Dear Editor,In the early 2000s, neglected tropical diseases (NTDs) was coined as it refers to a set of diseases that occur in tropical countries due to the distinct geographical landscape, and distribution of the disease differs based on poverty in the region. Thus, they flourish in rural areas, conflict zones, and hard-to-reach regions, where there is generally a lack of quality healthcare, access to clean water, overcrowding, and scarce sanitation that is further aggravated by fluctuating climatic conditions. Pakistan ranks in the top 10 for the highest number of NTD cases for eight out of 18 NTDs researched in a study on global burden of diseaseThe country faces an impending debt crisis and perennial trade imbalance on the economic front, combined with multiple internal and external conflicts rampant, causing 46% of the Pakistani population to fall below the poverty lineIn Pakistan, some efforts have been taken to control the spread of NTDs such as the International Trachoma Initiative (ITI). This initiative began in 2016 and in partnership with the Pakistan Ministry of Health, has started mass drug administration of azithromycin to treat trachomaEradication of these diseases can be achieved with an increase in monitoring, evaluation, and surveillance with proper diagnostic tools and actions to reduce transmission. Greater availability of health resources in Pakistan’s rural areas could reduce the health burden of these diseases greatlyNone.None.M.Y.E. conceived the concept of the paper. H.A., J.A.S., and A.S. wrote the first draft of the manuscript. H.A. and M.Y.E. edited the second draft. All authors read and approved the final draft.The authors declare that they have no financial conflict of interest with regard to the content of this report.None.Mohammad Yasir Essar.


Hypericum L. comprises about 500 species distributed almost worldwide. Research has mainly focused on H. perforatum with confirmed biological activity on the alleviation of depression symptoms, among others. The compounds responsible for such activity are considered naphthodianthrones and acylphloroglucinols. Other Hypericum species are less studied or not studied, and further research is needed to complete the characterization of the genus. In this study we evaluated the qualitative and quantitative phytochemical profile of nine Hypericum species native to Greece, namely H. perforatum, H. tetrapterum, H. perfoliatum, H. rumeliacum subsp. apollinis, H. vesiculosum, H. cycladicum, H. fragile, H. olympicum and H. delphicum. Qualitative analysis was performed using the LC/Q-TOF/HRMS technique, while quantitative data were calculated with the single point external standard method. Additionally, we estimated the antioxidant activity of the extracts using DPPH and ABTS assays. Three species endemic to Greece  were studied for the first time. Our results indicated that all studied species are rich in secondary metabolites, mainly of the flavonoids family, with strong antioxidant activity. Hypericum L. (Hypericaceae) includes about 500 species placed into 36 sections, and has an almost worldwide distribution in temperate, subtropical and mountainous tropical regions  conH. perforatum, in which hyperforin dominates). For the rest of the examined extracts, the flavonoid content of H. rumeliacum subsp. apollinis reaches 771.41 mg/g dry material. In this extract, the most abundant flavonoids  were myricitrin , kaempferol glucoside  and I3, II8 biapigenin . Napoli et al. (2018) H. perfor. (2020) , regardiH. perforatum showed the best antioxidant activity as far as the DPPH assay, followed by H. delphicum. For the ABTS assay, all the calculated IC50 values were lower than those calculated for the DPPH assay. H. olympicum presented an IC50 value not far from that of the standard antioxidant, Trolox. The extract of H. vesiculosum was the least active for both assays used. Numerous studies on the antioxidant activity of different Hypericum species have been published. Taken together, data extracted from these studies make it clear that all Hypericum species possess strong antioxidant activity [The importance of antioxidants is an issue intensely discussed due to their multiple health benefits. Their use is not restricted only to the pharmaceutical industry, but antioxidants are also added in several food and cosmetic products ,50. The activity ,53,54,55The antioxidant activity of a compound is dependent on its chemical structure . AmentofH. perfoliatum, H. vesiculosum, H. cycladicum, H. perforatum and H. delphicum. However, the H. cycladicum extract, apart from the above-mentioned family of biologically active compounds, also contains a great amount of chlorogenic acid, the highest in comparison with the rest of the extracts. Luteolin glucoside and kaempferol glucoside are present. Further, the sum of flavonoids is high. For the ABTS assay, the calculated IC50 value of H. olympicum extract is the lowest. Despite the sum of flavonoids presented in this extract not being the highest one, some secondary metabolites, in particular myricetin glycosides, are detected in considerable amount. Myricetin is a strong antioxidant compound [H. rumeliacum subsp. apollinis extract, although in minor quantity. Myricitrin was also identified in this extract at a significant amount. Also of note, the H. rumeliacum subsp. apollinis extract is the richest regarding the sum of flavonoids and the second one in abundance of chlorogenic acid. H. cycladicum is another species with interesting antioxidant activity. The presence of luteolin glucoside and the higher amount of chlorogenic acid and amentoflavone, respective to the other extracts, contribute to its activity.Differences in the antioxidant capacity of the extracts are observed, which is rather expected taking into account the qualitative and quantitative variation between the identified secondary metabolites. All the tested extracts possess a significant antioxidant activity. Five out of nine species contain secondary metabolites of the flavan-3-ols and proanthocyanidins family. Proanthocyanidins are polyphenolic compounds, formed by flavan-3-ols subunits bound with B or A linkages. Most common flavan-3-ols subunits are catechin, gallocatechin and their isomers, from which oligo- or polymeric proanthocyanidins can be formed, among which oligo- proanthocyanidins are reported to be more effective antioxidants . Literatcompound . Both glH. vesiculosum is higher, a better antioxidant activity was estimated in the case of the H. delphicum extract. The sum of flavonoids for the two extracts is close; however, the better antioxidant activity for H. delphicum might be attributed to myricetin glucoside and kaempferol rhamnoside, which are present at great amounts in this extract. Generally, flavonoids are found in plants mostly in their glycosylated form, in particular at positions 3 (O-glycosylation) and 7 (C-glycosylation). As has been suggested, glycosylated forms are less active in terms of antioxidant activity. In addition, the number and type of glycosidic moieties alter the antioxidant activity, with glucose being the sugar to cause the minimum interference [H. perfoliatum, H. perforatum, H. delphicum (only epicatechin was identified) and H. cycladicum (only kaempferol was identified). Taken together, data of the antioxidant activity highlight that qualitative and quantitative differences are crucial for the radical scavenging activity of an extract. Definitely, a synergistic effect of secondary metabolites, including flavonoids, proanthocyanidins, quinic and benzoic acid derivates, contributes to the overall results. Naphthodianthrones and acylphloroglucinols seem not to contribute to the antioxidant activity [Although the number of compounds identified in the extract of rference . Most ofactivity .Hypericum species native to Greece, among which the endemic species of H. cycladicum, H. fragile and H. delphicum are studied for the first time. Generally, most of the studied species are overlooked compared to H. perforatum. Therefore, the present study adds novel information on the phytochemical profile and the antioxidant activity of the genus Hypericum.This study evaluated the phytochemical profile of nine Different families of secondary metabolites were identified, all documented by the existing literature to have a significant biological activity; hence, the present study gives strong motivation for further research on the biological activity of more species of the genus that will create a new perspective for their use. Our results present some similarities compared to those species for which studies have been performed. Nevertheless, important differences are also mentioned that highlight the unique climate characteristics between the different locations where the plant is found. However, the high demand for herbal products, in combination with exogenous factors such as climate change, pose risks for medicinal plants. As mentioned by , most me


Rajeswari et al., RSC Adv., 2014, 4, 61525–61536. https://doi.org/10.1039/C4RA12207K.Retraction of ‘Investigation of anticorrosive, antibacterial and  RSC Advances article due to concerns with the reliability of the data in the published article.The Royal Society of Chemistry, with the agreement of the authors, hereby wholly retracts this The HRSEM image in Fig. 4c represents a scaled version of part of the image presented in Fig. 4e. However, both images are presented with the same scale bar and represent different samples.Repeating fragments can be observed in the images in Fig. 6a–f, indicating that the images have been manipulated.The optical images in Fig. 8a–c have been duplicated in other publications. The panel in Fig. 8a has been duplicated as Fig. 9a in ref. The authors informed the editor that the characterization of the original samples was outsourced, and they do not have the original raw data for the published results.Given the significance of the concerns about the validity of the data, and the lack of raw data, the findings presented in this paper are not reliable.Signed: D. Rajeswari, D. Gopi, S. Ramya and L. KavithaDate: 16th March 2023RSC AdvancesRetraction endorsed by Laura Fisher, Executive Editor,


Concrete, one of the most often-used building materials today, is the cornerstone of modern buildings all over the world, being used for foundations, pavements, building walls, architectural structures, highways, bridges, overpasses, and so on. Because of its adaptability, concrete may be found in practically every construction, in some form or another. Yet, the diverse nature of its components, their combinations, and their doses result in a very wide range of concrete kinds with varying properties. As a result, concrete is a material that is always evolving and is popular even now, especially when it comes to circular economy.Other ways of concrete manufacturing are now being researched to lessen or remove the limits of this material, which are connected to its brittleness and poor environmental effects. As a result, the development of engineering cementitious composites has resulted in a significant reduction in flexibility issues, while the introduction of new additives and the optimization of the manufacturing process has resulted in a significant reduction in the negative effects of virgin raw material exploitation. In-depth research is still required to optimize and increase the sustainability of these advanced engineering cementitious composites or alternative concretes.2-eq. In another study, K. Khan et al. [In this Special Issue (SI), state-of-the-art research and review articles on the emerging material systems for AM are collected, with a focus on the process–structure–properties relationships. In total, eleven research papers and six reviews have been collected. Considering the high interest in this field for finding alternatives for virgin raw materials, in the research article conducted by Lv, Z. et al. , an inten et al.  showed tn et al.  conducteIn the review articles published in this SI, A.T.M. Yin et al.  discusse


