Ovarian cancer is the most lethal malignancy among gynecological cancers worldwide. Most ovarian cancer patients are diagnosed at an advanced stage because of non-specific clinical symptoms. The human microbiome plays a crucial role in maintaining the normal physiological and pathological state of the body. With the development of technologies such as DNA and 16S rRNA sequencing, an increasing number of findings on the role of microbiome in cancers are being reported. Microbiome abnormalities are increasingly associated with diseases, including cancer development, and response to therapies. Some studies have shown the relationship between microbiome changes and ovarian cancer. However, the mechanisms underlying this relationship are not yet fully understood. Here, we summarize the key findings in this regard by focusing on estrogen metabolism and host recognition receptors in microorganisms and changes in the gut or pelvic microbiome in patients with ovarian cancer. We further discuss the potential of using the microbiome as a novel biomarker for cancers. We also highlight the possibility to use microorganisms as a treatment modality to enhance the immune system, activate anti-tumor response, mediate chemotherapy resistance, and ameliorate the adverse effects of the treatment. Ovarian cancer is the most lethal gynecological malignancy, with more than 200,000 new cases and 150,000 deaths reported annually worldwide . The falThe human microbiome is a complex ecosystem housing a variety of microorganisms residing mainly in the epidermal and mucosal tissues of the human body . The terThe entire human genome comprises ~25,000 human genes, whereas the resident microorganisms contribute more than 1 million genes . The humThe intestinal bacteria are of three types: beneficial, harmful, and neutral. The intestinal or gut microbiome is highly diverse and plays a role in synthesizing many essential vitamins and amino acids while promoting the digestion and absorption of various minerals. Moreover, the gut microbiome plays an important role in maintaining an autoimmune barrier and enhances the mucosal immune system. Thus, intestinal dysbiosis can affect the nervous and immune systems by interacting with the Vagus nerve and by destroying the integrity of the epithelium, respectively . AbnormaHelicobacter pylori, fusobacteria, and other potentially infectious pathogens may also increase the risk of colorectal cancer  than women with the lowest estradiol levels . In addiOne study investigated the relationship of the diversity and composition of the fecal microbiome with urinary estrogens and estrogen metabolites and found that postmenopausal women with a more diverse gut microbiome had a higher proportion of hydroxylated estrogen metabolites in their urine . This fiMicroorganisms mainly interact with human cells through the combination of microbe-associated molecular patterns on the microbial surface and corresponding receptors of the host . These iNuclear factor-kappa B (NF-κB) is the downstream target of TLR, which can stimulate the release of inflammatory cytokines. A hallmark of innate immunity is the proper recognition of potential pathogens and harmless antigens through PRRs. Activation of PRRs has been suggested as a cause of chronic inflammation and carcinogenesis . For exaNLRs are the innate immune PRRs that can activate multiple pro-inflammatory responses and lead to antigen-specific acquired immunity via downstream signaling. NLRs are involved in the occurrence, development, and pathology of cancers such as glioblastoma, prostate adenocarcinoma, breast cancer, melanoma, and colon adenocarcinoma . NLRP3 iCLRs, which harbor at least one C-type lectin-like domain, play an important role in antifungal and antibacterial immune responses, regulation of immune homeostasis, autoimmunity, allergy, and recognition of dead and cancerous cells . CLRs paIn addition to treatment resistance, late-stage diagnosis is a primary reason behind high mortality rate of patients with ovarian cancer. The disease is often diagnosed late mainly because of the lack of specific symptoms. One of the main hallmarks of cancer is the chronic inflammatory state. High-grade serous carcinoma (HGSC) with a high mortality rate is the most common subtype of ovarian cancer. Preventative salpingectomy in high-risk populations could prevent HGSC as well as endometrioid and clear cell carcinoma. It inhibits the spread of potentially cancerous cells into the ovarian/peritoneal surfaces and the ascension of endometrium and related inflammatory factors . HoweverChlamydia trachomatis and Mycoplasma genitalium are sexually transmitted microorganisms associated with pelvic inflammatory disease. The presence of chlamydial HSP60-1 IgG antibodies is associated with high-grade ovarian epithelial carcinoma, while the presence of M. genitalium IgG antibodies is associated with borderline ovarian tumors  in mice and patients could also favor the efficacy of CTLA-4 blockade , and immune checkpoint inhibitors, have shown good performance in delaying recurrence and prolonging the overall survival time of patients. For example, ovarian cancer patients with BRCA1/2 mutations treated with PARPi therapy showed an ~4-fold longer progression-free survival compared with those taking the placebo, whereas the increase for patients without the BRCA1/2 mutation was ~2-fold . HoweverCisplatin is used as a first-line treatment for advance-stage ovarian cancer. Intestinal damage is an adverse effect associated with cisplatin, which results in modifications in the process of treatment, such as dose reduction and delaying or cessation of the treatment . AlfredoIn short, microbiome can change anti-tumor activity, mediate chemotherapy resistance, and reduce the number and intensity of adverse effects in the process of cancer treatment.The microbiome plays a crucial role in maintaining the health of the human body, and abnormalities in the microbiome have been associated with a variety of diseases, including ovarian cancer. With the development of technologies such as high-throughput DNA and 16S rRNA sequencing, more and more aspects of the human microbiome are being revealed. Probiotic bacteria show the potential to become a significant component in the prevention and treatment of cancers . DespiteHC drafted the manuscript. HC and ZW were responsible for the conception, planning and carrying out the study, reviewing the literature. YW, LC, XC, and FG were responsible for discussing and final review of this manuscript. HY was responsible for the conception and final review of this manuscript. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


African swine fever (ASF) is a highly lethal contagious disease of swine caused by African swine fever virus (ASFV). At present, it is listed as a notifiable disease reported to the World Organization for Animal Health (OIE) and a class one animal disease ruled by Chinese government. ASF has brought significant economic losses to the pig industry since its outbreak in China in August 2018. In this review, we recapitulated the epidemic situation of ASF in China as of July 2020 and analyzed the influencing factors during its transmission. Since the situation facing the prevention, control, and eradication of ASF in China is not optimistic, safe and effective vaccines are urgently needed. In light of the continuous development of ASF vaccines in the world, the current scenarios and evolving trends of ASF vaccines are emphatically analyzed in the latter part of the review. The latest research outcomes showed that attempts on ASF gene-deleted vaccines and virus-vectored vaccines have proven to provide complete homologous protection with promising efficacy. Moreover, gaps and future research directions of ASF vaccine are also discussed. Sus scrofa) and domestic swine of all breeds and ages. ASF with the acute form has a rapid onset and a short course of disease, which are characterized by high fever, loss of appetite, cyanosis, severe bleeding of internal organs and a high mortality rate of nearly 100% [African swine fever (ASF) is a highly contagious animal infectious disease caused by African swine fever virus (ASFV) infecting both wild  [ASFV is the sole member of the es (MGF) , and ences (MGF) . Morpholes (MGF) . ASFV reThe genome of ASFV is huge and complex, showing obvious genetic diversity. There are currently eight serotypes based on the viral hemagglutinin CD2-like protein (CD2v) and C-type lectin. According to the sequence of B646L, which encoding the major capsid protein p72, ASFV can be divided into 24 genotypes (I-XXIV) . ASFV waRecently, the World Organization for Animal Health (OIE) issued a report on the “Global Situation of ASF” which indicated that there has been a marked pattern of increasing ASF outbreaks worldwide since 2016. As of 18 June 2020, 30% (60/201) of the reporting countries/regions had reported the presence of the disease, Since 2020, 8330 ASF outbreaks have occurred in 25 countries and regions worldwide, including 1604 swine and 6726 wild boars, showing a serious deterioration in the incidence of the disease, especially in Europe and Asia. Europe accounted for 67% of the outbreaks reported through immediate notifications and follow-up reports, notably, Asia  accounted for the largest proportion of reported pig losses. OIE encourages interregional cooperation to combat the spread of this cross-border transmitted disease [At present, effective control of ASF has not yet been achieved in Asia and Europe, which poses a serious threat to the global pig industry. No available vaccines against ASF leaves the control of the disease to early detection by efficient diagnosis, the culling of infected and exposed animals, and strict sanitary measures. Lack of in-depth understanding of the prevalence, transmission mechanism and pathogenicity of ASF has brought great difficulties to the prevention and treatment of ASF in China. The purpose of this review is to summarize the current epidemic situation and factors of ASF in China. The second half of this review covered the recent progress and emerging approaches to ASF vaccine design. We look forward to delivering helpful information for the next generation of ASF vaccines to leap over hurdles.ASF has brought a grievous blow to China, which is the world’s largest pork producer and consumer, with pork production accounting for about 53% of the world’s supply. Further, pork is the main source of high-quality protein for Chinese residents, with the consumption accounting for 62% of total meat consumption . Until 2ASF has been listed as a foreign animal disease with focused surveillance all along in China. After the first report of the ASF epidemic on 3 August 2018, the MARA immediately initiated an emergency response and took various preventive and control measures in time, but still failed to curb its spread. The following analyzed the influencing factors of the ASF epidemic in China.The ASFV genotype II strain prevalent in China has strong pathogenicity and infectivity, and is highly contagious and stable in the environment . The virASFV