To evaluate factors associated with the performance of children andadolescents with cystic fibrosis (CF) in the Modified Shuttle Test (MST) andcompare it with healthy children and adolescents.2). Statistical analysis included Mann Whitney andSpearman coefficient tests, being significant p<0.05. This is a cross-sectional study, with children and adolescents divided intotwo groups: cystic fibrosis (CFG) and control (CG). Variables evaluated inthe MST: walking distance, test level, heart rate variation (∆Hr), post-testmean arterial pressure (MAP Pt) and peripheral oxygen saturation variation(∆SPO2B) and level test. The ∆Hr and MAP Pt had amoderate positive correlation with distance and level test for both groups. In CFG, the leveltest had a significant association (r=0.4 - p=0.02) with%FEV1. Sixty individuals aged 6-16 years old were evaluated. Anthropometric datawas similar between groups. Differences between groups were shown for:baseline heart rate (BHr), peak heart rate (PHr), ∆Hr, recovery heart rate(RHr), post-test respiratory rate (PtBr), saturation variables, peripheraloxygen level (SpO Children with cystic fibrosis presented functional limitation in theModified Shuttle Test, which was influenced by lung function. The exercise capacity can be assessed through laboratory and field tests,such as the modified shuttle test (MST) ,,Several studies were conducted with the Shuttle test with 12 levels to follow up thepopulation with CF;Cross-sectional study approved by the Research Ethics Committee of PontifíciaUniversidade Católica de Minas Gerais  and by the Committee of Fundação Hospitalar doEstado de Minas Gerais (FHEMIG) (1.753.013).Children and adolescents with CF who did not present with pulmonary exacerbation wereselected and followed up at Hospital Infantil João Paulo II (HIJPII), in FHEMIG, inBelo Horizonte, besides healthy children and adolescents attending public schools.Participants were paired according to sex and age. The study began after the consentand assent forms were signed by tutors and participants, respectively. The exclusioncriteria were considered in the exercise test for both groups: hemodynamicinstability, significant changes in heart rate and blood pressure, orthopedic,neurological or rheumatologic disorders. In the group of healthy children, those whopresented with respiratory disorders, pointed out by the International Study ofAsthma and Allergies in Childhood (ISAAC), were excluded. ISAAC is validated,reproducible and easy to apply, translated to Brazilian Portuguese.2).Two variables of response were defined: walking distance, test level, and both referto functional performance. The following were mentioned as co-variables: heart ratevariation (∆Hr), post-test mean blood pressure (Pt MBP) and peripheral oxygensaturation (∆SpO1), forced vitalcapacity (FVC) and FEV1/FVC were the studied variables, obtainedaccording to the maximal expiratory flow-volume curves using a Jaeger FlowProspirometer . The results were presented in percentagenumbers, according to the equations of reference.Pulmonary function was assessed in both groups, according to the guidelines of theAmerican Thoracic Society (ATS).2) and Hr were constantly measuredduring the test, and the respiratory rate (Rr), blood pressure (BP) and modifiedBorg scaleThe adapted protocol of 15 levels of the Shuttle test, described by Bradley etal.,2 was measured by pulse oximetry .During the test, the verification of Hr was carried out with a heart sensor, whichsent the data to a wrist watch .SpOSample was calculated to compare the groups using the software GPower 3.1, whichconsidered power of 90%, alpha error of 0.05 and effective size of 0.80. Thus,sample size in each group was estimated in 28 subjects.Statistical analysis was performed using the Statistical Package for the SocialSciences (SPSS), version 22.0. The normality of data was assessed by theShapiro-Wilk test, and demonstrated asymmetric distribution. Therefore, data wereexpressed in median and interquartile range, and non-parametric tests were used. TheMann-Whitney test was used for the comparison between groups. For the correlation ofMST performance variables and co-variables, the Spearman coefficient was used. N allanalyses, the considered significance was 0.05.Staphylococcus aureus, in 70% of the patients  and CF group (CFG). The median age of participants was 10.5years, of which 60% were male. In the CFG, 23.5% were classified as eutrophic; 60%,malnourished (low weight); and 16.5% were overweight. In the CG, 30% were eutrophic;43.3% presented with low weight; and 26.7% were overweight. There was no statisticaldifference between the assessed groups regarding anthropometric data. In CFG, themost common genetic mutation and bacterial