Triangularia manubriata sp. Nov.: A Novel Fungal Species Belonging to the Family Podosporaceae Isolated from Soil in Korea.

The fungal strain designated as KNUF-21-020, belonging to the genus Triangularia, was isolated from a soil sample collected in the Chungnam province, Korea. Phylogenetic analyses based on the concatenated nucleotide sequences of internal transcribed spacer regions and partial sequences of large subunit rRNA, beta-tubulin, and RNA polymerase II subunit genes revealed that the strain was grouped in a clade with Triangularia species. However, it occupied a distinct phylogenetic position. We also observed morphological differences between strain KNUF-21-020 and closely related species. Here, we provided detailed descriptions, illustrations, and discussions regarding the morphological and phylogenetic analyses of the closely related species to support the novelty of this isolated species. The phylogenetic analyses and morphological observations indicate that the strain KNUF-21-020 represents a novel species in the genus Triangularia (family: Podosporaceae). We have designated this species as Triangularia manubriata sp. nov.

Ultrasmall Mn-doped iron oxide nanoparticles with dual hepatobiliary and renal clearances for T1 MR liver imaging.

Although magnetic nanoparticles demonstrate significant potential as magnetic resonance imaging (MRI) contrast agents, their negative contrasts, liver accumulation, and limited excretion hinder their application. Herein, we developed ultrasmall Mn-doped iron oxide nanoparticles (UMIOs) with distinct advantages as T1 MRI contrast agents. Exceptionally small particle sizes (ca. 2 nm) and magnetization values (5 emu gMn+Fe-1) of UMIOs provided optimal T1 contrast effects with an ideally low r2/r1 value of ∼1. Furthermore, the use of Mn as a dopant facilitated hepatocyte uptake of the particles, allowing liver imaging. In animal studies, UMIOs exhibited significantly enhanced contrasts for sequential T1 imaging of blood vessels and the liver, distinguishing them from conventional magnetic nanoparticles. UMIOs were systematically cleared via dual hepatobiliary and renal excretion pathways, highlighting their safety profile. These characteristics imply substantial potential of UMIOs as T1 contrast agents for the accurate diagnosis of liver diseases.

Role of Kir4.1 Channels in Aminoglycoside-Induced Ototoxicity of Hair Cells.

The Kir4.1 channel, an inwardly rectifying potassium ion (K+) channel, is located in the hair cells of the organ of Corti as well as the intermediate cells of the stria vascularis. The Kir4.1 channel has a crucial role in the generation of endolymphatic potential and maintenance of the resting membrane potential. However, the role and functions of the Kir4.1 channel in the progenitor remain undescribed. To observe the role of Kir4.1 in the progenitor treated with the one-shot ototoxic drugs (kanamycin and furosemide), we set the proper condition in culturing Immortomouse-derived HEI-OC1 cells to express the potassium-related channels well. And also, that was reproduced in mice experiments to show the important role of Kir4.1 in the survival of hair cells after treating the ototoxicity drugs. In our results, when kanamycin and furosemide drugs were cotreated with HEI-OC1 cells, the Kir4.1 channel did not change, but the expression levels of the NKCC1 cotransporter and KCNQ4 channel are decreased. This shows that inward and outward channels were blocked by the two drugs (kanamycin and furosemide). However, noteworthy here is that the expression level of Kir4.1 channel increased when kanamycin was treated alone. This shows that Kir4.1, an inwardly rectifying potassium channel, acts as an outward channel in place of the corresponding channel when the KCNQ4 channel, an outward channel, is blocked. These results suggest that the Kir4.1 channel has a role in maintaining K+ homeostasis in supporting cells, with K+ concentration compensator when the NKCC1 cotransporter and Kv7.4 (KCNQ4) channels are deficient.

Sustainable Strategies for Synthesizing Lignin-Incorporated Bio-Based Waterborne Polyurethane with Tunable Characteristics.