has complex and diverse transmission mode, it can circulate among domestic pigs-wild boars, wild boars-soft ticks-pigs, and can be propagated in pigs through direct and indirect contact or short distance aerosol ,25. ASFV can be transmitted horizontally in infected pigs. The virus content in the blood of highly acute attack pigs is very high, which provides favorable conditions for the direct or indirect transmission of the virus. The virus also exists in urine, nasal secretions, saliva and other excreta and spreads through short distance aerosol. In addition, direct contact with contaminated surfaces, feed feces or water can indirectly lead to pig infection. All of these pathways contribute to the persistence of ASFV in pigs . AlthougSus scrofa) lack resistance to ASFV and have a high mortality rate once infected. Since 2014, 95% of reported ASF outbreaks in Europe have been wild boar infections, and ASF persistence has been maintained through transmission between wild boars in the absence of domestic boars [2, and widely distributed in Fujian, Guangdong, Northeast China, Yunnan-Guizhou region, etc. It is estimated that the number of wild boars in the whole country has reached millions now [Wild boars have a wide scope of activities, ASFV can survive in infected wild boar carcasses and environment for months, and mechanical transmission of pathogens will occur once the carcasses are eaten by other carnivores or birds. Different from warthog and giant forest pigs in Africa, Eurasian wild boar . Line-haul of infected pigs directly results in rapid spread of the virus. Legal trade of live pigs may spread ASF during high-risk periods (HRP). Gao Xiang et al. found that the southeast of China had the highest risk of ASFV transmission through legal pig trade, especially in colder months . TherefoAccording to previous statistics, domestic pig circulation is the main route of ASF transmission in China. Infection can occur when feeds and swill contaminated with ASFV from epidemic areas are ingested in healthy pigs . Most ofIn developing countries, most pig breeding remains traditional, small-scale, and self-sufficient. Small-scale family farming plays a major role in the introduction, dissemination and colonization of ASF due to its low biosecurity, backward feeding methods and techniques, insufficient understanding of animal health regulations, or even non-compliance with regulations . At presThere is no fixed model for the prevention and control of ASF, a Chinese model for the control of ASF is being established with absorbing and drawing on the experiences of foreign countries. At present, due to the decline of pig stocks, and benefited from the improvement of biosafety awareness and biosecurity level of pig farms in large scale, ASF epidemic has been monitored and detected by a large number of breeding enterprises, and it is slowing down in China . HoweverCurrently, there is no safe, effective and commercially available ASF vaccines, so there is no way to control susceptible animals. The prevention and control of ASF in China focuses on eliminating the contagium and polluter, and cutting off the transmission route, which will be a long-term work. Contagium and polluter mainly include dead and infected pigs, especially the infected pigs without clinical symptoms, which should be culled and harmlessly treated, and forbidden to enter the transportation, sale, and slaughtering links. It is necessary to strengthen the cleaning and disinfection of transport vehicles and the isolation of personnel; drying, high temperature granulation, acidification of polluted feed materials; drying and disinfection of polluted materials, disinfection and acidification of polluted water sources, cleaning and maturation of polluted food materials, and the measures mentioned above are intended to completely inactivate ASFV in the environment. In addition, appropriate measures should be taken to prevent the mechanical transmission of ASFV by fly/mosquito/bird. Finally, transport, slaughter, and feed enterprises, meat processing enterprises should do a good job in the biosecurity system of ASF and adopt some corresponding technologies to eliminate potential pollution sources ,39.(1)Introduction control. Introduced breeding pigs must be isolated for more than 30 days and tested clinically and laboratory to ensure that ASFV was negative.(2)Transport vehicle control. Establishing a standardized cleaning and disinfection system and specifications for transport vehicles, and separate and specific trucks are recommended to transport the pig and material.(3)Control of items entering pigsties. Strict disinfection treatment of items entering pig farms by fumigation, ozone and other methods.(4)Personnel control. Personnel inside the pig farm should reduce their outgoing activities and isolation measures for admission personnel should be implement strictly.(5)Feed control. Eliminate potential sources of contamination of feed.(6)Environmental control. The surrounding environment of pig farms should be regularly monitored, detected and evaluated.New pig farms should meet the biosecurity requirements for the prevention and control of ASF, adhering to the general principles of “Scientific Design, Zonal Layout, Physical Isolation, Zonal Grading, Intelligent Management”. In order to prevent the spread of ASFV in pig farms and control the epidemic to a minimum extent and scope, we should strengthen the construction of the biosecurity system in pig farms and strictly implement the corresponding sanitary measures , which mMoreover, the OIE also encourages members to implement enhanced national sanitary measures on waste disposal from aircrafts/vessels/passengers and enhanced in-farm biosecurity measures–including the protection of pigs from untreated swill feeding and the effective separation between domestic pigs and wild boar–and stresses the importance of OIE international standards for risk management of transboundary animal diseases (TADs) to reduce the risk of exporting disease to trading partners . Although ASFV was discovered as early as the beginning of the 20th century, vaccine research has lagged behind for more than 40 years, which may be related to its obvious genetic diversity, showed by the very large and complex genome of ASFV. Thus far, the exact mechanism of the host protective response has not been fully determined and protective antigens remain to be identified, which seriously hinders the development of vaccines. The lack of effective treatments or vaccines against ASF complicates the control of ASF in China. Consequently, we reviewed the research history and recent progress of ASF vaccines in order to provide reference for the research and development of ASF vaccine in China. Despite the obstacles, a few laboratories devoted to ASF vaccine research have made several pioneering discoveries in this field. For example, some candidate strains of gene-deleted vaccines can provide complete cross-protection, and are expected to be put on the market in the near future . FurtherTM or Emulsigen® -D adjuvant, but the strain failed to provide protection and acute clinical symptoms were quickly observed [Inactivated ASF vaccines were first attempted in 1960s, but studies have shown that inactivated ASFV as a vaccine is essentially an infeasible strategy . In 2014observed . Inactivobserved .ASFV attenuated by continuous subculture can induce protective immune response against parental strains. In 1960s, the attenuated strain was widely used in Spain and Portugal, but it was abandoned due to the chronic infection symptoms of ASF, which extensively appeared in vaccinated pigs. Nevertheless, The ASFV Georgia strain, which were fully attenuated after 110 consecutive passages by Vero cells, cannot resist the attack of the parental ASFV Georgia virulence after immunizing pigs, and may not produce an effective immune response due to the change of its antigen during passage . In the ASFV encodes a variety of proteins to inhibit and evade the host immune response by regulating host cell protein expression, interfering with the innate immune system and regulating cell cycle, which creating favorable conditions for self-proliferation and proliferation . An apprIn view of the recent five years of research on gene-deleted vaccines , most of+ T cells in vitro [In our vision, the same gene deletion in different strains do not invariably have the identical outcomes, because the effect on the attenuation and protection against ASFV may be strain dependent. In 2020, Chen et al., found that unlike Georgia 2007, the Chinese HLJ/2018 virus (HLJ/18-9GL&UK-del) with a similar deletion of 9GL and UK was attenuated in pigs, but could not provide any protection against homologous virus challenge. The researchers have shown that the molecular basis of the virulence of ASFV may vary among strains, and that the biological changes caused by gene deletion in one ASFV strain may differ in other strains . In 2017in vitro . Unexpecin vitro . This inin vitro , and thein vitro . OverallAlthough most of the gene-deleted vaccines without ASFV-related virulence genes can provide complete protection, there is still a risk of virulence returning due to the residual virulence. The new investigation showed that the attenuated strain HLJ/18-6GD obtained by deletion of six genes of MGF360/505, multigene families of China HLJ/2018 strain, could protect pigs from the attack of parental virulent stains. However, its safety assessment data indicated that HLJ/18-6GD has a high risk of converting to a virulent strain. In the group’s latest study, the HLJ/-18-7GD, which further deleted the CD2v gene, was obtained to eliminate this risk, fully attenuated, and with a low risk of converting to a virulent strain, and could provide complete protection in pigs against lethal ASFV challenge. HLJ/-18-7GD is a safe and effective vaccine against ASFV  and, as At present, the main factors restricting the development of ASFV gene-deleted vaccines are still insufficient research on virulence-related genes and immune escape mechanisms of ASFV, as well as subclinical symptoms and viremia caused by vaccination. A recent study described that a method of rapid isolation and purification of the deleted strain by combining the conventional homologous recombination with fluorescence-activated cells sorting (FACS) may greatly facilitate studies on understanding ASFV gene functions and accelerate the development process of recombinant live attenuated vaccine to some extent . It is bAt present, ASF genetically engineered vaccines, such as subunit vaccines, DNA vaccines, and virus vectored vaccines, provide a targeted approach with fewer side-effects and higher safety. As is shown in + T cells in the response to ASFV [+ T cells [+ T cells and revealing the potential protective capacity of new ASFV antigens including CP312R [It was previously thought that ASFV could not induce neutralizing antibodies. Gomez-Puertas et al., found that p72, p54, and p30 proteins have neutralizing effects, p72 and p54 could inhibit virus adsorption, p72 and p30 could activate cytotoxic T lymphocyte (CTL) response, while p30 could inhibit virus internalization . Other e to ASFV . In 2012 T cells . Additiog CP312R . The conAt present, most of the protective antigens of ASFV that used in current subunit vaccines are insufficient to provide complete protection for vaccinated pigs. On the whole, with only a few ASFV proteins as antigens, it is difficult to provide effective immune protection even if neutralizing antibodies are emerged . AlthougA large number of studies have demonstrated that virus vectored vaccine can induce strong specific antibody-mediated humoral responses and IFN-γ secreting cellular responses in pigs, which can provide partial effective protection against ASFV challenge ,97,98. ILimited understanding of what constitutes a protective and pathogenic immune response to ASFV has hindered the development of virus vectored vaccines, immune hyperstimulation seems to be a key factor affecting the course of ASF, and high levels of antibodies seem to have a particularly adverse impact on clinical outcomes and protection, and whether the focus should be shifted to less immunogenic ASFV antigens remains to be further explored.