Diabetes mellitus (DM) is one of the most common diseases worldwide. DM may disrupt hormone regulation. Metabolic hormones, leptin, ghrelin, glucagon, and glucagon-like peptide 1, are produced by the salivary glands and taste cells. These salivary hormones are expressed at different levels in diabetic patients compared to control group and may cause differences in the perception of sweetness. This study is aimed at assessing the concentrations of salivary hormones leptin, ghrelin, glucagon, and GLP-1 and their correlations with sweet taste perception (including thresholds and preferences) in patients with DM. A total of 155 participants were divided into three groups: controlled DM, uncontrolled DM, and control groups. Saliva samples were collected to determine salivary hormone concentrations by ELISA kits. Varying sucrose concentrations  were used to assess sweetness thresholds and preferences. Results showed a significant increase in salivary leptin concentrations in the controlled DM and uncontrolled DM compared to the control group. In contrast, salivary ghrelin and GLP-1 concentrations were significantly lower in the uncontrolled DM group than in the control group. In general, HbA1c was positively correlated with salivary leptin concentrations and negatively correlated with salivary ghrelin concentrations. Additionally, in both the controlled and uncontrolled DM groups, salivary leptin was negatively correlated with the perception of sweetness. Salivary glucagon concentrations were negatively correlated with sweet taste preferences in both controlled and uncontrolled DM. In conclusion, the salivary hormones leptin, ghrelin, and GLP-1 are produced either higher or lower in patients with diabetes compared to the control group. In addition, salivary leptin and glucagon are inversely associated with sweet taste preference in diabetic patients. Diabetes mellitus (DM) is one of the most common diseases all over the world. DM is caused by an insulin deficiency or resistance and characterised by hyperglycaemia. The latest statistics reported that in 2021, more than 537 million individuals around the world have diabetes, and this number is expected to increase to 783 million by the year 2045 [α-cells of the islets of Langerhans. Its main function is to increase blood glucose concentrations. It is also expressed in a subset of taste cells of the tongue [DM has severe complications such as atherosclerotic, heart failure, chronic kidney, retinopathy, and neuropathy diseases . Moreovee tongue . Finallye tongue . In addie tongue . These se tongue .Taste perception, in general, is essential for living; it conveys information about food and influences food selection and dietary intake , 13. The∗Power software, version 3.1.9.7) that indicated a 95% chance of a 0.5 effect size between the tested groups at the 5% level (two-tailed). The participants were divided into three groups according to their glycated haemoglobin (HbA1c) concentrations. The participants in the control group (n = 52) had HbA1c concentrations of less than 5.7%, those in the controlled DM group (n = 52) had HbA1c concentrations of less than 7%, and those with uncontrolled DM (n = 51) had HbA1c concentrations of ≥7%. HbA1c data was obtained from participants' medical files, for diabetic within 3 months of test day and for control group within 6 months of test day. Patients who had hypothyroidism, stroke, and cancer or were pregnant and smokers, used antibiotics, had oral lesions, had influenza or COVID-19, had been exposed to radiation in the head or neck regions, or were under 35 years or above 65 years of age were excluded . In addition, each participant's waist and hip circumferences were measured using a fabric measuring tape, and these measurements were used to calculate each participant's waist-to-hip ratio (WHR).g for 20 minutes. The supernatant was then collected and stored at -80°C until the analysis. The salivary hormone concentrations were determined using sandwich enzyme-linked immunosorbent assay kits , according to the manufacturer's protocol. Human Leptin ELISA kit (E1559Hu), Human Ghrelin ELISA kit (E1552Hu), Human Glucagon ELISA kit (E1023Hu), and Human Glucagon-like Peptide-1 ELISA kit (E0022Hu) were used to determine leptin, total ghrelin, glucagon, and GLP-1 concentrations, respectively.A saliva sample was collected from each participant in an odourless room during morning (from 8 AM to 11 AM). Each participant fasted for one hour prior to the sample collection. The participants rinsed their mouths with distilled water to remove any excess food. Next, the head was tilted forward for some minutes, and a 2 ml of saliva was collected (without stimulators) in a sterilised tube from the anterior of the mouth and placed directly on ice. Then, the saliva samples were centrifuged at 1000 × Each participant's sweet taste perception including sweet taste threshold and sweet taste preference was assessed. The sweetness threshold  was determined in the following manner: the participant was offered different concentrations of a sucrose solution , in ascending order, until they identified a sweet taste . After eP value of < 0.05 was considered statistically significant.One-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test was used to assess differences in the concentrations of leptin, ghrelin, glucagon, and GLP-1 between the three groups . In addition, this test was also used to determine the differences in the sweetness thresholds and preferences between the three groups. The relationships between each salivary hormone and HbA1c, sweet taste threshold, and sweet taste preference were tested using the Spearman rank correlations. All statistical analyses were performed using GraphPad Prism software . A 2). The mean WHR was 0.86 ± 0.22, and the mean HbA1c was 6.73 ± 1.70%. The participants in the controlled and uncontrolled DM groups had diabetes for a mean duration of 10.04 ± 8.64 and 13.23 ± 7.41 (years), respectively. In the control group, 54% of the participants has been diagnosed with hypertension. This percent was slightly higher in controlled DM and uncontrolled DM . With regard to sweet intake, 40%, 53%, and 63% of the participants in the control group, controlled DM, and uncontrolled DM did not prefer sweet food over other foods, respectively. Although almost half of the participants in each group always crave sweets, more than half of them did not consume much sweet food. Moreover, 82% of the participants in the control group believed that sweet foods affect their health. Similar belief was also found in diabetic patients.This study included a total of 155 participants; their demographic characteristics are shown in The mean concentrations of leptin, ghrelin, glucagon, and GLP-1 in each of the three groups are shown in The differences in the sweet taste thresholds and preferences between the three groups  are shown in The correlation between salivary leptin, ghrelin, glucagon, and GLP-1 concentrations and HbA1c, sweet taste thresholds, and sweet taste preferences for all three groups are shown in The purpose of this study was to assess the concentrations of the salivary hormones leptin, ghrelin, glucagon, and GLP-1 and their relationships with the perceptions of sweetness in individuals with controlled and uncontrolled DM as compared with control individuals.The results of our study demonstrated that salivary leptin concentrations were higher in the uncontrolled and controlled DM groups than in the control group. In addition, there was a strong positive correlation between salivary leptin and HbA1c in all groups. This finding is consistent with the finding of a previous study which reported that salivary leptin was twofold higher in DM patients compared to healthy controls. Moreover, they found a positive significant relationship between salivary leptin and insulin and some inflammatory markers. These correlations may indicate that the increment in salivary leptin observed in our study could be due to the situation of insulin resistance and inflammation occurring in DM, since leptin can act as proinflammatory cytokine . In contIn our study, the three groups showed no differences with respect to sweet taste thresholds and preferences. Similarly, previous study conducted by Wasalathanthri et al. reported no difference in sweet recognition threshold between patients with normal glycaemic levels and those with diabetes . FurtherATP channels that are expressed in taste cells which in turn suppress their responses to sweet compounds [Our results indicated that in the controlled DM group, salivary leptin was negatively correlated with sweet taste threshold and sweet taste preference; similar findings were also seen for the uncontrolled DM group with respect to sweet taste preference. These results are similar with the previous results conducted on mice which showed a strong correlation between leptin and sweet taste perception. Leptin suppresses sweet taste via leptin receptor (Ob-Rb) and Kompounds . As for ompounds . In addiompounds , while sompounds .To the best of our knowledge, this study is the first to report the associations between salivary leptin, ghrelin, glucagon, and GLP-1 concentrations and sweet taste perceptions, including sweet taste thresholds and preferences, in individuals with controlled DM, those with uncontrolled DM, and those without DM. The salivary hormones leptin, ghrelin, and GLP-1 are either higher or lower in patients with diabetes compared to the control group. In addition, salivary leptin and glucagon are inversely associated with sweet taste preference in diabetic patients. However, a number of limitations need to be considered, for instance, measurement of the expression of taste receptors in the tongue. Thus, future research should be done to investigate the mechanism of action of these salivary hormones on sweet taste and the expression of taste receptors.


