Centella asiatica, propolis, and hinokitiol, as natural antibacterial reagents, were integrated into the poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] (PHBH) polymer to produce antibacterial wound dressings, using electrospinning process. The results showed that the fiber diameters and surface morphology of PHBH composite nanofibers were influenced by the addition of ethanol–centella (EC), methanol–centella (MC), ethanol–propolis (EP), and ethanol–hinokitiol (EH) at various ratios compared to pristine PHBH nanofibers. From FT-IR, the nanofibrous samples with higher contents of natural antibacterial substances showed the peaks of carboxylic acid, aromatic ring, and tropolone carbon ring from centella, propolis, and hinokitiol, respectively. Furthermore, the tensile strength of neat PHBH nanofibers was increased from 8.00 ± 0.71 MPa up to 16.35 ± 1.78 MPa by loading of propolis (EP) 7% into PHBH. X-ray analysis explained that the loading of propolis (EP) was also able to increase the crystallinity in PHBH composite nanofibers from 47.0% to 54.5%. The antibacterial results demonstrated that PHBH composite nanofibers containing natural antibacterial products were potent inhibitors against the growth of Escherichia coli and Staphylococcus aureus, amongst them hinokitiol and propolis proved to be the most effective. Additionally, the release studies displayed that centella and hinokitiol had faster release from PHBH composite nanofibers in comparison to propolis. Electrospinning is a straightforward and efficient method to fabricate nonwoven mats with continuous fibers in the range from micrometer down to nanometer. Nowadays, nanofibers produced by electrospinning have received the considerably high attention to be an excellent candidate for many important applications in medical field such as drug delivery , oral caCentella asiatica, a traditional medicine derived from plants, has been used widely in Southeast Asia, India, and other regions for skin treatments. The major biologically active compounds in centella are triterpenoids, which include asiaticoside, madecassoside, asiatic acid, and madecassic acid  (PHB), the basic homopolymer in Polyhydroxyalkanoates (PHA), which are a family of polyesters produced by bacteria through the fermentation of sugars, lipids, fatty acids, alkanes, alkenes, and alkanoic acids [co-3-hydroxyvalerate) (PHBV)/Curcumin nanofibers showed positive effects on wound healing, cell adhesion, and non-toxicity to L929 mouse fibroblast cell line [Poly unit as the polymer matrix regarding crystallinity of the PHBH composite nanofibers, antibacterial activities, and sustained release behaviors. The advantages of this research are to promote the use of natural products over conventional synthetic antiseptics by studying three variants of natural antibacterial products, namely centella, propolis, and hinokitiol. It is also important to present harmlessness for products made of PHBH and natural antibacterial reagents if they are to be used in medical applications.In this study, centella, propolis, and hinokitiol loaded into PHBH were analyzed by scanning electron microscopy (SEM), Fourier Transform Infrared (FTIR) Spectroscopy, tensile testing machine, X-ray diffraction, antibacterial experiment, and UV-visible spectrophotometer, respectively. From these results, the influences of natural products and solvents to PHBH structure and the properties of the resultant nanofibrous composites will be revealed.The surface morphology, chemical characterization, mechanical properties, crystallinity of PHBH, zone of inhibition, and sustained release behaviors of Mw = 5.37 × 105 g/mol%) was provided by Kaneka Corp., Osaka, Japan. The natural antibacterial reagents, centella asiatica powder, propolis powder, hinokitiol powder were purchased from Dins Natural (pvt) Ltd., Matara, Sri Lanka; Stakich, Inc., Troy, USA; and Wako Pure Chemical Industries, Ltd., Osaka, Japan, respectively. Solvents, used to prepare electrospinning solutions, were 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) purchased from Wako Pure Chemical Industries, Ltd., Japan. Acetone, ethanol 99.5%, and methanol were obtained from Wako Pure Chemical Industries, Ltd., Japan.PHBH containing 5.5 mol% of 3HH (Staphylococcus aureus (NBRC 12732) and Escherichia coli (NBRC 3972) for antibacterial activity tests were purchased from the National Institute of Technology and Evaluation, Biological Resource Center (NBRC), Tokyo, Japan.Centella solutions were prepared using either ethanol or methanol the concentrations of centella to solutions were 15% or 30% (w/v), respectively. The propolis solutions were prepared by dissolving propolis in acetone at 10% (w/v) or ethanol with the concentrations of 10% or 30% (w/v), respectively. Each prepared solution was mixed using a magnetic stirrer for 24 h at room temperature. Afterwards, all solutions were filtered to remove insoluble substances. The hinokitiol solution was prepared by dissolving hinokitiol powder in 99.5% ethanol with a concentration of 30% (w/v), also stirred at room temperature for 24 h without filtration.centella (15EC and 30EC), methanol–centella (15MC and 30MC), acetone–propolis (10AP), and ethanol–propolis (10EP and 30EP) were determined from the dry weight of each natural product by evaporating solutions under vacuum. Ethanol–hinokitiol (30EH) mentioned the same as 30% because of no insoluble substances in the ethanol solution.The concentrations of natural products in ethanol–centella, propolis, hinokitiol are displayed in These percentage concentrations of centella (15EC and 30EC) or methanol–centella (15MC and 30MC) were 1%, 5%, and 10% over the total weight of spinning PHBH solutions. Furthermore, ratios of acetone-propolis (10AP), ethanol–propolis (10EP and 30EP), and ethanol–hinokitiol (EH) to total weight of spinning PHBH solutions were 1%, 5%, and 7%, PHBH was dissolved in HFIP at room temperature for 4 h to make solution 2 wt%. The natural antibacterial solutions were added into PHBH HFIP solutions and stirred continuously for 16–20 h to get homogeneous solutions. The ratios of ethanol–The morphology of electrospun nanofibers was investigated by scanning electron microscopy  at an accelerating voltage of 10 kV with various magnifications. The samples were coated with platinum (Pt) in a sputtering device for 60 s at 30 mA. Image J software was used to measure 50 fibers diameters from each SEM photograph.centella, propolis, and hinokitiol incorporated in electrospun nanofibers were analyzed using attenuated total reflectance (ATR) FT-IR spectroscopy . The infrared spectra were recorded from 4000–600 cm−1, with a resolution of 4 cm−1. The infrared beam enters the ATR crystal at an angle of typically 45° and is totally reflected at the crystal to sample interface with a low signal of noise ratio.The presence of centella, propolis, or hinokitiol. The two-dimensional (2D) patterns of nanofibers were recorded by X-ray diffraction equipment (SPring-8 synchrotron radiation facility in Japan) with a wavelength of 0.07085 nm at a 2θ scanning angle between 5–30°. The distance from the sample to the detector (PILATUS 3 × 2 M) was 326.5 mm and the exposure time was 2.0 s. The X-ray diffraction of composite nanofibers was investigated by the MDIP application.Wide-angle X-ray diffraction (WAXD) was used for crystalline structure analysis of PHBH nanofibers with or without centella, PHBH/propolis, PHBH/hinokitiol nanofibers were investigated by using a tensile testing machine  with 50-N load cell at room temperature. Ten specimens were prepared from each composite nanofiber sheet. The specimens with thickness ranging from 0.05 to 0.14 mm were cut into a rectangular shape of 30 mm × 5 mm. The ends of each sample were secured by two clamps with an initial distance of 10 mm and a crosshead speed of 10 mm/min. Tensile strength, Young’s modulus, and elongation at break were calculated on the basis of the stress-strain curves.The mechanical properties of neat PHBH, PHBH/centella, propolis, and hinokitiol was analyzed by a UV-visible spectrophotometer . For release experiments, PHBH composite nanofibers with centella, propolis, or hinokitiol were cut into 2 × 2 cm and measured specimen weight. For each sample, 2 specimens were placed in a test tube filled with 50 mL of phosphate-buffered saline (PBS) (pH 7.4) incubated at 37 °C and stirred at 150 rpm. During the release, 4 mL of the supernatant was retrieved from the release medium at the time points of 5, 10, 20, 30, 40, 50, 60, 120, 240, 360, 480, 600, 720, 1440, 2880, 4320, and 5760 min and the same amount of fresh PBS was added immediately to maintain the medium volume.The release of centella, extract propolis, hinokitiol (0.8 mg) was dissolved in 100 mL ethanol/PBS or methanol/PBS  and then diluted to 400, 200, 100, 50, 25, 12.5, 6.25, 3.125, and 1.56 μg/mL. UV-vis spectrophotometer at a wavelength of 270 nm, 273 nm, 224 nm was used to measure the amount of released centella, propolis, and hinokitiol from PHBH composite nanofibers, respectively. The total amounts of centella, propolis, and hinokitiol from the nanofiber mats were determined as the average value of the three tests. These results were presented in the form of a cumulative amount of release [tM is the amount of centella, propolis, or hinokitiol released at the time t, M∞ is the total amount of centella, propolis, and hinokitiol loaded in PHBH composite nanofibers.The solutions in various concentrations were used to make a standard calibration curve at wavelength 200–700 nm. In detail, certain weight of extract  release :CumulatS. aureus and E. coli were chosen as representatives for gram-positive and gram-negative bacteria. The test method was the disk diffusion test, which has been well established by previous reports [centella, PHBH/propolis, and PHBH/hinokitiol composite nanofibers were placed on the surface of the Petri dishes then they were