As most resins had not been hydrolysed, fibre extraction was difficult. After 21–25 days of retting, the tensile strength of fibres was about 34 ± 10 cN·tex−1, elongation at break was about 1.71%, and moisture regain was about 13.56%. The fibre characteristics met the process requirements, and the tensile separation stress of the stem was small, about 0.034 MPa. This time point could be used as the optimum endpoint for retting flax in warm water, which could provide theoretical support and research basis for the recycling of banana straw. The functional groups of the extracted fibres were studied by FTIR, which confirmed the observed change rule of each component during degumming. The experimental results showed that a longer retting time corresponded with a lower content of fibre impurities, more thorough degumming, and less difficult extraction; however, strength and toughness decreased.In this paper, warm-water flax retting was used as a pretreatment method for banana-fibre extraction. To determine the optimum conditions for flax retting, the physical properties of various parts of stems and fibres in the process of flax retting were analysed. By studying the tensile strength, elongation at break, diameter, moisture regain, and other characteristics of the fibres, the influences of bacteria and enzymes in the retting liquor on the fibre characteristics in different retting stages were determined. Through mechanical-property tests and microscopic observation of the stem skin, the change rules of the mechanical properties and degumming state of the stems were examined. The results showed that the fibre tensile strength of banana stems reached the maximum value of 45 ± 16 cN·tex Given the increasing awareness of the global energy crisis and environmental problems, the development and research of plant fibres are attracting considerable attention . Plant fBanana fibre is a plant fibre extracted from banana-stem waste. Its light, soft, and easy-to-weave nature confers good market advantages and development potential . In 2021The extraction methods of banana fibre include the mechanical separation of non-cellulose components and the chemical or microbial dissolution and decomposition of such components . MechaniIn summary, extracting banana-stem fibre by the biological method is feasible, but it is hounded by some problems, such as high cost, low fibre-extraction efficiency, and difficult observation of the fibre-extraction process. The method of retting flax in warm water is a classical biological method. It uses the enzymes and bacteria in the banana stem to provide an environment suitable for its reproduction and then completes the fibre extraction. Under the action of the complex flora in the retting pool, the non-cellulose components in the stems are decomposed, and the fibre bundles are gradually separated from the plant tissues, thereby facilitating the extraction of the fibres . The purThe banana stems used were from ‘Brazil Williams’ bananas planted by Danzhou agricultural community. The water source for retting flax was local tap water.The banana stems were pretreated with warm water retting. Fibre is distributed in different parts of a banana tree, so the mechanical properties of fibre significantly differ . As showAccording to the ASTM D3379-75 standard, the tensile strength and elongation at break of fibre bundles were measured using a tensile tester . The banana stems were stripped and sampled manually every other day from the fifth day of retting. The sampling parts were evenly distributed throughout the entire stem slice. Twenty fibre samples were taken from each stem slice to obtain banana stem fibre bundle samples with a length of more than 40 mm. The fibre bundles were clamped between two pneumatic clamps with a test length of 25 mm, and the traveling speed of the pneumatic clamp was set to 2 mm/min.Ten fibres were randomly taken from each stem slice, and six different positions were observed for each fiber. The diameter of fibres was measured with a polarised light microscope  at 100× magnification, and the average value and coefficient of variation were calculated.The determination of moisture regain of banana-stem fibres was in accordance with the GB6503-2008 standard, using a dryer  and a one ten-thousandth balance (DEJUINSPR 124ZH/E). Five samples with a length of 40 mm for each group, were weighed, dried at 105 °C for 2 h, and the dried mass then weighed. Three groups of tests were conducted for each batch of samples, and the moisture regain of the samples and the average value were calculated.max is ultimate tension, and S is fracture section area.The retting time and different parts of the sample are the main factors affecting the tensile separation stress of a fibre bundle. From the beginning of retting, the banana stem was sampled every other day. A blade was used to separate the outer skin of the banana stem, and the sample was cut into rectangular samples with a length of a = 60 mm and a width of b = 30 mm. As shown in This paper mainly used the descriptive statistical analysis method. The specific data statistical analysis process was as follows. In the mechanical property analysis experiment, the data corresponding to the mechanical characteristic curve of the sample was imported into Microsoft Office Excel 2013 software, and the tensile starting point (the first data point with a load greater than 0.2 N) and the maximum load point were selected. The data was imported into Origin 2017 software to draw the statistical graph. In the determination of other physical properties, the data measured in the test were counted with Statistical Product and Service Solutions 19.0 software to complete the calculation of standard deviation, average value and coefficient of variation.The surface of the outer epidermis of the stem slice was observed under an inverted microscope , and the magnification was 40 times.−1, and the resolution was 2 cm−1. The sample was scanned 32 times, and the functional image with wavenumber as independent variable was the output.The change in functional groups of the samples could be evaluated by FTIR spectroscopy. The instrument used was a near-infrared spectrometer . Initially, samples of banana-stem fibres that had been retted for 5, 15, and 25 days were initially ground and mixed with KBr. The mixture was ground in an agate mortar and compressed into discs. The spectral recording range of the sample was 600–4000 cm−1 for the entire retting process at about 11 days (about 15% higher than that of untreated fibres). The changes in this process included the decomposition of some pectin in the fibres (primarily due to the role of anaerobic pectin-decomposing bacteria) and the direct or indirect dissolution of lignin and its hemicellulose connected with cellulose by the action of enzymes [−1. It is difficult for the lower stem to reach this strength standard, and so it is often not used as the raw material for fiber extraction in production [ enzymes . After r enzymes . In turn enzymes . After 3oduction . In compBy comparing the tensile-strength changes of the fibres in the upper, middle, and lower parts of each layer of culm, we found that the strength of banana stem fiber has the same distribution law as hemp fiber, that is, the lower fiber has the lowest strength and the middle fiber has the highest strength. The reason for this is that the lower fiber diameter is significantly higher than the upper and middle fiber diameters. The larger the fiber diameter, the higher the probability of containing defects. The difference between the middle fiber diameter and the upper fiber diameter is small, but the proportion of cellulose in the middle fiber is higher than the upper fiber, so the fiber strength is higher . Under tThe tensile mechanical properties of fibre bundles are scattered, affected by various parameters, such as sampling position, growth characteristics, and degumming . ThroughThe tensile mechanical properties of fibres are the most important physical properties of fibres. In order to obtain high-quality fibres, the higher the tensile strength of fibres, the better, and the smaller the discrete type of fiber tensile strength, the better. It is comprehensively considered that 21–25 days is the best time end point for retting.As shown in From the standard deviation of the moisture regain of banana stem fibre, we found that the moisture regain was a very stable physical property of banana fiber. Its standard deviation was always lower than 5% of the average value, and this balanced property was not affected by the degumming in warm water. This property means banana fibres have more application scenarios .The microscopic images of the outer skin of banana-stem slices after 5, 15, and 25 days of retting are shown in −1 could be reached, corresponding with OH stretching vibration [−1 corresponded with the CH tensile vibration of CH and CH2 in cellulose and hemicellulose, respectively [−1 corresponded with the CH tensile vibration in hemicellulose [−1 corresponded with the C=C tensile vibration of aromatic group [−1 corresponded with the bending vibration of lignin CH2 [−1 corresponded with the C-O tensile vibration of the acetyl group or the C-O-C tensile vibration of the ester group [−1 corresponded with the glucocyclic β-glycosidic bond [ibration . The twoectively . The peaellulose , that atic group , that atgnin CH2 , that ater group , and thadic bond .−1, the peak value gradually increased as retting proceeded, indicating that hemicellulose and lignin continuously hydrolysed during retting. With the gradual weakening of the intensity of the peak in the process of retting at 1734 and 1256 cm−1, the content of hemicellulose gradually decreased, the peak at 1734 cm−1 in curve c was missing, and the peak at 1256 cm−1 significantly weakened, indicating that the content of hemicellulose in banana fibres was very small after 25 days of retting. With the treatment of fibres at 1516 and 1256 cm−1, the peak gradually weakened because of the hydrolysis and dissolution of lignin, and the lignin content in the fibres decreased. The characteristic peak at 898 cm−1 existed in all curves, corresponding with the β-glycoside bond representing cellulose, indicating that cellulose was well retained throughput the entire retting process.Through FTIR analysis, we concluded that no new functional groups were introduced into the fibres during retting, so no other impurities were introduced into the retting process. By comparing curves, a, b, and c, in −1, was reached after 11 days of retting, and was lower than the tensile strength standard of high-quality fiber after 25 days of retting, and reached the minimum tensile strength of 19 ± 5 cN·tex−1 after 31 days of retting. The elongation at break of the fibres remained unchanged at first and then decreased. At 9 days after warm water retting, it remained at 2.1 ± 0.2% and then decreased to 0.9 ± 0.1%. The fiber diameter decreased from 240 ± 20 μm to 187 ± 7 μm. The moisture absorption of the fiber was usually reflected by the moisture return rate, which decreased from 15.0 ± 0.9% to 12.6 ± 0.8%. The above physical properties became concentrated. A longer retting time corresponded with a lower content of fibre impurities, more thorough degumming, and less difficult extraction; however, strength and toughness decreased. According to comprehensive analysis, in order to obtain high-quality fibres with tensile strength up to standard and concentrated physical properties under the condition of low fiber extraction difficulty, the end time of retting should be about 21–25 days. Notably, the lower stem carried more enzymes and bacteria required for retting, so the biological reaction process was more rapid and thorough. If centralised retting was performed, retting should be properly terminated ahead of time based on the time endpoint determined in this paper. Finally, the removal of hemicellulose and lignin and the retention of cellulose were confirmed by FTIR spectroscopy analysis of banana fibres. No chemical reagents were used throughout the entire retting process, so the cellulose was not seriously damaged. The retting process was environmentally friendly, and almost no pollutants were generated.Retting flax in warm water under natural conditions was used to pretreat banana stems, and the effects of retting treatment on the physical and chemical properties of banana stems and banana fibres from different parts were studied. Bacteria, such as pectin-decomposing bacteria, and enzymes, such as xylanase and pectinase existing in the banana stem, multiplied in the retting pool. These enzymes and bacteria changed the chemical composition of fibres, and the content of hemicellulose, lignin, and pectin gradually decreased, and the mechanical properties, diameter and moisture absorption of the fibers changed. Experimental results showed that the tensile strength of the fibers increased slowly at first and then decreased rapidly with the retting process. The maximum tensile strength, 45 ± 16 cN·tex


