The correct classification of salivary gland pathologies is crucial for choosing a treatment method and determining the prognosis. Better outcomes are now achievable thanks to the introduction of new therapy approaches, such as targeted therapies for malignant salivary gland tumors. To apply these in clinical routine, a clear classification of the lesions is required.The following review examines all changes from the first World Health Organization (WHO) Classification of salivary gland pathologies from 1972 to fifth edition from 2022. Possible developments in the diagnosis and classification of salivary gland pathology are also presented.The current WHO classification is the fifth edition. With the development of new diagnostic methods, based on genetic alterations, it provides insight into the molecular basis of lesions. This has resulted in the evolution of classification, introduction of new entities and reclassification of existing ones.Genetic alterations will become increasingly more significant in the identification of salivary gland pathologies in the future. These alterations will be helpful as prognostic and predictive biomarkers, and may also serve as targets for anti-cancer therapies. Salivary gland pathologies are a range of diverse diseases, therefore, classification is challenging. Moreover, developments in diagnostic methods, particularly at the molecular level, are allowing the discovery of novel subtypes of known diseases, that restrict the proper classification.The first edition of the WHO Histologic Classification of Salivary Gland Tumours  was publThe correct classification of a patient’s disease is crucial for choosing a treatment method and determining the prognosis. In the future, the development of modern treatment methods, including targeted therapies in management of malignant salivary glands tumours , would pThe first edition of the WHO classification of salivary gland diseases distinguished five carcinomas and two tumours . In subsequent editions, the number of distinguished malignant salivary gland lesions increased. The 1991 classification proposed 18 primary carcinomas. The next edition from 2005 distinguished 24 malignant epithelial tumours. Contrary, the number of malignant epithelial tumours was reduced to 20 types of carcinomas in 2017 and in the following classification from 2022, 21 different malignant salivary gland pathologies were identified. The changes in classifications are shown in Fig. Some diagnoses have remained the same since 1972. These include adenoid cystic carcinoma and carcinoma in pleomorphic adenoma (or carcinoma ex pleomorphic adenoma). Mucoepidermoid and acinic cell carcinomas were initially classified as tumours of uncertain malignancy, but were recognized as malignant lesions in the second edition of the classification. Although there has not been a reclassification of these lesions over the years, it should be emphasized that the definitions of individual diagnoses have been updated. The first two editions of the Blue Book based the classification on histological features seen with conventional light microscopy. Immunocytochemistry was limited to specific cases . The keyEssential modifications have occurred in the classification of adenocarcinomas. The first edition of the WHO classification  did not Despite the development of diagnostic methods and increasingly precise requirements for classifying lesions into a specific type of carcinoma, there are still some difficulties in distinguishing between certain pathologies. Some of these are discussed in the following paragraphs.An example is the relation between intraductal papillary mucinous neoplasm (IPMN) and mucinous adenocarcinoma. Mucinous adenocarcinoma (regardless of subtype) is characterized by a recurrent AKT1 p.E17K mutation , 9. The Intraductal carcinoma is characterized by proliferations entirely or predominantly intraductal. Some scientific reports state that invasive growth can appear in intraductal carcinoma, so it is not truly in-situ neoplasm and the name “intraductal” may not be correct –13.Oncocytic appearance is common in different salivary gland tumours. Lesions consisting entirely of oncocytes have been classified as oncocytic carcinoma. However, some studies at the molecular level indicate that these lesions should rather be classified as an oncocytic subtype of other carcinomas. To date, neither we have real evidence that purely oncocytic carcinoma exists, nor there have been discovered characteristic genetic alterations for this type of cancer , 14–16.The distinction between primary and secondary squamous cell carcinoma (SCC) of salivary gland still remains a diagnostic challenge. The majority of cases are metastatic tumours . The diaThe first edition of the WHO classification of head and neck tumours  distinguSimilarly to the malignant lesions, the involvement of genetic differences is also emphasized among benign lesions. Although still none of the benign salivary gland lesions is defined by genetic alterations, specific molecular changes have been identified and may provide an aid to classification and serve as potential biomarkers in the future. The most important genetic alterations in benign salivary gland lesions are shown in Table Although the classification of benign salivary gland lesions does not pose as many problems as in case of malignancies, the relationship between pleomorphic adenoma and metastasizing pleomorphic adenoma has caused controversy over the last few classifications. Pleomorphic adenoma, also called benign mixed tumour, is found mostly in the parotid gland in third to sixth decade of life, and occurs more frequently in women , 22. It The merit of genetic studies is the manifestation of the neoplastic features of sclerosing polycystic adenoma. This lesion was first introduced in the fourth edition of the WHO classification in the non-neoplastic epithelial lesion category . HoweverOther lesions described in the WHO classification included secondary and unclassified tumours, soft tissue tumours, lymphomas and non-neoplastic epithelial lesions.Non-epithelial tumours were classified since the first edition of the Blue Book . StartinHematolymphoid tumours were firstly added to classification in second edition, and described as malignant lymphomas . There aFor the first time in the fourth edition of the Blue Book, a category of non-neoplastic epithelial lesions was introduced . The maiA summary of the changes in the classifications described above is shown in Fig. Proper diagnosis of salivary gland pathology allows us to make the right therapeutic decision and determine the patient's prognosis. The most common treatment for salivary gland tumours is surgical resection, and the extent of surgery is determined mainly by anatomical and clinical criteria, but for some lesions an accurate diagnosis should influence therapeutic decisions. In the case of pleomorphic adenoma, the risk of tumour recurrence is about 2–3% and is highest in the myxoid subtype, as well as in the presence of thickness and incompleteness of the tumour capsule, pseudopodia, and satellite nodules , 42. ForAccurate differentiation of lesions is also important in planning treatment of canalicular adenomas, which have been divided into five different diagnoses in the latest classification of pathology . CurrentThe biggest differences in patient prognosis and treatment standards are seen when comparing different types of malignancies. If there are no contraindications, surgery with total tumour excision is the treatment of choice, according to NCCN Guidelines . PostopeChanges in the latest WHO classification  allow a 177 Lutetium PSMA [The uncovered specific molecular characterization of salivary gland cancers subtypes provides potential for exact definition and diagnosis but also perspectives for development of personalized therapeutic strategies. The described genetic alterations are oftentimes targetable, thus recurrent and metastatic cancers patients are already encouraged to participate in clinical trials. Patients with adenoid cystic carcinoma and MYB overexpression are included in the ongoing MYPHISMO trial with novel vaccination approach, used synergistically with programmed cell death protein 1 (PD-1) inhibitors . In turnium PSMA . In vitrium PSMA  and the ium PSMA . Adenocaium PSMA .The rare incidence of other salivary gland cancers subtypes and even the lower rate of metastatic and recurrent cases are so far not conducive to inclusion in clinical trials on systemic therapies.Initial classifications of salivary gland pathologies focussed on conventional histopathological examination. The second edition of the WHO classification  recommenA decisive direction in the development of diagnosis and identification of pathologies was introduced in the fourth version of the Blue Book , when emIncreasing numbers of researchers are highlighting the importance of genetic alterations as biomarkers of salivary gland pathology. It has been suggested that the genetic changes have also prognostic and predictive potential , 65. AltRecently, the importance of fine needle aspiration (FNA) cytology in the diagnosis of salivary gland lesions has increased . AlthougSo far none of the WHO classification of salivary gland pathologies includes imaging findings in the diagnosis of the lesions. Radiological examinations are also under constant development, and the utility of new techniques in the diagnosis of salivary gland proliferative lesions has been confirmed in recent studies. The recent significant progress in improving ultrasound imaging with the introduction of new technological solutions as shear wave elastography (SWE) and contrast enhanced ultrasonography (CEUS) influence the preoperative diagnostic workup in salivary gland pathologies. The studies published so far confirm the increased value of SWE versus classic ultrasound in differentiating between the most common benign lesions, polymorphic adenoma and adenolymphoma , 76. AltDynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) could be useful for recognizing the principal types of salivary gland tumours. The study of Mungai et al.  affirms Another perspective for the development of salivary pathology diagnostics is artificial intelligence. The first paper on using machine learning to evaluate salivary gland lesions was published in 2010. Siebers et al.  evaluateSome studies used machine learning to distinguish benign and malignant lesions of the parotid glands based on CT scans –87 and MCorrect diagnosis of salivary gland lesion is essential in determining the treatment and prognosis of the patient. Over the last 50 years, there have been many changes in the classification of salivary gland pathologies and the definitions of several lesions known for years have been updated. The most recent changes concern predominantly genetic studies results, which are increasingly being used in lesion classification. In the future, the importance of genetic alterations in the diagnosis of salivary gland pathology will increase even more. These alterations will also be helpful as prognostic and predictive biomarkers, and may also serve as targets for anti-cancer therapies.