In this study, we introduce a novel approach for synthesizing lignin-incorporated castor-oil-based cationic waterborne polyurethane (CWPU-LX), diverging significantly from conventional waterborne polyurethane dispersion synthesis methods. Our innovative method efficiently reduces the required solvent quantity for CWPU-LX synthesis to approximately 50% of that employed in traditional WBPU experimental procedures. By incorporating lignin into the polyurethane matrix using this efficient and reduced-solvent method, CWPU-LX demonstrates enhanced properties, rendering it a promising material for diverse applications. Dynamic interactions between lignin and polyurethane molecules contribute to improved mechanical properties, enhanced thermal stability, and increased solvent resistance. Dynamic interactions between lignin and polyurethane molecules contribute to improved tensile strength, up to 250% compared to CWPU samples. Furthermore, the inclusion of lignin enhanced thermal stability, showcasing a 4.6% increase in thermal decomposition temperature compared to conventional samples and increased solvent resistance to ethanol. Moreover, CWPU-LX exhibits desirable characteristics such as protection against ultraviolet light and antibacterial properties. These unique properties can be attributed to the presence of the polyphenolic group and the three-dimensional structure of lignin, further highlighting the versatility and potential of this material in various application domains. The integration of lignin, a renewable and abundant resource, into CWPU-LX exemplifies the commitment to environmentally conscious practices and underscores the significance of greener materials in achieving a more sustainable future.

D-Galactose and Hypoxia Induce the Early Onset of Age-Related Hearing Loss Deterioration in a Mouse Model.

BACKGROUND: We previously showed that aging accelerates after 3 months of exposure to hypoxia and environmental change but not genetic modifications. Here, we aimed to simply induce early-onset age-related hearing loss within a short period based on our previous method. METHODS: We randomly divided 16 C57BL/6 mice into four groups that were maintained under conditions of normoxia and hypoxia with or without injected D-galactose for 2 months. Deteriorated hearing, the expression of age-related factors, and oxidative stress responses were detected using the click and tone burst auditory brainstem response test, reverse transcription-polymerase chain reaction, and by measuring superoxide dismutase (SOD). RESULTS: The group maintained under hypoxia combined with D-galactose lost hearing particularly at 24 Hz and 32 Hz at 6 weeks compared with the other groups. Aging-related factors were also significantly decreased in the hypoxia and D-galactose groups. However, SOD levels did not significantly differ among the groups. CONCLUSION: Age-related hearing loss is an environmental disorder induced by chronic oxidative stress associated with genetic backgrounds. Our findings suggested that D-galactose and hypoxia can induce the phenotypes of age-related hearing loss and aging-associated molecules in a murine model within a short time with environmental stimulation alone.

Colorimetric mercury detection with enhanced sensitivity using magnetic-Au hybrid nanoparticles.

Due to the neural toxicity of mercury, there is a need for the development of on-site detection systems for Hg2+ monitoring. To this end, a new colorimetric mercury detection probe, Fe3O4@SiO2@Au (magnetic-Au; Mag-Au) hybrid nanoparticles, has been developed. The Au on the surface of Mag-Au is an indicator of Hg2+, which forms an AuHg alloy (amalgam) on their surface (Mag-Au@Hg), with excellent peroxidase-like activity. The oxidation of 3,3',5,5'-tetramethylbenzidine by Mag-Au@Hg resulted in a color change of the indicator solution, which was enhanced with increasing Hg2+ concentration. Mag-Au can be used to detect Hg2+ at nanomolar concentrations. Additionally, magnetic separation can be used to easily purify and concentrate the Mag-Au@Hg from samples, and thus avoid interference from unwanted residues or colored samples. The feasibility of Mag-Au for Hg2+ detection was tested with an artificial urine solution and it can be used to detect Hg2+ in various real samples, such as river water, seawater, food, and biological samples.

Enhancing catalytic efficiency of carbon dots by modulating their Mn doping and chemical structure with metal salts.

Nanozymes are emerging materials in various fields owing to their advantages over natural enzymes, such as controllable and facile synthesis, tunability in catalytic activities, cost-effectiveness, and high stability under stringent conditions. In this study, the effect of metal salts on the formation and catalytic activity of carbon dots (CDs), a promising nanozyme, is demonstrated. By introducing Mn sources that possess different counter anions, the chemical structure and composition of the CDs produced are affected, thereby influencing their enzymatic activities. The synergistic catalytic effect of the Mn and N-doped CDs (Mn&N-CDs) is induced by effective metal doping in the carbogenic domain and a high proportion of graphitic and pyridinic N. This highly enhanced catalytic effect of Mn&N-CDs allows them to respond sensitively to the interference factors of enzymatic reactions. Consequently, ascorbic acid, which is an essential nutrient for maintaining our health and is a reactive oxygen scavenger, can be successfully monitored using color change by forming oxidized 3,3',5,5'-tetramethylbenzidine with H2O2 and Mn&N-CDs. This study provides a basic understanding of the formation of CDs and how their catalytic properties can be controlled by the addition of different metal sources, thereby providing guidelines for the development of CDs for industrial applications.

