Circular RNAs have a unique covalent closed-loop structure, which is mainly formed by the reverse splicing of exons from a precursor mRNA. With the development of key technologies such as high-throughput sequencing and the advancement of bioinformatics in recent years, our understanding of circular RNAs has become increasingly more detailed, and their abnormal expression in a variety of cancers has attracted increasing attention. Studies have shown that circSNARCA5 not only plays a crucial role in the occurrence and development of cancer but may also serve as a reliable indicator for tumor screening or a good marker for evaluating cancer prognosis. Nevertheless, there are no reviews focusing on the relationship between circSMARCA5 and cancer. Therefore, we will first explain the main biological characteristics of circSMARCA5, such as biogenesis and biological effects. Then, the focus will be on its role and significance in cancer. Finally, we will summarize the known information on circSMARCA5 in cancer and discuss future research prospects. Circular RNAs (circRNAs) are continuous, single-stranded endogenous RNAs characterized by a closed circular structure without a 5′-terminal cap or a 3′-end poly (A) tail , 2. In 1CircSMARCA5 plays a role mainly through their interaction with other RNAs –9. CircSNext, we will combine the literature to describe the characteristics and potential mechanisms of circSMARCA5 in a variety of cancers, review the results and ideas of multiple studies, provide a basis for further research, and actively explore the significance of circSMARCA5 for cancer treatment and diagnosis.In recent years, numerous studies reported that circRNAs have different degrees of influence on the occurrence and progression of diseases through various mechanisms. In cancer, circRNAs affect cell proliferation, differentiation, and apoptosis, thereby influencing tumor formation and progression. Therefore, the research on the role of circRNAs in cancer is particularly important. SMARCA5  is a member of the ISWI family of chromatin remodelers. It regulates chromosome remodeling through a variety of mechanisms . Like otIn general, pre-mRNAs formed after transcription can be converted into circRNAs. The formation of circRNAs mainly depends on the cutting method, and theoretically, it is controlled by cis-regulatory elements and trans-acting factors. Although the specific mechanism of reverse shearing is not clear, it is certain that reverse shearing requires special tools, and its efficiency is lower than that of standard shearing, resulting in a lower steady-state level of circRNAs compared to their linear counterparts. The efficiency of reverse shearing mainly depends on the shearing sites around the exons. The above-mentioned circSMARCA5 (hsa_circ_0001445) is produced from exons 15 and 16 of the SMARCA5 gene located on chromosome 4q31.21 and has a short length of only 269 nucleotides  10, 16], 1610, 1circSMARCA5 is expressed in a variety of cells . Among tcircSMARCA5 expression exhibits abnormal fluctuations in many cancers. Its expression is reduced in most cancers, such as CRC, MM, ependymoma, NSCLC, intrahepatic cholangiocarcinoma (ICC), HCC, and glioblastoma multiforme (GBM)  11–14, –22, 30. Colorectal cancer is the most common malignant tumor that originates in the colon and rectum, and it remains a leading cause of cancer-related deaths worldwide. Although recent studies have shown that the incidence of colorectal cancer is decreasing in some regions , 34, oveStudies have shown that overexpression of circSMRCA5 can inhibit the proliferation, metastasis, and invasion of colorectal cancer, and these effects can be achieved through different modes of action , 36. FirLiver cancer is one of the cancers with the highest mortality rate in the world . AlthougStudies have pointed out that circSMARCA5 can specifically sponge miR-17-3p and miR-181b-5p. Furthermore, TIMP3 is a common target of miR-173p and miR-181b-5p, which can significantly inhibit the expression of TIMP3. Conversely, circSMARCA5 can increase the expression of TIMP3 by sponging miR-17-3p and miR-181b-5p . TIMP3 hIntrahepatic cholangiocarcinoma is a primary malignant tumor of the liver whose incidence is second only to hepatocellular carcinoma, and has been increasing over the past 20 years , 44, 45.Studies revealed that the expression of circSMARCA5 in cancer tissues was significantly decreased relative to the surrounding tissues, and the level of circSMARCA5 was negatively correlated with the ECOG function score, TNM staging, and CA199 abnormal state, indicating that it can be used as an independent prognostic marker. It was also found that circSMARCA5 can not only inhibit the invasion and proliferation of cancer cells by itself but also increase their chemosensitivity to cisplatin/gemcitabine. This may be one of the factors why circSMARCA5 expression predicts a good prognosis in a variety of cancers .Lung cancer is one of the leading causes of cancer-related deaths worldwide. The two-year relative survival rate of patients with non-small-cell lung cancer has increased from 34% in 2009–2010 to 42% in 2015–2016 . AlthougStudies have shown that circSMARCA5 can not only sponge miR-19b-3p, thereby increasing the expression of HOXA9 , but alsAlthough cervical cancer can be easily screened, more than 4000 women lost their lives due to this malignancy in 2018 alone. Among women aged 20 to 39, it is still the second leading cause of cancer-related deaths . AlthougStudies have shown that SND1 is an RBP that binds to circSMARCA5. SND1 has been proven in other studies to promote the proliferation and invasion of a variety of cancers –50, and Bladder cancer is a malignant tumor that occurs in the bladder mucosa. It is the most common malignant tumor of the urinary system, with 400,000 new cases each year . It was Multiple myeloma is a malignant tumor caused by the abnormal proliferation of plasma cells, often accompanied by multiple osteolytic damage, hypercalcemia, anemia, and kidney damage . AlthougGlioblastoma multiforme is the most common malignant primary brain tumor in adults. It is highly aggressive, and there is still no effective treatment method. The median survival time following diagnosis is approximately 16 months –58. GBM The role of circSMARCA5 in GBM is closely related to serine and arginine-rich splicing factor 1 (SRSF1) , 63, 64.Prostate cancer (PCA) is one of the most common cancers affecting men in the world. The androgen receptor pathway is crucial for the occurrence and development of PCa, and genetic factors are considered to be dominant , 68.Relevant studies indicate that the level of circSMARCA5 is increased in cancer tissues , 32, whiThe important roles of circRNAs in different cancers have received increasing attention. Due to their unique properties and functions, circRNAs are of great significance for the diagnosis, treatment, and prognosis of cancer. For mouse models, antisense of circSMARCA5, as a traditional and effective method, may become an important model to study its relationship with cancer in the future. Many studies have shown that circRNAs participate in tumor growth and metastasis through many potential mechanisms. Different circRNAs have different pathways and play different roles. This article focuses on retrospective analysis and discussion of multiple studies on the roles of circSMARCA5 in cancer. Many studies have shown that the expression level of circSMARCA5 is significantly related to TNM stage, lymph node metastasis, and other cancer-related indicators –14. In m


Salmonella is a leading cause of bacterial foodborne illness in the world. Although typically associated with foods of animal origin, low-moisture foods, such as tahini, are quickly gaining recognition as an important vehicle of Salmonella exposure. This review offers the Canadian perspective on the issue of Salmonella in tahini and tahini-based products. A summary of several recent food product recalls and foodborne outbreaks related to the presence of Salmonella in tahini and tahini-based products such as halva are presented. The properties of the food vehicles, their production practices, and potential routes of contamination are discussed. Particular focus is placed on the ecology of Salmonella in the tahini production continuum, including its survival characteristics and response to intervention technologies. Salmonella causes 93.8 million illnesses, of which an estimated 80.3 million are foodborne, and 155,000 deaths each year, worldwide [Salmonella are often associated with the consumption of foods of animal origin or fresh produce [w) of 0.85 or below. This category of foods includes chocolate, peanut butter, and tahini, all of which have been linked to multiple large outbreaks of salmonellosis across the globe.Salmonellae are an important cause of human illnesses worldwide. It is estimated that nontyphoidal orldwide . OutbreaSalmonella. Salmonella, however, has the capacity to survive in low-moisture foods for extended periods, up to several years. This prolonged survival increases the risk associated with the consumption of low-moisture foods. As shelf-stable products, a contaminated batch of low-moisture food has the potential to be ingested over time on multiple occasions by multiple people, leading to a wider temporal and geographic distribution of the pathogen than with other food commodities, and a consequent higher number of cases. Three of the largest salmonellosis outbreaks reported in the published literature are linked to low-moisture foods; dried cuttlefish snacks, paprika, and peanut butter with 1505, 1000, and 715 reported cases, respectively [Low-moisture foods are generally regarded as low-risk foods because they cannot support the growth of pathogenic bacteria, including those belonging to the genus ectively ,5,6.Salmonella contamination [In 2004, the World Health Organization’s International Food Safety Authorities Network (INFOSAN) identified sesame-based foods  as an unusual food source for mination . To datemination . OverallSalmonella [Salmonella [In Canada, from 2011–2021, tahini and tahini-based products  have been subject to multiple recalls and updated recalls due to the presence of lmonella . In Canalmonella . In 2019Salmonella in tahini and tahini-based products and