Rickettsia aeschlimannii infection is an emerging human tick-borne disease with only a few recorded cases. We reported a presumable autochthonous case of rickettsiosis in an Italian cattle breeder associated with a Hyalomma marginatum bite. Rickettsia aeschlimannii DNA was detected in both the tick specimen from the patient and the grazing cattle close to his farm.  Rickettsia aeschlimannii is an emerging human tick-borne pathogen responsible for typical signs of Mediterranean spotted fever (MSF), acute hepatitis without MSF-like signs, and fever with meningitis without inoculation eschar or cutaneous rash [Hyalomma marginatum ticks and characterized as a new member of Spotted Fever Group (SFG) rickettsiae [R. aeschlimannii was linked to a human infection case in a French traveler returning from Morocco [R. aeschlimannii are mostly linked to imported cases with only one autochthonous clinical case recorded in Italy [R. aeschlimannii DNA has been frequently detected in several species of ticks removed from mammals and birds [ous rash ,2,3,4. Tkettsiae . Subsequ Morocco  and it h Morocco ,8,9,10. nd birds ,12. We r17kDa) and the citrate synthase-encoding gene (gltA) from the specimen as described by Webb and Roux [Hyalomma marginatum marginatum (H. m. m.). Sequencing of the PCR products showed 100% identity with Rickettsia (R.) aeschlimannii. Due to the circumstances of the events two years earlier, it was impossible to determine the presence of R. aeschlimannii by molecular tests, rickettsia isolation or immunohistochemistry in the patient’s lesions. Unfortunately, despite our request for a serological test, the patient found the blood draw inappropriate. The cattle farmer, in fact, considered the molecular detection of the pathogen in the preserved tick sufficient to solve his diagnostic doubt.During May 2022, a 75-year-old cattle breeder, male, living in a rural area of Apulia region (Gargano promontory) and without travel history delivered to our laboratory one tick specimen to be analyzed for pathogens. The ectoparasite was stored in 70% ethanol in a plastic vial dated 27 September 2020. The late delivery of the sample was due to the COVID-19 restrictions and fear of the contagion at the time. Questioned, the patient referred that the ectoparasite had been manually removed from his foot during the requested medical consultation; in the occasion, a therapy with antibiotic ointment and amoxicillina 1000 mg daily was also prescribed. On the fourth day, despite the ongoing antibiotic treatment, fever (peak 38.2) and foot pain and swelling accompanied by itch appeared; in the following days, the tick-bite area became progressively darker. The patient did not remember the presence of a maculopapular skin rash associated with the fever but he reported  the development of an expanding dermal redness at the place of the tick bite. The subsequent therapy with doxycycline 200 mg daily for 14 days gradually led to remission of the referred symptoms. To prove the story, he also provided a series of photos documenting the developed lesions on the tick-bite site in the period October–November 2020; in addition, a more recent photo was also taken at that date . Due to and Roux  were sucH. marginatum, along with Ixodes ricinus, Dermacentor marginatus and Rhipicephalus sanguineous, is regarded as one of the most common tick species of medical interest. In Italy, the genus Hyalomma Koch, 1844 includes the species H. aegyptium, H. lusitanicum, H. detritum (formerly H. d. detritum and H. d. scupense subspecies), H. excavatum and H. marginatum (formerly H. m. marginatum and H. m. rufipes subspecies) [H. marginatum complex are characterized by a high ecological plasticity and they can be found in very different environments, including arid or humid pastures, in low, medium and high mountains or in the Mediterranean maquis. In Italy, adult and immature specimens of H. m. marginatum were historically reported in continental and insular regions, including the Apulia region [H. marginatum is an exophilic and ditropic tick with a cycle of two hosts where only the larvae and adults need to find a host. Adults usually feed on large mammals, mainly artiodactyls and perissodactyls, while the immature forms are commonly ectoparasites of lagomorphs, rodents and of a wide number of birds [H. marginatum is also a well-known vector and reservoir for various zoonotic and emerging pathogens, including Crimean–Congo hemorrhagic fever (CCHF) virus and R. aeschlimannii; both these pathogens can persist in the arthropod for their entire lifespan, and they can be also transmitted vertically to the next generation. The Mediterranean basin’s warmer parts, in fact, house an autochthonous population of H marginatum but the evidence of a trend towards a warmer climate in Europe and the probability of introduction of immature specimens by birds has increased the recognition of this tick species as a possible invasive species for the countries of northern Europe [H. marginatum is considered to be the main reservoir of R. aeschlimannii [H. m. m in a transstadial way. H. marginatum can transmit R. aeschlimannii to future generations through transovarian transmission, promoting the ability of this tick species to maintain and colonize new territories, hosts and environments. Birds, cattle, small ruminants, camels and rodents, although asymptomatic, could be the vertebrate reservoirs of the pathogen in infected areas. The ability of R. aeschlimannii to infect other extremely prolific tick and mammal species has been also reported. In Algeria, in fact, the DNA of R. aeschlimannii has been recently amplified in specimens of Rhipicephalus bursa (Anatolian brown tick) collected from cattle and in Tunisia the bacterium was isolated from wild rodents [R. aeschlimannii-infected tick specimen. After the H. marginatum bite, his foot quickly became so swollen that he could not wear shoes and the pain was so intense that it could not walk. The patient’s awe increased due to the onset of the fever and to the worsening of the skin lesion caused by the tick bite. He did not report the appearance of spots or a maculopapular rash. During this time, the clinician contacted by telephone after assessing the symptoms and history suspected a tick bite infection, such as Lyme disease or rickettsiosis.In the Mediterranean area, species) ,16. The a region ,18,19. Hof birds . The abiof birds . In all n Europe  with thelimannii ,23,24,25 rodents . In our tache noir fell and the wound improved gradually after receiving antibiotic treatment with doxycycline. The patient’s cousin took care of the cattle during his convalescence and did not report any health problems. Although the general condition of the patient improved quickly and the swelling and pain of the foot disappeared after about 7 days, at the site of the tick bite, the healing of the skin was very slow. About 600 days after the parasite removal, a skin area with abnormal pigmentation was still visible. The cattle examined were in good health and showed no signs of infectious disease. The IZSPB database consultation confirmed that, in addition to H. marginatum, several species of ticks have been previously collected in the investigated area, such as I. ricinus, D. marginatus, Rh. turanicus, Rh. sanguineus, Rh. bursa and Haemaphysalis inermis; furthermore, zoonotic tick-borne pathogens like Borrelia garinii and B. afzelii [R. helvetica, R. raoultii, R. slovaca, R. felis [R. monacensis had also been identified in questing ticks collected in the same area. The detection of these pathogens in competent vectors suggests that the number of human infections related to tick-borne diseases could be higher than the scarcely reported cases in the medical literature from Southern Italy, including the Gargano area. This is supplemented with the fact that the diagnosis of R. aeschlimannii infection may be extremely challenging; in fact, in addition to the MSF manifestations, characterized by eschar, rash, edema and partial necrosis surrounding the tick bite site, episodes may present with nonspecific clinical symptoms , without cutaneous signs, such as observed in the case of an infection acquired in Southern Italy [R. aeschlimannii could be the cause of necrosis, erythema, myalgia, edema, and partial necrosis surrounding the tick bite site with clinical manifestations analogous to those described in several infections with SFGR. The clinical signs illustrated in our study are very similar to a report case of an R. aeschlimannii infection in a woman in China [R. aeschlimannii in ticks around the patient’s home and they associated the human infection with the R. aeschlimannii detected in vectors. On the basis of the above, a diagnosis of rickettsiosis is still a challenge, especially when it is solely based on clinical manifestations. Since the development of the Weil-Felix test, serology has been the primary method for diagnosing rickettsial infections. Increased sensitivity and specificity have been achieved by advances in serological analysis, but several critical aspects still remain unsolved. Among these limits is the correct identification of species. R. rickettsii or R. conorii antigens are the only ones used in many commercial assays that use indirect immunofluorescence assays (IFAs). Discarding Rickettsia groups but not species can also be achieved by using an enzyme immunosorbent assay (ELISA) for IgM or IgG for SFG and Typhus Group rickettsial infections. An alternative technique that has been shown to be effective for the differentiation of antibodies against different Rickettsia species is the micro-IFA (MIF). The assay was able to discriminate specific antibodies for R. helvetica, R. raoultii and R. slovaca in samples of canine serum [R. aeschlimannii, R. conorii, R. africae, R. slovaca, R. helvetica and R. massiliae in human serum samples [17kDa and gltA genes allowed us to characterize the species detected as R. aeschlimannii. In addition, entomological findings show that the investigated area houses several specimens of H. m. m. that are potentially dangerous for human health, highlighting that the percentage of ticks collected and found to be infected was 100%. There is still a lack of information regarding the role of mammals or birds as potential reservoirs of zoonotic rickettsiae. Despite the identification of R. helvetica DNA in deer blood samples [R. aeschlimannii, in domestic and wild ruminants is still largely uninvestigated. In conclusion, due to the high prevalence of R. aeschlimannii in the examined ticks and technical difficulties of making a retrospective serological diagnosis of infection among the SFG rickettsiae, we suppose that there is a fairly plausible correlation between the removed tick and the clinical manifestation of the patient. The history and the clinical signs are highly presumptive of tick-borne rickettsiosis. Although the presence of R. aeschlimannii has been ascertained only in ticks, further investigations in vertebrate hosts should be recommended to identify the vertebrate reservoirs of R. aeschlimannii in the Mediterranean region. In sites where ticks are present, the differential diagnosis between the various cutaneous clinical findings has certainly been a medical challenge. Interestingly, there is the evidence that both COVID-19 (SARS-CoV-2) and rickettsial infection can have cutaneous involvement with eschar and necrotic lesions; in fact, both share a similar mechanism of infecting endothelial cells, resulting in vasculopathy lesions [Due to the lack of available serological tests in the medical laboratory and the severe restrictions associated with the SARS-CoV-2 infection, therapy was prescribed for suspected rickettsiosis. The  afzelii , R. helvR. felis  and R. mrn Italy , or may rn Italy . R. aescin China . In the ne serum  and R. a samples  and of c samples , the pre lesions . However


