EMvelop stimulation: minimally invasive deep brain stimulation using temporally interfering electromagnetic waves.

Objective.Recently, the temporal interference stimulation (TIS) technique for focal noninvasive deep brain stimulation (DBS) was reported. However, subsequent computational modeling studies on the human brain have shown that while TIS achieves higher focality of electric fields than state-of-the-art methods, further work is needed to improve the stimulation strength. Here, we investigate the idea of EMvelop stimulation, a minimally invasive DBS setup using temporally interfering gigahertz (GHz) electromagnetic (EM) waves. At GHz frequencies, we can create antenna arrays at the scale of a few centimeters or less that can be endocranially implanted to enable longitudinal stimulation and circumvent signal attenuation due to the scalp and skull. Furthermore, owing to the small wavelength of GHz EM waves, we can optimize both amplitudes and phases of the EM waves to achieve high intensity and focal stimulation at targeted regions within the safety limit for exposure to EM waves.Approach.We develop a simulation framework investigating the propagation of GHz EM waves generated by line current antenna elements and the corresponding heat generated in the brain tissue. We propose two optimization flows to identify antenna current amplitudes and phases for either maximal intensity or maximal focality transmission of the interfering electric fields with EM waves safety constraint.Main results.A representative result of our study is that with two endocranially implanted arrays of size4.2 cm×4.7 cmeach, we can achieve an intensity of 12 V m-1with a focality of3.6 cmat a target deep in the brain tissue.Significance.In this proof-of-principle study, we show that the idea of EMvelop stimulation merits further investigation as it can be a minimally invasive way of stimulating deep brain targets and offers benefits not shared by prior methodologies of electrical or magnetic stimulation.

Bacillus licheniformis (MN900686) Mediated Synthesis, Characterization and Antimicrobial Potential of Silver Nanoparticles.

The use of bacteria in the synthesis of silver nanoparticles (AgNPs) emerges as an ecofriendly and exciting approach. In the present study, we reported the biosynthesis of AgNPs by using culture supernatant of the bacteria Bacillus licheniformis (MN900686). The biogenically synthesized AgNPs were confirmed by the change in the color of the culture filtrate from yellow to brown after the addition of AgNO3. Further characterization performed by means of UV vis-spectroscopy showed absorption peak at 414 nm which confirmed the formation of AgNPs. Fourier Transfer infrared (FTIR) confirmed the involvement of biological molecules in the formation of nanoparticles (NPs). The SEM revealed that the NPs have approximately 38 nm size. The agar well diffusion assay was used to determine antibacterial activity while tube dilution method was used to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The human pathogenic bacterial strains i.e., P. aeruginosa (MN900691) and B. subtilis (MN900684), were used as test strains. The anti-bacterial assay against test strains revealed that these NPs showed concentration dependent increased zone of inhibition (ZOI). The maximum ZOI at 25 µL of AgNPs was 20 mm against B. subtilis after 24 hours of incubation. One-way ANOVA test showed significant ZOI (p ≤ 0.05) against B. subtilis. The MIC was ranged from 4.3-6.6 µg/mL while MBC ranged from 8.3 to 6.6 µg/mL. Overall, this study suggested that the biogenically synthesized NPs are an effective alternative source of antimicrobials against pathogenic bacteria.

A Comparative Analysis of Antimicrobial, Antibiofilm and Antioxidant Activity of Silver Nanoparticles Synthesized from Erythrina Suberosa Roxb. and Ceiba Pentandra.

Biofilm forming bacteria can cause serious health problems that are difficult to combat. Silver nanoparticles (Ag-NPs) synthesized from plant extracts have potential to fight and eradicate biofilmforming bacteria. In the present research, AgNPs were synthesized using leaf and bark extract of Erythrina suberosa Roxb. and Ceiba pentandra L. and their antibiofilm, antioxidant and antibacterial activity was checked. Phytochemical analysis of the plant extracts showed important bioactive compounds such as tannins, saponins, steroids, phenolics, alkaloids, flavonoids and glycosides. The AgNPs were synthesized and confirmed by visual color observation and UV-Vis spectrophotometer. Visual color observation showed that the color of the leaf and bark extracts of E. suberosa and C. pentandra turned into brown. UV-Vis spectra analysis showed absorbance peak range between 430-450 nm. The antioxidant activity of the AgNPs was determined by FRAC (Ferrous reducing antioxidant capacity) assay. Synthesized AgNPs from all sources showed significant antioxidant activity. However, antioxidant activity of E. suberosa AgNPs was significant compared to other sources. Antibacterial activity and biofilm forming assay was analyzed against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The synthesized AgNPs silver nanoparticles showed significant (p ≤ 0.05) antibacterial activity against all the bacteria. The maximum zone of inhibition was found in case of E. suberosa AgNPs bark extract against P. aeruginosa was 20±1.154 mm. The results of biofilm forming assay showed that the AgNPs from all sources significantly (p ≤ 0.05) inhibited the activity of biofilms by all the tested bacteria. From results, it can be concluded that AgNPs synthesized from both plants can be used in developing antimicrobial compounds.