provides an overview of the available literature concerning the ecology of Salmonella in sesame seeds, tahini, and halva. Intrinsic properties of sesame seeds, tahini, and halva will be summarized, along with extrinsic factors that can lead to contamination along the processing continuum. Features of the pathogen allowing for long-range survival, mitigation strategies, and areas for further research will also be covered.This review offers the Canadian perspective on the issue of Salmonella consists of two species, S. enterica and S. bongori, that have been subtyped into over 1500 serological variants, or serovars. In terms of human illness, multiple serovars of S. enterica account for over 99% of human salmonellosis cases. Children under 5 years old, the elderly, and people with compromised immunity are most vulnerable to salmonellosis and more likely to develop severe symptoms [The genus symptoms ,35.Salmonella range from 100 to 1011 cells [w) in a food matrix may protect Salmonella from the acidic conditions of the stomach, thus increasing the likelihood of illness from consuming low numbers of the microorganism [Estimates of the infectious dose for 11 cells ,37. Hostorganism ,39. Analorganism ,40,41,42Salmonella ranging from <0.03 MPN/g to 0.46 MPN/g have been found in tahini and tahini-based products linked to salmonellosis outbreaks [Concentrations of utbreaks ,25. BaseSalmonella ser. Brandenburg , Heidelberg (two outbreaks), Mbandaka (three outbreaks), Montevideo (five outbreaks), and Typhimurium (three outbreaks). In 2016 and 2017, an outbreak of a previously undescribed Salmonella serovar, Vari, occurred in five European Union countries . They are used as cooking and baking ingredients. Their high oil content (approximately 50% of the seed’s weight) makes them a valuable source of oil for cooking, cosmetic, and pharmaceutical applications [Sesame seeds are cultivated from the sesame plant  of 0.16 and a pH of 5.9 [Salmonella levels should it be present in any of the raw ingredients.Tahini is the paste produced from ground sesame seeds, which has a high-fat (57–65%) and low-moisture content (<1%) . On averH of 5.9 ,71. TahiH of 5.9 . TraditiH of 5.9 ,72. ThesCountries that are key global exporters of tahini include Lebanon, Syria, Egypt, Greece, and Israel . Tahini Salmonella is present on the initial lot of seeds, this practice can amplify the level of Salmonella by up to 3 logs prior to the roasting and grinding of seeds [Tahini is typically obtained by milling cleaned, dehulled, and roasted sesame seeds ,70. The of seeds .The majority of tahini sold for consumption is made from roasted seeds. Raw tahini, made from unroasted or lightly roasted sesame seeds, represents a small percentage of all tahini sold  ,62. RoasSalmonella levels [Salmonella artificially inoculated onto sesame seeds. The authors noted a period of rapid decline within the first 10 min of roasting, followed by a lower rate of reduction. This first ten minutes of heating corresponded to a reduction in the seeds’ aw, from 0.98 to 0.14. This suggests that during the roasting process, the surviving population of Salmonella cells exhibit an increase in heat resistance as the moisture of the seeds decreases [Studies have demonstrated that the roasting of sesame seeds can reduce a levels ,80. TorlSalmonella, then soaked and dried them to an aw of 0.90. When these seeds were roasted at 130 °C using an air-forced oven, Salmonella populations decreased below the detection limit (1.7 log CFU/g) within 10 min. However, when the sesame seeds had an aw of 0.45 before roasting at the same temperature, the decline in cell populations took longer with an approximate 5-log reduction after 60 min, meaning that 3.5 log CFU/g remained on the seeds after this roasting process [Salmonella present on the sesame seeds depends on the starting aw.Similarly, Zhang and colleagues inoculated sesame seeds with 8.5 log CFU/g of  process . This arSalmonella survives the roasting process, it is likely to remain viable throughout the shelf life of the product. Therefore, tahini and tahini-based products may become contaminated by Salmonella through insufficient or inadequate roasting of contaminated sesame seeds [Salmonella cross-contamination in low-moisture foods has been traced to factors such as poor sanitation practices, poor equipment design, improper maintenance, and poor ingredient control [If me seeds ,80. In a control ,81.Salmonella control practices are available. These documents include the Code of Hygienic Practice for Low-Moisture Foods, CXC 75-2015 [Hazard Analysis and Critical Control Point Generic Models for Some Traditional Foods: a Manual for the Eastern Mediterranean Region [Salmonella in Low-Moisture Foods [Salmonella contamination: prevent ingress or spread of Salmonella in the processing facility, enhance the stringency of hygiene practices and controls in the Primary Salmonella Control Area, apply hygienic design principles to building and equipment design, prevent or minimize growth of Salmonella within the facility, establish a raw materials/ingredients control program, validate control measures to inactivate Salmonella, and establish procedures for verification of Salmonella controls and corrective actions [Guidance documents for manufacturers of tahini and tahini-based products describing  75-2015 ; the Hazn Region ; and there Foods . The Gro actions .Salmonella prevalence levels in tahini from Canada, Germany, and the Middle East ranged from 0.4% to 20%  in tahini. All positive samples were from tahini imported from Middle Eastern countries , even though almost half of the products tested were domestically produced [Salmonella prevalence among the sampled products [Published % to 20%  [13,60,6produced ,63,64. Iproducts . Serovarproducts ,63,64, Tw of 0.18 and pH of 6 [Saponaria officinalis). The syrup is heated to 120–140 °C prior to mixing with the tahini [Salmonella into the product. As an illustration, halva products containing cocoa and/or pistachio demonstrated increased Enterobacteriaceae/coliform/E. coli counts compared to plain halva [Enterobacteriaceae/coliform/E. coli and Salmonella presence, suggesting these counts can be used as indicators of overall process hygiene as opposed to indicators of pathogen presence. Reported Salmonella prevalences in halva ranged from no detections to 11.3%, with Typhimurium DT 104, Poona, and a monophasic strain from serogroup B isolated  is a key determinant for microbial growth as it is a measure of the amount of water available to carry out essential metabolic functions. Salmonella requires a minimal aw of 0.93 to actively grow and reproduce [Salmonella in sesame seeds, tahini, and halva have shown survival periods ranging from at least 4 to 12 months [Salmonella in these foods. Even after prolonged storage, the pathogen remains viable, and therefore, capable of causing disease.Water activity  until exposed to favourable growth conditions, at which point they can resuscitate and resume metabolic and other accessory functions such as pathogenesis.ronments ,82. Uponronments . Among tw conditions. Salmonella cells grown in high NaCl, sucrose, or glycerol form long filaments due to an inhibition of cell septation. Similarly, high osmolarity conditions lead to an alteration of the OmpC/OmpF porin ratios in the outer membrane that lead to altered permeability rates. Permeability can also be affected through increased synthesis of unsaturated fatty acids via FabA [Changes to the cell structure and cell surface have been reported in low-avia FabA . Osmoticvia FabA . Howevervia FabA .Salmonella cells that are tolerant to desiccation undergo a number of physiological changes that may render them less susceptible to other food control measures such as heating, acidification, and/or treatment with biocides [Salmonella cells inoculated onto sesame seeds at a starting aw of 0.45 exhibited a slower rate of decline compared to cells on seeds with an aw ≥ 0.95 when the seeds were roasted at 130 °C [biocides . For exat 130 °C .w food systems and is an area that requires further study.There may be a relationship between biofilm formation and desiccation resistance, since cells that form biofilms have increased resistance to many ecological stresses . Cells tSalmonella to tolerate desiccation varies from strain to strain. Norberto and colleagues reported in a study of 37 Salmonella strains belonging to 16 serovars log reductions ranging from 0.6 to 2.7 when inoculated in dry soybean meal and stored at 25 °C for 18 h [S. enterica serovars Ohio, Montevideo, and Havana, respectively [The ability of for 18 h . There wectively . The natSalmonella. In recent years, there has been intensive research efforts into postproduction lethality treatments of tahini and tahini-based products. The intent is to eliminate Salmonella without compromising the quality properties of the food products. Some of the inactivation treatments tested on sesame seeds, tahini, or halva are presented below.Both thermal and nonthermal treatments of low-moisture foods or their raw materials can be used to control the presence of Salmonella during the manufacturing of tahini, as described in detail in Roasting of the sesame seeds is a key step to control Salmonella ser. Typhimurium in tahini treated at temperatures of 70 °C, 80 °C, and 90 °C. Regardless of the test conditions, the Salmonella cells had a high rate of survival with a maximal reduction of 3 log CFU/mL after one hour at 90 °C. The inactivation profiles demonstrated a rapid decline in the first two minutes of treatment, followed by a tailing curve at all tested temperatures, suggesting the applied thermal treatments were only able to inactivate a subset of the Salmonella population [Some manufacturers may also include a thermal treatment applied to the tahini as a final step . Szpinakpulation .Salmonella [Salmonella serotypes  using a disc diffusion assay method [Salmonella between 2.4 and 2.8 log CFU/mL, whereas 2.0% thyme oil led to a reduction between 2.2 and 3.3 log CFU/mL by 28 days, depending on the storage temperature . However, the addition of essential oil extracts reduced the quality and consumer acceptability of the tahini, as assessed by a sensory panel [Some studies evaluated the impact of incorporating natural antimicrobials into