Conocybe belongs to the Bolbitiaceae. The morphological classification and molecular phylogenetics of Conocybe section Pilosellae are not in agreement. In this study, based on the specimens from China, we investigated the sect. Pilosellae and identified 17 species, including 7 new species: Conocybe pilosa, with a densely hairy pileus and stipe; C. reniformis, with reniform spores; C. ceracea, with waxy dehydration of the lamellae; C. muscicola, growing on moss; C. sinobispora, with two-spored basidia; C. hydrophila, with a hygrophanous pileus; C. rufostipes, growing on dung with a brown stipe; and C. pseudocrispa, one new record for China. A key was compiled for the sect. Pilosellae in China. Here, the sect. Pilosellae, and new species and records from China are morphologically described and illustrated. Maximum likelihood and Bayesian analyses were performed using a combined nuc rDNA internal transcribed spacer region (ITS) and nuc 28S rDNA (nrLSU), and translation elongation factor 1-alpha (tef1-α) dataset to reconstruct the relationships of this section. We found that the sect. Pilosellae was the basal clade of Conocybe, and its evolutionary features may shed light on the characteristics of Conocybe. By integrating morphological classification and phylogenetic analysis, we explored the possible phylogenetic relationships among the species of the sect. Pilosellae in China. Conocybe Fay  originates from the Greek words “konos” and “cybe”, meaning “conical head” and refers to its conical pileus .2.Conocybe singeriana Hauskn. [ Hauskn. ,18,56C. singeriana belongs to the sect. Pilosellae, it is molecularly classified under Conocybe Clade1 and should be excluded from the sect. Pilosellae.Collection: Mulun Township, Huanjiang Maonan Autonomous County, Hechi City, Guangxi Zhuang Autonomous Region, China. 9 April 2021, HMJAU64956. Although the caulocystidia morphology of Conocybe was a monophyletic group, whereas Pholiotina was a polyphyletic group. Moreover, the sect. Pilosellae was the basal section of Conocybe. This result has also been confirmed in the studies by Tóth et al. (2013) [Pilosellae is necessary because this section exhibits diverse forms of species evolution at the basal node of Conocybe  and the morphological classification of Hausknecht (2009) ,19, we i13 and th. (2013) . PiloselConocybe . For exa section ,57. C. iConocybe . MoreovePilosellae from China, attempted to align morphological classification characteristics with molecular systematics to determine the classification and evolutionary characteristics of this section. It was discovered that the shape of the pileus of the species in this section is predominantly conical, with campanulate and hemispherical shapes being less common, and subglobose being the least common. A hygrophanous pileus may be correlated with the presence or absence of striations. Most species in this section have a slightly hairy pileus, with only a few species having a highly hairy pileus. The edges of the lamellae are either smooth or serrated. The stipes of all species in this section are hairy, with C. incarnata being the only species with pseudorhiza. Species with two-spored basidia were present on each of the smaller evolutionary branches. Although the spores also had other shapes, such as hexagonal, they were primarily ellipsoid to oblong in shape. Distinct germ pores have also been used to classify certain species. Whether the caulocystidia are lecythiform is the most important feature used to distinguish between different sections of the Conocybe [Conocybe, this represents an evolution from the sect. Pilosellae to sect. Conocybe, with the sect. Mixtae forming the transitional section [Pilosellae were frequently confused with species in the sect. Mixtae, which is why some species in the sect. Pilosellae have been categorized in the sect. Mixtae [Pilosellae into different series.The Bayesian phylogenetic tree, based on the combined ITS, nrLSU, and tef1-α sequences  of the kConocybe . At the  section . The fin. Mixtae ,58. Pile. Mixtae . The evoPilosellae . Within the clades C. muscicola, Conocybe sp.4, and C. incarnata, C. muscicola grows on moss layers, Conocybe sp.4 grows on cow dung, and C. incarnata grows on the ground in broad-leaved forests. Despite their different habitats, they all had reddish-hued pilei. Although the pilei of many species are hygrophanous, meaning that their color changes with the degree of moisture, the importance of color does not decrease. Further research is needed to determine whether this evolutionary branch can be established as a series. The pilei of C. ceracea and C. velutinomarginata have similar features; however, the waxiness of the C. ceracea pileus is distinct, thus being easily distinguishable from C. velutinomarginata [C. reniformis is the presence of reniform spores, resulting in the formation of a separate evolutionary branch. Additionally, C. pilosa formed the sister group of Conocybe sp.2; however, it can be easily distinguished from the latter by its densely hairy pileus and stipe and smooth lamellar edges. The most distinctive feature of C. pseudocrispa is its white pileus with no pigment at the base of the pileipellis. However, the presence of pseudoparaphyses suggests that this is not the only sectional characteristic of the sect. Candidae; for instance, deliquescence is another sectional characteristic [Further phylogenetic analysis of the new species of the sect. losellae  revealedt colors . C. sinoarginata . A typicteristic ,57.Pilosellae from China and their corresponding sequences. This study attempted to integrate morphological classification with molecular systematics, but did not conduct a systematic study on Conocybe Clade1. Therefore, the sect. Pilosellae was not revised, and only some key information is provided, laying the foundation for future research. Currently, some species in the sect. Pilosellae have a few lecythiform caulocystidia at the top of the stipe, which can easily be misidentified as those in the sect. Mixtae. Meanwhile, some species possessing the characteristics of the sect. Pilosellae have been found in other sections [Conocybe species that integrates morphological and systematic approaches to elucidate the relationships between different sections is necessary.This article provides a comprehensive list of all the currently known species in the sect. sections ,58. To a