Purification and Characterization of Keratinase from Bacillus licheniformis dcs1 for Poultry Waste Processing.

Feather wastes-byproduct of commercial poultry processing plant is produced in large amounts. Keratinolytic enzymes produced by feather degrading bacteria can easily degrade these waste products releasing pure keratin as a residue. The aim of present study was to isolate, and characterize feather degrading bacteria as well as assess the keratinolytic potential of purified enzyme. Three feather degrading bacteria (dps3, wps1 and dcs1) were isolated from feathers of domestic chickens. Preliminary characterization of isolated bacteria revealed these isolates belonging to genus Bacillus. 16S rRNA gene sequencing identified the isolates as B. subtilis dps3 (MW255302), B. cereus wps1 (MW255303) and B. licheniformis dcs1 (MW255304). Cell free supernatant of B. licheniformis dcs1 degraded feathers completely in 14 days indicating its keratinolytic ability. Purification of keratinase enzyme from B. licheniformis dcs1 was performed using column chromatography. SDS-PAGE indicated its molecular weight as 32 KDa. Kerotinolytice activity was maximum at optimum pH of 7 and 45℃ temperature. Enzyme showed the potential to degrade keratin material such as hairs and nails of humans. Findings of current study suggested that purified enzyme possess potential to upgrade nutritional quality of poultry waste containing keratin and might play as important biotechnological tool for keratin hydrolysis.

Correction: Saeed, A., et al. Incidence of Vancomycin-Resistant Phenotype of the Methicillin-Resistant Staphylococcus aureus Isolated from a Tertiary Care Hospital in Lahore. Antibiotics 2020, 9, 3.

Staphylococcus aureus (S. aureus)-associated infections are one of the major threats to public health. The aim of the present study was to determine the antibiotic resistance pattern, as well as the genetic characterization, of methicillin and vancomycin-resistant S. aureus (VRSA) isolated from a tertiary care hospital in Lahore. The S. aureus isolates were isolated from different clinical samples, identified by biochemical testing, and subjected to antibiotic susceptibility testing via the disc diffusion method or broth microdilution method. The methicillin resistance gene (mecA) and vancomycin resistance gene (vanA) were amplified by the polymerase chain reaction. The S. aureus isolates showed high incidences of resistance against methicillin (76%) and moderate incidences of resistance to vancomycin (14%). Isolates were also resistant to several other drugs, such as cefoxitin (76%), ertapenem (83%), ampicillin (81%), tobramycin (78%), moxifloxacin (76%), and tetracycline (74%). An encouraging finding was that 98% of isolates were susceptible to tigecycline, indicating its possible role in the treatment of methicillin-resistant Staphylococcus aureus (MRSA) and VRSA, as well as the multi-drug resistant S. aureus. The mecA gene was detected in 33.3% of tested isolates (10/30), while the vanA gene was also detected in 30% (9/30) of the tested isolates. In conclusion, the frequent presence of methicillin and vancomycin resistance in S. aureus appraises the cautious use of these antibiotics in clinical practices. Furthermore, it is suggested that there should be continuous monitoring of tigecycline treatments in clinical setups in order to delay the development of resistance against it.

Incidence of Vancomycin Resistant Phenotype of the Methicillin Resistant Staphylococcus aureus Isolated from a Tertiary Care Hospital in Lahore.

Staphylococcus aureus (S. aureus) associated infections are one of the major threats to public health. The aim of the present study was to determine the antibiotic resistance pattern as well as the genetic characterization of methicillin and vancomycin resistant S. aureus (VRSA) isolated from a tertiary care hospital in Lahore. The S. aureus were isolated from different clinical samples, identified by biochemical testing, and subjected to antibiotic susceptibility testing by disc diffusion method or broth microdilution method. Methicillin resistance gene (mecA) and vancomycin resistance gene (vanA) were amplified by polymerase chain reaction. The S. aureus isolates showed high incidence of resistance against methicillin (76%) and moderate incidence of resistance to vancomycin (44%). Isolates were also resistant to several other drugs such as cefoxitin (76%), ertapenem (83%), ampicillin (81%), tobramycin (78%), moxifloxacin (76%), and tetracycline (74%). An encouraging finding was that 98% of isolates were susceptible to tigecycline, indicating its possible role in the treatment of MRSA, VRSA, as well as multi-drug resistant S. aureus. The mecA gene was detected from 33.3% (10/30) while vanA gene was detected from 46.6% (14/30) of the tested isolates. In conclusion, frequent presence of methicillin and vancomycin resistance in S. aureus appraises the cautious use of these antibiotics in clinical practices. Furthermore, it is suggested that there should be continuous monitoring of tigecycline treatments in clinical setups in order to delay the development of resistance against it.

Amelioration of allergic asthma by Ziziphora clinopodioides via upregulation of aquaporins and downregulation of IL4 and IL5.