incubated at 30 °C (E. coli) and 37 °C (S. aureus) for 24 h. The diameters of inhibition zone (mm) were measured for 3 specimens each sample and the results were expressed as mean diameters with standard deviations (in millimeters).To examine the antibacterial activity of natural antibacterial reagents,  reports ,31,32,33p < 0.05 to identify which groups were significantly different from other groups.All experiments were conducted in triplicate and the data are presented as mean ± standard deviation (SD). The significant differences were statistically analyzed by one-way analysis of variance (ANOVA) using R free software. Statistical significance was set at centella solutions , propolis solutions , and hinokitiol solutions  are exhibited in The surface morphology of PHBH nanofibers containing different natural antibacterial products was investigated using SEM. Representative SEM images and the fiber diameter distributions of neat PHBH and PHBH nanofibers loaded with different amounts of centella (15EC and 30EC) and PHBH/methanol–centella (15MC and 30MC) composite nanofibers showed the uniform fiber diameter. These also exhibited the same trend of a decrease in fiber diameter by increasing the ratio of centella solutions (EC and MC) in spinning PHBH solution. The fiber diameter, with high concentrations of centella solution in PHBH/30EC (10%) and PHBH/30MC (10%), diminished to 349 ± 108 nm and 332 ± 62 nm, respectively. This phenomenon may be due to the decrease in the viscosity of polymer solutions by the addition of ethanol or methanol into HFIP. The viscosity of the mixture solution reduced when the considerable amounts of centella solution added into the polymer matrix [centella in PHBH/30EC (10%) (r matrix . InteresEC (10%)  resultedv/v) of 10AP, respectively. This is probably because the evaporation of acetone was faster than ethanol during the electrospinning process.Such a bonded structure was also appeared in PHBH/30EP (7%). The bonded structure in high concentration solutions of EC and EP might be due to using ethanol as the solvent and mixing with HFIP. By contrast, PHBH/30EP (7%) was accompanied by an increase in fiber diameter by loading a high concentration of 30EP. The fiber diameter was increased from 539 ± 99 nm PHBH/30EP (1%) to 739 ± 197 nm PHBH/30EP (7%). A similar behavior was obtained from cellulose acetate nanofibers with a high concentration of honey bee propolis prepared by ethanol solution . Kim et For PHBH/ethanol–hinokitiol (EH) composite nanofibers, the surface morphology of PHBH/30EH (1%) was smooth, homogeneous without beads. The average diameter was 562 ± 87 nm with a majority of fibers in the range of 438 nm to 745 nm. However, it was difficult to obtain uniform nanofibers with high ratios of hinokitiol solutions (EH), more than 1%. As shown in The final surface morphologies of PHBH composite nanofibers were influenced by the kind of used natural products and concentration ratios of natural product solutions to PHBH solutions. This result clearly indicates that the fiber diameters and their distributions can be controlled by concentration ratios of natural products in solutions. In this paper, PHBH composite nanofibers with natural product solutions of PHBH/30EC (10%), PHBH/30MC (10%), PHBH/30EP (7%) and PHBH/30EH (1%) were chosen for further investigation by FTIR, WAXD, mechanical properties, antibacterial test, and release characteristic because these samples had fiber diameter between 300–1000 nm, surface morphology without beads, and high concentration of natural products.−1 were attributed to the stretching vibrations of C–CH3, CH2, CH, and C=O, respectively [The functional groups of natural antibacterial products contained in PHBH composite nanofibers were analyzed by using ATR (FT-IR). The FT-IR spectra of PHBH/30EC (10%), PHBH/30MC (10%), PHBH/30EP (7%), and PHBH/30EH (1%) are shown in ectively .centella powder from ethanol and methanol solutions after filtering and drying exhibit some bands centered at 3320 or 3300 cm−1 corresponding to O–H stretching vibration of carboxylic acid, whereas, 1690 or 1638 cm−1 peaks characterizing C–O stretching vibration. The bands at 1459, 1380, and 1025 cm−1 shows the presence of alkenes with C–H in-plane bending, the stretching vibration of C–N for aromatic amide group, and C–O stretching, respectively . In the case of PHBH/30MC (10%) composite nanofibers, the decline of the band at 3340 cm−1 suggested the formation of the intermolecular hydrogen bond between centella and PHBH polymer.FT-IR graphs of ectively A. These m et al. , and Sugm et al. . As show−1 was assigned to stretching vibration of the O–H group in the phenolic compound and the band at 2920–2870 cm−1 was attributed to C–H aliphatic stretching vibration (stretching vibration of CH2 and CH3) [−1, 1650–1602 cm−1, and 1190 cm−1 were attributed to the C=O group, aromatic ring deformations C=C stretching vibration, and C–O stretching vibration, respectively. In PHBH/30EP (7%) composite nanofibers, some peaks appeared at between 1651 and 1610 cm−1, and 1510 cm−1, confirming the presence of propolis in PHBH nanofiber mats. These bands were assigned to stretching of C=C aromatic ring bands in flavonoids [The FT-IR spectra of propolis powder and PHBH/30EP (7%) nanofibers were exhibited in and CH3) . The banavonoids ,36.−1. These bands represented O–H stretching, C=C and C=O stretching, C=C stretching (in-phase), ring CH bending, C–O–H group, and ring CH bending, respectively [−1) and the C–O–H group (1189 cm−1). The presence of hinokitiol in PHBH composite nanofibers was confirmed by the weak peak appeared at 1610 cm−1 that is related to tropolone carbon ring. In general, the peak intensity decreased by loading natural product might reveal that natural product is homogeneously distributed through the polymer matrix [centella, propolis, and hinokitiol, respectively.The characteristic absorption peaks of hinokitiol powder were observed at 3200, 1609, 1543, 1476, 1459, 1417, 1269, 1185, and 950 cmectively . Dyrskovectively  reportedcentella, propolis, and hinokitiol on the crystallinity of PHBH. All reflections of WAXD and intensity profile are displayed in centella and propolis in PHBH/30EC (10%) and PHBH/30EP (7%) composite nanofibers. Moreover, the calculated crystallinity of neat PHBH (47.0%) was slightly lower than that of PHBH/30EC (55.0%), PHBH/30MC (49.2%), and PHBH/30EP (54.5%) in centella or propolis in nanofibers mat exists.The WAXD measurement was performed in order to confirm the influence of loading centella (EC 10%) and propolis (EP 7%) in PHBH demonstrated the same trend with the tensile strength rose up to 17.8 and 16.4 MPa but it reduced elongation down to 15.6% and 9.3%, respectively. PHBH/30MC (10%) composite nanofibers exhibited no significant differences in tensile strength and Young’s modulus compared to neat PHBH. PHBH/30EH (1%) resulted in a decline in both tensile strength and elongation compared to neat PHBH.Tensile test was conducted on neat PHBH and PHBH composite nanofibers with the natural antibacterial products in order to evaluate the influences of natural product loading. The tensile strength, elongation at break, and Young’s modulus of neat PHBH, PHBH/30EC (10%), PHBH/30MC (10%), PHBH/30EP (7%), and PHBH/30EH (1%) are shown in centella (EC) and propolis (EP) using ethanol as solvent displayed higher tensile strength in comparison to neat PHBH, PHBH/MC, and PHBH/EH. These phenomena may be due to the surface morphology and polymer structure in PHBH composite nanofibers. The surface morphology of PHBH/30EC (10%) and PHBH/30EP (7%) showed bonded-structure between fibers, whereas PHBH/30MC (10%) and PHBH/30EH (1%) surface revealed randomly oriented fibers. Kim et al. [PHBH composite nanofibers with m et al.  reportedHBH/30EC % and PHBm et al. .Additionally, the enhancement of physical properties in PHBH/30EC (10%) and PHBH/30EP (7%) might be related to the rise of the crystallinity of PHBH polymer in PHBH composite nanofibers. It is probably due to the interaction of natural products (EC and EP) to PHBH polymer that was explained by FT-IR and X-ray analysis.centella, propolis, and hinokitol against the gram-positive bacteria S. aureus and gram-negative bacteria E. coli was evaluated using inhibition zone method [S. aureus) and 30 °C (E. coli) for 24 h. centella in PHBH/30MC (10%) demonstrated low antibacterial activity against S. aureus with inhibitory zone of 7.7 mm and no effect against E. coli. The diameter of the inhibition zone of PHBH/30EP (7%) was 18.3 mm for S. aureus and 17.3 mm for E. coli. PHBH/30EH (1%) composite nanofibers have extensive inhibition zone against S. aureus (25.7 mm) and E. coli (29.7 mm), respectively. The zones of inhibition for PHBH/30EP (7%) against S. aureus were slightly broader than that against E. coli. Zeighampour et al. [The antibacterial activity of neat PHBH nanofibers, PHBH composite nanofibers with e method ,33,40,41r et al.  explainecentella. However, the low effect antibacterial activity of centella (EC or MC) might be due to the low chemical content in PHBH composite nanofiber as analysed by FT-IR, which PHBH/30EC (10%) showed no clear detection of asiatic acid. For further work, raising the centella concentrations in ethanol or methanol solutions might be one of the solutions to increase the antibacterial activity, if those will be used for wound healing application. Overall, the natural products  loaded into PHBH composite nanofibers inhibited the growth of S. aureus and E. coli powerfully.Regarding these results, the antimicrobial effects of PHBH composite nanofibers with propolis and hinokitiol were observed to be better than these of centella, propolis, and hinokitiol from electrospun PHBH composite nanofibers were plotted as a function of time in PBS with pH 7.4 at 37 °C. The release curves of PHBH/30EC (10%), PHBH/30MC (10%), PHBH/30EP (7%), and PHBH/EH (1%) composite nanofibers