Here, we present a deeper study of these sequences in Vaccinium L. Molecular-genetic and bioinformatics methods were applied for sequencing, assembly, and analysis of the rolB/C-like gene. The rolB/C-like gene was discovered in 26 new Vaccinium species and Agapetes serpens (Wight) Sleumer. Most samples were found to contain full-size genes. It allowed us to develop approaches for the phasing of cT-DNA alleles and reconstruct a Vaccinium phylogenetic relationship. Intra- and interspecific polymorphism found in cT-DNA makes it possible to use it for phylogenetic and phylogeographic studies of the Vaccinium genus.A variety of plant species found in nature contain agrobacterial T-DNAs in their genomes which they transmit in a series of sexual generations. Such T-DNAs are called cellular T-DNAs (cT-DNAs). cT-DNAs have been discovered in dozens of plant genera, and are suggested to be used in phylogenetic studies, since they are well-defined and unrelated to other plant sequences. Their integration into a particular chromosomal site indicates a founder event and a clear start of a new clade. cT-DNA inserts do not disseminate in the genome after insertion. They can be large and old enough to generate a range of variants, thereby allowing the construction of detailed trees. Unusual cT-DNAs (containing the  Agrobacterium” is a soil bacterium with a unique mechanism to transfer well-defined DNA fragments (T-DNAs) into the chromosomes of a large variety of dicotyledonous plant species [Agrobacterium tumefaciens Conn or A. vitis Ophel and Kerr) or hairy roots formation . It has been experimentally demonstrated that hairy roots can regenerate into fertile plants [Nicotiana glauca Graham and other Nicotiana L. species [Linaria Mill. [Ipomoea L. [Cuscuta L. [Linaria [Ipomoea [Nicotiana [Camellia [Camellia [“ species ,3,4,5,6.e plants ,8,9. Mor species ,11,12, aia Mill. , Ipomoeaomoea L. , Cuscutascuta L. , and dozscuta L. ,17,18. Tscuta L. . Sequenc[Linaria , Ipomoea[Ipomoea , Nicotiaicotiana , and CamCamellia . They prCamellia . PhasingCamellia . These aCamellia .rolB/C-like gene in the genome data of Vaccinium corymbosum L. and Vaccinium macrocarpon Ait. [plast-gene family. It is more similar to plast-genes present in fungus Laccaria bicolor (Maire) P.D.Orton (and absent in most basidiomycetes), plant Nyssa sinensis Oliv., and bacteria Ensifer sp. Casida, than to ones from known strains of Agrobacterium (Rhizobium) rhizogenes, which indicates the transfer of this sequence from an unknown «Agrobacterium» strain [We also found unusual sequences of the pon Ait. ,16. ThisVaccinium genus, using the rolB/C-like gene sequence. This task is quite relevant since the phylogeny of Vaccinium genus is highly controversial.Based on the results of previous phylogenetic studies, we can try to reconstruct phylogenetic relationships between species of Vaccinium genus was first described by Carl Linnaeus in 1753 [Vaccinium phylogeny and taxonomy mainly involved species distributed in North America. The main difficulties in such studies were caused by the absence of fertility barriers in morphologically different organisms, which leads to the formation of a large number of hybrids, along with polyploidy distribution throughout the genus [The  in 1753 . After t in 1753 ,27. TherVaccinium genus in 1945 [Cyanococcus included 9 diploid, 12 tetraploid, and 3 hexaploid species [V. corymbosum with three levels of ploidy; therefore, in the new Kloet’s classification, the Cyanococcus section contained 6 diploid, 5 tetraploid, and 1 hexaploid blueberry species [Camp was one of the first to introduce the system of  in 1945 . He cond species . Later,  species , while d species .Vaccinium species differentiation is complicated by polyploidy, similar morphology, and introgressions during hybridization [Thereby, dization . AdditioVaccinium genus. They allowed us to reconsider the taxonomy proposed by K. Kloet. An example of such study is the application of matK and ITS markers to determine phylogenetic relationships of various representatives of the entire tribe Vaccinieae [Vaccinium and Agapetes); Bracteata-Oarianthe (represented by the respective sections); Orthaea/Notopora (containing the genera Orthaea Klotzsch and Notopora Hook); Myrtillus; and Vaccinium clades. Moreover, most of the discovered clades combined representatives of various genera, while the clades Vaccinium and Myrtillus contained species from different Vaccinium sections according to the previous classifications. Based on the obtained results, the authors concluded that it is necessary to reassess the taxonomy of the genus Vaccinium, since it is not monophyletic. Although work on this reassessment began in 2003 [matK, in taxon, where hybridization and polyploidization played a significant evolutionary role [Methods such as DNA barcoding, fingerprinting, phylogenomics, and chemosystematics have been used recently in the study of ccinieae . The res in 2003 , a numbeary role .Vaccinium genus, more time-consuming and expensive methods of molecular phylogenetics were used in genetic studies of economically important species. Recent assemblies of the genomes of V. macrocarpon, V. microcarpum (Turcz. ex Rupr.) Schmalh., V. oxycoccos L., and V. corymbosum made it possible to conduct their comparative genomics [V. macrocarpon diverged from V. oxycoccos 2 mya, and from V. microcarpum 4.5 mya. Divergence time estimated for V. macrocarpon and V. corymbosum differs from 5 to 10.4 mya [Since the use of DNA barcoding does not enable unambiguous reconstruction of the phylogeny of genomics ,35. Mole10.4 mya ,35.V. darrowii Camp, V. elliottii Chapm, V. tenellum Ait., V. angustifolium Ait., V. corymbosum, V. constablaei A. Gray, V. virgatum Ait, V. myrtilloides Michx., and V. pallidum Ait. The trends identified in this way are consistent with the history of blueberry breeding [An analysis of the intra- and interspecific variability of American blueberry species by double digest restriction-site-associated DNA sequencing , showed breeding .Vaccinium by various methods have led to contradictory results. The use of phylogenetic markers ITS and matK leads to the construction of a phylogenetic tree with significant differences from the classical system. The use of more expensive and time-consuming methods of genome studies gives a more plausible picture, that is more consistent with the classical system. However, expensive methods can currently be applied to a narrow list of economically important species. For more extensive studies of various representatives of the genus, easy-to-use and cheap markers are required. They can be developed based on genome regions newly acquired in the course of evolution with a known structure and localization. An example of such sequences would be cellular T-DNA. Therefore, our study aims to characterize cellular T-DNA polymorphism in the genomes of plants of Vaccinium genus, and to evaluate the possibility of its application for phylogenetic studies of the genus.Thereby, attempts to reconstruct the phylogenetic relationships of species of the genus rolB/C-like gene has been described only in V. corymbosum and V. macrocarpon. In the framework of this study, using BLAST algorithm, this gene was described in the WGS data for V. myrtillus and V. darrowii. Their ORFs correspond to the amino acid sequences from 290 (in V. myrtillus) to 292 amino acids (in V. macrocarpon).Until recently, the Vaccinium genus.In blueberries, the gene is located on chromosome 3 . Located on both sides of it, at a distance of 5 kb, are genes of plant origin with functions that have not been described yet. No other agrobacterial genes were found in the genome of plants of rolB/C-like gene in SRA database revealed new species of naturally transgenic plants in the Vaccinium genus, summarized in Our search for homologues of the Oxycoccoides, Oxycoccus, Cyanococcus, Hemimyrtillus, Myrtillus, Polycodium, Bracteata, Vaccinium, and Vitis-idaea, indicating that their common ancestor was transformed before the sections diverged.Natural GMOs were found in the sections V. selerianum, V. shastense, V. striicaule, V. symplocifolium, V. vidalii, and V. whitmeei), the transgene was not found at all, but this does not mean its absence from the genome, it indicates, however, that additional studies are required.Genomic data for different species varies in coverage quality. In some cases, the presence of a transgene can be inferred from single reads. In other cases , only full-length sequences of the roB/C-like gene were found.Sequences with good coverage were used further to assemble full-length sequences. For a deeper understanding of the gene variability, field material was collected, which was included in further variability analysis of the Vaccinium. They are represented by substitutions and indels. Moreover, indels are equal or multiples of three nucleotides. This is evidence in favor of the gene product functionality. Examples of such polymorphism can be SNPs in V. uliginosum and V. vitis-idaea shown in Vaccinium species shown in When studying SNPs within full-length sequences, one can see that there are nucleotide differences characteristic of the species, distinguishing each of them from other representatives of the genus Vaccinium, as well as in Laccaria, Nyssa, and Ensifer. The closest of the described agrobacterial proteins is the RolB protein  are homogeneous, despite the wide geography of sampling. At the same time, clades D and H are mosaic. Clade H is represented by alleles from involved . Clade GV. macrocarpon and V. oxycoccos are related species that form clade D. These species require more detailed study in the future, since the American cranberry is a cultivated species, and, therefore, hybridization was used in the early stages of its breeding. In turn, swamp cranberries are the only species known today where the transgene was extensively mutated in many of the studied samples.In general, our studies have shown that, at least in the analyzed species, the intraspecific variability of transgene is lower than interspecific, and the mosaic distribution of alleles on the tree in American blueberries can be explained in terms of interspecific hybridization documented during the creation of studied varieties.Therefore, we compiled consensus sequences, as a characteristic of each of the species analyzed at the previous stage, and supplemented this list with sequences obtained on the basis of gene sequences we obtained from single samples of the collection of the Komarov Botanical Institute.rolB/C-like gene of species studied in the previous stage. Alleles of presumably hybrid origin were excluded at the stage of constructing consensus sequences.In the final part of this study, several types of data were used. First of which, were the consensus sequences of the In addition, sequences of species sufficient for gene assembly but not sufficient for allele phasing were included in our analysis.rolB/C-like gene from additional species, represented by single samples in the collection of the Komarov Botanical Institute.Finally, we obtained sequences of the V. myrtilloides (Sec. Cyanococcus), V. ovalifolium Sm. (Sec. Myrtillus), V. praestans Lamb. (Sec. Praestantia), V. smallii A. Gray (Sec. Hemimyrtillus), V. vulcanorum Kom. (Sec. Vaccinium), V. conchophyllum Rehder, V. emarginatum Hayata (Sec. Conchophyllum), and Agapetes serpens (Wight) Sleumer  [The phylogenetic tree presented in logical) .Vaccinium genus are clustered together when constructing a tree based on the proposed DNA marker. Marked with ovals in As can be seen from V. macrocarpon and V. oxycoccos belong to the section Oxycoccus, but on ITS tree these species are separated by species V. vitis-idaea, which belongs to the section Vitis-idaea, Clade Myrtillus unites species of sections Myrtillus and Macropelma. Clade Bracteata-Oarianthe also includes species from sections Hemimyrtillus, Oarianthe, Bracteata, and Baccula-Nigra. Clade Agapetes includes representatives of genus Agapetes and some representatives of genus Vaccinium from sections Rigiolepis, Conchophyllum, and Galeopetalum, while some other species from these sections are spread over the tree. Orthaea/Notopora clade includes representatives of different genera, Notopora, Orthaea, and Vaccinium. The combination of Vaccinium representatives with other genera is also characteristic of Meso-American/Caribbean clades, which includes a representative of section Oreades.Another phylogenetic tree was constructed based on ITS sequences of samples studied in this work, combined with sequences from Kron et al. research . This trrolB/C-like gene.There are much fewer of such contradictions in the phylogenetic tree based on the Vaccinium is the fifth genus of natural GMOs after Nicotiana, Linaria, Ipomoea, and Camellia, where the cT-DNA structure was characterized in greater detail in many species. [species. ,13,14,23plast-gene found in plant genomes, a distinguishing feature of the Vaccinium genus is also the fact that the gene has been preserved intact in most of the studied species. The plast-genes family includes most of the oncogenes of R. rhizogenes , and some T-DNA genes of A. tumefaciens  [Plast genes were described in detail in the review by Leon Otten [plast genes could have similar functions because of their common ancestry, and their diversification could be an adaptation to different plant species. Describing the biodiversity of plast genes can be very important in order to find their basic function and some common features of all these genes. In this regard, the discovery of plast genes, that are not quite similar to previously known ones (as in our case), is very valuable for future research.In addition to the unusual  6a, 6b) . They ha 6a, 6b) ,13,14,38on Otten  suggesteVaccinium subgenera and Agapetes have a common localization site of the rolB/C-like sequence in the genome. This indicates a common origin of the sequence, as a result of a single transformation event of the ancestral form, and, accordingly, the monophyletic origin of all species containing transgene, included in our analysis. During evolution, the newly acquired sequence seems to have accumulated mutations. This variability can be assessed and used to estimate species divergence.In accordance with the analysis of previously assembled genomes and sequences, the representatives of two Vaccinium genus since there is still no consensus on the genus system.This is very important for the Vaccinium genus have been conducted since the middle of last century [matK, revealed contradictions with the traditional system. The trees constructed using ITS sequences by Kron et al. [Vaccinium and Agapetes; Bracteata-Oarianthe, uniting representatives of the respective sections; Orthaea/Notopora, which includes the genera Orthaea and Notopora; Myrtillus; and Vaccinium. These clades contained species of the genus Vaccinium, which, in previous classifications, belonged to different sections. Moreover, representatives of the oxycoccus subgenus were separated on the tree by V. vitis-idea. Representatives of the Cyanococcus section also fell into different clades, clustering with species of other sections.Phylogenetic studies of the  century ,27. The  century . The usen et al. , and recVaccinium.This contradiction could be related to the manifestation peculiarities of used DNA markers in species of hybrid origin. The ITS of hybrid species retain the features of only one parent, and chloroplast markers are inherited mainly through the maternal line . At the Camellia genus [Vaccinium plants, as well as to identify homo- and heterozygotes for this marker. This may be useful in interpreting controversial results of the origin of hybrid forms, as well as illustrating the incomplete isolation of species, as previously demonstrated in case of the genus Camellia [In this article, we propose such a marker. An important feature of the work carried out by us was the use of a large number of forms of cranberries, lingonberries, and blueberries of different geographical origin, as well as the phasing of natural transgene alleles using SRA data in order to assess intraspecific variability. Such an approach was first used to study cellular T-DNA of representatives of the ia genus . The anaCamellia .Vaccinium is less than the interspecific one. Therefore, it can be used to study the phylogenetic relationships of Vaccinium species.Based on the phylogenetic analysis of allelic variants, obtained in our research, it can be concluded that intraspecific variability of marker in Vaccinium was performed, using sequences of the rolB/C-like gene, as well as ITS sequences from previous studies [A phylogenetic analysis of the genus  studies , supplemrolB/C-like gene shows that they have similarities and differences. Species of Myrtillus section  cluster together in both trees. Species of Cyanococcus section are clustered on both trees in a similar way. No other similarities in the topology of these trees were found. Species V. conchophyllum and V. emarginatum belonging to section Conchophyllum cluster together on the rolB/C-like tree, but not on ITS tree. Species V. macrocarpon and V. oxycoccos from section Oxycoccus, V. vulcanorum and V. uliginosum from section Vaccinium form separate clades on the rolB/C-like tree, but are scattered across different parts of the ITS tree.The comparison of trees built based on ITS and Bracteata, Hemimyrtillus, Vitis-idaea, Oxycoccoides, and Praestantia sections are represented by single species on the rolB/C-tree. These sections require additional research involving new species. Reconstruction of phylogeny using the rolB/C-like gene is consistent with data obtained by other researchers using more expensive and time-consuming methods [ methods ,35,42.rolB/C-like gene, has fewer contradictions with classical ideas, compared to a tree built based on ITS. The rolB/C-like gene, as a molecular marker, is cheap, easy to use, and is not time consuming. Given minimal amounts of biological material, new species can easily be included in further analyses.Therefore, a phylogeny, built on the basis of the Vaccinium myrtillus L., V. oxycoccos, V. vitis-idaea L., and V. uliginosum L., collected from various habitats in Russia and Belgium. Samples of 9 species of the studied genus were taken from the collection of the Komarov Botanical Institute . A detailed description of the samples is presented in Plant material was represented by samples of Vaccinium species from SRA database, used in this study are presented in Sequences of Leaves and stems of plants were grounded in liquid nitrogen. Plant DNA was isolated by a CTAB method described by Draper and co-authors .Type I. Classic PCR was performed using the primers shown in E. coli bacterial cells taken from a colony of transformants. The following program was used for real-time PCR: 60 s at 50 °C, 40 cycles of 5 min at 95 °C, 30 s at 58 °C, 60 s 72 °C, and 18 s at 95 °C.Type II. Real-time PCR from colonies was performed for additional control of early threshold values to search for clones containing the target insert. Real-time PCR was carried out in ANK-32 cycler . Primers for plasmid pJET1.2 from the CloneJET PCR Cloning Kit  were used in the reaction. The 20 μL of the reaction mixture contained SsoAdvanced Universal SYBR Green Supermix buffer , 10 μM primers, and a suspension of PCR products were separated on agarose gel in 1x TBE buffer and visualized using GelDoc Go .PCR products were cloned into pJET1.2 vector, using CloneJET PCR Cloning Kit  according to the manufacturer instructions, and transformed into DH5 alpha chemically competent cells according to Inoue and co-authors .PCR fragments were sequenced using Sanger method, and BrilliantDye™ Terminator (v3.1) Cycle Sequencing Kit . Then, sequencing mixtures were separated at the Resource Center of St. Petersburg University “Development of Molecular and Cellular Technologies” using an ABI Prism 3500 xl sequencer .In order to determine the sequences of alleles, we represented in binary form each sample as a set of polymorphic positions, where “1” indicates the most frequent nucleotide in a given position in the species, and “0” the least frequent. Then, for each sample, all possible combinations of values were written for polymorphic positions. Since homozygotes and samples with one SNP were found among the samples, we were able to obtain a primary pool of alleles, and these alleles were subsequently found in the remaining samples. Since each of these alleles, in a diploid, must match to an allele with alternative values in polymorphic positions, a homologous pair can be found for the primary allele .In cases where the number of sequences was insufficient to resolve the set of sequences per allele, they were cloned into pJET 1.2 according to Clone Jet PCR Cloning Kit  manufacturer’s instructions. Then, inserts from individual clones, that represent individual alleles, were sequenced.Vaccinium genus. The search was carried out using BLAST against the reference (JOTO01169953.1). Resulting reads were aligned to the reference using BWA 0.7.17 [The NCBI SRA database was used to search for T-DNA in the A 0.7.17 . The proA 0.7.17 . The aliA 0.7.17 . Allele A 0.7.17  followedA 0.7.17 . AdditioSequences were aligned using the MAFFT online service for multiple sequence alignment . The evohttps://colab.research.google.com/github/sokrypton/ColabFold/blob/main/AlphaFold2.ipynb (accessed on 18 April 2022), using default settings.AlphaFold prediction  of the sVaccinium and Agapetes. The natural transgene of these plants can be used for phylogenetic studies of the genus. This nuclear marker is cheap, easy to use, and is not time consuming. It can also be recommended for further research. The allele phasing approach makes it possible to track hybridization events in the evolution of studied plants. Data, regarding intraspecific variability of the proposed marker, can be used to mark populations of wild berries of the Vaccinium genus, in order to prevent their unauthorized harvesting.In this study, new species of natural GMOs were described within the genera