colonization were: 508del heterozygote,in 53%, and patients . Lung di33.5%: mild.30%: moderate.10%: severe.26,5%: did not present with lung disease.The Hr behavior in the test is presented in 2B) and advanced level ,peripheral basal oxygen saturation . In the CG, thelower the saturation variation in the test, the higher the functional performance.The correlations between the performance variables and the studied co-variables arein 1 predictor in theCFG and the CG , respectively. The correlationsbetween the percentage of the forced vital capacity (%FVC) predictor and theresponse variables in both groups were moderate, positive and significant, both fordistance (CFG and CG: r=0.4 and p=0.02) and reached level .In both groups, the association between the performance variables and the pulmonaryfunction of participants was also assessed. The reached level presented moderatepositive correlations with the percentage of the FEVr, Pt MBP and ∆SpO2 in bothgroups. Besides, int he CFG pulmonary function was closely related to theperformance in the test. The impact of lung disease relates to the functionalworsening in the MST of children with CF, resulting in a shorter distance whencompared to healthy individuals.The main findings of this study showed that the factors associated with the bestperformance in the test were ∆HThis study pointed out that MST caused the Hr to increase in both groups. We appliedthe Shuttle test with 15 levels, in which the participant can run, besides walking.This fact may have contributed with the higher increase in Hr, as observed by Lanzaet al.,One criterion addressed at assessing the physiological responses of thecardiopulmonary system to exercise is Hr behaviorFlorêncio et al.Ketchell et al.,Efficient Hr recovery is considered as a marker of physical aptitude and favorableprognosis in the follow-up of patients with CF.Mean BP is an indirect measurement that includes systolic and diastolic BP. The BP ofchildren and adolescents behaves similarly to that of adults; values areproportional to age and intensity of the exercise.2, this variable also presented an association withperformance in both groups. In the CFG, the better the performance assessed usingthe walked distance, the higher the oxygen desaturation, which demonstrates that theoverload imposed to the respiratory system had a direct interference on saturationreduction. The O2 desaturation in exercise in the population with CF mayoccur because of the damaged pulmonary function triggered by the condition. Theincreased effort requires higher ventilatory response; however, because of thecompromised lung capacity, the gas exchange is inefficient, so there isincompatibility in the ventilation-perfusion ratio, and reduction in gasdiffusion.2 during exercise in patients with CF. Thiscan be explained because this test represents the activities of daily living, andmore muscle mass involved in walking or running. Other authors mention that SpO2 is an important variable to determine the clinical condition ofthe patient with CF, because the Hr and SpO2 response is associated withthe severity of the disease.Regarding SpO,,Concerning the walked distance, the CFG presented with worse performance incomparison to the CG. This situation may have been owed to the limitation ofexercise, which is common in this population.The reduction in pulmonary function is one of the main consequences of thecomorbidities caused by CF. In this study, we observed that in the CFG, the higherthe values of FEV1 and FVC, the higher the walked distance in the MST. The reducedpulmonary function had a direct impact on functional ability, assessed with the MST.Previous studies with the population of patients with CF showed similarresults.The proper weight and normal muscle mass are directly related to adequate growth andgood pulmonary function in patients with CF. Lung disease can often be related tonutritional decline.The level of pulmonary dysfunction in patients with CF is a limitation of this study,since 60% of them had mild or no dysfunction. Therefore, a population whose profilewas stratified by the level of lung dysfunction could have increased the inferenceof the associations. Besides, due to its cross-sectional design, the study does notallow the establishment of a cause-and-effect ratio; for that, a longitudinal cohortshould be analyzed.The conclusion was that patients with CF present with higher functional and cardiaclimitation, assessed by the MST, in comparison to healthy individuals, and thatfactors associated with performance were similar in both groups. The anthropometricvariables did not interfere in the performance of the groups, and cardiopulmonaryfunction directly influence the functional capacity of individuals with CF.