Protection of Hearing Loss in Ototoxic Mouse Model Through SPIONs and Dexamethasone-Loaded PLGA Nanoparticle Delivery by Magnetic Attraction.

Background: Ototoxicity currently has no available treatment other than medication withdrawal as soon as toxicity is suspected. The human inner ear organs have little potential for regeneration; thus, ototoxicity-induced hair cell injury is deemed permanent. Dexamethasone (Dexa) is a synthetic steroid analog that has significant potential for otoprotection in the treatment of various inner ear diseases; however, its low absorption into the inner ear prevents significant recovery of function. Nanoparticles facilitate targeted drug delivery, stabilize drug release, and increase half-life of the drug. Methods: This study aimed to develop poly(lactic-co-glycolic acid) (PLGA) nanoparticles loaded superparamagnetic iron oxide nanoparticles (SPIONs) and Dexa (PSD-NPs) to control localized drug delivery by magnetic attraction in the treatment of ototoxicity-induced hearing loss. PSD-NPs and without SPIONs (PD-NPs) were prepared using a nanoprecipitation method. Results: Using an inner ear simulating system, we confirmed that PSD-NPs has an otoprotective effect in organotypic culture that is enhanced by magnetic attraction. PSD-NPs delivered via intrabullar injection in a magnetic field penetrated the inner ear and prevented hearing loss progression to a greater degree than equivalent doses of Dexa or PSD-NPs alone (day 28: ototoxic: 80.0 ± 0.0 dB; Dexa 100: 60.0 ± 15.5 dB; PSD 100: 50.0 ± 8.2 dB; PSD 100 with magnet: 22.5 ± 5.0 dB; P < 0.05). The protective effects were confirmed in various in vivo and in vitro models of ototoxicity. Conclusion: Our findings suggest that SPIONs with Dexa and magnetic field application prevent the progression of ototoxicity-induced hearing loss through anti-apoptotic mechanisms in the inner ear.

Mag-spinner: a next-generation Facile, Affordable, Simple, and porTable (FAST) magnetic separation system.

Mag-spinner, a system in which magnets are combined with a spinner system, is a new type of magnetic separation system for the preprocessing of biological and medical samples. Interference by undesired components restricts the detection accuracy and efficiency. Thus, the development of appropriate separation techniques is required for better detection of the desired targets, to enrich the target analytes and remove the undesired components. The strong response of iron oxide nanoclusters can successfully capture the targets quickly and with high efficiency. As a result, cancer cells can be effectively separated from blood using the developed mag-spinner system. Indeed, this system satisfies the requirements for desirable separation systems, namely (i) fast sorting rates, (ii) high separation efficiency, (iii) the ability to process native biological fluids, (iv) simple operating procedures, (v) low cost, (vi) operational convenience, and (vii) portability. Therefore, this system is widely applicable to sample preparation without limitations on place, cost, and equipment.

Zn-assisted modification of the chemical structure of N-doped carbon dots and their enhanced quantum yield and photostability.

This article presents the Zn-assisted synthesis of N-doped carbon dots (N-CDs) with an enhanced quantum yield (QY) and photostability. There have been intensive studies to improve or tune the optical properties of carbon dots (CDs) to meet the demand for luminescent materials in various fields, including energy conversion, photocatalysis, bioimaging, and phototherapy. For these applications, the photostability of the CDs is also a critical factor, but the related studies are relatively less common. The Zn-assisted N-CDs (denoted as Zn:N-CDs) obtained by the addition of Zn(OAc)2 to the precursors during the synthesis of N-CDs not only exhibited an enhanced quantum yield but also improved photostability compared to those of N-CDs. A comprehensive study of the chemical composition of Zn:N-CD and N-CD using X-ray photoelectron spectroscopy indicated a correlation between their chemical structure and photostability. Zn(OAc)2, which acts as a catalytic reagent, induced the modification of chemical structures at the edges of carbogenic sp2 domains, without being doped in N-CD, and the heteroatom-carbon bonds in Zn:N-CD seemed to be more resistant to light compared to those in N-CDs. The increased QY and photostability of Zn:N-CDs make them more suitable as an optical probe and they could be used in fingerprint identification. With Zn:N-CDs, the microstructure of fingerprints was confirmed clearly for a long duration effectively.