tahini in order to control lmonella ,87,94. TSalmonella ser. Typhimurium by 2.7–4.8 log CFU/mL and 2.5–3.8 log CFU/mL, respectively, in tahini after 28 days at tested temperatures of 10 °C, 21 °C, and 37 °C [Salmonella levels with the use of 0.5% citric acid (0.8 log CFU/g reduction after 7 days at 4 °C) [The antibacterial effects of organic acids, such as acetic and citric acids, have also been tested in tahini. The addition of acetic and citric acids at a concentration of 0.5% reduced nd 37 °C . Similarat 4 °C) .Salmonella spp. in tahini and tahini-based products.Antimicrobial substances, such as metals, chemicals, essential oils, enzymes, and bacteriocins, can also be used in packaging materials . More reSalmonella. Three types of radiation can be used on bulk or packaged products: gamma rays, X-rays, and electron beams [Salmonella ser. Montevideo by gamma irradiation of sesame seeds. Their results showed that the use of 5 kGy was able to reduce Salmonella counts by 5.53 log CFU/g on sesame seeds [Salmonella using gamma irradiation was also tested in tahini [Salmonella, whereas in tahini, an irradiation dose of 2.0 kGy led to a reduction of 4.7 log CFU/g of Salmonella [Ionizing radiation can effectively eliminate microbes that cause foodborne illness, including on beams . D’Oca ame seeds . The inan tahini  and halvn tahini . In halvlmonella ,98. In tlmonella .Salmonella ser. Montevideo on sesame seeds. No synergistic effect of the combined treatments was observed, nor was there any reduction in the quality of the seeds [Two recent studies tested energy-based technologies on sesame seeds and tahini ,100. Xu Salmonella was assessed in tahini. Salmonella reductions ranging from 3.6 to 4.7 log CFU/g were observed after treatment. After two minutes of treatment, the core temperatures of the tahini samples reached, 45 °C, 70 °C, 103 °C, 119 °C, and 144 °C at 220, 330, 440, 550, and 660 W, respectively. The authors noted that microwave heating did not affect acid, peroxide, p-anisidine, or color values of the tahini up to 90 °C [The efficiency of 2450 MHz microwave heating at 220, 330, 440, 550, and 660 W for the inactivation of to 90 °C .Fumigants, such as ethylene oxide and propylene oxide, have shown to be effective for achieving significant reductions in microbial populations in low-moisture foods. More precisely, ethylene oxide is used to treat spices, whereas propylene oxide is used with a variety of foods such as nuts, spices, cocoa beans, and dried fruits .Salmonella contamination [In some countries, including the European Union, the use of ethylene oxide in foods is not permitted. In recent years, multiple recalls of sesame seed products due to the presence of ethylene oxide residues occurred in the European Union. It is believed that ethylene oxide was used for fumigating sesame seeds to eradicate mination .2) to reduce Salmonella levels on sesame seeds. A treatment with 500 mg ClO2/kg reduced the number of Salmonella from 7.6 log CFU/g to 4.9 log CFU/g on sesame seeds [Golden and colleagues tested the use of chlorine dioxide , or adopt a filamentous shape [Salmonella [Salmonella such as PCR should be used in conjunction with a primary enrichment step [There are many challenges associated with the detection of ratories . Furtherus shape ,106. Prilmonella ,108. Rapent step . The reqent step .Salmonella and tahini were identified. Research directed in these areas can provide information required to mitigate the public health risks associated with the presence of Salmonella in tahini and tahini-based products:Salmonella serovars associated with sesame seeds, tahini, and tahini-based products;Understanding the ecology and potential virulence of Knowledge of the response of sesame and tahini-associated serovars, and whether findings from different serovars can be generalized to the genus;Salmonella in tahini and other colloidal foods, particularly the impact of rheological and physical properties ;Survival of Salmonella adaptations to high-fat, low-moisture foods such as tahini  and the potential impact on detection and lethality treatments;The impact of interventions, alone and in combination, throughout the tahini manufacturing process, particularly at the seed soaking and roasting steps;Salmonella response in commercial settings to typical sesame seed roasting parameters, in order to understand the physiology of cells that survive the roasting process;Improved traceability and environmental/process monitoring of sesame seeds, tahini, and tahini-based products to allow for a more detailed root cause analysis of food safety incidents;Salmonella growth dynamics during primary enrichment to improve detection and enumeration methods, with an emphasis on reducing the time to result.In the course of preparing this review, the following knowledge gaps concerning Salmonella in tahini and tahini-based products is one of global concern. The tahini production continuum from sesame seed to consumption is one that transcends multiple borders, and requires international cooperation for effective management, trace-back, and source attribution. Several features of the Salmonella–tahini relationship make it an intractable food–pathogen combination as demonstrated by the spate of recent food safety incidents associated with it. The long shelf life and the ready-to-eat nature of tahini make it an ideal vehicle for a long and sustained exposure to Salmonella. Further, the high fat, low-moisture environment of tahini improves Salmonella survival and resiliency to a variety of lethality treatments. Given the low infectious dose of Salmonella, complete elimination from a batch of contaminated tahini would be required to prevent illness, a goal that is difficult to achieve in this food matrix.The issue of Salmonella to safe levels in tahini underscores the importance of preventing contamination. For many tahini producers, roasting is the only lethal step applied in the manufacturing process. A validated roasting procedure with strict ingredient and parameter control is key. Similarly, given the high potential for contamination after roasting, process verification and environmental monitoring of the production facility is important. Correlations between total aerobic mesophile/Enterobacteriaceae/coliform/E. coli counts and Salmonella presence have not been observed, but enumeration of the former groups can provide information on the hygiene of the production process. Verification and monitoring of the entire production continuum, from sesame seed production to tahini distribution, may not be possible given the reliance on imported products and the complexity of supply chains. This complexity also complicates food safety investigations and root cause analyses in the event of food safety incidents. Therefore, it is important for the implicated countries to adhere to a set of internationally agreed-upon standards and procedures to ensure the safe manufacture and consumption of this global food product.The difficulty in eliminating


Egypt is the country with the highest known hepatitis C virus (HCV) prevalence worldwide. The origin of gender differences in HCV prevalence is not usually well understood. This systematic review and meta-analysis aimed to review and evaluate the gender differences in HCV infection rates amongst Egyptians. Such data would be important to support prevention and control programs aiming to minimize HCV-related morbidity and mortality. PubMed, Scopus, and Web of Science (WOS) were searched for relevant articles published from 1st January 2011 to 13th December 2021, using the search terms (HCV OR “hepatitis C” OR hepacivirus) AND . At first, retrieved articles were screened, and then relevant data were extracted and analyzed. Descriptive statistics were used for data analysis. Out of 616 studies from databases, only 30 were included after the full-text screening, with 193,621 included participants: 97,597 male and 96,024 female. The overall seroprevalence of HCV antibodies in all included studies was 0.02 (CI − 0.23 to 0.28), with no significant difference between males and females. However, HCV RNA positivity was significantly more prevalent in males than females in adults and the general population (after excluding high-risk groups). In children, no statistically significant differences between males and females were found in the seroprevalence of HCV antibodies nor in the prevalence of PCR positivity. HCV RNA positivity is significantly higher in males than females in adults, while there are no gender differences in children. Globally, 1.5 (1.3–1.8) million people are newly infected with HCV every year, and 58 (46–76) million people are living with chronic HCV infection, with a global prevalence of 0.8% (0.6–1.0%) in the general population. The highest prevalence in the Eastern Mediterranean Region is 1.6% (1.4–1.8%), 290,000  people die from hepatitis C-related causes every year, and only 21% of people are diagnosed with HCV infection, and 62% of them receive treatment2.Gender is an essential determinant of social outcomes, including health. Research has shown a growing interest in health-related gender differences and raises the question of gender-biased differential response that is relevant in many health fields, including the prevalence, risk factors, clinical features, and treatment of diseases. Still, the epidemiological pattern of HCV infection in research and medical practice requires further knowledge of the potential role of gender differences. Thus, assessing gender-level change in HCV prevalence may help identify population subgroups most likely to suffer an increased infection rate, thus enabling health authorities to plan targeted interventions for these changes6, while others support the view that HCV infection appears to be prevalent and progresses more rapidly in males than in females11. On the other hand, some studies found gender differences are artifactual, with nearly flat rates of HCV infection15. Also, some studies found a decreased rate of liver cirrhosis and hepatocellular carcinoma (HCC) in females16, more progress to hepatic fibrosis in males, and more liability to adverse events of direct-acting antivirals (DAAs) in females17.The origin of gender differences in HCV prevalence is not well understood, and some hypotheses tried to explain