Ziziphora clinopodioides has been frequently used as an anti asthmatic plant in traditional medication. Recent work explores the anti-asthmatic activity of Z. clinopodioides in allergen-induced asthmatic mice. Intraperitoneal sensitization followed by intranasal challenge were given with ovalbumin (allergen) to develop allergic asthma. Investigational groups of animals were administered with drug methylprednisolone (MP) (15 mg/kg body weight), n-hexane fraction, ethylacetate fraction, and methanolic extract of Z. clinopodioides extract (500 mg/kg b.w.) for successive 07 days. Hematoxyline and eosin (H&E) and periodic acid-Schiff (PAS) stains were used to evaluate histopathological parameters on lung tissues. As an index of lungs tissues edema, wet/dry weight ratio of lungs was determined. Evaluation of expression levels of AQP1, AQP5, IL4, and IL5 was conducted by using RT-PCR. The data exhibited that both Z. clinopodioides and MP attenuated differential and total leukocyte counts in hematological examination i.e. in BALF and blood. Treatment with Z. clinopodioides also caused suppression of inflammatory cell infiltration and expression levels of IL4 and IL5, the later could have caused attenuation of pulmonary inflammation. The study also found decline in lung wet/dry ratio and goblet cellh hyperplasia in treated groups which indicates amelioration of lung edema. Treatment with Z. clinopodioides significantly increased the expression levels of aquaporin-1 and -5, which could have led to reduction in lung edema. The treatment with MP showed comparable results to Z. clinopodioides. Current investigation revealed that Z. clinopodioides possessed anti-asthmatic property which might be accredited to upregulagted AQP1 and AQP5 levels and downregulated IL4 and IL5 levels.

Cyclosporine H Overcomes Innate Immune Restrictions to Improve Lentiviral Transduction and Gene Editing In Human Hematopoietic Stem Cells.

Innate immune factors may restrict hematopoietic stem cell (HSC) genetic engineering and contribute to broad individual variability in gene therapy outcomes. Here, we show that HSCs harbor an early, constitutively active innate immune block to lentiviral transduction that can be efficiently overcome by cyclosporine H (CsH). CsH potently enhances gene transfer and editing in human long-term repopulating HSCs by inhibiting interferon-induced transmembrane protein 3 (IFITM3), which potently restricts VSV glycoprotein-mediated vector entry. Importantly, individual variability in endogenous IFITM3 levels correlated with permissiveness of HSCs to lentiviral transduction, suggesting that CsH treatment will be useful for improving ex vivo gene therapy and standardizing HSC transduction across patients. Overall, our work unravels the involvement of innate pathogen recognition molecules in immune blocks to gene correction in primary human HSCs and highlights how these roadblocks can be overcome to develop innovative cell and gene therapies.

Preventing the N-terminal processing of human interferon α-2b and its chimeric derivatives expressed in Escherichia coli.

We have previously shown that human interferon α-2b (IFN) produced in Escherichia coli (E. coli) is heterogeneous at the N-terminal, with three major species (Ahsan et al., 2014). These are: (a) the direct translation product of the gene retaining the N-terminal methionine, (b) a species from which the methionyl residue has been removed by E. coli methionyl aminopeptidase to give the native interferon α-2b and (c) in which the N-terminal Cys residue of the latter contains an acetyl group. In this paper we overcome this heterogeneity, using engineered interferon derivatives with phenylalanine residue directly downstream of the N-terminal methionine (Met-Phe-IFN). This modification not only prevented the removal of the N-terminal methionine by E. coli methionyl aminopeptidase but also the subsequent N-acetylation. Critically, Met-Phe-IFN had enhanced activity in a biological assay. N-terminal stabilization was also achieved by fusing human cytochrome b5 at the N-terminal of interferon (b5-IFN-chimera). In this case also, the protein was more active than a reciprocal chimera with cytochrome b5 at the C-terminal of interferon (Met-IFN-b5-chimera). This latter protein also had a heterogeneous N-terminal but addition of phenylalanine following Met, (Met-Phe-IFN-b5-chimera), resolved this problem and gave enhanced biological activity.

Characterization and bioassay of post-translationally modified interferon α-2b expressed in Escherichia coli.

Examples of N-terminal acetylation are rare in prokaryotic systems, but in this study, we report one such example in which N-terminal Cys residue of recombinant human interferon α-2b produced in Escherichia coli is a favourite site for N(α)-acetylation. The recombinant protein following Q-sepharose chromatography gave a single band on PAGE analysis. However, on reverse phase HPLC the material separated into three peaks. These were characterized by mass spectrometric techniques as: (a) the direct translation product of the gene retaining the N-terminal methionine, (b) a species from which the methionyl residue had been removed by E. coli methionyl aminopeptidase to give the native interferon α-2b and (c) in which the N-terminal Cys residue of the latter contained an acetyl group. Tryptic digestion of interferon α-2b gave fragments linking Cys(1) to Cys(98) and Cys(29) to Cys(138), while that of N(α)-acetyl-interferon α-2b gave the Cys(1)-Cys(98) fragment with an additional mass of 42 attributed to an acetylated N-terminal. Bioassay of the derivatives showed that N(α)-acetyl-interferon α-2b had 10% of the activity of interferon α-2b. The results suggest that the lower activity derivative seen here in E. coli may also be produced when the protein is produced in yeast.