DNMT) as well as histone deacetylase (HDAC) families. Following the exposure, organ biopsies were collected from the lungs, kidneys, liver, and spleen, and the gene expression levels were determined by real-time quantitative reverse transcription PCR (RT-qPCR). Our results demonstrated significant upregulation of genes in the tested organs in various patterns followed by the intake of tartrazine on ADI. Since DNMT and HDAC genes are involved in different steps of carcinogenesis, have roles in the development of neurological disorders and the effect of dose of everyday exposure is rarely studied, further investigation is warranted to study these possible associations.In recent years, artificial additives, especially synthetic food colorants, were found to demonstrate wider properties compared to their natural equivalents; however, their health impact is still not totally mapped. Our study aimed to determine the long-term (30 and 90 days) exposure effect of one of the commonly used artificial food colorants, tartrazine, on NMRI mice. The applied dose of tartrazine referred to the human equivalent dose for acceptable daily intake (ADI). Further, we evaluated its impact on the transcription of a range of epigenetic effectors, members of the DNA methyltransferase ( Color additives have been widely used since 1500 BC and currently are extensively used by food manufacturers as a vital criterion for food choice. Among them, synthetic food colorants are more favorable due to their stability, low cost, and coloring properties. These artificial food colorants are available in different varieties and colors .The consumption of artificial food colorants below the acceptable daily intake (ADI) does not cause any harm, but it may cause for example many disorders among children due to their low body weight . In deveAzo dyes are a group of vivid synthetic food colorants including tartrazine -4-(4-sulfonatophenyl-azo)-H-pyrazol-3-carboxylate) and represent about two-thirds of all synthetic dyes, which are undoubtedly the most widely used colorants in our everyday products . Red azoTRZ is synthesized from coal tar, and it is marketed in the form of water-soluble powder. The levels of TRZ and other synthetic colorants in food product samples can exceed the accepted daily consumption limit in adults and children. Among children, the risk of exceeding ADI is usually much higher than adults, because children are the major consumers of colored food. Thus, exposure to excessive colorants may pose a greater health risk such as hyperactivity . The indTRZ can be metabolized to an aromatic amine such as sulfanilic acid, which is highly sensitizing to allergies such as urticaria and asthma because TRZ is a nitrous derivative ,14. In ap < 0.05), as well as inhibition of glutathione (GSH) concentration, catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) antioxidant enzyme activities. Additionally, tartrazine-treated rats exhibited elevated levels of acetylcholine (Ach) and gamma-aminobutyric acid (GABA) in the brain, while dopamine (DA) levels were depleted [TRZ has been demonstrated to exert histopathological effects on the hepatic and renal tissues of rats, indicated by vacuolation, swelling, necrosis, and pyknosis . Red-azodepleted .DNA methyltransferases (DNMTs) and histone deacetylases (HDACs) are groups of enzymes that play a significant role in epigenetics. DNMTs are a family of enzymes that provide a crucial role in genomic stability and integrity. They act on the addition of a methyl group to the 5-position of cytosine residues in DNA. DNA methylation plays a crucial role in regulating gene expression by silencing certain genes by inhibiting their transcription. This can occur through the methylation of promoter regions of genes, preventing the binding of transcriptional factors . There aHDACs represent a group of enzymes responsible for the removal of acetyl groups from histones, leading to expressions of several genes. It has been reported that HDACs are responsible for the alteration of acetylation levels, which has been studied in various cancer cells. HDACs could play a significant role in the initiation, progression, and promotion of carcinogenesis .HDAC genes inhibit cell cycle inhibitors, differentiation, and apoptosis but enhance angiogenesis, invasion, and migration [HDAC2, HDAC3, and HDAC8 are all members of the histone deacetylase (HDAC) family of enzymes that play important roles in regulating cell proliferation. They are known to interact with other transcriptional repressors and co-repressors, such as the polycomb group proteins, to silence genes that promote cell proliferation. Studies have shown that igration ,32.Many food products contain TRZ, which are prepared at different temperatures, such as drinks and juices, cookies, chips, chewing gums, candies, cereals, mustards, dairy products, jellies, ice creams, fillings, liqueurs, powdered juices, soft drinks, yogurts, decoration and coatings, and many other products .HDAC2, HDAC3, and HDAC8 as well as DNMT1, DNMT3a, and DNMT3b with the help of qRT-PCR measured in the lungs, kidneys, liver, and spleen of young male and female mice after long-term and short-term exposure, resulting in a detectable enhancing effect of tartrazine on the activity of DNMT and HDAC genes.This study aimed to evaluate the correlation between TRZ and the expression of The selected age of the mice was 6–8 weeks. This corresponds to the human infancy and early sedentary age. The duration of treatment was 30 and 90 days for the mice, which corresponds to approximately 3 and 10 years in human life. Thus, our study targets the life stage and duration at which exposure to tartrazine is greatest in the human population. A total number of 48 male and 48 female NMRI mice  were used in the present study, aged between 6 and 8 weeks, and weighing 30–40 g. The study animals were kept at the Experimental Department , where they were housed in standard polycarbonate cages (330 × 160 × 137 mm), bedded with shavings under standard conditions  and fed with standard rodent pellet ; the water was provided ad libitum. The animal experiment was reviewed and approved by the local authorities  according to Hungarian animal protection laws in accordance with EU guidelines. In line with ethical guidelines, we reduced the number of animals used and refined the experimental conditions and procedures to minimize harm to animals. The cervical dislocation was performed by a properly trained person in full compliance with the regulations of physical euthanasia.NMRI mice were divided into 8 groups of 6 females and 6 males each as described below:Group I: control group that consumed tap water and rodent chew prepared at room temperature for 30 days.Group II: control group that consumed tap water and rodent chew prepared at room temperature for 90 days.Group III: control group that consumed tap water and rodent chew, prepared, and baked after the drying process for 30 min at 160 °C, for the duration of 30 days.Group IV: control group that consumed tap water and rodent chew, prepared, and baked after the drying process for 30 min at 160 °C, for the duration of 90 days.Groups V and VI: received special feed consisting of 1×ADI equivalent of human dosage   of TRZ  according to the manufacturer’s protocol. The tissue samples were homogenized using a Polytron homogenizer with TRIzol reagent (1 mL per 50–100 mg of tissue), and 100 µL of chloroform was added to the TRIzol lysate and thoroughly mixed by shaking. After 5 min at room temperature, the samples were centrifuged for 15 min at 12,000× HPRT1), and the genes of interest, DNMT1, DNMT3A, DNMT3B, HDAC2, HDAC3, and HDAC8, are shown in −∆∆CT) method.Total RNA was evaluated with Roche Lightcycler 480 . Assays were run with the Roche 480 instrument, using the KAPA SYBR FAST One-Step qRT-PCR Master Mix kit . Amplifications were carried out in 20 μL of reaction volume, mixing 5 μL of RNA target (50–100 ng) and 15 μL of the master mix containing forward and reverse primers , 3.8 µL sterile double-distilled water). Reactions were performed with the following thermal profile: 42 °C for 5 min, for the reverse transcription step, a hot-start denaturing step of 95 °C for 3 min, followed by 45 cycles of 95 °C for 10 s (denaturation), 60 °C for 20 s . The fluorogenic signal emitted was read during the annealing–extension step and was analyzed by software. Immediately after amplification, a melting curve protocol was produced by increasing each cycle by 0.5 °C, starting from the set-point temperature (55.0 °C), for 80 cycles, each one at 10 s. The primary sequences of the housekeeping gene used as an internal control, hypoxanthine–guanine phosphoribosyltransferase . The Kolmogorov–Smirnov test was applied to determine the distribution and standard deviation. Statistical analysis was performed using IBM SPSS statistics for Windows, Version 26.0 . Significance was established at DNMT1 expression . Additionally, there was a significant reduction in the total antioxidant capacity observed following tartrazine treatment [DNMTs, which can result in DNA hypermethylation [DNMT1, DNMT3a, HDAC2, and HDAC3 in both liver and kidneys along with HDAC8 in the kidney tissues, but the level of DNMT3b did not change noticeably in the kidney.To study the long-term effects, Himri and colleagues (2011) found that ADI doses of TRZ and its consequent metabolic byproducts, namely, sulfanilic acid, have the ability to produce reactive oxygen species (ROS), which leads to oxidative stress and potentially impacts the hepatic and renal structures as well as their respective biochemical profiles . Adminisreatment . The increatment . Relatedhylation . In the DNMT activity in the epithelial cells [DNMTs and HDACs, which suggests that tartrazine may have a negative effect on lung health by inducing oxidative stress, inflammation, and upregulation of pro-inflammatory cytokines. However, further research is needed to fully understand the mechanisms underlying these effects.There is limited research on the specific molecular pathways in the lungs that may be affected by tartrazine. However, some studies have suggested that tartrazine may induce oxidative stress and inflammation ; howeveral cells . In currHDACs and DMNTs overexpression in the brain. One study conducted by Valera et al. suggested that modulating the DNMT and HDAC enzymes could potentially contribute to the manifestation of anti-manic effects. This was concluded because effectively regulating these enzymes has been observed to reverse the effects of Ouabain (OUA), an inhibitor of Na+/K+ATPase. OUA is known to extend the depolarized state of neurons and to induce hyperactivity [GSK-3 and HDACs [HDAC and DNMT genes can alleviate conditions such as mania, hyperactivity, bipolar disorders, neurological disorders, and behavior deficits, which can be reversed with medications that act as HDAC inhibitors, such as valproate. In the current study, we witnessed an overexpression of these genes in several organs due to the exposure of the experimental animals to tartrazine; meanwhile, tartrazine and its metabolites can pass the blood–brain barrier and induce neurotoxicity. Although we gained data from different organs other than the brain, we hypothesize that tartrazine may play a role in behavior and neurological disorders from consuming a food product that meets the current regulations regarding ADI. We believe that further investigations of everyday doses of tartrazine are needed to study the long-term impact on brain function.Recently, there have been several studies published regarding the effect of epigenetic mechanisms such as activity . Anothernd HDACs . Wang etnd HDACs . The menDNMT and HDAC genes over a long treatment period while keeping the recommended dose of intake. The present study shows that tartrazine can cause a change in the gene expressions of DNMT1, DNMT3a, DNMT3b, HDAC2, HDAC3, and HDAC8 over a long period even at a dose at the ADI level in different organs. Based on the data, compared with the literature, we hypothesize that tartrazine consumption may contribute to cell proliferation and may even increase the likelihood of carcinogenesis. Furthermore, the observed effect in various organs highlights the possibility that the genes under investigation may also be activated in the brain, with proven consequences for brain function. This is the first study to investigate the effects of oral administration of tartrazine on In view of these findings, it is of great importance to investigate the direct effect of tartrazine on brain function, and replacing the artificial mono-azo dyes with natural food dyes in the food industry, where it is necessary to color food and beverages, should be considered.