+ T cells, after understanding the key role of simultaneously inducing antibody-mediated immune responses and CD8+ T cells in immune protection.Given the fact that neither subunit nor DNA vaccine vaccination can provide complete protection, researchers have proposed new vaccination strategies, including “cocktail” vaccination, and “Prime-Boost” vaccination strategies that can rapidly generate large numbers of memory CD8In the previous discussion, we found that DNA vaccines can induce broad-spectrum immune responses, and DNA-protein immunity may help to identify virus antigen-specific responses, which can provide more potential information for the development of more efficient and safer ASFV subunit vaccines. In 2019, the combination of ASFV recombinant protein  and pcDNA3.1 encoding ASFV gene  was selected to immunize pigs with ISA25 adjuvant. The results showed that pigs vaccinated with ASFV DNA+ protein could activate cell-mediated immune response, in which p72 was the main inducer of IFN-γ . In the In recent years, vaccination of pigs with complex ASFV antigen preparations was carried out in order to find an appropriate balance between antibody and cell-mediated ASFV immune response. In 2017, Lopera-Madrid et al., found that vaccination with HEK 293-purified recombinant ASFV proteins  could promote the production of ASFV-specific antibodies, but did not enhance cellular immunity. However, enhanced vaccination with Modified vaccinia virus Ankara (MVA)-vectored antigens  could complement the former and promote the production of IFN-γ by cellular immunity . On the What antigen can play a protective role and how to orient the antigen to a specific presentation pathway are the urgent problems to be solved in the development of ASFV genetically engineered vaccine at present. Combining the diversity of existing virus strains, further exploring the potential protective antigens of ASFV and the optimal immune mechanism to be triggered after vaccination, the use of multiple ASFV antigens or antigen fragments, adjuvants, and complex formulations of expression vectors are a new paradigm for the development of ASFV genetically engineered vaccines.Since the first occurrence of ASF in China in August 2018, the epidemic has spread continuously and rapidly throughout the country. The wide distribution of wild boars and soft ticks in China may make it easier to form a natural epidemic focus of ASFV. In addition, the diversified breeding patterns, high breeding density, and weak biosafety awareness of practitioners make the situation of prevention, control, and eradication of ASF in China very serious. As there is still no commercial vaccine available, the domestic strategy for ASF prevention and control relies largely on the strict biosecurity system. Developing safe and effective animal vaccines will help control and eradicate ASF from wild boars and domestic pigs in China. Based on the above analysis of the global ASF vaccine research status, the recombinant LAVs developed with the attenuated strain of ASFV virulence gene knockout has the most optimistic prospects as a short/medium-term vaccine candidate in the future. However, before practical application, comprehensive evaluation of the safety risks, including virulence re-enhancement, adverse reactions, and persistent infection of the candidate strains, is needed. In addition, it is necessary to develop a matched diagnosis technology of DIVA, the establishment of ASFV culture passage cell lines, and small animal infection models for vaccine research and production, and, most importantly, basic research on how viruses regulate host responses to infection and the role of virus-encoded proteins in evading host defenses will contribute to developing the next generation of recombinant LAVs. At the same time, ASF subunit vaccines, DNA vaccines, and live virus-vectored vaccines have great research potential with tremendous advantages in safety and DIVA diagnostic technology, but we must consider that although these vaccines can induce strong antigen-specific humoral and cellular immune responses, they can rarely provide complete immune protection against lethal ASFV strains. Undoubtedly, we need to identify more antigens with protective potential and, on this basis, explore the combinations of antigens to induce high levels of immune protection. Moreover, the delivery systems and vaccination strategies need further optimized. Specifically, we must fully understand the structure and function of the main proteins of ASFV, the mechanism of infection and immunity, the recognition of the main immunogens and vaccine targets in order to conduct a more in-depth study of genetically engineered vaccines, and comprehensively evaluate their safety and immune effects in targeted animals. Ultimately, the mechanism of natural hosts against ASFV, including wild boar and warthog, will be helpful to target key factors in the development of ASFV vaccine. In conclusion, there is still a long way to go in the development of the ASFV vaccine.The introduction of ASFV has led to a heavy loss of pig production capacity and a devastating hit to the pig industry in China, and has disrupted the original breeding structure and layout of pig raising in China, which has a significant impact on the original pig breeding mode and business model. Nevertheless, it is also an opportunity of rebirth for the pig industry. On the one hand, it improves the threshold of breeding, promotes the transformation and upgrading of the pig industry, and accelerates its process of scale, automation, and intelligence. On the other hand, it changes the concept and enhances the ability of epidemic prevention and control, improves the level of biosecurity, rebuilds the epidemic prevention team, and remodels the epidemic prevention system and the modern pig industry.At the present stage, China has initiated special studies on major basic scientific issues of ASFV, vaccine creation, detection technology, epidemiology, special disinfectants, and vector control, which provide important scientific and technological support for ASF prevention and control in China. In addition, the phased results of ASF vaccine research and development in China have greatly encouraged the confidence to overcome ASF. As stated in the report on the situation of ASF from the World Organization for Animal Health (OIE), interregional cooperation should be encouraged to combat the spread of this cross-border transmitted disease. Controlling ASF will be a challenging long-term battle that requires the joint participation and coordination of all national and international stakeholders.


Lichen planus is an idiopathic lichenoid dermatosis that has many variants, such as the follicular form, which is clinically and histologically characterized by follicle involvement. Usually the most typical dermoscopic feature of lichen planus is the presence of whitish lines, also called Wickham striae (WS), which histologically correlate with orthokeratosis above the zones of wedge-shaped hypergranulosis . WS are Other features that can be observed at dermoscopic analysis are the presence of gray-blue dots, peppering, red lines due to the presence of vascular structures, comedo, milium-like cysts, and blue-white veils in the center .A dark-skinned man of African origin, aged 33, who has been living in Italy for the past 10 years, came to our attention after a 1-year history of a pigmented lesion on his face. This lesion was previously treated as actinic keratosis with diclofenac sodium 3% gel, but this treatment had not been successful. On examination, the area of concern showed irregularly flat, dark brown macules with defined limits on nasolabial folds. The lesion showed 2 slightly elevated darker infiltrated and palpable areas, 1 on the left side of the upper lip and 1 under the nostril . The patDermoscopy showed the typical pseudonetwork pattern with gray-brown perifollicular pigmentation, reticulated hyperpigmentation and 2 areas of darker irregular pigmentation, and obliteration of follicular openings .According to the dermoscopic features, the recent onset of the lesion, and its increased size, two 4-mm punch biopsies were taken for histopathological examination under the suspected diagnosis of atypical melanocytic lesion.The final clinical and histopathological diagnosis  was follThe particular site of the face and the dark skin of our patient can hide some typical features of follicular lichen planus and can mimic some typical features of melanocytic lesions. The pathognomonic WS may not be visible in pigmented skin and WS may not be appreciated on lichen planus lesions when the patient has been receiving topical treatment such as topical steroids or salicylic acid.Dermoscopic analysis of our case showed a gray pseudonetwork with asymmetrical pigmented follicular openings. This pattern, however, is common to numerous pigmented lesions, both melanocytic and not melanocytic lesions, which often makes a correct diagnosis difficult.Moreover, the presence of a milky pink erythema in a part of the lesion and the obliteration of some follicular openings can mean a progression of malignant lentigo .Our case shows all typical dermoscopic features of a melanocytic lesion on the face. Therefore, the preoperative clinical diagnosis was of an atypical melanocytic lesion.The presence within the lesion of a dermal nevus made the dermoscopic diagnosis even more difficult. In fact, the overall dermoscopic appearance of our lesion revealed the 4 most important features of malignant lentigo in progression: asymmetric pigmented follicular openings, dark rhomboidal structures, slate-gray globules, and slate-gray dots.This case shows the difficult clinical and dermoscopic differential diagnosis of pigmented lesions on the face, mostly with darker skins, and shows that it is important to consider also lichenoid dermatitis as a possible differential diagnosis in the management of pigmented lesions of the face.


