Bacteriophages, also known as phages, are viruses that selectively target and infect bacteria. In addition to bacterial dysbiosis, dermatologic conditions such as acne, psoriasis, and atopic dermatitis are characterized by a relative reduction in the abundance of phages and the overgrowth of the corresponding bacteria. Phages often exhibit high specificity for their targeted bacteria, making phage-replacement therapy a promising therapeutic strategy for the control of pathogenic bacteria in dermatologic disease. Novel therapeutic strategies regulating pathogenic bacteria are especially necessary in light of growing antibiotic resistance. In this review, we aimed to review the medical literature assessing phage dysbiosis and therapeutic trials in dermatology. Ultimately, studies have depicted promising results for the treatment of acne, psoriasis, and atopic dermatitis but are limited by low sample sizes and the omission of control groups in some trials. Additional work is necessary to validate the efficacy depicted in proof-of-concept trials and to further determine optimal treatment vehicles, administration mechanisms, and dosing schedules. This review provides the necessary framework for the assessment of phage efficacy in future trials. Corynebacterium diphtheriae have contracted the ability to produce their pathogenic toxin from genetic material acquired by phages [Bacteriophages, also known as phages, are a type of virus that selectively target and infect bacteria . Phages y phages . Additioy phages . Since py phages . It has y phages , whethery phages . This iny phages . Among tTo date, the literature has extensively researched and reviewed the strong association between skin health and a healthy skin microbiome. However, the role of the phageome in regards to skin health has not been as substantially explained. Currently, among the studies that have researched the skin phageome, many have highlighted the lack of a diverse viral database that has been a prominent limiting factor for fully assessing the types of bacteriophages present on the skin ,8. RegarThe unique properties of phages have contributed to the recent re-emergence of interest in phage therapy. Phage therapy involves the administration of phages that selectively target pathogenic bacteria in order to alleviate associated health conditions . AlthougPrevious investigations have assessed microbial and phage compositions among patients with dermatologic disease. Reduced phage concentrations are typically associated with an abundance of the corresponding bacteria, depicting the capacity of phages to suppress their host bacteria . Figure Staphylococcus aureus strains. Furthermore, bacterial alpha diversity has been shown to be reduced in psoriatic lesional skin than in healthy control skin [Affecting almost 3% of the population in the United States, psoriasis is a chronic inflammatory disease characterized by plaques and scaly lesions . Althougrol skin ,15. As prol skin . In a 2020 study, psoriatic lesional skin was obtained from the elbow, forearm, knee, and scalp of sixteen patients with psoriasis . HealthyAcinetobacter phage Presley, Salmonella phage vB_SenS-Ent2, and Bacillus phage SP-10 were the most differentially abundant phages. In addition, the authors observed a significantly greater bacterial alpha diversity in the healthy skin than the lesional skin (p < 0.001) [The authors noted a significantly greater abundance of the top ten most-abundant phage species in the healthy skin compared to lesional skin . Acineto< 0.001) ,18. Acinetobacter phage Presley and Pseudomonas phage O4, and their corresponding bacterial genera. While the family control skin depicted the highest abundance of these two phages, an intermediate and low abundance was demonstrated by the contralateral non-lesional skin and lesional skin, respectively (p = 0.02 for each phage) [p = 0.03 for Acinetobacter; p < 0.001 for Pseudomonas), demonstrating the capacity of phages to suppress the abundance of their host bacteria [The authors further examined two phage species, h phage) . In addibacteria . This fiAcinetobacter baumannii and Pseudomonas aeruginosa, such as wound infections. A 2012 study found that the density of P. aeruginosa decreased in the presence of phages in human skin (ex vivo), suggesting that phage application can effectively alter and correct microbiota compositions [The results of the study conducted by Wang et al.  support ositions . In addiositions , supportInterestingly, the oral consumption of Ganga river water, reported to contain over two-hundred isolates of phages, demonstrated clinical benefits in psoriatic patients who were previously refractory to prescription medication . PatientA Likert Scale from 1–10 was used to categorize the severity of psoriatic lesions. Following the first treatment protocol, the subjects achieved a median improvement of 2.5, although with relapse during the rest period . SimilarIn addition, the phage display of anti-inflammatory peptides has been evaluated for psoriasis-like lesions in mice . Phage dCutibacterium acnes proliferation, further contributing to inflammation and worsening acne severity [C. acnes has long been regarded to contribute to the pathogenesis of acne, recent work has highlighted other bacteria such as Staphylococcus epidermidis [Affecting an estimated 85% of individuals within their lifetime , acne vuseverity . Althougdermidis . NeverthC. acnes and a reduced alpha diversity in comparison to non-inflammatory lesions of acne patients and healthy control subjects [Proteobacteria and Firmicutes overabundance and a reduction in the abundance of Actinobacteria [Staphylococci compared to non-lesional skin, and the proportions directly increased with acne severity. Cutaneous and gastrointestinal microbial dysbioses associated with acne have been previously reported. Acne inflammatory lesions are associated with an overabundance of certain strains of subjects . Similarbacteria . ComedonC. acnes phage abundance have been observed. A 2016 study found a greater relative abundance of C. acnes phages in healthy skin compared to the matched lesional skin of acne patients (p = 0.05) [C. acnes phage abundance and age, suggesting phage abundance may contribute to lower acne vulgaris prevalence among aging individuals. In addition to bacterial dysbiosis between healthy skin and that of acne patients, differences in  = 0.05) , therebyC. acnes bacterial abundance coupled with decreased C. acnes phage abundance in acne patients provides a theoretical framework for the therapeutic replenishment of C. acnes phages. However, it is necessary to note that significant gene conservation has been observed among C. acnes phages, despite the great diversity observed in essentially all other phage populations [C. acnes has evolved strategies to adapt to its cutaneous environment and survive host defenses, such as lipolytic activity, pH regulatory mechanisms, biofilm formation, and the potential activation of dormancy. Such adaptation has resulted in a reduced bacterial diversity, which may have contributed to the observed conservation among C. acnes-infecting phage genomes [C. acnes strains, phage homogeneity poses a therapeutic challenge in that phage host receptor mutations may rapidly confer broad resistance [C. acnes, allows phage-based therapy to be promising for future clinical studies in acne. Increased ulations . C. acne genomes . While bsistance . Still, sistance . FurtherC. acnes-induced inflammation in murine models [C. acnes suspension. While group A received no additional intervention (control), group B received an additional injection with C. acnes at four weeks, and group C received C. acnes and bacteriophages at four weeks. A decreased epidermal thickness and a decreased number and size of microcomedone-like cysts were observed in groups B and C compared to the control group. In addition, the inflammatory nodule size decreased to the greatest extent in group C, depicting the potential efficacy of bacteriophages to reduce C. acnes-induced inflammatory nodules [A 2019 study assessed the effect of dorsal bacteriophage injection on e models . Three g nodules . C. acnes infection in mice, as the antibiotic resistance of C. acnes is growing and poses a challenge for conventional acne treatment [C. acnes followed by phage injection on one side. The C. acnes injection resulted in bilateral inflammatory nodules, although the authors observed a significant reduction in inflammatory lesions on the side injected with the phage. In addition, a significantly decreased expression of the inflammatory marker IL-1β and the apoptotic marker caspase-3 was observed with phage therapy [In addition, a 2021 study assessed the effect of lytic bacteriophage treatment on multi-drug-resistant reatment . Mice we therapy . C. acnes-associated inflammatory lesions, although proper storage is necessary for retaining the lytic activity. In addition, the authors assessed the phage activity of different phage formulations in hydroxyethyl cellulose (HEC) cream . Full lyC. acnes lysis from human skin microflora and formulated them into a cetomacrogol aqueous cream (2.5 × 108 plaque-forming units (PFU) per gram) [C. acnes lysis activity marked a necessary step in creating a topical phage-based acne therapy targeting pathogenic C. acnes. However, the formulation was not directly assessed in human subjects. Similarly, a 2016 study isolated ten phages capable of er gram) . A retaiC. acnes-associated inflammatory nodules in murine models, an additional study evaluated the efficacy and safety of a topical phage formulation among human subjects diagnosed with mild-to-moderate acne [C. acnes with low susceptibility of commensal species. The BX001 cocktail was formulated into an aqueous topical gel. Ex vivo analysis on human skin