demonstrated different release behaviors in each sample as shown in Centella from PHBH/30EC started to release within the 10 min and completely released in 20 min. In the case of PHBH/30MC, centella from PHBH/30EC and PHBH/30MC were 13.9 % and 31.5%, respectively. While the significant release amounts of propolis from PHBH/30EP started to be detected after 10 min and gradually increased up to 2880 min (48 h) with the release amount of 9.6%. The release of propolis from PHBH/30EP sustained longer than others but the release amount of propolis was less. The fast release of hinokitiol from PHBH/30EH was noticeable in the first 5 min and gradually increased up to 20 min with a maximum release of 46.1%. Afterwards, the release of hinokitiol gradually declined over 240 min (4 h). This decline might be due to the degradation of hinokitiol in PBS solution by light, heat, or solvent (ethanol). In the study of ciprofloxacin hydrochloride (CpHCl) release from electrospun alginate [The release profiles of alginate , at aroualginate  reportedEspecially for PHBH/30EC (10%) and PHBH/30EP (7%), those have similar release behaviors, when only small amounts of natural products were released to PBS. It can be considered due to the high crystallinity of those PHBH composite nanofibers that affected release characteristics. In comparison, PHBH/30EH (1%) with low crystallinity showed fast and high release amounts. Study of drug release of ampicillin incorporated poly(methyl methacrylate)–nylon6 core/shell nanofibers proved that the increase of released drug, most probably due to decreased crystallinity in the polymer matrix .centella, propolis, and hinokitiol) loaded into PHBH composite nanofibers were successfully fabricated by the electrospinning process. The fiber diameter and surface morphology of PHBH composite nanofibers with antibacterial reagents can be controlled by different kinds of natural antibacterial products, solvent systems for dissolution, and component concentrations in the PHBH-HFIP polymer solutions. The presence of carboxylic acid and aromatic amide groups in PHBH/centella, aromatic ring bands due to flavonoid in PHBH/propolis, and tropolone carbon ring in PHBH/hinokitiol were confirmed by FT-IR. The loading of centella and propolis led to an increase in the crystallinity of the PHBH polymer. Furthermore, the loading of centella and propolis improved the tensile strength, compared to neat PHBH nanofibers. Hinokitiol and propolis were proved to be potent antibiotics by large inhibition zones against both E. coli and S. aureus. The release of centella and hinokitiol from PHBH nanofibers was fast and finished in 20 min with maximum release of 13.9% (PHBH/30EC), 31% (PHBH/30MC), and 46% (PHBH/30EH). Whereas, the release of propolis was continuously to 48 h and maximal release was 9.5%. These results in our study indicated that natural antibacterial products loaded into PHBH composite nanofibers can improve the mechanical properties (PHBH/30EP and PHBH/30EC) and prove the composite nanofibers antibacterial effects against the gram-negative and gram-positive bacteria (PHBH/30EP and PHBH/30EH), which are important characteristics as wound healing materials. Interestingly, PHBH/30EC (10%) showcased good mechanical properties and PHBH/30MC (10%) exhibited good morphology in nanofiber form, however, it may necessary to increase the concentration of centella in centella solution (ethanol or methanol) if it is to be used as antibacterial reagents. PHBH/30EH (1%) might be used as drug delivery with rapid release but possesses low mechanical properties. PHBH/30EP (7%) could be used in wound healing with needed mechanical properties and slow release for long-time effects.The natural antibacterial products (


Control over spin states at the single molecule level is a key issue in the emerging field of molecular spintronics. Here, we explore the chemical adsorption effect on the magnetic and spin-transport properties of individual magnetic molecule by performing extensive density functional theory calculations in combining with non-equilibrium Green’s function method. Theoretical results clearly reveal that the molecular magnetic moment of Mn-salophen can be effectively tuned by adsorbing F and CO on the central Mn cation, while the adsorbed NO molecule quenches the molecular magnetic moment. Without chemical adsorption, the currents through Mn-salophen molecular junction just show a little distinction for two spin channels, which agrees well with previous investigation. Remarkably, the conductive channel can be switched from the spin-up electrons to the spin-down electrons via adsorbing F and CO, respectively, and the corresponding two Mn-salophen molecular junctions with chemical modifications display nearly perfect spin-filtering effect. The observed spin switch and the predicted spin-filtering effect via chemical adsorption indicates that Mn-salophen holds potential applications in molecular spintronic devices. Molecular spintronic devices based on the magnetic molecule as the functional units, such as molecular rectifier, molecular switch, and molecular transistor, have already attracted a great deal of attention in recent years since it holds promise for the next generation of electronic devices with enhanced functionality and improved performance ,4,5,6,7., and O2 ,14,15,16, and O2 . Kondo end gases . Clearly3d transition metal(II) salophens , and found that only Co-salophen junction display robust spin-filtering effect [Molecular spin filter has been demonstrated to be an important spintronic device with conducting for one spin direction and insulating for the opposite direction. Till now, nearly perfect spin-filtering effect has been observed in many molecular junctions based on various molecules, including MnCu single-molecule magnet ,20, spinomplexes ,22,23, momplexes ,25, Fe-complexes , and Eu-omplexes . We noteoperties ,29, but g effect .Here, we revisit Mn-salophen and try to explore the chemical adsorption effect on the magnetic and spin-transport properties of Mn-salophen sandwiched between two Au(100) electrodes by performing extensive density functional theory calculations combining with non-equilibrium Green’s function method. According to the calculated molecular magnetic moments, we find that the spin states of Mn-salophen can be easily controlled via chemical adsorption. Remarkably, upon the F and CO adsorption on the central Mn cation, two molecular junctions can act as efficient spin filters, and the corresponding conductance is dominated by the spin-up and spin-down electrons, respectively. The observed nearly perfect spin-filtering effect and the switching between two spin channels implies that Mn-salophen is a promising candidate for designing molecular spintronic devices.Here, geometry optimizations, electronic structures and transport properties of Mn-salophen with F, CO, and NO chemical adsorptions are calculated by using ATK package ,32,33, wBefore examining the chemical adsorption effect, we perform benchmark spin-polarized DFT calculations for free Mn-salophen molecule. The molecular structure of Mn-salophen is illustrated in the top panel of Previous investigations have demonstrated that the spin-states of individual magnetic molecule  can be reversibly controlled by single H atom adsorption  and spin3d electrons in Mn-salophen, and the corresponding molecular MM is 3.0 3d electrons of Mn ion transfers to F atom, the atomic MMs of Mn and F atoms are predicted to be 1.90 and 0.07 3d electrons. The atomic MM of Mn, C and O atoms in CO is about 1.01, −0.08 and −0.02 Please note that the MM of Mn-salophen molecular junction (3.0 The obtained current-voltage (I-V) curves for four Mn-salophen molecular junctions under the bias voltage range from 0.0 to 0.8 V are plotted in (1) Without chemical adsorption, the currents through Mn-salophen molecular junction for the spin-up and spin-down electrons just show a little distinction. For example, at 0.4 V, the calculated Is report .(2) Upon F adsorption on the central Mn atom, the current of the spin-down electrons (I(3) As for CO adsorption, we find that the conductance through molecular junction is dominated by the spin-down electrons. The calculated (Itetragon .(4) When NO connecting to Mn ion, the ITo explore the nature of the above dramatically different characteristics of the I-V curves of four different molecular junctions, we calculate the zero-bias spin-polarized transmission spectra, and plot them in Upon F adsorption, the overall features of transmission spectra of Mn-salophen molecular junction significantly changed, as shown in omplexes ,37.28 , and Fe2omplexes ,23.As shown in Next, we turn to analyze the spin-polarized electronic structures of Mn-salophen molecule with F, CO, and NO adsorption, as shown in In general, standard GGA functionals  cannot predict correct HOMO-LUMO gaps for metal-containing molecules and some corrections, such as GGA+U, is necessary. ,40 Here,In summary, combining extensive density functional theory calculations with non-equilibrium Green’s function method, we explore the spin states and spin-transport properties of Mn-salophen sandwiched between Au electrodes. We find that the molecular MM of Mn-salophen can be effectively tuned via chemical adsorption. Without chemical adsorption, the currents of the spin-up electrons through Mn-salophen molecular junction is close to that of the spin-down electrons. Remarkably, theoretical results clearly reveal that the conductive channel can be switched from the spin-up electrons to the spin-down electrons via adsorbing F and CO, respectively, and the corresponding two Mn-salophen molecular junctions display nearly perfect spin-filtering effect. The observed spin-filtering effect and the switching between the spin-up and spin-down channels via chemical adsorption indicates that Mn-salophen holds potential applications in molecular spintronic devices.