Hyperthermia Effect of Nanoclusters Governed by Interparticle Crystalline Structures.

Magnetic nanoparticles have an important role as heat generators in magnetic fluid hyperthermia, a type of next-generation cancer treatment. Despite various trials to improve the heat generation capability of magnetic nanoparticles, iron oxide nanoparticles are the only approved heat generators for clinical applications, which require a large injection dose due to their low hyperthermia efficiency. In this study, iron oxide nanoclusters (NCs) with a highly enhanced hyperthermia effect and adjustable size were synthesized through a facile and simple solvothermal method. Among the samples, the NCs with a size of 25 nm showed the highest hyperthermia efficiency. Differently sized NCs exhibit inconsistent interparticle crystalline alignments, which affect their magnetic properties (e.g., coercivity and saturation magnetization). As a result, the optimal NCs exhibited a significantly enhanced heat generation efficiency compared with that of isolated iron oxide nanoparticles (ca. 7 nm), and their hyperthermia effect on skin cancer cells was confirmed.

Highly Enhanced Enzymatic Activity of Mn-Induced Carbon Dots and Their Application as Colorimetric Sensor Probes.

Nanomaterial-based enzyme mimetics (nanozymes) have attracted significant interest because of their lower cost and higher stability compared to natural enzymes. In this study, we focused on improving the enzymatic properties of metal induced N-doped carbon dots (N-CDs), which are nanozymes of interest, and their applications for sensory systems. For this purpose, Mn(acetate)2 was introduced during the synthetic step of N-doped carbon dots, and its influence on the enzymatic properties of Mn-induced N-CDs (Mn:N-CDs) was investigated. Their chemical structure was analyzed through infrared spectroscopy and X-ray photoelectron spectrometry; the results suggest that Mn ions lead to the variation in the population of chemical bonding in Mn:N-CDs, whereas these ions were not incorporated into N-CD frameworks. This structural change improved the enzymatic properties of Mn:N-CDs with respect to those of N-CDs when the color change of a 3,3',5,5'-tetramethylbenzidine/H2O2 solution was examined in the presence of Mn:N-CDs and N-CDs. Based on this enhanced enzymatic property, a simple colorimetric system with Mn:N-CDs was used for the detection of γ-aminobutyric acid, which is an indicator of brain-related disease. Therefore, we believe that Mn:N-CDs will be an excellent enzymatic probe for the colorimetric sensor system.

Protective effect of MSC-derived exosomes against cisplatin-induced apoptosis via heat shock protein 70 in auditory explant model.

Therapeutics based on stem cell technology, including stem cell-derived exosomes, have emerged in recent years for the treatment of what were otherwise considered incurable diseases. In this study, we evaluated the efficacy of human MSC-derived exosomes for protection against cisplatin induced ototoxic hearing loss. Incubation of cochlear explants with MSC-derived exosomes prior to addition of cisplatin induced a reduction in cisplatin-induced drug toxicity in auditory hair cells but not when the exosomes were introduced simultaneously with or after cisplatin. The delivery of MSC-derived exosomes to cochlear explants was confirmed by the increasing protein levels of the exosome markers CD63 and HSP70 to reduce apoptosis. These results were consistent with those from a model in which MSC-derived exosomes protect auditory hair cells from cisplatin-induced drug toxicity in an ex vivo cochlear explant model and support future studies into the therapeutic benefits of stem cell-derived exosomes in clinical applications.

Biodistribution of poly clustered superparamagnetic iron oxide nanoparticle labeled mesenchymal stem cells in aminoglycoside induced ototoxic mouse model.