depicted no irritation effects at any of the studied concentrations, leading the researchers to evaluate the formulations among seventy-five female acne patients stratified into three groups: control, low dose, and high dose. Those receiving the high-dose topical formulation depicted a significantly reduced C. acnes bacterial load compared to vehicle control at day 35 (p = 0.036), although a bacterial reduction was not observed in the low-dose group [C. acnes bacterial load upon the topical application of the phage. However, the authors did not assess the effect of the topical gel on clinical measures of acne, such as inflammatory lesion count or size. Additional research is necessary to assess the clinical efficacy of oral, topical, or intradermal phage formulations among human subjects with acne, although the described studies depict phage replacement to be a promising therapeutic strategy. Whereas the prior studies assessed the effect of phage therapy in the reduction in ate acne . The autse group . While bStaphylococcus aureus [S. aureus growth [Atopic dermatitis (AD) is a chronic inflammatory dermatologic condition commonly present during childhood. Clinically, it presents with patches of intense dryness, scaling, erythema, and pruritus that usually affects the face, neck, scalp, and flexor surfaces of the skin . Typicals aureus . The flas growth . S. aureus growth on the skin [S. aureus bacteria and does not disrupt the beneficial Staphylococcus epidermidis bacteria present on skin. The in vitro experiment involved assessing the effects of S. aureus growth in the presence of SaGU1. During the first 9–13 h, S. aureus levels began to decline in the presence of SaGU1; however, they gradually began to increase after 14 h. This finding suggested that it was probable that S. aureus began to become phage-resistant. A separate experiment involved determining the effects of SaGU1 on S. epidermidis growth, and the results showed that SaGU1 did not affect its growth. Interestingly, the researchers found that when both SaGU1 and S. epidermidis were supplied to S. aureus cultures, the regrowth of S. aureus after 14 h did not occur, and the levels remained suppressed. Therefore, a combination therapy of the phage SaGU1 and S. epidermidis may be beneficial in controlling S. aureus overgrowth. A study conducted in 2020 on atopic mouse models found promising results for the use of a phage in controlling the skin . The stuS. epidermidis or SaGU1 and in combination [S. aureus growth on the skin of atopic mice. However, when the combination therapy was compared to the phage SaGU1 treatment alone, there was no statistically significant difference in terms of effectiveness on S. aureus growth suppression. This suggested that phage therapy alone may be more effective in vivo than in vitro. Overall, these results are extremely promising for the field of phage therapy in treating AD. Although much more intensive research must be performed to determine if phage therapy is clinically useful, this study provided a unique treatment approach that warrants more investigation. During the in vivo phase of the study, atopic mouse skin was treated individually with bination . Each trPhage-based therapeutic strategies appear promising for the treatment of a variety of dermatologic conditions. There are many theoretical attributes of bacteriophages that make them ideal therapeutic candidates for reducing the presence of pathogenic bacteria implicated in disease: phages can be bactericidal, increase in number over time, minimally disturb commensal microflora, depict efficacy against antibiotic-sensitive and antibiotic-resistant bacteria, can potentially disrupt bacterial biofilms, and exhibit low toxicity . HoweverCertain phage characteristics predict their therapeutic success. First, in order to allow optimal bacterial lysis, virulent phages must be selected in contrast to temperate phages. Similarly, phages characterized by a poor killing potential against desired bacteria are sub-optimal for therapy. Toxin-carrying phages should be avoided in addition to those depicting a high transduction potential . Lastly,Additional factors mediate phage-based treatment efficacy, including the phage-to-target bacteria ratio, environmental conditions, accessibility to target bacteria, administration mechanisms, and bacterial resistance . First, Secondly, pH has been shown to affect phage titers, with decreasing pH associated with reduced titers. In addition, phages cannot freely diffuse across membranes and thus require a delivery method to reach target cells and bacteria. Administration techniques include oral, topical, intraperitoneal, intravenous, and intranasal administration . HoweverPseudomonas aeruginosa with resistant O-antigen deletion mutants [Lastly, experimental data has demonstrated the presence of phage resistance in up to 80% of studies targeting the intestinal microbiome . Bacteri mutants . The phaUltimately, phage therapy is not without limitations, including the development of resistance, environmental factors, and the necessary consideration of the optimal administration mechanism. Perhaps such limitations have contributed to the fact that only a few countries have accepted the use of therapeutic phages in humans thus far . HoweverBacteriophage dysbiosis has been observed on the lesional skin of psoriasis and acne patients. Experimental confirmation of the suppressing capacity of phages on their corresponding bacteria suggests that phage dysbiosis may contribute to the microbial dysbiosis characteristics of each dermatologic condition. In addition, phage deficiency with associated bacterial abundance suggests that phage replacement, either topically or orally via phage “cocktails,” may be an effective therapeutic strategy to mitigate the phenotypic signs and symptoms of disease. Few studies assessing phage-based therapies have been promising for the treatment of psoriasis, acne, and atopic dermatitis, both in murine models and human subjects. However, the current evidence is limited by low sample sizes and the omission of controls in some cases. Future research is necessary to assess the efficacy of phage replacement in large scale studies in addition to determining the optimal treatment vehicles, administration mechanisms, and dosing for particular purposes.


Xanthomonas euvesicatoria. Phages were isolated from the rhizosphere of tomato plants with symptoms of bacterial spot. The plaque morphology of all isolates was determined on a X. euvesicatoria lawn via a plaque assay. Three of the isolates were attributed to the family Myoviridae based on TEM micrographs. All phages showed good long-term viability when stored at 4 °C and −20 °C. Three of the phage isolates possessed high stability at very low pH values. Fifty-five-day persistence in a soil sample without the presence of the specific host and a lack of lytic activity on beneficial rhizosphere bacteria were found for the phage isolate BsXeu269p/3. The complete genome of the same isolate was sequenced and analyzed, and, for the first time in this paper, we report a circular representation of a linear but circularly permuted phage genome among known X. euvesicatoria phage genomes.Bacteriophages have greatly engaged the attention of scientists worldwide due to the continuously increasing resistance of phytopathogenic bacteria to commercially used chemical pesticides. However, the knowledge regarding phages is still very insufficient and must be continuously expanded. This paper presents the results of the isolation, characterization, and evaluation of the potential of 11 phage isolates as natural predators of a severe phytopathogenic bacterium— Xanthomonas euvesicatoria pv. euvesicatoria (formerly known as Xanthomonas euvesicatoria), X. euvesicatoria pv. perforans (formerly known as Xanthomonas perforans), Xanthomonas hortorum pv. gardneri (formerly known as Xanthomonas gardneri), and Xanthomonas vesicatoria) has cyclic peaks. It is an economically important disease and a severely limiting factor for fruit yield in these crops worldwide [Bacterial spot in tomato and pepper  that infect tomato and two species (X. vesicatoria and X. euvesicatoria) that infect pepper have been described in Bulgaria. The natural population of pathogens is heterogeneous by symptoms, species, phenotypic and genotypic characteristics, pathotype, and races [To date, three species  [X. vesicatoria (an Inoviridae phage [Myoviridae phage [Podoviridae phage) [X. perforans (unidentified phages) [X. gardneri  phage) [Xanthomonas phages, with only 134 fully sequenced genomes compared to 133 for Escherichia coli phages, 543 for Acinetobacter phages, 231 for Erwinia phages, etc. (NCBI). Out of these 134 fully sequenced genomes, only five are from Xanthomonas phages associated with bacterial spot xanthomonads: one X. euvesicatoria phage (access. number NC_054460); one X. campestris pv. vesicatoria phage (access. number MH206183); and three X. vesicatoria phages .Deep and extensive studies on the nature of bacteriophages and their role in combating severe bacterial plant diseases have been carried out in the last 20 years or so. Hopeful results regarding phage biocontrol have been reported for several devastating bacterial plant diseases, such as: potato soft rot , bacteri species . There hicatoria ,23,24; p Serbia) ,26; phagae phage , a Myoviae phage , and onee phage) ; phages  phages) ; and pha) phage) . MoreoveRalstonia solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. vesicatoria, X. euvesicatoria, and X. perforans [X. euvesicatoria phage KΦ1 to different pH values, storage temperatures, and UV radiation in vitro has been reported [X. euvesicatoria, X. vesicatoria, and X. gardneri [It is known that the rhizosphere and leaf surfaces are the main areas for the application of phage-based biopesticides. Studies reporting phage survival after application in soil and the phyllosphere have been reported for phages infecting erforans ,32,33,34erforans ,23,35. Treported . The samgardneri . PerhapsX. euvesicatoria-specific bacteriophages and the determination of several main and specific characteristics concerning their biology and biocontrol potential. The aim of this paper was the isolation of X. euvesicatoria strains, including the type strain, regardless of their pathotype, race, and source of isolation (host and region). The strains of the remaining four species of phytopathogenic bacteria  were resistant to the studied phages  around the center. Slight differences were observed only in plaque dimensions. The diameter of the clear center ranged from 1 to 3.5 mm, with an average value of 2.3 mm, and the diameter of the whole plaque  ranged from 3 to 5 mm, with an average value of 4 mm . Myoviridae family (phages with long contractile tails), based on the classification of Ackermann [The transmission electron microscopy of the three selected phages showed that the virions had icosahedral capsids (approx. 