The swine industry has an important interest in becoming more efficient and innovative, fitting with the consumer demand in terms of pork quality, meat pig welfare conditions, and the environmental aspects. This paper deals with the productive  and technological  factors that affect fresh pork and elaborates the quality of products by using proteomic tools for assessing. These technologies are a relevant approach in the meat science field to decipher the underlying mechanisms and post-mortem changes in the muscle and biofluids proteome of pigs because their study will allow better management of the outcomes such as meat quality variation and defects. In general, these new developments in molecular techniques can help researchers to control and assess this quality through biomarkers. Additionally, as food safety and pork product authentication/adulteration to avoid fraud can be evaluated with these high-throughput proteomic tools, hence they were tackled. Finally, another relevant point addressed focused on the search of bioactive peptides with a beneficial effect on human health from added-value products such as dry-cured ham. Overall, this review describes the current and emerging proteomics studies dealing with raw pork and pork products from the farm to fork.The quality assurance of pork meat and products includes the study of factors prior to slaughter such as handling practices, diet and castration, and others during the post-mortem period such as aging, storage, and cooking. The development over the last two decades of high-throughput techniques such as proteomics offer great opportunities to examine the molecular mechanisms and study a priori the proteins in the living pigs and main post-mortem changes and post-translational modifications during the conversion of the muscle into the meat. When the most traditional crossbreeding and rearing strategies to improve pork quality were assessed, the main findings indicate that metabolic pathways early post-mortem were affected. Among the factors, it is well documented that pre-slaughter stress provokes substantial changes in the pork proteome that led to defective meat, and consequently, novel protein biomarkers should be identified and validated. Additionally, modifications in pork proteins had a strong effect on the sensory attributes due to the impact of processing, either physical or chemical. Maillard compounds and protein oxidation should be monitored in order to control proteolysis and volatile compounds. Beyond this, the search of bioactive peptides is becoming a paramount goal of the food and nutraceutical industry. In this regard, peptidomics is a major tool to identify and quantify these peptides with beneficial effects for human health. The global demand and industrial production of pork and pork products are increasing. According to the Food and Agriculture Organization of the United Nations (FAO) statistics, worldwide swine production has increased steadily from 1961 to 2007, with Asia the continent with the highest production with around 50% of the total worldwide production. Between 2007 and 2017, production increased significantly in Asia and remained as the highest swine producers , since China assumed most of the production with more than 3700 million pigs produced during this time. Europe, with more than 25% of world production, is the second continent with the highest production. Germany, Poland, and Spain are the major countries with a remarkable rate in swine production [From the above, it seems that the swine industry has significant interest in becoming more efficient and innovatively fitting with the consumer demand. Consumers are now demanding higher standards for the welfare of pigs and pork quality. Factors affecting the production system , pre-slaughter conditions , and post-mortem interventions  have an impact on pork eating quality, therefore, these factors should be carefully monitored during the whole continuum of farm-to-fork, especially at the industrial level. The relationship between animal welfare and pork quality has been detailed and rigorously researched from a proteomic point of view . During One of the main problems associated with poor pork quality is the PSE  meat defect. This defect can be, for example, exacerbated by inadequate mixing with animals from different pens or batches raising fights and aggressions, increasing their stress pre-slaughter, hence causing PSE meat . StresseIn the framework of high-throughput omics technologies, proteomics offers insights about the complex network of proteins and pathways underlying variation in pork quality. The study of the post-mortem muscle proteome together with protein–protein interactions, and post-translational modifications of proteins become a challenging task to deliver high-quality meat products . In partTherefore, this review aimed to provide a summary of the current pork proteomics studies with the aim for a better understanding of the protein changes and their relationships with quality traits and the processing parameters of pork and its associated products. Thus, current technological advances and their integration with other traditional techniques are further briefly discussed. This review includes a current collection and non-exhaustive list of papers in the field, which have been discussed.For consumers, the perception of muscle color, texture, and marbling are the main aspects that drive their choice. The consumer’s first impression is strongly associated with the pork color at the point of sale whereas muscle firmness/texture and marbling are used to predict the final eating quality. This complex network of sensory attributes defined by color, pH, texture, water holding capacity, and marbling can be, for example, impacted by breed selection and rearing practices . In geneFrom a proteomic point of view, pig breed-specific differences among genotypes have shown different growth performances and fat accumulation as well as variations in the anaerobic activity of pig muscles after slaughter. These facts allow for explaining how differences in post-mortem temperature among breeds provoke differences in the activity of calpains and cathepsins, resulting in tender or tough meat . Novel aAnother relevant aspect of pig production is the residual feed intake (RFI) used to calculate the feed efficiency of growing animals at the farm. This trait may also be predicted using a proteomic approach as RFI can be correlated with molecular differences in skeletal muscle such as composition and muscle characteristics or other protein fluids such as plasma ,31. In tOn the other hand, within the swine industry, a widespread practice employed is the castration of male piglets. The main reasons why the pigs are castrated are an improvement in pig management, growth, carcass qualities, and mainly to avoid the unpleasant aroma and taste of pork perception related to “boar taint”. This latter reason is probably the most important due to the odor and flavor characteristics in raw and cooked pork meat, which causes rejection. However, the controversy about the surgical castration of pigs has led to the search for alternatives such as immunocastration. This option consists of vaccinating against gonadotropin-releasing factor (GnRH). However, there is little research about the effect of immunocastration on meat quality . Using t2+/calmodulin-dependent protein kinase II was a key enzyme with an interplay role in these processes [The relationship between pre-slaughter stress and meat quality have been demonstrated, but several unknowns and more knowledge are needed at the molecular level, as reviewed by Mouzo et al. . Apart frocesses . Additiorocesses . More sprocesses ,48. Protrocesses . Furtherrocesses . For theMeat quality is generally determined by farm production factors as well as by post-mortem interventions techniques applied at the industrial scale. Recent studies regarding the effect of different post-mortem traits aiming to assess and improve pork quality are shown in 2+, lower contents of type II fibers and those of glycogen, which decreased the extent of glycogenolysis in high-quality meats from the longissimus dorsi muscle [In the case of pork quality, proteomic technologies have been demonstrated to be very effective in the prediction of drip loss and ultimate pH from 50% to 80% of protein biomarkers in the total proteome . This tri muscle . Considei muscle . Howeveri muscle . On the i muscle . A strati muscle . It is wi muscle . Myosin-i muscle . The higIn addition to the above-mentioned, other objective of the pork industry is food authentication and the detection of adulterations. A non-extensive summary of pork product authentication/adulteration is shown in Nowadays, pork is an important part of the diet of many cultures because of its great versatility and thee abundant foods that can be manufactured from sausage to a dry-cured ham. Indeed, in recent years, most pork meat is sold as ham, bacon, and sausages than fresh pork. Further processing of pork should be considered in great detail to achieve high quality and palatability of products. The quality is associated, as evidenced above, with protein structure and lipid and protein oxidative reactions occurring during industrial processing and storage. Therefore, proteomics emerges as a relevant field, giving rise to new knowledge and understanding of the mechanisms. Regarding the functional properties of proteins, sausages contain a major insoluble protein fraction with a high content of myofibrillar proteins compared to cooked hams. This insoluble protein fraction is rich in tropomyosin and myosin light chains forming aggregates with actin. Moreover, a high level of protein oxidation was identified due to oxidized methionine in cooked products. The myofibrillar network, together with coagulated sarcoplasmic protein matrices, establishes the gel properties of cooked pork products ,73. SaltTechnical improvements were analyzed from a proteomic point of view, providing an insight into the molecular modifications occurring in the meat matrix. Regarding healthy pork products, reducing salt content without changing the sensory and textural properties is the main goal in the production of dry-cured ham. For instance, when a pressure treatment before salting was investigated, glycolytic enzymes as well as myofibrillar proteins were found differentially abundant in exudates, suggesting a faster loosening of the myofibrillar structure . Other sBeyond this technological aspect, the bioavailability and bioaccessibility of food proteins are key factors of meat products to assess their nutritional and healthiness potential . These fIn the case of dry-cured ham, titin is an important protein responsible for muscle structure and is involved in the connection between actin and myosin. The degradation of this protein was assessed over the curing process, and titin-derived peptides  have been proposed as candidate markers for processing time . MoreoveThe hydrolysis of food proteins and modifications are produced as a result of the cooking process. In the case of pork, a high cooking temperature leads to a reduction in digestibility with significantly different peptidomes. It seems that sites of trypsin cleavages remain more stable with temperature than pepsin cleavages . This de2− was measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method by the hydrolyzation of myosin, troponin, and actin [On the other hand, the search for bioactive peptides from vegetable and animal products is also carried out in cured pork products . Bioactind actin . (LC-MS/MS), or Isobaric tags for relative and absolute quantitation (iTRAQ) should be employed to enhance the efficiency of our quest for protein biomarkers and further validate previous results. Other technical improvements in pork processing were assessed from a proteomic perspective, providing an insight into protein modifications. Peptidomic profiles could further offer an overall overview of the protein digestibility and