Murugan  RSC Advances article due to concerns with the reliability of the data in the published article.The Royal Society of Chemistry, with the agreement of the named authors, hereby wholly retracts this There are unexpected similarities between the FTIR spectra in Fig. 1a, b and d, which represent different samples.The HRSEM image in Fig. 3c represents a scaled version of part of the image presented in Fig. 3b. However, both images are presented with the same scale bar and represent different experimental conditions.The authors informed the Editor that the characterization of the original samples was outsourced, and they do not have the original raw data for the published results.Given the significance of the concerns about the validity of the data, and the lack of raw data, the findings presented in this paper are not reliable.N. Murugan and K. Vimala were contacted but did not respond. S. Kannan responded but did not confirm whether they agreed to retract the article.Signed: L. Kavitha, E. Shinyjoy, D. Rajeswari and D. GopiDate: 16th March 2023RSC AdvancesRetraction endorsed by Laura Fisher, Executive Editor,


Ehrlichia chaffeensis is a tick-borne bacterium that causes human monocytotropic ehrlichiosis, an emerging life-threatening disease in humans transmitted by Amblyomma americanum. Although most studies have reported bacterial isolations and clinical cases in the United States, their occurrence is not restricted to North America. Some studies in the Southern Cone of South America have molecularly detected a close phylogenetic relative of E. chaffeensis in ticks and wild mammals. Even so, many gaps must be filled to confirm the presence of this agent in the region. To add new data on this issue, we report the first detection of specific anti-E. chaffeensis antibodies in dogs collected from all regions of Brazil. By means of IFA and ELISA with crude and specific antigens of E. chaffeensis, sera from 1134 dogs were analyzed. Serological analyses using ELISA showed nine (0.7%) seropositive dogs, with seven of them exhibiting IFA titers ranging from 160 to 5120. All regions of Brazil had at least one seropositive dog. Our results support the evidence for the occurrence of E. chaffeensis in South America. As dogs have a close relationship with humans, they can be used as an environmental sentinel for these infections because they can act as a bridge to human parasitism or infection with ehrlichial agents. Ehrlichia chaffeensis is a tick-borne bacterium that causes human monocytotropic ehrlichiosis (HME), an emerging life-threatening disease in humans that typically results in fever-like symptoms but may also be fatal if left untreated. In the United States, this infection has been increasing over the years, mainly in the states of the southeast and Midwest, as well as much of the east coast, where populations of white-tailed deer (Odocoileus virginianus), the main reservoir of E. chaffeensis and host of the lone-star tick (Amblyomma americanum), the vector of this disease, abound [E. chaffeensis [Ehrlichia spp. that are similar or closely related to E. chaffeensis have been detected in ticks, namely, Amblyomma parvum [Amblyomma tigrinum [Rhipicephalus microplus, as well as in marsh deer (Blastocerus dichotomus) [Mico melanurus) [Ehrlichia spp., abound . In addiffeensis . For thia parvum , Amblyomtigrinum , and Rhihotomus)  and non-lanurus) . A recenE. chaffeensis can be performed by using antibody research and/or molecular detection [E. chaffeensis to avoid false-positive results. In this sense, the TRP32 protein was previously molecularly characterized as being specific to E. chaffeensis, and anti-TRP32 antibodies did not cross-react with the closely related E. canis or other Ehrlichia species [E. chaffeensis and has been employed in serological surveys in the USA, Canada, and the Caribbean [The diagnosis of canine infection by etection . The lat species . Therefoaribbean .E. chaffeensis are scarce. As canines have a close relationship with humans, they are susceptible to different species of ticks and E. chaffeensis; therefore, they can be considered a bridge to human parasitism by ticks. Given the relevance of this issue, in this communication, we present the first serological evidence of E. chaffeensis in dogs from Brazil.In Brazil, although serological evidence of human infection by this agent has been reported , studiesE. chaffeensis antigen with an immunofluorescence assay (IFA) [E. chaffeensis-specific antigen (TRP32) with an enzyme-linked immunosorbent assay (ELISA) [With reference to publications by Melo et al. ,12 and T (ELISA) . This stE. chaffeensis with a cut-off point at an initial dilution of 1:64 [An immunofluorescence assay (IFA) was used to detect antibodies on slides that were sensibilized with DH82 cells infected with the Arkansas strain of  of 1:64  that werE. chaffeensis  [E. canis  [E. chaffeensis infection in IFA had mean OD readings of <0.050; therefore, a positive sample threshold was set at >0.150 OD units above the negative control absorbance.Peptides corresponding to the immunodominant repeat regions of TRP32 from EAVQLGN)  were synLYTQLTL)  was usedLYTQLTL) . The devE. chaffeensis antibodies were evaluated by using the Chi-square test or exact Fisher test, and p ≤ 0.05 was considered significant. The statistical analysis was performed using Epi InfoTM version 5.5.1.The differences between the results found in each region and positivity for E. chaffeensis in all Brazilian regions, so there were no statistically significant differences (p > 0.05). The identification, antibody titers, and OD values for each sample and region are shown in The serological analyses showed that there were nine (0.7%) seropositive dogs according to the ELISA assay, and seven (0.6%) of them had IFA antibody titers ranging from 160 to 5120. All regions of Brazil had at least one reactive dog . SimilarE. chaffeensis: DH82 cells infected with an Arkansas isolate were used for an indirect immunofluorescence reaction [E. chaffeensis may share similar epitopes with E. canis or Ehrlichia minasensis, which are other members of the Ehrlichia genus that are endemic among dogs (E. canis) [E. minasensis) [E. chaffeensis are characterized by their high specificity; moreover, the TRP32-ELISA also guaranteed an appropriate sensitivity when detecting samples in the context of seroconversion, even in comparison with the IFA technique, as can be seen in two of the samples shown in E. canis [In this survey, we used two different types of antigens to assess the presence of antibodies against . canis)  or preseasensis)  in BraziE. canis .E. chaffeensis infection but could also come from infection by a closely related species. The South American isolates should be more extensively investigated, especially for other genetic targets, as the detections in most studies have been based on one or two genes  [E. chaffeensis or a closely related species in South America, and this is the first survey to evaluate the occurrence of this pathogen in dogs in Brazil through the detection of antibodies.Although the specificity of the TRP32 peptide was supported by a previous experiment , we prefNA, dsb) ,5,6. HenE. canis antibodies detected in dogs in Brazil, with rates ranging from 16 to 57% [E. chaffeensis demonstrated a low occurrence of this agent among dogs. Beall et al. [E. chaffeensis antibodies ranged from 0.1 to 21.4% depending on the location evaluated and the presence of A. americanum.The samples that were tested came from all states of Brazil except for the state of Amazonas; i.e., the survey was comprehensive and involved all Brazilian ecosystems and biomes. In contrast to the findings on the prevalence of anti-6 to 57% , the frel et al.  evaluateB. dichotomus), one of the probable hosts of this species of Ehrlichia, has been reported [The presence of seropositive dogs from all regions of Brazil also suggests that the occurrence of this species is not restricted to the Pantanal, other flooded Brazilian regions, or even the Southern Cone of South America , as these are the regions in which the presence of marsh deer  [E. chaffeensis antibodies in dogs from regions in which this species does not occur would not be justified. Rhipicephalus microplus is predominantly distributed in tropical and subtropical areas worldwide and is strongly associated with cattle. Nevertheless, there are records of R. microplus in various hosts, from other mammals and birds to amphibians [E. chaffeensis was found in specimens that parasitized B. dichotomus [Although our results reinforce the hypothesis of the presence of . parvum , A. tigrtigrinum , and R. icroplus  ticks fral areas , and theal areas ,10,11,12America) . Therefophibians . Howeverchotomus .E. chaffeensis was previously detected in Rhipicephalus sanguineus sensu lato ticks in Cameroon, where A. americanum is not found [E. chaffeensis to dogs by R. sanguineus s. l. was experimentally observed [R. sanguineus s. l. in Brazil, this tick must be intensely investigated for other species of Ehrlichia in addition to E. canis.Interestingly, ot found ,21. Addiobserved . TherefoE. chaffeensis or a closely related species. Due to the different characteristics of the states and their regions, the probable transmitting ticks of these species of Ehrlichia must be further investigated. The data presented in this communication reinforces the need to discuss tick control strategies, as these parasites, which are deeply involved in animal health, can transmit important pathogens to humans. Due to the disorders caused by E. chaffeensis in the United States of America and developing countries, such as Brazil, which has several areas with sanitary and environmental deficits, infections transmitted by ticks can have negative impacts on public health.We conclude that dogs from different areas of Brazil were exposed to