Acremonium. Here, we present the asymmetric total synthesis of acremine F which hinges on a modestly enantioselective dihydroxylation and a subsequent kinetic resolution via a highly selective asymmetric reduction. Chemoselective oxidation of acremine F gave access to acremines A and B. The dimerization of acremine F to bisacremine E was investigated but could not be achieved, shedding light on the formation of the acremine dimers in nature.The acremines are a family of meroterpenoids isolated from fungi of the genus  Acreonium byssoides, isolated from grapevine leaves that were artificially inoculated with Plasmopora viticola (5) exhibited no significant bioactivity, acremines A–D showed inhibition of P. viticola sporangia germination.Endophytic fungi grow in a symbiotic relationship with their plant hosts , which iviticola  3]. Thi. ThiAcreA. persicinum SC0105 [5) units by a formal [4 + 2] cycloaddition followed by condensation and oxidation.In 2015, Wei and co-workers discovered bisacremines E–G, the most complex members of the acremine family, from the soil-derived strain m SC0105 . These nGiven the diversity and structural beauty of this class of natural products, it is not surprising that the acremine family has attracted the attention of the synthetic community –8. Never5 is depicted in 9. Compound 9 in turn could be traced back to silyl enol ether 10. Ent-10 was first reported by Herzon and co-workers  cycloaddition involving two acremine F (5) units [5 through a Diels–Alder cycloaddition is not electronically favorable, we speculated that this reaction might proceed through ionic intermediates  relying on the combination of a modestly enantioselective oxidation and a highly enantioselective reduction with kinetic resolution to access the acremine framework. The route proved to be scalable and delivered 300 mg of the natural product. Acremine F could further be converted into acremines A (1) and B (2) by a selective oxidation providing a versatile entry into this class of natural products. Furthermore, we investigated the proposed biomimetic dimerization of 5 to bisacremine E. Since these extensive studies were unsuccessful and no dimers could be observed under a variety of biomimetic conditions, it appears that this transformation requires enzymatic catalysis in nature.In conclusion, we report the first asymmetric synthesis of acremine F (16 and 17. CIF files for 16 (CCDC 1854563) and 17 (CCDC 1854564) are available free from charge on https://www.ccdc.cam.ac.uk/structures/).Experimental procedures, spectroscopic data and copies of NMR spectra (PDF) as well as crystallographic data of compounds File 1Experimental part.


Levels exceeding the standard reference interval (RI) for total thyroxine (TT4) concentrations are diagnostic for hyperthyroidism, however some hyperthyroid cats have TT4 values within the RI. Determining outlier TT4 concentrations should aid practitioners in identification of hyperthyroidism. The objective of this study was to determine the expected distribution of TT4 concentration using a large population of cats  of unknown health status to identify unexpected TT4 concentrations (outlier), and determine whether this concentration changes with age.This study is a population-based, retrospective study evaluating an electronic database of laboratory results to identify unique TT4 measurement between January 2014 and July 2015. An expected distribution of TT4 concentrations was determined using a large population of cats  of unknown health status, and this in turn was used to identify unexpected TT4 concentrations (outlier) and determine whether this concentration changes with age. All cats between the age of 1 and 9 years  had the same expected distribution of TT4 concentration (0.5–3.5ug/dL), and cats with a TT4 value >3.5ug/dL were determined to be unexpected outliers. There was a steep and progressive rise in both the total number and percentage of statistical outliers in the feline population as a function of age. The greatest acceleration in the percentage of outliers occurred between the age of 7 and 14 years, which was up to 4.6 times the rate seen between the age of 3 and 7 years.TT4 concentrations >3.5ug/dL represent outliers from the expected distribution of TT4 concentration. Furthermore, age has a strong influence on the proportion of cats. These findings suggest that patients with TT4 concentrations >3.5ug/dL should be more closely evaluated for hyperthyroidism, particularly between the ages of 7 and 14 years. This finding may aid clinicians in earlier identification of hyperthyroidism in at-risk patients. The diath percentile) and upper quartile (75th percentile). The expected distribution of TT4 concentration is defined as those contained within an upper value (1.5*IQR+ upper quartile) and a lower value (1.5*IQR-lower quartile). An unexpected TT4 concentration signifies an outlier, even if the value is within the laboratory’s RI, and is defined as a data point that is outside of the expected concentration for the sampled population [In studies where the health of the patients cannot be established, the data is often evaluated using Tukey’s method , which ipulation –10.Clinicians frequently encounter patients with compatible physical examination findings (enlarged thyroid) and/or clinical signs of hyperthyroidism but with TT4 concentrations within the laboratory’s RI. Such cats may be truly normal (non-hyperthyroid) or abnormal  requiring additional conformational testing or monitoring . The ideThe objectives of the study reported here were to 1) Determine the expected distribution of TT4 concentration and outlier values based on a large population of cats encompassing all ages and breeds in the US, 2) compare these TT4 concentrations to the standard laboratory RI, and 3) determine if and when the unexpected TT4 concentration changes with age.Retrospective, population-based review and analysis of laboratory results in adult cats for which a unique TT4 concentration was measured.An electronic database of laboratory results from a large reference diagnostic laboratory  was reviewed between January 2014 and July 2015 from both general practice and specialty veterinary hospitals in the US. Data provided was anonymized with each patient being assigned a unique identification number based on the patient’s name and veterinary clinic. Information obtained included TT4 result and patient signalment . Client information was not included. If multiple TT4 concentrations were measured in a given patient in the same hospital during the study period, only the initial measurement was included to assist in excluding subsequent samples from ill patients with multiple laboratory submissions and hyperthyroid cats with multiple TT4 measurements following the institution of therapy. Patients between the age of 1 week and 1 year were categorized as < 1 year of age. TT4 concentrations from cats over the age of 20 years, TT4 concentrations greater than 20 ug/dL, and samples for which there was no assigned age or sex were excluded from the data set due to their low frequency and/or high likelihood of error. All age groups were included to assess the effect of age on both expected and unexpected TT4 concentrations and to document if and when the unexpected TT4 concentration change with age.15 and Long Island, NY)[th percentile interval  for results from > 1000 clinically normal cats ranging in age from 1–3 years collected as part of a wellness study performed by Antech Diagnostics.All serum TT4 concentrations were measured by the same homogenous thyroxine enzyme immunoassay , which has been previously validated for measuring TT4 concentrations in cats –16. GuidIn the Long Island, NY laboratory  the follth percentile) and upper quartile (75th percentile). The expected distribution of TT4 concentration was defined as being within an upper limit of 1.5*IQR + upper quartile and a lower limit of 1.5*IQR–lower quartile. The Tukey method does not make assumptions about how the population of values is distributed or its mean or standard deviation. “Fences” using multipliers of 1.5 or 3.0 of the interquartile range are termed outliers or extreme outliers respectively. Since it is better for sensitivity of a test to error on being wrong (less specific) than the other way around we used the same thought in choosing the multiple 1.5*IQR- 25% quantile and 1.5*IQR +75% quantile rather than 3.0*IQR.Statistical analysis was performed using JMP 11.1 software program from SAS . A search was performed to identify data points according to the selection criteria described above. As the data was not normally distributed as determined by the Kolmogorov-Smirnov test, and the health of the patients was not established, the data was evaluated using Tukey’s method , which iValues below 0.5 ug/dL were automatically assigned a value of 0.5 ug/dL owing to the lower limit of detection of the TT4 assay. We had 14,463 values reported as < 0.5 ug/dl. This represented ~ 2.5% of the dataset. SAS has found that when dealing with values comprising less than 5% of a dataset, the method of handling limit of detection values does not affect the analysis. When, we evaluated these values by reporting them as either 0.5 ug/dl or as 0.25 ug/dl there was no difference in the results. Outliers were represented by all values outside of the expected TT4 concentration.The number of outliers in each age group was determined by totaling the quantity of TT4 concentrations above 3.5 ug/dL. The percentage of outliers in each age group was determined by calculating the number of TT4 concentrations greater than 3.5 ug/dL divided by the total number of TT4 concentrations in the respective age group. The rate of change (slope) of percentage of unexpected outliers was determined by calculating the change in percentage outliers divided by the change in age group categories.Using the search criteria above, an initial 543,861 TT4 concentrations were identified between January 2014 and July 2015. Data were compiled from a total of 8,673 hospitals in the US. Samples were categorized into the following groups based on sex and reproductive status; female intact , female spayed , male intact , male neutered . Of the total samples tested, 485,195 samples comprising 89% of the total reference values were categorized as domestic short hair, domestic medium hair, domestic long hair, not identified, unknown, or blank. The remaining 58,666 samples tested comprising 11.0% of the total reference values were represented by 41 different Cat Fanciers’ Association breeds. TT4 concentrations greater than 20 ug/dL , TT4 concentrations belonging to cats older than 20 years of age , and TT4 concentrations for which there were no assigned age or obvious misinformation  were excluded from the data set leaving a total of 531,765 unique TT4 concentrations. In addition, removing cats > 20 years and those with results > 20 ug/dl did not change any of the data conclusions regarding upper fences or outliers.The data was broken down into age categories including <1 year and years 1 through 20. For each age group, the median value, lower and upper quartiles, IQR, expected TT4 concentration, and outlier values were determined .For all cats in the data set, the median TT4 concentration was 1.95 ug/dL and the expected distribution of TT4 concentration was 0.5–4.3 ug/dL. For all cats