50 nm) and long contractile tails (approx. 100 nm) . These rckermann  and Bradckermann .Xanthomonas phages such as KΦ1 (KPhi1) . The fiPure phage lysates were stored under the chosen temperature conditions for a period of 11 months. The phage titers at 0h were determined and used for subsequent comparison with the titers obtained during the storage period. The concentration of viable phage particles was determined by point testing  . The obtX. euvesicatoria strains 105t and 269p) was investigated. We found that an inhibitory effect was only observed when applying undiluted buffers with pH 13.0 for both strains, as well as with pH 2.0 for strain 105t , and 7.5, respectively. Incubation at pH 9.0 and 11.0 caused a slight titer decrease of about 1 lg units only for phage isolate BsXeu269p/3 . A stron10 pfu/mL). No viable phages were detected on the 62nd day.The ability of phage isolate BsXeu269p/3 to retain its lytic activity in soil was studied  mainly related to the nitrogen cycle in the soil. For this purpose, the total number of these groups was previously determined in the soil sample used and is presented in ectively .X. euvesicatoria. This study was a continuation of our previous work [Bacterial spot disease in tomato and pepper plants is spread worldwide especially in countries with warm and humid climates. Disease management is difficult due to increasing bacterial resistance to conventionally used copper-based pesticides ,11. Therous work .X. euvesicatoria  and then characterize them according to parameters that are relevant for their further study, with a view to future application as biocontrol agents. In this paper, 11 bacteriophages isolated with the bacterial host X. euvescatoria are reported. The host range of four of them was determined, and the results showed that they could infect all strains of the species, regardless of the host and the region from which they were isolated. According to the widely accepted concept of host range determination, these phage isolates could be defined as broad-host-range phages [X. euvesicatoria in Serbia [X. euvesicatoria has been mainly reported as a bacterial spot pathogen on pepper [X. euvesicatoria-specific phages (XESP). This difference in the source of XESP isolation could be explained by the fact that in Bulgaria this pathogen infects tomatoes too [X. vesicatoria and X. gardneri [In this study, the main objective was to isolate and propagate phages specific to e phages . To our n Serbia ,26. The n Serbia . So far,toes too . In our gardneri .X. euvesicatoria phages by Gasic et al. [X. euvesicatoria, as member of the Xanthomonas genus, formed extracellular polysaccharide xanthan [Erwinia amylovora phage f-Ea1h [Pseudomonas aeruginosa [Escherichia coli K12 [Rhizobium trifolii [Xanthomonas phages forming clear plaques without a halo have also been reported [The morphology of plaques is an essential characteristic. All our phage isolates formed approximately identical plaques, as the differences were observed only in plaque size. The formation of clear plaques is typical for lytic phages . The prec et al. . X. euve xanthan , and thee f-Ea1h , Pseudomruginosa , Eschericoli K12 , and Rhitrifolii . Howeverreported .Myoviridae family had relatively small capsids (approx. 50 nm), but the plaques they formed were relatively big (4 mm). Some publications have claimed that the plaque size and capsid size are inversely related. Phages with a smaller capsid formed a large plaque in . The autMyoviridae family, such as T4-like and P1-like viruses [X. vesicatoria phage phiXaf18 [Although the genome of BsXeu269p/3 was best represented as a circular molecule, this phage has a physically rather linear, although circularly permuted genome, as in other representatives of the  viruses . CirculaphiXaf18 . The iniphiXaf18  were invXanthomonas phages [Xanthomonas arboricola phages [Different strategies have been studied for the long-term storage of pure or purified phage lysates , but the efficacy of each method varies and depends on the phages themselves . Generals phages ,29,53. Ha phages . Deep fra phages , but we Xanthomonas phages is between 5.0 and 11.0, with a few exceptions whereby phages have shown resistance to lower pH values (pH 4.0). To a large extent, our results were consistent with those reported by the latter authors. However, data demonstrating phage tolerance to pH 2.0 could also be found for other Myoviridae phages specific to P. aeruginosa и E. coli [Among the main strategies for the application of phage-based biopreparation in agriculture is administration via the rhizosphere or phyllosphere of plants. The phyllosphere is the part of the plant that is exposed to the action of a number of environmental factors such as UV radiation, wind, rain, and drought. These factors have been reported to affect the survival of phages on the leaf surface, thereby reducing their efficiency ,35. In t E. coli .X. euvesicatoria. The experiment was thus designed to determine the ability of the phage to remain viable under the conditions described. Furthermore, X. euvesicatoria is not a soil-borne phytopathogenic bacterium, so it can only persist in the soil for a limited period of time. Thus, we conducted an experiment aiming to more closely simulate natural soil conditions. The results obtained gave us reason to conclude that this phage would be suitable for soil application, since under these conditions it remained viable for 55 days. Similar results have been previously reported for several phages specific to E. amylovora, R. solanacearum, X. perforans, X. euvesicatoria, and X. oryzae, which can migrate from roots to the aerial parts of plants when applied to the rhizosphere [Another important feature of phages intended for soil application is their long-term persistence. In this study, we selected the phage isolate BsXeu269p/3, which was inoculated into a soil sample under in vitro conditions in the absence of its host, zosphere ,32,34,55Rhizosphere microorganisms play a key role in plant growth. Therefore, it is important that their structure and function are not negatively affected during the application of plant disease control practices in agriculture. Copper-based preparations kill not only the target phytopathogenic bacteria, but also many microorganisms that are beneficial to the plant. Bacteriophages are considered safe in this regard due to their high specificity towards the target microorganism. However, before a bacteriophage is proposed as a biocontrol agent, its impact on non-target microbiota should be investigated to demonstrate its safety. In our study, we demonstrated the safety of one phage isolate (BsXeu269p/3) against four groups of rhizosphere bacteria. X. vesicatoria, X. euvesicatoria, X. gardneri, and P. syringae pv. tomato) were used in this study  were used as host microorganisms for bacteriophage isolation , X. vesicatoria (14 strains), X. gardneri (3 strains), and P. syringae pv. tomato (1 strain)) and five types of cultures, shown in Pure bacteriophage cultures were obtained in LB broth with their target bacterium, according to the procedure described previously by Kizheva et al. , and stoX. euvesicatoria strains 105t and 269p were used as host bacteria for 11 phage isolates, 5 of which were designated as BsXeu105t and 6 as BsXeu269p. The analysis was performed by measuring the diameter of the plaques and the determination of the type of plaque center and edges.The morphology of the plaques produced by the phages on the surface of the semi-solid agar medium inoculated with the target bacterium was determined by DAOPA. 