this difference. The salience of gender in positioning women at increased risk of exposure to HCV infection has been confirmed by some studies18. On the other hand, male predominance can be explained by differences in daily life conditions, environmental experiences, and social, cultural, and occupational aspects taken up more frequently by men than women, especially in marginalized groups and slum areas such as IV drug use, circumcision, shared use of toothbrushes or shaving razors, tattooing, wet cupping (Higama), or illegal sexual intercourse, in addition to blood transfusion emergencies19. The hormonal hypothesis and other mechanisms have been invoked, such as cellular mosaicism, genes escaping X chromosome inactivation, skewed X chromosome inactivation, and miRNAs encoded on the X chromosome20. In addition, gender differences in HCV infection rates might reflect differences in the patterns of gender-specific risky behaviors22.Compared to men, women are more exposed to syringes, blood, and blood products, especially during pregnancy and labor, and ear piercing, and thus run a higher risk of HCV infection. Biological sex with female predominance has been associated with differences in rates of spontaneous HCV clearance, with a possible role of sex hormones in determining host susceptibility to viral infections23.Historically, Egypt is one of the world countries with the highest prevalence of HCV infection. Over the past decade, Egypt has continued efforts to achieve HCV control and works towards the common goal, targeted by the WHO, of the elimination of viral hepatitis by 2030. The universal access to treatment with the introduction of DAAs has resulted in a paradigm shift in HCV management and declining mortality. A large Egyptian study showed a marked decrease in mortality in Egyptp < 0.001)10, 9, and 8. While other studies found more anti-HCV prevalence in women than in men in special situations:  among family contacts of HCV-positive children24,  in patients with coronary heart disease5, and  in apparently healthy blood donors3.In Egypt, some studies indicated that anti-HCV prevalence in the general population was higher in males than in females  for relevant articles published from 1st January 2011 to 13th December 2021. Our protocol was registered to the International Prospective Register of Systematic Reviews .We searched the target databases one by one using the following search terms: (HCV OR “hepatitis C” OR hepacivirus) AND .Any Egyptian observational cross-sectional study containing original information regarding the prevalence of HCV on both males and females published in the English language from 2011 or later till the end of 2021 was included irrespective of the governorate, studied group, and age of participants. Studies that did not specify gender, non-Egyptian studies, studies with a mixed population with no definite data for Egyptians, studies in which prevalence was not the primary concern, editorials, reviews, abstracts, posters, commentaries, and non-human studies were excluded.MO, YA, HN, FA), and a discussion with the research team solved any disagreements. The same four independent reviewers did full-text screening for articles selected in the previous step. Data extraction for finally included studies retrieved from the full-text screening was done independently. Relevant data were extracted to a pre-prepared excel file. Quality assessment was done by two independent reviewers . We used The Joanna Briggs Institute (JBI)25 tool for the quality assessment of prevalence studies.Title and abstract screening were done by four independent reviewers  was used with a random effect model to compare males and females. A funnel plot was used to check for publication bias using egger's test  . Heterogeneity was assessed using the I44 were finally included in our meta-analysis after full-text screening  as shown in Fig. The seroprevalence of HCV antibodies among the Egyptian population using serum antibodies test by ELISA, based on the results of the random-effects method, there were no significant differences between male and female seroprevalence of HCV. The overall seroprevalence was 0.02 , and the heterogeneity was high  as shown in Fig. Eight studies used PCR to measure the prevalence of HCV; based on the results of the random-effects method of these studies, there was a statistically significant increase in the prevalence of HCV RNA positivity in males than females  and the heterogeneity was high  , and the heterogeneity was high  after excluding studies on high-risk groups, males showed a significantly increased prevalence of both HCV antibodies and HCV PCR positivity .Egypt is the country with the highest HCV prevalence worldwide. Six percent of individuals aged 1–59 years had a positive result on the hepatitis C antibody test, and 4% were found to have an active infection. The prevalence of hepatitis C was higher among men than women in most age groups46. The worldwide pooled seroprevalence of HCV in children in these studies was low, < 1%47. In 2015, the Egyptian Demographic and Health Surveys (EDHS) tested 10,044 children  to estimate HCV prevalence in those aged 1–14. The results showed that HCV prevalence in the group aged < 15 years was 0.4%. HCV antibody seroprevalence was 0.7% with viremia of 0.2% in male children, while HCV antibody was seroprevalent in 0.2% with the presence of viremia in 0.1% of female children10. The current meta-analysis shows no significant differences between male and female children in HCV antibody prevalence or viremia.There is limited published data on gender-based differences in children48. Similarly, the results of this meta-analysis showed that the prevalence of HCV viremia was significantly higher in males than females in adults. This difference may be attributed to males being more affected by schistosomiasis disease burden and hence were the main target of the parenteral antischistosomal therapy (PAT) campaign49 with high risk for parenteral virus transmission, including HCV. Also, the lifestyle of males makes them more exposed to various risk factors for HCV transmission50.The EDHS reported that seroprevalence of HCV antibodies was 14.7% among the adult population aged 15–59 years at 14.7% with a national viremic prevalence of 9.7%, which was higher in males than in females in all studied age groups in 200851. Total HCV seroprevalence in the age groups 15–59 years was 10%, with a viremic prevalence of 7%. HCV seroprevalence in female adults was 8.1%, with viremia of 5.5%, while HCV seroprevalence in male adults was 12.4%, with a viremic prevalence of 9.8%8.In 2015, there was significantly lower HCV prevalence in those aged 15–19 years compared to the 2008 data (14%), and this points to a significant decrease in new infections in the age groups 15–19 yearsThere were not enough data from the included studies to determine the sources of heterogeneity. It may be due to differences in populations, regions, geographical locations, seasons, settings , and used screening methods. Unmeasured covariates  could have contributed to variability in outcome estimates. Gender differences among high-risk populations  were not considered in this study to avoid overestimating the problem.HCV RNA positivity is significantly higher in males than females in adults, while there are no gender differences in children.Supplementary Figures.


Data on alkaloid interactions with the physiologically important transition metals, iron and copper, are mostly lacking in the literature. However, these interactions can have important consequences in the treatment of both Alzheimer’s disease and cancer. As isoquinoline alkaloids include galanthamine, an approved drug for Alzheimer’s disease, as well as some potentially useful compounds with cytostatic potential, 28 members from this category of alkaloids were selected for a complex screening of interactions with iron and copper at four pathophysiologically relevant pH and in non-buffered conditions (dimethyl sulfoxide) by spectrophotometric methods in vitro. With the exception of the salts, all the alkaloids were able to chelate ferrous and ferric ions in non-buffered conditions, but only five of them  evoked some significant chelation at pH 7.5 and only the first two were also active at pH 6.8. By contrast, none of the tested alkaloids chelated cuprous or cupric ions. All the alkaloids, with the exception of the protopines, significantly reduced the ferric and cupric ions, with stronger effects on the latter. These effects were mostly dependent on the number of free aromatic hydroxyls, but not other hydroxyl groups. The most potent reductant was boldine. As most of the alkaloids chelated and reduced the ferric ions, additional experimental studies are needed to elucidate the biological relevance of these results, as chelation is expected to block reactive oxygen species formation, while reduction could have the opposite effect. Although iron and copper have a number of physiological functions in the human body, their dyshomeostasis has been observed in several diseases, including cardiovascular diseases, cancer and some neurodegenerative disorders, including Alzheimer’s disease ,2,3. TheNotwithstanding that many papers reported the antioxidant effect of alkaloids , the datFor this reason, the aim of the study was to assess the interaction of 28 isoquinoline alkaloids  with iroIn the first step, all the alkaloids were screened for their ability to chelate iron ions in a non-buffered environment using the ferrozine spectrophotometric method, which is based on competition between a tested alkaloid and ferrozine for free (non-chelated) iron . All theIn the next step, chelation was tested in buffered conditions at four pathophysiologically relevant pHs, ranging from 4.5 to 7.5. In general and in contrast to the non-buffered conditions, only 5 alkaloids, galanthine, glaucine, corydine, corydaline and tetrahydropalmatine, evoked significant chelation at pH 7.5 at a rate of 10 times that of the ferrous ions. However, only two, galanthine and glaucine, were also active at pH 6.8. Galanthine A was theIn order to verify whether the most active alkaloid glaucine could also chelate the ferric ions, a non-competitive approach was used. Firstly, the spectra of glaucine and its mixture with ferric