between the age of 1 and 9 years  the median TT4 concentration, lower and upper quartiles, IQR, expected TT4 concentration, and outlier values were identical .rd degree polynomial distribution y = -0.0001x3 + 0.0046x2–0.0292x + 0.0666; R2 = 0.9956. Thus, 99.56% of the variation of values outside of 3.5 ug/dL can be explained as a function of age.These values were determined to be; median 1.9 ug/dL, lower quartile 1.5 ug/dL, upper quartile 2.3 ug/dL, IQR 0.8, expected TT4 concentration 0.5–3.5 ug/dL, and outlier > 3.5 ug/dL. In cats <1 year of age, the values were determined to be; median 1.8 ug/dL, lower quartile 1.4 ug/dL, upper quartile 2.4 ug/dL, IQR 1.0, expected TT4 concentration 0.5–3.9 ug/dL, and outlier > 3.9 ug/dL. As age increased above 9 years, the distribution of values shifted to the right as these populations contained a greater percentage of cats with higher TT4 concentrations. The number of cats in each age group that had a TT4 concentration greater than 3.5 ug/dL, defining a statistical outlier, as well as the percentage of outliers calculated as the number of outliers divided by the total number of cats of a particular age group were determined . The numThere were 3 age groups for which the rate of change (slope) of unexpected outliers were identifiable, including 3–7 years, 7–14 years, and 14–20 years. The rate increase was highest from 7–14 years where the slope was 4.6 times faster than from 3–7 years of age. Cats 14–20 years of age had a rate increase that was 1.5 times faster than young cats but 3 times slower than cats aged 7–14 years .This study differs from previous studies by examining a large population of cats of all ages throughout the United States. As the data was not normally distributed and the health of the patients was not established, the data was evaluated using Tukey’s method which is appropriate for analyzing large univariate data sets and such approaches are distribution independent. By defining exclusion bounds (outliers) rather than the inclusion bounds determined by RI’s, we are able to identify those patients at risk of hyperthyroidism even when their TT4 values fall within the laboratory’s RI.15 and a similar RI of 0.8–3.8 ug/dl found when evaluating 131 healthy, euthyroid cats, over the age of 7 years [By examining a total of 531,765 unique TT4 concentrations that were measured using the same validated immunoassay it was shown that for all cats in the data set, the median TT4 concentration was 1.95 ug/dL and the expected distribution of TT4 concentration was 0.5–4.3 ug/dL. This is very similar to the laboratory’s RI of 0.8–4.0 ug/dl which was determined in >1,000 healthy cats between the ages of 1–3 years 7 years . This hiPerhaps the most notable finding in this paper was that cats between 1–9 years of age had an identical expected distribution of TT4 concentration (0.5–3.5 ug/dL). Cats with TT4 concentrations greater than 3.5 ug/dL represented unexpected outliers in the population even though TT4 concentrations between 3.5–4.0 ug/dl are considered to be within the established RI for the laboratory. Based on this information, the finding of a TT4 between 3.5–4.0 ug/dl, along with historical and physical examination findings suggestive of hyperthyroidism, indicates the need for additional testing or monitoring.A previous study by Peterson et al  which co19 In veterinary medicine, RIs are generally based on a relatively small sample size. Although there are conventional methodologies for establishing RIs using a small number (i.e. <120) of reference values [The development of RIs in a given population was a concept first introduced in human medicine and later applied to veterinary medicine –21. Laboe values ,22, the e values .th through 97.5th percentile range) and the expected TT4 concentrations in the 531,765 cats was determined using Tukey’s method which can be used for data sets where the health status of the population is unknown such as determination of RIs in wildlife species [The expected TT4 values were determined using non-parametric methods of percentile estimates with confidence intervals to determine the central 95th percentile interval  [An assumption was made about the accuracy of the reported age, sex, reproductive status, and breed by the submitting hospital. While there are recording errors that do occur, the signalment corresponding to the individual data points was accepted to be accurate in the current study. Moreover, while there may have been errors in TT4 concentrations due to pre-analytical errors , analytical errors, or post-analytical (transcription) errors , the TT4c –16,26 att assay) –29. Despt assay) –31, in oDiagnosis of feline hyperthyroidism relies on compatible clinical signs , identification of a thyroid nodule, and an elevation in circulating thyroid hormone levels. Treatment of hyperthyroidism regardless of modality  carries a favorable prognosis ,32–36. TThis study demonstrated that age was a strong predictor for the number and percentage of unexpected outliers in the population, accounting for 99.5% of the variation in outlier TT4 concentrations. The greatest change in the number and percentage of outliers occurred in cats aged 7–14 years, which suggests that cats in this age group are more likely to become outliers, and thus should be more closely scrutinized for abnormal TT4 concentrations and clinical signs of hyperthyroidism compared to cats of other age groups. These results are consistent with the observation that hyperthyroidism occurs most commonly in older cats ,24.Since its first description in 1979, the incidence of hyperthyroidism has dramatically increased prompting veterinarians and researchers to hypothesize whether exposure to environmental thyroid-disruptor chemicals or other environmental, genetic or dietary factors are involved in the pathogenesis of hyperthyroidism , fertiliBased on the current study, the identification of a TT4 concentration greater than 3.5 ug/dL is an unexpected finding and characterizes an outlier in the general population. Irrespective of clinical signs of hyperthyroidism, the identification of an outlier warrants suspicion for the possibility of hyperthyroidism. If the TT4 concentration is greater than 3.5 ug/dL but less than the upper limit of the reporting laboratory’s RI, and the patient exhibits clinical signs of hyperthyroidism, further evaluation is warranted to rule in or rule out hyperthyroidism such as by measurement of free T4 by equilibrium dialysis (fT4ED) or thyroid stimulating hormone (TSH) where a normal TSH concentration can be used to rule-out hyperthyroidism . The ideWith the establishment of this new expected TT4 concentration in the cat, future studies will focus on determining how breed and geographical distribution within the US affects the expected TT4 concentration. It would be of interest to determine the true incidence and prevalence of hyperthyroidism in the US and different parts of the world, and to evaluate changes in prevalence over time by repeating this study several years from now to determine if the expected TT4 concentration or outlier population changes.


N-acetylglucosaminidase, has a unique character of a wide optimal pH range of pH 5.0–7.0. We hypothesized that this endoglycosidase degrades keratan sulfate (KS) polymer in circulating blood and, therefore, ameliorates the accumulation of KS in multiple tissues. We propose a novel approach, Substrate Degradation Enzyme Therapy (SDET), to treat bone lesion of MPS IVA. We assessed the effect of thermostable keratanase on blood KS level and bone pathology using Galns knock-out MPS IVA mice. After a single administration of 2 U/kg (= 0.2 mg/kg) of the enzyme at 8 weeks of age via intravenous injection, the level of serum KS was significantly decreased to normal range level, and this suppression was maintained for at least 4 weeks. We administered 2 U/kg of the enzyme to MPS IVA mice every fourth week for 12 weeks  at newborns or 8 weeks of age. After a third injection, serum mono-sulfated KS levels were kept low for 4 weeks, similar to that in control mice, and at 12 weeks, bone pathology was markedly improved when SDET started at newborns, compared with untreated MPS IVA mice. Overall, thermostable keratanase reduces the level of KS in blood and provides a positive impact on cartilage lesions, demonstrating that SDET is a novel therapeutic approach to MPS IVA. Mucopolysaccharidosis IVA (MPS IVA) is caused by a deficiency of the lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase (GALNS). Conventional enzyme replacement therapy (ERT) is approved for MPS IVA. However, the fact that the infused enzyme cannot penetrate avascular lesions in cartilage leads to minimal impact on the bone lesion. Moreover, short half-life, high cost, instability, and narrow optimal pH range remain unmet challenges in ERT. Thermostable keratanase, endo-β- Mucopolysaccharidosis type IVA  is an autosomal recessive disorder caused by a deficiency of the lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase (GALNS) ,5,6. GALEnzyme replacement therapy (ERT) and hematopoietic stem cell transplantation (HSCT) coupled with various surgical interventions are treatment options currently available for patients with MPS IVA in clinical practice. ERT and HSCT are based on the principle of cross-correction that lysosomal enzymes are uptaken into cells via the mannose-6-phosphate receptor. ERT for MPS IVA was approved in 2014 by the Food and Drug Administration; however, several limitations are indicated: (1) weekly infusions for 5–6 h, (2) high cost ,13, and HSCT for MPS IVA was approved in Japan in 1995 with the initial success of transplantation, and at least four patients have been followed over 15–25 years . Only onN-acetylglucosamine 6-sulfatase, β-N-acetylhexosaminidase, and β-galactosidase. These exohydrolases degrade GAG in lysosomes within the cell since the lysosomal enzymes are active only in an acidic environment with narrow pH range from 4.5 to 5.0. Current ERT does not directly degrade accumulated KS in blood and ECM in multiple tissues, including bone and cartilage lesions.In addition, recent studies have indicated that not only accumulated GAG within cells but that in the neighboring ECM provide a critical role to lead to abnormal chondrogenesis and endochondral ossification ,26,27,28N-acetylglucosaminidase from Bacillus circulans and have identified a characterization of thermostability in this enzyme. Thus we named this enzyme, thermostable keratanase . Chondrosine was used as an IS.Serum and tissue GAG level were measured by LC-MS/MS as described previously ,48,49,50Toluidine blue staining and hematoxylin and eosin staining were performed as previously described . Brieflyp < 0.05.All data were expressed as means and standard deviations (SD). Two comparison tests and multiple comparison tests were performed by unpaired t-test or one-way ANOVA with the Bonferroni’s post-hoc test using GraphPad Prism 5.0 , respectively. The statistical significance of difference was considered as