bioavailability that determine the effect of protein fraction on human health. In the framework of data analysis, we expect that statistics will play a great role in future proteomics, especially in handling the huge data produced by different proteomics methods. Regarding this, we expect that there is great interest in combining different omics techniques in the framework of multi-omics to study the interplay between different macromolecules in relation to the pork phenome. Indeed, phenomics or high-throughput phenotyping is becoming a reality in livestock production systems including pork, and we expect that this global approach will play an important role in the next years and decades. Proteomics is an emerging technology for the rapid and sensitive identification of biomarkers aiming to assess the potential quality of pork products and the impact of food processing technologies. Genetic and rearing conditions influencing technological and sensory meat quality provoke different biochemical and molecular reactions that are regulated by several proteins and pathways including metabolic enzymes. Furthermore, pork quality determined by tenderness, color, drip loss, and intramuscular fat is conducted by structural and sarcoplasmic proteins. In this regard, proteins play a key role in the textural and sensory quality of pork fresh, showing the importance of the study of muscle proteome in pork. The most relevant quality traits were assessed by gel and mass spectrometry analysis. Gel-based proteomics are widely used for the search of protein biomarkers of these quality traits. Even the most sensitive gel-based methods such as protein labeling with fluorescent dyes as fluorescence difference gel electrophoresis (DIGE) were considered. However, gel-free alternatives such as Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH-MS), Liquid chromatography–mass spectrometry/mass spectrometry


Breastfeeding is indicated to support neonatal immune development and to protect against neonatal infections and allergies. Human milk composition is widely studied in relation to these unique abilities, which has led to the identification of various immunomodulating components in human milk, including various bioactive proteins. In addition to proteins, human milk contains free amino acids (FAAs), which have not been well-studied. Of those, the FAAs glutamate and glutamine are by far the most abundant. Levels of these FAAs in human milk sharply increase during the first months of lactation, in contrast to most other FAAs. These unique dynamics are globally consistent, suggesting that their levels in human milk are tightly regulated throughout lactation and, consequently, that they might have specific roles in the developing neonate. Interestingly, free glutamine and glutamate are reported to exhibit immunomodulating capacities, indicating that these FAAs could contribute to neonatal immune development and to the unique protective effects of breastfeeding. This review describes the current understanding of the FAA composition in human milk. Moreover, it provides an overview of the effects of free glutamine and glutamate on immune parameters relevant for allergic sensitization and infections in early life. The data reviewed provide rationale to study the role of free glutamine and glutamate in human milk in the protection against neonatal allergies and infections. Human milk is widely recognized as the best source of infant nutrition. It provides the infant with a highly diverse mix of nutrients that supports optimal development. The health benefits of human milk, however, go beyond that of providing nutrients. An increasing body of evidence suggests that human milk provides the neonate with a protection against a variety of immune-related conditions. For example, it is shown consistently that infants who were exclusively breastfed were less likely to develop respiratory and gastrointestinal infections than infants who fully or partially received an infant milk formula –5. This H1) immune responses and skewed T cell responses to antigens toward T-helper 2 (TH2) immunity. These characteristics correlate with an increased susceptibility to infections and allergies in the neonatal period  in human milk, 5–10% is present in free form. The FAAs glutamate and glutamine are by far the most abundant, both in absolute sense and relative to their protein-bound form, together comprising almost 70% of all FAAs present in human milk . Their lIt is well known that protein quality and quantity are key aspects of the nutritional value of human milk. The total amino acid (TAA) composition of human milk, including protein-bound AAs and FAAs, is used to evaluate the quantity and the quality of the milk proteins and hence is well characterized , 37. HowFAAs in human milk have been reported to account for ~5–10% of the TAA content , 36. DesWhile human milk directly supplies infants with FAAs, human milk proteins could also provide the infant with FAAs via proteolysis in the neonatal gastrointestinal tract. However, the contribution of proteolysis of human milk proteins to the FAA supply of infants might be relatively low, as (complete) proteolysis of these proteins in infants is shown to occur to a minimal extent –52. FactThe unique abilities of FAAs compared to protein-bound AAs and the relatively inefficient proteolytic capacity of neonates underline the importance of understanding the FAA composition in human milk, separate from the TAA composition.The composition of human milk is known to be dynamic over the course of lactation. The total protein content has been consistently shown to decrease in the first 3 months of lactation , 36. It The underlying mechanisms regulating the dynamics of FAA levels in human milk are poorly understood. Cells of the mammary gland secrete proteases and anti-proteases into human milk that together regulate the cleavage of specific AAs from human milk proteins, generating FAAs and peptides . Thus, iTo better understand the mechanisms underlying the secretion of FAAs in human milk, several studies examined the influence of maternal characteristics on the FAA composition in human milk. Whereas, FAA levels seem to be independent of the mothers' age , maternaAll together, these findings indicate that selective FAA transport occurs in mammary tissues during lactation and that levels of FAAs in human milk might be highly regulated throughout lactation.The FAAs glutamine, glutamate, glycine, serine and alanine in human milk have consistently been shown to increase in the first 3 months of lactation, whereas the levels of most other FAAs remain relatively stable along lactation , 55, 56.Studies investigating associations of FAAs with infant anthropometrics are scarce but do report consistent findings. It was recently reported that free glutamate levels in human milk were significantly higher for term infants that had faster weight gain . MoreoveThe finding that levels of free glutamine and glutamate in human milk are relatively high, display unique dynamics along lactation, and are associated with infant anthropometrics urges the need to understand the functions that these FAAs could have during infant development.In the last decade, it has been recognized that glutamine and glutamate are essential AAs at key times in life, including the neonatal period when rapid growth occurs , 79. AltIt is well-established that glutamine and glutamate are important energy substrates for intestinal epithelial cells (IECs) and immune cells, especially during periods of rapid growth . In factBesides serving as energy substrates, free glutamine and glutamate are both specific precursors for glutathione, which is the main antioxidant in IECs and immune cells and critical for the prevention of cellular damage caused by pro-oxidants . An imbaIn the rapidly growing neonate where the intestines are not yet fully developed, it is crucial to achieve and maintain rapid growth of IECs. Moreover, it is well-established that intestinal barrier function is a crucial factor in the protection against allergies and infections, by preventing allergen and bacterial translocation from the gut lumen into the immune cell-populated lamina propria and mesenteric lymph nodes –107. In in vitro. For instance, glutamine dose-dependently enhanced cell proliferation and differentiation of neonatal porcine and rat IECs, through activating multiple mitogen-activated protein kinases (MAPKs)  –110. Mor (MAPKs) , 111–113 (MAPKs) –116. Rem (MAPKs) .in vitro studies with neonatal porcine and human adult IEC lines have revealed that glutamine restriction reduces the expression of the major tight junction proteins, including claudin and occludin proteins, which are vital for intestinal barrier function  immune activity (H1) immunity is highly limited and gradually develops during the postnatal period , intestinal mast cells, peripheral mononuclear cells (PBMCs) and monocytes , or a decreased colonization of Proteobacteria and Bifidobacteria are shown to be at increased risk of developing food allergies (H2 and TH1 immunity (Accumulating evidence indicates that the gut microbiota plays a vital role in tolerance induction to dietary antigens –183. Accllergies –191. Mecimmunity , 193. Thimmunity . Thus, hFirmicutes in the jejunum and ileum, and decreased the Firmicutes-to-Bacteroidetes ratio in the ileum (Clostridium spp. in the ileum, of which colonization in early life has been associated with increased risk of allergic diseases (Bifidobacteria in the jejunum of healthy weaned mice (The ability of dietary glutamine to modify the microbiota composition is shown in various young animals. A study in weaning mice demonstrated that dietary glutamine decreased the content of he ileum . Similarhe ileum , 196. Indiseases , 198. Fined mice , and to ned mice , 199.Firmicutes-to-Bacteroidetes ratio in the ileum, although this effect was only seen when given in combination with a high fat diet and was not observed in other intestinal sections. Perhaps more interestingly, dietary glutamate specifically promoted the colonization of prausnitzii and Faecalibacterium prausnitzii in post-weaning pigs (Roseburia in early life has been positively associated with the acquisition of tolerance to cow's milk (Faecalibacterium prausnitzii is indicated to play a role in the prevention of food allergy (To our knowledge, only two animal studies examined the effects of dietary glutamate on the intestinal microbiota composition to date, both of which used animals in their post-weaning phase. It has been reported that dietary glutamate markedly enhanced the bacterial diversity in the intestinal flora of healthy post-weaning pigs . Moreoveing pigs , 201. Thw's milk , and Fae allergy –205. The allergy . Butyrat allergy –209.Research indicates that breastfeeding during the first months of life provides protection against immune-related conditions in neonates and later in life. These conditions include gastrointestinal infections and several allergic diseases including food allergy. It is indicated that the transfer of specific immunomodulating components, such as bioactive proteins, from mother to infant through human milk contributes to this protective effect. In addition to proteins, human milk contains FAAs, which have unique characteristics. They are more readily absorbed than protein-derived AAs and can be recognized by specific receptors on various cells. Moreover, whereas protein-bound AAs decrease during the lactation period in a non-AA-specific manner, temporal changes of FAAs in human milk are highly AA-specific. These dynamics in FAA levels are globally consistent and thus seemingly independent of ethnicity, demographics and maternal diet. This suggests that selective FAA transport occurs in the mammary gland, that FAA levels in human milk are strictly regulated and, consequently, that FAAs are likely to be of physiological relevance in the developing infant.H1 promoting and/or TH2 inhibiting effects on various intestinal immune cells, and modify the