Meta-analysis is a statistical process summarizing comparable data from a number of scientific papers. The use of meta-analysis in microbiology allows decision-making that has an impact on public health policy. It can happen that the primary researches come to different conclusions, although these are targeted with the same research question. It is, therefore, inevitable to have the means to systematically evaluate information and compare research results. Ulcerative colitis together with Crohn’s disease are among the two main inflammatory bowel diseases. This chronic disease of the gastrointestinal tract, with an as yet unclear etiology, is presented by an uncontrolled inflammatory immune response in genetically predisposed individuals to as yet undefined environmental factors in interaction with the intestinal microbiota itself. In patients with ulcerative colitis (UC), changes in the composition and relative abundance of microorganisms could be observed. Sulfate-reducing bacteria (SRB), which commonly occur in the large intestine as part of the commensal microbiota of animals and humans involved in the pathogenesis of the disease, have been shown to occur. SRB are anaerobic organisms affecting short-chain fatty acid metabolism. This work outlines the perspectives of the use of meta-analysis for UC and changes in the representation of intestinal organisms in these patients. Ulcerative colitis (UC) is one of the chronic inflammatory diseases of the gastrointestinal tract ,2. Its e2S) has been confirmed in patients with active UC, and this fact has been linked to an excess of SRB in the literature [A low number of protective bacteria that produce short-chain fatty acids can lead to increased mucosal permeability. Conversely, enormous concentrations of some bacteria can increase the production of toxic metabolites that increase mucosal permeability and block butyrate metabolism . The larterature .Meta-analysis is a statistical process summarizing comparable data from a large number of scientific papers. It is a quantitative and formal statistical technique that is used for the systematic evaluation of the results of previous research to reach valid conclusions ,19,20. DThe aims of the research study were to analyze existing information on ulcerative colitis, possible factors influencing the disease, its spread and connection to nutrition, description of the association of intestinal SRB with the development of this idiopathic intestinal inflammation and intestinal microbiota changes as well as to describe a meta-analysis as a method for evaluating these processes and comparing in intestinal microbiota perspectives of using meta-analysis for UC. The main points of the review are shown in Ulcerative colitis together with Crohn’s disease are the two main inflammatory bowel diseases (IBD) ,23,24,25There are significant differences between the two diseases. Crohn’s disease can affect any part of the gastrointestinal tract, from the oral cavity to the rectum. It most often affects the distal part of the small intestine. UC causes inflammatory changes only in the colon and rectum . UC is mHistological findings in patients with UC are characterized by the presence of inflammation, which causes the destruction of mucosal cells. Atrophy of Lieberkühn’s crypts and loss of cryptic architecture are present at all stages of the disease. Neutrophil infiltrations, goblet cell depletion, and crypt abscesses are significant in the active UC. The latter symptom can also be found on the CD . The diaPathological findings associated with UC include increased some inflammatory mediators, signs of oxidative stress, impaired microbiome, abnormal glycosaminoglycan (GAG) content in the mucosa, decreased oxidation of short-chain fatty acids (SCFAs), increased intestinal permeability, increased sulfide production, and decreased methylation. Although the primary trigger for UC has not been identified, numerous sub-processes have been elucidated. The interconnection of these findings could lead to clarification of the etiology of the disease described above .Significant ethnic and geographical differences in incidence and prevalence, the consensus in twins, recurrence in families, and association with genetic syndromes have reinforced the importance of genetics in UC .However, the genetic mechanism involved in the development of the disease has not been fully elucidated. It does not include a single dominant gene or a single recessive gene with full penetration. The most probable mechanism appears to be the combined interaction of several genes, the so-called polygenic or rather multifactorial type of inheritance. Indeed, studies suggest that non-genetic factors  are able to trigger UC in a genetically susceptible host .UC, like other complex diseases, results from the common effects of polygenic variations . One of NOD2, which is the strongest causal gene for CD production but also shows significant protective effects in the pathogenesis of UC [Risk effects specific for UC only were confirmed in 23 loci and the remaining 30 for CD only. Individual gene variants at many of these loci have not yet been investigated, and even less is known about their functional implications. Interestingly, 43 of these 53 disease-specific loci show the same direction of action in both CD and UC, suggesting that almost all biological mechanisms involved in one disease play a role in the other . One intis of UC ,33.Helicobacter pylori infection) later in life [Several theories have been proposed to explain unknown environmental exposures that may interact with the immune system and lead to an abnormal inflammatory response to the intestinal microbiome. The most important theory suggests that the increasing frequency of immunological disorders can be attributed to excessive hygiene in childhood and insufficient contact of children with intestinal pathogens. These children are more likely to develop