates of K. pneumoniae collected from Bangladesh.

Fructose polysaccharides and oligosaccharides are gaining strong importance due to their beneficial effects on human health and their applications in the food, feed and pharmaceutical industries. F...

A total of 30 plant species belonging to 14 genera of Rosaceae were collected from different localities of Dir upper. In which seven plant species were belonging to genus Prunus, five Rosa, three of Potentilla, two each of Cotoneaster, Rubus and Pyrus, one each of Crataegus, Eriobotrya, Fragaria, Malus, Sibbaldia, Sorbus, Sorbaria, Spiraea. It was concluded that members of the Rosaceae need attention not only for its cultivation but emphasis must be made on conservation for sustainable utilization of plant resources.

In developing countries, Cd contamination is ubiquitous which limits agriculture productivity. The current study was designed to investigate the efficacy of plant—Bacillus pumilus—ethylene diamine ...

Anthropogenic activities in the past and present eras have created global warming and consequently a storm of drought stress, affecting both plants and animals. Being sessile, plants are more vulnerable to drought stress and consequently reduce plant growth and yield. To mitigate the effects of drought stress on plants, it is very crucial to determine the plant response mechanisms against drought stress. Drought response mechanism includes morph-physiological, biochemical, cellular and molecular processes takes place in plants underlying drought stress. These processes include improvement in root system, leaf structure, osmotic adjustment, relative water content and stomata regulation. In addition, calcium and phytohormone (Abscisic acid, Jasmonic acid, Salicylic acid, Auxins, Gibberellins, Ethylene etc.) signaling pathways and scavenging of reactive oxygen species are the key mechanisms to cope with drought stress. Moreover, microorganisms such as bacteria and fungi also have an important role in drought tolerance enhancement. To further elucidate and improve drought tolerance in plants, quantitative trait loci, transgenic approach and application of exogenous substances (nitric oxide, 24-epibrassinoide, glycine betaine and proline) are very crucial. Hereby, the present study integrates various mechanisms of drought tolerance in plants.

Chromium is the second most common metal pollutant in the soil, sediments and groundwater, due to its extensive industrial application, hence posing a serious environmental concern. Various remediation approaches including phytoremediation have been proposed to remediate chromium polluted waters and soils. In the present research, a total of sixty-one plant species belongs to thirty families were analyzed for the concentration of Chromium. Chromium was analyzed in the soil of the root zone, root and shoot of each plant. The concentration of chromium in the soil of different sites and plant parts (roots and shoots) was found in mg/kg in the range of 0.33–48.73, 8–1233.3 and 10.23–568.33 respectively. The highest concentration of chromium was found in the soil of site Site 41 (48.73 mg/kg) followed by Site 18 (47.83 mg/kg) and Site 6 (45.33 mg/kg). Among the analyzed plants, the highest concentration of chromium in mg/kg was found in the root of Cannabis sativa (1233.3) while its highest concentration was found in the shoot of Allium griffithianum (568.33). Phytoremediation potential of the analyzed plants was evaluated by the calculation of Bioconcentration Factor (BCF), Translocation Factor (TF) and Biological Accumulation Coefficient (BAC). Thirty-eight plant species showed feasibility for the phytostabilization of chromium (Cr_Excluders) based on BCF value and the concentration of chromium in the root. Plants i.e. Argyrolobium stenophyllum, Silybum marianum, Bryophyllum daigremontianum, Limonium macrorhabdon, Calendula arvensis and Delphinium suave were found the most efficient plant for the phytostabilization of chromium. Fifteen plant species showed feasibility for the phytoextraction of chromium (Cr_Indicators) based on TF value. The most efficient plant's species among them for the phytoextraction of chromium are Rosularia adenotricha, Catharanthus roseus, Allium griffithianum, Himalaiella heteromalla, Stellaria media, Salvia moorcroftiana and Marrubium vulgare. Based on BCFs, TFs and BACs value and the concentration of chromium in plant shoot six plant species Allium griffithianum, Catharanthus roseus, Himalaiella heteromalla, Geranium rotundifolium, Marrubium vulgare and Solanum nigrum were found chromium hyperaccumulators.