Vibrio vulnificus is typically present in seawater, fish, and shellfish, and is known to cause severe sepsis, particularly in patients with liver diseases such as cirrhosis. V. vulnificus is one of the most dangerous waterborne pathogens, and infection mainly occurs in western Japan during the summer, with an increased fatality rate. Herein, we report the case of a patient with primary biliary cholangitis and sepsis caused by V. vulnificus infection sustained through shrimp shelling.V. vulnificus; thus, minocycline was administered in addition to meropenem. The disease course was uneventful, and the patient was discharged on day 28 of hospitalization. Symptoms in the right upper arm developed 1 day after the patient shelled a large number of shrimp; therefore, the infection route was assumed to be through wounds sustained during shrimp shelling. We suspected liver disease and measured serum anti-mitochondrial M2 antibody levels, leading to the diagnosis of primary biliary cholangitis.An 82-year-old Japanese Asian woman with no medical history or underlying disease developed redness, swelling, and pain, which extended from the right fingers to the upper arm. A diagnosis of sepsis due to cellulitis was made. Blood culture detected V. vulnificus infection should be thoroughly assessed for the presence of liver diseases such as primary biliary cholangitis.As in this case, small wounds caused by handling fish and shrimp are a potential source of infection. Patients with severe  Vibrio vulnificus infection were first reported in 1979 by Blake et al. V. vulnificus is a slightly halophilic Gram-negative bacillus with an optimal salt concentration of 2–3% that lives in seawater and brackish water [V. vulnificus infections mainly occur in western Japan during the summer, and it is considered one of the most dangerous waterborne pathogens, with a reported fatality rate of 50% [V. vulnificus, exposure to seawater, or wounds inflicted by marine animals. V. vulnificus infection can result in fatal sepsis in patients with underlying diseases, such as preexisting liver disease, hemochromatosis, or immunodeficiency. To the best of our knowledge, there are no case reports of primary biliary cholangitis (PBC) associated with V. vulnificus infection. Herein, we report the case of a patient with PBC and sepsis caused by V. vulnificus infection sustained through shrimp shelling.The clinical and epidemiological characteristics of The patient was an 82-year-old Japanese Asian woman with no comorbidities, relevant family medical history, alcohol consumption, or known allergies. No previous liver disease or abnormal liver function tests were noted. However, she had been a smoker of 20 cigarettes per day until 3 years prior. After mountain climbing on 23 and 24 July 2012, she experienced numbness and pain in the right arm. On 25 July, she developed general malaise. Since her symptoms did not improve by the morning of 26 July, she visited a private hospital with facial pallor, poor dietary intake, redness, mild swelling, and pain in the right arm. She was in circulatory shock with a blood pressure of 60/48 mmHg, and was later transported by an ambulance to Fukuoka University Hospital.2; conscious and coherent; blood pressure, 66/33 mmHg; pulse rate, 91 beats per minute ; respiratory rate, 22 breaths/minute; SpO2, 97% (room air); body temperature, 37.7 °C; and redness and swelling extending from the medial part of the right upper arm to the right elbow and forearm  twice daily for 8 days, to which V. vulnificus was susceptible  for 12 days, with linezolid (1.2 g/day) for 5 days and immunoglobulin (5 g/day) for 3 days via intravenous injection. On day 2 of hospitalization, a blood culture detected Gram-negative bacilli , as the redness and swelling in the upper arm had improved. After 7 days, drug-induced thrombocytopenia was observed, and levofloxacin was discontinued. No recurrence was observed, and the patient was discharged on day 28 of hospitalization Fig. .Fig. 4DiDuring the course of hospitalization, persistent elevation of serum γ-glutamyl transpeptidase, alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase levels suggested liver disease. Blood tests performed on day 5 of hospitalization showed positivity for anti-mitochondrial M2 antibodies , particularly those with underlying diseases, such as liver disease, diabetes, immune disorders, elevated serum iron concentration, and cirrhosis  [V. vulnificus infection is characterized by a short incubation period, typically within 24 hours of exposure. Moreover, since this bacterium can cause severe infections, such as bacteremia and wound infections, prompt initiation of antibacterial therapy is essential. Therefore, if an infection is suspected, particularly in high-risk patients, it is crucial to identify this bacterium accurately and rapidly in clinical practice [V. vulnificus caused wound infections and bacteremia in 62 patients over the course of 1 year in Israel. Among them, 57 patients developed cellulitis, four developed necrotizing fasciitis, and one developed osteomyelitis. The fatality rate of V. vulnificus infection has been reported to be 20–50%, and no deaths were observed in this particular report [induced) . V. vulnV. vulnificus is influenced by water temperature. V. vulnificus accounted for approximately 8% of the aerobic bacteria in samples collected from the Chesapeake Bay between April 1991 and December 1992. It was not detected in February or March (water temperature < 8 °C) but was detected in 80% of the samples in May, July, September, and December (water temperature > 8 °C) [V. vulnificus became temporarily undetectable in this area. Therefore, salt concentration greatly influences the growth of V. vulnificus [V. vulnificus.The proliferation of  > 8 °C) . Regardilnificus . In the et al. in 1857; however, its exact cause remains unknown [Escherichia coli can disrupt immune tolerance, leading to the development of PBC [V. vulnificus infection occurred after the patient originally had PBC, or that PBC developed after V. vulnificus infection. It is also possible that the two diseases coincidentally overlap. However, there are no precedents for either of these possibilities, and we cannot discuss these possibilities at this time.PBC, which is an autoimmune disease associated with environmental and genetic factors, was first reported by Addison  unknown . A familt of PBC , 7. In tV. vulnificus infection for the presence of undiagnosed liver disease, such as asymptomatic PBC. In addition, a detailed history regarding V. vulnificus infection should be obtained because shelling of crustaceans is a potential source of infection.In conclusion, the lessons learned from this case suggest that it is useful to evaluate patients with


Klinefelter syndrome (KS) is a common cause of non-obstructive azoospermia (NOA). Advances in fertility preservation (FP) techniques, such as the use of microdissection testicular sperm extraction (micro-TESE), have improved sperm retrieval rates (SRR) up to 40–50% in this population. Age has been suggested to have an impact on FP, postulating that sperm production may deteriorate over time due to germ cell loss. As such, sperm retrieval for patients with KS at a younger age has been proposed to further improve SRR; however, whether such practice pragmatically improves SRR is yet to be determined, and controversy remains with concerns over trauma caused by FP procedures on further impairment of testicular function. There has also been a debate on the ethics of performing FP procedures in the pediatric population. Optimizing FP for patients with KS invariably requires a holistic multidisciplinary approach. This review aimed to evaluate the latest evidence in performing FP in pediatric patients with KS, and discuss the controversy surrounding such practice. Hormonal changes in patients with KS during childhood and the use of hormonal manipulation to optimize SSR in this population have also been reviewed. Klinefelter Syndrome (KS) is a chromosomal disorder in which there is at least one extra X chromosome when compared with the normal male karyotype of 46, XY. It has an estimated prevalence of 1 in 500–600 males, with approximately two-thirds diagnosed in adulthood at a mean age of 27.5 years , accounting for ~10% of all patients with azoospermia . As suchThe current focus of pediatric KS treatment surrounds targeting endocrine deficits to improve growth, muscle mass, and prevent delayed puberty. Recent investigations in patients with KS and NOA have demonstrated a time-sensitive pattern of germ cell depletion, suggesting younger age is a potential positive predictor of success in sperm retrieval . The rolThe endocrinology of men with KS is typically recognized by a hormone profile of high serum follicle-stimulating hormone (FSH), low luteinizing hormone (LH), and low testosterone levels; however, various significant changes occur from birth to adulthood that impacts spermatogenesis in patients with KS.During early development in males with KS, the first peak of testosterone occurs at “mini-puberty,” which involves the activation of the hypothalamus-pituitary-gonadal axis within the first 3–6 months of life. During this time, testosterone, FSH, and LH levels can reach pubertal or even adult levels; however, there is contrasting evidence on whether testosterone levels are comparatively lower inpatients with KS during mini-puberty to healthy males, suggesting a role of Leydig cell dysfunction occurring early in childhood , but a rin-situ hybridization (FISH) study in 10 patients with KS   with KS . Surgica with KS –18.n =11) (n = 2) (n = 4)  , in 4.2% (n = 2) , in 7.7% (n = 4) , and in  (n = 4) , 23. The (n = 4)  of eightSurgical retrieval remains the most utilized and reliable method of retrieving sperm in patients with KS.Conventional testicular sperm extraction (cTESE) was used prior to the late 1990s for harvesting sperm in patients with KS, where testicular parenchyma is biopsied through incisions in the tunica albuginea to obtain random seminiferous tubule specimens to search for sperm; however, this technique was disappointing, with reported SRR of only 25 to 40% . Postulavia TESE in a pediatric setting was first reported in 2001 by Damani et al. (p < 0.01). Although these results appeared to be encouraging, available literature investigating FP in young patients with KS has limited sample sizes, and the reported wide range of SRRs is likely due to variation in testosterone pre-treatment, mosaicism/non-mosaicism, and level of testicular dysfunction. As such, no current guidelines exist on optimal timing for FP in KS.Sperm retrieval i et al. , who suci et al. . Unlike i et al. . Early Fi et al. . This isi et al. . Bryson i et al.  conducteOn the contrary, several studies showed low or comparable SRRs in young men with KS. Plotton et al.  used spein vitro spermatogonial stem cell (SCC) propagation and auto-transplantation has been explored recently to provide another means of treating patients with KS and NOA. A meta-analysis  in response to LH are critical for spermatogenesis. Normally, endogenous testosterone directly inhibits GnRH and LH release at the hypothalamus and pituitary levels, leading to downstream attenuation of testosterone production ; howeverTestosterone supports the development of secondary sexual characteristics, adequate bone mass, and improved muscle strength. Furthermore, testosterone therapy has been shown to have an important role in reducing the risk of cardiovascular diseases and metabolic syndrome in patients with KS ; howeverp = 0.98). Alternatively, Schiff et al. (n = 5) who had previously been treated with exogenous androgens and to obtain the return of spermatogenesis after testosterone, patients may require a washout and recovery period of years. It has been reported that the median time to recover normal sperm production of 20 × 106 ml−1 sperm after discontinuation of testosterone therapy is between 3 and 6 months, with recovery in 100% of men at 24 months  or 14–16 years (37%) , though Other types of medications can be used to increase endogenous and, in particular, intratesticular testosterone production to stimulate spermatogenesis. The human chorionic gonadotropin (hCG) acts by stimulating partially functioning Leydig cells and increase testosterone production. Long-term use of hCG alone can induce spermatogenesis in a high percentage of patients but further improvement is generated with the addition of FSH. In studies performed in men with hypogonadotropic