Alleviation of national-scale planetary boundary transgression has been achieved primarily in the US, China, Saudi Arabia, etc., while aggravation was incurred in Pakistan, Australia, Argentina, and so forth.Food systems are among the leading causes for transgression of planetary boundaries globally, which define the safe operating space for humanity. We quantify unsustainable environmental impacts of food systems, indicated by the transgression of national-scale planetary boundaries , from both production and consumption perspectives of 189 countries/regions around the world. A multi-regional input-output model is used to map the global transfers of the national-scale transgression of planetary boundaries, including freshwater use, land change, and biogeochemical flows (nitrogen and phosphorus). Our results show that China is a major global unsustainable water and nitrogen exporter and an unstable land and phosphorus importer. This means that water and nitrogen uses in China are used to support food demands in other countries, and food consumption in China requires unsustainable land and phosphorus uses elsewhere. In contrast, the US is a major exporter of unsustainable water, land, and nitrogen uses but only an importer of unsustainable phosphorus for food consumption. Globally, compared to a counterfactual scenario where there is no food trade among any countries, food trade saves massive transgressions of planetary boundaries (270 km  •Unsustainable water and nitrogen use in China and the US support food demands elsewhere•Globally, food trade avoids substantial national planetary boundary transgressions•Trade alleviates national planetary boundary transgressions in the US and China•Trade aggravates national planetary boundary transgressions in Pakistan and Australia Agricultural land; Environmental science; Food policy; Sustainability aspects of food production Rockstrom et al. (2009)2 of cropland use, 69 teragrams (Tg) of nitrogen application, and 16 Tg of phosphorus application.Production of food to meet humankind’s rising food demand is a major contributor to the transgression of those local boundaries since agricultural activities in many regions are using too much water, land, or fertilizer.,Meanwhile global trade of agricultural and food products may change the pattern of those local problems. For example, by importing water-intensive wheat-based and corn-based products from water-abundant countries, such as Brazil, water-scarce countries in the Middle East seek to alleviate their local water stress.However, few studies have examined whether those embodied environmental impacts are sustainably within the planet’s carrying capacity especially at the place of production, indicated by their local or national planetary boundaries.This study thus makes novel contribution by exploring the global transfers of transgression of planetary boundaries downscaled to the national level, i.e., national boundaries, to meet growing human food demand combining the downscaling of global planetary boundaries and quantification of consumption-based environmental footprints. How the global transfers of national boundary transgressions have helped alleviate or aggravated those national boundary transgressions have also been examined. Although Vanham et al. (2019)3 in 1995 to 713 km3 in 2015, with top 10 countries occupying almost 90 percent. India has used 276 km3 of water beyond its national boundary for food production in 2020, primarily for cultivation of wheat and paddy rice, accounting for almost 39 percent of the global total, followed by China (111 km3), Pakistan (74 km3), the US (50 km3), and Egypt (45 km3), which remained the top 5 countries with the largest amounts of unsustainable water use for food production in both 1995 and 2015  in 1995 to almost 50 percent (15 million tons) in 2015, primarily to grow vegetables, paddy rice, grains, and wheat. China (6 million tons), India (4.6 million tons), and Brazil (4.5 million tons) were the three largest unsustainable phosphorus users for food production, together making up 55 percent of the global total. Besides vegetable and paddy rice cultivation in India and China, cattle farming is also a main contributor of unsustainable phosphorus application in Brazil.2 to 23 million km2.The US, China, Brazil, Australia, and Kazakhstan were top five countries where food production has occupied the largest amounts of unsustainable land use in both 1995 and 2015, together accounting for more than 30 percent of the global total, primarily used for cattle farming. Another two issues worth noting are the following: first, top ten countries contributed about 57 percent of the global total; secondly, unlike the other three planetary boundary indicators, global total of national unsustainable land use for food production has decreased from 28 million km3, respectively, around the world to meet their respective food demands, followed by the US (49 km3) and Pakistan (42 km3). Food demand of the Chinese population is produced with the largest unsustainable nitrogen footprints globally (14 million tons), which made up 44 percent of the global total, followed by India and the US.Increases in food production and its unsustainable environmental footprints, defined as environmental impacts along the life cycle supply chain, are incurred to support food consumption both domestically and from other countries. We quantify the extent to which food demands in one country are produced from supply chain that require excessive and unsustainable use of national resources (land and water) and nutrition flows (nitrogen and phosphorus) either domestically or elsewhere in other countries. As shown in 2 of land transgressing national boundaries around the world, largest among all countries studied.Different from water and nitrogen, where China, India, and the US led the rank, Brazil accounted for more food-driven unsustainable phosphorus footprints than the US. Similarly, food demands in China, the US, and Brazil required, respectively, 4, 3, and 1 million km3 of unsustainable water footprints transgressing national boundaries around the world, equaling to the world average, while equaling to 40 percent of that of Indians (196 m3) and half of that of Americans (153 m3). In contrast, China has the highest per capita unsustainable nitrogen footprints (10 g per capita), more than twice of the global average (4 g per person). From 1995 to 2015, consumption-based unsustainable footprints per capita have increased for all indicators around the world, with country rankings remaining stable.However, as food security is considered as a basic human right, consumption-based national boundary transgression should also be viewed in per capita terms. For example, every Chinese person’s food consumption required 80 m3), the US to Georgia (4 km3), and Pakistan to Georgia (3.5 km3).As shown in It is noteworthy that compared to 1995, the major unsustainable water use exporters remained almost the same, including China, Egypt, India, Pakistan, Thailand, and the US, who are major water-intensive food exporters, exporting rice, soy, fruits, and so forth. However, the composition of importers has substantially diversified. In 1995, the major importers were almost all large and developed economies, including the US, Japan, Saudi Arabia, Germany, Italy, the UK, Russia, etc, while in 2015, a number of emerging and less developed economies have emerged as primary contributors to transgression of NBW elsewhere.2), the US (292 km2), Brazil (188 km2), and Australia (163 km2). In total, China accounted for 8 out of the top 20 flows, as well as about 57 percent of the transferred amounts.Australia, Mongolia, the US, Argentina, and Canada have remained the five major exporters of unsustainable land use for food productions in both 1995 and 2015. The major importers also remained almost the same, including China, the US, and Japan. Japan and the US were the primary importers in 1995, while China solely dominated the imports in 2015. In 2015, top four unsustainable land flows were all exported to China, from Mongolia (341 kmAlthough China dominated the production-based unsustainable nitrogen uses (section 3.1), large amounts of those were used to produce food to support consumption in other countries, for instance, Japan, South Korea, and Germany. Similarly, transgression of the National Boundary for Nitrogen (NBN) in the US is also incurred to support food consumption in Japan, South Korea, and Mexico. In contrast, food demands in China and the US are produced from production activities that require unsustainable phosphorus applications in Brazil, India, Argentina, and so on.3, NBN by 13 million tons, and National Boundary for Phosphorus (NBP) by 5 million tons. Next to the US, China also benefitted from the global food trade system reducing its NBW transgression by 43 km3, NBN transgression by 2 million tons, NBP by 1 million tons, NBL by 1,839 million km2. Saudi Arabia reduced 2,287 million km2 of NBL transgression, the highest among all countries through global food trade. The UAE had the largest NBW transgression alleviation, amounting to 88 km3 in 2015.We compare the transgression of national boundaries for food systems with a counterfactual scenario where no global food trade system exists and all countries are self-sufficient in food. As shown in 3 in Pakistan is used to produce food products that are exported to other countries. Such model findings are in line with several previous studies. For instance, Dalin et al. (2017)3 from 1990 to 2015. Global food trade has resulted in 949 million km2 of additional NBL transgression in Australia, the largest among all countries, followed by 713 million km2 in Argentina.On the other hand, national boundary transgressions in some countries, including Pakistan, are to support the global food trade system. An NBW transgression of 28 km3 of unsustainable water use worldwide, 18 million tons of unsustainable nitrogen use, 7 million tons of unsustainable phosphorus use, and 5,431 million km2 of unsustainable land use for food compared to a counterfactual scenario where all foods consumed are produced domestically Some predominant agricultural water-saving technologies include drip and intermittent irrigation, chemical-based sprays, and so forth.The environmental impacts of food systems could be minimized from both producer and consumer’s sustainable practices. From the consumer’s perspective, environmentally responsible food choices should be promoted. For instance, the increasing consumption of avocado by bourgeoning urbanites is imposing increasing water pressure along the life cycle supply chain.3 of water lost, and 1.4 billion hectares of land wasted.On other note, food demand management is of critical importance of reaping the double benefits of eliminating hunger while minimizing environmental disturbances.3) than in 1995 (218 km3), but less unsustainable nitrogen (18 