The limitations of conventional therapeutic drugs necessitate the importance of developing novel therapeutics to treat diverse diseases. Conventional drugs have poor blood circulation time and are not stable or compatible with the biological system. Nanomaterials, with their exceptional structural properties, have gained significance as promising materials for the development of novel therapeutics. Nanofibers with unique physiochemical and biological properties have gained significant attention in the field of health care and biomedical research. The choice of a wide variety of materials for nanofiber fabrication, along with the release of therapeutic payload in sustained and controlled release patterns, make nanofibers an ideal material for drug delivery research. Electrospinning is the conventional method for fabricating nanofibers with different morphologies and is often used for the mass production of nanofibers. This review highlights the recent advancements in the use of nanofibers for the delivery of therapeutic drugs, nucleic acids and growth factors. A detailed mechanism for fabricating different types of nanofiber produced from electrospinning, and factors influencing nanofiber generation, are discussed. The insights from this review can provide a thorough understanding of the precise selection of materials used for fabricating nanofibers for specific therapeutic applications and also the importance of nanofibers for drug delivery applications. Nanotechnology—the use of nanomaterials for biomedical applications—is an emerging and promising paradigm in biomedical research. Nanomaterials with exceptional physiochemical properties, biocompatibility and minimal biological toxicity, can sense local biological environments and initiate cellular level reprogramming to achieve the desired therapeutic efficacy. Diverse zero dimensional , one-dimensional  and two-dimensional  nanomaterials along with nanosized particles are currently used for diagnosis, imaging and therapy ,2,3. NanElectrospinning is a unique and versatile technique that depends on the electrostatic repulsion between surface charges to constantly draw nanofibers from viscoelastic fluids. Polymers, ceramics, small molecules and their combinations are used as rich materials for the production of nanofibers. In addition to solid nanofibers, a secondary structure of nanofibers—including porous, hollow or core-sheath structures—has been manufactured and the surface of the structure can be functionalized with different molecular moieties, during or after the electrospinning process . ElectroWe have reviewed the most efficient and recent therapeutic applications of electrospun based nanofibers and their future perspectives. Different electrospinning techniques are described here, along with their biomedical applications and suitability. Nanofibers are generally natural or synthetic polymers and they have vast therapeutic applications even though only a limited number reach clinical trials. The application of nanofibers as drug delivery carriers—especially for cancer drugs, antibacterial drugs, nonsteroidal anti-inflammatory drugs, cardiovascular agents, gastrointestinal drugs, antihistamine drugs, contraceptive drugs and palliative drugs delivery—is described here. Nanofibers have promising therapeutic applications for DNA, RNA and growth factors, for example, protein and steroid hormones delivery, which are included in this review. The current scenario for nanofiber based materials is mostly promising for wound dressing, sensor technology and for the catalysis of various reaction pathways in the laboratory . The theElectrospinning is considered a promising, highly productive and simple method for fabricating the nanofibers of polymers, composites, and inorganic materials, including carbides, oxides, nitrides and hybrid composites. In the electrospinning technique, electrostatic forces are utilized to produce nanofibers from a polymer solution . In geneAlthough fibers produced by different electrospinning methods have attracted increasing attention in the field of biomedical applications, challenges persist for the selection of an appropriate method, as well as for optimizing multiple parameters to generate robust cargo loaded nanofibers . ElectroBlend electrospinning is the conventional electrospinning technique, in which the drug/biomolecules are dispersed or dissolved in the polymer solution, resulting in the development of nanofibers with drugs/biomolecules dispersed throughout the fibers . AlthougCo-axial electrospinning is an improvement on conventional blend electrospinning, in which two nozzles are connected to the high voltage source rather than one nozzle. Two different solutions are loaded within each nozzle and are pumped out to generate nanofibers with core-shell morphologies . Both syEmulsion electrospinning requires a setup similar to that of blend electrospinning, in which two immiscible solvents are simultaneously spun to generate core-shell nanofibers. Here, active bioactive molecules, along with surfactants, are initially allowed to form W/O emulsions and are later mixed with the polymer matrix solution . During Melt electrospinning has gained more attention in the electrospinning field, where toxicity and solvent accumulation is a concern. In this process, polymer melt is used instead of a solution, which is transformed from liquid to solid to achieve the desired product on cooling, rather than through solvent evaporation . The polGas jet electrospinning is an improvement on the conventional melt electrospinning technique, in which the conventional electrospinning setup is additionally equipped with a gas jet device. The major limitation of melt electrospinning is that it requires definite control over the temperatures and therefore multiple heating zones must be placed to maintain the polymer melt. This results in generation of thicker nanofibers compared to the solutions. In this technique, the co-axial jet is surrounded by a tube feeding the heated gas, which can provide sufficient heat near the nozzle and delay the process of polymer solidification. For example, Zhmayev et al. spun polylactic acid (PLA) and showed a decrease in the diameter of the nanofibers obtained by gas jet electrospinning compared to normal electrospinning. Interestingly, the heated gas flow-rate had a significant effect on the diameter of the nanofiber. The increased gas flow rate can offer additional drag force to the jet surface which can result in the development of thinner nanofibers. Zhmayev et al.’s result showed a significant decrease in the diameter of the nanofiber, from 350 nm to 183 nm, when the gas flow rate was increased from 5.0 L/m to 15.0 L/m ,40.Most conventional drugs are hydrophobic and suffer from poor bio-distribution, solubility and stability in the biological system. Moreover, these drugs do not have the desired active targeting capabilities, which can result in non-specific systemic toxicity or faster elimination from the body, without achieving the desired therapeutic efficacy. Drug delivery systems (DDS) are approaches, formulations and technologies for transporting therapeutic agents to the targeted therapeutic site in the body . The devOf all delivery routes, the oral route is considered the most preferable and convenient route of administration, which can overcome the problems associated with other routes of administration . HoweverThe oral mucosa has its own subdivisions according to its different regions, namely, buccal, sublingual and gingival. A few years ago, researchers were also interested in designing nanofiber based drug delivery systems for sublingual (under the tongue) or buccal (between the gums and teeth) routes of administration. The sublingual or buccal delivery systems usually allow the drug-loaded electrospun nanofibers to dissolve in the presence of mucus so that the drug can directly penetrate into the small blood vessels. Interestingly, these oromucosal routes of administration are the most studied sites for nanofiber-based therapeutics, offering versatile and multifunctional drugs, DNA, RNA, protein, peptide, growth factors or vaccine delivery platforms . Pediatric patients (aged under 6 months) usually cannot swallow any drug or food supplement. Under such circumstances, the rectal route could be the best alternative to oral drug delivery systems. In addition, the rectal route is also effective in cases of unconscious or vomiting patients. An electrospun nanofiber-based rectal drug delivery system is continually gaining popularity. In the treatment of post-operative peritoneal effusion following rectal/pelvic surgery, electrospun nanofibers could be considered a safe potential biocompatible, and biodegradable sealing fiber. The first clinical trial on the safety and sealing properties of electrospun nanofibers in lymphorrhea following pelvic surgery was published in 2014 . The autElectrospun nanofiber scaffolds targeting vaginal routes have recently been investigated. However, the acidic condition of vaginal mucosa (~pH 4) should be considered before designing vaginal drug delivery systems using nanofibers. For example, Brako et al. fabricated progesterone-encapsulated nanofibers targeting the vaginal route of administration. The authors used a mucoadhesive molecule (carboxymethylcellulose) for the fabrication of electrospun nanofibers. As-synthesized progesterone/carboxymethylcellulose electrospun nanofibers exhibited sustained release properties ,55. AnotCurrently, supramolecular peptides are used to fabricate electrospun nanofiber scaffolds. Electrospun supramolecular peptide nanofibers have proven their potential for intranasal vaccine delivery. For example, Si and coworkers developed intranasal administrated influenza vaccine using peptide nanofibers consisting of virus polymerase. This self-assembled nanofiber could elicit both humoral and cell mediated immune responses against the influenza virus. To be specific, nasal administrated nanofiber vaccine particles were initially ingested by antigen presenting cells in lung-draining mediastinal lymph nodes and activated both Th1 Th2 to produce the antibody against the given antigen. Furthermore, a high immune response was elicited without any use of vaccine adjuvant .Generally, various drops or ointments are used for ocular infection or inflammation. In these cases, those drugs are easily excreted from the eye very quickly. Therefore, to improve therapeutic efficacy and overcome the repetitive administration of an established ocular dosage, nanofiber-based ocular inserts have been considered and compared with eye drops or similar methods of dosage. Drug-loaded nanofiber scaffolds that allow the controlled release of incorporated drugs can be placed into the ocular mucosa. For example, the use of brimonidine tartrate (BT)-loaded electrospun nanofibers as ocular inserts were used to treat ocular infection and inflammation, which is described in detail in Transdermal drug delivery systems (TDDS) deliver a drug locally, avoiding undesired drug distribution and possessing excellent skin permeability. Due to high solubility, great morphology and sustained drug release kinetics, electrospun nanofibers have been developed into TDDS. It has been reported that electrospun nanofibers could enhance solubility and form a transdermal patch for the biopharmaceutics classification systems II drugs with a high therapeutic efficiency and no cytotoxicity. Hydrophilic polymers with great permeation are the strength of TDDS, and several in vitro and in vivo evaluations suggest the possibility of using drug-loaded electrospun nanofibers in establishing TDDS .Producing nanofibers with excellent