hypogonadism, trials of hCG alone (given for 14–20 months) resulted in the induction of spermatogenesis in 70%  and 50% Choice of TRT modality, administration, timing, hormone target levels, and monitoring are therefore important factors affecting FP outcomes. Further investigation into these therapies to improve spermatogenesis could be evaluated in a pediatric setting to maximize FP potential.Special considerations need to be made for the ethical and psychological aspects of exploring FP in childhood. In particular, ethical dilemmas surrounding FP in pediatric patients with KS ought to be discussed with patients and parents, and risk-benefit consideration must be evaluated prior to proceeding. During counseling, it is essential to address the rights of the children to an open future, perhaps justifying parental consent to explore FP measures in children with KS to provide the best opportunity to reproduce genetically in adulthood. Pediatric patients may need multiple consultations to understand the complex issues surrounding their future fertility, as was described by Rives et al.  who founIn the UK, storage of sperm is regulated by the Human Fertilisation and Embryology Authority (HFEA), which released the HFEA Act that outlines that patients under 18 years of age are able to consent to sperm storage if they are considered competent  . HoweverThese concerns, as well as the multi-system involvement of FP in KS, may be further addressed by utilizing a multi-disciplinary team (MDT) approach. Similar approaches have been utilized in pediatric FP in cancer patients and could be used as a foundation to build a similar model in patients with KS.There is growing evidence to support the benefit of carrying out FP in adolescent patients with KS, although controversies remain, and such practice is still in its infancy. A lack of large-scale studies is required to establish if such a practice should be recommended. Ethical dilemmas concerning FP in pediatric patients with KS remain, and discussions should be carried out with the patient and his parents in a multi-disciplinary setting to make shared decisions.CP, AC, AG, and WL wrote sections of the manuscript. CP formulated the first draft. All authors contributed to manuscript revision, read and approved the submitted version, and conception and design of the study.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


MappK = 0.116 ± 0.015 mM for dopamine and MappK = 0.245 ± 0.031 mM for L-epinephrine and linear dynamic ranges spanning up to 0.10 mM and 0.20 mM, respectively. Calculated limits of detection for both analytes were found to be within the sub-micromolar concentration range. The biosensor applicability to the assay of dopamine concentration in a pharmaceutical product was demonstrated .Engineering electrode surfaces through the electrodeposition of gold may provide a range of advantages in the context of biosensor development, such as greatly enhanced surface area, improved conductivity and versatile functionalization. In this work we report on the development of an electrochemical biosensor for the laccase-catalyzed assay of two catecholamines—dopamine and L-epinephrine. Variety of electrochemical techniques—cyclic voltammetry, differential pulse voltammetry, electrochemical impedance spectroscopy and constant potential amperometry have been used in its characterization. It has been demonstrated that the laccase electrode is capable of sensing dopamine using two distinct techniques—differential pulse voltammetry and constant potential amperometry, the latter being suitable for the assay of L-epinephrine as well. The biosensor response to both catecholamines, examined by constant potential chronoamperometry over the potential range from 0.2 to −0.1 V  showed the highest electrode sensitivity at 0 and −0.1 V. The dependencies of the current density on either catecholamine’s concentration was found to follow the Michaelis—Menten kinetics with apparent constants  Laccase is a complex copper-containing metalloproteinase found in trees, fungi and bacteria that exhibits enzymatic activity in the oxidation of aromatic or non-aromatic organics as well as some metallic ions in the presence of molecular oxygen . The enzLaccases immobilized on conductive materials have been found to be among the few enzymes capable of exchanging electrons with underlying electrode surface while electrocatalytically reducing oxygen to water via a four-electron mechanism . This poAnother way to improve the efficiency of the oxygen reduction reaction (ORR), even when laccase is immobilized in random orientation, is to add its substrates to mediate the electron transfer with the electrode surface . The resulting current rises proportionally to the mediator concentration, because the latter donates one electron to the T1 laccase active site; electricity flows further through internal pathways to the tri-nuclear cluster where the ORR takes place, this way facilitating the electron exchange with the electrode. Mediated oxygen reduction has been actively explored in an electroanalytical aspect as well, since diverse biosensors for phenols ,13,14,15Dopamine and epinephrine are structurally related catecholamines, playing the role of neurotransmitters (NT) in the human body, the abnormal levels of which in blood plasma or urine often indicate pathological conditions such as tumors or serious neurological disorders . DespiteElectrochemical biosensors offer a range of advantages over these techniques, such as selectivity, sensitivity and low limits of detection as demonstrated by Alvarez and Ferapontova  with theBoth catecholamines—dopamine and L-epinephrine—are organic substances of pharmaceutical importance since both compounds can be found on the drug market in the form of injection solutions. Therefore, the development of rapid and sensitive methods for their quantification is of immediate practical importance for pharma industry.Trametes pubescens immobilized on glassy carbon electrode modified with nanoporous gold. The main benefits from using nanogold-structured electrodes are the boosted sensitivity of the determination due to the enhanced surface area, and improved limits of detection resulting from the improved interfacial conductivity /K3[Fe(CN)6] as redox probe over the range of frequencies from 100 kHz to 1 Hz.Electrochemical impedance spectroscopy (EIS) was accomplished in a conventional 3-electrode setup, in 0.1 M KCl containing 5 mM KThe surface morphology of the modified electrode was examined with a scanning electron microscope JEOL JSM-6390 . No conductive coatings or other treatments were used on the sample prior to SEM observations.Before modification, glassy carbon electrodes were polished with 0.3 and 0.05 μm alumina slurry on a polishing cloth , water-rinsed and cleaned by ultrasonication in ultrapure water for 2–3 min.4, dissolved in 0.1 M HCl by cycling between 0 and −0.6 V  for 1 cycle at a scan rate of 0.1 V s−1.The working surface of the cleaned and polished glassy carbon electrodes was modified through direct electrodeposition of gold via electroreduction of tetrachloroaurate ion from an electrolyte containing 50 mM HAuCl2SO4 by cyclic voltammetry  over the potential range from −0.2 to 1.5 V  for at least 10 cycles, then thoroughly rinsed with ultrapure water.Prior to the enzyme immobilization gold-modified GC electrodes were cleaned electrochemically in 0.5 M HLaccase was immobilized as described previously , through−1 aqueous solutions. The duration of the sorption process was 2 h. After completing the chemisorption, the loosely bound alkanethiol was removed from the surface by rinsing with ultrapure water. Then, a 3 µL drop of laccase solution (16 mg mL−1) was cast on the electrode surface and 1 µL of glutaric aldehyde (45 mM aqueous solution) was mixed with it and allowed to react for at least 30 min at ambient temperature, then another 3 µL drop of laccase solution was cast to which a portion of 1 µL glutaraldehyde was added and allowed to react for another 30 min at ambient temperature. The prepared enzyme electrodes were stored in 0.05 M sodium citrate buffer, pH 4.0, in a refrigerator at 4 °C until use.The self-assembly of cystamine was carried out under static conditions by immersing the electrodes in its 10 mg mL−1 solution for injections and ampoule containing 1 mg mL−1 of L-epinephrine, or adrenaline. The analyses were performed under the optimized conditions and using pre-calibrated electrodes. The samples were spiked in the reaction medium (100 mL for dopamine assay and 20 mL for L-epinephrine assay) in volumes ensuring that the expected concentration will fall within the linear part of the calibration graph. Concentrations were determined on the basis of the calibration plots in molarities and converted to mg mL−1 so as to determine the recovery rates.The laccase biosensor was validated analyzing solutions with known concentrations: ampoule of dopamine hydrochloride, 40 mg mLAll the measurements were implemented at least in triplicate to ensure biosensor reproducibility.p). The calculated value for the Rp is much smaller than the one for the bare glassy carbon, implying that the electron transfer is facilitated on the gold-modified surface. The CPEs are often treated in the specialized literature as a typical characteristic of imperfect capacitors and suggests considerable surface roughness due to enhanced porosity or nanoparticles—modified electrode surfaces [Electrochemical impedance spectroscopy is known as a non-destructive alternating current technique capable of uncovering specific features of the studied systems. Complex plane plots of the impedance for an unmodified glassy carbon electrode and for the same electrode covered with electrodeposited gold are depicted in surfaces .2 from calculating the peak area by assuming an amount of 400 µC cm–2 electricity to desorb the chemisorbed oxygen [Scanning electron microscopy studies confirmed that, electrodeposited on glassy carbon, gold forms complex, rosette-like structures with dimensions between 50 and 130 nm. As can be seen from the SEM image C, the mod oxygen . The estLaccase immobilization on electrode surfaces is of key importance when aiming at a lasting operational stability of resulting bioelectrodes. To immobilize laccase on the Au electrodeposits, a positively charged at the working pH, self-assembled monolayer, SAM, of 2-aminoethane thiol was created (resulting from the chemisorption of cystamine on gold. This electrode preparation is further denoted in the text as cystamine-modified electrode). Laccase was further confined on the electrode surface by binding it to the thiol layer using the bi-functional agent, glutaric aldehyde.The electrochemical behavior of the immobilized enzyme was then studied by means of cyclic voltammetry (CV). Comparison of the CVs of the gold-modified electrode with immobilized laccase recorded in the absence and the presence of dioxygen  reveals 1 = 1.71) and protonated vicinal amino group bearing a positive charge [Differential pulse voltammogram (DPV) of the same laccase electrode is depicted in e charge , thus thThe appearance of a clearly expressed peak at the DPVs of laccase immobilized on the cystamine-functionalized gold layer, motivated us to further explore the voltammetric behavior of laccase in the presence of two structurally similar catecholamines—dopamine and L-epinephrine. The addition of dopamine aliquots to the buffer followed by the record of the resulting DPV A caused redE and oxE stands for the reduced/oxidized form of enzyme and redM and oxM the reduced and oxidized forms of catecholamine. The proposed mechanism was confirmed by comparative studies—in the absence of oxygen, no electroreduction of either catecholamine was observed.Further studies of the laccase electrode by cyclic voltammetry (CV), recorded in the presence of each catecholamine  showed aThe efficiency of the mediated electron transfer (MET process) was found to depend on substrate structure. Immobilized laccase is capable of reducing oxygen in the presence of dopamine at potentials more negative than 0.4 V rather efficiently . The potential region was selected so that its start point was the potential at which the DPV peak appears, and the end point is the potential at which the reductive current in the presence of L-epinephrine reaches a plateau. Over the whole studied potential region current changes stepwise upon addition of catecholamine stock solution . The tim−1 cm−2 (dopamine) and 0.12 µA L mol−1 cm−2 (L-epinephrine). This initial linear trend corresponds to the difusion-controlled region typical for the low substrate concentrations. At higher catecholamine concentrations, the dependence deviates from linearity  and the enzyme reaches saturation at dopamine concentrations exceeding 300 µM, whereas for L-epinephrine the saturation concentration was found to be a bit higher (above 350 µM). The apparent kinetic constants maxV and MappK were calculated from non-linear regression of the experimental data obtained with the two enzyme substrates, and are presented in MappK = 0.0918 mM as reported in [The electrochemical Michaelis–Menten plots  were draorted in ) and theThe apparent Michaelis’ constant for L-epinephrine determined under the same conditions was found to be more than twice as large, thus pointing to a weaker enzyme–substrate interaction. At concentrations exceeding 400 mM a slight decrease of the current density is observed—possibly indicating enzyme inhibition by L-epinephrine.