million tons in 2015 and 29 million tons in 1995), phosphorus (7 million tons in 2015 and 13 million tons in 1995), and land uses (5.4 billion km2 in 2015 and 14.3 billion km2 in 1995). Literature review in the virtual water field has also concluded that food trade has generally led to global water savings.We quantify unsustainable environmental impacts of national food systems from both production and consumption perspectives of 189 countries/regions around the world with a global multi-regional input-output (MRIO) analysis. Unsustainable environmental impacts are quantified based on the exceedance of national planetary boundaries, and the cross-national transferred environmental impacts from consumers to producers are also evaluated. Unsustainable amounts of nitrogen were used in China and the US to support food consumption in many other countries, including Japan, South Korea, and others, in line with findings from Shi et al. (2016).Trade patterns can be optimized to internalize its environmental externalities. Although optimized trade needs to consider a large variety of factors, including predominantly capital, labor, and technology among others, environmental impacts, especially those that have already transgressed national boundaries where products are produced, should also be incorporated. National boundary transgression provides another important parameter measuring unsustainable environmental impacts induced by trade. Our results have shown that although environmental impacts of food productions are still well within national boundaries for some, i.e., Canada and North European countries, national boundaries of their trading partners, e.g., Brazil, have been seriously transgressed . On anot,However, it should be noted that there are several limitations and uncertainties in the methodology and therefore results achieved in this study, which could be further studied in future academic endeavors. First, MRIO uses sector average environmental impacts intensity data, which may generate uncertainties in estimating country-specific links and transfers. Secondly, downscaling from global to national values can be conducted through different ways, which all entail methodological uncertainties. Due to data availability, we distribute planetary boundary to national level according to farmland area. However, such approach can create perverse incentives for countries who have deforested a lot of land today than countries that retained their intact forests because the former gets higher shares of the planetary boundary. Some of these issues could be avoided in future studies, for example, by using regional planetary boundaries.chaili@cau.edu.cn).Further information and request for resources should be directed to and would be fulfilled by the Lead Contact, Li Chai , National Boundary for Land (NBL), National Boundary for Nitrogen (NBN), and National Boundary for Phosphorous (NBP).Similar with Hu et al. (2020),It should be noted that while some previous studies, such as Hu et al. (2020),A Multi-Regional Input-Output (MRIO) model is used to assess the global transfers of unsustainable environmental impacts  as virtual flows embodied in global food trade. Unsustainable environmental impacts are defined as those exceeded, or transgressed, national boundaries . We adopt a highly disaggregated MRIO database (189 countries/regions and 163 sectors) complied by Cabernard and Pfister (2020)The unsustainable environmental impacts, i.e. environmental footprints that have transgressed national boundaries (The global virtual flows of unsustainable environmental impacts can be assessed by the below In the scenario of “no trade”, we assume that all consumed foods were produced domestically, which can be calculated by


The latest outbreaks of high pathogenicity avian influenza viruses (HPAIVs) were the most widespread ever seen in Europe, entailing considerable economic costs and raising public health concerns. Virological surveillance protocols involve reverse transcription quantitative polymerase chain reactions (RT-qPCR) from tracheal and cloacal swabs, which are laborious to perform on a large scale and require special skills. Environmental sampling, and especially dust collection, may represent a relevant alternative, as it is cheap, non-invasive for animals, simpler, and quicker to carry out. The main drawback of dust samples is that they may contain high amounts of organic substances that can inhibit RT-qPCR reactions. Bovine serum albumin (BSA) is a molecule known to facilitate DNA polymerization in the presence of numerous inhibitors, including those from feces, litter, or food. We tested its use on dust samples collected on 107 farms localized in areas affected by epizootics of clade 2.3.4.4b HPAIV H5N1. We used a latent class modeling approach to compare the performance of three detection protocols: (i) BSA addition to the RT-qPCR reaction mix, (ii) dilution of template RNA, and (iii) a control protocol. Our results indicate that BSA addition to the RT-qPCR reaction mix improved the sensitivity of the method, by neutralizing inhibitors’ effect. Indeed, for hemagglutinin (HA) or matrix (M) RNA detection, the sensitivity of the BSA protocol was the highest, followed by that of the control protocol, and that of the dilution protocol. Our results suggest that the use of BSA could be routinely implemented in HPAIV dust monitoring RT-qPCR protocols.With the circulation of high pathogenicity avian influenza viruses having intensified considerably in recent years, the European Union is considering the vaccination of farmed birds. A prerequisite for this vaccination is the implementation of drastic surveillance protocols. Environmental sampling is a relevant alternative to animal sampling. However, environmental samples often contain inhibitory compounds in large enough quantities to inhibit RT-qPCR reactions. As bovine serum albumin is a molecule used in many fields to overcome this inhibitory effect, we tested its use on dust samples from poultry farms in areas heavily affected by HPAIV epizootics. Our results show that its use significantly increases the sensitivity of the method. High pathogenicity avian influenza viruses (HPAIVs) have become a major threat to the poultry industry and wild bird populations , 2. ThesThe number of epizootics caused by HPAIV is increasing worldwide . Their sThe main drawback of dust samples is that they often contain inhibitory substances in sufficient amounts to disrupt reverse transcription quantitative polymerase chain reactions (RT-qPCR) or cause fluorescence inhibition , 11. PCRWe took dust samples from 107 poultry houses at very high risk of HPAI to determine how the addition of BSA to the RT-qPCR mix would affect sensibility and sensitivity. Three protocols were compared: a control protocol, BSA addition, and 1:10 RNA dilution. Importantly, in this method-oriented study, we focused on the dust samples only, regardless of the other types of samples, including swabs.We conducted a study between December 2021 and April 2022, during the H5N1 high pathogenicity avian influenza epizootic, on a total of 107 French poultry houses. The selected farms were either confirmed HPAIV outbreaks or had a direct epidemiological link with other HPAIV-positive farms. Dust samples were collected using dry wipes rubbed against the walls of the buildings or the feed troughs.n = 21) and 16.8% (n = 18) were, respectively, HA and M positive with the control protocol , 26.2% (n = 28) and 25.2% (n = 27) were, respectively, HA and M positive with the BSA protocol, and 15.9% (n = 17) and 14.0% (n = 15) were, respectively, HA and M positive with the dilution protocol. All HA and M positive samples in the control protocol were also positive with the addition of BSA and in the dilution protocols. All HA and M positive samples from the dilution protocol were also positive with the BSA protocol. More importantly, 5.6% (n = 6) and 6.5% (n = 7) samples were, respectively, HA and M positive only under the BSA protocol , the Food and Agriculture Organization (FAO), and the French Agency for Food, Environmental, and Occupational Health and Safety (ANSES) for the detection of Eurasian H5 HPAIVs , 18. We protocol . DetaileBSA addition was not detrimental to the RT-qPCR reaction. On the contrary, we observed a slight but not statistically significant increase in viral RNA copy numbers compared to the control protocol. As expected, 1:10 dilution of RNAs resulted in a statistically significant decrease in viral RNA copy numbers compared with control conditions and the addition of BSA, for both the HA and M genes .4 50% tissue culture infectious dose (TCID50) to 107 TCID50, before performing RNA extraction and RT-qPCR  included conditional dependence between the control and dilution protocols in positive samples (DIC = 20.2). As no other models were within two points of difference in DIC from the best-fit model, this model was selected for inferring the parameter estimates of the proportion of infected farms and the sensitivity of each protocol.As illustrated in The relevance of environmental sampling for viral surveillance is increasingly considered and has been emphasized during the COVID-19 pandemic with wastewater-based surveillance, which provided very valuable information, in complement to individual-based PCR testing. It also raised the issue of sewage samples’ RT-qPCR inhibition and the need for optimization and controls in dedicated protocols , 21. In For matrices with high levels of amplification inhibitors, dilution of samples or extracted nucleic acids is a widely applied method, since it automatically results in an amplification inhibitors dilution, a 10-fold dilution being commonly used , 24. UnfThe principal limitation of our study is that we did not take cloacal and/or tracheal swabs on poultry farms, thus preventing us from directly comparing dust sampling with swabs. However, this comparison was carried out in a previous study, on a smaller number of poultry houses, showing that RT-qPCR from dust was an equivalent alternative regarding sensitivity .Dust sampling protocols for HPAIVs monitoring are set to become increasingly important in France and Europe, with the European Union imposing stringent monitoring protocols as a prerequisite for the roll-out of vaccination of poultry against H5Nx viruses . Such prAmong these potential additives, BSA is an inexpensive, non-toxic, and readily available reagent, which makes it easy to use. It could therefore be implemented routinely in dust monitoring RT-qPCR protocols. To conclude, this study is one illustration of the possible analytic improvements that will likely increase further the value of environmental sampling in HPAIV surveillance in the near future.2, rubbed on the buildings’ walls or feeders. Wipes were shipped to the National Veterinary School of Toulouse (France) and stored at 4°C for up to 48 h before being processed. Since other types of