physico-biochemical characteristics using an electrostatic spinning technique is considered to be an efficient multipurpose drug delivery strategy for delivering various therapeutic agents ranging from nanomedicines to macromolecules, including proteins and nucleic acids. Nanofibers can overcome several key challenges such as low solubility, loading efficiency, short circulation and the plasma half-life of drugs and are effective at improving the bioavailability of drug growth factors, DNA, RNA, and so forth. . Using vDiscovering a new potential treatment strategy against cancer is very difficult due to several drawbacks of anticancer therapeutics such as imperfect solubility, impermanence in the circulatory bloodstream, poor accumulation in cancer cells, highly toxic to normal cells, low working efficiency in solid tumors and excess elimination profile . To surpRecently, electrospun nanofibers have been widely used to ameliorate the solubility of anticancer drugs a. In thiEven though polymeric nanoparticles (NPs) are used as a drug carrier to load anticancer drugs like Paclitaxel, the poor drug-loading efficiency of these systems minimizes the applications in the biological system . On the DOX, 500 nm) and these nanocomposites were injected into a tumor-bearing mice model, it showed 4 times more accumulation in the cancer site and was 3 times more stable in plasma level compared with microspheres. However, small length nanofibers of 2 micrometers in diameter exhibited the most powerful cancer cell apoptosis and necrosis activities with the highest resistance to metastasis -2-methoxybenzamide) is an antidopaminergic benzamide, pharmacologically used as a serotonin receptor agonist. Its inhibitory action on acetylcholinesterase exerts its prokinetic and anti-emetic effect due to its actions on contractility of colonic smooth muscle. Recently, Jaber and co-workers fabricated a core/shell nanofiber using PVA/PCL to load metoclopramide hydrochloride [Metoclopramide  as well as core-shell nanofibers such as PVA/PCL, PVA/PLLA, and PVA/PLGA to investigate the control release behavior of metoclopramide loaded as-synthesized nanofibers. The drug release profile data suggested that the controlled release of hydrophilic entities is possible by using core/shell nanofibers and by verifying the physicochemical properties of core/shell solutions. The result also showed a clear difference according to the release characteristics between the monolithic fibers, which are made of hydrophilic and hydrophobic polymers, and core/shell fibers using PCL, PLLA, and PLGA 80/20 shell polymers. The monolithic fibers cannot control the initial burst release of the hydrophilic drug metoclopramide hydrochloride but core/shell electrospun nanofibers can easily regulate the controlled release of incorporated drugs. Thus, electrospun nanofibers would be a promising gastrointestinal drug carrier to achieve controlled release behavior and protect sensitive drugs in a biological pH .Antihistamine generally works by blocking the physiological activities of histamine, thus, antihistamines are used in the treatment of nasal congestion, sneezing, hives, seasonal hay fever and especially, to relieve the symptoms of various allergies such as dust allergy, cold allergy, allergic rhinitis, indoor and food allergies. For the time being, electrostatic nanofiber scaffolds are being used as a carrier of various antihistamine drugs. One experimental study was designed to incorporate the first-generation antihistamine (H1 receptor antagonist) chlorpheniramine maleate into glutinous rice starch combining polyvinyl alcohol (GRS/PVA) electrospun nanofibers to investigate a drug delivery carrier concept and control release properties of the nanofibers . The hybDevelopment of fast dissolving delivery systems of therapeutics (FDDST) is an excellent and unique concept especially for those patients who have difficulties swallowing pharmaceutical solid dosage forms. FDDST offers very fast-dissolving solid oral dosage forms that take a few minutes for absorption in the patient’s mouth. Thus, FDDST facilitates high drug bioavailability, therapeutic window and exemption of hepatic first-pass elimination. Previously, the electrospinning method was employed to design, develop and evaluate FDDST in which one study proved more than 80% of total ibuprofen release within 20 s  whereas Another example of an antihistamine is Diphenhydramine, a member of the ethanolamine class of histamine H1 receptor antagonists, which is mainly used to treat allergies, nausea, motion sickness, extrapyramidal symptoms and symptoms of a common cold by reversing the effects of histamine on capillaries. It is also used in parkinsonism due to its ability to act as a muscarinic acetylcholine receptor reverse agonist by crossing the blood-brain barrier. In addition, its usage as a sodium channel blocker introduced it as a local anesthetic . HoweverFor the first time, a composite electrospun nanofiber was fabricated by free-surface electrospinning with various microscale geometries as a carrier of physicochemical diverse medicines including the contraceptive drug progestin levonorgestrel . The as-Palliative treatment—also known as comfort or supportive care—aimed to relieve, reduce or control symptoms, side effects and adverse effects such as pain and sickness at any stage of serious diseases, thus enhancing the life expectancy and comfort of a patient even during the last stages of an illness. Nagy et al. used lower molecular weight poly to fabricate the non-woven tissues of a nanofiber with a large surface area for developing an oral fast-dissolving dosage form by applying electrostatic spinning. After loading highly water-soluble Donepezil HCl (which is used to treat dementia-related Alzheimer’s disease) into as-prepared nanofibers, the diameter of the electrospun nanofiber was 100–300 nm even when the polymer-drug ratio was 33 w/w%. Interestingly, in comparison with the release rate of the commercially available Aricept tablet (brand name of Donepezil HCl), the drug-loaded nanofiber took less than 30 s to release whereas the Aricept tablet took ≥30 min. This might be due to the availability of the nanofiber’s large surface area which is directly proportional to the dissolution rate, corresponding to the Noyes and Whitney equation [Among various miscellaneous drugs, caffeine is the most used agent worldwide that binds to adenosine receptors by mimicking natural adenosine, affecting various body functions. At present, electrospinning techniques are being widely used to fabricate caffeine loaded nanofibers as a FDDST facilitating a high surface area and high porosity of nanofibers that can lead to fast-wetting features of the nanofiber surface to ensure quick release properties of incorporated drugs. In these respects, drugs are usually encapsulated into nanofibers either in amorphous or nanocrystal format, ensuring high solubility, dissolution, rapid onset of action and bioavailability of loaded drugs. A study was designed by electrospinning using PVA as the drug nanocarriers of caffeine (CA) and riboflavin (RFN) in which the dissolution time and wetting time were 1.5 s and 4.5 s respectively for both PVA-CA and PVA-RFN. In addition, the release percentages of CA and RFN from PVA filament-forming nanofibers were 100 and 40 respectively within 1 min . In anotDiscovering an effective and potential ophthalmic drug delivery system (ODDS) is still a challenge due to the quick elimination of ocular drugs like eye drops. Thus, rapid peroneal losses of eye drops, even with the high volume of administration resulting in low bioavailability and very short half-life of applied drugs and a lower amount of ophthalmic drugs , can reach into intraocular tissues via penetration through the cornea. To address these problems, Gagandeep et al. (2014), developed a nanopatch using PVA and PCL via electrostatic spinning to load and deliver a combination of timolol maleate and dorzolamide hydrochloride as a model drug against glaucoma. As-developed drug-loaded nanopatches were 200–400 d·nm with almost 100% drug entrapment efficiency—no interaction has been found between polymers and encapsulated drugs. Furthermore, the in vitro drug release experiment alluded an initial burst release of incorporated drug from nanopatches followed by controlled release behavior for up to 24 h . The oveDrug enrichment on the surface of the polymer is only possible if the drug is blended with the polymer solution before electrostatic spinning. This system facilitates an initial burst release of the loaded-drug resulting in rapid onset of action, but it also decreases the potential lifetime of delivery carriers. Hence, to obtain desired results such as sustained therapy, local and controlled release, enhanced activity and retrenchment of side and adverse effects, core-shell nanofibers would be the best option. One study was designed to load brimonidine tartrate (BT) into a first dissolving dendrimer nanofibers (DNF) against glaucoma a. The inThe delivery of nucleic acid molecules such as DNA or interfering RNA, into a target cell to knockout/ knockdown mutated gene expressions, either by gene editing or impeding the mechanism of the mutated gene, has proven its novelty in the field of regenerative disease treatment. In this case, when a gene misbehaves in a protein, gene therapy is able to introduce the new copy of that specific gene for recuperating the function of that protein by adjusting the signal transduction pathway . To dated,l-lactide)–poly(ethylene glycol) type biodegradable synthetic copolymers [However, the encapsulation of a nucleic acid/polymer complex in the core of core/shell nanofiber scaffolds can protect the encapsulated DNA or interfering RNA from biological degradation and denaturation, as well as prolong their release up to several months by controlling it. For instance, non-knitted, membraneous nanofiber scaffolds were fabricated via the electrospinning procedure as a gene delivery carrier, containing plasmid DNA encapsulated with poly(lactide-co-glycolide) and poly), PLGA (poly(lactide-co-glycolide)), LPEI (linear polyethyleneimine), ELP (elastin-like polypeptides), PDLLA ), and many more are used as a nucleic acid carrier [However, the hydrophobic nature and slow degradation properties of PCL are responsible for the slow release properties of encapsulated siRNA ,121. To  carrier ,123,125 Growth factors are naturally occurring endogenous signaling molecules  which are efficient for inciting cellular development, differentiation, regeneration and proliferation by binding to cellular receptors. Recently, marked progress has been made in preparing several types of ultrathin electrospun nanofiber scaffolds to successfully load and deliver several growth factors a as wellThe quick inactivation and very short biological half-life of growth factors are the most important factors hindering their effectual delivery. In addition, it has been noted that the growth factors’ carrier should have slow and sequential releasing properties. The establishment of growth factors into nanofiber scaffolds play very crucial roles in repairing and regenerating damaged tissues by mimicking signal transduction from cell to cell, or from cell to its extracellular matrix . Either Recently, the polysaccharide matrix is being used as a superior nanofiber