Imagine an expensive research and development meeting at a large company. The presenter: “We have our top people working on this. Our top people!” This is how we feel about the many recent breakthroughs in auditory cognitive neuroscience research. Researchers like Tim Griffiths, Robert Zatorre, Andrew Oxenham, and the other contributors to this Frontiers' Research Topic have shaped and advanced the field for years. This collection of ten short perspective papers aims to provide a readable overview of several current  topics in auditory cognitive neuroscience through the vantage point of some of the main actors. The papers are best enjoyed as a collection rather than independently because of the many interconnections between the topics they discuss, some of which we will point out here.Oxenham discusses recent developments and directions in the study of pitch coding and perception.We start with topic of processing and representation of critical auditory features. The mechanism of pitch perception is among the oldest such topics in hearing science, going back to Strutt . The braTrapeau et al. present evidence-based arguments to support the role of the temporal voice area in genuine voice processing.From pitch extraction is the extraction of voice features: Pascal Belin's discovery of the temporal voice area in 2001  auditory cortex needs to catch up to that of the visual cortex. The auditory core is much smaller than V1 and is divided into subfields, nested on the superior temporal gyrus. Several functional and anatomical markers have been discovered and allow some non-invasive access, for example, increased myelination  in the framework of spectrotemporal modulation processing. The paper discusses the importance of low-level differential sensitivity to acoustical features of communication sounds (bottom-up) and high-level modulation of asymmetries by learning, attention, or other top-down factors.We now turn to the organization of the auditory system. Scott and Jasmin discuss the origins and recent developments of the dual stream concept and its interaction with speech and voice processing of simultaneous talkers.A central concept of sensory processing in the cortex is that of partially segregated streams with different functions. This idea was initially conceived to explain different sensitivities, and latencies in cortical fields along the visual pathway (Schneider, McAlpine and de Hoz discuss how adaptation in such feedback pathways of the auditory system aids in adaptive en- and decoding of complex sounds by building a representation of their statistical structure at different time scales. Exploring these feedback loops at different granularities, from in vivo recording to human neuroimaging, may reveal the fundamental listening processes.The feedback or top-down auditory projections is another principle of brain organization with powerful implications. The cortico-fugal pathway, the thickest efferent projection in the human brain after the pyramidal tract, instructively illustrates this. Griffiths provides an overview of recent work in the lab on predicting speech-in-noise ability based on performance with non-speech material in basic auditory cognitive tests. Speech in noise perception is the most important human auditory capacity and a consistent problem for persons with hearing disorders. Such tests may reveal the basic auditory factors that determine speech-in-noise understanding and enable more robust, language-independent clinical diagnosis. Rönnberg et al. discusses the ongoing trend of including more cognitive factors in this effort to add to the classical models based on system identification approaches to peripheral hearing mechanisms. He proposes the Ease of Language Understanding model, which models complex interactions of cognitive modules, such as the different memory systems, lexical access, and predictive and postdictive processes. Such models help to understand the perceptual consequences of hearing disorders and mirror the trend to include cognitive factors in hearing aids and rehabilitation.Finally, we turn to topics in more applied auditory neuroscience. All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.


Plant fiber reinforced polymer matrix composites have attracted much attention in many industries due to their abundant resources, low cost, biodegradability, and lightweight properties. Compared with synthetic fibers, various plant fibers are easy to obtain and have different characteristics, making them a substitute for synthetic fiber composite materials. However, the aging phenomenon of composite materials has been a key issue that hinders development. In natural environments, moisture absorption performance leads to serious degradation of the mechanical properties of composite materials, which delays the use of composite materials in humid environments. Therefore, the effects of moisture absorption performance of plant fiber composite materials on their mechanical properties have been summarized in this article, as well as various treatment methods to reduce the water absorption of composite materials. In the past few decades, synthetic fiber  reinforced polymer matrix composites have been used in various industries. However, most synthetic fibers have non-renewable and non-biodegradable properties, which affect the natural environment and promote the gradual replacement of elements of synthetic fiber composites by plant fiber composites ,2. CompaAt present, researchers have not only conducted extensive research on the hygroscopic properties of plant fiber reinforced polymer matrix composites, but also reported the impact of hygroscopic properties on the mechanical properties of composite materials ,19. The W1 and W2 are the initial mass before soaking and the final mass after soaking, respectively.The mechanical properties and dimensional stability of plant fiber composite materials were found to be adversely affected due to prolonged exposure to humid environments . TherefoIn recent years, researchers have developed different models to characterize the water absorption behavior of composite materials, but the overall approach is to model by considering diffusion mechanisms. The diffusion of water molecules into composite materials can be explained according to the Fickian diffusion law. At room temperature, the water absorption process of composite materials is very similar to the Fickian diffusion process ,29,30,31T is time, The diffusion types of plant fiber composite materials are shown in The high water absorption behavior between plant fibers and the matrix is the main drawback of plant fibers and the main reason for the decrease in mechanical properties of composite materials. The hydrophilicity of plant fibers is due to the presence of hydrogen bonds between cell wall molecules in the fibers. When the fibers come into contact with water molecules, the hydrogen bonds break and—OH groups form new hydrogen bonds with water molecules. Therefore, it can be concluded that hydrophilic—OH groups are the main source of fiber water absorption, and reducing the water absorption behavior of composite materials requires the elimination of hydrophilic groups. Due to the differences in plant fibers, their internal structures are diverse, and their inherent hydrophilic chemical components make it easy for them to absorb water during the soaking process, with varying water saturation levels. Although the water absorption rate of polymer matrix in plant fiber composite materials is far lower than that of fibers, the diffusion of water molecules in the matrix still conforms to the Fickian diffusion law, and the hygroscopic properties also affect the mechanical properties of the polymer matrix. The mechanism of the effect of hygroscopic properties on the polymer matrix is as follows: (1) Water molecules enter the matrix, which forces the distance between polymer molecular chains to increase, causing the matrix to swell and the polymer to undergo plasticization; (2) As the environmental temperature increases, the relaxation of polymer molecular chains intensifies, which weakens the intermolecular forces and accelerates the formation of molecular gaps within the polymer. The ability of water molecules to diffuse towards the polymer matrix is enhanced, and the water absorption rate and saturation amount are both increased; (3) The diffusion of water molecules towards the polymer matrix results in the formation of new cracks within the matrix due to the osmotic pressure generated. The continuous diffusion and increase of cracks further increase the water absorption capacity of the polymer matrix; (4) High temperature water molecules undergo hydrolysis reactions with hydrophilic groups such as amide groups, ether bonds, and amine groups in the polymer matrix, resulting in chain segments and cross-linking. The soaking time of different polymer matrices in water plays a crucial role in achieving water absorption saturation, as shown in 3) has been used to treat the fibers and a mixed NaHCO3 and polylactic acid coating to improve the water resistance and mechanical properties of sisal fiber (SH) composite materials [3 modified composite materials and mixed modified composite materials was relatively rough and there were polymer matrix particles, indicating that the adhesive strength between the modified fibers and the polymer matrix interface increases and there was no fiber debonding phenomenon. Gupta, M.K. et al. [The hygroscopic properties of plant fiber composite materials have a negative impact on their mechanical properties, and their industrial application performance is not good. Therefore, it is necessary to study the mechanical properties of composite materials after water absorption. The influence of moisture absorption properties on mechanical properties of sisal fiber (SF) composite materials are shown in aterials . Researc. et al.  studied . et al.  used 90 . et al.  studied 3 improved the surface modification of plant fibers, resulting in a 7% increase in the tensile strength of the composite material after water absorption. Additionally, the combination of NaHCO3 and PLA coating improved the tensile strength of the composite material after water absorption by 9.14%.As shown in As shown in In order to address the adverse effects of natural fiber moisture absorption on the mechanical properties and dimensional stability of composite materials, researchers have modified fibers through various chemical and physical methods. The various methods to reduce the water absorption of composite materials are introduced in this section, such as fiber surface chemical treatment, the use of compatibilizers, fiber mixing, nanofillers, and polymer coatings.2O2 (hydrogen peroxide), and prepared modified and unmodified short sisal fiber reinforced polylactic acid (PLA). The composite material showed a sharp increase in the first 24 h, followed by a stable trend, and the water absorption rate of the composite material gradually increased with the increase of plant fiber content. Compared with the unmodified short sisal fiber composite material, the water absorption rate of the modified composite material was much lower.Alkali treatment is one of the most common chemical modification methods, that works mainly by breaking the plant fiber OH group and reacting with water molecules to remove it from the plant fiber structure. It can also eliminate hemicellulose, lignin, and wax in