Salvia santolinifolia is a medicinal plant and an efficient in vitro conservation system is established. The influence of N6Benzylaminopurine (BAP), N6-(2-isopentyl)-adenine (2iP) and Kinetin at various concentrations were evaluated, single and in mixture with NAA (Naphthalene acetic acid) for the production of auxiliary shoots from nodal explants of S. santolinifolia. BAP at 3.0 mg/L in MS1 media produced maximum (11.66±3.38) number of shoots while elongated (5.37±1.45) shoots were produced in the MS2 medium in subcultures at 2.0 mg/L of 2iP. Least number of shoots were formed when auxin and cytokinin were used in combination. Length of culture, age was an important consideration for the initiation and development of roots. Rooting of shoot was attained with Indol-3-butyric acid (IBA) (3.0 mg/L) from shoots of 4th, 5th and 6th subculture while shoots taken from 1st, 2nd and 3rd subculture failed to form roots.

Nickel is a known hepatotoxic, haemotoxic, pulmonary toxic, nephrotoxic, reproductive toxic, carcinogenic, phytotoxic and neurotoxic agent. The adverse ecological impacts from unnecessary heavy metals include contamination of water and soil which pose serious threats to human health. This study was conducted to screen plants for the phytoremediation of nickel from sixty-one sites of the Lower Dir. Nickel-metal was analyzed in the soil, roots and shoots of plants. The total concentration of nickel in soil, roots and shoots was found to be in the range of 1.03-18.98, 12.63-540.73 and 12.00-295.86 mg kg–1 dry weight basis (DW) respectively. The highest nickel contents were present in the roots of Xanthium strumarium (540.73) and shoots of Bryophyllum daigremontianum (295.86). None of the plant species were identified as hyper accumulators for nickel but based on BCF, TF and BAC values most of the species showed feasibility for its phytoextraction and phytostabilization. Xanthium strumarium, Filago hurdwarica, Ranunculus arvensis, Medicago lupulina, Cannabis sativa, Geranium rotundifolium and Cerastium glomeratum are suggested for the phytostabilization of nickel whereas Bryophyllum daigremontianum, Rosularia adenotricha, Iris germanica, Asplenium dalhousiae and Isatis tinctoria for the phytoextraction of soil contaminated with nickel.

The present research aimed to determine the lowest levels of three heavy metals (Pb, Cd and Cu) to which the larvae of Southern House Mosquito, Culex quinquefasciatus are susceptible in water. The ...

Background: Germin-like Proteins (GLPs) play an important role in various stresses. Rice contains 43 GLPs, among which many remain functionally unexplored. The computational analysis will provide significant insight into their function. Objective: To find various structural properties, functional importance, phylogeny and expression pattern of all OsGLPs using various bioinformatics tools. Methods: Physiochemical properties, sub-cellular localization, domain composition, Nglycosylation and Phosphorylation sites, and 3D structural models of the OsGLPs were predicted using various bioinformatics tools. Functional analysis was carried out with the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) and Blast2GO servers. The expression profile of the OsGLPs was predicted by retrieving the data for expression values from tissuespecific and hormonal stressed array libraries of RiceXPro. Their phylogenetic relationship was computed using Molecular and Evolutionary Genetic Analysis (MEGA6) tool. Results: Most of the OsGLPs are stable in the cellular environment with a prominent expression in the extracellular region (57%) and plasma membrane (33%). Besides, 3 basic cupin domains, 7 more were reported, among which NTTNKVGSNVTLINV, FLLAALLALASWQAI, and MASSSF were common to 99% of the sequences, related to bacterial pathogenicity, peroxidase activity, and peptide signal activity, respectively. Structurally, OsGLPs are similar but functionally they are diverse with novel enzymatic activities of oxalate decarboxylase, lyase, peroxidase, and oxidoreductase. Expression analysis revealed prominent activities in the root, endosperm, and leaves. OsGLPs were strongly expressed by abscisic acid, auxin, gibberellin, cytokinin, and brassinosteroid. Phylogenetically they showed polyphyletic origin with a narrow genetic background of 0.05%. OsGLPs of chromosome 3, 8, and 12 are functionally more important due to their defensive role against various stresses through co-expression strategy. Conclusion: The analysis will help to utilize OsGLPs in future food programs.