Correction: BMC Sports Sci Med Rehabil 15, 54 (2023)10.1186/s13102-023-00667-7Following publication of the original article , the autMCH, NRH, JMH, DMT, PHS, AS, and JJF report grants from Congressionally Directed Medical Research Programs and Office of Naval Research, outside of the submitted work. PAG, KBK, KLT report grants from Congressionally Directed Medical Research Programs, outside of the submitted work. NRH, PAG, and JMH report funding from the Federal Emergency Management Agency, outside of the submitted work. KLT reports grants from the National Institutes of Health and the National Science Foundation, outside of the submitted work. DL reports grants from the National Institutes of Health, outside of the submitted work. In addition, PHS, AS, and JJF have a patent pending for an Adaptive and Variable Stiffness Ankle Brace, U.S. Provisional Patent Application No. 63254,474.PHS, AS, and JJF are military service members or employees of the U.S. Government and this work was prepared as part of their official duties. Title 17, U.S.C. § 105 provides that copyright protection under this title is not available for any work of the U.S. Government. Title 17, U.S.C. § 101 defines a U.S. Government work as work prepared by a military service member or employee of the U.S. Government as part of that person’s official duties. The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, nor the U.S. Government. The study protocol was approved by the University of Kentucky Institutional Review Board in compliance with all applicable Federal regulations governing the protection of human subjects, number 58,500.


Menispermi Rhizoma, the rhizome of Menispermum dauricum DC., is a traditional Chinese medicine, which has the effect of clearing away heat and detoxification, dispelling wind, and relieving pain. It is often used in the treatment of sore throat, enteritis, dysentery, and rheumatism. The chemical constituents of M. Rhizoma mainly include alkaloids, phenolic acids, quinones, cardiotonic glycosides, and so on. Modern pharmacological studies have proved that M. Rhizoma has the effects of anti-tumour, anti-inflammation, anti-oxidation, bacteriostasis, cardio-cerebrovascular protection, anti-depression and anti-Alzheimer’s disease. In recent years, the chemical constituents of M. Rhizoma have been found continuously, and the pharmacological studies have deepened gradually. This paper reviews the research progress on the chemical composition and pharmacological effects of M. Rhizoma, to provide a basis for further research and development of its medicinal value. Menispermi Rhizoma is the dried rhizome of Menispermum dauricum DC. It is mainly produced in Northeast China, North China, East China, and Shaanxi. It has the effect of clearing heat and detoxifying, dispelling wind, and relieving pain, and is mainly used for sore throat, pyretic diarrhoea, dysentery, and rheumatic paralysis -quinoline, oxidized isoapocynine alkaloids oxoisoaporphine B, menisoxoisoaporphine A, and daurioxoisoporphine B in M. Rhizoma showed good inhibitory activity against the release of NO from rat macrophages in the LPS-induced anti-inflammatory activity assay [M. Rhizoma inhibit ovalbumin-induced airway inflammation in mice with asthma by reducing the concentrations of interleukin 4, 5, and 13, down-regulating the levels of TNF-α and eotax in bronchoalveolar lavage fluid, and inhibiting the increase in serum levels of total immunoglobulin E and ovalbumin-specific immunoglobulin E. The results of this experiment suggest that the total alkaloids of M. Rhizoma can inhibit ovalbumin-induced airway inflammation in mice by modulating T-helper 2 responses and chemokine levels, suggesting that the total alkaloids of M. Rhizoma may be potential anti-asthmatic agents [M. Rhizoma, especially bisbenzylisoquinoline and morphinane alkaloids, are the significant pharmacological bases for the anti-inflammatory activity of M. Rhizoma. In addition, arachidic acid obtained from the methanolic extract of M. Rhizoma by Ren et al. [M. Rhizoma.Ulcerative colitis (UC)—characterized by abdominal pain; diarrhoea; and mucous, bloody stools as the main clinical manifestations—has a high recurrence rate and is difficult to cure . Studieshe colon . Similarhe colon . Networkholidine . In addity assay ,17. In ac agents . In summn et al.  showed aM. Rhizoma [M. Rhizoma also had a better scavenging ability for DPPH radicals with a scavenging rate of 70.1%; thus, it was speculated that the long-chain unsaturated fatty acid methyl esters such as methyllinoleate and methyloleate, which were more abundant in the fatty oil of M. Rhizoma identified by GC-MS, might be related to the antioxidant activity of the fatty oil of M. Rhizoma.DPPH radicals are commonly used for in vitro antioxidant activity evaluation, and the stronger the scavenging ability of DPPH radicals, the stronger the antioxidant capacity. The scavenging rate of oxidized apomorphine alkaloids dauriporphine and menisporphine on DPPH radicals was similar to that of the positive reference drug vitamin C, both above 90%, providing an experimental basis for the development of antioxidant drugs from the alkaloids of  Rhizoma . In addi Rhizoma  found thM. Rhizoma were reported to have inhibitory effects on a variety of respiratory and intestinal bacteria, with the most significant inhibitory effect on dauricine, with an inhibition rate of 83.33%, and the best inhibitory effect on S. pneumoniae [E. coli, S. aureus, and B. subtilis to different degrees, and the inhibitory effect was: B. subtilis > S. aureus > E. coli [The alkaloid components of eumoniae . Clinica E. coli .2+ channels, and its antiarrhythmic mechanism of action is similar to that of the class III antiarrhythmic drug amiodarone: it mildly inhibits Ca2+-ATPase activity, decreases sarcoplasmic reticulum calcium uptake and has the effect of inhibiting Na+ inward flow, Ca2+ inward flow, and K+ outward flow, especially blocking K+ outward flow [Dauricine is often used as a clinical treatment for hypertension and cardiac arrhythmias. Its antihypertensive effect was reported to be related to the antagonism of Caard flow . Ischemiard flow . Zhang eard flow  found thard flow . Other fard flow .M. Rhizoma that the protective effect of bisbenzylisoquinoline and morphine alkaloids on hypoxia-injured EA.hy926 vascular endothelial cells were more obvious. The more abundant bisbenzylisoquinoline alkaloid daurisoline in M. Rhizoma showed the strongest anti-hypoxic activity, followed by morphine alkaloids acutumine and acutuminine. The above studies provide the material basis for the better anti-hypoxic activity of M. Rhizoma.Shao et al.  found inM. Rhizoma [M. Rhizoma are expected to be developed as more efficient antidepressants.Depressed patients tend to have decreased levels of 5-hydroxytryptamine (5-HT), and certain genetic polymorphisms in 5-HT metabolism and transporters are associated with depression . Studies Rhizoma . The com Rhizoma . Based oM. Rhizoma inhibited acetylcholinesterase (AChE), with the monobenzylisoquinoline alkaloid pecrassipine B having the most significant inhibitory effect on AChE, followed by that of the bisbenzylisoquinoline alkaloid daurisoline, the morpholino alkaloid acutumine, the simpleisoquinoline alkaloid thalifoline, pycnarrhine, and amurolin; the inhibition effect of the simpleisoquinoline alkaloid corypalline is weaker in comparison. The molecular docking results showed that the strength of AChE inhibition by pecrassipine B and corypallinewas was related to their respective molecular structures and the degree of AChE binding [M. Rhizoma: Wang [M. Rhizoma are potentially promising for the prevention and treatment of AD.One of the primary pathogeneses of Alzheimer’s disease (AD) that is now widely recognized is its association with impairment in cholinergic transmission processes, where patients with low acetylcholine levels and reduced function in the brain experience significant cognitive impairment. The results of in vitro enzyme activity experiments showed that the alkaloids in  binding . Neuroto binding . It has  binding . Two othma: Wang  used thema: Wang . In summM. Rhizoma is slightly toxic and clinical application is accompanied by adverse effects such as nausea, vomiting, loss of appetite, dyspepsia, bloating, and diarrhoea. Studies have shown that the acute toxicity of the alcoholic fraction of M. Rhizoma is greater than that of the aqueous fraction [M. Rhizoma are the major alcohol-soluble components, thus verifying the studies on the chemical composition of M. Rhizoma reported in the literature [M. Rhizoma are the main material basis for its toxicity. The toxic effects of the aqueous and alcoholic fractions of M. Rhizoma manifested as acute or chronic hepatotoxic injury with significant changes in serum ALT, AST, and hepatic body ratios [M. Rhizoma can increase MDA content in liver tissue and decrease SOD activity; this confirms, at the intrahepatic substance level, that the mechanism of hepatotoxic injury caused by M. Rhizoma is related to the induction of lipid peroxidation and reduction of its own redox capacity after causing oxidative stress in the body, as well as to the NO-mediated damage pathway [fraction , and theterature . This suy ratios . Similar pathway .M. Rhizoma has a long history of medicinal use. In recent years, domestic and foreign scholars have conducted extensive and in-depth studies on the chemical components of M. Rhizoma, especially alkaloid components, and up to now, more than 150 chemical components, including 117 alkaloids, have been identified from M. Rhizoma. Among the alkaloid components of M. Rhizome, bisbenzylisoquinolines are predominant, and apomorphines and oxidized isoporphines are the next most abundant; the effects of M. Rhizoma in anti-tumour, antioxidant, anti-inflammatory, antibacterial, cardiovascular and cerebrovascular protection, anti-depressant, and anti-Alzheimer’s disease have been gradually confirmed and applied in the treatment of clinical diseases. The above research results have carried forward the modernization of traditional medicines. To summarize the current results, three points need attention for further research on M. Rhizoma: Firstly, in the pharmacological research on the alkaloid components of M. Rhizoma, most of the studies focused on the PAMD and relatively few studies on other alkaloids. Compared with the current research, the pharmacological studies of other chemical components isolated from M. Rhizoma are relatively lacking. Thirdly, the pharmacological study on M. Rhizoma can also be combined with the knowledge of molecular biology, proteomics, metabolomics, and other disciplines to investigate further the targets, mechanisms, and metabolic patterns of its effects. This paper reviews the progress of research on the chemical composition and pharmacological effects of M. Rhizoma in recent years and provides a basis for the further development and utilization of M. Rhizoma.With a wide distribution range and abundant medicinal resources,