samples were not systematically collected in the 107 poultry houses included, we focused only on the dust samples in this study.Between December 2021 and April 2022, we selected a total of 107 poultry farms, either HPAI-positive as confirmed by official analyses or in close vicinity with HPAI-infected farms, in communes (administrative units) heavily affected by HPAIV epizootics. All samples were collected under the supervision of the French Official Veterinary Services and in compliance with the French legislation on notifiable diseases. In each poultry house, surface dust was collected using dry wipes of approximately 900 cmTwenty milliliter of PBS was added directly to the wipes transport bags. After mixing by hand massage for 2–3 min, the dust solution was collected and aliquoted into 1.5 mL centrifuge tubes, and stored frozen at −80°C, awaiting further analysis.7 TCID50 to 104 TCID50. Experiments were performed in biosafety level 3 laboratories.We randomly selected 9 HA and M RT-qPCR-negative dusts. We performed 10-fold serial dilutions of the virus in these dust, using the A/Mule duck/France/21348/2021(H5N1) strain previously propagated and titrated on embryonated eggs, ranging from 10Viral RNA was extracted from 100 µL of dust solution using the magnetic bead-based ID Gene Mag Fast Extraction Kit and an IDEAL-96 automated platform, according to the manufacturer’s instructions .Influenza nucleic acid load was determined by RT-qPCR, using primers and probes recommended by the WOAH, the FAO for the detection of Eurasian H5 HPAIVs and the ANSES , 18. PriThe sensitivity of the three testing protocols, for both HA and M genes, was estimated jointly through Bayesian latent class analysis (BLCA), an established paradigm endorsed by the WOAH for evaluating test accuracy when the true epidemiological status of tested individuals is unknown –31. HeN = 107) and the eight probability combinations that can be expressed as a function of the proportion of infected farms the sensitivity and the specificity of each of the three different testing protocols. In our case, given that the primers, the probe, and the templates were the same for each testing protocol, we assumed that the three testing protocols had the same specificity. To test for pairwise conditional positive dependence between testing protocols, three additional models—one for each pairwise combination  were constructed by including a covariance parameter as established in  estimates was confirmed by ensuring effective sample sizes for each parameter over 10,000 , 38. PaiModeling and analysis were performed through the JAGS library via the Rjags package version 4.12  in R ver


Preoperative anxiety is a major mental health problem during the preoperative period. Admission of women to surgery is stressful, and a high level of anxiety was associated with increased perioperative morbidity and mortality, poor treatment satisfaction, and bad obstetric outcomes, including long-term cognitive impairment in children. Despite its negative consequences, little is known on this area, particularly in the study area.To assess preoperative anxiety and associated factors among women admitted for elective obstetrics and gynecologic surgeries in public hospitals in Southern Ethiopia, 2022.th to June 20th, 2022. The Amsterdam preoperative anxiety and information scale (APAIS) was used to assess the level of anxiety. Data were collected electronically using the Open Data Kit version 2022.2.3 and analyzed with the Statistical Package for Social Sciences version 26.0. Bivariate and multivariable logistic regression analyses were done. The strength of the association was declared by using an adjusted odds ratio (AOR) with a 95% confidence interval, and a statistical significance of P < 0.05.An institution-based cross-sectional study design was conducted among 389 women using structured interviewer-administered samples who were selected by systematic random sampling from May 20The magnitude of preoperative anxiety was 57.1% (95% CI = 51.4–61.5), and 48.1% of women required an average amount of information. Being a gynecologic patient , having previous anesthesia and surgery , having fear of postoperative pain , having concern for family , having poor social support , and moderate social supports , and having a high information requirement about anesthesia and surgery  were statistically associated with preoperative anxiety.Preoperative anxiety was often high in the region. Associated factors were the type of surgery, previous anesthesia and surgery, fear of postoperative pain, fear for family, social support, and a high information need. So the national and regional health bureau should develop guidelines and implement strategies to reduce women preoperative anxiety as part of midwifery care. The women should be assessed regularly during the preoperative visits; and appropriate anxiety reduction and information regarding surgery, and anesthesia should be provided. Anxiety is an unpleasant emotional state, including tension, apprehension, uneasiness, worry, and autonomic nervous system activation, which is a personal feeling resulting from a patient's reaction to the uncertainty of particular stimuli . It is aFrom conception of their fetus to completion of childbirth via cesarean section (CS) and various gynecologic (GYN) surgeries, which involve the organs and structures of the female pelvic region, including the reproductive organ, women face numerous challenges throughout their lives . GYN surThe burden of mental illnesses such as anxiety and depression is greater than any type of chronic illness . One in Anxiety can be measured in many ways. It can be measured directly by measuring plasma cortisol and urinary catecholamine, or indirectly by measuring blood pressure and pulse . Even ifThe magnitude of preoperative anxiety among surgical patients differs in different countries' reports and operations types are as high as 92.6%  and alsoAs compared to the general population, females and patients admitted for OB and GYN surgeries suffer more preoperative anxiety than any other types of surgeries like urologic and cardiac surgeries and male patients , which aThe impact of undiagnosed and untreated preoperative anxiety among women increases the odds of perioperative complications that lead to acute or long-term impact; some of them are difficult postoperative pain management, increased hemodynamic disturbance  , 24, delA difficult issue in preoperative care is the presence of preoperative anxiety, previous published evidence suggested that there are certain factors that contribute to higher levels of pre-operative anxiety, including, low socioeconomic status, age, educational level, past surgical exposure, fear of perioperative complications as well as the fear of death from anesthesia or surgery Additionally, patients with chronic medical illnesses, who were worried about their families, and have poor social support increase the odds of preoperative anxiety , 30.There were different previous attempts to prevent and manage preoperative anxiety among surgical patients, including the establishment of preoperative educational interventions and providing non-pharmacological methods such as music therapy, chewing gum , 31, 32,Though preoperative anxiety is preventable and treatable by perioperative information provision, in southern Ethiopia, two-third (64.3%) of patients get poor quality perioperative information , and matth, 2022, to June 20th, 2022. The SNNPR is one of the eleven regions that were found in Ethiopia; its capital city is Hawassa, which is located around 273 km south of Addis Ababa, the capital city of Ethiopia. Following the separation of the two regional states (Sidama in June 2020 and the Southern West Ethiopian People's Region in 2021), the SNNPR will consist of eleven zones and six special woredas. South Regional Health Bureau reports in 2022 indicate SNNPR has 55 hospitals, 493 health centers, and 2641 health posts, for a total of 45,707 health professionals. All 55 hospitals provide maternal and child health services, including emergency obstetrics and gynecologic-related procedures with anesthesia, but only 11 of them provide major elective obstetrics and gynecologic services. It has an estimated area of 112,343.19 square kilometers, and this region has an estimated density of 132.65 people per square kilometer.This study was conducted in selected public hospitals of Southern nations’ nationalities and people’s regional states (SNNPR) from May 20An institutional-based cross-sectional study design was conducted.All women admitted for elective obstetrics and gynecologic surgeries in public hospitals in southern Ethiopia.All women admitted for elective obstetrics and gynecologic surgeries during the data collection period in selected public hospitals in southern Ethiopia.All women admitted for elective obstetric and gynecologic surgeries.Women with a known psychiatric illness or taking anxiolytic medication.The sample size for the primary objective was determined by using the single population proportion formula for a finite population with the assumption of 95% levels of confidence (Z = 1.96), 5% marginal error (d = 0.05), and the prevalence (p) of preoperative anxiety at 63% .\documen).To determine the sample size for the second objective (associated factors) that affect preoperative anxiety, it was calculated using EPI Info version 7.2 stat calculation by considering 80% power, 95% CI, and the ratio of unexposed-exposed 1 (See Table 1The sample size determined by the primary objective was greater than that of the second objective (associated factors), so the final sample size was determined by the primary objective, then added a 10% non-response rate, and the final sample size was 395.).In the southern nations, nationalities, and peoples region, there are 11 public hospitals that provide elective obstetrics and gynecologic surgeries. Among those hospitals, six were selected by a simple random sampling method (lottery). From the selected hospitals, eligible women were selected by systematic random sampling after reviewing last year's achievement registration book of elective obstetric and gynecologic surgeries that were performed in each hospital. Consecutively, every admitted woman was included in the study until the final sample size was achieved. Proportional allocation was done to allocate 395 women to each hospital , moderate (9-11), and strong (12–14) social support .The outcome variable preoperative anxiety was assessed with APAIS, which is widely used to assess pre-operative anxiety, and consists of six questions that assess three components of anxiety; anesthesia-related anxiety, surgery-related anxiety, and information desire component used and validated in different countries like Sri Lanka , and NepAPAIS was validated and translated into the Amharic version in Addis Ababa, Ethiopia; which is a reliable and acceptable tool for measuring