abundance of gut microbiota that might play a role in allergic sensitization (With regards to individual FAAs in human milk, free glutamine and glutamate display particularly remarkable characteristics. They account for almost 70% of the FAA content in human milk, they both drastically increase in the first 3 months of lactation and their levels have been shown to positively correlate with infant growth, suggestive of important functions in the developing neonate. In neonates, dietary glutamine and glutamate are mainly used by the intestines. Remarkably, studies in neonatal immune cells and young animals demonstrate that these FAAs can have a wide range of effects on cells in developing intestines, also at concentrations similar to their levels in human milk. In short, they are reported to increase the growth of intestinal epithelial cells, enhance intestinal barrier function, influence immune cell development and populations in the gut-associated lymphoid tissue, exert anti-inflammatory and potentially Ttization . TogetheAll together, the findings described in this review warrant further research into the contribution of free glutamine and glutamate in human milk to the protection against neonatal allergies and infections. Levels of free glutamine and glutamate, in addition to that of other bioactive factors that could influence early life immune development, are considerably higher in human milk than in standard infant milk formulas, leading to significant differences in the intake of these FAAs between breastfed and formula-fed children –212. As JS, AH, and SW: conceptualization and literature searches. AH, SW, and JG: supervision. JS: writing original draft. JS, AH, SW, and JG: review and editing. All authors read and approved the manuscript.SW is current employee of Danone Nutricia Research. JG is part-time employee of Danone Nutricia Research and Utrecht University. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


This review focuses on synthetic and natural amphiphilic systems prepared from straight-chain and macrocyclic compounds capable of self-assembly with the formation of nanoscale aggregates of different morphology and their application as drug carriers. Since numerous biological species  bear negatively charged fragments, much attention is paid to cationic carriers providing high affinity for encapsulated drugs to targeted cells. First part of the review is devoted to self-assembling and functional properties of surfactant systems, with special attention focusing on cationic amphiphiles, including those bearing natural or cleavable fragments. Further, lipid formulations, especially liposomes, are discussed in terms of their fabrication and application for intracellular drug delivery. This section highlights several features of these carriers, including noncovalent modification of lipid formulations by cationic surfactants, pH-responsive properties, endosomal escape, etc. Third part of the review deals with nanocarriers based on macrocyclic compounds, with such important characteristics as mucoadhesive properties emphasized. In this section, different combinations of cyclodextrin platform conjugated with polymers is considered as drug delivery systems with synergetic effect that improves solubility, targeting and biocompatibility of formulations. Self-assembling systems based on amphiphilic compounds find wide application in different fundamental and practical areas due to their unique ability to form nanoscale assemblies with gradients of polarity, viscosity, electric charge and other characteristics ,2,3. In Among amphiphilic compounds, special attention is received by cationic surfactants, which is related to both their fundamental and practical relevance. Researchers have succeeded in designing and synthesizing a variety of homologous series of cationic surfactants differing in the structure of head groups, which allows for determining the role of the polar fragment in the physicochemical properties and functional activity of supramolecular systems based on cationic surfactants in terms of structure–properties–activity relationships. Numerous fundamental approaches and practical applications involving cationic surfactants have recently been demonstrated in the fields of biochemistry, nanomedicine, pharmacy, catalysis, corrosion protection, oil recovery, food, cosmetics, etc. . These bRecently, much attention has been received by the so-called cleavable surfactants bearing ester, amide, disulfide or other degradable fragments . While rA series of publications ,16,17,18® were developed with the use of a series of cationic surfactants. Carbamate-bearing surfactants were found to show the highest activity, with a 3-fold increase in the Lontrel® concentration observed in the plants tested and improved wetting ability compared to an unformulated preparation. These results could be further enhanced by increasing the temperature. The biomedical functionality of carbamate-bearing surfactants could be significantly modified by the introduction of an imidazolium fragment MicrococcStaphylococcus aureus [Below the Krafft point, the amphiphiles self-organized into tubular structures of various morphologies forming hydrogels at low surfactant concentration; above the Krafft point, there was a transition from tubular structures to micelles or vesicles, depending on the structure of the surfactant, and this transition was thermo-reversible in the physiological temperature range. The synthesis of histidine-based surfactants with high antimicrobial activity against Gram-positive and Gram-negative bacteria is given in , with sos aureus . The auts aureus  investigs aureus . The CMCSeveral examples of amino acid-based amphiphiles and their properties are given in Another promising approach to design substances meeting biomedical application criteria is the functionalization of amphiphiles by a saccharide moiety , that maThe majority of monosaccharide amphiphilic compounds are represented by glucose derivatives. Interestingly, the selection of functionalization of glucose-containing amphiphilic compounds and using of various approaches to molecule design could become a useful tool for tailoring desired practical properties. For example: (i) significant foamability and high emulsion stability for toluene/water system could be achieved in the case of application of amido- and/or alkoxy derivatives of this hexose ; (ii) inDisaccharide derivatives cover amphiphiles mainly containing lactose, maltose and sucrose fragments in their structures. The presence of a lactose moiety in the molecule provides both industrial and biotechnological significance to these compositions. These compounds could be used as efficient micellar catalysts of Ullmann C-S coupling reactions in water  ; highly-There are only a couple of recent applications dealing with trisaccharide amphiphilic compounds, nevertheless compositions based on them show themselves to be biocompatible and suitable for biotechnological purposes. For example, raffinose and melezitose amphiphilic derivatives was documented as effective solubilizers for polycyclic aromatic hydrocarbons possessing acceptable hemolytic activity, which could be reduced by shortening the amphiphile alkyl tail  as well A series of alkyltriazole glycosides (ATGs) with different length of spacer linking the sugar and triazole moieties were synthesized, and their surface-active properties and phase behavior were studied . It was In  the syntThe effect of three different mannose-based surfactants included in the lipid bilayer on the physicochemical parameters of the system was studied in . It was In  the effeGemini surfactants containing two hydrophilic head groups and two hydrophobic tails exhibit beneficial physicochemical properties, in particular, they have significantly reduced CMC values and increased surface activity compared to monomeric amphiphiles. In the works of Liu et al. , gemini In  glucono-Cationic surfactants are mentioned above to be an important class of chemical compounds, which find wide practical application as emulsifiers in cosmetology, bactericidal agents, corrosion inhibitors, antifoams and in medicine. At present, many studies are focused on the increase of biocompatibility and biodegradability of cationic surfactants. To achieve these goals, sugar-based cationic surfactants are obtained. Glucose-based cationic surfactants containing an ester group were obtained in , and theIn  reverse Analysis of existing representatives of sugar-based amphiphiles has demonstrated that key criteria responsible for the best aggregation characteristics  of sugar-based surfactants are the presence of more than one hydrophobic tail in the structure and the transition from monosaccharide to oligosaccharide derivatives. If the first mentioned criterion is very similar to conventional surfactants and has the same nature, the second one could be due to allowing for more intermolecular hydrogen bonds between the OH-groups of additional saccharide moieties. Several examples of amphiphiles with superior self-assembling propensity are listed in Salts of bile acids are derived from steroid acids contained in the bile of mammals. Their main difference from conventional amphiphiles is the presence of a bulky scaffold, which predetermines their unique properties. For example, bile salts are known as effective transporters of lipids , regulatp-tertbutylbenzoyl derivatives of cholic acid, which when mixed with a copolymer and in a covalent conjugation with tripeptide demonstrate fabrication of thermoresponsive supramolecular structures [Further attempts of bile acids functionalization have been presented in recent reports. There are a couple of reports discussing ructures  and twisructures  as well ructures .A special place among amphiphiles bearing natural fragments is occupied by the so-called nucleolipids–hydrophobic derivatives of nucleobases, nucleosides and nucleotides. Interest in these compounds is mostly due to the nitrogenous bases  inserted in their chemical structure and capable of being involved in additional specific interactions. As an example, nucleolipids was known by their capability to form films at air/liquid interfaces , coatingA comparative analysis of collected data for nucleolipids of various structure  was condSilicone-based surfactants are new effective amphiphiles with advanced characteristics. They are widely used in the leather industry , agriculThe presence of a tetrasiloxane fragment in the structure of cationic gemini surfactants can potentially improve the biocompatibility and various characteristics of amphiphiles. A series of new cationic gemini surfactants with a tetrasiloxane fragment a was synTwo tetrasiloxane-based imidazolium gemini surfactants with methylene spacer groups b were sy2SO4) on aggregation of these surfactants was studied.A series of organosilicon bis-quaternary amphiphiles was prepared in . It was nH2n+1, n = 8,10,12,14,16,18) was obtained in [The authors of  obtainedained in . It was In  a seriesA four-step synthesis and characterization of a new silicone-containing polyurethane surfactant are given in . The resThe above literature survey demonstrates that the synthesis of novel amphiphilic compounds and investigation of their properties and practical applications have attracted much attention from researchers. Many efforts have been directed on the improvement of the surface activity and adsorption characteristics of surfactants and solubilization capacity of their assemblies, since these are basic properties responsible for the bio- and nanotechnological potential of supramolecular systems. In the case of surfactants with natural or cleavable fragments special attention has been paid to their applications in pharmacy and medicine due to their reduced toxicity, biocompatibility and diverse morphology, with most interest being attracted by cationic surfactants. These amphiphilic compounds are further considered as building blocks and modifiers for nanocarriers capable of enhancing their