9 pfu/mL) was used for preparing the negatively stained formvar-coated grids for observation by TEM [The virion morphology of three phage isolates  was investigated by transmission electron microscopy (TEM). The bacteriophages were propagated on a semi-solid agar medium (Nutrient Agar (NA), Merck KGaA, Darmstadt, Germany) by DAOPA along with their target bacterium. Fresh phage suspensions were prepared in phage buffer , accordin by TEM . The obs® Syringe Filter 28 mm, pore size 0.22 μm, Sartorius, 37079 Goettingen, Germany) to completely remove residual bacterial cells; then, it was used for the extraction of phage genetic material by a NORGEN Phage DNA isolation Kit . The sequences were obtained after whole-genome sequencing by Illumina NovaSeq PE150 in Novogene (Novogene (UK) Company Limited, Cambridge, CB4 0FW, United Kingdom).Fresh phage suspension was obtained after the propagation of the phage isolate BsXeu269p/3 with its specific host on NA medium by DAOPA. Five milliliters of phage buffer  was pourwww.clcbio.com, accessed on 12 May 2022). The genome annotation was performed manually and using the online platform RAST [Raw sequences were assembled into contigs (contig 1 and 2) using Unicycler  . Two priorm RAST .The determination of phage survival at different storage temperatures (4 °C and −20 °C) was carried out using pure phage lysates, obtained after propagation in LB medium. Glycerol solution  was used as a cryoprotectant for phage lysates stored at −20 °C. Phage titers were determined by a spot-testing assay, and the determination was performed at various time intervals for the different phage isolates. The experiments were carried out in triplicate.X. euvesicatoria strains 105t and 269p were used as host strains. In order to obtain a pure phage suspension, 10 mL of each buffer solution was poured on the surface of the obtained phage cultures in petri dishes and incubated on a rotary shaker at room temperature for 1.5 h. At this step, only viable phage particles passed from the medium surface into the buffer solutions. The resulting phage suspensions were filtered through a membrane filter (pore size 0.22 μm) to completely remove the residual bacterial cells. The effect of the pH on phage viability was evaluated based on the difference in phage titer at 0 h, and that at 48 h was determined by a spot-testing assay, as follows: a series of tenfold dilutions of the phage suspensions were prepared, and then 10 µL of each dilution was spotted on the surface of the semi-solid agar (NA) medium along with bacterial hosts, followed by cultivation at 28 °C for 24 h. The sensitivity of the host bacteria to different pH values (controls) was determined using the same buffer solutions but in the absence of phage particles. A series of tenfold dilutions of the buffer solutions were prepared, and 10 µL of each dilution was spotted on the solid agar medium surface inoculated with the host bacterium (100 µL–108 cfu/mL). All experiments were performed in duplicate. The design of the analysis is illustrated in Basic buffer solution with pH 5.92 was prepared as described by Msimbira et al. . It was The soil sample was collected from the rhizosphere of a healthy tomato plant in 2018 from a private vegetable garden in the village of Novo Panicharevo, Primorsko municipality, Burgas region, southeastern Bulgaria. The sample was stored in a sterile plastic jar at 4 °C and immediately transported to the laboratory, where it was analyzed. X. euvesicatoria-specific phages. Pure phage lysate was obtained for the selected phage isolate (BsXeu269p/3) in LB medium. The initial titer (1010 pfu/mL) of the resulting phage culture was determined by a spot-test assay. Ten milliliters of the phage lysate was added to a 30 g soil sample (in triplicate) in a sterile plastic jar and incubated at room temperature for a period of 62 days. The persistence of the viable phage particles was studied by DAOPA (host—X. euvesicatoria strain 269p), and the phage titers on the 55th and 62nd day of incubation were compared to the initial phage titer.To determine the time limit of phage survival in soil, we used a soil sample free of naturally occurring −7) of the soil suspension in saline were prepared, and an aliquot of 100 µL of each dilution (in triplicate) was plated on the surface of the appropriate culture media  of beneficial rhizosphere bacteria  was calculated in a native soil suspension prepared in saline . The media, methods, and cultivation conditions applied in the analyses are listed in re media . The num10 pfu/mL was used. The analysis was carried out by two methods—DAOPA and MPN. We used DAOPA to determine the ability of the phage isolate to form clear plaques on a lawn of previously enriched soil heterotrophs (in NAII medium) and nitrogen-fixing bacteria (in nitrogen-free broth medium)  or NFM agar (0.45% agar) for heterotrophs and nitrogen-fixing bacteria, respectively, was poured onto the surface of the respective agar media in previously prepared petri dishes. The phage lysate was diluted up to 10−7, and an aliquot of 10 µL of each dilution was spotted on the surface of the prepared double agar plates, which were cultivated at 28 °C for 24 h. The formation of clear plaques after cultivation indicated the presence of heterotrophic and nitrogen-fixing bacteria susceptible to the phage isolate. The MPN method was applied to study the ability of the phage isolate to reduce (by lysis) the amount of soil microorganisms in liquid media (−7), 100 µL pure phage lysate (1010 pfu/mL), and 10 mM CaCl2 was added to each test tube containing liquid media. To determine the influence of the phage isolate BsXeu269p/3 on the beneficial rhizosphere microorganisms, a pure lysate with a titer of 10 medium) . A mixtuid media . A mixtuX. euvesicatoria phages. This, along with the other characteristics possessed by this phage , makes it a potential candidate as a biocontrol agent against bacterial spot disease in tomato and pepper.Our study emphasized the necessity of studying bacteriophages as promising candidates for combating bacterial infections. We believe that our results for the genome organization of phage BsXeu269p/3 and the virion morphology of two more phages will contribute to the study of phage biology in general. Moreover, for the first time in this paper, we provided evidence of the circular permutation of a linear genome among the known


The endocannabinoid system is involved in physiological and pathological processes, including pain generation, modulation, and sensation. Its role in certain types of chronic orofacial pain (OFP) has not been thoroughly examined. By exploring the profiles of specific salivary endocannabinoids (eCBs) in individuals with different types of OFP, we evaluated their use as biomarkers and the influence of clinical parameters and pain characteristics on eCB levels. The salivary levels of anandamide (AEA), 2-arachidonoyl glycerol (2-AG), and their endogenous breakdown product arachidonic acid (AA), as well as the eCB-like molecules N-palmitoylethanolamide (PEA) and N-oleoylethanolamide (OEA), were assessed in 83 OFP patients and 43 pain-free controls using liquid chromatography/tandem mass spectrometry. Patients were grouped by diagnosis: post-traumatic neuropathy (PTN), trigeminal neuralgia (TN), temporomandibular disorder (TMD), migraine, tension-type headache (TTH), and burning mouth syndrome (BMS). Correlation analyses between a specific diagnosis, pain characteristics, and eCB levels were conducted. Significantly lower levels of 2-AG were found in the TN and TTH groups, while significantly lower PEA levels were found in the migraine group. BMS was the only group with elevated eCBs (AEA) versus the control. Significant correlations were found between levels of specific eCBs and gender, health-related quality of life (HRQoL), BMI, pain duration, and sleep awakenings. In conclusion, salivary samples exhibited signature eCBs profiles for major OFP disorders, especially migraine, TTH, TN, and BMS. This finding may pave the way for using salivary eCBs biomarkers for more accurate diagnoses and management of chronic OFP patients. Chronic orofacial pain (OFP) is a debilitating condition that is associated with the structures innervated by the trigeminal nerve . It is one of the most common pain conditions, with a reported prevalence of 7–11% in the general population . The difDue to the nature of pain as a sensory experience that cannot be directly quantified or measured , subjectSensation to the orofacial tissues, such as teeth, facial skin, TMJ, and adjacent musculature, is mainly supplied by branches of the trigeminal (V) nerve . Pain paN-arachidonoylethanolamide (anandamide) (AEA) and 2-arachidonoyl glycerol (2-AG)); and (3) enzymes, which are responsible for the biosynthesis/inactivation of the ligands. Furthermore, AEA is a member of the N-acylethanolamines (NAEs) family, which includes the eCB-like compounds N-palmitoylethanolamide (PEA) and N-oleoylethanolamide (OEA) [In this context, recent research found that the endogenous cannabinoid (endocannabinoid (eCB)) system is essential in pain pathophysiology , and thede (OEA) . N-acylphosphatidylethanolamine-phospholipase D (NAPE-PLD) catalyzes AEA biosynthesis, whereas 2-AG is catalyzed by sn-1-specific diacyl-glycerol lipase (DAGL). The released eCBs are retrieved by a membrane transporter. AEA is degraded by fatty acid amide hydrolase (FAAH), whereas monoglyceride lipase (MAGL) is the main 2-AG hydrolase [eCBs are synthesized and released locally by enzymatic cleavage of membrane phospholipids in response to physiological and pathological stimuli . N-acylpydrolase . 