DOI: https://doi.org/10.1039/C4RA04484C.Retraction of ‘Investigation on corrosion protection and mechanical performance of minerals substituted hydroxyapatite coating on HELCDEB-treated titanium using pulsed electrodeposition method’ by D. Gopi  RSC Advances article due to concerns with the reliability of the data in the published article.The Royal Society of Chemistry, with the agreement of the named authors, hereby wholly retracts this Repeating fragments can be observed in the cross-sectional SEM images of Fig. 4c, f and i, indicating that the images have been manipulated.1In addition, parts of the cross-sectional SEM image in Fig. 4c have been duplicated in another publication.The authors informed the editor that the characterization of the original samples was outsourced, and they do not have the original raw data for the published results.Given the significance of the concerns about the validity of the data, and the lack of raw data, the findings presented in this paper are not reliable.R. Pramod and Jishnu Dwivedi were contacted but did not respond.Signed: D. Gopi, A. Karthika, D. Rajeswari and L. Kavitha.Date: 16th March 2023.RSC Advances.Retraction endorsed by Laura Fisher, Executive Editor,


Psychiatric comorbidity has a significant impact on the patient’s overall health, with an increased risk of death for those patients with mental-physical comorbidity . This impacts, among other things, the average hospital stay of a patient with psychiatric comorbidity. For example, an American study shows that psychiatric comorbidity was associated with greater inpatient utilization, including the risk of additional hospitalizations, days of stay, and hospitalization charges . Our study aims to confirm these results in patients admitted to a general hospital for any cause and presenting psychiatric comorbidity.To compare the mean length of stay of patients admitted to a general hospital for any cause according to whether they have psychiatric comorbidity or not.We made a descriptive retrospective study through the use of electronic medical records. The drug use history and average day of hospitalization were obtained for all patients admitted to the inpatient service of a general hospital during a 3-year period.The mean length of stay was longer in patients with psychiatric comorbidity  than in patients without psychiatric comorbidity , the difference being statistically significant for the analysis of variance with a small effect size . The assumption of the equality of variances of the two groups is not fulfilled  so Welch’s nonparametric test was applied, whose results do not modify those obtained.Our results are in line with other studies, showing a longer mean length of stay in those patients admitted for any cause and with associated psychiatric comorbidity. This highlights the importance of having an integrated psychiatry service in a general hospital, as Bronson points out, where they find a shorter mean length of stay in units that have integrated, proactive psychiatric care .Bronson, B. D., Alam, A., & Schwartz, J. E. (2019). The Impact of Integrated Psychiatric Care on Hospital Medicine Length of Stay: A Pre-Post Intervention Design With a Simultaneous Usual Care Comparison. Psychosomatics.Sayers, S. L., Hanrahan, N., Kutney, A., Clarke, S. P., Reis, B. F., & Riegel, B. (2007). Psychiatric comorbidity and greater hospitalization risk, longer length of stay, and higher hospitalization costs in older adults with heart failure. Journal of the American Geriatrics Society.Tan, X. W., Lee, E. S., Toh, M., Lum, A., Seah, D., Leong, K. P., Chan, C., Fung, D., & Tor, P. C. (2021). Comparison of mental-physical comorbidity, risk of death and mortality among patients with mental disorders - A retrospective cohort study. Journal of psychiatric research.None Declared


