Scanning electron microscopy of bio‐fabricated Fe2O3 nanoparticles and their application to control brown rot of citrus

Citrus is the leading fruit crop of Pakistan and exported to different parts of the world. Due to suitable weather condition, this crop is affected by different biotic factors which seriously deteriorate its quality and quantity. During the months of November 2018 to January 2019, citrus brown rot symptoms were recurrently observed on sweet oranges in National Agricultural Research Centre (NARC), Islamabad. Causal agent of citrus brown rot was isolated, characterized, and identified as Fusarium oxysporum. For environment‐friendly control of this disease, leaf extract of Azadirachta indica was used for the green synthesis of iron oxide (Fe₂O₃) nanoparticles. These nanoparticles were characterized before their application for disease control. Fourier transform infrared spectroscopy (FTIR) of these synthesized nanoparticles described the presence of stabilizing and reducing compounds like alcohol, phenol, carboxylic acid, and alkaline and aromatic compounds. X‐Ray diffraction (XRD) analysis revealed the crystalline nature and size (24 nm) of these nanoparticles. Energy dispersive X‐Ray (EDX) analysis elaborated the presence of major elements in the samples. Scanning electron microscopy (SEM) confirmed the spinal shaped morphology of prepared nanoparticles. Successfully synthesized nanoparticles were evaluated for their antifungal potential. Different concentrations of Fe₂O₃ nanoparticles were used and maximum mycelial inhibition was observed at 1.0 mg/ml concentration. On the basis of these findings, it could be concluded that Fe₂O₃ nanoparticles, synthesized in the leaf extract of A. indica, can be successfully used for the control of brown rot of sweet oranges.     

一种板栗榛子自动脱蓬分选一体机的结构设计

板栗的营养丰富,市场需求日益渐增,但是问题是在采摘时很难将板栗毛壳去掉。传统脱蓬一般用剪刀或者其他锋利尖锐的工具将板栗毛壳破开取出里面的板栗坚果,劳动强度大,生产效率低。文章针对此类坚果特征,设计一款板栗榛子自动脱蓬分选一体机,该机兼有脱蓬和分选的功能,由电动机驱动,先在脱蓬机构中将板栗、榛子等的带刺外壳挤压破碎并去除,然后传送到分选机构将脱蓬后的板栗或榛子进行分级筛选,便于后续分级别售卖,从而减轻人工脱蓬和分选的劳动强度,提高生产效率。     

Detection of new antibiotic resistance gene profile in <i>Escherichia coli</i> associated with avian leukosis virus infection from broiler chickens

The Escherichia coli (E. coli) is prevailing worldwide, but the epidemiology of E. coli infections feature regional distribution characteristics to some extent. E. coli, as a zoonotic pathogen, can be transferred from animals to humans through food chain or via contact with wounds, causing a public health risk. We reported the swelling of proventriculus and tracheal bleeding following the death in two broiler chickens (Gallus gallus domesticus) from Beijing, China. To investigate whether a virus was involved in the infection, Madin Darby Bovine Kidney (MDCK) cells were co-cultured with supernatants of proventriculus, trachea and spleen homogenates. The avian leucosis virus was detected in the samples of proventriculus and trachea, but the avian influenza virus, the Newcastle disease virus and the avian infectious laryngotracheitis virus were not detected. E. coli isolates were resistant to almost all the antimicrobial as tested except for the combinations of amoxicillin/clavulanic acid and sulfamethoxazole/trimethoprim. PCR tests demonstrated the presence of antibiotic resistance genes in these E. coli isolates and further research revealed a novel gene profile with the presence of CTX-M-1, gyrA, gyrB, oqxA, oqxB, parC and Sul2 antibiotic resistance genes in a strain isolated from a proventriculus sample. These results demonstrated that the presence of antibiotic resistant E. coli would not necessarily cause outbreak of large-scale disease. However, when the bacteria carrying new antibiotic resistance genes enter the environment, it may result in the development of more virulent strains which will potentially impact human and animal health.     