2-AG isydrolase . AEA alsydrolase . ECS comydrolase , which bydrolase . Based oydrolase , it is rWhereas blood serum/plasma is the most frequent source of measurable biomarkers, saliva has many advantages over blood as a collectible bio-fluid, such as being easy to collect in a non-invasive manner and safer to handle. Many substances enter saliva from the blood via intercellular spaces due to transcellular or paracellular diffusion, and most substances found in the blood are also present in saliva, reflecting physiological and pathological states, making it a useful source of biomarkers for pain research .A recent study conducted by our group demonstrated significantly reduced levels of salivary eCB in chronic OFP disorders, which were categorized by etiology  . This avA total of 126 individuals participated in the current study, 83 of which had OFP, including TMD (25.3%), migraine (28.9%), TTH (6%), PTN (13.4%), TN (13.4%), and BMS (8.5%). A total of 4.8% were defined as “others”. Of the 126 participants forming the cohort, 83 (66%) had OFP, while 43 (34%) were pain-free and formed the control group. Of the OFP group 21 (25.3%) had TMD, 24 (28.9%) had migraine, 5 (6%) had TTH, 11 (13.4%) had PTN, 11 (13.4%) had TN, 7 (8.5%) had BMS, and the remaining 4 (4.8%) were defined as “others”.The primary headaches group represents the sum of migraine  and TTH, while the neuropathic group included PTN, TN, and BMS. Considering the vast amount of data collected and the many comparisons performed between the clinical factors and patient pain characteristics for each of the five measured eCBs, we decided to only present the statistically significant results. It is important to note that for some of the diagnoses, e.g., BMS and TTH, there were not enough patients for within-group comparisons.The limited understanding of the etiology and pathogenesis of OFP disorders, the subjective nature of current pain assessments, and the limited efficacy of existing treatment options highlight the urgent need for objective data to assist with chronic pain evaluation and management. The identification of mechanistic biomarkers is crucial since it would not only improve our understanding and ability to diagnose pain disorders accurately but also facilitate the development of disease-modifying drugs . Over thOur data demonstrated significantly lower salivary PEA levels in the migraine group compared with the controls. No other significant differences between these groups were found. PEA, an endogenous fatty acid amide signaling molecule, is synthesized on demand as a protective response to tissue injury or stress as part of homeostatic mechanisms. It has anti-inflammatory, pain relieving, and neuroprotective actions . PEA’s dOne of the most important anti-inflammatory effects of PEA is the inhibition of mast cell activation , which iN-acylethanolamine acid amidase (NAAA), which catabolize PEA and AEA at significantly different rates [Interestingly, in contrast to our findings, Sarchielli et al.  found thnt rates , it is pnt rates .Our data also demonstrated low salivary 2-AG levels in the TTH group compared with the controls. Although TTH is the most prevalent type of headache, its pathophysiology remains unclear . InterreWe also found significantly lower levels of salivary 2-AG in the TN group. TN is a chronic neuropathic condition that affects one or more divisions of the trigeminal nerve . Devor eN-arachidonoyl-dopamine. Additionally, the intrinsic efficacy of AEA on TRPV1 is lower than at the CB1 receptor, implying that in the presence of CB1, AEA action on TRPV1 may be attenuated [Surprisingly, we found an elevation in salivary AEA levels for BMS compared with the controls. No other significant differences between these groups were found. BMS is a chronic neuropathic pain disorder characterized by an oral mucosal burning sensation and is frequently associated with xerostomia and dysgeusia . The etitenuated . AEA is tenuated . Indeed,tenuated . Therefore, despite AEA being a known anti-nociceptive eCB and its stimulation of cannabinoid receptors producing analgesic effects, it may act as an antagonist in the presence of full cannabinoid agonists . Taken together, we hypothesize that the significant elevation of salivary AEA in BMS with no concomitant elevations of full eCB agonists, such as 2-AG, reflects a faulty mechanism of the ECS in these patients. The elevated AEA accompanied by altered epithelial tongue cell eCB receptor expression  implies In addition to the correlations between specific pain disorder subgroups and salivary eCB levels, we also found a correlation between ECS activity and patient characteristics in three pain disorder groups: TN, PTN, and migraine. There were significantly lower salivary AA levels in women than men in the PTN group and this trend was also noted for OEA and PEA levels in the migraine group. These findings are consistent with Cupini et al. , who detInterestingly, our data indicated a significant negative correlation between the VPS and salivary OEA and PEA levels in the migraine group. This is congruent with previous studies, where chronic administration of PEA reduced pain behaviors and counteracted spinal neuronal hyper-excitability in murine models of persistent pain ,41.HRQoL reflects the perceptions and reactions of an individual to their health status and the non-medical aspects of their lives and assesses their psychological functioning and, to a smaller extent, physical functioning . We founWe found that a BMI above 30 was significantly related to higher salivary 2-AG levels in the migraine group. This is consistent with previous studies, where levels of circulating 2-AG positively correlated with BMI, total body fat, and intra-abdominal adipose tissue ,43. ThisIn the TN group, a positive correlation was found between sleep awakenings and salivary AEA, OEA, and PEA levels. Significant OEA elevation was shown in human CSF after 24 h of sleep deprivation . It is pFinally, our data demonstrated a significant positive correlation between pain duration and salivary AA levels in the TN group. This may have been due to pain-management-related symptom relief over time. This assumption is strengthened by the inverse correlation between pain intensity and salivary eCB levels .There were some important limitations to this carefully designed study. First, we only measured salivary eCB levels from samples taken at a single point in time, gathering samples over more time points may reveal other important information, such as correlations between treatment outcomes and eCB levels or their correlation with long-term clinical symptoms. Second, there were several therapeutic treatments with different efficacies, which could have affected the salivary eCBs concentrations. Third, serum eCB levels were not measured in our study. Thus, a correlation between salivary and circulating eCBs could not be evaluated. Fourth, we had relatively small sample sizes for each subgroup, which limited the comparative statistical analysis.The study was approved by the Ethical Committee of Hadassah Medical Center, request no. 0662-17-HMO. All data were fully anonymized; informed consent was waived according to the ethical committee’s instructions. The medical records of 83  OFP patients meeting our inclusion criteria attending the Orofacial Pain Clinic at the Hebrew University-Hadassah School of Dental Medicine between 2017 and 2018 were reviewed. Forty-three  pain-free participants formed the control group.Inclusion criteria: over 18 years of age, definite diagnosis of chronic OFP for at least 3 months according to the IHS or ICOP ,48, and Exclusion criteria: background illnesses, including cancer or diseases affecting the salivary glands, such as Sjögren syndrome; alcohol use; patients who did not sign the consent form; and patients whose saliva sample was unusable, e.g., too foamy or bloody.The pain patients were divided based on their diagnoses:Temporomandibular disorders (TMD) according to the diagnostic criteria for temporomandibular disorders (DC/TMD) , which aPrimary headaches , including migraine  and tension-type headache (TTH), according to the ICHD-3 . Trigeminal neuralgia (TN) , which iPost-traumatic trigeminal neuropathy (PTN) , which iBurning mouth syndrome (BMS), which is a chronic pain condition characterized by a moderate-to-severe sensation of burning from the oral mucosa, especially from the dorsum of the tongue with no clinical signs . TN, PTN, and BMS formed the “neuropathic” group. Other diagnoses that were rare and non-specific (8 patients) formed the “others” group.Primary and resultant data were recorded on the standard intake form used in our clinic ,53. DemoUnstimulated saliva was collected for 10 min as described previously ,59 into Saliva samples were centrifuged at 3500× g for 10 min at 2 °C to remove insoluble materials, cell debris, and food remnants. The supernatant fraction was aliquoted into polypropylene tubes and immediately stored at −80 °C. The running order of the samples was cycles of 2 samples from pain patients with the same diagnosis and then a control sample with the same gender and general BMI. 3:MeOH  and then washed with ice-cold chloroform three times. The samples were then dried under a thin stream of nitrogen and reconstituted in MeOH. Analysis using LC-MS was performed on an AB Sciex  Triple Quad 5500 Mass Spectrometer and a Shimadzu  UHPLC System, while the liquid chromatographic separation was acquired via a Kinetex (Phenomenex) column . Sample levels of AEA, 2-AG, AA, PEA, and OEA were measured against standard curves and then expressed as fmol/mg protein.The extraction, purification, and quantification of saliva eCBs were performed using stable isotope dilution liquid chromatography/tandem mass spectrometry (LC-MS/MS) as previously described . BrieflySPSS version 25 software was used for all calculations. To examine the differences in eCB levels for nominal and categorical background variables, t-tests and one-way analysis of variance were performed, and when significant differences were found, additional post hoc Scheffe tests were performed. A Spearman coordinator was used to examine the specific categories that made up the differences. The differences between the eCB types and specific background variables were examined for each diagnosis using the Kruskal–Wallis test.Our findings suggested that the ECS played a significant role in the pathogenesis of OFP disorders. In addition, the non-invasively gathered salivary samples exhibited signature eCB profiles for prominent OFP disorders. Therefore, the profile of salivary eCBs and eCB-like molecules may be used as biomarkers to aid in the diagnosis and management of these patients. Future research is needed to evaluate whether salivary eCB levels correlate with serum levels in chronic OFP disorders, to understand the underlying mechanisms of altered ECS found in these patients, and to evaluate the therapeutic use of cannabinoids possibly as part of a personalized medicine approach for the management of chronic OFP disorders.