Transcriptional factors are the major regulators of plant signaling pathways in response to environmental stresses i.e., drought, salinity and cold. Hereby, the GhMYB108-like was characterized to determine whether it regulate these stresses. The GhMYB108-like cDNA consisted of 1107 base pairs (bp) with 807 open reading frame encoded a protein of 268 amino acids. Its isoelectric point and molecular weight are 5.51 and 30.3 kDa respectively. Phylogenetic analysis and online databases revealed that GhMYB108-like proteins are closely related with the Arabidopsis thaliana MYB2. Important cis-elements were detected in the promotor region of GhMYB108-like responding to stresses and phytohormones. The 3D structure of GhMYB108-like protein has been predicted. In addition, various physico-chemical properties of GhMYB108-like have been determined. Subcellular localization confirmed that GhMYB108-like are nuclear localized protein. Quantitative expression analysis showed that polyethylene glycol and salt treatments significantly induced the expression of GhMYB108-like. Overall, our findings suggest that GhMYB108-like is an important gene that would plays important regulatory role in response to drought and salt stresses.

This review presents an overview of published studies for a better understanding of the anti-mutagenic potential of medicinal plants and the precise indications for the utilization of natural compounds as chemo-preventive agents. Reports on the anti-mutagenic potential of medicinal plants published from 1997 to 2019 were searched through different scientific databases using the following keywords: medicinal plants and mutagens, carcinogens, the anti-mutagenic potential of medicinal plants. The data relevant to the anti-mutagenic potential of some common medicinal plants is summarized in this mini-review. These medicinal plants include Carum carvi, Withania somnifera, Panax ginseng, Mentha spicata, Curcuma zedoaria, Cassia angustifolia, Cymbopogon citrates, Ipomoea batatas, Glycyrrhiza glabra, Citrullus colocynthis, Capsicum annuum and Asparagus racemosus. An overview of the identified molecules or enzymes being targeted is also presented, with a focus on anti-carcinogenic and/or anti-mutagenic activity. The recent advancements in the research on medicinal plants pave the way for the better understanding and future prospects of the use of natural components as chemo-preventive and chemotherapeutic agents.

Herein, we report an eco-friendly, facile, one-pot, green synthesis of nanoceria for multiple biomedical applications. In the study, cerium oxide nanoparticles (CeO2-NPs) were synthesized using a simple aqueous extract of Aquilegia pubiflora as an effective reducing and capping agent. The biosynthesized nanoparticles were characterized via UV-vis spectroscopy, X-ray powder diffraction (XRD), high-performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and Raman spectroscopy. The NPs were highly stable, exhibited high purity, and had a spherical morphology and mean size of 28 nm. FTIR and HPLC studies confirmed the successful capping of bioactive compounds on the nanoparticles. The well-characterized NPs were evaluated for a number of biomedical applications, and their antimicrobial (antifungal, antibacterial, and antileishmanial), protein kinase inhibition, anticancer, antioxidant, anti-diabetic and biocompatibility properties were studied. Our results showed that the Aquilegia pubiflora mediated CeO2-NPs were highly active against fungal strains, compared to the tested bacterial strains, with Aspergillus niger resulting in the largest zone of inhibition (15.1 ± 0.27 mm). The particles also exhibited dose dependent leishmanicidal activity with significant LC50 values toward both the amastigote (114 μg mL−1) and promastigote (97 μg mL−1) forms of the parasite Leishmania tropica (KWH23). The NPs were found to be moderately active against the HepG2 cell line, showing 26.78% ± 1.16% inhibition at 200 μg mL−1. Last but not least, their highly biocompatible nature was observed with respect to freshly isolated human red blood cells (hRBCs), making the greenly synthesized CeO2-NPs a novel candidates for multidimensional medical applications.

The current study was carried out to explore the phytochemical, antioxidant potential and antibacterial activities of the crude methanolic extract of A. santolinifolia Turcz. Ex Besser. The antioxidant activity was carried out by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assay, while methanolic extract displayed the highest scavenging activity (DPPH) was 61.31µg/ml on Artemisia santolinifolia root and the lowest (51.05µg/ml) was record for their leaves. Similarly, in (ABTS) the highest activity (89.16µg/ml) was recorded for roots of A. santolinifolia followed by leaves (68.14µg/ml). In low inhibitory concentration assay, the crude methanolic extracts showed significant inhibition against all tested microbes on different concentrations like 25 µg/ml, 50 µg/ml, and 100 µg/ml. The leaves extract of A. santolinifolia AsL showed MIC of 12.5µg/ml for B. subtilis, a gram-positive bacterium, 50µg/ml for gram positive bacteria S. aureus and 37.5 µg/ml for gram negative bacteria P. aeruginosa that is almost equal to the response of standard ciprofloxacin. Our current study revealed that Artemisia santolinifolia root (AsR) exhibited a significant antioxidant potential while AsL showed good antibacterial effect which is suggested to be used for treatment and management of different infectious diseases.