Organogenesis and plant regeneration of <i>Arachis villosa</i> Benth. (Leguminosae) through leaf culture

With the aim of developing an efficient plant regeneration protocol, leaflet explants of three accessions of Arachis villosa Benth. (S2866, S2867 and L97) were cultured on basic Murashige and Skoog medium supplemented with different combinations of plant growth regulators: α-naphthalenacetic acid, indole-3-butyric acid, 6-benzylaminopurine, kinetin and thidiazuron. The accession L97 was the only one able to differentiate buds through indirect organogenesis. The most suitable combination for bud regeneration was the basic medium added with 13.62 μM thidiazuron and 4.44 μM 6-benzylaminopurine. These results show the important role of the genotype in morphogenetic responses and the organogenetic effect of thidiazuron in Arachis villosa accession L97. A thidiazuron lacking media (only 0.54 μM α-naphthalenacetic acid, 13.95 μM kinetin and 13.32 μM 6-benzylaminopurine were added) promoted the elongation of the regenerated buds. Adventitious rooting was achieved 90 days after the isolated shoots were transferred to a rooting medium containing 0.54 μM α-naphthalenacetic acid.     

Phylogenetic analysis and target gene prediction of miR477 gene family in grape

To understand the molecular characteristics of the miR477 gene family of grape (Vvi-miR477) and to predict its target genes, the Vvi-miR477 genes were identified from previous small RNA sequencing data, then phylogenetic analysis and prediction of target gene were conducted. The Vvi-miR477 family consists of two precursor sequences and three mature sequences. The miR477 family members were mostly 19-22nt in length. The sequence is relatively conservative. Vvi-MIR477a and Vvi-MIR477b are located on chromosomes 1 and 2, respectively. These precursor sequences can form the typical stable stem-loop structure. Their minimum folding free energy is −39.10 kcal/mol and −50.90 kcal/mol, respectively. The MIR477 family can be divided into three groups. The prediction of target genes showed that Vvi-miR477 targets 26S proteasome, DEAD-box, GRAS family protein, Protein Phosphatase 2C, etc. The GO function of target genes was mainly enriched to six categories. The catabolic process, carboxylic ester hydrolase activity is shown to be high. This study provided a theoretical basis for further exploration of the molecular mechanism of miR477 in grape berry ripening.     

Identification of ages and determination of paeoniflorin in roots of Paeonia lactiflora Pall. From four producing areas based on growth rings

Growth rings were used to determine the root age of medicinal Paeonia lactiflora from four producing areas, and their corresponding paeoniflorin content were measured based on the identification of ages. Different P. lactiflora root samples of different ages were collected from the four major growing areas in China: Bozhou, Anhui Province; Pan'an, Zhejiang Province; Zhongjiang, Sichuan Province; and Heze, Shandong Province. The relationship between the number of growth rings and age was analyzed using hand sections and paraffin sections. The paeoniflorin content in the roots of different P. lactiflora cultivars from different growing areas was measured using high‐performance liquid chromatography (HPLC). The growth rings in the P. lactiflora roots were consistent with the age of the plant from Heze, Zhongjiang, Pan'an, whereas that for the P. lactiflora from Bozhou was one less than the age of the plant. The HPLC results show that the paeoniflorin content was highest in P. lactiflora ‘Baihuachuanshaoyao,’ followed by ‘Baihuahangshaoyao,’ ‘Honghuachuanshaoyao,’ and ‘Honghuahangshaoyao,’ ‘Bozhoushaoyao’ had the lowest levels of paeoniflorin. With increasing age, the paeoniflorin in the roots of the different P.lactiflora cultivars slowly declined or remained the same. In summary, the age of the roots of P. lactiflora from different growing areas can be determined using growth rings. The paeoniflorin content in the roots of P. lactiflora is correlated with cultivar and it was slowly declined with increasing age. Microsc. Res. Tech. 75:1191–1196, 2012. © 2012 Wiley Periodicals, Inc.     

Molecular typing of methicillin and vancomycin-resistant <i>Staphylococcus aureus</i> isolated from clinical specimens by doublelocus sequence typing (DLST) method

Methicillin-resistant Staphylococcus aureus (MRSA) strains are the essential cause of infections in communities and hospitals. The present study was conducted to determine the molecular typing of MRSA, isolated from hospitalized patients, using the double-locus sequence typing (DLST). In total, 280 S. aureus isolated from clinical specimens by phenotypic (catalase, coagulase, DNase, oxacillin, vancomycin screening agar and antibiotic disk diffusion), and molecular methods (PCR for determining the mecA, vanA and nuc genes). The DLST and sequencing was performed for MRSA containing mecA. Out of 280 specimens, confirmed as Staphylococcus aureus (S. aureus), 123 (43.9%) strains were MRSA. The highest resistance toward the erythromycin (15 μg), followed by ciprofloxacin (5 μg), clindamycin (2 μg), tetracycline (30 μg), gentamicin (10 μg) and rifampicin (5 μg), was 98.3%, 97.5%, 94.3%, 90.2%, 83.7% and 41.4%, respectively. Also, the least resistance (0%) was observed in each of teicoplanin (30 μg), linzolide (30 μg), and vancomycin (30 μg). All (100%) of MRSA strains had the mecA, and none of them have had the vanA. The results of DLST showed that the most common sequence types were BPH 2003 and 0217. The DLST type 18-32 was a significant cluster of MRSA. By sequencing MRSA and comparing the dominant types via the DLST, it is possible to establish the etiology of the disease in a much shorter time, and prevent the complications of the disease. Therefore, the combination of partial sequences of clfB and spa can serve as useful genetic markers for MRSA typing. It concluded that the MRSA in our region was relatively high, but no vancomycin resistance was found. The majority of the MRSA DLST type was 18–32.     