−1) fall within this range [Operational parameters of the laccase biosensor when either dopamine or L-epinephrine were used as analytes are presented in is range .The discussed here, the operational parameters demonstrate a very good analytical performance of the developed biosensor in terms of electrode sensitivity and limits of detection. A brief comparison of the LOD values for some recently reported biosensors for dopamine (and L-epinephrine) assay is given in −1 for dopamine (40 mg mL−1) and 0.948 mg mL−1 for L-epinephrine (1 mg mL−1) support the biosensor applicability for the determination of these two analytes in pharmaceutical products.Validation of the laccase biosensor was implemented by analyzing pharmaceutical products with known concentrations . The detHaving in mind that gold nanoparticles are an excellent catalyst for ascorbate electrochemical oxidation at near zero potential, the presence of L-ascorbic acid in the assayed sample might potentially interfere with the electrode response to either catecholamine. Hence, the laccase electrode response to L-ascorbate was examined under the same experimental conditions at the operating potentials of 0 V and −0.1 V. It was found that at 0 V, the addition of L-ascorbate causes a reductive current to flow at an increased noise level . If presCalibration of the laccase electrodes towards either catecholamine was performed under isothermal conditions, at 20 ± 1 °C. The current density of the laccase biosensor in the presence of either catecholamine was found to be highly sensitive to temperature changes: an increase in the temperature by 6–7 deg. caused a drastic decrease in the electrode response. A plausible explanation is the fast auto-oxidation of catecholamines under ambient conditions and thus the substrate solutions were prepared immediately before measurement and kept in an ice bath during chronoamperometric measurements. Some contribution to the decreased reaction rate from the decreased oxygen solubility in buffers as temperature increases is also probable.Stability of fabricated biosensor was tested over a 26-hour period  using doA conventional glassy carbon electrode has been modified with porous gold through electrodeposition. The resulting modified electrode has an electrochemically accessible surface area that is ca. 12 times larger than the surface area of a smooth gold electrode with the same diameter.Trametes pubescens laccase immobilized on porous gold-modified electrode has been investigated by cyclic and differential pulse voltammetry in both absence and presence of oxygen. Voltammetric studies revealed laccase susceptibility to direct electron transfer and bioelectrocatalytic oxygen reduction. In the presence of catecholamines dopamine and L-epinephrine studies showed the capability of the immobilized laccase to perform mediated oxygen reduction with efficiency dependent on substrate structure.The electrochemical behavior of Amperometric measurements performed at a constant potential of −0.1 V  upon addition of either dopamine or L-epinephrine showed that the dependency of the current density on catecholamine concentrations follows the mechanism of Michaelis–Menten. The apparent kinetic constants have been found to depend on substrate structure with the values of The concentrations of dopamine hydrochloride and L-epinephrine in ampules with solution for injections has been determined with the developed biosensor. The analytical recovery of the determination has been found to be within 99% and 106% for dopamine and between 89% and 105% for L-epinephrine.Interference studies have shown that at an operating potential of 0 V the presence of L-ascorbate will affect the laccase electrode response to L-epinephrine by 1.1% and to dopamine by 0.8%, whereas at an operating potential of −0.1 V the interference from L-ascorbate will contribute a 2.2% increase in the response to L-epinephrine, and 1.4% to dopamine.Stability tests have shown that after 21 h storage in the refrigerator, the biosensor response to dopamine decays to 1/3 of the initial one.The discussed experimental findings demonstrate the plausibility of the chosen approach for laccase immobilization for the development of electrochemical biosensors for catecholamines capable of quantifying dopamine or L-epinephrine at close to physiological concentrations.


The properties of glass fiber composites were improved by strengthening the interlaminar regions using electrospun nanofibers mats. However, the chaotic nature of the electrospinning process at the collector restricts the controlled deposition and alignment of nanofibers and limits the use of electrospun nanofibers as secondary reinforcements. Hence, auxiliary vertical electrodes were used, which drastically reduced the diameter of the nanofibers from 450 nm to 150 nm and also improved the alignment of nanofibers. The aligned nanofibers were then used for doping the functionalized single-walled carbon nanotubes (f-SWCNTs) with nanofibers, which controlled the inherent issues associated with SWCNTs such as agglomeration, poor dispersion, and alignment. This process produced f-SWCNTs doped nanofiber mats. A series of tensile, three-point flexural, and Charpy impact tests showed that 30 vol% glass fiber composites reinforced with 0.5 wt% of randomly oriented nanofiber (RONFs) mats improved the properties of the hybrid composites compared to 0.1 wt%, 0.2 wt%, and 1 wt% RONFs mats reinforced glass fiber hybrid composites. The increase in properties for 0.5 wt% composites was attributed to the higher specific surface area and resistance to the relative slip within the interlaminar regions. The 0.5 wt% RONFs were then used to produce 0.5 wt% of continuous-aligned nanofiber (CANFs) mats, which showed improved mechanical properties compared to 0.5 wt% randomly oriented nanofiber (RONFs) mats reinforced hybrid composites. The CANFs mats with reduced diameter increased the tensile strength, flexural strength, and impact resistance by 4.71%, 17.19%, and 20.53%, respectively, as compared to the random nanofiber mats. The increase in properties could be attributed to the reduced diameter, controlled deposition, and alignment of the nanofibers. Further, the highest increase in mechanical properties was achieved by the addition of f-SWCNTs doped CANFs mats strengthened hybrid composites, and the increase was 30.34% for tensile strength, 30.18% for flexural strength, and 132.29% for impact resistance, respectively. This improvement in properties was made possible by orderly alignments of f-SWCNTs within the nanofibers. The SEM images further confirmed that auxiliary vertical electrodes (AVEs) reduced the diameter, improved the alignment and molecular orientation of the nanofibers, and thus helped to reinforce the f-SWCNTs within the nanomats, which improved the properties of the glass hybrid composites. Glass fiber composites have a major role in various engineering applications due to their design flexibility, ease of manufacturing, and low weight compared to conventional materials such as steel ,2. HowevResearch works ,8 show tSeveral studies focus on using various nanoparticles prepared as mats for reinforcing composites such as graphene-embedded paper for electromagnetic interference shielding (EMI) ,grapheneSeveral techniques are being used for producing polymeric nanofibers, and among them, electrospinning is considered simple and versatile. However, the polymeric jets formed during electrospinning have two distinct phases: near field, and far field . In the Hence, the focus of this study is to utilize the electrospinning setup with AVEs for producing CANFs mats and functionalized SWCNTs (f-SWCNTs) doped CANFs mats. The SWCNTs will be functionalized to improve the interfacial adhesion with nanofibers. Lastly, the effect of CANFs and f-SWCNTs doped CANFs mats on the mechanical properties of glass fiber composites will be evaluated using tensile, flexural, and impact tests to characterize the interlaminar strengthening mechanism.The polymer solution used in electrospinning was prepared by mixing Polyacrylonitrile powder  at the voltage range of 5 KV to 30 KV. Before analysis, the specimens were coated with a thin layer of carbon and a 15 nm layer of gold–palladium (Au/Pd) in a ratio of (60:40) in an EMITECH K950X evaporator and EMITECH K550X sputter coater, respectively. The specimens were then attached to 1 cm diameter stubs with DAG 580 colloidal graphite. The FTIR analysis was conducted on a Bruker Vector 22 FTIR spectrometer using 64 sample scans and 32 background scans. The Raman analysis was conducted using a Horiba Jobin-Yvon Lab RAM HR Raman spectrometer with a 514.5 nm line of an argon-ion laser as the excitation source.The SWCNTs were functionalized using the Friedel–Crafts alkylation (FCA) process. The FCA process was carried out by mixing SWCNTs (0.4 g) and PVA (4 g) with a 40 mL DMSO solution in a 250 mL flask using a magnetic stirrer. The flask was then immersed in an oil bath for gradual heating to 90 °C in the presence of nitrogen. As the heating promotes the chemical process, a catalyst of aluminum chloride  and functionalized SWCNTs (f-SWCNTs) and are shown in disorder  and defidisorder . In the  process .The electrospinning setup considered for this research consists of a DC voltage supply, NE-1600 syringe pump, and a grounded rotating drum collector (diameter 120 mm and width 210 mm), respectively. The electrospun nanomats were collected over the glass fiber mats attached to the rotating drum collector. The details of the complete setup with the spinning parameters were presented in our published manuscript  and the To analyze the effect of AVEs on the whipping effect, an electric field simulation software was used to simulate the electrostatic forces. The software developed the electrostatic forces as electric field lines (green lines) between the spinneret and the collector. The positive electrode (the spinneret) is red, while the blue color refers to the negative electrode. The simulation shows that without AVEs, the electric field lines are spread over the entire width of the collector. As the electric fields were not focused on the collector, the whipping effect deposited the nanofibers randomly over the whole width of the collector a. HoweveThe electrospinning setup was used to produce the required nanomats. At first, the randomly oriented PAN nanofiber (RONFs) mats were produced without AVEs for the weight fractions of 0.1%, 0.2%, 0.5%, and 1%, respectively. Based on the experimental results, the weight fraction of 0.5 wt% PAN nanofibers were selected to manufacture continuous-aligned nanofibers (CANFs) mats using AVEs. Finally, the 0.5 wt% CANFs mats were doped with 0.25 wt% f-SWCNTs to produce f-SWCNTs doped CANFs mats. For doping, only 0.25 wt% f-SWCNTs were selected to minimize the density effect following the results from Pilehrood et al. . Before The composites were fabricated using a vacuum-assisted resin transfer molding (VARTM) method. Following our previous work , 32 vol%The VARTM mold consists of a female square steel ring, and upper and lower male plates, and these were assembled with the alternative layers of woven E-glass fiber mats coated with the RONFs, CANFs, and f-SWCNTs doped CANFs mats. Then, the resin-hardener mixer was placed at the mold suction port and the vacuum pressure of 80 psi was maintained until the resin filled the mold. Finally, the resin was allowed to cure for 24 h at room temperature. The discussions of the results are divided into two sections. The first section will focus on analyzing the results from RONFs mats strengthened hybrid composite specimens, and the CANFs mats and f-SWCNTs doped CANFs mats strengthened hybrid composites will be discussed in the second section.The increase in the properties could be attributed to the reinforcing effect of RONFs mats due to their higher specific surface area, which could have increased the contact area within interlaminar regions. The infused RONFs mats provided more resistance to the relative slippage of the matrix over the glass fiber