Recently, application of stem cell therapy in regenerative medicine has become an active field of study. Mesenchymal stem cells (MSCs) are known to have a strong ability for homing. MSCs labeled with superparamagnetic iron oxide nanoparticles (SPIONs) exhibit enhanced homing due to magnetic attraction. We have designed a SPION that has a cluster core of iron oxide-based nanoparticles coated with PLGA-Cy5.5. We optimized the nanoparticles for internalization to enable the transport of PCS nanoparticles through endocytosis into MSCs. The migration of magnetized MSCs with SPION by static magnets was seen in vitro. The auditory hair cells do not regenerate once damaged, ototoxic mouse model was generated by administration of kanamycin and furosemide. SPION labeled MSC's were administered through different injection routes in the ototoxic animal model. As result, the intratympanic administration group with magnet had the highest number of cells in the brain followed by the liver, cochlea, and kidney as compared to those in the control groups. The synthesized PCS (poly clustered superparamagnetic iron oxide) nanoparticles, together with MSCs, by magnetic attraction, could synergistically enhance stem cell delivery. The poly clustered superparamagnetic iron oxide nanoparticle labeled in the mesenchymal stem cells have increased the efficacy of homing of the MSC's to the target area by synergetic effect of magnetic attraction and chemotaxis (SDF-1/CXCR4 axis). This technique allows delivery of the stem cells to the areas with limited vasculatures. The nanoparticle in the biomedicine allows drug delivery, thus, the combination of nanomedicince together with the regenerative medicine will provide highly effective therapy.

Effect of heteroatoms on the optical properties and enzymatic activity of N-doped carbon dots.

Carbon dots (CDs) are attractive nanomaterials because of their facile synthesis, biocompatibility, superior physicochemical properties, and low cost of their precursors. Recent advances in CDs have particularly relied on the modulation of their properties by heteroatom doping (e.g., nitrogen). Although nitrogen-doped CDs (N-CDs) have attracted considerable attention owing to their different properties compared to those of the original CDs, the effects of the heteroatom content and types of bonding on the properties of N-doped CDs remain underexplored. In this work, we prepared N-CDs with controlled nitrogen contents, and fully examined their optical properties, enzymatic activity, and toxicity. We demonstrate that (i) the type of carbon-heteroatom bonding (i.e., carbon-oxygen and carbon-nitrogen bonds) can be altered by changing the ratio of carbon to heteroatom sources, and (ii) both the heteroatom content and the heteroatom-bonding character significantly influence the properties of the doped CDs. Notably, N-CDs exhibited higher quantum yields and peroxidase-like activities than the non-doped CDs. Furthermore, the negatively charged N-CDs exhibited negligible cytotoxicity. Such comprehensive investigations on the physicochemical properties of N-CDs are expected to guide the design of N-CDs for targeted applications.

Improvement of stem cell-derived exosome release efficiency by surface-modified nanoparticles.

BACKGROUND: Mesenchymal stem cells (MSCs) are pluripotent stromal cells that release extracellular vesicles (EVs). EVs contain various growth factors and antioxidants that can positively affect the surrounding cells. Nanoscale MSC-derived EVs, such as exosomes, have been developed as bio-stable nano-type materials. However, some issues, such as low yield and difficulty in quantification, limit their use. We hypothesized that enhancing exosome production using nanoparticles would stimulate the release of intracellular molecules. RESULTS: The aim of this study was to elucidate the molecular mechanisms of exosome generation by comparing the internalization of surface-modified, positively charged nanoparticles and exosome generation from MSCs. We determined that Rab7, a late endosome and auto-phagosome marker, was increased upon exosome expression and was associated with autophagosome formation. CONCLUSIONS: It was concluded that the nanoparticles we developed were transported to the lysosome by clathrin-mediated endocytosis. additionally, entered nanoparticles stimulated that autophagy related factors to release exosome from the MSC. MSC-derived exosomes using nanoparticles may increase exosome yield and enable the discovery of nanoparticle-induced genetic factors.

Induced Short-Term Hearing Loss due to Stimulation of Age-Related Factors by Intermittent Hypoxia, High-Fat Diet, and Galactose Injection.

Age-related hearing loss (ARHL) is the most common sensory disorder among the elderly, associated with aging and auditory hair cell death due to oxidative-stress-induced mitochondrial dysfunction. Although transgenic mice and long-term aging induction cultures have been used to study ARHL, there are currently no ARHL animal models that can be stimulated by intermittent environmental changes. In this study, an ARHL animal model was established by inducing continuous oxidative stress to promote short-term aging of cells, determined on the basis of expression of hearing-loss-induced phenotypes and aging-related factors. The incidence of hearing loss was significantly higher in dual- and triple-exposure conditions than in intermittent hypoxic conditions, high-fat diet (HFD), or d-galactose injection alone. Continuous oxidative stress and HFD accelerated cellular aging. An increase in Ucp2, usually expressed during mitochondrial dysfunction, was observed. Expression of Cdh23, Slc26a4, Kcnq4, Myo7a, and Myo6, which are ARHL-related factors, were modified by oxidative stress in the cells of the hearing organ. We found that intermittent hypoxia, HFD, and galactose injection accelerated cellular aging in the short term. Thus, we anticipate that the development of this hearing loss animal model, which reflects the effects of intermittent environmental changes, will benefit future research on ARHL.