Strike action in healthcare has been common over the last several decades. The overarching aim of this systematic review was to synthesise and analyse the empirical literature that examines the impact of strike action on patient morbidity, that is, all patient outcomes except mortality. After conducting a search and apply eligibility criteria, 15 studies were included in this review. These articles included a variety of outcomes from hypertension control to rates of chlamydia. Strikes ranged from 13 to 118 days, with a mean strike length of 56 days. A textual narrative synthesis was employed to arrange studies by whether they had a positive, mixed or neutral or negative impact on patient morbidity. Results suggest that strike action has little impact on patient morbidity. The majority of studies reported that strike action had a neutral or mixed impact of strike action on patient morbidity. One study reported positive outcomes and three studies reported negative outcomes, however in both cases, the impact that the strike had was marginal. Strike action is widely debated with a major concern related to patient outcomes, including patient morbidityStrike action has little impact on patient morbidity, almost all studies reported no adverse outcomesStrike action by healthcare staff can be carried out safely in relation to patient morbidity Strikes have occurred on almost every continent, for a range of reasons. They have been carried out over a matter of hours to hundreds of days. While healthcare strikes raise a range of issues, one of the most pressing that is almost always raised relates to the impact that the action could have on patient wellbeing. That is, most debates centre on the impact that strike action could have on patients, with those arguing both for and against such action citing patient safety as a major concern.Over the years there has been a growing body of evidence that has examined the impact of strike action on the health and wellbeing of patients. The majority of this evidence has examined patient mortality, with evidence suggesting that generally, strikes do not significantly change patient mortality in‐hospitalThe overarching aim of this review is to synthesise and analyse the empirical literature that examines the impact of strike action on patient morbidity, that is, all patient outcomes except mortality. This reviews seeks to (1) understand if strike action has an impact on morbidity and if so (2) what factors related to the strike, or the health of patients in particular impact these outcomes.22.1A systematic review was employed to identify and synthesise all relevant literature in relation to the above research questions. PRISMA and ENTREQ reporting guidelines were followed.2.2The following electronic databases and time periods were searched: EMBASE (1980–2021), MEDLINE (1946–2021), CINAHL (1982–2001), BIOETHICSLINE (1972–1999), EconLit (1969–2021), WEB OF SCIENCE (1960–2021). In addition, grey literature was searched through OPEN GREY, and SIGMA REPOSITORY. Search terms were developed to capture the core concepts related to the form of intervention we were interested in  and the populations in question . The final search terms were: strike OR ‘industrial action’ OR ‘industrial dispute’ OR ‘collective action’ AND doctor OR physician OR clinician OR ‘medical practitioner’ OR nurs* OR ‘health profession*’ OR healthcare OR ‘health care’ OR ‘pharmac*’ OR ‘dentist’ OR ‘midwi*’ OR dieti* OR ‘occupational therap*’ OR ‘paramed*’ OR ‘physiotherap*’ OR ‘radiograph*’ OR ‘psycholog*’ OR ‘health worker’ OR ‘hospital’. There were no publication dates or language restrictions. Where complete data for a relevant outcomes was not available we contacted authors to request data. In addition, we conducted a manual search of the reference lists of eligible studies.2.3The initial search returned 5728 results, which were imported into Endnote where duplicates were removed. This left 4415 articles. The title and abstract of these articles was scanned and articles not meeting the inclusion criteria were removed. After the initial screen, 392 articles remained and a second full‐text screen was undertaken and reference lists were searched. A further four papers were found and all 396 articles were assessed against the below eligibility criteria, leaving 15 articles  or patient mortality.They examined patient mortality during strike action.Papers were excluded if:2.4Data from the included studies was extracted by RE, checked by SMW and categorised according to the source, country of where the research took place, study aims and objectives, research methods/design, the context of the study, nature of the strike, main outcomes, and quality appraisal scores and issues .2.6Studies were combined to summarise descriptive statistics of the study characteristics, followed by a textual narrative synthesis. This approach arranges disparate study types into more homogenous sub‐groups which aids in the synthesising of different types of evidence and in this case, answering research questions which can be informed by multiple methodological approaches.3The 15 articles included measured a variety of outcomes from hypertension control to rates of chlamydia. The articles also included substantial geographic diversity with studies from Europe, North America, Africa and Asia. Six studies examined strikes by doctors, four examined strikes by nurses, with the remainder of studies examining strikes by ambulance staff, ‘non‐professional employees’, government employees and multiple healthcare staff from a mental health centre. Strikes ranged from 13 to 118 days, with a mean strike length of 56 days. These results are summarised in Table 3.1Thirteen of the papers included in this review were reviewed against the criteria set out in the NOS. Two papers were excluded as they did not utilise observational designs.3.2Papers were categorised into whether they had a negative, mixed or neutral or positive impact on patient morbidity. Studies were categorised as having a negative impact when they reported worse patient outcomes during a strike. Studies were categorised as neutral when they reported no substantial impact on patient outcomes during a strike. And studies were categorised as positive when they reported an improvement in patient outcomes during a strike. Studies were categorised as mixed when they reported a mixture of positive, negative or neutral results. There were three studies that reported a clear negative impact. These studies varied substantially. There were four studies that reported a mixed impact of strike action. Amongst these studies, three were conducted in Canada and one in Israel. One Israeli study reported a positive impact of strike action. There were no obvious patterns that linked these outcomes to the nature of the strike, for example, the length of strike, when or where the strike occurred.3.3There were three studies that reported a clear negative impact. These studies varied substantially. These were carried out in Kenya, Denmark and the UK. One involved ambulance staff, the two involved nursing staff. The strike in Kenya lasted 100 days, the strike in Denmark lasted 60 days, while the strike in the UK last 35 days. The studies examined the impact on immunisation services, diabetic control and more general outcomes from a day hospital.Taking a closer look at the first of these studies, Njuguna3.4There were four studies that reported a mixed impact of strike action. Amongst these studies, three were conducted in Canada and one in Israel. Strikes ranged in length from 31 to 118 days. Each study examined a different group of workers; doctors, nurses, ‘non‐professional’ health workers and sexual health programme workers. The outcomes examined include hypertension control, mental health, caesarean birth rates and prevalence of Chlamydia.In one of the earliest studies Norman and MallaThere were seven studies that reported a neutral impact of strike action. These studies were again diverse conducted in the UK, Canada, India, Spain, Finland and two in the US. Strikes varied in length from 13 days in Finland to 69 days in India. Five studies examined strikes that involved doctors; two studies examined strikes that involved ambulance workers and healthcare staff. The outcomes examined included paediatric pneumonia, high‐risk deliveries, nosocomial infection, appendectomies. Three studies examined more general mental and general health outcomes or multiple outcomes.Looking more closely at these studies, Crocker et al.3.5One study reported a positive impact of strike action. Sigal et al.4This paper sought to synthesise and analyse the empirical literature on the impact of strike action on patient morbidity in an effort to understand if strike action has an impact on morbidity and if so what factors related to the strike or patients impacted these outcomes. As a whole, the literature suggests that strike action has little impact on patient morbidity. The majority of studies reported that strike action had a neutral or mixed impact of strike action on patient morbidity. One study reported positive outcomes and three studies reported negative outcomes, in each case however and with the exception of NjugunaThe studies included in this review were relatively heterogeneous in the outcomes they examined and the context in which they occurred, so there is a general need for caution in how these results are interpreted. Furthermore, we have been deliberately broad in regards to the studies included here. Some overlap substantially with the provision of services. For example, it is arguable that some studies measure disruption to service rather than patient outcomes. For example, while NjugunaOver the last few years and since the COVID‐19 was declared a global pandemic strike action appears to have become increasingly common across the globe.The authors declare that they have no competing interests.Ethics approval was not sought or required for this study.


The design and manufacturing technology of interference-absorbing short-wave filters based on a layered composition of Si–SiO on a sapphire substrate of various shapes was developed. A transition layer of SiO was applied to the surface of the substrate, alternating with layers of Si–SiO with an odd number of quarter-wave layers of materials with high (Si) and low refractive indices (SiO), and the application of an outer layer of SiO as an appropriate control of the materials’ thickness. The optical properties of the infrared light filter were studied. It was established that the created design of the light filter provides the minimum light transmission in the visible region of the spectrum from 0.38 to 0.78 µm and the maximum in the near infrared region from 1.25 to 5 µm and has stable optical indicators. A method for studying the stress–strain state and strength of a multilayer coating of a light filter under the action of a local arbitrarily oriented load was developed. For simplicity in the analysis and for obtaining results in the analytical form, the one-dimensional model of the configuration “multilayer covering—firm substrate” constructed earlier by authors was used. From a mechanical point of view, the upper protective layer of the multilayer coating was modeled by a flexible plate, and the inner operational composite N-layer was subjected to Winkler’s hypothesis about the proportionality of stresses and elastic displacements. Devices equipped with various designs of optical filters  for optiAccording to the ISO 20473:2007 standard, infrared radiation is divided into three ranges: near infrared radiation—from 0.780 to 3 μm; mid-infrared radiation—from 3 to 50 microns; far infrared radiation—from 50 to 1000 microns.As optical elements for the near-infrared region of the spectrum, multilayer interference-absorption short-wave cut-off filters are used as part of powerful optical devices (exit window). These filters adjust the spectrum of the broadband emitter, passing infrared radiation and absorbing/reflecting visible radiation. It is known that the reflection coefficient of the light filter increases with the increase in the number of layers, and an odd number of layers exerts a greater influence on the reflective capacity of the interference coating than an even number.The Doublet Metalens  was deveThe design of an ultra-broadband and highly efficient beam splitter based on a quasi-continuous metasurface in the near-infrared range, proposed in , consist3) and yttrium oxide (Y2O3) dielectric, with a critical defect of the high-temperature superconductor, yttrium-barium-copper oxide. It is a superconductor-based multichannel photonic crystal optical filter tunable in visible and telecommunication windows at cryogenic temperature. In [The authors  developeture. In , an inte2O5) and based on alternating layers with a high and low refractive index, respectively. The researchers [The study  proposedearchers  developeWhen designing filters, researchers mainly take into account recommendations ,12. They2 oxide films with Sn improves their photocatalytic properties, and carbon with palladium nanoparticles provides it with electrocatalytic properties [Computer modelling is widely used to predict photovoltaic properties , phase coperties ,26.During the construction of light filters at the stage of design and technological preparation of production, it is advisable to take into account technological heredity  to ensurResearchers  proposedWear-resistant coatings are formed by thermovacuum evaporation on piezoceramic materials and silicon probes for atomic force microscopy ,36. To iOne of the modern trends in the development of methods for assessing the strength and durability of modern structures is the improvement of interference-optical methods of the mechanics of a deformable solid body. The essence of these methods is that transparent materials become optically anisotropic under the influence of mechanical stresses . The useCalculation of thin-walled layered shells under radial load str given in works ,48, and There are well-known methods for solving problems of thermal conductivity ,55,56 anAs the above review shows, researchers have not paid due attention to the study of the optical and physico-mechanical properties of layered coatings under extreme conditions of high temperatures, under the action of arbitrarily oriented local mechanical loads, and the defectiveness of their structure.This study aims to investigate the optical properties and develop an engineering method for calculating the stress state of the layered coating of the interference-absorption filter under an arbitrarily oriented local mechanical load.producing experimental samples of interference light filters of various configurations;investigating their optical properties at different lengths of the light flux;developing an engineering methodology for calculating the stress state of the layered coating of the interference-absorption filter under an arbitrarily oriented local mechanical load and to investigate the stress state of the composition.To achieve the goal, the following tasks should be solved:First, the workpiece of the sapphire substrate was cut and mechanical processing was carried out—diamond grinding and polishing. After that, washing, degreasing, and drying were carried out. The substrate blank prepared in this way was installed in the chamber of the device for forming light filter layers, the chamber was evacuated, and the sapphire substrate was preheated. On the flat surface of the substrate, a transition layer of SiO was deposited (by resistive method) and its thickness was controlled; a layer of Si was deposited (by electron beam method sputtered) and its thickness was controlled; a layer of SiO was then deposited and its thickness was controlled; and again a layer of Si was deposited and its thickness was controlled, etc., until the given was an odd number of light filter layers  has a high value of thermal conductivity, while meeting one of the important requirements—effective heat removal under high temperature loads during use in powerful infrared optics. High-resistance silicon and silicon monoxide fraction 2 were used for sputtering the layers. All the materials used were particularly clean in terms of purity. Si and SiO materials have high adhesive characteristics (bond strength) and satisfactory resistance to the formation of residual thermal stresses both to the sapphire substrate and to each other, which will ensure high optical and mechanical characteristics of the wear composition and a long service life in extreme conditions.−3 Pa. The VU-1A vacuum sputtering unit was equipped with a UELI-1 electron-beam evaporation source and a resistive heater, as well as a SFKT-751 thickness photometric control complex . The SFKT-751 photometric control complex , St.-Petersburg, Russian Federation) was designed to control the optical thickness of films that form a coating during their application on VU-1A type vacuum units. The self-writing two-coordinate device N-307  was designed for registration in rectangular coordinates in linear or logarithmic scales of the functional dependence of two measured values, presented in the form of electrical signals of electric current voltage. The materials for sputtering the Si layer were placed in a crucible, and the SiO layer in a boat. The accelerating voltage was 12 kV, and the current was up to 250 mA. Silicon layers were sputtered by electron beam method, and silicon monoxide was deposited by resistive method. Control of the spraying process of coating layers was carried out experimentally by the photometric method using the SFKT-751 system and the N-307 device. The control wavelength was 690 nm. The measurement system consisted of the following main parts: a radiation source, a monochromator, a photoreceiver, an amplification unit, and a registration unit. The device uses the phenomenon of interference: under monochromatic illumination, the intensity of light falling on the sample increases (decreases) to ¼ the optical thickness of the film, and then begins to decrease (increase) to ½ the optical thickness. Thus, the thickness of the film can be determined by the formula: n∙d = m∙(λ/4), where n is the refractive index; d—thickness; m is the number of maxima or minima; λ is the wavelength of monochromatic light. The two-coordinate self-recording device N-307 registers extremes of light intensity.For conducting the research, samples of interference light filters of a round shape with a diameter of 30 mm and a rectangular shape with a length of 150 mm and a width of 80 mm were made with a thickness of sapphire substrate of 3 mm. The Si–SiO interference multilayer coating was applied to a specially prepared sapphire substrate preheated to 300 °C using thermal and electron beam evaporation methods in a VU-1A vacuum unit under an excess vacuum of 3·10The total number of Si–SiO layers is chosen depending on the functional purpose of the light filter. The produced interference light filter contained a transition layer with a low refractive index (SiO), an odd number of quarter-wavelength optically thick layers of materials with high (Si) and low refractive indices (SiO), the layers of which alternate, and an additional outer layer with a low refractive index (SiO), which has a larger optical thickness .To study the optical properties of light filters, optical transmittance studies were conducted for the ultraviolet, visible, and infrared regions of the spectrum, using round and rectangular samples.-Optizen 3220 UV  double-beam spectrophotometer, which is designed for measuring transmission coefficients, optical density and scanning transmission or absorption spectra in a given wavelength range of ultraviolet and visible radiation from 190 nm to 1100 nm;-Spectrometers Spectrum One FT-IR  in the wavelength range from 1.25 μm to 10 μm.Optical transmission tests of light filter samples were performed on:Measurements were performed with a scanning step of 1 nm.The structural elements of the interference-absorption filter  consist In an effort to obtain the final result in an analytical form, in this paper we developed a one-dimensional stress analysis proposed earlier for two-layer compositions ,78,79. TMechanically, we considered the upper protective SiO-layer as a tensing and bending plate interacting with the multilayered filter. At the same time, the operational compositional N-layer Si–SiO set meets Winkler’s hypothesis on the proportionality of tangential and normal stresses to respective elastic displacements. To simplify our analysis, we assumed that the sapphire substrate was absolutely rigid, and that the mechanical contact between the components on the layer interfaces was ideal. Moreover, we assumed the plane deformation state as Bd2uxLet us define the coefficients of substrate rigidity for the multi-layer depth-inhomogeneous filter as the values inversely proportional to the total compliance of series-connected layers:The forces and moments vanish at infinity:Thus, the boundary problem (1), (2) describes the required field of displacements of the coating-substrate/plate on the elastic layered substrate.The technological features of round and rectangular light filter manufacturing are very different, especially when their geometric dimensions are increased. The asymmetry of the sample shape can affect the inhomogeneity of the functional characteristics of the product’s layered coating on the plane. Therefore, samples of light filters of various shapes and sizes were produced, and their optical and mechanical characteristics were investigated. The general appearance of light filter samples with Si–SiO interference coatings formed on round and rectangular sapphire substrates, manufactured according to the developed technology is presented in The analysis of the photos  shows thA review of the pictures presented in The results of the studies of the optical transmission of light filters, conducted in different ranges of radiation wavelengths from 0.19 μm to 1.1 μm and ranges from 1.25 μm to 10 μm, are presented for round samples in As we can see from In order to study the optical properties of the large-sized light filter, we conducted a study of optical transmittance for different surface areas of rectangular samples in the ultraviolet, visible, and infrared regions of the spectrum. The measurement results are presented in The analysis of the results of the study of the transmission capacity of light filters for the ultraviolet, visible, and infrared regions of the spectrum  shows thSince the developed light filters are proposed for use in optical pyrometers, where the entire surface is used during operation, the uniformity of optical transmission over the entire area of the sample will affect the accuracy of measuring the temperature of objects. The uniformity of the thickness of the applied coating layers on the surface of the large-sized light filter samples is ensured by the use of special equipment installed inside the chamber of the vacuum-spraying installation.The analytical solution to the boundary-value problem (1), (2) is built in the following form:The corresponding force and bending moment in the coating are obtained using Equation (3) for the displacements:The stresses in the coating are linearly distributed across the thicknessIn particular at the bottom base of the coating –(8) were analyzed for the composition containing the silicon oxide coating with the parameters: As we can see on represented graphs, the most dangerous place is the point The admissible loadings Taking into account that the theoretical strength of The design of an interference-absorbing short-wave filter based on a layered composition of Si–SiO on a sapphire substrate has been developed. The light filter manufacturing technology includes the application of a transition layer of SiO to the surface of the substrate, alternating Si–SiO layers , and low refractive indices (SiO), alternating layers), completing the process of forming the light filter coating with the application of an outer SiO layer with appropriate control of the layers’ thickness.Based on the results of the optical properties study, it was established that the light filter provides minimum light transmission in the visible part of the spectrum from 0.38 to 0.78 µm and maximum light transmission in the near-infrared region from 1.25 to 5 µm and has stable optical indicators over the entire surface area of the samples.The main feature of the proposed technique of mechanical analysis is the use of strength criteria for all components of a partially homogeneous layered structure. For the layered Si–SiO composition on a sapphire substrate, the stress–strain state and the allowable load were evaluated and their dependence on the angle of inclination of the applied load was analyzed. The calculation method proposed by the authors will allow engineers to control, in the analytical form, the influence of the ratio of geometric and mechanical characteristics of nanolayers on the stress state and the limit equilibrium of the interference-absorbing filter depending on the magnitude and orientation of the localized load.Studies have shown that the most dangerous stresses are in the coating, which should be taken in account in the strength calculation, and the worst force inclination angle is the case of perpendicular loading.In the future, the effect of heating on the change in the stress–strain state of the layered light filter will be investigated.