Targeting the “undruggable” cancer driver genes: <i>Ras</i>, <i>myc</i>, and <i>tp53</i>

The term “undruggable” is to describe molecules that are not targetable or at least hard to target pharmacologically. Unfortunately, some targets with potent oncogenic activity fall into this category, and currently little is known about how to solve this problem, which largely hampered drug research on human cancers. Ras, as one of the most common oncogenes, was previously considered “undruggable”, but in recent years, a few small molecules like Sotorasib (AMG-510) have emerged and proved their targeted anti-cancer effects. Further, myc, as one of the most studied oncogenes, and tp53, being the most common tumor suppressor genes, are both considered “undruggable”. Many attempts have been made to target these “undruggable” targets, but little progress has been made yet. This article summarizes the current progress of direct and indirect targeting approaches for ras, myc, two oncogenes, and tp53, a tumor suppressor gene. These are potential therapeutic targets but are considered “undruggable”. We conclude with some emerging research approaches like proteolysis targeting chimeras (PROTACs), cancer vaccines, and artificial intelligence (AI)-based drug discovery, which might provide new cues for cancer intervention. Therefore, this review sets out to clarify the current status of targeted anti-cancer drug research, and the insights gained from this review may be of assistance to learn from experience and find new ideas in developing new chemicals that directly target such “undruggable” molecules.     

Tribological behavior of RH ceramics made from rice husk sliding against stainless steel,alumina, silicon carbide,and silicon nitride

The tribological behavior of rice husk (RH) ceramics, a hard, porous carbon material made from rice husk, sliding against stainless steel, alumina, silicon carbide, and silicon nitride (Si₃N₄) under dry conditions was investigated. High hardness of RH ceramics was obtained from the polymorphic crystallinity of silica. The friction coefficients for RH ceramics disks sliding against Si₃N₄ balls were extremely low (<0.1), irrespective of contact pressure or sliding velocity. Transfer films from RH ceramics formed on Si₃N₄ balls. Wear-mode maps indicated that the wear modes were powder formation under all tested conditions, resulting in low specific wear rates (<5×10⁻⁹ mm²/N).     

Co-ordinated combination of Embden-Meyerhof-Parnas pathway and pentose phosphate pathway in <i>Escherichia coli</i> to promote L-tryptophan production

In this study, phosphoenolpyruvate and erythrose-4-phosphate are efficiently supplied by collaborative design of Embden-Meyerhof-Parnas (EMP) pathway and pentose phosphate (PP) pathway in Escherichia coli, thus increasing the L-tryptophan production. Firstly, the effects of disrupting EMP pathway on L-tryptophan production were studied, and the results indicated that the strain with deletion of phosphofructokinase A (i.e., E. coli JW-5 ΔpfkA) produced 23.4 ± 2.1 g/L of L-tryptophan production. However, deletion of phosphofructokinase A and glucosephosphate isomerase is not conducive to glucose consumption and cell growth, especially deletion of glucosephosphate isomerase. Next, the carbon flux in PP pathway was enhanced by introduction of the desensitized glucose-6-phosphate dehydrogenase (zwf) and 6-phosphogluconate dehydrogenase (gnd) and thus increasing the L-tryptophan production (i.e., 26.5 ± 3.2 g/L vs. 21.7 ± 1.3 g/L) without obviously changing the cell growth (i.e., 0.41 h⁻¹ vs. 0.44 h⁻¹) as compared with the original strain JW-5. Finally, the effects of co-modifying EMP pathway and PP pathway on L-tryptophan production were investigated. It was found that the strain with deletion of phosphofructokinase A as well as introduction of the desensitized zwf and gnd (i.e., E. coli JW-5 zwf243 gnd361 ΔpfkA) produced 31.9 ± 2.7 g/L of L-tryptophan, which was 47.0% higher than that of strain JW-5. In addition, the glucose consumption rate of strain JW-5 zwf243 gnd361 ΔpfkA was obviously increased despite of the bad cell growth as compared with strain JW-5. The results of this study have important reference value for the following application of metabolic engineering to improve aromatic amino acids producing strains.     