plant fibers. This method can increase the surface roughness of plant fibers and improve the adhesion strength between fibers and polymers to minimize the water absorption of composite materials. Gunti Rajesh et al.  modifiedBenzoyl chloride in benzoylation mainly reduces the hydrophilicity of plant fibers and enhances the adhesion strength between plant fibers and polymers, where hydroxyl groups remove water absorbing substances such as wax and lignin through the fiber surface. Sreekumar, P.A. et al.  used varSilane coupling agent modification is one of the most effective methods to improve the interfacial adhesion of plant fiber composite materials. Cui, Y.H. et al.  studied Maleic anhydride treatment of plant fibers mainly reacts with—OH groups present in the amorphous region of the fiber cell wall, and reduces the hydrophilicity of plant fibers by forming a long polymer chain coating. Maleic anhydride grafted polypropylene (MAPP) has been used to treat loofah fiber (LF)/polypropylene composite materials . The stu3C0) and hydroxyl group (-OH) of the fiber to eliminate water. Bledzki, A.K. et al. [Acetylation treatment is used to change the structure of fibers, and is also known as the esterification method. In acetylation treatment, a reaction occurs between the acetyl group  to modify short jute fibers and prepared short jute fiber reinforced polypropylene composites. The study found that the water absorption of the composite material significantly decreased after NaHCO3 modification of short jute fibers. Fiore, V. et al. [Roy, J.K. et al.  used sod. et al.  preparedFiber hybrid reinforced polymer composites are composite materials with two or more different fiber reinforced polymers. The main purpose of introducing another fiber type into a single composite material is to maintain the advantages of the two hybrid fibers and reduce the disadvantages of a single fiber. As shown in the As shown in Gunturu, B. et al.  studied Mixing natural fibers with synthetic fibers (such as glass fibers and carbon fibers) are an effective method to solve the water absorption of composite materials. Mixing carbon fibers and glass fibers into plant fibers can improve the water resistance and mechanical properties of composite materials ,61,62,63Some researchers have attempted to coat polymers on the surface of fibers to improve the water resistance of composite materials, enhance interfacial bonding ability, and reduce the water absorption of composite materials. Rodriguez et al.  used a cWu, Y. et al.  used theNanofillers such as nanoclay and nano silicon carbide can be added to natural fiber composite materials to improve the water absorption and mechanical properties of the composite material in humid environments. A nanoclay layer forms an impermeable medium to prevent the flow of water molecules, allowing for longer diffusion time of water molecules, and reduces the water absorption of the composite material. Azam, F.A. et al.  found th2/jute fiber/polypropylene composite through melt gel technology and compression molding, as shown in 2 on the moisture absorption and interface failure of the composite were studied. Through molecular dynamics simulation, it was found that SiO2 effectively protected the composite from the influence of water diffusion by strong molecular chain interlocking on the interface of jute fiber/polypropylene composite, The water resistance and dimensional stability of the composite material were improved, and the water absorption rate and thickness expansion rate of the composite material were reduced by 3.6% and 3.0%, respectively.Liu, Y. et al. prepared a SiOAs shown in (1)The water absorption of composite materials is affected by various parameters, such as the selection of polymers and plant fibers, the mass fraction of plant fibers, fiber modification methods, water absorption time, and environmental humidity. However, the main parameters are the selection of plant fibers, fiber mass fraction, and water absorption time. It was concluded that plant fiber and synthetic fiber hybrid, polymer coating, and tile process coating have significantly improved the water resistance of composite materials, based on the current research.(2)The modification of the plant fiber polymer interface has always been a hot topic in the research on composite materials. Currently, there are many modification methods emerging, but the key technology for modifying plant fiber by polymer coating and Walli process coating still lags behind that in foreign countries. In addition, using these methods alone to treat plant fiber and its composite material’s water resistance cannot achieve the optimal state, Therefore, mixing polymer coating and Vary process coating with other physical/chemical methods for modification is an important research direction for improving the water resistance of plant fiber composite materials in the future.(3)Whether plant fiber composite materials can replace synthetic fiber composite materials in certain fields mainly depends on the changes in mechanical properties of plant fiber composite materials after water absorption. The authors believe that modifying plant fibers through various physical/chemical methods and different preparation processes can improve the mechanical properties of composite materials, but it cannot improve the moisture absorption properties of composite materials any further, ultimately leading to poor mechanical properties after water absorption. Therefore, it is necessary to develop a universal and low-cost technology to jointly improve the mechanical properties and water resistance of composite materials.(4)The researchers have developed water molecule diffusion models to study the water absorption behavior of composite materials and explain the water resistance performance of different composite materials. Therefore, developing different models to study the water resistance performance of composite materials is also an important direction for research in the future. By developing different water absorption models to express the water absorption behavior of plant fiber composite materials and continuously promote the development of new molding and modification processes, efforts will be made to achieve the development of high intelligence and low production costs, thereby promoting the widespread application of plant fiber composite materials.


Antennae in crayfish are essential for gaining information about the local topography and localising food, chemicals, conspecifics or predator. There are still gaps in the research on the morphology of antennae in decapods compared to other arthropods.Procambarus virginalis, Mexican dwarf crayfish Cambarellus patzcuarensis, red swamp crayfish Procambarus clarkii, signal crayfish Pacifastacus leniusculus, common yabby Cherax destructor, and spiny-cheek crayfish Faxonius limosus to find their potential morphological differences.Biometrical and ultrastructural methods were applied using light and cryo-scanning electron microscopies to study the morphology of antennae in six different crayfish species, including marbled crayfish Significant differences in the antenna length, length and width of each segment to carapace length ratios, and the number of segments were found in the six crayfish species. The ultrastructure revealed differences in the distribution of sensory hairs on the antenna and the morphology of the antennal surface.The different morphology of antennae might reflect adaptation to the conditions of their specific habitats. In addition, results showed that a combination of differences in the morphological features and biometrical measurements of antennae could be used for the distinguishment of different studied crayfish species. Decapod crustaceans bear segmented sensory organs on the anterior part of their bodies called antennae . They arCrayfish are a diverse group of decapods that are distributed across the different freshwater ecosystems in the world . Currentetc., but there is scarce information regarding the fine structure of this segmented appendage in crayfish , signal crayfish Pacifastacus leniusculus (Astacidae) and common yabby Cherax destructor (Parastacidae) to find the potential morphological differences among species.Therefore, we applied light and cryo-scanning electron microscopies to investigate the biometrical and ultrastructural features of the antennae from six crayfish species inhabiting different habitats, including marbled crayfish All protocols and ethics for animal handling were approved by the national laws and regulations on animal welfare 246/1992, and the institutional animal care guidelines of the Faculty of Fisheries and Protection of Waters of the University of South Bohemia in České Budějovice. All efforts were made to reduce animal suffering. It should be noted that no animals were killed during our study.n = 10), red swamp crayfish , spiny-cheek crayfish , and common yabby , crayfish were obtained from the experimental culture of the Laboratory of freshwater ecosystems, Faculty of fisheries and protection of waters, Vodňany, Czech Republic. Mexican dwarf crayfish  were purchased from the pet shop. Signal crayfish  were caught from Křesánovský creek near Vimperk, Czech Republic. Crayfish were separately acclimatised in plastic boxes with 1 L of aerated aged tap water before the experiment. Prior to sampling, the animals were anaesthetised on ice for 10 min. Antenna samples were collected equally from both sexes of adult crayfish using scissors and used for biometrical and ultrastructural purposes.Marbled crayfish  equipped with a DP74 camera was applied. The carapace length, antenna length, length and width of each antennal segment were measured by ImageJ . Antenna total length, width and lengths of each antennal segment in the different species were adjusted by dividing into the carapace length to remove differences among the size of animals .post-hoc test was carried out to evaluate differences among species. The length and width of each segment to carapace ratios, and number of segments were not normally distributed and assessed using the non-parametric Kruskal-Wallis test with subsequent multiple comparisons post-hoc statistical analysis. For all statistical tests, p < 0.05 was considered significant. Data are expressed as the mean ± SEM.For statistical analysis, Kolmogorov–Smirnov and Levene’s tests were used for the normality and homogeneity of variance using Statistica 13 software . For normal data (antenna to carapace length ratio), the one-way analysis of variance (ANOVA) with subsequent Tukey’s Scanning electron microscopy (SEM) JEOL 7401F  equipped with a cryo-attachment CryoALTO 2500  was used to study the antennal ultrastructure and sensory hairs. The samples were placed on the stub using double copper tape and Tissue-Tek. The samples were frozen by plunging into a liquid nitrogen slush. After freezing, the samples were transferred to the high vacuum cryo-preparation chamber, where the surface of the samples were sublimated at a temperature of −98 °C for 2 min. After sublimation, the samples were coated with 5 nm of gold at −135 °C. The coated samples were inserted into the Field Emission Electron Microscope  precooled at −135 °C. Images were obtained using an Everhart-Thornley detector at an accelerating voltage of 1.5 kV using Gentle Beam (GB) low mode .p = 0.0005, df = 5) difference in the antenna to carapace length ratio among our studied species. While