The study was aimed to evaluate the therapeutic effects of Zizyphus oxyphyla leaves methanolic (ZOX-LME), on serum liver, kidney and hematology along with other serum parameters in Carbon tetrachloride (CCl4) intoxicated rabbits. Experimental animals were divided into five groups, six rabbits in each. These were: group NC (normal control), group, TC (toxic control) and group ST i.e. silymarine administered group at dose rate (50) mg/kg body weight (BW). Group ET1 and group ET2 treated with (ZOX-LME) at dose 200 mg/kg BW and 400 mg/kg BW. CCl4 administration caused significant (P> 0.05) impairment in serum liver enzymes, blood factors and other serum indices. Treatment with (ZOX-LME) significantly (P 0,05) nas enzimas hepáticas séricas, fatores sanguíneos e outros índices séricos. O tratamento com (ZOX-LME) reduziu significativamente (P < 0,05) e normalizou os níveis de alanina transaminase (ALT), aspartato transaminase (AST) e fosfatase alcalina (ALP) e os índices hematológicos. Também foi observada redução significativa (P < 0,05) nas concentrações de creatinina, ureia, ácido úrico, nitrogênio ureico no sangue (BUN), albumina e glicose. Os níveis alterados de perfil lipídico e eletrólitos séricos (Ca, Mg, Cl, Na, K e P) foram significativamente (P < 0,05) mudando em direção aos níveis normais com a alimentação (ZOX-LME). Além disso, a ingestão de (ZOX-LME) causou melhora significativa nos níveis de GSH, GST e CAT, enquanto reduzia os níveis de TBARS, exibindo capacidade antioxidante. Também (ZOX-LME) mostrou inibição aumentada contra a eliminação percentual do radical livre 2, 2-difenila-1-picrilehidrazila (DPPH). Efeitos de normalização significativos (P < 0,05) foram observados com altas doses de 400 mg / kg de peso corporal de (ZOX-LME) e foram equivalentes aos grupos administrados com silimarina. O estudo histológico do fígado confirmou a atividade hepatoprotetora e curativa renal de (ZOX-LME).

Recently, genome‐editing technology like clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 has improved the translational gap in the treatments mediated through gene therapy. The advantages of the CRISPR system, such as, work in the living cells and tissues, candidate this technique for the employing in experiments and the therapy of central nervous system diseases. Parkinson's disease (PD) is a widespread, disabling, neurodegenerative disease induced by dopaminergic neuron loss and linked to progressive motor impairment. Pathophysiological basis knowledge of PD has modified the PD classification model and expresses in the sporadic and familial types. Analyses of the earliest genetic linkage have shown in PD the inclusion of synuclein alpha (SNCA) genomic duplication and SNCA mutations in the familial types of PD pathogenesis. This review analyzes the structure, development, and function in genome editing regulated through the CRISPR/Cas9. Also, it explains the genes associated with PD pathogenesis and the appropriate modifications to favor PD. This study follows the direction by understanding the PD linking analyses in which the CRISPR technique is applied. Finally, this study explains the limitations and future trends of CRISPR service in relation to the genome‐editing process in PD patients' induced pluripotent stem cells., Parkinson's disease is a widespread, disabling, neurodegenerative disease induced by dopaminergic neuron loss and is linked to progressive motor impairment. Recently, genome‐editing technology like clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 has improved the translational gap in the treatments mediated through gene therapy. The advantages of the CRISPR system, such as, work in the living cells and tissues.