efficacy as drug delivery systems.As illustrated above, amphiphilic systems are of particular interest in biomedical applications , which cImportantly, the physicochemical properties, e.g., size, shape, charge characteristics of nanocarriers are key factors determining their biological fate, especially the primary interaction with cell membranes ,174. In Usually nanoparticles cannot passively pass through the hydrophobic cell membrane. Thus, the cell membrane presents the first barrier to their penetration into the targeted intracellular compartment. At the same time, it is well known that several active intracellular uptake pathways that could be used for nanoparticle internalization are available at the cell membrane. During the first step of intracellular uptake, the cell membrane forms an invagination to take up a portion of extracellular fluid including cargo molecules in it. This active intracellular transport, which is called endocytosis, for cargos smaller than 200 nm can be basically classified into three major categories: receptor-mediated clathrin-dependent endocytosis, caveolar pathway, and the less understood panoply of clathrin- and caveolae-independent pathways .Receptor-mediated clathrin-dependent endocytosis is the mechanism used to internalize cargo into the cells via binding to their specific receptors on the surface of the cell membrane. After formation of these ligand-receptor complexes, the cytosolic protein named clathrin-1 polymerizes on the inner surface of cell membrane and the cargo is engulfed through the formation of clathrin-coated vesicles.Caveolae-mediated endocytosis is the mechanism of active intracellular transport which involves membrane invaginations called caveolae. Caveolae are formed by assembly of membrane proteins caveolins. It is important to note that endocytosis via caveolae remains poorly characterized, and it is not clear whether caveolae-mediated endocytosis is constitutive, as clathrin-dependent pathway.There are also a number of pathways for endocytosis, where specific proteins that coat vesicles, such as clathrin or caveolins, were not identified. It is known that endocytosis occurs in cells, when clathrin-dependent endocytosis was inhibited by hypotonic shock, cytosolic potassium depletion, or acidification of cytosol . HoweverThis information about the main pathways for endocytosis of nanoparticles has been successfully used to develop new nanocarriers for targeted delivery of drugs. For example, mechanisms of receptor-mediated endocytosis can be used for targeted delivery of anticancer drugs. It has been shown that density of certain receptors that are internalized via clathrin-dependent endocytosis is much higher in cancer cells than in normal cells. Functionalization of nanoparticles by the ligands to these receptors can promote both specific binding to the particular cancer cells as well as the intracellular uptake of nanoparticles. The commonly used ligands for anticancer targeted drug delivery are transferrin receptor ligands , folate Following internalization, the next crucial step for targeted drug delivery is the intracellular trafficking for localization in the target cellular compartment. It is important to note that in most cases, initially, the intracellular localization of the nanoparticles is not optimal. After internalization of nanoparticles via clathrin-dependent endocytosis or clathrin- and caveolin-independent endocytosis, they will first fuse with early endosomes, which will transform into late endosomes upon the maturation. The late endosomes will then fuse with lysosomes and then nanoparticles will be digested by the hydrolytic enzymes activated at acidic pH occurring in these microdomains. Thus, to reach the targeted intracellular compartment, nanoparticles must be able to escape this endosome-lysosome pathway, bypassing the degradation process. Over the past two decades, to prevent the endosomal degradation different strategies have been developed. It was shown that endosomal escape of nanoparticles into the cytoplasm can be promoted by the incorporation of cell membrane penetrating peptides  or pH-deFor lipid formulations such as liposomes, modification with cationic or fusogenic lipids has been applied. Cationic lipids destabilize negatively charged endosomal membranes through ion-pairing mechanism, which involves changes in geometry of amphiphilic matrix from cylinder to reverse cone, thereby inducing the formation of reverse hexagonal phase . SimilarMechanisms of endosomal escape mentioned above are postulated for other organic nanoparticles, including those based on polymeric building blocks. For polymers  bearing ionogenic groups capable of being protonated at acidic pH, additional mechanism is assumed referred to as proton-sponge effect. Generation of positively charged centers in macromolecules is accompanied by accumulation of counterions within the endosomal compartment, thereby inducing rupture of biomembranes due to increase in osmotic pressure and swelling effect ,158. MeaDevelopment of nanocarriers that use caveolae-mediated endocytosis for their internalization also can be considered as a perspective approach to escape from endosome-lysosome pathway. It was shown that caveolae can fuse with early endosome but also with “caveosome”. Caveosomes are the cell compartments that exist at neutral pH and do not have hydrolytic enzymes . Thus, nIt should be noted that not only the endo-lysosomal system affects the nanoparticle but also vice versa. In recent studies, it was shown that cells treated with nanosized fluvastatin show increasing levels of autophagosome formation that competitively inhibit lysosomal activity . It was Among the clinically relevant nanocarriers, liposomes have a special place, since only liposomal formulations of antitumor drugs were first approved for clinical practice. This can be exemplified by liposomal DOX preparation, Doxil, which was the first liposomal drug approved by FDA, which was followed by the liposomal vincristine carrier Marqibo and liposomal daunorubicin drug DaunoXome, etc. ,189,190.In the history of liposomal drug delivery systems, a lot of ways of improving therapeutic efficacy of drugs have been found ,191, as In turn, pegylated liposomes along with the stabilization effect on nanoparticles resulted in a body of concerns referred to as the PEG dilemma, with limitations on cellular uptake and further endosomal escape, as well as accelerated blood clearance effect (ABC) occurred. The pegylation significantly lowered the liposomes’ interaction with the target tissue alongside opsonins. To overcome this problem, the next generation of liposomal formulations were developed ,194. AmoConcerning the different aspects of incremental advances in design and practical application of liposomal nanocarriers, a variety of liposomal drugs are on the market, and much more are in different stages of clinical trials, which is comprehensively discussed in reviews ,189,190.A survey of recent literature allows us to provide a brief outline of the urgency of the studies focusing on CLs. One of the most studied and recognized sphere of CLs application is their use for transfection. Their positive charge makes them an exceptional nanocarrier for negatively charged nucleic acid sequences and is advantageous due to higher tissue uptake and endosomal escape capabilities . For a fOne of the well-known downsides of the CLs is their toxicity, which is inherent to the positive charge; thoughtful dosage can mitigate the toxic effects, as shown in the recent work by Knudsen and co-workers . AnotherMeanwhile, attempts are being made to investigate CLs as delivery vehicles. The cationic surface charge is very beneficial when dealing with endosomal trafficking due to the negative charge of endosome membrane ,209. TreTo summarize this brief discussion, cationic liposomes are of current importance in view of their high biomedical potentiality. Nevertheless, in some cases large positive charge of nanocarriers may prevent their efficacy due to, e.g., non-specific membrane fusion with other than endosomal membranes, enhanced opsonization or agglutination effects, elevated toxicity of formulations, etc. Therefore, strategies of fabrication of the partially charged liposomes and/or charge-switchable nanocarriers are of importance. As an alternative to the formation of mixed compositions based on phosphatidylcholines and cationic lipids, a strategy of non-covalent modification of the surface of liposomes by cationic surfactants is actively developing . Among tIn the formation of modified liposomes, the degree of oligomerization of a cationic surfactant  and the ratio of lipid to surfactant are crucial factors, since only at a low surfactant concentration the incorporation of surfactant into the lipid bilayer of liposomes is possible, while after a certain threshold value complete destruction of the lipid bilayer and the formation of mixed surfactant-lipid micelles are observed ,218,219.H)-one, when encapsulated in imidazolium surfactant-modified liposomes shows a sharp decrease in viability of diseased cells (HeLa) and increase of viability of normal cells (Chang liver) [Escherichia coli with high inhibitory activity. The investigation on liposomes formulated with 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and cationic surfactants: -N-hexadecyl-N-methylprolinolinium bromide, -N-hexadecyl-N-methylprolinolinium bromide, N-hexadecyl-L-prolinolinium hydrobromide, N,N-dihexadecyl-L-prolinolinium bromide, in the presence and in the absence of m-tetra-hydroxyphenylchlorin is presented in [Staphylococcus aureus bacteria and one of them has shown results comparable to free chlorin. The traditional cationic surfactant CTAB was used in the work to impart a positive charge to niosomes based on N-myristoylserinol and cholesterol [We begin our consideration of this issue with monomeric single-tailed surfactants . Pegylatg liver) . Cationig liver) . Hybrid g liver) . It was g liver) . Mixed lg liver) . Modifieented in . Differelesterol . In the Staphylococcus aureus and Streptococcus pyogenes growth in vitro. Most persistent penetration of human skin ex vivo was achieved with use of Curcumin-CLs.Very often, long-chain amines are used to modify the surface of liposomes, especially when developing compositions for treating skin diseases. Hexadecylamine was integrated into the liposomes based on DPPC and cholesterol (8:2 (mol)) obtained by microfluidic assembly under conditions of optimization of flow rate to increase colloidal stability . AdditioBetween monomeric and gemini surfactants, an intermediate position is occupied by surfactants with two hydrophobic tails and one head group, which can form vesicles instead of micelles in a solution and have an ability to integrate into a lipid bilayer . For exaThe next interesting block about modified liposomes is associated with gemini surfactants . ModificStaphylococcus aureus (MIC = 7.8 μg·mL−1) and lowest cytotoxicity (IC50 > 125). The resulting composition was tested for transdermal drug delivery. Also, for transdermal delivery, niosomes were obtained using lysine-based gemini surfactant and cholesterol [The liposomal formulation based on phosphatidylcholine and the tetradecyl gemini derivative DABCO showed the highest antibacterial activity against lesterol . The mosS,R)-2,3-dimethoxy-1,4-bisbutane bromide and on their ability to deliver the plant alkaloid voacamine to U-2 OS/DX osteosarcoma cells was carried out. The effect of incorporating surfactants into the lipid bilayer on the efficiency of encapsulation depends on the nature of the lipid itself. A series of papers by this team of authors ends up in preparation of pegylated gemini-modified liposomes [Interactions of surfactant modified liposomes with plasma membranes are elucidated in ,236, wheiposomes .In  the authThus, according to the increasing number of articles devoted to the production of cationic liposomes by non-covalent surface modification with surfactants, we