mats while transferring the applied load refer to a and thuHowever, a similar effect was not observed for further increase in RONFs mats from 0.5 wt% to 1 wt%. This may be associated with the nanofibers agglomeration and poor penetration of resin into the nanofibers network due to the random orientation with higher weight fractions, which could have acted as stress concentration sites. Based on the above results, 0.5 wt% of nanomats were selected to manufacture 0.5 wt% CANFs mats and 0.25 wt% f-SWCNTs doped with 0.5 wt% CANFs mats strengthened glass fiber hybrid composites specimens to understand the effect of alignment and f-SWCNTs doping within the interlaminar regions. The experimental results used 0.5 wt% of RONF mats strengthened hybrid composites as a reference, and the results are presented below.The tensile, flexural, and impact properties of CANFs mats and f-SWCNTs doped CANFs mats strengthened hybrid composites are shown in The phenomenal increase in mechanical properties was possible due to the reinforcement effect of continuous/aligned nanofibers along with uniformly dispersed f-SWCNTs doped CANFs mats within the interlaminar regions. The increase in mechanical properties can be attributed to the molecular orientations, reduction in nanofibers diameter, and the uniform dispersion of the aligned nanofibers network due to AVEs. Further, the reduction in nanofibers diameters (450 nm to 150 nm) provided an approximately threefold increase in nanofibers network within the interlaminar region.As discussed before, the bridging mechanism within the multiscale hybrid composites can be seen from the SEM images, as shown in The continuously aligned nanofiber mats with and without f-SWCNTs were produced using an electrospinning setup with AVEs, which were then used to reinforce the interlaminar regions of glass fiber composites. The effect of aligned nanomats on the mechanical properties of glass fiber composites was seen from the experimental results; as such, both CANFs and f-SWCNTs doped CANFs produced hybrid composites with improved properties by strengthening the interlaminar properties. The addition of AVEs with the electrospinning reduced the nanofiber diameter and improved their alignments within the interlaminar regions. The reduced diameter and alignment of nanofibers helped to improve the molecular orientation and further helped to distribute f-SWCNTs uniformly into the glass fiber hybrid composites. The functionalization of SWCNTs promoted better dispersion of SWCNTs and improved the bonding between f-SWCNTs and nanofibers. These factors promoted better load transfer within the interlaminar regions and thus improved the mechanical properties of the glass fiber composites. The research could serve in different applications, such as electronics, filtration, etc., where aligned nanofibers with reduced diameter and improved molecular orientation are the essential requirements. The application of SWCNTs could also be extended to other research areas, such as smart textiles, transistors, information storage devices, batteries, EMI shielding, etc.


This brief commentary aims to provide an overview of the available and relatively new precision management of reward deficiencies manifested as substance and behavioral disorders. Current and future advances, concepts, and the substantial evidential basis of this potential therapeutic and prophylactic treatment modality are presented. Precision Behavioral Management (PBM), conceptualized initially as Precision Addiction Management (PAM), certainly deserves consideration as an important modality for the treatment of impaired cognitive control in reward processing as manifested in people with neurobiologically expressed Reward Deficiency Syndrome (RDS). Blum and Noble  [The Precision Behavioral Management (PBM) platform is related to addiction medicine, with the first USA and foreign patents related to the accurate Genetic Addiction Risk Severity  ,101,102.A, 1990) ,125,126.homo sapiens with a usual happiness setpoint  [The cascading interaction of neurotransmitters and second messengers results in the correct release of dopamine within the NAc and across many brain regions. These regions are involved in motivation, cognition (memory), pleasure, stress reduction, drug reinstatement, decision making, recall, wellbeing, and cravings. The result is to provide ostasis) , reflectostasis) .In the neurophysiologic reward system, repeated frequent acute dopamine stimulation becomes chronic stimulation and leads to a dysfunctional hypodopaminergic state, rendering the reward system less responsive to natural reinforcers, a symptom of RDS ,128. TheAs alluded to above, reward deficiencies may also occur in the absence of dopaminergic stimulation by exogenous factors due to specific polymorphic alleles that alter the function of genes in the reward cascade. One example is the A1 allele of the D2 dopamine receptor gene that causes a reduced number of dopamine receptors in the mesolimbic NAc. An essential feature of RDS is the lack of integration between cognition, perception, and emotions occurring due to (1) substantial dopaminergic surges in reward, motivation, and learning centers leading to neuroplasticity in the striato-thalamic-frontal cortical loop, with ensuing top-down dissociation from the subcortical activity; (2) hypo-functionality of the excitatory glutamatergic afferents from the amygdala–hippocampus complex failing to produce bottom-up restraint of the striato-thalamic-frontal cortical loop ,133,134.The above aberrations may be a target of neuromodulation with therapeutic interventions and prophylaxis of addictive and related disorders. PBM combines the GARS test that identifies RDS risk polymorphisms and is used to ascertain the neuropathways involved in the tested individual’s hypodopaminergic risk neurotransmission finite pathways. These pathways are used in an algorithm to select a neuronutrient formulation of KB220 for that individual to induce the desired dopamine homeostasis by balancing genetic and epigenetic (neuro-molecular) brain reward activity .The GARS test uses saliva samples and polymerase chain reaction (PCR) to identify dysfunctional polymorphisms of reward genes . Genes eHypo-dopaminergia refers to a reduced dopamine function in the brain reward circuitry. As stated in the paper and indicated by the interrelatedness of at least seven main neurotransmitter pathways involving synthesis, vesicular pre-neuronal storage, mitochondrial catabolism, synaptic catabolism, neuronal clearance via transporters, receptor affinity, and number, carrying any one of the eleven polymorphic alleles could explain low dopamine function see . One exap < 0.04 predicted with seven or more alleles, and drug risk severity p < 0.05 predicted with four or more alleles. Understanding the genetic antecedents to Alcohol Use Disorder (AUD) may help explain potential neuroplasticity in those dependent on alcohol or other substances of abuse [The GARS test examines the sum of many related polymorphisms instead of one gene alone to predict genetic risk for RDS behaviors. It is possible to find a significant association with the degree of risk for all addictive behaviors (RDS). In conjunction with the Institute of Behavioral Genetics at Colorado University, unpublished research compared the GARS test with the Addiction Severity Index (ASI) in 273 subjects from substance treatment centers to determine drug and alcohol risk severity. Alcohol risk severity of abuse .The PBM system uses the patented, commercially available GARS test with a nutraceutical Precision Neuronutrient-Research ID code: KB220 ,145,146.The GARS test identifies RDS risks and determines the neuropathways involved in hypodopaminergic risk to identify the neuronutrient formulation of KB220 via algorithm and create balanced genetic and epigenetic (neuro-molecular) brain function, the desired induction of dopamine homeostasis.This Special Issue is not only about risky behaviors (addiction) but also cognitive control of reward processing; here, we provide a few examples, emphasizing the role of dopamine in addiction and cognition. The neuronal release of dopamine in the limbic system relies on many neurotransmitters, peptides, and second messengers to impact the dopamine released at the NAc, which is critical to feeling good .There are many examples of cognitive control of various aspects of drug and non-drug addictive behaviors. For example, the ability to resist the urge to eat is in part a function of the homeostatic functioning of neuronal circuits involved in top-down control to oppose the conditioned responses that predict reward from eating the food and the desire to eat the food. Moreover, imaging studies by Volkow’s group of this non-drug behavior have revealed that obese probands have impairments in dopaminergic pathways that regulate neuronal systems linked with reward sensitivity, conditioning, and control . There iSubstance use disorders (SUDs), while effecting billions worldwide, are indeed preventable. Thousands of articles attest to the well-known effects of substance misuse that cause many physiological, molecular, and cellular changes in specific brain regions. Moreover, these neuroplastic changes have a role in seeking behaviors seen in substance and non-substance addictions ,149.Notably, many studies have focused on the dopamine neurons of the ventral tegmental area (VTA) and the regions where these neurons terminate: the striatum, the prefrontal cortex , and theAnother example is the role of cognitive control and reward processing in Internet Gaming Disorder (IGD). There are indeed common neurochemical mechanisms that have been observed with both IGD and SUD . SpecifiThere is also evidence from experiments that used lentivirus tools for over-expression or silencing of the dopamine transporter (DAT) gene in animals. Behavioral profiles that evaluated motivation and self-control compared to controls revealed significant differences. Specifically, DAT over-expressing rats showed increased impulsivity. The authors concluded that an attenuated dopaminergic tone following altered accumbal DAT function may subserve a sensation-seeker phenotype with vulnerability to lack of impulse control ,162,163.No matter what therapeutic strategy the clinical team chooses, a beneficial practice for treatment and recovery is genetic addiction risk testing and personalized induction of dopamine homeostasis based on genetic test results.Indeed, the use of any treatment that reduces stress and enhances resting-state functional connectivity along the brain reward circuitry seems prudent.From pre-authorization of very short-term use of opioids to reduce harm to cognitive behavioral therapy, trauma therapy, brain spotting, stress reduction, rsTMS to deep brain stimulation, and 12-stepping, the foundational induction, via epigenetics, of gentle up-regulation of dopaminergic function will be an evidenced basis for the induction of treatment (without complications or side effects) to keep people from any form of dopaminergic dysfunction.Genetic addiction risk testing for patients attending a pain clinic provides information about the likelihood of a predisposition for Opioid Use Disorder (OUD) and enables the utilization of less addicting analgesia at the onset of treatment.Genetic testing should be the standard of care for all patients attending substance and non-substance (process addictions) dependency programs .Genetic addiction risk testing for early risk identification in children, especially if they have addiction issues in the family , combined with precision pro-dopamine regulation prophylaxis, may attenuate or prevent addiction risk.Coupling of KB220 precision variants with naltrexone to improve compliance .Combat cannabis-induced anhedonia in adolescents and adults with RDS using “Precision Behavioral Management” (PBM) to provide precision KB220 formulations ,146.In summary ,165,166:Billions worldwide are affected by SUDs and RDS, which are preventable with PBM, genetic testing for addiction risk, and pro-dopamine precision KB220 formulations selected to treat neurotransmitter deficits. The neurotransmitter deficits are caused by the tested person’s dysfunctional alleles, identified by their genetic test results. This required induction of “dopamine homeostasis” is recommended for frontline tertiary addiction treatment and relapse prevention.While more in-depth and extensive double-blinded studies combining the GARS test with KB220 precision formulations are encouraged, we believe that this body of evidence presented to support PBM, an important therapeutic and prophylactic modality for the treatment of impaired cognitive control of reward processing as manifested in patients expressing RDS, deserves careful consideration.