BackgroundCurcuma longa(Turmeric) is a traditionally used herb in wound healing. The efficacy of fresh turmeric paste to heal wounds has already been investigated in multiple ethnobotanical studies. Wnt/β-catenin signaling pathway plays a significant role in wound healing and injury repair processes which has been evident in different in vitro studies. This study aims to analyze the potentiality of curcuminoids (curcumin I, curcumin II and curcumin III) fromCurcuma longato bind and enhance the activity of two intracellular signaling proteins- casein kinase-1 (CK1) and glycogen synthase kinase-3β (GSK3B) involved in Wnt/β-catenin signaling pathway. This study is largely based on a computer-based molecular docking program which mimics the in vivo condition and works on a specific algorithm to interpret the binding affinity and poses of a ligand molecule to a receptor. Subsequently, drug likeness property, ADME/Toxicity profile, pharmacological activity, and site of metabolism of the curcuminoids were also analyzed.ResultsCurcumin I showed better affinity of binding with CK1 (− 10.31 Kcal/mol binding energy) and curcumin II showed better binding affinity (− 7.55 Kcal/mol binding energy) for GSK3B. All of the ligand molecules showed quite similar pharmacological properties.ConclusionCurcumin has anti-oxidant, anti-carcinogenic, anti-mutagenic, anti-coagulant, and anti-infective properties. Curcumin has also anti-inflammatory and wound healing properties. It hastens wound healing by acting on different stages of the natural wound healing process. In this study, three curcumins fromCurcuma longawere utilized in this experiment in a search for a drug to be used in wound healing and injury repair processes. Hopefully, this study will raise research interest among researchers.

Aurivillius single-phase Bi2.5Nd0.5Nb1.5Fe0.5O9 (BNNF) ceramics was synthesized via solid state mixed oxide sintering method. Electrical properties of the samples were intensely studied in a wide range of temperature (25 °C–650 °C). Dielectric behaviors were investigated by dielectric and impedance spectroscopies in the range (40 Hz−10 MHz) of frequency. Single dielectric relaxation was observed due to the grains contribution. The phase transition temperature (TC) was noticed at ∼270 °C. Kinetic analysis of dielectric data was used to understand the possible relaxation-conduction behaviors in the ceramics. Above ∼250 °C, relaxation and conduction of the ceramics was assigned to the motion of ionized oxygen vacancies. The Aurivillius ceramics has promising industrial applications in dielectric capacitors and piezoelectric transducers due to low leakage below ∼200 °C. The results could be valuable for designing and/or modifying properties of BNNF related materials.

Aerolysin (aer) is one of the most important and abundant virulence factors in the infection of fish by Aeromonas veronii. A comprehensive study on the molecular characterization and pathogenicity of the aer gene from 34 A. veronii isolates from diseased carp and catfish was carried out and its interactome was analysed to observe the functional correlations between aer and other proteins within the A. veronii network. The PCR-based amplification of aer from the 34 isolates of A. veronii showed more aer-positive isolates from catfish with a high pathogenic potential in the in vivo challenge test than the carp fish. The analysis of aer gene sequence from challenged fish revealed significant sequence divergence according to the types and geographical distribution of the fish. The networking analysis of aer from the model A. veronii B565 revealed histidine kinase (cheA) as the most functional interacting partner. The study of the interaction between aer from the experimental A. veronii and cheA demonstrated that the A chain of cheA plays a more important role than the corresponding B chain during contact, and a linker sequence of 15 residues controlled the entire interaction process. Therefore, cheA could be an excellent drug target for controlling A. veronii infection of fish.

The purpose of this study was to evaluate the allelopathic effect of weeds (Avena fatua, Melilotus officinalis and Polypogon hissaricus) on germination, growth, dry biomass and chlorophyll concentration of three cultivars of wheat (Ata Habib, Pirsabaq and Serin). In germination test, different concentrations of aqueous extracts (5, 10 and 15 g/l) of the three weeds significantly reduced percent germination; however, 15 g/l extract of M. officinalis resulted in complete failure of germination of cultivar Pirsabaq. In pot culture, root and shoot length, chlorophyll concentration and seedling dry biomass of the three wheat varieties showed differential responses to different weeds. Aqueous extract at 15 g/l of A. fatua increased root and shoot length and dry biomass of cultivar Pirsabaq; however, these parameters were significantly retarded in other two wheat cultivars by extract of weeds. Moisture content of the cultivars did not show any response to allelopathic stress of the weeds. In contrast, chlorophyll concentration in Pirsabaq and Serin was significantly increased by aqueous extract of all the weeds but reduced it in cultivar Ata Habib by 50%. In general, Ata Habib was found to be the most sensitive cultivar to the imposed allelopathic stress. The phytotoxic potential of three weeds was found in the order of A. fatua > M. officinalis > P. hissaricus.