ions were compared. Under all the tested pH, as well as in the non-buffered conditions, the addition of ferric ions clearly shifted one peak in the spectra of the glaucine, suggesting a complex formation . The stoAfter determining the iron-chelation activity, the copper chelation was tested by two competitive methods, with hematoxylin and bathocuproinedisulfonic acid disodium salt (BCS). However, no cupric or cuprous chelation was observed in either of these assays, in contrast to the clinically used copper chelator, trientine (data not shown).The ferrozine and BCS methods were also used for the determination of the metal reduction activity of the alkaloids. With the exception of the protopine alkaloids, all the other tested compounds were shown to be active reductants, although their potency was very different. Their effect was especially strong in the case of the copper ions .The iron was reduced under both non-buffered conditions, as well as at pH 4.5. With the exception of a very mild ferric reduction by glaucine at pH 5.5, none of the tested alkaloids were able to reduce the ferric ions at pH 5.5 or under higher-pH conditions. There were also apparent differences between the ferric reduction at pH 4.5 and in the unbuffered conditions. In line with the chelation data, the reduction at pH 4.5 was more pronounced than in the non-buffered conditions. In the formed case, the reduction activity increased with the increasing alkaloid:iron ratio B. The moIn the unbuffered conditions, there were six alkaloids with bell-shaped reduction curves. Four aporphine alkaloids , sinoacutine and scoulerine achieved the highest reduction, at ratios of approximately 1:1 to 2:1, while there was a decrease in the reduction activity at higher ratios , with the exception of glaucine under some conditions. The bulbocapnine and boldine were even more potent and achieved complete reduction at ratios of 1:10 and 1:50, alkaloid:Cuectively A–C. Amonectively  is shownortho-dimethoxy group had mostly similar potencies to berberine . The tested Amaryllidaceae alkaloids were not structurally homogenous, but the results confirmed and extended previous findings  can chelate iron and their chelation properties can be fairly strong  many alkaloids were not supposed to possess metal chelation activity based on their structure ; and  and this could also have had an impact as, for example, the reduction of ferric ions is markedly influenced by the selected solvent . There i4 ∙ 5H2O), cuprous chloride (CuCl), DMSO, ferric chloride hexahydrate (FeCl3 ∙ 6H2O), ferrous sulfate heptahydrate (FeSO4 ∙ 7H2O), 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-4′,4′′-disulfonic acid sodium salt (ferrozine), hydroxylamine hydrochloride (hydroxylamine), hematoxylin, hydrochloric acid (HCl), BCS, sodium chloride (NaCl), sodium acetate, acetic acid, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), HEPES sodium salt and trientine were purchased from Sigma-Aldrich (Czech Republic), while deferoxamine was from Novartis (Switzerland), boldine was from Carl Roth (Germany), glaucine was from Cayman Chemical  and galanthamine hydrobromide was from Changsha Organic Herb . Chlidanthine and galanthamine were isolated from Chlidanthus fragrans herb [Zephyranthes robusta (Herb. ex Sweet) [Corydalis cava (L.) [Eschscholia californica [Argemona platyceras [Fumaria officinalis L. [Berberine Cl, isocorydine HCl, cupric sulfate pentahydrate  , bulbocaava (L.) , califorifornica , platyceatyceras  and sinanalis L. .All experiments were carried out on SynergyTM 2 Multi-Mode Microplate Reader  in 96-well microplates with the use of methodologies described previously ,36. Defe3 ∙ 6H2O), ferrous (FeSO4 ∙ 7H2O) and cupric (CuSO4 ∙5H2O) ions were prepared in distilled water , as well as hydroxylamine, ferrozine and BCS solutions. The stock solution of cuprous (CuCl) ions was prepared by dissolution in an aqueous solution of 0.1 M HCl and 1 M NaCl. The solutions of tested alkaloids and hematoxylin were prepared in DMSO. All experiments were performed under non-buffered and buffered conditions .The stock solutions of ferric  and 5 min later. All measurements were performed at least in duplicates. Deferoxamine was used as a positive control.The same methodology was used for determination of ferric ion reduction. Hydroxylamine was used as a positive control, causing 100% ferric-ion reduction.The alkaloid DMSO solutions were mixed with various buffers and cupric-ion solution and after 2 min of shaking, hematoxylin or DMSO (blank sample) was added. The absorbance was measured after 3 min of shaking and later after additional 4 min at different wavelengths, depending on the tested pH, as reported previously .This assay was identical to the ferrozine assay, except for the use of the BCS reagent specific to cuprous ions. Briefly, alkaloid solutions were mixed with cupric or cuprous solutions and incubated for 2 min with or without buffer. Hydroxylamine was added prior to cuprous ions to keep them in reduced form or after the mixing with cupric ions to reduce non-chelated cupric ions to cuprous ions. The amount of non-chelated copper was determined spectrophotometrically after the addition of the BCS reagent.As in the case of the ferrozine method, this method is also suitable for determining the copper-reducing capabilities of test substances. Again, hydroxylamine was employed for complete reduction of cupric ions.The stoichiometry was assessed according to the previously published methodology . The metThe concentrations of both components  changed, while their concentration sum was kept constant. This was based on the principle that the highest quantity of the complex is formed under ideal conditions, which implies that under such conditions, the maximal absorbance is observed at the concentration ratio corresponding to the stoichiometry of the complex. The molar concentrations of alkaloid + ferric ions were kept constant at 100 μM, while their molar concentration ratios were continuously changed from 1:4 to 6:1. The measurements were performed against a blank composed of the buffer and methanol at a ratio of 2:1.For the complementary approach, the molar concentration of alkaloid was continuously changed, while the concentration of iron was kept constant throughout the series of samples (10 μM) with different molar concentration ratios, ranging from 1:4 to 6:1 . The blank composition was the same as that used in the Job’s method. Based on the determined molar absorption coefficients and measured spectra, assessment of the stoichiometry was performed with construction of theoretical lines mimicking the absorbance of the most probable stoichiometries  and its corresponding blank (without indicator) divided by the difference between the control sample (the known amount of iron/copper ions without the tested substance) and its control blank.y means the chelation efficiency in %, x is the common logarithm of concentration ratio of alkaloid:iron/copper and k is the slope of the curve.Normalized dose-dependent curves were constructed by use of GraphPad Prism version 7.03 for Windows by GraphPad Software  for each condition from at least five points . The efficiency of chelation at concentration ratios of 10:1 and 1:1  was calculated from the curve equation:t-test using MS Excel. Reduction lines were constructed by linear regression model from points, in which the reduction was significant vs. the solvent and the maximal reduction caused by the hydroxylamine. The 95% confidence intervals were created as well by GraphPad Software.Data are expressed as means ±SD. Results for individual alkaloids were compared with the standard chelators and the solvent by one-way ANOVA test followed by Bonferroni’s Multiple Comparison Test. Significant difference in reduction activity vs. the solvent (DMSO) was evaluated by ortho hydroxymethoxy site, a methylendioxy ring, or even an ortho dimethoxygroup are able to chelate ferrous or ferric ions under non-buffered conditions when DMSO is used as the solvent. However, most alkaloids lose their chelation effect in buffers and, in particular, with decreasing pH. At the same time, with the exception of protopines, most alkaloids are fairly potent reduction agents, particularly for cupric ions, at all the pathophysiologically relevant pH tested in this study (4.5–7.5). This paper opens new fields in this research as it is well known that metal chelation is or can be suitable in several pathological conditions, e.g., in cardiovascular disease and Alzheimer’s disease. On the other hand, metal reduction can potentiate the metal-based Fenton reaction, and this could render these compounds unsuitable in the treatment of these diseases but suitable for cancer treatment. Nevertheless, in compounds with chelation, reduction and, possibly, antioxidant effects, experimental data are crucial to define the final anti- or prooxidant effects.This study confirmed previous isolated results on the chelation effects of alkaloids. In particular, isoquinoline alkaloids with an