women’s preoperative anxiety levels and their need for information about anesthesia and surgery with high reliability , and was well accepted by the study participants, with a 100% response rate with no missing value. The patient chooses the numbers that best describe the intensity of their feelings. Each APAIS item has a weight scale from 1-Not at all to 5-Extremely, the rating of five indicates the presence of high anxiety, and the sum of the scores in all items constitutes the individual’s score .The Pain was assessed with a numeric pain rating scale . Six psyPreoperative anxiety.Socio-demographic factors: age, residency, marital status, educational status, occupation, and monthly income.Social support status, Health status and clinically related factors: types of surgery, parity, chronic medical illness, previous anesthesia and surgery exposure, previous surgical complications, and pain.Possible reasons for preoperative anxiety: receiving IV fluid, fear of death, unexpected outcome of surgery, concern for family, fear of complications, cosmetics issue, fear of postoperative pain, harm from a medical mistake, need for blood transfusion, fear of the unknown, and inability to recover from anesthesia.Elective surgery( procedure): is done preplanned and performed in advance, does not involve a medical emergency, and is done by the choice of the patient or doctors .The numerical pain rating scale (NRS) is a valid method of pain assessment where patients are asked to score their pain ratings on a scale of 0–10 .Pain score- no pain (0), mild (1-3), moderate (4-6), and severe pain (7-10) .Preoperative anxiety is assessed by APAIS; it was calculated by using five Likert scale-based questions, and the scores were summed by sum scores. Those who scored 11 and above out of 20 were considered to have preoperative anxiety, whereas parents scoring below the 11 were considered to have no preoperative anxiety .High information requirement: On information scale of APAIS, patients scored 8–10 .Average information requirement: on the information scale, patients scored 5–7 .Little or no information requirement: 0n the information scale, patients scored 2–4 .www.worldbank.org/poverty.Monthly income: categorized as above the international poverty line (US$1.90) or 97.85 ETB with the current exchange rate multiplied by 30 days = 2935.5 ETB, and below poverty line: (US $1.90) or 97.85 ETB. World Bank Group. Poverty & Equity Brief: Africa, Eastern and Southern, Ethiopia, 2021. Social support is classified by sum score as poor (3–8), moderate (9–11), and strong (12–14) social support .To assure the quality of the data, a standardized and validated questionnaire was used to collect the data; a pretest was done on 10% of the sampled population; and training for data collectors was given, including theoretical and practical discussions on ODK data collection techniques. During the data collection, we reviewed the completed questionnaires for critical information before uploading them from the Android mobile phone to the ODK Toolbox server. The completeness and cleanliness of the files that the data collector sent were checked.After the data was collected through ODK, it was downloaded as an Excel file, exported to SPSS version 26, checked for completeness, cleaned, and coded. Descriptive statistics were performed to describe the study participant's characteristics. Multi-collinearity was checked by using the variance inflation factor (VIF) to determine if there was correlation between two or more independent variables, and it was tolerated when VIF < 5. The goodness of fit of the model was tested by Hosmer and Lemeshow, and it was found to be 0.17. Both bivariate and multivariable logistic analyses were used to assess the association between each independent variable and the outcome variable.p-value < 0.25 at 95% CI in the binary logistic regression model at bivariate logistic regression were transformed into the final multivariable logistic analysis in order to control all potential confounding variables. The odds ratio (OR) with 95% CI was estimated to measure the strength of association factors affecting preoperative anxiety. In this study, a variable with a p-value of < 0.05 was considered statistically significant. Finally, the data was organized and presented using text, tables, and figures based on the types of data.All variables with a ).In this study, 395 women were involved, while 389 women participated, with a response rate of 98.48%. The mean age of the study participants was 35.56 ± SD: 12.2) years, with minimum and maximum ages of 16 and 76, respectively. Among the study participants, 170 (43.7%) were within the age group of 16–31 years. Of the participants, 218 (56.0%) were urban residents. Regarding marital status, 298 (76.6%) were married, 111 (28.5%) attended secondary education, and 142 (36.5%) worked as housewives. From the study participants, 296 (76.1%) had an average monthly income above the poverty line .Of a total of 389 women who participated in this study, 203 (52.2%) were admitted for elective obstetric surgery, and 199 (51.2%) of the participants were multiparous (given 2–4 births). Concerning previous anesthesia and surgical exposures, 288 (74%) did not have any previous history of anesthesia or surgery. Of those who had previous exposure to anesthesia and surgery, 23 (22.8%) encountered previous surgery or anesthesia-related complications. 109 (28% of study participants) had a chronic medical illness, and 163 (41.9%) of participants had no preoperative pain (see Table 3).Out of 389 study participants, 159 (40.9%) of participants get poor social support, followed by 130 (33.4%) with moderate social support see Fig. .Fig. 2So.The magnitude of preoperative anxiety among women admitted for elective OB/GYN surgery in southern Ethiopia was 222 (57.1%) , with an APAIS mean (± SD) score of 12.37 ± (SD = 3.55). The minimum and maximum APAIS scores were 5 and 20, respectively. Regarding information requirements about anesthesia and surgeries, 187 (48.1%) women required an average amount of information, followed by 112 (28.8%) women who required little or no information. The mean score was 6.36 (SD = 2.12), with minimum and maximum scores of 2 and 10, respectively Among a total of 389 study participants, the most common reasons for preoperative anxiety were fear of postoperative complications 258 (66.3%), post-operative pain 257 (66.1%), concern for family 236 (60.7%), fear of death 222 (57.1%), and an unexpected result of operation 220 (56.6%). Cosmetic issues were the least common reason for preoperative anxiety in this study, accounts 100 (25.7%) See Fig. .Fig. 4ReIn binary logistic regression analysis, types of the proposed surgery, previous anesthesia and surgery exposure, chronic medical illness, fear of death, unexpected result of the operation, fear of post-operative pain, fear of post-operative complications, concern for family, social support, and information requirements about anesthesia and surgery were candidates for the final multivariable logistic regression analysis. In the multivariable analysis, type of surgery, previous anesthesia, and surgery exposure, fear of post-operative pain, concern for family, social support, and information requirements about anesthesia and surgeries were statistically associated with preoperative anxiety.The odds of preoperative anxiety among women admitted for gynecologic surgeries were two times higher than those among women admitted for obstetric surgery . Women who encountered previous anesthesia and surgery had two times higher odds of preoperative anxiety than women who had no previous exposure to anesthesia and surgeries . The odds of preoperative anxiety among women who were fear about post-operative pain were 1.9 times higher than who do not , and the odds of preoperative anxiety among women who were worried about their family were 2.5 times higher than who did not worried about their family . Women who had poor social support were 3.7 times  more likely to develop preoperative anxiety, while women who had moderate social support had 3 times higher odds of preoperative anxiety than women with strong social support . In addition, the odds of preoperative anxiety among women who needed more information about anesthesia and surgeries were 4.6 times higher than among women who required no information  . the result was in line with the study's findings of India at 55% . HoweverIn contrast, the magnitude of preoperative anxiety in this study was lower than fining from India at 63.54% ,PakistanThis study demonstrated that the odds of preoperative anxiety among women admitted for gynecologic surgeries were two times higher than women admitted for OB surgery. This is in line with study done in India . This hiThe current study found that the odds of preoperative anxiety among women who had previous exposure to anesthesia and surgery were 2 times higher than women who had no previous exposure. This finding was supported by studies conducted in Pakistan , 18. ThiIn this study, women fearing postoperative pain were significantly associated with a higher likelihood of experiencing preoperative anxiety than other women. This result was consistent with a study conducted in Gondar . This maFurthermore, women who were worried about their family were at higher odds preoperative anxiety than women who were not. This study was consistent with a study conducted in Ethiopia . This maThe odds of having preoperative anxiety are higher among women who had poor and moderate social support as compared to women having strong social support. The results of this study were supported by other studies conducted in Egypt, and Colombia , 44. ThiThis study suggested that the odds of preoperative anxiety among women who need more information about anesthesia and surgeries were 4.6 times higher than women who required no information. This finding was supported by studies conducted in Addis Ababa, and Nepal Similar to studies conducted in Addis Ababa , 44. ThiThere may be social desirability bias, and the level of anxiety may differ based on specific times in the preoperative period; hence, it was not addressed. Also, because the study design is cross-sectional, it was difficult to establish a cause-and-effect relationship.Preoperative anxiety was often high in the region. Associated factors were the type of surgery, previous anaesthesia and surgery, fear of postoperative pain, fear for family, social support, and a high information need. So the national and regional health bureaus should develop guidelines and implement strategies to reduce women's preoperative anxiety as part of midwifery care. The women should be assessed regularly during the preoperative visits, and appropriate anxiety reduction and information regarding surgery and anaesthesia should be provided. Further researcher would be responsible for assessing coping strategies to reduce and prevent preoperative anxiety.