Voxel-based morphometry was used to investigate associations between neurochemical measures and gray matter volumes. Whole striatum D2/3R availability was positively associated with prefrontal cortex gray matter volume (pFWE corrected = 0.048). This relationship was mainly driven by associative receptor availability (pFWE corrected = 0.023). In addition, an interaction effect was observed between sensorimotor striatum D2/3R availability and anterior cingulate Glx, such that in individuals with greater anterior cingulate Glx concentrations, D2/3R availability was negatively associated with right frontal cortex gray matter volumes, while a positive D2/3R-gray matter association was observed in individuals with lower anterior cingulate Glx levels (pFWE corrected = 0.047). These results are consistent with the hypothesis that the prefrontal cortex is involved in regulation of striatal dopamine function. Furthermore, the observed associations raise the possibility that this regulation may be modulated by anterior cingulate glutamate concentrations.Prefrontal cortex has been shown to regulate striatal dopaminergic function via glutamatergic mechanisms in preclinical studies. Concurrent disruption of these systems is also often seen in neuropsychiatric disease. The simultaneous measurement of striatal dopamine signaling, cortical gray matter, and glutamate levels is therefore of major interest, but has not been previously reported. In the current study, twenty-eight healthy subjects underwent 2 simultaneous [ An improved understanding of the relationship between cortical structure and neurochemistry has the potential to advance our pathophysiological understanding of diseases in which simultaneous disruptions exist .The striatum exhibits a high degree of connectivity to both the cortex and other subcortical structures , 3, and 2 receptor availability in healthy subjects -( + )-PHNO is able to index striatal D2/3 receptor availability, and, in conjunction with amphetamine administration, can measure dopamine release -( + )-PHNO PET-MRI in a sample of healthy participants. Specifically, we hypothesized to find a direct relationship between PFC gray matter volumes and striatal dopamine receptor availability, an inverse relationship between PFC gray matter volumes and amphetamine-induced dopamine release, and these relationships to be modulated by ACC Glx concentrations.In the present study, we set out to investigate relationships between prefrontal cortex (PFC) gray matter volumes, anterior cingulate cortex (ACC) Glx levels, striatal dopamine receptors, and striatal dopamine release using a simultaneous [1H]-MRS measures have not been previously reported.The study was approved by the local National Health Service (NHS) Research Ethics Committee (12/LO/1955) and the Administration of Radioactive Substances Advisory Committee. PET data were previously reported in McCutcheon et al. , but theTwenty-eight participants were enrolled through online advertising. Inclusion criteria were age 18 or above and ability to give informed consent. Exclusion criteria included contraindications to MRI or PET scanning  or amphetamine administration , current or previous significant medical co-morbidity, past traumatic brain injury with loss of consciousness, background of neurological condition or psychiatric disorder (including substance use disorder) as determined by medical review and the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders IV (DSM-IV), and history of psychiatric disorder in first-degree relatives. Information concerning tobacco and cannabis use was collected. Subjects smoking at least one cigarette daily were considered smokers, five subjects had consumed cannabis at least once and only one subject regularly used on a weekly basis. All subjects provided informed written consent.Participants were asked to attend two imaging sessions separated by a minimum of 3 days, during which simultaneous PET-MRI scans were acquired. One session was performed under placebo condition (101 mg of lactose/sucrose tablets), the other after oral dexamphetamine administration (0.5 mg/kg). Scanning was initiated 3 h after placebo or amphetamine administration , 25. Pla11C]-( + )-PHNO over 30 s , PHNO is a dopaminergic agonist binding D3 receptors and high affinity D2 receptors [11C]-( + )-PHNO PET has an excellent reproducibility with intraclass correlation >0.9 for the striatum/pallidum -MRS data, a 20 × 20 x 20 mm voxel was placed midline on the anterior cingulate (ACC) region  = 400 ms, TE = 3.2 ms, repetition time (TR) = 8.5 ms, flip angle = 12°, 1 mm isotropic voxels, matrix = 256 × 256, and number of slices = 188. To acquire [The Molecular Imaging And Kinetic Analysis Toolbox (MIAKAT) version 4.3.13 in MATLAB 8.2  was used for PET data analysis . The motND) of [11C]-( + )-PHNO was estimated at each voxel -( + )-PHNO BPND to estimate D2/3 receptor availability and dopamine release is well validated, our cross-over design could lead to carry-over effects for subjects who received amphetamine first. A 3-day washout period should, however, allow for good elimination of a product with a 10 h half-life [While the use of [alf-life . Despitealf-life . Despite1H]-MRS is unable to distinguish between intra- and extra-cellular glutamate limiting the precision of possible inferences [It is not possible with current neuroimaging measures to identify which precise gray matter component underlies the observed relationship between striatal dopamine and gray matter volume. In addition, although participants were screened regarding cannabis use, past substance use may still have been unreported and may have the potential to contribute to the observed findings. While smaller gray matter volumes are often equated to fewer projection neurons, intracortical neuropil, glial cells, interneurons could also contribute to the observed findings. Similarly, [ferences . AlthougIn the current study, we demonstrated that the availability of dopamine receptors in the bilateral whole striatum and in the associative subregion positively correlate with right prefrontal gray matter volume, and that there was a negative interactive effect of ACC Glx on a right frontal gray matter volume. These results extend previous findings in schizophrenia and healthy participants to demonstrate interaction across all three systems, with relevance for our understanding of neuropsychiatric disease in which abnormalities are often found to occur simultaneously.Supplementary informationSupplementary Table 1Supplementary Table 2Supplementary Table 3Supplementary Table 4Supplementary Figure 1Supplementary Figure 2Supplementary Figure 3Supplementary Figure 4Supplementary Figure 5Supplementary Figure 6


Since COVID-19 was declared a pandemic by the World Health Organization on March 11, 2020, the disease has had an unprecedented impact worldwide. Social media such as Reddit can serve as a resource for enhancing situational awareness, particularly regarding monitoring public attitudes and behavior during the crisis. Insights gained can then be utilized to better understand public attitudes and behaviors during the COVID-19 crisis, and to support communication and health-promotion messaging.The aim of this study was to compare public attitudes toward the 2020-2021 COVID-19 pandemic across four predominantly English-speaking countries  using data derived from the social media platform Reddit.We utilized a topic modeling natural language processing method . Topic modeling is a popular unsupervised learning technique that can be used to automatically infer topics  from a large corpus of text. We derived our data from six country-specific, COVID-19–related subreddits . We used topic modeling methods to investigate and compare topics of concern for each country.Our consolidated Reddit data set consisted of 84,229 initiating posts and 1,094,853 associated comments collected between February and November 2020 for the United States, the United Kingdom, Canada, and Australia. The volume of posting in COVID-19–related subreddits declined consistently across all four countries during the study period (February 2020 to November 2020). During lockdown events, the volume of posts peaked. The UK and Australian subreddits contained much more evidence-based policy discussion than the US or Canadian subreddits.This study provides evidence to support the contention that there are key differences between salient topics discussed across the four countries on the Reddit platform. Further, our approach indicates that Reddit data have the potential to provide insights not readily apparent in survey-based approaches. In December 2019, several cases of respiratory disease were reported in Wuhan City, China . The numTo suppress the transmission of COVID-19, governments have enforced several waves of border shutdowns, travel restrictions, quarantine, and other nonpharmaceutical interventions such as mask mandates, limiting public activities, and restricting travel -7, sparkTraditional surveillance systems, including those utilized by the US Centers for Disease Control and Prevention and the European Influenza Surveillance Scheme, rely on both virologic and clinical data, and publish data once per week, typically with a 1-2–week reporting lag . Survey Social media such as Reddit, Twitter, Facebook, Weibo, and others provide a readily available source of abundant, organic, publicly accessible first-person narratives -25, whicDuring lockdown events, social media platforms have—through their individual users—provided informational support and online access to services for pregnant women to obtain prenatal care services, such as consulting and scheduling necessary appointments . SimilarTopic modeling, a popular statistical unsupervised machine-learning technique, has been widely used for discovering the underlying themes that occur in collections of health-related texts . Becauser/CoronavirusUK, r/CanadaCoronavirus), which enables a more targeted analysis of users from different countries [Reddit is one of the most popular social media platforms with over 430 million active users and 1.2 million subreddits  as of May 2020, with over 70% of its user base coming from English-speaking countries ,52. SomeIn this work, we employed Reddit data from six geographically specific COVID-19–related subreddits representing four English-speaking countries, the United States, the United Kingdom, Canada, and Australia, to investigate (1) whether there were key differences between salient topics discussed across the four countries and (2) whether Reddit data have the potential to provide insights not readily apparent in survey-based approaches. In general, LDA topic modeling was applied to each country-specific Reddit data set. We trained multiple topic models for each country consisting of a different number of topics and manually inspected each model to find the optimal model for each country . We further compared the summarized topics for each country based on each country’s model, and mapped them to four common topic categories . Finally, longitudinal topic trends were examined to identify trends in the common topic categories, which were then mapped to the COVID-19 events