common yabby showed the longest antenna to carapace length ratio, the signal and Mexican dwarf crayfish had the lowest values of this ratio  difference among the number of segments of the antennae in different species. While common yabby showed the highest number of segments, the Mexican dwarf and spiny-cheek crayfish had the lowest number of segments  difference among the lengths of each antenna segment to carapace length ratio in the different species. The longest and shortest lengths of the lower, middle and upper parts of each antenna segment were recorded in Mexican dwarf and red swamp crayfish, respectively  difference among the widths of each antenna segment to carapace length ratio in the studied species. The width of each segment for all species decreased from the lower to the upper parts of the antenna. For the different parts of the antenna, the widest and narrowest were recorded in Mexican dwarf and marbled crayfish in the upper part, common yabby and marbled crayfish in the middle part, and signal and Mexican dwarf crayfish in the lower part, respectively (Our results showed a significant (ectively .The scanning electron microscope micrographs showed a general conserved pattern, including segmented organs carrying two forms of sensory hairs on the surface in studied crayfish species –9. SensoMostly sparse and short sensory hairs were observed in the antenna of signal crayfish, and the branched form of sensory hair was sparsely distributed. The surface of the antenna in signal crayfish was squamous –4C.In Mexican dwarf crayfish, we observed mostly the non-branched form of sensory hairs and rarely the branched form. In most cases, the sensory hairs were as long as the length of the next segment. The surface of the antenna in this species is relatively wavy and wrinkled –5C.In marbled crayfish, we observed short and sparse sensory hairs of both forms. The sensory hairs were not frequent and relatively short, and the surface of the antenna is relatively squamous –6C.In red swamp crayfish, both forms of branched and non-branched sensory hairs were visible. They extended distally as far as the middle of the next segment, and the surface of the antenna in this species was relatively smooth –7C.Our observations in spiny-cheek crayfish showed that most of the sensory hairs in this species are branched. The surface of the antenna is relatively squamous –8C.In common yabby, we observed that the frequency of hairs in each segment is higher, and they consist primarily of the long, branched form. The surface of the antenna in this species is not smooth and covered by tubercles. The density of sensory hair is high, and the length of them is extended into half to whole length of an antennal segment –9C.Pontastacus leptodactylus including smooth and feathered sensory hairs. It was shown that while smooth hairs react against the surrounding environment movement and touching, feathered hairs response to direct touch and displacement of main shaft by bending (In the present study, we have demonstrated a comparative morphology of the antennae and accompanied sensory hairs among different freshwater crayfish species. The results of our study showed a general conserved pattern of morphology in antennae and their sensory hairs for all studied species. Two forms of sensory hairs were observed on the crayfish antennae, including branched and non-branched types. Similar forms of sensory hairs are supported by the literature. Two morphologically different forms of sensory hairs were detected on the antennae and antennules of narrow-clawed crayfish  bending . It was  bending . Contrar bending  and comm bending  are not  bending , which a bending  or chemo bending . On the Similar morphology of sensory hairs that were observed on antenna, antennule , chelae Troglocambarus maclanei and Cherax acherontis probably use their dense setose maxillipeds to collect organic material from the water current as a food source (Faxonius virilis (Antennae and sensory hairs morphology vary among species that may reflect adaptation for necessary mechanosensory and chemosensory requirements in the specific environment . For exad source . In addi virilis .Hence, the differences observed in the morphology of sensory hairs among different crayfish species could be caused by different levels of stimuli that they encounter in their specific habitat based on biodiversity, water quality, food availability, the intensity of intra- and interspecific interactions and density of predators. But it is unknown if the differences are evolved by short-term exposures during an individual’s lifetime or by long-term existence in specific habitats/conditions after several generations. Common yabby inhabits dark and turbid waters. Therefore, their tactile senses are a great source of information about their environment that highlights the importance of the antennae and sensory hairs in the common yabby . The antEven if we do not know the hidden drivers, we can say that the distribution and quantity of sensory hairs with various functions may have a significant impact on how surrounding information is received and can therefore affect the final response as described in other arthropods such as insects . It woulDespite the similar general morphology in the antennae and sensory hairs, there are differences in ultrastructural and biometrical traits that may help in the distinguishment of the studied crayfish species. Two forms of sensory hairs can reflect different functions or different developmental stages of these appendages. The different morphology of antennae might reflect adaptation to the conditions of their specific habitats. Different morphology and density of sensory hairs could also be a mark of different levels of sensitivity to detect chemicals, prey, predators, conspecifics and other stimuli in their environments. Further studies are needed to compare the sensitivity of different crayfish species to the stimuli based on their different sensory hairs density and morphology to find the best model species candidate for the behavioural and toxicological studies.10.7717/peerj.15006/supp-1Supplemental Information 1Click here for additional data file.


Community Series in Ethics in Psychiatry and Psychotherapy (https://www.frontiersin.org/research-topics/11564/ethics-in-psychiatry-and-psychotherapy).A multidisciplinary group of scholars provided significant contributions to Volume I of the The positive response to that collection resulted in a call for a second volume. Volume II continues the substantive ethical scholarship of its predecessor. The authors of these four articles represent a variety of methods, national representation, and topics that display the diversity of academic work in Ethics in Psychiatry and Psychotherapy. Two of the articles cover the classic topics of decisional capacity and assertive care from new perspectives and two feature discussions of the contemporary issues related to palliative care psychiatry and abortion laws.Kane et al. fills a gap in the qualitative literature with their report of semi-structured interviews with consultation-liaison psychiatrists working in general medical hospitals in England, Scotland, and New Zealand. Participants were asked to describe their experience of complex decision-making capacity assessments that presented both ethical and clinical challenges. A unique feature of the article is the narrative from consulting psychiatrists of how they managed these challenges. Thematic analysis of the interviews found four main categories of difficulty: discerning whether the patient's decision reflects their authentic values or dimensions of psychiatric or medical illness; appropriately applying ethical principles to the assessment outcomes; eliding personal bias in assessment of decision-making capacity; and problems in the process of assessment. Contrary to presumptions that psychiatrists would manifest conceits of paternalism and certitude, the respondents expressed epistemic humility and awareness of their own limits. The overarching lesson learned from the study is salutary for educators and mental health practitioners alike: in unraveling difficult decision-making capacity cases, ethics and psychiatry are entrained and expertise in both strands is necessary to principled resolution.Decision-making capacity evaluations are among the most common requests consultation-liaison psychiatrists receive from inpatient medical and surgical teams . The empLiégeois, the issue of decision-making capacity is again a central factor. The composite case study illustrates the relational grounding of the conceptual analysis. The author formulates the foundational ethical tension in mental health as that between the professionals' duty to respect the autonomy of patients with mental illness and to simultaneously safeguard them from harm. He draws on his service on the ethics committee of a large mental health care organization in Belgium to operationalize his thinking. Liégeois proposes a dimensional framework in which the two constituents are the care-users measure of decision-making capacity and the strength and certitude of the care-professionals concern for serious and imminent harm. The juxtaposition of these two variables creates six levels of ethically justifiable assertive care when the care-user possess some degree of decision-making capacity and four other levels when they are incapacitated. While striving to bring a more objective lens to what are often viewed as subjective decisions, each level remains relationally contextualized to prioritize the dignity and self-determination of the care-user and to preserve the trustworthiness of the treatment alliance whenever possible. Not unlike the sliding scale of capacity determination (In the contribution on assertive care by mination , more asMoureau et al. apply the method of scoping review (The final two articles in this volume explore current issues in mental health ethics. g review  to surveg review . The autTobón et al. are a group of American scholars who examine the impact on women's mental and physical health of the 2022 U. S. Supreme Court decision in Dobbs v. Jackson that reversed the access to abortion granted under Roe v. Wade and the cascade of restrictive state laws passed in its wake.Tobón et al. conclude that beyond “the physical morbidity, the psychological sequelae of carrying a forced pregnancy to term will lead to an even greater burden of maternal mental illness, exacerbating the already existing maternal mental health crisis”.The article highlights the significance of the Supreme Court's ruling and state legislation in several ways. The authors provide a data-informed appraisal of its clinical, educational, societal, research, and policy implications. They underscore that these often draconian statues will have the most adverse effect on already disadvantaged populations widening socio-economic inequities and deepening health care disparities such as maternal and infant mortality. Ethics in Psychiatry and Psychotherapy demonstrates the international reach, expanding methodology, and academic rigor of this academic enterprise. We trust that readers will find the series informative and engaging.Together with the articles in Volume I, this collection of thoughtful contributions on CG wrote the first draft of the manuscript. RB and MT revised and finalized it. All authors contributed to the article and approved the submitted version.