Left side traumatic diaphragmatic hernias (DH) are very rare and usually present acutely. They might represent after years of minor trauma, and they should be considered among differentials to avoid complications. We present a 28-year-old female coming with acute epigastric pain radiating into the chest with dyspnea and vomiting. Her history was negative for trauma and other than very minor trauma two years earlier. Chest X-ray showed atelectasis with mild pleural effusion. Computed tomography scan showed several cavities, filling the left chest with a gaseous liquid level. Surgery was performed that demonstrated DH and the abdominal viscera were returned to the abdomen without any complications. Traumatic DHs can be easily overlooked with the absent of recent major trauma. They can represent years after the original trauma with acute symptoms, which can make it hard to diagnosis if not considered. Traumatic injury to the diaphragm is relatively uncommon, with an incidence between 0.8 and 8% . We presA 28-year-old housewife who presented with dyspnea and vomiting for one week, with epigastric pain radiating into the chest. She had no fever, cough or hemoptysis. She had no significant history for cardiac or pulmonary disease, and she was a non-smoker. She did not take any medication, including oral contraceptive pills. No significant family history other than diabetes and high blood pressure was found. The patient recalled carrying heavy weights around a year ago without a previous trauma story, and she had a subsequent dyspnea with a compressive intermittent chest pain that increased after eating at that stage, which all resolved spontaneously after a few days. She had a history of three Caesarean sections, with the last operation being four years ago.2. Other systems were also normal.On examination, vitals were within normal range. However, high-pitched sounds were noticed in the left hemithorax. Cardiovascular and abdominal examination were otherwise remarkable. Her BMI was 23.4 kg/m3 = 20). Chest x-ray (CXR) of the chest was not able to visualize the left hemidiaphragm with a hollow viscus in the left thoracic cavity and a right shift of the mediastinum  and respiratory alkalosis  . The claDiaphragm defects are not easily detected because there are often no direct signs and symptoms. Moreover, most accompanying symptoms are nonspecific .High clinical suspicion is required to make the diagnosis of diaphragmatic rupture; CXR is usually the first hint regarding the possibility of a diaphragm injury , 6. In tIn this case, when the woman was presented, there was a large hernia with much protruding viscera, which led to a deterioration in the general condition, and thus a significant increase in the risk due to the size of the hernia being 3 cm. Therefore, the patient was rapidly managed due to the concerns of developing incarcerated hernia where mortality rates are very high . Such prCXR might give a misleading diagnosis as it might resemble a hydatid cyst or pneumothorax, which might lead to mismanagement such as inserting a chest tube, which might cause a perforation in the stomach and colon; therefore, great care should be taken when dealing with other similar cases .Traumatic diaphragmatic hernias are rare and can be caused by relatively minor trauma that might be direct or indirect. This can occur even years after the initial trauma. Early diagnosis and management of symptomatic causes is crucial to avoid serious complications.


HPV was detected in nearly all (82%) samples, , and nearly all HPV-positive samples included types in multiple genera (88%). A total of 560 HPV detections  were made. The most frequently detected HPV types were alpha , beta , and gamma . High-risk alpha types  were not common. A novel gamma type was identified  along with 90 unclassified types. This pilot study demonstrates the utility of the eWGS assay for broad-spectrum type detection and suggests a significantly higher type diversity in males compared to females that warrants further study.Most human papillomavirus (HPV) surveillance studies target 30–50 of the more than 200 known types. We applied our recently described enriched whole-genome sequencing (eWGS) assay to demonstrate the impact of detecting all known and novel HPV types in male genital samples ( Papillomaviridae that includes 49 genera. More than 200 HPV types that infect cutaneous and mucosal surfaces have been found in five of these genera, with most in alpha, beta, and gamma genera. While most infections clear, persistent infection with some alpha types  is a risk factor for anogenital and oropharyngeal cancers [Human papillomaviruses (HPV) are DNA viruses in the family  cancers , the ass cancers . Individ cancers . Gamma HCurrently, most studies have used PCR assays with degenerate primers  to detect and type HPV within specific genera ,13. Epidn = 50) from external genital swabs in specimen transport medium collected from US males between 2010 and 2011 (IRB determination—Exempt). The vaccination history was unknown, but collection was prior to the US recommendation of routine HPV vaccination for males. DNA extracts were prepared using a Chemagic gDNA kit with an automated Chemagic MSM1 extractor  and stored at −80 °C for 63–68 months prior to use in the eWGS assay. The DNA concentration of the thawed extracts was determined using a Qubit dsDNA HS assay .We used residual anonymized DNA extracts , HPV-negative placental DNA was added to bring the amount of DNA/sample to 100 ng prior to shearing . Librarihttps://pave.niaid.nih.gov/) (as of 24 February 2022). A sample was considered positive for an HPV type if there were at least 100 mapped reads that covered at least 20% of a classified or unclassified reference HPV genome. This cut off is more stringent than recommended (minimum 10 reads and 10% genome coverage) for determining positive signal in HPV whole-genome studies [The sequence data were analyzed using CLC Genomics Workbench . Read de-multiplexing, quality control, alignment, and HPV type determination were performed as described previously , except  studies .2 and 995 K/mm2, with a total of 30, 935, and 33,083 mega base (Mb) sequence data, respectively. All samples with DNA generated an average of 10.5 million reads/sample with a mean Q score of 36.2; 93.4% of the bases had a Q score ≥ 30. Results from analysis of these data are presented below. The 13 control samples gave the expected results (Sequencing libraries 1 (samples 1–31) and 2 (samples 32–63) generated similar cluster densities of 962 K/mm results . Of 50 male genital samples, 41 (82%) were HPV eWGS-positive. There were 560 total HPV detections of two-hundred thirty-seven unique HPV types in four genera: forty-six alpha, fifty-five beta, one-hundred thirty-five gamma, and one mu types. Within genera, the types were spread across forty-two species: fourteen alpha, five beta, and twenty-three gamma species. The number of HPV types per sample ranged from 1 to 85 (mean = 13.6 types per sample). Of the 560 HPVs detected, 72% (402) were officially classified and 28% (158) were unclassified in the PAVE database. Gamma HPV detections dominated , followed by beta , alpha , and mu genomes . Among the 122 alpha HPV detections, 46 unique types were identified in 32 samples. The number of reads/alpha genome was 88,996 ± 29,619 and >80% of the genome was sequenced in over 75% (94/122) of these detections. Among the 179 beta HPV detections, 55 unique beta types were identified in 35 samples. The number of reads/beta genome was 17,020 ± 4769, and >80% of the genome was sequenced in over 57% (102/179) of the detections. Among the 257 gamma HPV detections, 135 unique types were identified in 36 samples. The number of reads/gamma genome was 36,585 ± 9738, and >80% of the genome was sequenced in nearly 42% (107/257) of the detections. The two mu papillomavirus signals were mapped from two samples  to the same unclassified novel type, HPV-md01c06. 2 = 1) with the depth of coverage (p = 0.0057) and gamma genomes . The GC content (%) was significantly higher in alpha genomes  compared to both beta  and gamma genomes   .The number of alpha types detected per sample ranged from 1 to 21 (mean = 4/sample). While the most common alpha types were HPV90, HPV43, HPV74, and HPV84 A, the grThe number of beta types detected per sample ranged from 1 to 26 (mean = 5/sample). While the most common beta types were HPV types 115, 195, 120, 22, 23, 24, and 120 A, the grThe number of gamma HPV types detected per sample ranged from 1 to 53 (mean = 7/sample). While the most common gamma types were HPV134, HPVmSD2, HPV 50, HPV179, HPVmKN1, and HPV147 A,B, the The overall prevalence of male genital HPVs belonging to alpha, beta, and gamma genera is presented in Interestingly, of the 237 unique HPV types identified by eWGS, there were no RNA baits in the enrichment hybridization for ninety-seven types  and eighty-three of these types were unclassified. The study also identified a novel HPV genome (GenBank Accession # MW535770), that has subsequently been officially classified by the International HPV Reference Center, Sweden, as gamma HPV type 229.Papillomaviridae genera recognized to have HPV types   . For thrIt is interesting to note that of the five most prevalent high-risk types in a meta-analysis of data from women with normal cervical cytology in the pre-vaccine era  , HPV 58 Alpha types, the major genus detected in cervicovaginal samples, were not the most frequently detected in male genital samples, although they were present at a higher copy number (averaging two to five times more reads/sample than beta and gamma genomes) and had the highest percentage  of complete genomes. It is also striking that the HPV diversity in male genital samples in our study was greater than that reported in cervical samples tested with the same eWGS assay ,25. PrioPrior studies of male genital samples using multiple assays (linear array followed by sequencing of PCR products of PGMY 09/11 and FAP59/64 primer systems) found fewer than 20 unique beta types largely in one beta species (beta 2) . HoweverWe identified 55 of the 98 types within gamma, the genus with the largest number of types with official classification. Of the unclassified types detected in our study, the largest number  were in this genus. Among gamma types, the prevalence was highest for HPV-mSD2 and HPV 134, each at 14%, followed by HPV-mKN1, HPV 50, and HPV147, each at 12% in this study. HPV-mSD2 and HPV134 were previously identified as the most prevalent gamma types in oral rinse samples . This pilot study has several limitations. It used residual samples without demographic information, and results cannot be considered representative of any population. Storage may have compromised sample quality, although quality control suggested this was not an issue. HPV vaccination status was unknown, although the dates of the original sample collection were before widespread male vaccination. Enrichment methods, particularly those that rely on PCR, may bias the observed type distribution. As eWGS uses a limited number of PCR cycles and maintains a strong linear relationship between target copies and signal strength , this coThis pilot study demonstrates the usefulness of eWGS for broad-spectrum detection of HPV genomes belonging to multiple genera. Systematic studies using eWGS will generate a more complete view of HPV type distribution, to better understand their biology, tissue tropism, and potentially new disease associations.