The rate of planned pregnancies was 76.8%, and the subfertility rate was 4.9%. Disease activity improved every trimester, and approximately 20% experienced an improvement in the second trimester. Planned pregnancies and corticosteroids use (≤10 mg daily) were protective factors for APO in RA pregnancies . There was no significant association between APO and disease activity or DMARDs used before and during pregnancy. Regarding the comparison between the RA group and the controls, RA mothers were significantly older (p = 0.001), had shorter pregnancies (p < 0.001), and had neonates with a lower birth weight (p < 0.001).Women with rheumatoid arthritis (RA) may carry an increased risk of adverse pregnancy outcomes (APO). The aims of this study were to compare pregnancy outcomes in RA patients as compared to the general obstetric population (GOP) and to identify a risk profile in RA. A case-control study was conducted on 82 prospectively followed pregnancies in RA and 299 pregnancies from the GOP. The mean age at conception was 31.50 ± 4.5 years, with a mean disease duration of 8.96 ± 6.3 years. The frequency of APO in RA patients was 41.5%, 18.3% experienced spontaneous abortions, 11.0% underwent preterm deliveries, 7.3% had small for gestational age infants, 4.9% experienced intrauterine growth restriction, 1.2% experienced stillbirth, and 1.2% suffered from eclampsia. The risk of APO was correlated with a maternal age higher than 35 years ( Rheumatoid arthritis (RA) is a chronic, autoimmune, inflammatory disease affecting the joints and musculoskeletal structures. Indeed, RA can induce progressive disability if not treated properly. The prevalence of this disease in European countries varies from 0.31% to 0.9% of the population, with a man:woman ratio of 1:3 . The conSince the beginning of the 1990s, the link between estrogen levels and the severity of induced arthritis has been demonstrated ,7. PregnAutoimmune diseases represent a risk factor for adverse pregnancy outcomes, as well as autoimmune disease activity. The more severe type of RA  is indicative of higher active RA, more extensive joint destruction, and extraarticular manifestation. Study population reports have shown a correlation between RA, disease activity, and pregnancy complications, such as premature rupture of membranes, low birth weight, and intrauterine growth restriction, stillbirths, eclampsia, and increased risk of caesarean section ,13,14.Pregnancy outcomes can be severely influenced by disease activity and immunosuppressive treatment during the preconception period and gestation. An inactive disease before conception usually remains stable during pregnancy . AnotherIn this paper, we aimed to describe the adverse pregnancy outcomes (APO) among pregnancies in RA patients and disease course during conception for these women, identifying characteristics that were associated with APO. As a secondary objective, we compared the frequency of APO in the RA cohort to that of a control group.An observational, multicentered case-control study of RA patients who became pregnant was conducted, specifically at the Rheumatology Department of the Dr. I. Cantacuzino Hospital in Bucharest, but also in four other clinics across the country, namely at the Department of Internal Medicine and Rheumatology, Dr I. Cantacuzino Clinical Hospital and Department of Internal Medicine and Rheumatology, “Sfanta Maria” Hospital from Bucharest, Rheumatology Department, University of Medicine and Pharmacy of Craiova, Rheumatology Division, 2nd Rehabilitation Department, Rehabilitation Clinical Hospital Cluj-Napoca, and the Rheumatology Department, Grigore T. Popa University of Medicine and Pharmacy, Iasi. Inclusion criteria were women older than 18 years old at conception, with an RA diagnosis before pregnancy, who became pregnant between 2012–2022. The exclusion criterion was voluntary termination of pregnancy for either medical or personal reasons. Although the inclusion criteria were fulfilled for 98 cases, in the final study, 82 pregnancies were included in the case group, since 16 pregnancies (16.3%) were excluded due to the mother’s choice to have an abortion. The study included 82 pregnancies from 59 women with RA, as follows: 1 patient had 4 pregnancies, 4 patients had 3 pregnancies, and 12 patients had 2 each, with the rest having only 1 pregnancy. Due to the chronic disease’s heterogeneity, which presented itself differently during conception, we treated each pregnancy as a distinct case. The local ethics committee gave its approval (Study Code 5981/8 April 2022), and written consent for all patients was obtained. Patients were examined by a rheumatologist and obstetrician 3 months before conception, throughout pregnancy, and 3 months post-partum. At every visit, data regarding disease activity measured by DAS28CRP , rheumatoid arthritis therapy, and pregnancy evolution and outcomes were collected. Based on pregnancy outcomes, we divided the subjects into two subgroups, comparing disease aspects between them, defined as adverse pregnancy outcome (APO) and no adverse pregnancy outcome (No APO). The APO group included preterm delivery (before 37 weeks of gestation), small for gestational-age infant (SGA), intrauterine growth restriction (IUGR), eclampsia, spontaneous abortion, and stillbirth. A fetus was defined small for gestational age when its estimated fetal weight or abdominal circumference  was below the 10th percentile for the gestational age reference ranges. Abnormal fetal Doppler evaluation associated with estimated fetal weight or abdominal circumference below the 10th percentile defined fetuses with intrauterine growth restriction ,20. EclaCase-control patients were 299 consecutive women over 14 years old admitted to Filantropia Clinical Hospital in 2014 for delivery. The data from the control group were collected in 2014, within research grant 20062, from Carol Davila University of Medicine ad Pharmacy.Demographic variables were collected for cases and control patients enrolled. For RA patients, we collected variables 3 months before conception, throughout pregnancy, and 3 months post-partum, such as disease activity measured by DAS28CRP , rheumatoid arthritis therapy, pregnancy evolution and outcomes, as well as the immunological risk factors and the prevalence of other autoimmune diseases as secondary Sjögren’s syndrome, Hashimoto’s thyroiditis, and antiphospholipid syndrome. In order to define a planned pregnancy, the patients underwent a pregnancy planning consultation. The rheumatologist advised the best RA medication compatible with pregnancy, limiting the NSAIDs intake in periovulatory period, testing for antiphospholipid antibodies and anti-RO antibodies, as well as postponing the procreation process for women with an active disease.Disease activity was measured using DAS28(CRP) 3 months before pregnancy and at every trimester during pregnancy. A DAS28(CRP) value below or equal to 3.2 (remission and low disease activity) was considered as inactive disease, whereas a value greater than 3.2 (moderate and high disease activity) was defined as an active RA. A RA flare was defined as progressing to a more advanced disease stage based on the DAS28(CRP), considering a more active disease. The number of flares during pregnancy and 3 months postpartum were registered.The information collected on the management of RA included the type of medication, either biological DMARDs or synthetic DMARDs, 3 months prior to conception, and in each trimester.t-test and the Mann–Whitney U test with a two-tailed hypothesis for continuous data, and the Pearson Chi-square and Fisher’s exact test for categorical data, respectively. Having more than one pregnancy per patient, for odds rations (OR) and 95% confidence interval (CI), we used generalized estimating equation analyses adjusted for age. For all tests, a significant statistical result was considered at p-value < 0.05.The system used for descriptive and analytical statistics was IBM SPSS Statistics version 20.0. Values of categorical variables were registered as numbers and percentages while continuous variables were presented as mean and standard deviation (SD), median, and minimum and maximum, respectively, depending on the normal distribution. To determine any significant associations, the data were analyzed using Student’s The RA cohort’s characteristics are illustrated in The RA seropositivity rate (rheumatoid factor and/or ACPA positivity) was very high in our group, with a value of 86.6%. Out of the 37 patients who were tested for the auto-antibodies anti-Ro/SSA and anti-La/SSB, 3 were positive, and they were closely monitored for congenital heart block throughout the pregnancy, with only one pregnancy presenting a spontaneous abortion.The antiphospholipid antibodies were found in 10% of cases (4 patients from the 40 examined), and all of them followed individualized treatment with low-dose aspirin in monotherapy or associated with heparin therapy according to the latest EULAR recommendations , taking Thirty-six patients (54.5%) gave birth by cesarean section. The mean neonate’s birth weight was 2961.8 ± 508.4 g; low birth weight was found in 15.5% of cases.During pregnancy, only 14.6% of RA patients had a flare, but 72.7% suffered disease relapse in the first 3 months post-partum.Analyzing the RA activity using DAS28(CRP), an inactive disease was more common before pregnancy, as well as during each trimester. Moreover, when compared to the preconception period, disease activity improved every trimester by a maximum of roughly 20% in the second trimester.The case group included 82 pregnancies from 59 patients diagnosed with RA, of which 80.5% (N = 66) had a live birth, 18.3% (N = 15) had a spontaneous abortion, and 1.2% (N = 1) had a case of stillbirth . Based oOne of the premature infants was diagnosed with a congenital cardiac malformation, namely a bicuspid aortic valve, with normal heart function, and also with a congenital nasal dermoid cyst. In this particular case, the RA mother experienced rheumatoid vasculitis and was treated with rituximab for 5 years, with the last infusion being one year before conception, with no specific anti-rheumatic drugs taken during the preconception period or during the pregnancy.There was only one case of eclamsia in the third trimester, in a 31-year-old primiparous, non-obese woman with spontaneous pregnancy. The patient had a RA therapeutic regime compatible with pregnancy, treated with low-dose aspirin prophylaxis for a hereditary thrombophilia with a low risk of thrombosis, levothyroxine for Hashimoto’s disease and sulfasalazine 2 g daily for RA control, with an inactive disease under this medication during pregnancy. She was diagnosed with gestational diabetes at 28 weeks of gestation and high blood pressure, proteinuria, edema, and seizures recognized as eclampsia at 38 weeks of gestation. The patient gave birth by cesarean section to a 3090 g healthy infant, with an APGAR score of 8/10.In our cohort, only one woman needed assisted reproductive technology, at 35 years old with a good pregnancy outcome. She was diagnosed with infertility 5 years before RA diagnosis; she had an inactive rheumatic disease before conceptions and during pregnancy, being under treatment with 10 mg of oral prednisone, until 12 weeks of gestations, without immunosuppressive therapy.During the 3 months before conception, 32.9% were without any RA treatment, the biologic DMARDs being used preferentially, in 17.1% of women . The bDMIn contrast, during pregnancy , the treRegarding the biological therapy during