In this article, we provide an overview of the progress of scientists working to improve the quality of life of cancer patients. Among the known methods, cancer treatment methods focusing on the synergistic action of nanoparticles and nanocomposites have been proposed and described. The application of composite systems will allow precise delivery of therapeutic agents to cancer cells without systemic toxicity. The nanosystems described could be used as a high-efficiency photothermal therapy system by exploiting the properties of the individual nanoparticle components, including their magnetic, photothermal, complex, and bioactive properties. By combining the advantages of the individual components, it is possible to obtain a product that would be effective in cancer treatment. The use of nanomaterials to produce both drug carriers and those active substances with a direct anti-cancer effect has been extensively discussed. In this section, attention is paid to metallic nanoparticles, metal oxides, magnetic nanoparticles, and others. The use of complex compounds in biomedicine is also described. A group of compounds showing significant potential in anti-cancer therapies are natural compounds, which have also been discussed.  Cancer has affected living organisms for more than 200 million years. There is evidence of cancer among the ancestors of modern humans. Unlike infectious diseases, parasitic diseases, and other environmental diseases, cancer is not caused by some entity foreign to the human body. In this case, the destruction factors are human cells that have been transformed into pathological organisms or tumour building blocks . CancersCancer is the leading cause of death worldwide, accounting for 9.6 million deaths in 2017 . AccordiDue to the increasing incidence of various types of cancer, numerous studies are being conducted with the intent to develop a drug or select a suitable treatment method ,6. HowevCurrently, there are four main methods of cancer treatment, which include surgical resection, chemotherapy, radiation therapy, and biological therapy. Surgical resection is one of the most common ways of treating cancerous tissues and organs. However, in addition to surgical intervention, there are many other ways to mitigate the effects of cancer. They are used when surgical resection is not possible due to difficult access to diseased tissue, the presence of multiple and extensive tumour clusters, or the patient’s preference . The mai2O2. ROS generated during this treatment can induce apoptosis or necrosis by damaging the cellular components of cancer cells . To enable or enhance these “dynamic therapies”, nanoparticles (NPs) with small sizes, large surface areas, and numerous surface defects are often used as therapeutic agents with catalytic activity to facilitate ROS generation Cl). The complex was found to be stable in organic solvents and in physiological fluids. It exhibits pronounced cytotoxic activity in vitro against human SH-SY5Y cells. It was found that the in vitro antitumour activity is apparently related to the level of internalization of the drug. It is noteworthy that [Ag(EIA)2]Cl does not react with several model proteins but is capable of binding the C-terminal dodecapeptide of thioredoxin reductase hTrxR(488-499) and strongly inhibiting the activity of this enzyme. Internalization involves covalent binding of one or two carbene ligands to the C-terminal dodecapeptide with simultaneous release of the silver cation [A study by Fabbrini et al. concerned a silver(I) N-heterocyclic carbene (NHC) complex of bis(1-(anthracen-9-ylmethyl)-3-ethylimidazol-2-ylidene)silver ([Ag(EIA)r cation .The work of El-Tabl et al. presents new organic-metallic complexes constructed from a metal (II) bound to the H2L ligand, i.e., 2-hydroxy-N′-((Z)-3-(hydroxyimino)-4-oxopentane-2-ylidene)benzohydrazide. The complexes produced were characterized using a distorted octahedral geometry. An assay of the cytotoxic activity of the ligand and its complex with the metal showed potent cytotoxic effects against human liver cancer growth (test on HepG2 cell lines) compared to the clinically used Sorafenib (Nexavar) .In Bearing in mind the well-being of oncological patients, as well as the dynamic progress in modern anti-cancer therapies, it is extremely important to search for new methods of cancer treatment. This review presents the most modern techniques for treating diseased tissues with the use of a new generation of equipment and materials. Undoubtedly, metal complexes, compounds of natural origin, and metallic and oxide nanoparticles have great potential in the delivery systems of active anti-cancer substances and have shown anti-cancer activity themselves.The use of photothermal therapy in cancer treatment seems to be an appropriate direction to reduce the development of diseases, but it is unknown how the increase in temperature and the formation of active forms affect the active substances deposited on the surface of photothermal carriers. It is still important to understand in detail the mechanisms affecting the stability and photothermal properties of nanocarriers , metal nanoparticles, and complexes of these metals, as at present, very little is still known on these issues. The task is to design potential substances with synergistic anti-cancer effects, resulting from photothermal and magnetic properties and the release of metal ions into the tumour cell environment. These substances should possess the lowest possible toxicity to healthy cells but high toxicity to cancer cells, resulting from high uptake of ions released in the tumour environment and the formation of highly reactive oxygen radicals in the tumour environment.In connection with the hypothesis raised in the paper about the possibility of obtaining a nanocostructure that allows precise delivery of therapeutic agents to cancer cells and that could be used as a system with high efficiency of photothermal therapy, the questions remain: How could the efficiency of thermal energy storage by double layered hydroxides in such a construct be achieved? Will adsorption of metallic nanoparticles on the surface of metal oxide-LDH junctions increase their potential effect on cancer cells? The combination of metal nanoparticles and complex compounds of these metals in the construct will make it possible to reduce the release of ions and their harmful effects on healthy cells. Will this affect the stability of the complexes? Will the nanocostructure obtained have a function in anti-cancer therapies, and will it not exhibit toxic properties against normal cells?However, such an approach to the problem posed has not yet been reported in the literature. Solving the presented problem and obtaining answers to the posed questions will contribute to progress in obtaining new materials and developing anti-cancer therapies. Knowledge of the mechanisms of the above-mentioned chemical processes may be important for controlling the properties of both nanoparticles  and metal ion complexes (ensuring their ionic stability) and their interaction with photothermal compounds of biological significance. NIR radiation can result in the modification of chemical bonds and changes in the oxidation degrees of individual components, which should be limited; hence, research on this topic to obtain detailed information on the underlying processes is needed.Combating cancer is one of the greatest challenges of civilization in the coming decades. In view of the projected increase in the incidence of cancer and the resulting consequences in terms of high mortality rates and serious social and economic consequences, research on the subject carries great importance. The primary goal accompanying the presented problem is to improve the quality of life of oncological patients by minimizing the side effects of the therapy undertaken. This is because targeted therapy and direct action of the anti-cancer agent at the lesion site reduce the systemic reaction that occurs when using, for example, chemotherapy. From an economic point of view, it should be noted that cancer diseases generate both direct costs (resulting from their diagnosis and treatment) and indirect costs (related to losses in production and service revenues). Exploring the proposed topic will help reduce these negative consequences of cancer.


Exudate, an asymptomatic yellow deposit on retina, is among the primary characteristics of background diabetic retinopathy. Background diabetic retinopathy is a retinopathy related to high blood sugar levels which slowly affects all the organs of the body. The early detection of exudates aids doctors in screening the patients suffering from background diabetic retinopathy. A computer-aided method proposed in the present work detects and then segments the exudates in the images of retina acquired using a digital fundus camera by (i) gradient method to trace the contour of exudates, (ii) marking the connected candidate pixels to remove false exudates pixels, and (iii) linking the edge pixels for the boundary extraction of exudates. The method is tested on 1307 retinal fundus images with varying characteristics. Six hundred and forty-nine images were acquired from hospital and the remaining 658 from open-source benchmark databases, namely, STARE, DRIVE MESSIDOR, DiaretDB1, and e-Ophtha. The exudates segmentation method proposed in this research work results in the retinal fundus image-based (i) accuracy of 98.04%, (ii) sensitivity of 95.345%, and (iii) specificity of 98.63%. The segmentation results for a number of exudates-based evaluations depict the average (i) accuracy of 95.68%, (ii) sensitivity of 93.44%, and (iii) specificity of 97.22%. The substantial combined performance at image and exudates-based evaluations proves the contribution of the proposed method in mass screening as well as treatment process of background diabetic retinopathy. Background diabetic retinopathy (BDR), a systemic disease and long-term diabetes complication, is among the leading sources of blindness worldwide . AccordiIn accordance with eye specialists, growth in the number and size of exudates is the primary symptom of DR and is used as an indication to identify the stage of DR . ExudateThe detection of seriousness of DR requires qualitative and quantitative study of changes in exudates. Accurate identification of hard exudates contour is used for the laser photocoagulation process followed for the treatment of BDR. Eye specialists use manual inspection techniques to identify exudates boundaries in retinal fundus images. Manual marking and subsequently segmentation, as well as the precision with which lesions are assessed and associated criteria, are all hugely dependent on the specialist's skill and expertise. As a result, the uncertainty arises from (i) determining the precise borders due to various shapes and intensities, which may contribute to confusion, and (ii) the probability of missing exudates of a few pixels. Furthermore, manually scanning each retinal fundus image becomes tedious resulting in fatigue. Besides that, many patients are unable to receive effective care due to high costs of tests and scarcity of specialists. Thus, accurate identification of hard exudates contours is one of the primary tasks required for the detection of DR and subsequently treating it. Therefore, it is necessary to develop an exudates identification system to aid eye specialists, which will be beneficial in lowering the expense of specialists and eliminating the ambiguity related to manual marking. Moreover, exudates identification is beneficial not only for diagnosis but also for recovery preparation. Therefore, a computer-aided exudates identification and segmentation method of exudates for the diagnosis of BDR has been proposed in this research work. The rest of the article is organized as follows. In order to evaluate the diagnostic capacity of the exudates identification and segmentation methods proposed in literature, two types of assessment criteria are used, namely, number of exudates-based assessment and retinal fundus image-based assessments .Number of exudates-based assessment: each spot of exudates in retinal fundus image comprises a number of pixels and single infected image may contain one or more than one spots. These spots are segmented by applying exudates identification and segmentation method on retinal fundus images. The assessment of the segmentation results is carried out by comparing results pixel-wise with the reference markings by specialists using mathematically significant performance measures. The assessment results on exudates-based assessment should depict better performance as precise segmentation is necessary for the treatment planning of BDR.Retinal fundus image-based assessment: the major goal of retinal image-based assessment is to identify the occurrence or nonoccurrence of exudates to check any incidence of BDR or not. The results are simply dependent on the occurrence or nonoccurrence of exudates in the fundus. Image-based assessment is mainly carried out by correctly identifying retinal fundus images with exudates and nonexudates. This type of assessment is mainly used for clinical purposes, as the precise number of pixels is not significant from screening standpoint.The image-based assessment method evaluates the method's ability to distinguish between images with the occurrence or nonoccurrence of exudates. However, if the method does not have the capacity to identify contours of exudates, it demonstrates weak performance. Thus, in order to evaluate the method, it is necessary to evaluate the pixel level segmentation results, but it will provide more precise results than image-based assessment. Therefore, both the assessment criteria are key part in judging in the diagnostic capability of the method as well in clinical systems.A number of computer-aided methods for the identification and then segmentation of hard exudates have been developed to aid ophthalmologists for the detection of DR –22. The Various morphology-based methods were proposed in literature to identify and then segment exudates from retinal fundus images. Walter et al. segmented exudates and other structures present in retinal images based on morphological operators . NumerouNumerous researchers proposed hybrid methods using varying types of supervised approaches to segment hard exudates effectively –17. ClusLately, deep learning approaches are being applied on retinal fundus images for the identification and segmentation of lesions present in the