The western Himalayan region in northern Pakistan is one of the most sensitive hotspots to climate change, due to the rapidly increasing population and delicate mountainous ecosystem. The relatively limited observed instrumental record impedes our understanding of long-term climate variability and their assessment. Using standard dendrochronological techniques, a 395-year (1620 to 2014 C.E.) tree-ring chronology of Abies pindrow (Royle) (Himalayan fir) was developed from the western Himalayan region in northern Pakistan. The results of the growth-climate relationship demonstrated that the radial growth of Abies pindrow was limited by minimum temperature. Using a robust reconstruction model, a 310-year (1705 to 2014 C.E.) minimum temperature was reconstructed from the western Himalayan region in northern Pakistan. The reconstructed minimum temperature accounts for 38% variance of the actual minimum temperature, and provides the evidences of Dalton minimum and modern maximum periods. The coldest years in the reconstruction were 1726, 1727, 1892, 1921, and 2001, whereas the hottest years were 1789, 1807, 1814, 1846, 2011, and 2013. Multi-taper method (MTM) spectral analysis showed a significant shorter quasi-cycles (2.3 to 3.5 years) and decadal cycles (11.5 to 17.5 years), suggesting a possible teleconnections with El Nino Southern Oscillation (ENSO), and Northern Atlantic Oscillation (NAO) and Atlantic Multidecadal Oscillation (AMO) respectively.

The New Delhi Metallo-β-lactamase (NDM) is among the most threatening forms of carbapenemases produced by K. pneumoniae, well-known to cause severe worldwide infections. The molecular epidemiology of blaNDM-1-harboring K. pneumoniae is not well elucidated in Pakistan. Herein, we aim to determine the antibiotics-resistance profile, genes type, molecular type, and plasmid analysis of 125 clinically isolated K. pneumoniae strains from urine samples during July 2018 to January 2019 in Pakistan. A total of 34 (27.2%) K. pneumoniae isolates were carbapenemases producers, and 23 (18.4%) harbored the blaNDM-1 gene. The other carbapenemases encoding genes, i.e., blaIMP-1 (7.2%), blaVIM-1 (3.2%), and blaOXA-48 (2.4%) were also detected. The Multi Locus Sequence Typing (MLST) results revealed that all blaNDM-1-harboring isolates were ST11. The other sequence types detected were ST1, ST37, and ST105. The cluster analysis of Xbal Pulsed Field Gel Electrophoresis (PFGE) revealed variation amongst the clusters of the identical sequence type isolates. The blaNDM-1 gene in all of the isolates was located on a 45-kb IncX3 plasmid, successfully transconjugated. For the first time, blaNDM-1-bearing IncX3 plasmids were identified from Pakistan, and this might be a new primary vehicle for disseminating blaNDM-1 in Enterobacteriaceae as it has a high rate of transferability.

Genetic diversity can be investigated through different methods such as morphological traits, biochemical (SDS PAGE) and molecular markers. The present work was conducted on the basis of morphological and biochemical characterization in order to estimate the genetic diversity among the barley landraces; to explore significant variation which can be used in breeding programs. For this purpose, 40 barley landraces were collected from Dir Lower and Swat, districts of KP, Pakistan. A total of 18 traits were recorded, 10 qualitative characters including spike density, awn barbs, glume awn, lemma type and awn color, length of the rechilla hairs, lemma color, auricle pigmentation, photoperiod sensitivity and stem pigmentation. Variation was observed in these qualitative traits through frequency distribution. In quantitative traits maximum variation was found for plant height ranging from 36cm to 50cm, days to flowering range from 130 to 133 days), while 100 seed weight ranged from 36 to 86 g and plant biomass range from 50 to 199.5 g per plant. SDS-PAGE analyses of total seed storage protein resulted in a total of 18 polymorphic bands. Total genetic diversity on the basis of total seed storage protein analysis was 17.5%. In Band, the 14 total genetic diversity was (0.60%) followed by Band 16 (0.58%) and Band 17 (0.55%). Similarly, Band 3 showed diversity, while Band 8, 4 and 5 indicated 0.45, 0.38 and 0.35%, respectively. A cluster dendrogram tree was constructed which was divided into two linkage and further divided into seven clusters. The variation was found among the clusters. The information obtained from the study is useful in planning further crop improvement.