Response of <i>MaHMA2</i> gene expression and stress tolerance to zinc stress in mulberry (<i>Morus alba</i> L.)

HMA2 (heavy metal ATPase 2) plays a crucial role in extracellular and intracellular Zn²⁺ transport across biomembranes, maintaining ion homeostasis, and playing an important role in the normal physiological metabolism, growth, and development of plants. In our study, a novel HMA2 gene, named MaHMA2, was isolated and cloned from white mulberry (Morus alba L.). The gene sequence obtained was 1,342 bp long, with an open reading frame of 1,194 bp, encoding a protein of 397 amino acids, with a predicted molecular mass of 42.852 kD and an isoelectric point of 7.53. This protein belonged to the PIB-type ATPase transport protein family. We analyzed the expression of the MaHMA2 gene by quantitative real-time PCR. The results showed that the level of MaHMA2 gene expression decreased to a Zn concentration of 800 mg/kg. Malondialdehyde and proline levels increased and responded to increasing Zn when the MaHMA2 gene was silenced, whereas the activities of peroxidase and superoxide dismutase tended to increase in response to increasing Zn²⁺ ion stress concentrations but were lower in the gene-silenced plants. These findings suggested that the MaHMA2 gene played an active role in the tolerance response of mulberry to Zn stress.     

Alternative optimisation strategies and CAM software for multipass rough turning operations

The development of constrained optimisation analyses and strategies for selecting optimum cutting conditions in multipass rough turning operations based on minimum time per component criterion is outlined and discussed. It is shown that a combination of theoretical economic trends of single and multipass turning as well as numerical search methods are needed to arrive at the optimum solution. Numerical case studies supported the developed solution strategies and demonstrated the economic superiority of multipass strategies over single pass. Alternative approximate multipass optimisation strategies involving equal depth of cut per pass, single pass optimisation strategies and limited search techniques have also been developed and compared with the rigorous optimisation strategies. The approximate strategies have been shown to be useful, preferably for on-line applications such as canned cycles on CNC machine controllers, but recourse to the rigorous multipass strategies should be regarded as the reference for use in assessing alternative approximate strategies or for CAM support usage.Nomenclature d _i depth of cut for theith pass - d _opt optimum depth of cut - d _T total depth of cut to be removed - D _i workpiece diameter before theith pass - D _o,D _m initial and final workpiece diameter (afterm passes) - f _i feed for theith pass - f _max,f _min machine tool maximum and minimum feed - f _opt optimum cutting feed - f _sj, V_sj available feed and speed steps in a conventional machine tool - f _sgl, f_rec optimum and handbook recommended single pass cutting feeds - F _pmax maximum permissible cutting force - L workpiece length of cut - m continuous number of passes - m _H next higher integer number of passes from a givenm - m _HW upper limit to the optimum integer number of passesm _opt - m _L next lower integer number of passes from a givenm - m _LW lower limit to the optimum integer number of passesm _opt - m _o optimum (continuous) number of passes - m _opt optimum integer number of passes - N _a machine tool critical rotational speed whenP _a=P _max - N _max,N _min machine tool maximum and minimum rotational speed - n,n ₁,n ₂,K speed, feed and depth of cut exponents and constant in the extended Taylor's tool-life equation - P _a,P _max machine tool low speed and maximum power constraints - T _i tool-life using the cutting conditions for theith pass - T _L loading and unloading time per component - T _R tool replacement time - T _s tool resetting time per pass - T _T production time per component - T _TDi multi-passT _T equation with workpiece diameter effect - T _TDm, T_TDo multi-passT _T equations with constant diameterD _m andD _o, respectively - T _Topt overall optimum time per component - T _Tsgl optimum time per component for single pass turning - T _{T2re c} handbook recommended time per component - V _i cutting speed for theith pass - V _max,V _min machine tool maximum and minimum cutting speed - V _sgl,V _rec optimum and handbook recommended single pass cutting speeds - V _opt optimum cutting speed - a, E, W empirical constants in theP _a/F _pmax/P _max equations - , , feed, depth and speed exponents inF _pmax andP _max equations