retina. The deep learning approaches have an advantage of automatic feature extraction on huge number of images. Recently, numerous methods were developed for the detection of exudates and also they achieved promising results –21. ManyThe proposed method includes the following three steps: initially the exudates regions are enhanced using enhancement algorithm and also by eliminating artifacts present in the images captured with fundus cameras; in the next step, candidate exudates pixels are detected by applying gradient and linking based method. Lastly, the proposed method was validated by medically significant performance measures such as sensitivity, specificity, and accuracy. To demonstrate the generalisation capacity of the proposed method and applicability for practical usage, the suggested technique is created and evaluated using retinal images obtained from eye hospital as well as four different open-source benchmark databases, each with individual features such as capture device, area acquired, angle of acquisition, and resolution. The exudates segmentation results attained are then utilized by the eye specialists for the recovery planning of BDR. To summarize, the designed approach is intended to assist eye specialists in finding the severity of the disease and deliver higher accuracy.Retinal fundus images database in the present research work is captured from two platforms: 649 retinal images from eye hospital and 658 retinal images from four open-source benchmark databases. The purpose of using these images from variety of platforms is to test the efficiency of the proposed exudates identification and segmentation method to be independent of different variations in images. Open-source benchmark datasets used are STARE, MESSIDOR, DIARETDB1, and e-Ophtha –33. The Between March 2019 and December 2019, 649 retinal images of various intensities, resolutions, contrast, and clarity were obtained from SGHS Hospital, Punjab, India. A total of 375 images comprise hard exudates, 119 with other pathological structures and the remaining with no pathology. These images were taken from Topcon fundus camera, which has the resolution of 3218 × 2138 at an angle of 45°. The reference markings of exudates were provided by the eye specialists. These annotations were used as reference to compute the performance of the proposed method. The clinical supervisor provided the ethical approval to carry out this scientific study.MESSIDOR dataset: MESSIDOR database comprises of 1200 retinal images acquired using Topcon camera with different resolutions at an angle of 45° . Four huDIARETDB1 dataset: it contains 89 images taken with a fundus camera at an angle of 50° . Eighty-STARE dataset: a total of 400 images comprise hard exudates and other pathological structures such as red lesions of various shapes and sizes . These ie-Ophtha EX dataset: it contains 82 images comprising 47 pathological and 35 nonpathological images . RetinalOpen-source benchmark datasets have varying aims, attributes, and degree of comprehensiveness as discussed below:Reference markings of exudates: open-source benchmark databases do not comprise exact exudates markings or these annotations are no accurate to compute the performance related to exudates-based assessment. As a result, reference markings of all the images from different datasets are created under the supervision of eye specialists to precisely analyse the performance of the proposed method.The proposed algorithm in this work, as depicted in To enhance the quality of retinal fundus images for the accurate identification of exudates, it is necessary to remove artifacts in retinal images. These artifacts appear in retinal images due to several aspects such as inappropriate lighting of retinal fundus image, blur, focus, etc. These factors highly contribute to degrading the quality of retinal images. Degraded images can further result in inaccurate detection of retinal diseases. Thus, enhancement of images prior to the identification of exudates is an essential step. The algorithm developed for the enhancement of retinal images comprises three steps: (i) multiplicative transformation for elimination of artifacts due to improper lighting, (ii) channel selection for maximum intensity selection, and (iii) Weierstrass transformation for elimination of blur.I  is considered as a function of lighting l  and reflectance components r  shown inDependency of many factors related to camera, field of view, and expertise of the clinician results in varying types of artifacts in retinal fundus images. These factors result in variation of intensity in varying parts of retinal fundus images. Therefore, in the present work, multiplicative transformation is applied to eliminate the variation of intensity range and enhancing contrast in different parts of retina. In the proposed method, the retinal fundus image FT [ln {l }] and FT [ln {r }] are the FT of lighting and reflective components, respectively.Lighting component of retinal images represents smooth areas corresponding to low frequency and reflectance components represent high frequency components. High frequency components represent the details present in retinal image and thus need to be preserved. Therefore, in order to separate varying frequency components, the Fourier transform (FT) is applied on the image as shown inM given inIn the next step, the Fourier transform of the retinal image is filtered using maximally flat magnitude filter IP.In the last step, the resultant image is then converted from the frequency domain by applying inverse Fourier transform resulting in preprocessed image. The image is finally termed as The preprocessed retinal image contains RGB components. Among them, the green component represents the maximum intensity with respect to other structures present in retinal images and is thus selected for further processing.IPE.The blur present in retinal fundus images has a distribution similar to Weierstrass transformation . Hence, Exudates contour identification is the initial and significant tasks in the precise tracing of exudates edges. In order to identify the exudates contour, the following steps are designed in the proposed work.PEH and IPEV, respectively. The edge images obtained for both directions are used to calculate amplitude IPEamp and angle IPEang.The enhanced image obtained in the previous step is first passed through the Sobel edge detection filter to calculate the intensity variation using Del operator in both directions of Cartesian coordinates IThe edge images obtained in both directions are examined to eliminate unnecessary pixels not corresponding to exudates edges. The elimination process is carried out by analysing the entire image to determine whether it belongs to maximum intensity in the neighbourhood or not. It is done by comparing the pixel under consideration with all the other pixels of the neighbourhood. If the intensity value of the pixel under consideration is highest, then the pixel is kept otherwise eliminated.IPEamp is threshold by choosing two set of threshold values, namely, higher and lower threshold values, respectively. If the intensity value of IPEamp is more than the higher threshold value, pixel is preserved and termed as confirmed exudates pixels. On the other hand, if the intensity value of IPEamp is less than the lower threshold value, pixel is termed as false exudate pixel and eliminated. Also, if the pixel intensity value is more than the lower threshold value and less than the higher threshold value, the pixel is termed as weak exudates pixel. The decision on such pixels is taken based on the fact of whether these pixels are connected to confirmed exudates pixels or not. If they are connected, then these will be preserved along with confirmed exudates pixels; otherwise they will be eliminated.The resultant image of the last stage contains candidate exudates pixels. However, the application of the last step may result in false exudates pixels due to variation of intensities and artifacts in retinal images. In order to segregate the false exudate pixels, a dynamic thresholding method is designed in this work. The amplitude image The edges obtained in the previous step are comprised of the candidate exudates pixels. The pixels obtained in the previous step are broken or not connected due the nature of filters used. However, in order to extract the precise boundary of exudates, it is significant to link the exudates pixels extracted in the last step. Therefore, the steps followed to extract the exudates are the following.The candidate exudates pixels forming the exudates edges determined in the last step appear to be broken parts of exudates contours. The true edge contours are connected pixels forming clusters. However, the pixels which are not in the vicinity of the clusters are not the true exudates pixels and are to be eliminated. These insignificant false pixels are eliminated with the help of morphology diminishing operators.s” determine whether the centre pixel is the corner point or not. If at least one of these values is high, i.e., 1, the centre pixel is considered to be the candidate edge pixel; otherwise it is eliminated.The exudates edges in retinal images obtained in the previous step occasionally form the closed connected structures. The precise contour of exudates can be obtained only by connecting the corner points of exudates accurately. Therefore, the corner points estimation is necessary to connect the exudates edges. These corner points are retrieved by traversing the 4 × 4 array in the eight directions, as shown below. The corner point pixel is assumed to be in the centre of this array. The array values denoted by “The corner points of exudates retrieved in the previous step are marked to connect the exudates pixels accurately. A same mark is allocated to the set of exudates pixels depicting similar characteristics. The breaks in the exudates structures are linked by assessing the spatial properties of the corner points.Examine the features of exudate pixels in the neighbourhood of each marked exudate pixel.Pixels having the similar features are connected, resulting in forming the contour of exudates.Exudates pixels are connected if the difference between magnitude and angle between pixels under consideration is less than the predetermined magnitude and angle threshold, respectively.Lastly, the contour of exudates is detected by connecting the segments of exudates identified. Exudates segments are connected by neighbourhood analysis method. The designed method comprises 3 steps:The proposed exudates identification and segmentation method is designed and realized in MATLAB version 2018a on a PC with Intel Core i7 processor. Retinal images from hospital and four open-source benchmark databases, as mentioned earlier, are utilized to enumerate the performance of the proposed computer-aided method. A series of experiments were conducted to adjust the threshold values range at different steps. The optimal intensity thresholds are the ones that achieve best sensitivity/specificity combination. Increase in sensitivity indicates the correct detection of bright as well as subtle exudates, whereas higher specificity shows that the method does not recognize a nonexudates pixel as an exudates pixel. A total of 1307 retinal images comprising 525 nonpathological, 623 with exudates, and 157 with other retinopathies with different characteristics such as colour, location, resolution, etc. are used in this work. The performance validation of the designed method is done by (i) visual evaluation of expert ophthalmologist and (ii) medically important statistical measures such as sensitivity, specificity, accuracy, and positive predictive value . These sFigures Performance evaluation of the proposed computer-aided method in terms of medically significant performance parameters at two levels, i.e., exudates-based and image-based, on 1307 retinal fundus images is tabulated in In this evaluation, medically significant statistical measures are computed by comparing the ground truths of exudates boundary pointed by the expert ophthalmologist with the exudates segmentation results at exudates-based level by the proposed method. In this evaluation, medically significant statistical measures are computed by comparing the number of images marked by expert ophthalmologist as pathological, i.e., images with the incidence of exudates, and normal images with those of the designed method results. The overall mean sensitivity, specificity, and accuracy of 95.345%, 98.63%, and 98.04%, respectively, for image-based validation demonstrate the ability of the designed computer-aided method to discriminate images based on the occurrence and nonoccurrence of the exudates in contrast to the methods reported in literature. However, the method proposed by  shows 10A new exudates segmentation computer-aided method for the detection of background diabetic retinopathy has been designed in the present research work. The designed computer-aided method efficiently segments exudates in images of retina as it involves the steps such as (i) enhancement of retinal fundus images, (ii) candidate exudates contour detection, and (iii) precise segmentation of exudates. The performance of the designed exudates segmentation computer-aided method has been validated on 1307 images of retina from two sources, namely, eye hospital and four open-source benchmark databases. Results of the visual evaluation of the expert ophthalmologist are supported by the results obtained by the medically significant performance parameters, namely, exudates-based evaluation and image-based evaluation. The exudates-based evaluation outcomes specify the high precision of the designed method in correct detection of exudates pixels with sensitivity of 93.44%. The exudates-based evaluation results thus reveal the capability of the designed method in treatment process of background diabetic retinopathy. The specificity of 97.22% shows the ability of the method in successfully differentiating the exudates pixels from the nonexudates pixels. In addition, the proposed method outperformed another recent exudates segmentation method in terms of image-based evaluation with the accuracy of 98.04%. Therefore, the proposed method shows its potential in mass screening of patients with background diabetic retinopathy. Lastly, it can be highlighted the proposed computer-aided method may be used to assist the expert ophthalmologists in the preliminary detection and diagnosis of background diabetic retinopathy.