Circulating microRNAs as potentially new diagnostic biomarkers of idiopathic sudden sensorineural hearing loss.

BACKGROUND: Early detection of inner ear cell damage can reduce the chances of permanent damage to hearing ability. However, current inner ear cell damage detection methods can detect damage only after the patient has lost hearing ability. MicroRNA expression levels in circulating systems are affected in diseases or conditions arising from the distant lesions. Therefore, detection of circulating microRNA expression levels could be one of the best ways to obtain information on inaccessible lesion sites. AIMS/OBJECTIVES: This study aims to establish a method for monitoring idiopathic sudden sensorineural hearing loss (ISSNHL) by analyzing circulating microRNA expression levels. 21 ISSNHL patients and 24 healthy controls were enrolled. MATERIAL AND METHODS: Real-time quantitative polymerase chain reaction was performed for detecting expression levels of circulating microRNAs. RESULTS: Among eight circulating microRNAs, expression levels of five circulating microRNAs significantly differed between ISSNHL patients and healthy controls. circulating microRNA expression levels correlates with treatment outcomes and hearing ability. CONCLUSIONS AND SIGNIFICANCE: Using methods combining the evaluation of miR-183, miR-210, miR-18b, and miR-23a cut-off values identified in ISSNHL patients and healthy controls during receiver operating characteristic curve analysis, sensitivity and specificity of 80.95% (17/21) and 87.50% (21/24) were obtained, respectively.

Endocytic trafficking of polymeric clustered superparamagnetic iron oxide nanoparticles in mesenchymal stem cells.

The technology of directing nanoparticles to specific locations in the body continues to be an area of great interest in a myriad of research fields. In the present study, we have developed nanoparticles and a method that allows the nanoparticles to move to specific sites by simultaneously utilizing the homing ability and magnetism of stem cells. Polymeric clustered SPIO (PCS) nanoparticles are composed of a superparamagnetic iron oxide nanoparticle (SPION) cluster core coated with poly lactic-co-glycolic acid (PLGA) and labeled with the fluorescent dye Cy5.5 for tracking. PCS is designed to be internalized by stem cells via endocytosis and then moved to the desired subcellular location through magnetism. Here, we investigated the interactions between SPIONs and mesenchymal stem cells (MSCs), including their absorption mechanism and subcellular localization. Exposure to the nanoparticles at 40 µg/mL for over 96 h did not affect cell survival or differentiation. We used a variety of endocytosis inhibitors and identified the potential cellular internalization pathway of SPIONs to be clathrin-mediated endocytosis. Antibodies to organelles were used to accumulate lysosomes through early and late endosomes. PCS at 40 µg/mL was internalized and stored without significant deleterious effects on stem cells, indicating that MSCs can act as an effective nanoparticle carrier. These findings also demonstrate the successful localization of the novel particles using magnetic attraction.

Non-invasive postoperative monitoring of pedicled rat skin flap using laser speckle contrast imaging.

PURPOSE: This study used non-invasive laser speckle contrast imaging (LSCI) modality to evaluate the blood flow changes in murine flap model and to investigate the clinical feasibility of the LSCI for postoperative monitoring. METHODS: Ten of 6-8 weeks old Spraque-Dawley rats with superficial inferior epigastric vessel based pedicled skin flaps were used in this experiment. The color changes of skin flap were evaluated by naked eyes and the LSCI modality 6, 24, and 48 h after surgery. RESULTS: In vessel ligated region of skin flap, skin color began to change to a bluish color immediately postoperatively. At 24 h postoperatively, skin necrosis was detectable with the naked eye and total necrosis occurred at 48 h postoperatively. Changes in laser speckle signal were consistent with changes observed with the naked eye, and blood flow index also presented significant differences between the ligated and non-ligated region. CONCLUSION: These correlated laser speckle signal patterns suggest that non-invasive monitoring of perfusion by LSCI is a useful technology that may be used to identify the ischemic skin flap.