Macaca mulatta) is one of the most investigated nonhuman primate species in biomedical research since its anatomy and physiology resemble those of humans. This manuscript fulfills the researcher’s and veterinarian’s need for detailed anatomical data on the rhesus monkey’s pelvic limb. Several rhesus monkey cadavers were dissected to study the anatomy of the muscular, circulatory and peripheral nerve systems of the pelvic limb in relation to each other. The anatomical structures are textually described and illustrated by means of numerous detailed colored images.The rhesus monkey (Macaca mulatta) is a widely used model in biomedical research because its anatomy and physiology bear many similarities to those of humans. Extensive knowledge of the anatomy of this nonhuman primate species is not only required for the correct interpretation of obtained research data but also valuable for the welfare of captive individuals housed in, e.g., zoos. As anatomical publications on the rhesus monkey are hardly available, outdated and provide only line drawings or black-and-white photographs, the anatomy of the rhesus monkey was readdressed in this study. The various anatomical structures are described in relation to each other topographically per hindlimb region. The hip region, the upper limb, the knee, the lower limb and the foot are described from various perspectives. The structures that are visible in the different layers, from the superficial to the deepest layer, were photographed. Although the anatomy of the hindlimbs of rhesus monkeys and humans are remarkably similar, various subtle dissimilarities have been observed. Consequently, an open-access publication that focuses on the anatomy of the rhesus monkey would be highly valued by both biomedical researchers and veterinarians.The rhesus monkey ( Macaca mulatta) live in the southeastern parts of Asia, but this species can also be found in research facilities worldwide, as it is one of the most studied nonhuman primates  is obvious on the superficial layer A. This mSince the m. gluteus medius is located, for the most part, deep to the m. gluteus superficialis and the m. tensor fasciae latae, it can be better distinguished after these muscles have been retracted B. It canThe m. piriformis and the m. gluteus profundus [m. gluteus minimus] become visible after the transection of the m. gluteus medius C. The m.The origins of the above-mentioned nerves, called the lumbosacral plexus, can be identified after the m. piriformis is transected and the proximal and distal muscle stumps are retracted D. In thiSubsequently, the m. gluteus profundus was transected and its muscle stumps were removed. In addition, the n. ischiadicus was laterally retracted (and fixated over the greater trochanter of the femur), allowing for the study of the remaining deep structures. The tendon of the m. obturatorius internus was lifted by means of tweezers E. After The most superficial muscles that become visible after skinning the medial side of the hip region are the m. sartorius and the m. gracilis A. TogethThe m. sartorius and the m. gracilis are subsequently transected near their insertions and retracted B. From cRetracting both parts of the m. semimembranosus together with the m. semitendinosus allows for visualizing the two muscle bellies of the m. adductor magnus C. In betRetracting the stumps of the m. adductor longus, as well as transecting and retracting both muscle bellies of the m. adductor magnus, allows for identifying the m. pectineus, the m. adductor brevis and the m. quadratus femoris from cranial to caudal D. The shWhen the m. pectineus and the m. adductor brevis are transected and their stumps are retracted, the entire m. obturatorius externus can be distinguished E. This mIn When the hindlimb is skinned, the m. quadriceps femoris and the hamstring musculature are obvious A. The m.When the m. vastus lateralis is maximally withdrawn from the m. biceps femoris, the m. rectus femoris and the m. vastus intermedius can be examined C. The laThe m. rectus femoris and the a., v. and n. femoralis can be better visualized when the m. vastus lateralis is transected in its middle portion and the stumps are proximally and distally retracted D. Like tThe caudolateral approach to the upper hindlimb allows for examining the hamstring musculature, which is situated caudolateral to the femur, and the sciatic nerve. Removing the skin shows that, from cranial to caudal, the hamstrings consist of the m. biceps femoris, the m. semitendinosus and the m. semimembranosus A. Since The transection of the m. biceps femoris and m. semitendinosus permits the study of the m. semimembranosus B. This mWhen the epineurium that surrounds the sciatic nerve is longitudinally incised, the n. fibularis communis and the n. tibialis appear as two separate nerves that arise from the n. ischiadicus close to the extension angle of the hip C. The n.The medial side of the hindlimb is characterized by the presence of the cranial m. sartorius and the caudal m. gracilis that delineate the trigonum femorale, in which, from cranial to caudal, the n, a. and v. femoralis are situated A. The a.The hamstring musculature can be examined in detail after the m. sartorius and the m. gracilis are transected and their stumps are retracted B. The m.The adductor muscles are studied after the m. semimembranosus and the m. semitendinosus are transected and their stumps are retracted C. The m.The knee was extensively examined by a caudal approach since several blood vessels and nerves pass through the popliteal region. Lateral, medial and caudomedial will be presented. The knee joint is not included in this topographical description, as its anatomy can be consulted elsewhere .The musculature that is visible directly after skinning the caudal side of the popliteal region consists of the proximally located hamstrings and the gastrocnemius muscle, which forms the superficial muscle mass of the lower leg A. The m.When the semitendinosus, semimembranosus and adductor muscles are removed, the blood vessel ramifications on the caudomedial side of the popliteal cavity can be examined B. The v.The lateral retraction of the m. biceps femoris shows that the n. cutaneus surae lateralis laterally branches off the n. fibularis communis. This cutaneous branch can be biopsied for research purposes. The cutaneous nerve that laterally branches off the n. tibialis is the n. cutaneus surae caudalis. It distally joins the v. saphena lateralis in the groove between both gastrocnemius heads. When these heads are separated, the a. poplitea can be examined in the profundity of the popliteal region. It gives off the a. tibialis cranialis [a. tibialis anterior] and continues as the a. tibialis caudalis [a. tibialis posterior], which joins the n. tibialis. The fine veins that run bilateral to the a. tibialis caudalis and present several anastomoses are the vv. comitantes cum a. tibialis caudalis. Now, the lateral m. soleus and the medial m. flexor digitorum (longus) medialis/tibialis [m. flexor digitorum longus] are also exposed. The m. soleus arises from the head of the fibula. Its tendon fuses with the tendon of the lateral head of the gastrocnemius muscle. The m. flexor digitorum medialis/tibialis, which can be considered the m. flexor digitorum superficialis in domestic mammals, arises halfway from the caudal side of the tibia. Its tendon crosses the medial malleolus and splits to attach to the plantar sides of the distal phalanges of digits II to V.The deepest layer, which is obtained by the resection of the m. gastrocnemius caput laterale and caput mediale as well as the m. soleus, presents the m. flexor digitorum (longus) lateralis/fibularis [m. flexor hallucis longus] next to the m. flexor digitorum medialis/tibialis. This muscle can be considered the m. flexor digitorum profundus in domestic mammals, as it is located deeper to the latter. It arises from the caudomedial aspect of the fibula, the interosseous membrane between the tibia and fibula, and the distal part of the tibia. Its tendon runs along the plantar side of the tarsal joint and then splits into three tendons, one each for digits I, III and IV. Deep to the m. soleus, lateral to the m. flexor digitorum medialis/tibialis, the m. tibialis caudalis can be observed. This muscle arises from the caudal side of the tibia. Its tendon crosses the medial malleolus and inserts into the plantar sides of the metatarsal bones of digits II to IV.When the skin of the hindlimb is removed, the following muscles can be recognized from cranial to caudal: m. tibialis cranialis [m. tibialis anterior], m. extensor digiti primi  longus, m. extensor digitorum longus, m. fibularis longus and finally the m. gastrocnemius caput laterale A. The m.The m. biceps femoris and the deep fascia that envelopes the musculature were subsequently removed B. As a rThe deep musculature on the caudal side of the lower limb can be studied after the lateral and medial heads of the gastrocnemius muscle are transected and retracted C. The m.In When the m. tibialis cranialis is pulled cranially and the m. fibularis longus is pulled caudally, the m. extensor digiti primi  longus and the m. extensor digitorum longus become fully exposed. In addition, the distal part of the m. fibularis brevis emerges E.However, the m. fibularis brevis can be described in more detail after the m. extensor digitorum longus is retracted cranially and the m. fibularis longus is pulled caudally F. It ariLike the m. fibularis brevis, the very thin m. fibularis digiti quinti [m. fibularis tertius] arises from the lower two-thirds of the shaft of the fibula. In contrast, it inserts into the metatarsal bone of the fifth digit. Due to its deeper position, it can be seen caudal to the fibularis brevis muscle when this muscle is slightly pulled cranially G.The m. fibularis longus and the m. plantaris are transected in Finally, the sulcus extensorius of the tibia was exposed after the m. tibialis cranialis was transected and the stumps were retracted I. The prThe muscles that can be identified directly after skinning the medial side of the hindlimb are, from cranial to caudal, the m. tibialis cranialis, the m. flexor digitorum medialis/tibialis and the m. gastrocnemius caput mediale A. The trRemoving the deep fascia and separating the individual muscles reveals some muscles that were previously obscured B. The m.Transecting the m. gastrocnemius and retracting the stumps reveals the m. plantaris on the caudal side of the m. soleus C. The a.When the m. soleus is transected and its stumps are retracted, the more caudal  m. plantaris becomes isolated D. This iWhen the m. plantaris is transected as well, and the m. flexor digitorum medialis/tibialis is retracted cranially, the m. flexor digitorum lateralis/fibularis, which is located lateral (“deep”) to the m. flexor digitorum medialis/tibialis, can be visualized E. In addTransecting the m. flexor digitorum medialis/tibialis and retracting its stumps allows for a better visualization of the m. flexor digitorum lateralis/fibularis F. As alrWhen the foot is approached laterally, the four tendons of the m. extensor digitorum longus, especially the lateralmost one that inserts into the middle and distal phalanges of the fifth digit, are easily recognizable A. The coThe four tendons of the m. extensor digitorum longus can easily be discerned from a dorsal view of the foot B. In addOn the medial view, the paired tendons of the m. tibialis cranialis as well as the tendon of the m. extensor digiti primi  longus can be seen at the level of the proximal retinaculum. The medialmost tendons of the m. extensor digitorum longus to the dorsal sides of the second and third digits can also be discerned. On the plantar side, the m. abductor digiti primi , which starts from the calcaneus and inserts into the plantar side of the proximal phalanx of the hallux, is partly visible. As also seen from the dorsal side, the m. adductor digiti primi  can be seen from the medial view in between the first and second digits. The a. dorsalis pedis superficialis, the a. dorsalis pedis profundus and the v. marginalis medialis pedis can be studied in more detail using the medial approach compared to the dorsal approach.When the plantar side of the foot is studied, the common tendon of the m. flexor digitorum medialis/tibialis can be seen curving around the sustentaculum tali A. This tThe deeper layer clearly shows that the common tendon of the m. flexor digitorum medialis/tibialis splits into four tiny tendons that attach to the plantar sides of the distal phalanges of digits II to V B. The mmDuring this anatomical study, remarkable similarities between the rhesus monkey and a human were observed. Since these similarities stretch beyond anatomy and also apply to physiology, the rhesus monkey plays an important role as a model for humans ,5. HowevWhen the present findings were compared with the literature data on the rhesus monkey anatomy, a few contradictions were noticed. Our dissections revealed that the m. adductor brevis consists of two bellies in the rhesus monkey, whereas the reference works on the anatomy of the rhesus monkey mention the presence of only a single muscle belly ,8. TheseWhen comparing the anatomy of the rhesus monkey with that of common domestic mammals, a number of particularities caught our attention. First, superficial and deep digital flexors, m. flexor digitorum  superficialis and m. flexor digitorum  profundus, respectively, do not exist as such in the rhesus monkey. The m. flexor digitorum (longus) medialis, which has previously been denominated the m. flexor digitorum (longus) tibialis —hence thThe assessment of similarities and differences between the anatomy of the rhesus monkey and that of a human has been completed at the level of the musculature, the circulatory system, including the arteries and veins, and the nervous system. First, some peculiarities regarding the muscles are discussed. In the hip region of the rhesus monkey, the m. gluteus medius is more voluminous compared to the m. gluteus superficialis and the m. gluteus profundus. The former is by far the larger out of the three gluteal muscles in humans. Hence, the term m. gluteus maximus is applied . The deeAt the level of the upper limb, both the m. rectus femoris and the m. biceps femoris consist of two heads in humans and only a single head in the rhesus monkey ,8,16. CoThe m. tibialis cranialis is located on the cranial side of the lower leg. In contrast to humans, in which the m. tibialis anterior is most often single, the m. tibialis cranialis of the rhesus monkey has two muscle bellies ,8. The hAt the level of the foot, the human m. quadratus plantae comprises two heads instead of one in the rhesus monkey ,8,16. ThThe observed differences in the myology of the hindlimb between humans and rhesus monkeys are a reflection of the orthograde posture and associated bipedalism in humans vs. the pronograde posture and associated quadrupedalism in the rhesus monkey . This reBesides differences in musculature, the branching pattern of the arterial system of the rhesus monkey slightly deviates from that in humans. In humans, rhesus monkeys and domestic mammals, the a. profunda femoris branches off the proximal segment of the a. femoralis. In humans, the a. profunda femoris gives off the a. circumflexa femoris medialis. This is a typical trait in domestic mammals . SurprisAt the level of the knee, the human femoral artery continues as the popliteal artery, which gives off the a. genus descendens . A similThe a. saphena is very important in the rhesus monkey, as it plays a pivotal role in the blood supply of the foot, together with the a. tibialis caudalis, which is regarded as the continuation of the popliteal artery in this species . In contThe a. saphena has a venous counterpart called the v. saphena. In fact, the v. saphena is dual. The v. saphena parva in humans drains into the v. poplitea . In mammThe other saphenous vein is the v. saphena magna in humans or v. saphena medialis in mammals, including the rhesus monkey. Irrespective of the species, this vein drains into the femoral vein ,18. The The human terms v. saphena magna and v. saphena parva could be mystifying in the rhesus monkey. Since “magna” means “big, large”, the term v. saphena magna is chosen for that saphenous vein, which is larger in humans. The smaller saphenous vein is termed the v. saphena parva, with “parva” being Latin for “small, tiny”. In the rhesus monkey, however, it is more appropriate to speak of the v. saphena medialis instead of the v. saphena magna and to use the term v. saphena lateralis and not v. saphena parva because, in this species, not the medial but the lateral saphenous vein is larger.The observed differences between the rhesus monkey and humans regarding the conformation of the nerves are rather trivial. In humans, the n. ischiadicus divides into the n. fibularis communis and the n. tibialis just proximal to the knee . In contThis article provides topographical anatomy data on the hindlimb of the rhesus monkey by means of numerous color images taken at consecutive stages of the dissection. This approach allows for gaining insight into the organization of the anatomical structures in several layers, which might be very helpful during, e.g., surgical interventions. In contrast, previous publications on the anatomy of the rhesus monkey present systematic and not topographic anatomy of the hindlimb, thus describing each system, such as the locomotor and vascular systems, separately or fail to present color images.The present work facilitates the translation of experimental data obtained in the rhesus monkey to human medicine and might assist veterinarians during, e.g., wound treatment or surgery. In general, the anatomy of rhesus monkeys shows many similarities to the anatomy of humans and domestic mammals. However, we have described the major anatomical differences between the various species. Since these differences are not numerous and are far from substantial, the rhesus monkey appears to be a valuable model for humans.


A. Snellman, Finland, and TR the Foundation of Märta Donner, Finland. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct Funding statement is: MO Cancer Foundation Finland


In this study, for the first time an analysis of the metal content in extracts obtained from licorice roots grown in the forests of five different regions in Armenia was conducted. Our findings indicated that the concentrations of metals in the extracts did not exceed the permissible limits set by regulatory standards. Furthermore, we investigated the quantitative composition of flavonoids, tannins, and anthocyanins in the licorice roots, which had not been previously studied. Our results revealed that the composition of these substances is significantly influenced by the soil characteristics of the region. To assess the antioxidant properties of the licorice extract, we employed the approach known as the kinetics of competitive reaction method. Our study successfully demonstrated that the extract derived from the roots of the licorice plant, collected from all five regions under study, exhibited notable antioxidant properties.  •Assessment of the concentrations of some heavy metals in licorice roots harvested from five regions of Armenia were done.•Heavy metal concentrations in licorice root were within acceptable limits.•The Kotayk and Tavush regions had the lowest combined heavy metal content.•Investigation of antioxidant properties of licorice roots growing in five regions of Armenia were done.•The Gegharkunik region showed the highest antioxidant activity. The use of such raw materials may lead to a limitation of the use of synthetic materials in these areas. Plants are a natural, valuable source of a certain group of chemical substances that are traditionally used in the production stages of new drugs, cosmetics, food additives, detergents and other products . These sGlycyrrhiza glabra L.) .This work was supported by the Data will be made available on request.No additional information is available for this paper.H.R. Petrosyan: Writing – review & editing, Writing – original draft, Visualization, Supervision, Resources, Project administration, Investigation, Funding acquisition, Data curation, Conceptualization. A.A. Nigaryan: Validation, Formal analysis, Data curation. H.A. Hovhannisyan: Validation, Formal analysis, Data curation. A.M. Soloyan: Validation, Formal analysis, Data curation. V.V. Vardapetyan: Visualization, Project administration, Data curation, Conceptualization. A.I. Martiryan: Supervision, Methodology, Investigation, Conceptualization.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.