Dear editors,Recently, two Cochrane reviews on narrow band imaging (NBI)  and blueWhile our disclosures of clinical and academic interest, lecturing and advising must be taken into account, we are convinced that the quality of TURBT needs improvement.Both Cochrane reviews use the same methods showing reduced risk of recurrence by 37% and 34% for NBI and BLC and of progression by 35% for BLC; no progression data were available on NBI.We esteem Cochrane and acknowledge the reviews; there is the impression of a less rigorous interpretation and less balanced discussion of the NBI compared to the BLC data.p values are missing; such is unexpected (Table Six RCTs were included in the NBI review . Three ted Table .Table 1IIn the BLC report , missingThe BLC report  emphasizDespite being out of scope, the NBI report  emphasizThe pivotal event for NMIBC is progression; the NBI report  does notIn conclusion, considering the high-quality standards of the Cochrane reviews, differences in rigor and criticism of the evaluation and interpretation of BLC and NBI are tangible. We suggest the said aspects to be considered.The Cochrane review showed that BLC-guided TURB may reduce recurrence rates; such improvement is needed . InterprBLC may guide management following TURBT ; adjuvan


This study explores the clusters of closed restaurants in Seoul in response to the COVID-19 pandemic using the relative risk surface (RRS). The RRS developed based on kernel density estimation provides alternative perspectives for finding the cluster by combining different control and case events. Specifically, the varying impacts on diverse types of restaurants are examined by comparing the densities of closed casual restaurants and cafes. The clusters of closed businesses following the COVID-19 outbreak are subsequently explored through a comparison of the densities of the closed businesses preceding the outbreak. Furthermore, this analysis estimates the clusters of declined commercial areas after the pandemic outbreak based on the comparison between the densities of opened and closed restaurants. Finally, the specific time and region of the clusters are explored using space–time RRS. The analysis results effectively demonstrate various aspects of the closed restaurant clusters. For example, in the central business areas, the densities of closed cafes have decreased after the pandemic outbreak, and the density of closed cafes is significantly higher than that of opened cafes. This study would contribute to the literature on spatial data analysis and urban policy support in response to future epidemics. The Coronavirus disease (COVID-19) has spread to an unprecedented pandemic worldwide. As of 2022, the COVID-19 pandemic has globally surpassed five hundred million of confirmed incidences. Korea has been also suffered from the pandemic with the first case reported in Daegu in February 2020. From the second half of 2020, Seoul became the epicenter of the pandemic in Korea with the highest number of confirmed.1. From February 29, 2020, to May 16, 2022, the measure included restrictions on business hours and private gatherings, and recommendations for telecommuting and non-face-to-face classes in schools. However, the measure has also resulted in a significant reduction in population movement, which could have negative impacts on the sales of local businesses4.The Korean government has implemented social distancing measures that is known to as the effective COVID-19 countermeasure to mitigate the spread of the pandemic and reduce the number of confirmed cases5. Continued decreases in sales could result in the closure of these small businesses, ultimately contributing to a decline in the region6 and reduced tax revenue. To prevent business closures, local governments may provide subsidies to affected business owners. Therefore, identifying specific clusters of business closures can provide base research for policymakers to develop effective policy interventions.Exploring clusters of business closings in difference perspectives  can facilitate the development of locally specific policy supports to assist the regions that are suffered from the social distance measures. In Seoul, small business owners that could not overcome decreases in sales due to prolong the social distancing policy comprise a significant proportion of the industries8. Previous studies have also applied KDE to explore clusters of COVID-19 incidences10, and moreover, space–time kernel density estimation (STKDE) has been utilized to identify clusters in both spatial and temporal dimension of events11. While both KDE and STKDE can effectively identify clusters, these clusters may reflect only the population at risk of events when the events are closely related to their population at risk12. For example, the incidence of infectious diseases in a region is generally related to the population size of that region.Kernel density estimation (KDE) has widely utilized in various fields for exploring spatial clusters of point events13. RRS can be extended to various pairs of case and control densities, providing additional insight into cluster exploration. For example, it can explore temporal increases and decreases in case densities compared to corresponding control densities14. RRS identifies clusters of closed businesses resulting from the COVID-19 pandemic by comparing the densities of closed businesses before and after the pandemic outbreak. Moreover, space–time RRS (STRRS) emphasizes the region and specific time of spatio-temporal clusters15. Although the clusters RSS finds may not directly result from the pandemic, they can provide valuable exploratory data analysis results facilitating further investigations.Relative risk surface (RRS) and its extensions provide an alternative method for discovering different types of clusters, which could be useful for exploring clusters of closed businesses after the COVID-19 pandemic. RRS was initially developed to find statistically significant spatial clusters by comparing the densities of point events to that of their population at riskThe purpose of this study is to explore clusters of closed businesses, with a focus on restaurants, in Seoul, Korea, following the COVID-19 pandemic. Given that the restaurant industry is a major sector in Seoul, this study extends RRS to explore various clusters of closed restaurant businesses. Specifically, this analysis compares the impacts on different types of restaurants and identify clusters of closed businesses following the COVID-19 outbreak by comparing the densities of closed businesses before and after the outbreak. Additionally, RRS estimates clusters of declined commercial areas after the outbreak by comparing the densities of opened and closed restaurants. Finally, this study uses STRRS to explore the specific time of the clusters.This study identifies clusters of restaurant closures in Seoul following the COVID-19 pandemic outbreak using RRS and its extensions. According to the Korean 2020 census, Seoul has a population of approximately 9.6 million residents and is considered the most densely populated city in Korea. In addition, Seoul had the highest number of confirmed COVID-19 cases in Korea following the outbreak. Given the high population density and incidence rates in Seoul, the COVID-19 pandemic and subsequent social distancing policies have had a significant impact on local businesses.3. Specifically, according to the 2020 business survey conducted by Statistics Korea, the restaurant and lodging sector comprise the second highest proportion of the industries, accounting for about 16% of the total number of businesses. Additionally, the Seoul Institute reports a substantial decrease of 11% in restaurant industry sales in Seoul following the outbreak.This study examines the impact of the COVID-19 pandemic on local businesses with a focus on the restaurant industry. This industry was selected for the following analysis due to its significant contribution to the overall economy in Seoul and the negative impacts that it has experienced from the pandemichttps://www.localdata.go.kr/main.do, accessed on 1 March 2023).The impact of the pandemic on restaurant businesses may vary based on the type of restaurant. This study categorizes restaurant businesses into two major types, casual restaurants  and cafes , based on the Korean food hygiene law. To create restaurant opening and closure events, data on dates and geographic coordinates are obtained from the registration information provided by local administrative agencies 12, but it can be extended to various pairs of case and control densities14. This study employs three different case and control pairs to broaden the understanding of the negative impacts of the COVID-19 pandemic on restaurants businesses. First, this study compares the densities of closed casual restaurants and cafes to estimate the varying impacts on different types of restaurant businesses. Second, the densities of closed businesses before (2018–2019) and after (2020–2021) the pandemic outbreak are compared to explore the changes in restaurant closure densities. Third, the comparison between the densities of open and closed restaurants is conducted to estimate the decline in commercial districts. Although comparing closed and ongoing restaurants that are currently operated may be more appropriate, this study chose to use newly opened restaurants, instead of ongoing restaurants, to emphasize the possibility of rehabilitation of restaurant businesses22.The selection of proper case and control events is a crucial step in RRS, which has the potential to detect extended clusters using various case and control combinations. RRS initially compares the densities of case events to that of their population at risk23:Both RRS and STRRS are utilized to conduct the third comparison, which involves comparing the densities of closed restaurants to those of opened restaurants. Similar to the relationship between RRS and KDE, STRRS is developed based on STKDE. STRRS compares STKDEs of control and case spatio-temporal events as follows24. The permutation test compares the densities of case and control events and assesses the statistical difference between them. Specifically, the test randomly shuffles case and control events, and RRS and STRRS are estimated based on the shuffled