for each country.r/CoronavirusUK, r/coronavirusus, r/CoronavirusCanada, r/CanadaCoronavirus, r/CoronavirusAustralia, r/CoronavirusDownunder), as shown in As Reddit data do not generally include geolocation information, we collected data from the six most popular subreddits  related to the United States, the United Kingdom, Canada, and Australia , which means they are the most active and popular geographically specific COVID-19–related subreddits available. As users typically present their own experiences in the initiating posts [The consolidated Reddit data set consisted of 84,229 initiating posts and 1,094,853 associated comments collected between February and November 2020 derived from the six subreddits shown in ng posts , with sur/CoronavirusAustralia and r/CoronavirusDownunder. We then automatically identified URLs and email addresses, which were removed from the texts of submissions to simplify the subsequent topic modeling process. To remove the stop words , we first used the Natural Language Toolkit (NLTK 3.3 for Python 2.7) [Let’s go! was tokenized into the list “let,” “’s,” “go,” “!”) and lemmatized  using the Python SpaCy 2.2.1 package [cough, coughing) into a canonical form .To build the corpus for each country, we organized the submissions from the six subreddits shown in hon 2.7)  to initihon 2.7) . Subsequ package  to conveWe used the topic modeling technique to compare the broad themes emerging from the United States, the United Kingdom, Canada, and Australia. The general procedure is described in We trained multiple topic models  for each of the four countries using the LDA implementation in the Gensim 3.8.3  toolkit.For each topic model, the most characteristic keywords associated with each of the thematic topics were manually examined, focusing specifically on the posts that were particularly representative according to the contribution probability of those topics to determine which model best characterized the data set. In the process of manual identification of topics, we noticed that the models for the four countries had different optimal numbers of coherent, nonoverlapping topics. Further, some models contained topics idiosyncratic to that country . For example, the “mental health” topic in the UK topic model did not appear in the US topic model. To compare and contrast the common themes among the four countries, we consolidated these various topics into four common topic categories. Topics and their mappings to the common topic categories are listed in The prevalence of common topics for the United States, the United Kingdom, Canada, and Australia was studied by first finding the “document-topic” for each post. The document-topic refers to a topic that is the major constituent (according to the contribution probability) of a given document , which cUsing the document-topic threshold for each country, we identified the proportion of each topic by first calculating the number of posts whose topic probability was above the threshold, and then dividing this number by the total number of posts to obtain the topic proportion. The proportion of the common topic categories was determined by summing the proportion of the topics that belonged to each common topics category.With the document-topic threshold for each country, we also calculated the number of submissions on a specific common topic category for each week, before counting the weekly volume of submissions on each common topic category, to plot the common topic trend for each country from February to November in 2020. We also mapped the COVID-19 event timeline from the WHO  and ThinWe restricted our analysis to publicly available discussion content and the University of Utah’s Institutional Review Board exempted the study procedure and data from ethical review (IRB_00076188) under Exemption 2 as defined in the United States' Code of Federal Regulations (CFR), 45 CFR 46.101(b).Our COVID-19 Reddit data set comprises 10 months of discussions (February 2020 to November 2020), which covers the main early COVID-19 events, including the initial outbreak and subsequent lockdowns in the United States, the United Kingdom, Canada, and Australia. During this time, 103,180 unique users posted some 84,229 submissions and their associated 1,094,853 comments. To further study the behavior of posting on Reddit, we summarized the weekly post volume and user volume for each country, as shown in After manually examining the topic models  for the United States, the United Kingdom, Canada, and Australia, we qualitatively identified the most coherent model, as well as the threshold of the document-topics for each country-related data set, as shown in For each topic in each model, we mapped that topic to four common topics  very similar posts  may be assigned to different topics and (2) very simple posts  may correlate to many topics. Similar problems were discovered by Xu et al  when anaAnother issue in this work is related to the completeness of the Reddit data we collected via the pushshift.io API . Althougr/CanadaCoronavirus are mostly Canadians). Thus, the subreddits not only reflect people’s opinions but also the culture differences in the four countries. For example, people in the United Kingdom concentrate on discussing politics or COVID-19–related breaking news. Thus, the leading topic, politics-related policies, in r/CoronavirusUK does not fully reflect people’s concerns related to COVID-19, as it may simply reflect people’s discussion habit in the United Kingdom. Therefore, the differences in topics may not fully reflect people’s opinion toward COVID-19 in the United States, the United Kingdom, Canada, and Australia.A further limitation is related to the differences in culture associated with different subreddits. As Reddit data do not in general include geolocation information, we collected data from the six most popular COVID-19 subreddits related to the United States, the United Kingdom, Canada, and Australia. We examined the posts and noticed that most users are local people  of RIn this work, we used Reddit data to examine variations in people’s concerns during the COVID-19 crisis in the United States, the United Kingdom, Canada, and Australia. We found that people posted more on Reddit during lockdown events, and people’s concerns differ among the four countries. Further, this work provides evidence to support the contention that there are key differences between salient topics discussed across the four countries on the Reddit platform. Further, our approach indicates that Reddit data have the potential to provide insights not readily apparent in survey-based approaches.


However, topical drug application for hypertrophic scars are unsatisfactory. The purpose of this study was to explore the permeability of silk nanofiber hydrogels (SNFs) loaded with rhodamine 6G (R6G) and rhodamine 110 (R110) mediated by COIn vivo rabbit ear hypertrophic scars were treated with CO2 fractional laser irradiation and then R6G/R110-laden SNFs were applied to the scars to evaluate their synergetic effect on drug penetration efficiency. Their permeability was quantified by fluorescence intensity and measured by confocal laser scanning microscopy on days 1, 3, 5 and 7. More specifically, the thermal coagulation zone (CZ) and its surrounding area (peri-CZ) caused by the thermal coagulation of the laser were discussed separately.In this work, R6G and R110 were chosen as hydrophilic and hydrophobic model molecules. They were loaded inside SNFs. Our data indicated that the SNFs promoted the penetration of R6G but not that of R110 in the peri-CZ on day 1 when combined with laser irradiation. Interestingly, both R6G and R110 were abundant in the CZ and remained stable on days 1, 3 and 5. Moreover, rapid re-epithelialization hindered the long-term permeability of both drugs.2 fractional laser irradiation with SNF drug delivery could improve the efficiency of hydrophilic drug delivery within 24 h before total re-epithelialization.Combining CO Highlights2 fractional laser irradiation due to their good loading capacity for both hydrophilic and hydrophobic drugs.Silk nanofiber hydrogels provide a suitable platform for revealing the permeability of different drugs into skin treated with CO2 fractional laser irradiation was shown to enhance the permeability of hydrophilic and hydrophobic drugs in rabbit ear hypertrophic scars.CO2 fractional laser irradiation could improve the skin penetration of hydrophilic drugs within 24 h after laser treatment .Silk nanofiber hydrogels combined with COAbnormal wound healing usually results in hypertrophic scars (HSs) or even keloids ,2. HSs c2 fractional laser has been confirmed to improve the elasticity, thickness, appearance and symptoms of mature hypertrophic burn scars [in vitro experiments without long-term observation. Therefore, dynamic drug distribution over time has not been observed [2 fractional laser-assisted drug delivery [2 fractional laser-assisted drug delivery for the treatment of scars deserves more research attention with regard to enhancing the penetration of drugs.Based on dot-matrix photothermolysis theory, ablative fractional lasers (AFLs) have been proven to be able to increase the dermal absorption of topically administered drugs ,14 by dirn scars . Howeverobserved . Additioobserved , short ddelivery ,26. As s2 fractional lasers to achieve better transdermal delivery.In recent years, multiple nanocarriers with transdermal capacities have been developed and applied in AFL-assisted drug delivery, improving the therapeutic effect of drugs on HSs . Silk na2 fractional laser-mediated drug-laden SNFs could penetrate HSs more effectively. Water-soluble rhodamine 6G (R6G) and water-insoluble rhodamine 110 (R110) as hydrophilic and hydrophobic drug models were loaded on SNFs. When treatment with a CO2 fractional laser was applied in a rabbit HS model, the drug-laden SNFs were applied to the HSs to evaluate transdermal penetration in vivo. Additionally, the week-long in vivo experiment provided a link between the healing of laser channels and drug penetration for future studies.In this study, we aimed to reveal whether COThe original silk fibroin solution (6 wt%) was prepared through previously reported procedures . It was Hydrophobic and hydrophilic drugs were loaded on SNFs via various processes. For hydrophilic R6G (Acros Organics), the drug was added to the SNF solution directly and then loaded on the nanofibers after stirring for 4 h at room temperature. Hydrophobic drugs were loaded on the SNF through a blending–centrifuging process. R110  was dissolved in ethanol and then blended with SNF aqueous solution. The blend solution was stirred for 24 h at room temperature to induce the transfer of R110 from ethanol to the SNF. Then, the blend solution was centrifuged at 10000 rpm for 20 min to separate the R110-laden SNF from ethanol. The specific steps were as follows. R6G (3 mg) was dissolved in 100 μl of deionized water and the solution was added to 10 ml of 2 wt% SNF, immediately followed by 4 h of stirring using magnetic stir bars at room temperature. R110 (18 mg) was dissolved in 1 ml of 100% ethanol and mixed completely. The solution was slowly added to 20 ml of 2 wt% SNF and swirled for 24 h at room temperature. Then, the mixture was transferred to a centrifuge tube and subsequently centrifuged at 10000 rcf for 20 min to obtain molecule-loaded nanofibers. The supernatant was collected after washing with deionized water three times for the quantitative analysis of the unloaded R110 content using a UV–vis spectrophotometer . Through optimizing the conditions, the R6G and R110 loading capacities were 0.03 and 0.18%, respectively. R110@SNF (0.18%) was diluted with 2 wt% SNF to eventually obtain 0.03% R110@SNF. For the control, we also prepared a 0.03% R6G aqueous solution and a 0.03% R110 ethanol solution. The whole process was shielded from light.