Rotary friction welding (RFW) is a green manufacturing technology with environmental pollution in the field of joining methods. In practice, the welding quality of the friction-welded parts was affected by the peak temperature in the weld joint during the RFW of dissimilar plastic rods. In industry, polycarbonate (PC) and acrylonitrile butadiene styrene (ABS) are two commonly used plastics in consumer products. In this study, the COMSOL multiphysics software was employed to estimate the peak temperature in the weld joint during the RFW of PC and ABS rods. After RFW, the mechanical performance and microstructural evolution of friction-welded parts were investigated experimentally. The average Shore A surface hardness, flexural strength, and impact energy are directly proportional to the rotation speed of the RFW. The quality of RFW is excellent, since the welding strength in the weld joint is better than that of the ABS base materials. The fracture occurs in the ABS rods since their brittleness is higher than that of the PC rods. The average percentage error of predicting the peak temperature using COMSOL software using a mesh element count of 875,688 for five different rotation speeds is about 16.6%. The differential scanning calorimetry curve for the friction-welded parts welded at a rotation speed of 1350 rpm shows an endothermic peak between 400 to 440 °C and an exothermic peak between 600 to 700 °C, showing that the friction-welded parts have better mechanical properties. Rotary friction welding (RFW)  is one oEslami et al.  reviewedPolymers are frequently used in some structures, because the major difference between metal and polymers is that polymers are lighter than metal. Acrylonitrile butadiene styrene (ABS)  and polyS 7, 8, 9, and 12 [According to the research results, the remarkable findings provide potential industrial values in the polymer-welding industry, since the RFW of dissimilar polymer rods is a green manufacturing method based on four sustainable development goals, i.e., SDG, and 12 ,35,36,37, and 12 ,39. In t, and 12 ,41 are r, and 12 ,43,44,45The average surface hardness, flexural strength, and impact energy in the weld joint are increased with increasing rotation speed of RFW. The maximum joint strength of 132 MPa, Shore A hardness of 80, and impact energy of 156 J are obtained at a rotation speed of 1350 rpm.The quality of RFW is excellent, since the welding strength in the weld joint is better than that of the ABS base materials. The fracture occurs in the ABS rods since the brittleness of the ABS rods is higher than that of the PC rods.The average percentage error for predicting the peak temperature using COMSOL software using a mesh element count of 875,688 for five different rotation speeds is about 16.6%.The heat capacities for the friction-welded parts welded by rotation speeds of 330 rpm, 490 rpm, 650 rpm, 950 rpm, and 1350 rpm are −1.002 mW/mg, −0.8127 mW/mg, −0.626 4mW/mg, −1.759 mW/mg, and −2.287 mW/mg, respectively. The DSC curve for the friction-welded parts welded by a rotation speed of 1350 rpm shows an endothermic peak between 400 to 440 °C and an exothermic peak between 600 to 700 °C, showing that the friction-welded parts have better mechanical properties.The advantages of RFW include efficiency of production, low heat input, and environmental friendliness. PC plastic has very good heat resistance and is widely employed for more robust materials. ABS plastic provides high tensile strength and is very resistant to chemical corrosion and physical impacts. To establish domain knowledge of the RFW of ABS and PC rods, this work reports the joining of FDM-printed dissimilar thermoplastic PC and ABS rods. To establish domain knowledge of the RFW of ABS and PC rods, the RFW experiment was performed using a turning machine. The main conclusions from the experimental work in this study are as follows:


BackgroundImmunizations protect children from deadly infectious diseases. The timeliness of vaccinating children is crucial to ensure effective immunization and to decrease the burden of many infectious diseases. Therefore, this study assessed the prevalence and determinants of vaccination delay among children in Riyadh City, Saudi Arabia.MethodsThis cross-sectional study was conducted at the primary healthcare centers in Riyadh City, Saudi Arabia, on 593 parents with children of two years of age or below. It used a self-administered questionnaire inquiring about socio-demographic characteristics and assessing the vaccination statuses of their children and the causes of delayed vaccinations.ResultsThe results showed that 7.1% of children had a delay in the previous vaccination. Of those delays, collectively, 77.5% were delays in inactivated poliovirus vaccine (IPV), oral poliovirus vaccine (OPV), and meningococcal vaccine (MCV) vaccines. The delay was mostly caused by an illness of the child on vaccination day, carelessness of parents, or long postponement. After adjusting for confounders, the father's high school or bachelor's education level  (p<0.05), child's mix-type nutrition  (p=0.001), and the belief that multiple vaccines are harmful to the child  (p=0.005) were positively associated with vaccination delay, while prematurity was negatively associated with vaccination delay  (p=0.031).ConclusionThe study found the prevalence of vaccination delay was lower than in previous COVID-19-era studies. The child's illness was the main reason for the delay. Factors like parental education, nutrition type, and vaccine beliefs contributed to delays, while prematurity reduced delays. Measures should be strengthened to increase vaccination coverage for children. It is estimated that over 2.5 million deaths are avoided through vaccination every year, and according to the WHO, around 19.5 million infants worldwide, unfortunately, do not get basic vaccines  despite Haemophilus influenzae type b (HIB) vaccines, BCG, meningococcal vaccine, varicella, MMR, and hepatitis A [In the Kingdom of Saudi Arabia, immunization started with diphtheria, tetanus, and pertussis (DTP) vaccination in 1979, and it was later expanded to include additional vaccines. Vaccination is considered a prerequisite for school entry at the age of six years in Saudi Arabia . The Sauatitis A . Vaccinaatitis A ,5, couldatitis A ,7.Vaccine acceptance and practice are affected by factors such as being unwilling to vaccinate among well-educated upper and middle-income people, which is related to health beliefs . SocioecThe COVID-19 pandemic has also affected immunization systems and schedules worldwide . Three-qThese reported challenges highlight the urgent need to explore further the vaccination program in Saudi Arabia to inform policies and programs designed to get back on track and ensure full coverage for all children in Saudi Arabia. Post-pandemic studies are essential to evaluate the vaccination delays and contributing factors in order to inform health authorities, public health professionals, and partners in Saudi Arabia in their efforts to streamline successful vaccination strategies. Therefore, we conducted this post-pandemic study to assess the prevalence and determinants of vaccination delay in children aged two years or less in Riyadh City, Saudi Arabia.Study design and settingWe conducted an analytical cross-sectional study between January and March 2023. We targeted Saudi parents of healthy children aged two years or less attending the PHCCs and vaccination in Riyadh City who had vaccination cards.Parents who were non-Saudi, who were not accompanied by their children, and who had sick children were excluded.Sample size and sampling methodAssuming a 95% confidence interval, 5% margin of error, and response distribution of 50% with a design effect of 1.5, the estimated minimum sample size was 566. By using a convenient sampling technique, we selected 593 eligible participants from PHCCs.Data collection tool and techniqueData collection was done using a self-administered questionnaire in Arabic language. The questionnaire consisted of two parts: the first part included child characteristics, parental information, the child’s physical well-being, socioeconomic status, number of delayed vaccinations, duration and reasons for vaccine delays, and the second part assessed parents’ awareness. Data on vaccination timing were collected from vaccination cards.The questionnaire was prepared by the authors, reviewed for validity by three consultants in family and community medicine, and then pre-tested for comprehension on 15 participants who were not included in the study. The sample has been taken from Riyadh's second health cluster, which has 40 PHCCs; a random sample of 10 PHCCs has been selected with an average of 60 participants from each center. Then, the investigators visited each PHCC and invited the parents to participate in the study during the waiting time after explaining the objectives of the study to them. The questionnaire sheets were then handed over to the parents, who consented to fill them, and they were immediately collected afterward.Data management and statistical analysisData entry and analysis were done using SPSS Statistics version 23 . We performed descriptive statistics and presented data as frequencies, percentages, or median (IQR). Regression analyses were performed to identify associations between the prevalence of vaccination delays and socio-demographic characteristics. The OR and CI were calculated, and a statistical significance was considered when the p-value was less than 0.05.Ethical considerationsEvery participant was informed about the purpose of the study, written consent to participate was obtained, and they were allowed to withdraw at any time. The questionnaire was anonymous to ensure confidentiality, and no private questions were included. The data collected were never disclosed to anyone outside this study. Ethical approval was obtained from the King Fahad Medical City Institutional Review Board (Ref. No: 23-050E).The majority of participants were aged 26-35 (53%), followed by 36-45 years old (25%). Most participants had 18- to 24-month-old children (20%), followed by 9 to 12 months old (18%). Regarding the sex of the children, it was found that most of them were female (53%), and the rest were male. The majority of mothers had bachelor's degrees (44%), followed by those with high school certificates (39%), and most mothers were employed (57%). The majority of fathers were aged 36-45 (54%), followed by those who were 26-35 years old (33%). Like mothers, most fathers had bachelor’s degrees (61%), followed by high school certificates (22%), and 97% of fathers were employed. Most families had an income of more than 10,000 Saudi Rials (SR) (46%), followed by families with 5000-10,000 SR income (32%), and 61% of the families had two to four children. Table As shown in Table As seen in Table Regarding vaccination awareness, 58.3% of participants know that they prevent infections, 18.6% think vaccination is just for following protocols, and 18.6% vaccinate to complete the table to attend school. Some participants believed there was a link between the MMR vaccine and autism (15.5%), while 39.5% did not know. Most side effects reported were fever 44.4%), autism (7.4%), pain (3.7%.), and rash (3.7%.). However, 33.3% could not identify any side effects. Most (66.9%) thought that children should be vaccinated as scheduled. When the vaccine is unavailable, most participants reported trying to find it in another center or clinic (60.7%), followed by those who returned a month later (23.6%). Within 48 hours post-vaccination, most participants (41%) reported giving paracetamol to children with fever, while others used cold compression (32.6%) and called doctors to ask (18.8%). Over half (53.1%) of participants did not know the impact of the COVID-19 pandemic on their decision, and 29.5% thought that giving multiple vaccines is harmful to the child. Table 4.4%, autThe univariate regression analysis showed that parents’ education, family income, child’s gestational age at delivery, child’s current nutritional intake, and parent’s belief that giving multiple vaccines was harmful to the child were significantly associated p<0.05) with vaccination delay , children receiving mix-type of nutrition , and parents thinking that giving multiple vaccines was harmful to the child  were positively associated with vaccination delay, whereas a child born prematurely  was negatively associated with vaccination delay  of delayed vaccines reported in Saudi Arabia by previous studies ,31. ThisThere are some limitations to the study. For instance, using a convenience sample limits the generalizability of the findings to the entire population. Moreover, using a cross-sectional study design does not permit concluding the causality of the emergent predictive relationship, and the self-reported questionnaire is prone to over- and under-reporting that may affect the interpretation of results. This study was also conducted in a single city, which might not reflect the situation in other cities and rural areas of Saudi Arabia. We recommend more extensive studies encompassing nationwide samples with prospective designs to mitigate these limitations and show vaccination coverage trends in the future.The findings showed that the prevalence of vaccination delay was lower than what previous studies reported, most of which were conducted during the COVID-19 pandemic, indicating an improvement in vaccination coverage, especially post-pandemic. The most common reason for the delay was reported to be a child’s illness. However, the significant independent determinants, such as parental level of education, child’s nutrition type, and parent thinking that giving multiple vaccines was harmful to the child, contributed to more vaccination delays, while prematurity reduced the likelihood of delays. Therefore, the study's results show that vaccine delay prevalence was low, but measures were strengthened to keep increasing the coverage for all children.