events. This procedure is repeated 999 times, and p-values are computed based on the estimated reference distribution of RRS and STRRS values. The significance level for each cell should be inflated to avoid the multiple-testing problem26. However, this requires extensive increases in the number of permutations, and thus, this study could not apply the multiple-testing corrections . The simulated values can also be utilized for a global clustering test by comparing the sums of squared values27, which assesses whether the densities of case events are consistent with those of control events across a study area.RRS and STRRS provide a statistically significant test for identifying spatio-temporal clusters based on the permutation test8. Generally, a larger bandwidth size generates a smoother surface, but it may not adequately capture local variations in the estimated density surface. In contrast, smaller bandwidth size produces a spiky surface, which makes it difficult to visualize the global trends of an event distribution. This study estimates the optimal bandwidth size using the plug-in method28 based on the pooled standard deviations of control and case events, while also adjusting the estimated size for improved intuitive understanding. For RRS and STRRS, using the same bandwidth size for case and control data is recommended to maintain the same spatial and temporal ranges29.The selection of bandwidth sizes and kernel functions plays a crucial role in both RRS and STRRS, similar in KDE and STKDE. The size of the bandwidth is the most critical component because it determines the degree of smoothness in the estimated density surfaces13. This study employs a Gaussian density function to highlight the general trend of closed restaurant businesses because this function generates smoother surfaces than other kernel functions 30. Additionally, this study addresses underestimations of RRS and STRRS at the spatio-temporal edges of the study areas in the following ways. Spatial edge effects are corrected using Berman & Diggle’s method31, and temporal edge effects are adjusted by creating a toroidal wrap32.The selection of kernel functions also influences the smoothness of estimated density surfaces, although this choice is less critical than the selection of bandwidth sizeThis section presents the analysis results of the closed restaurants  clusters in response to the COVID-19 pandemic. First, the study provides a general perspective on the spatio-temporal distribution of the number of closed restaurants using a histogram and KDE. Next, this study examines changes in the densities of closed restaurants after the pandemic outbreak using RRS. Second, this study estimates RRS clusters obtained from various pairs of case and control densities. Specifically, it examines the varying impacts on different types of restaurant businesses by comparing the densities of closed casual restaurants and cafes. Furthermore, changes in the densities of closed businesses before and after the outbreak are explored by comparing the densities of closed businesses before and after the outbreak. Finally, this study estimates declined commercial areas after the pandemic outbreak using RRS and STRRS by comparing the densities of closed restaurants to those opened restaurants.Figure Figure 33.The KDE results Fig. A, B demop-value of 0.307 with 999 permutations). The areas with line fill symbols indicate statistically significant local clusters of changes in closed restaurant densities based on the permutation test. Positive RRS values in C-III, C-IV, C-V, and C-VII with line fill symbols indicate significant increases in closed restaurant densities after the COVID-19 outbreak. This highlights the increase in C-III  due to social distancing policies. Negative RRS values in C-I, C-II, and C-VI indicate that the densities of closed restaurants have decreased, although they still show relatively high densities of closed restaurants after the outbreak. The significant area in C-I successfully captures the changes in its shapes of C-I . For instance, C-I and C-IV in Fig. The RRS analysis compares the densities between closed and opened casual restaurants to assess the impacts of the COVID-19 pandemic on local casual restaurant businesses. Although the total number of opened casual restaurants  in Seoul is greater than that of closed casual restaurants 23,941) during the pandemic period (2020–2021), the RRS result indicate statistically significant local variations in the impacts of the pandemic on casual restaurants . For example, in Fig. The RRS analysis also examines the impacts of the pandemic on cafes by comparing the densities of closed cafes to those of opened cafes Fig. B. Simila18, which specifically indicate significantly higher densities of closed restaurants than opened restaurants at the 1% significance level.By comparing the spatio-temporal densities of closed and opened restaurants, STRRS identifies regions and time periods of decline in restaurant businesses after the pandemic outbreak. This study estimates the optimal sizes of spatio-temporal bandwidths for STRRS using the plug-in method and adjusted them to 2000 m and 90 days for spatial and temporal bandwidths, respectively, for ease of interpretation. Figure The STRRS analysis compares the spatio-temporal densities of closed and opened casual restaurants to identify spatio-temporal clusters of closed restaurants Fig. A. The coFigure This study presents an effective exploration of spatio and spatio-temporal clusters of closed restaurants in Seoul after the COVID-19 pandemic and highlights the utility of RRS extensions for the cluster exploration. First, this study examines the spatial clusters of closed restaurants using KDE, but it fails to effectively identify clusters that deviate from random chances due to inadequate consideration of their underlying population at risk. Second, this study extends RRS and STRRS by incorporating different combinations of cases and control functions, e.g., types of restaurants, times of closures, and closed and opened restaurants. These extensions enable more comprehensive exploration of closed restaurant clusters.Performing RRS and STRRS with various controls helps to detect significant clusters of closed restaurants by providing additional insights of the clusters. Specifically, the extensions of RRS and STRRS yields the following findings. First, the impacts of the COVID-19 pandemic on local restaurant businesses vary according to the types of restaurants, i.e., casual restaurants and cafes, which suggests the need to examine pandemic impacts by restaurant type. Second, the RRS method allows for estimation of changes in the densities of closed restaurants by comparing their densities before and after the pandemic outbreak. The results reveal regions that have experienced increases in densities . Third, the RRS method, which compares densities between closed and opened restaurants, is useful for capturing declines in local restaurant businesses. This result helps to recognize spatial clusters where restaurant businesses have declined from those that have not been negatively impacted by the pandemic. Finally, the STRRS method, which compares spatio-temporal densities of closed and opened restaurants, effectively captures both spatio- and temporal clusters of closed restaurants.This study would contribute to the literature on spatial data analysis and urban policy support in response to future pandemics. This analysis extends RRS and STRRS to detect spatio-temporal clusters of closed restaurants that are deviated from random occurrences with various pairs of cases and control. The extensions also provide a more comprehensive exploration of the impacts of the pandemic on local businesses. Specifically, the study successfully explores the varying impacts of the pandemic on the types of restaurants and identifies changes in the densities of closed restaurants after the pandemic outbreak. Additionally, the study can identify the decline of restaurant businesses by comparing closed and opened restaurant densities and highlight specific spatio-temporal clusters of these declines. As restaurant businesses are a major industry in Seoul, estimating their damages and decline trends in response to infectious diseases can inform policymakers in preparing support plans.35. Additionally, estimating difference-in-difference by comparing the changes between 2019 and 2018 with those between 2021 and 2020 could be helpful to eliminate temporal trends36. Third, this study estimates spatio-temporal clusters under the first-order separable assumption between space and time, which have been widely utilized in space–time analyses38. However, an approach to accommodate interaction between space time need to be applied in future studies39. Fourth, the analysis results potentially include type I errors due to the multiple-testing problem, and the results can generally be corrected based on Bonferroni and false discovery adjustments25. However, the application of these existing statistical and spatial statistical 40 correction methods into raster-based analyses warrants through investigation in following studies. Finally, although permanent business closure is generally considered the most negative impact of the pandemic on local businesses, it does not occur immediately in response to the pandemic. Therefore, excluding temporary closings41 and examining business sales3 could be useful in further investigating the impacts of the pandemic.This study can be extended in the several ways. First, alternative control events could be applicable in RRS and STRRS. For instance, this study compares opened dates of restaurants to their closed dates because both specific dates with their spatial coordinates are easily represented in the form of 3-D point datasets. However, the numbers of operating restaurants could provide alternative control events. Second, this study primarily focuses on detecting clusters of closed restaurants, and did not investigated their main factors contributing to these clusters through confirmatory spatial data analyses (CSDA), such as spatial regressions. The clusters this study finds could not be generated by the pandemic, and thus, an extension to CSDA could provide insight into the factors contributing to these clustersSupplementary Figure 1.