material to fabricate a growth factor carrier owing to their low toxicity, biocompatibility, and extracellular matrix mimicking capability. However, introducing the sulfation state to the alginate scaffolds ensures the appointed binding and loading of growth factor into the polymer backbone, as well as maintenance of its control release properties. For instance, Mohammadi S, et al. fabricated electrospun scaffolds by blending alginate sulfate and polyvinyl alcohol to load and deliver transforming growth factor-beta1 (TGF-β1) for tissue engineering applications. For the first time, this research revealed that more successful binding, encapsulation and sustain release of heparin-like proteins from electrospun scaffolds could be achieved by the addition of an affinity site to the alginate . A part Usually, the traditional nanofiber scaffolds-based GFs delivery methods such as dip coating and covalent binding are associated with low entrapment efficiency and primary burst release properties. Currently, researchers are looking for new, innovative and stable strategies by focusing on controlled and sustained release properties. To control the release properties of encapsulated GFs, the usage of gelatin particles in nanofiber fabrication is becoming popular. Generally, the in vitro degradation time of gelatin particles is about 240 h, so researchers tend to use gelatin particle-mediated nanofiber scaffolds as a promising GF nanocarrier. The basic advantages of using gelatin particles are their slow degradation and controlled release properties in a biological environment because of introducing cross-linking treatment during fabrication. An example would be the work of Jiang and co-workers who prepared PCL fibrous scaffolds containing vascular endothelial growth factor (VEGF)-incorporated gelatin particles which were able to exhibit both diffusion and degradation mediated release for a long period of time b, differd,l-lactic acid) (PDLLA) nanofiber scaffolds whereas the glial cell line-derived neurotrophic factor was incorporated in poly(lactic-co-glycolic acid) (PLGA) electrospun scaffolds [Biocompatible and biodegradable polymeric nanoparticles and biomimetic hydrogel have gained more attention for encapsulating various types of growth factors and confirming their controlled and prolonged releasing . VEGF ancaffolds . This ducaffolds . The prel-lactide-co-caprolactone) and poly(2-ethyl-2-oxazoline) was able to protect the bioactivity of dual angiogenic basic fibroblast growth factor and vascular endothelial growth factor during and after their physical blending. It is also capable of mimicking the topography and chemical cues of native cardiac tissue structures [d,l-lactic acid) [The time period of sustained release of GFs from its carrier should be proportional to the time period of GFs capability to preserve the nanofiber scaffolds; therefore, this important factor is considered during the design of nanofiber-mediated GFs delivery. Employing several electrospinning techniques, the preservation of GFs’ bioactivity is achieved for the proliferation and migration of specific cells. The heterogeneous electrospun matrix made of biocompatible poly(ructures . The strructures . The priructures . Previouic acid) . This glElectrospinning is a promising technology for manufacturing nanofibers from the laboratory to industrial level. Nanomaterials possess a large surface area and enhanced porosity, which are advantages for drug delivery and many other biomedical applications. Nanofibers can be formed using three different techniques: electrospinning, self-assembly, and phase separation. Electrospinning is the most widely used technique because of its promising results. Electrospun nanofibers have attracted much attention due to their biocompatibility, adhesiveness, sterile nature and their efficiency in diverse applications. Currently, nanofibers are considered suitable candidates for scaffold materials, wound dressing materials, drug delivery systems, filtration membranes, catalysts for reduction and so on. Nanofibers are used for medical applications, batteries and fuel cells as a new material.Though the application of nanofibers is versatile, still more development and research is required for the proper use of nanofibers, especially for biomedical therapeutic delivery. The application of nanofibers in drug delivery, medical implants or tissue engineering is still very limited and most of the materials cannot reach the stage of clinical trials. A thorough investigation on the scaling up of nanofiber technology is required, to make it widely available.


Rewarding stimuli  induced an increase in dopamine efflux in the NAc of wild-type (WT) mice. The dopamine responses were attenuated (cocaine) or abolished  in constitutive p11 knock-out (KO) mice. The dopamine response to cocaine was accompanied by an increase in ACh NAc efflux, whereas the attenuated dopamine response to cocaine in p11 KO mice was restored by activation of nicotinic or muscarinic ACh receptors in the NAc. Dopamine responses to rewarding stimuli and ACh release in the NAc were attenuated in mice with deletion of p11 from cholinergic neurons (ChAT-p11 cKO mice), whereas gene delivery of p11 to CINs restored the dopamine responses. Furthermore, chemogenetic studies revealed that p11 is required for activation of CINs in response to rewarding stimuli. Thus, p11 in NAc CINs plays a critical role in activating these neurons to mediate dopamine responses to rewarding stimuli. The dysregulation of mesolimbic dopamine system by dysfunction of p11 in NAc CINs may be involved in pathogenesis of depressive states.A recent study showed that p11 expressed in cholinergic interneurons (CINs) of the nucleus accumbens (NAc) is a key regulator of depression-like behaviors. Dopaminergic neurons projecting to the NAc are responsible for reward-related behaviors, and their function is impaired in depression. The present study investigated the role of p11 in NAc CINs in dopamine responses to rewarding stimuli. The extracellular dopamine and acetylcholine (ACh) levels in the NAc were determined in freely moving male mice using  The reduction of p11 expression in NAc CINs is tightly associated with anhedonia as well as other depression-like symptoms of behavioral despair. To improve therapeutic efficacy of antidepressants for anhedonia, a new type of antidepressant directly or indirectly acting on the mesolimbic dopamine pathway needs to be developed. For this purpose, therapeutic strategies that increase the function of p11 and its signaling pathway in NAc CINs may have an impact on antidepressant efficacy.1B and 5-HT4 receptors at the cell surface  and decreased positive affect  in the NAc and PFC were similar between wild-type (WT) and constitutive p11 KO (p11 KO) mice  mice . CocaineKO) mice C. ExposuKO) mice D,E. The KO) mice F–H. Thesp11 is highly expressed in NAc CINs  and is i-/- p11flox/flox mice; + p11flox/flox mice). These results indicate that p11 in ChAT cells plays a critical role in the dopamine responses to rewarding stimuli.To directly investigate the role of p11 in ChAT-expressing cells, the dopamine responses to rewarding stimuli were evaluated in the NAc of ChAT cell-specific p11 KO mice (ChAT-p11 cKO mice), which were obtained by mating p11 floxed mice with ChAT-Cre mice . The basAAV-loxP-RFP/stop-loxP-p11 (AAV-p11) in the NAc of ChAT-p11 cKO mice (AAV-p11) into the NAc induced the expression of RFP in ChAT-Cre-/- cells such as medium-sized spiny neurons and GABAergic interneurons  was injected into the NAc. YFP expression induced by ChAT-Cre was indeed observed in RFP-negative ChAT expressing cells -mCherry) or control (AAV-DIO-mCherry) virus was injected into the NAc of ChAT-p11 cKO mice. After four weeks of Gs-DREADD viral injection, mCherry was expressed in ChAT-positive large-sized neurons in the NAc  for 140 min did not affect the basal levels of dopamine, cocaine infusion (1 µM) into NAc was started. Cocaine infusion together with CNO infusion (3 µM) induced the dopamine responses in the NAc of ChAT-p11 cKO mice with Gs-DREADD viral injection C. RestorAAV-DIO-rM4D(Gi)-mCherry) or control virus (AAV-DIO-mCherry) (We further investigated whether the inhibition of NAc CINs by Gi-DREADD could suppress the dopamine response to cocaine infusion in the NAc of ChAT-Cre mice injected with Gi-DREADD virus (mCherry) . In ChATIn vivo microdialysis analyses in constitutive p11 KO mice revealed that lack of p11 induced the attenuation of dopamine responses to rewarding stimuli including a drug of abuse and natural rewards. The attenuation of the dopamine responses in the mesolimbic (VTA-NAc) dopamine system, but not in the mesocortical (VTA-PFC) dopamine system, suggested the importance of p11 in the NAc. The dopamine responses were attenuated in ChAT-p11 cKO mice, and the attenuated responses were restored by the overexpression of p11 in NAc CINs, indicating the critical role of p11 in NAc CINs. Furthermore, lack of p11 in NAc CINs results in the attenuation of ACh release in response to cocaine and the subsequent decrease in nicotinic and muscarinic ACh receptor signaling at dopaminergic terminals, leading to the suppressed dopamine responses to cocaine and possibly other rewarding stimuli. The function of p11 in CINs was confirmed by the chemogenetic studies: CIN activation by Gs-DREADD restored the dopamine responses in ChAT-p11 cKO mice, whereas CIN inhibition by Gi-DREADD suppressed the dopamine response in control (ChAT-Cre) mice. Thus, p11 in NAc CINs is required for the dopamine response of the mesolimbic rewarding system. These findings provide insights into the neural mechanisms of anhedonia in depression.In this study, we demonstrated that p11 expressed in CINs of the NAc is a critical regulator of the dopamine reward system. p11 regulates the dopamine response to rewarding stimuli in the mesolimbic dopamine pathway, but not in the mesocortical dopamine pathway. Selective regulation of the mesolimbic dopamine pathway is enabled by action of p11 in CINs of the NAc. PFC receives cholinergic innervation from the basal forebrain , and ChA1B receptor, 5-HT4 receptor and mGluR5 are required for antidepressant action (1B receptors in CINs may induce the inhibition of CIN activity (Cholinergic tone in the mesolimbic dopamine system plays an important role in behavioral responses to psychostimulants and natural reward . It has t action , but theactivity , but thiAnhedonia is a core symptom of depression. It has been shown that anhedonia is associated with a deficit in the mesolimbic dopamine circuit . CurrentDeletion of p11 in ChAT cells, p11 knock-down in the NAc or silencing NAc CINs induces anhedonic behavior, and overexpression of p11 in NAc CINs reverses anhedonic behavior in constitutive p11 KO mice . In addiIn conclusion, p11 is a critical regulator of CIN activity as measured by the dopamine response of the mesolimbic dopamine pathway to rewarding stimuli. p11 is required for reward-mediated NAc CIN activation and induction in ACh release, resulting in the enhancement of dopamine release. To improve therapeutic efficacy of antidepressants for anhedonia, a new type of antidepressant directly or indirectly acting on the mesolimbic dopamine pathway needs to be developed. For this purpose, p11 and its complex in the NAc CINs may be good therapeutic targets.