−1 at 25°C.The sample morphology was examined by atomic force microscopy . Samples were diluted to 1:1000 and dispersed in ultrasound for 10 min. Then, a 2 μl solution was spin-coated onto a clean mica surface and the surface was blown with nitrogen. A confocal Raman spectrometer  was used to evaluate the loading of R6G and R110 in the SNFs. The rheological properties of all hydrogels were measured with a rheometer . Frequency sweeps were collected continuously over a wide frequency range from 0.1 to 100 rad s3), and subcutaneous fat was removed and subsequently soaked in cell culture medium (Gibco) containing 2% penicillin–streptomycin (Gibco) and 10% foetal bovine serum (Gibco) for 30 min. Next, the small tissues were minced and digested in cell culture medium containing 0.2% collagenase  for 2–4 h in a 37°C incubator. Afterwards, the cell suspension was filtered through a 70 μm filter and centrifuged at 1200 rpm for 5 min. Finally, the cell pellet was resuspended in cell culture medium (Gibco) supplemented with 10% foetal bovine serum (Gibco) and 1% penicillin–streptomycin (Gibco), and the cells were cultured at 37°C in 5% CO2 with the medium changed every 3 days. For experiments, the cell passage was ≤5.Human keloid tissues were obtained from 18- to 45-year-old Chinese plastic surgery patients with informed consent at Shanghai Ninth People’s Hospital. To collect human dermal fibroblasts (HDFs), foreskins were obtained from Chinese circumcision patients with informed consent at Shanghai Ninth People’s Hospital. This study was performed according to the ethical guidelines of the 1975 Declaration of Helsinki and approved by Shanghai Ninth People’s Hospital. The human tissues were rinsed in Dulbecco’s phosphate buffered saline, the epidermis tissue was cut into small pieces . Human keloid fibroblasts (HKF) and HDF cells were seeded in 96-well plates with 2000 cells per well. After 24 h of incubation, they were treated with 0.5, 1, 2, 4 or 8 mg/ml SNF dissolved in cell culture medium for 1, 3, 5 or 7 days. At the corresponding time, cell culture medium containing 10% CCK-8 solution was added to each well and incubated for 1 h at 37°C. Finally, the absorbances were obtained at 450 nm using a microplate reader .Two-week-old male New Zealand white rabbits weighing 2 kg  were kept individually. Xylazine Hydrochloride was combined with 1.5 mg ketamine per 100 g weight of rabbits and administered intramuscularly. All of the wounds were located around the center point of the rabbit ears and were distributed equally. Each circular wound was removed with a diameter of 1 cm of full-thickness skin and perichondrium while the cartilage remained intact. The rabbit ear wounds healed and were repaired entirely, with a noticeable diameter of ~0.9 cm of HSs after 28 days. The scars appeared as bright red bumps on the skin, with obvious hyperplasia and characteristics of being thick and hard.2 fractional laser treatment . The laser was operated in DeepFX mode under the parameters of 25 MJ energy intensity, 20% coverage, 300 Hz emission frequency and on the 10th spot size without overlap. The rabbits were well anesthetized before laser irradiation via intramuscular injection anesthesia. All groups were subsequently given the corresponding drug treatments (50 μL drug/HS). After 1, 3, 5 and 7 days of treatment, rabbits were sacrificed by intravenous injection of an overdose of sodium pentobarbital and HS tissues were excised. These HS tissues were then cut into frozen sections with a 7 μm thickness. Finally, the frozen sections were viewed under a Zeiss LSM 880 confocal microscope . The fluorescence intensity was measured by ZEN 2.6 software.Mature rabbits with hypertrophic ear scars were randomly divided into eight groups: the control groups received topical treatment with R6G, R6G@SNF, R110 or R110@SNF, and the laser irradiation groups additionally received topical COP values <0.05 were considered statistically significant.All error bars represent standard deviations. Data analyses were conducted using GraphPad Prism 9.0 software. When comparing more than two samples, either one- or two-way analysis of variance was used, with Tukey’s multiple comparisons test. −1, the Raman spectrum of R6G@SNF exhibited the typical peaks of R6G at ~1367 and 1515 cm−1, while that of R110@SNF showed the typical peaks of R110 at ~1368 and 1506 cm−1 . This indicated that R6G@SNF could significantly enhance the penetration of R6G with the beneficial application of a CO2 fractional laser within 24 h. However, the effect of SNF was less effective in the following days. For the groups without laser treatment, the dermis was almost completely impermeable to R6G@SNF and R6G, and no significant difference was found between the R6G@SNF and R6G groups (The successful rabbit ear HS model is shown in G groups . Additiop < 0.001). However, the results for these two groups on day 3 were reversed (p < 0.01). This indicated that the effects of the R110@SNF treatment may be sustained longer on intact skin. For the laser treatment groups, R110 was mainly concentrated in the CZ , which was the zone ablated by the laser showing an ‘empty’ zone of the laser channel, and the other was the CZ, a rim of thermally coagulated tissue of variable thickness surrounding the individual microchannels, which reflected residual thermal damage [in vivo in a rabbit ear HS model for up to 7 days, with dynamic observation of drug delivery through confocal laser scanning microscopy. In particular, we provided a comprehensive comparison of the efficacies of CO2 fractional laser and SNF treatment regarding drug penetration in HSs. In addition, we studied the permeation of hydrophobic and hydrophilic drugs separately to provide a more comprehensive analysis of the permeation of both types of drugs in the laser channels and full-thickness skin. Collectively, our study concluded that SNF promoted the permeability of R6G in the laser channels within 1 day, but the same was not concluded for R110.This study characterized the distribution of SNFs loaded with R6G and R110 as hydrophilic and hydrophobic model drugs based on COl damage . Many ea2 fractional laser treatment had the best penetration in the peri-CZ at day 1 compared to the other three groups. This result indicated that CO2 fractional laser irradiation was a key factor for drug penetration. CO2 fractional laser irradiation not only removes the epidermal barrier directly but also enhances the total area of absorption [In our experiments, R6G@SNF with COsorption ,36. Thessorption . The presorption . A recensorption . Some drsorption . Likewissorption ,39,40, a2 fractional laser irradiation, to some extent, was beneficial for the direct entrance of R110@SNF and R110 into the skin. Moreover, due to hydrophobic interactions, this penetration was difficult to enhance. Similarly, Olesen et al. found that the permeability of the highly lipophilic drug vismodegib is only modestly enhanced by CO2 fractional laser irradiation [Meanwhile, for hydrophobic molecules such as R110, our study found that although R110@SNF and R110 solution could fill the channels, their penetration into the peri-CZ tissue was difficult. This illustrated that COadiation . Additioadiation . Improveadiation . Althoug2 fractional laser irradiation on increasing drug penetration began to fade after 24 h [et al. argued that a CZ of an appropriate thickness could serve as a reservoir for a small hydrophilic drug, promoting drug diffusion into the surrounding skin [in vitro. However, in this in vivo study, we found that for a long time, drugs could not permeate through the CZ. We speculated that rapid re-epithelialization of the CZ may be the key to unsustainable drug release. Another potential reason might be that the function of vascular/lymphatic systems and metabolism is to contribute to drug elimination [In previous investigations, the effect of COter 24 h , which wter 24 h . Our fining skin . Researcing skin . In our mination .2 fractional laser irradiation and topical 5-FU or verapamil hydrochloride treatment resulted in a significant scar treatment effect compared to the control group [Recently, a randomized clinical trial found that no significant long-term difference in scar flattening was observed between a laser + corticosteroid group and the control group . Howeverol group . We spec2 fractional laser-assisted drug delivery is more suitable for the delivery of hydrophilic drugs. In addition, SNF could serve as a penetration enhancer for hydrophilic drugs.The treatment of abnormal scar tissue remains a great challenge in clinical medicine since few carriers can penetrate scar tissue to enhance the biological function of the loaded drugs. Developing a transdermal carrier that can be loaded with drugs with distinct properties and penetrate scar tissue is critical for improving the efficacy of abnormal scar treatment . SNF cou2 fractional laser-assisted drug-laden SNF delivery in a rabbit ear HS model. Our data indicated that the laser treatment group R6G@SNF exhibited better penetration in the peri-CZ compared to R6G on day 1, while R110@SNF exhibited less penetration in the peri-CZ. The mean fluorescence intensity of the CZ was dramatically high and virtually unchanged on days 1, 3 and 5 for the laser-treatment groups. For the groups without laser treatment, this fluorescence intensity was weak. We concluded that combining CO2 fractional laser irradiation with SNF drug delivery could improve the efficiency of R6G (hydrophilic drug) delivery within 1 day . In contrast, the penetration of R110 (hydrophobic drug) was limited in the peri-CZ. A major problem that hindered the long-time permeability of the drugs was rapid re-epithelialization. The major contribution of this study was combining CO2 fractional laser irradiation with SNF drug delivery to improve the efficiency of hydrophilic drug delivery, offering a novel idea for the topical administration of water-soluble drugs. To our knowledge, this is the first time that SNF and CO2 fractional laser irradiation have been combined to promote drug penetration.In summary, this study explored COAFLs: ablative fractional lasers; AFM: atomic force microscopy; CCK-8: cell counting kit 8; CLSM: confocal laser scanning microscopy; CZ: coagulation zone; 5-FU: 5-fluorouracil; HDF: human dermal fibroblasts; HKF: human keloid fibroblasts; HS: hypertrophic scar; MAZ: microscopic ablation zone; peri-CZ: peri-coagulation zone; R110: rhodamine 110; R6G: rhodamine 6G; SNF: silk nanofiber hydrogel.