pregnancy, 17.1% (N = 14) were treated with anti-TNF inhibitors and rituximab, with eight of them discontinuing the therapy at a positive pregnancy test result, at around 4 weeks of gestation. Five subjects required biological treatment until the second trimester (four with etanercept and one with infliximab). There was one case of certolizumab initiation in the second trimester due to insufficient control of the disease activity. This patient had a high disease activity in the preconception period and in the first trimester, being under oral prednisone (10 mg daily). Ultimately, she had a preterm delivery at 34 weeks of gestation, with an IUGR fetus, the birth weight of the neonate being 1800 g.The only case of stillbirth at 21 weeks of gestation was an unplanned pregnancy in an obese patient, who received MTX and infliximab in the first trimester (with an inactive disease before and during gestation), without any other associated conditions.p = 0.028, OR (95%CI) = 5.59 (1.53–23.08)). An important finding was the fact that planned pregnancies were statistically significantly associated with no adverse pregnancy outcome  = 0.12 (0.03–0.43)). Hence, the duration from decision-making to conception was shorter for the APO group, with more than half of the subjects having a median time to pregnancy of 0, with a borderline signification (p = 0.052), but this result could be biased due to the high percentage of unplanned pregnancies in the APO subgroup.The two subgroups were mostly similar in terms of demographic features. Even though there was no significant difference in the average age at conception, women older than 35 years were more likely to have an adverse pregnancy outcome = 1.01 (1.00–1.02)). All 10 neonates with LBW were from mothers with pregnancy complications, as follows: three preterm deliveries, three small for gestational age infants, and four with IUGR. Evaluating the birth weight while comparing the planned and unplanned pregnancies, a significantly higher percentage of neonates with LBW had RA mothers with unplanned pregnancies .For the 66 live births, infants from the APO subgroup had a significantly lower birth weight . On the other hand, there was no association between GC therapy and low birth weight (p = 0.715).Furthermore, we analyzed the impact of the disease activity before delivery on birth weight, comparing the infants ≤2500 g and those with a weight >2500 g by mother’s disease activity in the third trimester. The results showed that only 3 cases out of 10 with LBW infants had an active condition in the third trimester  of them had an active disease before conception, and 8 (53.3%) had an active disease during the first trimester.p = 0.016, OR (95%CI) = 0.19 (0.04–0.94)). There were five patients treated with MTX during the first trimester of pregnancy, and three of them had a spontaneous abortion and one had a stillbirth. The fifth had no adverse outcomes.The treatment administered 3 months before and during pregnancy was compared between the two cohorts and the results showed no difference regarding the use of disease-modifying anti-rheumatic drugs, neither synthetic nor biological. A significant result was regarding the intake of orally corticosteroids during pregnancy, namely prednisone ≤10 mg daily. This increased the chance of having a live birth with no maternal or fetal complications (p = 0.001). The frequency of APO is higher for RA patients (27.3% vs. 19.1%), without a significant p-value. Here, C-section delivery was preferred in the RA group compared to the controls, without statistical significance. The neonates from RA mothers had a lower birth weight compared to controls , with LBW being more frequent in the RA cohort, with borderline p-values, p = 0.064. Furthermore, pregnancy length is significantly shorter for RA patients, with premature deliveries being more common (13.6% vs. 8.7%); however, there is no statistical significance. In terms of adverse pregnancy outcomes, SGA was slightly more prevalent in the RA group, while IUGR was more frequent among controls but not statistically significant . There were no cases of eclampsia or pre-eclampsia found in the control group.Regarding the control group, there were five twin pregnancies; thus, we included 304 neonates in the control group. When comparing the 66 births registered in RA patients with a control group  the resuHerein, we described a sample of 82 pregnancies in patients diagnosed before conception with RA, analyzing the potential factors that were associated with adverse pregnancy outcomes. Our results showed a delay in family planning when it comes to those women suffering from a chronic disease, such as RA, concordant to other authors . MoreoveAccording to our findings and in line with the literature, a carefully planned pregnancy, taking into account disease activity and, in particular, potential risks for malformations induced by RA medication, is critical for enhancing the likelihood of avoiding adverse pregnancy outcomes ,26.In our study, 16.3% of women required a voluntary termination of pregnancy, while in a Romanian statistical analysis from 2019, the rate of fertile women who required this procedure was 10.9‰ .Our results showed that miscarriages, preterm deliveries, small for gestational age infants, and intrauterine growth restriction were the most common complications, findings which are in accordance with other studies ,29. MoreA more specific entity was that one mother in our population developed eclampsia at 38 weeks of gestation, a very rare condition defined as high blood pressure, proteinuria, edema, and seizures. To our knowledge, this is one of the few cases of eclampsia in a rheumatoid arthritis pregnancy described in the literature.The rate of cesarean section for RA group was 54.4%, the frequency being higher as compared to the control group. This result is in accordance with other studies ,30.Miscarriage had the largest percentage when analyzing the adverse pregnancy outcomes. Moreover, out of the 15 cases of spontaneous abortion, more than half did not benefit from medical preconception counselling. Consequently, in some of the cases, the RA medication before conception was not in accordance with the recommendation at that time. A further significant result in this paper was the association between unplanned pregnancy and lower weight at birth.In our study, only 14.6% of patients had a flare during pregnancy, and it did not have a great impact on the pregnancy outcome, results that are very different from the findings of a previous study of 46 RA pregnancies between 2001 and 2009, in which more than half of the subjects had at least one flare . ConcernThe risk factors for disease relapse after birth is a controversial topic, and there is not sufficient data to establish exactly where the disease activity during pregnancy or if the type of treatment has a definite effect on post-partum flares ,32.Even though pregnancy can improve RA, one of the most significant factors to consider when planning a pregnancy is disease activity, which can be linked to both a long time to conception and a poor pregnancy prognosis . In our The treatment to be chosen for childbearing women with RA must not only be effective in controlling the condition but also safe and without any adverse effects on fertility and pregnancy. In the past few years, it was demonstrated that some of the anti-rheumatic drugs do not have a negative impact on pregnancy and if properly managed, the maternal and fetal risks are reduced to a minimum . In our In the past decade, biologic disease-modifying anti-rheumatic medication has notably improved the RA patient’s quality of life. Anti-TNFα agents are the most studied among the biological therapies not only for women but for men with autoimmune diseases, and they can be considered safe when used during the preconception period, as well as in the early stages of pregnancy . HoweverGlucocorticoids (GCs) were more frequently used during pregnancy than in the 3 months prior to conception, with most of the patients treated with GCs in the third trimester . There was a significant difference between the two subgroups regarding steroid intake during pregnancy, showing that this therapy improved the chance of having a pregnancy without adverse outcomes. Recent studies have demonstrated that oral GCs have no teratogenic effect when used for pregnant patients , but a cThis study is the first prospective study on RA pregnancies in Romania, and as there has not been much research on this subject at the national level up until now, we hope that our analysis will provide crucial information about how pregnancy affects RA and vice versa for women in our country, providing doctors with more confidence in managing the disease. Given that the study is multicentric, the primary limitation is the lack of homogeneity in the interdisciplinary follow-up of pregnancies. Another limitation is the small number of patients who enrolled. There are patients included with two or more pregnancies, which can be a bias, as the fear and stress regarding RA activity in relation to pregnancy evolution can be higher for the first pregnancy. Further investigation is required to provide a more thorough description of the illness and its consequences on pregnancy and postpartum disease evolution.In this study, adverse pregnancy outcomes are more frequent for RA pregnancy than the control group. Moreover, RA women have a higher age at conception, a shorter pregnancy length, and neonates with lower birth weight. The disease activity improved during pregnancy in 20% of cases and no teratogenicity was noticed. There was no correlation between disease activity or anti-rheumatic modifying disease drugs administration and pregnancy outcomes. The APO mothers had a shorter pregnancy length and newborns with lower birth weights compared with the No APO mothers. Unplanned pregnancy and age older than 35 years at conception were the major risk factors for APO in the studied population. Corticotherapy was a protective factor for uneventful pregnancy. Additional research with a larger cohort size is needed to investigate risk factors for adverse pregnancy outcomes in RA patients, as well as to establish the disease course post-partum and long-term outcome in children born to RA mothers.