can say that the technique is successful and relevant. By selecting the structure of the lipid and surfactant, the ratio of the components, it is possible to control the efficiency of encapsulation and loading, the release rate of the substrate, the ability of the liposomal formulation to overcome biological barriers  to obtain commercially available nanocontainers. pH-responsive liposome formulations present a very vast range of benefits and have been studied since 1980s and especially in the last decades as candidates for second-generation drug delivery nanoparticles, but they are yet to be approved for clinical applications . One wayAnother unique feature of such nanocontainers can help solve the problem of premature clearance from bloodstream , pH-respIn the above discussion, we focus on the beneficial applications of nanocarriers bearing cationic charges, focusing on the family of lipid formulations modified with surfactants. At the same time, in the field of organic nanoparticles as drug delivery systems (nanomedicines) the special class of biomimetic transporters of macrocyclic nature and polymeric nanocarriers, especially those of cationic nature, play a significant role which remained unaddressed in the review. Therefore, the final part focuses on supramolecular assemblies based on mixed macrocyclic and polymeric systems, with special attention paid to cationic polymers due to their enhanced mucoadhesive properties. Among the various methods of delivery and modification of dosage forms, the phenomenon of mucoadhesion, i.e., the ability of the compositions to adhere to the surface of the mucous membrane (mucose) is of particular importance for increasing the drug bioavailability. In order to keep the encapsulated drug in the tissues of the body  for a long period of time, the drug nanocontainer should have mucoadhesive properties. The lack of adherence to these tissues is the main problem observed in clinical trials with drugs. Hence, the understanding of the mechanisms of mucoadhesion with mucus is of paramount importance for creating effective systems for the delivery of drugs to the human body. In this line of research, special attention is paid to the modification of systems by water-soluble and water-swelling polymers, which can improve mucoadhesive properties in order to increase the therapeutic effect . In thisIt is well known that cyclodextrins (CDs) are able to increase the solubility of various lipophilic molecules by forming relatively stable inclusion complexes. The stability of these complexes is usually higher, when the size of the guest molecule is suitable for the size of the macrocycle cavity. Due to the receptor action and low toxicity, CDs are widely used in the design of drug delivery systems. Chemical modification of CDs allows the formation of derivatives with improved physicochemical properties  as compared to natural CDs, which can positively affect their biomedical efficiency . This moThe formation of inclusion complexes of CDs accompanied by an increase in the aqueous solubility of drugs can lead to spontaneous self-assembly of these complexes. Encapsulation of the drug telmisartan in the γ-CD cavity can initiate the aggregation of inclusion complexes. The γ-CD complexes with telmisartan are able to aggregate into spherical nanoparticles with diameters from 50 nm to 500 nm . HoweverHydroxypropyl methylcellulose and other water-soluble polymers  can be used not only for stabilization, but also to increase the solubilization of the drug as an additional agent to CDs. The ternary CD–polymer–celecoxib complexes form joint aggregates ranging from 250 to 350 nm in size and display a large solubilization capacity for the drug . Eye droIn  the abilObviously, the introduction of CD fragments into the polymer chain of chitosan can lead to the formation of a molecular carrier with encapsulating properties for drugs, transport properties through biological barriers and the ability to control drug release. However, it is not always possible to solve the problem of the low solubility of chitosan in water by crosslinking of CD with the polymer; therefore, the chitosan was quaternized ,250. ConPolymeric surfactants may also be involved in the development of mucoadhesive systems. Pluronics were used to improve the delivery of itraconazole to the eyes . The surThe combination of hydroxypropyl-β-CD with Tween 80 makes it possible to stabilize the drug econazole nitrate, protecting the encapsulated drug from aggregation for one year at room temperature . This coHydroxypropyl-β-CD and polyvinylpyrrolidone were also involved in the creation of rapidly dissolving nanofibers capable of encapsulating clotrimazole . These nCDs can be used not only to increase the water solubility of drugs, but also to improve their taste. Hydroxypropyl-β-CD eliminates the unpleasant taste of irsogladine maleate used for treatment of stomatitis. Addition of the macrocycle to the mucoadhesive gum ghatti, consisting of high-molecular-weight polysaccharides, leads to the formation of a viscous solution as a result of aggregation due to hydrogen bonds between the macrocycle and the polymer .Mucoadhesive microspheres based on a complex of γ-CD and raloxifene can be obtained using various proportions of carbopol and hydroxypropyl methylcellulose. The size of the obtained particles on the percentage of polymers ranged from 3 to 15 microns, and the encapsulation efficiency of raloxifene ranged from 81.63 to 87.73%. These microspheres had remarkable mucoadhesion and controlled drug release lasting up to 24 h. More than 60% of the oral dose was kept for 4 h in the stomach of Wistar rats .The mixed compositions based on CDs and polymers can be used not only to encapsulate drugs, but also proteins and enzymes. In  microsphNanoparticles based on hydroxypropyl-β-CD and chitosan can be used in targeting various organs of the gastrointestinal tract. A study of the release of hydrophobic substrate from this composition showed a sharp release (more than 40%) in the gastric fluid due to the breakdown of chitosan and β-CD inclusion complex at low pH . As in tDillenia indica [Dillenia polymer. In addition, Dillenia has sufficient mucoadhesive properties and good compatibility with the drug or any other ingredients of the composition.Besides a wide range of different nanoparticles and microspheres, mucoadhesive micro- and nanocapsules can be obtained from macrocycles and polymers. Simvastatin microcapsules were obtained by complexation with hydroxypropyl-β-CD, hydrophilic sodium alginate and mucilage obtained from a indica . Drug reAmphiphilic CDs and polymers can be used to create mucoadhesive drug nanocontainers. β-CD with hexyl tails forms nanocapsules with poly-ε-caprolactone for encapsulation of camptothecin . Drug loThe addition of hydroxypropyl-β-CD as a permeation enhancer improves the penetration of timolol maleate through the bovine cornea . HoweverNon-covalent aggregation of macrocycles with polymers was used to develop mucoadhesive compositions with effective encapsulating ability. Ion cross-linked nanoparticles based on oppositely charged chitosan and sulfobutyl ether-β-CD are an interesting system for the delivery of econazole nitrate to the mucous membrane of the eye . Half ofThe use of complexation between α-CD molecules and polysaccharides modified with hydrophobic alkyl chains leads to the spontaneous formation of nano- or micron-sized platelets that can encapsulate antimicrobials for effective treatment of mucosal infections ,267. TheIn addition to crosslinking of macrocycles on flexible open-chain molecules, β-CD can be used to modify the mesoporous silica nanoparticles with hydroxyl, amino, and thiol groups . Thiol-fThe authors of  definitem-hydroxyphenyl)porphyrin can be modified with polyethylene glycol or chitosan [For modification of mucoadhesive nanoparticles not only CDs can be used, but also derivatives of porphyrins, which are a promising choice for drug delivery systems in photodynamic therapy, combining the advantages of efficient drug transport to target cells and reducing of undesirable side effects. The surface of nanoparticles based on biodegradable poly-DL-lactide-co-glycolide and 5,10,15,20-tetrakis and environmentally friendly silicone moiety approves oneself as a promising strategy for construction of soft supramolecular systems with desired shape and stimuli-responsive functions. Specifically, this strategy has a great possibility for development of drug delivery systems with various morphology, tunable physicochemical properties and stability, which could become a useful tool for the solution of the problem of controlled release of cargo.Liposomal formulations have received the main attention due to the fact that they were among the few carriers approved for the clinical treatment. Since numerous marketed liposomal therapeutics are available, these formulations appeared to be of particular importance for the analysis of bench-to-bedside factors. Several generations of liposomes were designed and actively advanced towards the market, beginning with conventional liposomes to stealth and targeted modifications, with the strategies for enhanced loading capacity, triggered release, endosomal escape, solution of PEG dilemma, and overcoming the BBB developed. Although specificity occurs in the development of different family of carriers, there are many common challenges typical for each of them. This is exemplified herein by such problem as endosomal escape that was discussed in detail, including ways for overcoming it. As alternative nanocarriers solid lipid nanoparticles, niosomes and cerasomes are identified that are assumed to provide the better standard of toxicity, biocompatibility, loading capacity and morphological stability in the near future. Analysis of the reasons preventing fast progress in the translation of the formulated drugs to the clinic reveals that they are mostly related to the manufacturing process and consist of the loss of product stability and quality upon scaling up of the preparation procedures, as well as in problems with the reliability and reproducibility of the final drug. To some extent, these drawbacks can be reduced by simplification of the preparation of the nanocarriers, excluding complicated synthetic procedures. Therefore, much attention is paid to the publications focusing on the noncovalent modification of nanocarriers, with the priority given to cationic surfactants, that revealed their beneficial efficacy from the viewpoint of colloidal stability, overcoming the biological barriers, cellular uptake, mitochondria-targeted drug delivery, etc. In this context, modification of nanocarriers with cationic additives is a promising strategy, especially with the use of charge-reversal amphiphiles that allow for preventing of lysosome degradation of encapsulated cargo. The disadvantage of cationic carriers is their toxicity that can be solved by tuning the lipid composition, with the balance between toxic characteristics and beneficial properties  achieved. As a whole, the key point that should be stressed is the tendency to engineering the polycomponent formulations providing a synergetic effect. Hence, the colloidal, polymer and supramolecular approaches must be jointly implemented in order to find new effective nanocarriers for the successful delivery of molecules needed by the body. From this viewpoint, macrocycle–polymer compositions are of importance, since macrocycles exhibit selective binding of the drug resulting in enhanced solubility, while polymer shell guarantees the stability in biomicroenvironment and improved affinity to biointerfaces. Since the physicochemical and biological properties of some pure surfactants, mucoadhesive polymers and macrocycles, as well as the published results of their clinical use, present them as favorable raw materials for the preparation of medicinal compositions, the mixed compositions based on them can be considered promising for the design of effective drug nanocarriers.