Mycobacterium bovis, the causative agent of bovine  and zoonotic tuberculosis, it can be challenging to discriminate between these two states. This is a result of the combination of suboptimal detection tests so that the actual extent of the pathogen prevalence is often unknown, as well as of the low genetic diversity, which can hide the temporal signal provided by the accumulation of mutations in the bacterial DNA. In recent years, the increased availability, efficiency, and reliability of genomic reading techniques, such as whole-genome sequencing (WGS), have significantly increased the amount of information we can use to study infectious diseases, and therefore, it has improved the precision of epidemiological inferences for pathogens such as M. bovis. In this study, we use WGS to gain insights into the epidemiology of M. bovis in Cameroon, a developing country where the pathogen has been reported for decades. A total of 91 high-quality sequences were obtained from tissue samples collected in four abattoirs, 64 of which were with complete metadata. We combined these with environmental, demographic, ecological, and cattle movement data to generate inferences using phylodynamic models. Our findings suggest M. bovis in Cameroon is slowly expanding its epidemiological range over time; therefore, endemic stability is unlikely. This suggests that animal movement plays an important role in transmission. The simultaneous prevalence of M. bovis in co-located cattle and humans highlights the risk of such transmission being zoonotic. Therefore, using genomic tools as part of surveillance would vastly improve our understanding of disease ecology and control strategies.When studying the dynamics of a pathogen in a host population, one crucial question is whether it transitioned from an epidemic  to an endemic stable state . For slow-growing and slow-evolving clonal pathogens such as  In the last two decades, the increased availability, efficiency, and reliability of genomic reading techniques, such as whole-genome sequencing (WGS) techniques, have ignited a profound transformation in understanding disease ecology and epidemiology. This, coupled with improved statistical methodologies and high-performance computing, has enhanced our understanding of pathogen dynamics and evolution .Techniques such as WGS can identify polymorphisms in the genetic material, which is generated by transcription errors that can occur to the pathogen while replicating within their host . As the Mycobacterium tuberculosis Complex (MTBC) members are clonal species, and therefore, recombination has been considered rare . The Krraken v2   with Burrows-Wheeler Aligner v0.7.17 (SAMtools v1.10 (M. tuberculosis were determined by querying the sequences using NCBI web nucleotide BLAST with the default parameters (Integrative Genomics Viewer v2.14.1 (We checked if divergent sequences belonged to other mycobacteria species. We tested the presence of regions of difference (RDs) 1, 4, 9, and 12 patterns  in the o v0.7.17  and sortls v1.10 . Primer rameters , while t v2.14.1 .M. bovis genomes from online repositories as follows: first, from the Patric  dataset , which included all the Cameroonian high-quality sequences (n = 91) and the other African ones plus the 1997 reference from the UK (n = 121).When analyzing sequences from b server , 41 to cn = 64) of the tree using the TN93 genetic distance model and the neighbor-joining (NJ) algorithm ape package  normally distributed clock rate prior, with mean and standard deviation set as the slope in the root-to-tip obtained in Tempest; the chain length was set to 108 and sampled every 104 steps. The models were compared using marginal likelihood estimation (MLE), with path sampling (PS) and stepping-stone sampling (SS), if they reached a satisfactory effective sample size (>200). Once the model features were selected, we ran a final one setting the chain length to 109 steps and sampled every 105 steps. In this case, we used the clock rate posterior of the selected exploratory model as a prior for the final model. The maximum clade credibility (MCC) tree was extracted with TreeAnnotator v1.10.4 (part of the BEAST suite), and clades were visually defined within the MCC tree branches. The MCC tree was plotted against the sequence spoligotype and MIRU-VNTR typing to visually assess the correspondence between molecular typing and clades.To select the best model, we ran a series of exploratory models using an HKY  substituSkygrid , 52 for BEAST run, we extracted a set of 100 trees from the posterior distribution and further analyzed using seraphim v1.0 , and four land cover types . The original raster layers were downloaded from online repositories , calculated as BFe = pe/(1 – pe), where pe is the probability that Qobserved > Qrandomized. We used two criteria for tree randomizations as follows: (1) randomizations of node positions while maintaining the branches' lengths, the tree topology, and the location of the most ancestral node; and (2) randomizations of node positions while maintaining only the branches' lengths.Once we identified the potential resistance or conductance factors, we performed 10 tree randomizations and calculated the statistics again. In this case, we used the Bayes Factor  and unweighted . For this network, we computed six variables to be associated with each pair of isolates: degree and betweenness centrality  of both R package igraph  on the test dataset. These were both calculated using the package caret , with 91 having enough read depth and genome coverage to allow further analyses  belonged to Af1 clonal complex, and except for one, the spoligotypes were already known; for the other, we identified a new pattern (hex code: 6F-1F-5F-7F-BF-40). Being characterized by the absence of spacer 30, this spoligotype was considered Af1 (n = 32/89).We analyzed 124 ered Af1 . The domwww.Mbovis.org by Egbe et al.  was slightly higher than the average distance of the 1997 UK Reference from the Cameroonian isolates (222 SNPs), and they did not cluster with any other WGS sequence sampled in Africa . For bote et al. . Followie et al. , we testin Spain  . For two cattle (one from Bibemi and the other from Touboro), two M. bovis isolates sequenced were available (obtained from different tissues). In both cases, the two strains were identical  and a slope of 1.267 × 10−2 , while the average clock rate was 1.32 × 10−7 substitution/site/year (95th HPD 1.20 × 10−7 – 1.44 × 10−7). The maximum clade credibility (MCC) tree is presented in M. bovis isolates per administrative subdivision, which ranged from 1 to 17  as follows: mosaic, shrub, and other vegetation covers (with both path models as resistance in the least-cost one); forest cover, elevation, and waterbodies cover ; and cattle density (as resistance). However, when their statistical significance was tested through randomization, only forest cover and elevation (both as conductance) showed a Bayes factor significance .The estimated MCC tree located at the most recent common ancestor (MRCA) in Touboro (North region), and from there, a rapid expansion of the outbreak reached most of the study area by the early 1970s. From the estimated origin, the pathogen likely spread first northward to Garoua (in the same region) and westward to the Northwest region, and later to the Extreme North and Adamawa regions and again to the Northwest.M. bovis is known to be quite effective in spreading at the wildlife–livestock (and humans) interface  were retained. Conversely, only five network-related variables were retained, three for the cattle movement network (out of eight) and two for the spatial network (out of six). All network-related variables had a positive effect on the SNP distance, with the number of cattle moved in or out of a subdivision , having the higher predictive effect. Interestingly, this result was similar to other studies where cattle movements alone could not fully capture M. bovis genetic diversity . Generally, the pipelines used to call the SNPs differed in many of the aforementioned studies, contributing to the uncertainty of the estimates and potentially generating biases in the analysis results and the clock rate calculations.In agreement with many studies and with the e genome , 80, 81.u et al.  used M. M. bovis is unlikely in North Cameroon, but rather the disease is slowly expanding over time. Our findings highlight the importance of collecting data in underrepresented areas to enrich insights into the current body of literature, predominantly from developed countries. Moreover, our results pave the way for future research aimed to understand whether the observed M. bovis high-genetic diversity affects the spread dynamics.In conclusion, our study indicates endemic stability of M. bovis control  at EMBL-EBI under accession number PRJEB61415.BB, LN, and VT conceived the original project. BB, RK, LN, VT, MS, and NE designed the field study, databases, and survey instrument. RK, NE, VT, and BB developed the field SOPs and collected the data. PM collected the cattle movement data. GR, AM, FD, and FE cleaned the data. GR, BB, SL, and AM conceived the quantitative analysis. GR and SL run the phylogenetic analysis. BS implemented the bioinformatical pipelines. GR performed the machine learning analysis and responsible for writing the initial drafts. All authors contributed to comments for the final draft, contributed to the article, and approved the submitted version.