Zanthoxylum bungeanum essential oil (EOZB) as an extract of Zanthoxylum bungeanum has a range of pharmacological effects such as antibacterial, anti-inflammatory, and antioxidant. However, there were no relevant studies on the regulation of gut microbes by EOZB in ruminants. In this study, the effects of different doses of EOZB on the structure and distribution of microorganisms in the small intestine of small-tailed Han sheep (STH) were investigated by 16s rRNA gene sequencing technique. We found that with the intervention of EOZB. The differential bacteria of duodenal at the phylum level were Firmicutes, Bacteroidetes, Tenericutes and Proteobacteria, and genus level differential bacteria were Prevotella 1, Ruminococcus 2 and Eubacterium coprostanoligenes group. The differential bacteria of jejunal at the phylum level were Firmicutes, Bacteroidetes, Tenericutes and Proteobacteria, and genus level differential bacteria were Prevotella 1, Rikenellaceae RC9 gut group, Christensenellaceae R-7 group, Ruminococcaceae UCG-014, Saccharofermentans, Ruminococcaceae NK4A214 group and Prevotellaceae UCG-001. The differential bacteria of ileal at the phylum level were Firmicutes, Bacteroidetes and Tenericutes, and genus level differential bacteria were Prevotella 1, Christensenellaceae R-7 group, Romboutsia and Ruminococcaceae UCG-014. In addition, at the same dose of EOZB, the five most abundant genera of bacteria varied in different regions of the small intestine. Among them, the abundance of Prevotella 1, Christensenellacea R-7 group and Ruminococcus 2 in ALW group was the highest in jejunum, duodenum and ileum, respectively. The abundance of Prevotella 1, Christensenellacea R-7 group and Rikenellacea RC9 gut group in BLW group was the highest in duodenum, jejunum and ileum, respectively. The abundance of Prevotella 1, Christensenellacea R-7 group and Ruminococcaeae NK4A214 group in CLW group was the highest in jejunum, duodenum and ileum, respectively. The abundance of Prevotella 1, Ruminococcus 2 and Ruminococcus NK4A214 groups in DLW group was the highest in jejunum, duodenum and ileum, respectively. Differential bacteria formed under the regulation of EOZB are associated with the digestion and absorption of nutrients and the state of intestinal health in the host. This study is the first to investigate the effect of EOZB on the distribution and structure of bacteria in the small intestine of STH. The results of the study enriched the structure and distribution of bacteria in the small intestine of ruminants and provided new insights into the future application of herbal medicine in ruminant production. Additionally, it provides a theoretical basis for the selection of probiotic bacteria for ruminants and the development and application of microecological preparations. Ruminants are essential for humans because they transform plant fibers into milk and meat . In contZanthoxylum bungeanum (Rutaceae) is currently widely distributed in most of China and some Southeast Asian countries. Zanthoxylum bungeanum is rich in flavonoids, primarily including rutin, quercetin, foeniculin, hyperin, and isoquercitrin  is a viscous liquid derived from plant husks with a distinct flavor that has pharmacological properties similar to those of Z. bungeanum  samples under the EOZB intervention by 16s rRNA high-throughput sequencing. Aimed to supplement the effects of EOZB on ruminant small intestinal bacteria and the incomplete of ruminant gut bacteria.p > 0.05) were obtained from a farm in Dingxi, Gansu Province, China. Test sheep were housed in individual pens that were regularly disinfected and cleaned (once a week). The 20 experimental sheep were randomly divided into four groups and provided the diets shown in Anatomy and Histology and Embryology of Domestic Animals ligation was performed at the division points of each part of the small intestinal tract, and the small intestinal tissue at each part was incised with a scalpel at the middle of the ligated segment. The contents of each part of the small intestine  were collected in 1.5-ml sterile polypropylene tubes, immediately frozen in liquid nitrogen at −80°C, and stored for analysis of bacterial communities.Twenty STH  according to the manufacturer’s protocol. The V4-V5 region of the bacterial 16S ribosomal RNA gene was amplified by PCR  using primers 515F (5′-GTGCCAGCMGCCGCGG-3′) and 907R (5′-CCGTCAATTCMTTTRAGTTT-3′), with the bar code a six-base sequence unique to each sample. The PCR reactions were performed in 30 μl of a mixture containing 15 μl of 2 × Phanta Master Mix, 1 μl of each primer (10 μM), and 20 ng of template DNA. Amplicons were extracted from 2% agarose gels and purified using an AxyPrep DNA Gel Extraction Kit  according to the manufacturer’s instructions. Purified PCR products were quantified by Qubit13.0 , and every 20 amplicons with different bar codes were mixed equally. The pooled DNA product was used to construct an Illumina paired-end library following the Illumina genomic DNA library preparation procedure. The amplicon library was then pair-end sequenced (2 × 250) on an Illumina Novaseq 6000 platform  according to the standard protocol.Raw data returned by the Illumina HiSeq sequencing platform were filtered using the software packages Pandaseq , PRINSEQp < 0.05 indicating significant differences.One-way ANOVA was performed using SPSS software . Analyzes were conducted on relative abundances and alpha diversity indices of bacteria in different parts of the small intestine at different levels. Multiple comparisons between groups were performed using Duncan’s method. Results are expressed as the mean and SEM, with A total of 1,992,846 tags were obtained from 20 duodenum samples; 1,873,850 clean tags were obtained after filtering, optimization, and quality control; and 78,484 sequences were extracted from each sample for subsequent analysis based on the minimum value of detected sequences . ClusterFirmicutes, Bacteroidetes, Tenericutes, and Proteobacteria, which had relative abundances >1% in all groups (Firmicutes was significantly higher in DLW than in the other groups (p < 0.001), whereas that of Bacteroidetes was significantly higher in ALW than in the other groups (p < 0.001). Relative abundance of Tenericutes was significantly higher in CLW than in the other groups (p = 0.034), whereas that of Proteobacteria was significantly higher in BLW than in the other groups ; whereas relative abundances of Ruminococcus 2 and Eubacterium coprostanoligenes group were significantly higher in DLW than in the other groups  based on Bray–Curtis distance showed significant differences in bacterial taxa among the groups . Anosim A total of 1,783,198 tags were obtained from 20 jejunum samples; 1,652,166 clean tags were obtained after filtering, optimization, and quality control; and 72,109 sequences were extracted from each sample for subsequent analysis based on the minimum value of detected sequences . ClusterFirmicutes, Bacteroidetes, Tenericutes, Proteobacteria, and Spirochaetes, which had relative abundances >1% (Firmicutes was significantly higher in DLW (p < 0.001); that of Bacteroidetes was significantly higher in CLW (p < 0.001); that of Tenericutes was significantly higher in BLW (p = 0.002); and that of Proteobacteria was significantly higher in ALW , those of Christensenellaceae R-7 group, Ruminococcaceae UCG-014, and Saccharofermentans were significantly higher in BLW , and those of Ruminococcaceae NK4A214 group and Prevotellaceae UCG-001 were significantly higher in ALW  and that of Bacteroidetes was significantly higher in ALW , that of Romboutsia was significantly higher in ALW (p < 0.001), and that of Ruminococcaceae UCG-014 was significantly higher in CLW  in different sites of the small intestine.Prevotella 1, Rikenellaceae RC9 gut group, Christensenellaceae R-7 group, Prevotellaceae UCG-001, and Saccharofermentans. In the jejunum, the five most abundant genera were Prevotella 1, Rikenellaceae RC9 gut group, Ruminococcaceae NK4A214 group, Prevotellaceae UCG-001, and Christensenellaceae R-7 group. In the ileum, the five most abundant genera were Prevotella 1, Romboutsia, Rikenellaceae RC9 gut group, Christensenellaceae R-7 group, and Saccharofermentans. In addition, the same genus of bacteria was distributed differently in different parts of the small intestine. The highest abundance of Prevotella 1 was in the jejunum, that of Christensenellaceae R-7 group was in the duodenum, and that of Ruminococcus 2 was in the ileum . Interestingly, Bacteroidetes decreased significantly in the DLW groups (p < 0.001). Firmicutes primarily include many gram-positive bacteria, including Lactococcus, Listeria, Bacillus, and Lactobacillus  ratio helps a host to absorb energy and maintain microbiota homeostasis . Prevotella 1 can degrade fiber-derived feeds such as hemicellulose or xylan . Spirochaetes were closely associated with fat metabolism pathways . Rikenellaceae RC9 gut group belongs to Rikenellaceae and most can ferment unabsorbed polysaccharides in the host gut to produce short-chain fatty acids such as acetate, propionate, and butyrate  production. Christensenellaceae R-7 group is in the Firmicutes . The functions of those phyla of bacteria are discussed in the section on the duodenum. At the genus level, Prevotella 1, Romboutsia, Christensenellaceae R-7 group, Rikenellaceae RC9 gut group, Turicibacter, Ruminococcaceae NK4A214 group, Saccharofermentans, Clostridium sensu stricto 1, Ruminococcus 2, Eubacterium coprostanoligenes group, Ruminococcaceae UCG-011, Ruminococcaceae UCG-014, Paeniclostridium, and Prevotellaceae UCG-001 were the dominant bacteria in the ileum of STH. Relative abundances of Prevotella 1, Christensenellaceae R-7 group, Romboutsia, and Ruminococcaceae UCG-014 were significantly higher in DLW, DLW, ALW, and CLW, respectively, than in the other groups . Romboutsia is significantly positively correlated with total carbohydrate digestibility  of the five most abundant genera of bacteria was different in different parts of the small intestine  at the same dose of EOZB. Some genera were specific to different sites in the small intestine at the same dose of EOZB. The differential genera of bacteria are associated with the digestion and absorption of host proteins, carbohydrates, and lipids. Simultaneously, those bacteria can have positive effects on the development of host intestinal epithelial cells, maintenance of microbial and metabolic homeostasis, and control of inflammatory processes. Thus, although the evidence is indirect, it indicates that EOZB affects the above physiological processes. Our findings fill the gap of bacteria composition structure of small intestine in ruminants, and also provide some theoretical basis for the selection of probiotic bacteria and the development and application of EOZB.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/The animal study was reviewed and approved by Experimental Animal Ethics Committee of Gansu Agricultural University . Written informed consent was obtained from the owners for the participation of their animals in this study.CW and XL conceived, designed, and supervised the project. HZ, XL, and YZ collected samples and performed experiments. HZ and XL carried out bioinformatic analyzes. HZ drafted the manuscript. HZ, XL, YZ, and CW revised the manuscript. XL, HZ, and CW, contributed ideas on the paper. All authors have read, edited, and approved the final manuscript.This work was supported by the Project of Research Condition Construction and Achievement Transformation of Gansu Academy of Agricultural Sciences .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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.