an inappropriate immune response to exposure to new antigens  has been observed. The reduction of the genus Bifidobacterium was also observed in the intestinal contents. In contrast, the proportion of Proteobacteria (including SRB) and Actinobacteria is increasing in patients with UC. Significant differences were found in the number of bacteria that penetrated the mucosal barrier into the intestinal mucosa between patients with UC and healthy individuals [Several researchers have confirmed the existence of so-called dysbiosis of the microbial intestinal microbiome in patients with UC. This term refers to changes in the composition and representation of individual species of microorganisms in comparison with healthy individuals. A marked reduction in the incidence of ividuals ,22. Someividuals ,40. The The unresolved question is whether chronic, recurrent inflammation is caused by persistent infection with a specific pathogen, excessive exposure to normal luminal bacterial products due to increased intestinal permeability, or an abnormally aggressive immune response to luminal components .α within the lamina propria of the intestinal wall. IFN-γ is also produced and increases intestinal permeability. The production of IL-2, IL-10, TNF-β, and transforming growth factor-beta, which regulates cytokinin levels in patients with IBD, is reduced, and this may explain chronic inflammation in patients [The main mechanism of inflammatory damage in patients with UC is mediated by an abnormal or exaggerated immune response. HLA (human leucocyte antigen) class II molecules, which mediate autoimmune damage, are found in large numbers within the intestinal epithelial cells of patients with active IBD. HLA class II molecules are responsible for antigen processing and presentation . Activatpatients .In the absence or after the functional loss of these anti-inflammatory feedback signals, physiological defense mechanisms may turn into pathological responses. The data suggest that disorders in homeostasis between signals obtained from bacteria and hosts at the epithelial cell level lead to disruption of intestinal barrier function and thus the development of mucosal immune disorders in genetically sensitive hosts .UC has an annual incidence of 10–20 per 100,000, but these data are generally considered underestimated. It mainly affects the younger population with a maximum incidence between 20 and 40 years but can affect any age group and up to 15% of individuals are over 60 years of age at the time of diagnosis. It is currently estimated that IBD affects up to 1.4 million people in the United States and 2.2 million in Europe. Gender differences have been observed in patients with a slight predominance of men . UlceratThe incidence of this disease in the Czech Republic is between 12–15 patients per 100,000 population, i.e., the mean value between Western and Eastern Europe. However, the available data show a high increase in the total number of patients treated. In the case of the Czech Republic, the number of outpatients increased from about 20,000 in 2007 to 24,000 in 2013. There was also an increase in the number of newly diagnosed cases from the number of newly diagnosed cases in the monitored period 1700 to 2500. The number of deaths with the main diagnosis of UC in the Czech Republic varies from 26 to 50 cases during each year [The large intestine (intestinum crassum) is the last part of the digestive tract ensuring the resorption of water, amino acids, bile acids, salts, vitamins and removes unabsorbed residues such as feces. This organ is 1.3–1.4 m long and 5–8 cm wide. The main sections of the large intestine are the caecum from which the appendix vermiformis, the colon, the rectum, and the anal canal analis extend. The intestinal wall consists of mucosa, mucosal ligament, muscle, and serosis.The mucosa of the large intestine is smooth, slightly articulated. Kerckring’s algae and intestinal villi, which are found in the small intestine, are absent in the large intestine. Lieberkühn’s crypts have a tubular appearance and are significantly longer than in the small intestine. The epithelium of the colonic mucosa is single-layered cylindrical and consists of enterocytes and goblet cells, which are particularly abundant in the middle sections of Lieberkühn’s crypts. Paneth cells are usually missing. Of the endocrine cells, the epithelium contains mainly EC cells , which, among other things, synthesize serotonin. The intestinal epithelium is a physical barrier that coexists with intestinal microorganisms, ensuring the transport of substances and their regulation so that homeostasis is maintained .13–1014 bacteria in the human gut, with about 1013 cells in the human body [There are approximately 10man body ,47. Thusman body ,39. Bactman body .Bacteroidetes, Firmicutes, Actinobacteria, Proteobacteria, Verrucomicrobia, and also representatives of the domain Archaea predominate. Bacterial strains of Cyanobacteria, Fusobacteria and Spirochaete are also present in the intestines, but in smaller numbers. The microbiota has an important metabolic, immunological, and protective function [Profiling of the 16S rRNA gene sequence showed that bacteria of the strains function .The composition of the microbiome is therefore crucial because billions of bacterial individuals in the intestinal lumen can become a threat to the host organism with any change in conditions. Although metabolites produced by the microbiota affect various organs and systems in the body by signaling, symbiosis with the intestinal mucosa is necessary for their survival . The maiThe human body has well-developed mechanisms by which it effectively prevents the translocation of symbiotic or pathogenic microorganisms across the mucosal barrier, including mucus protection of the intestinal epithelium. Intestinal mucus has a highly organized glycoprotein network structure but also contains a stable proteome . IntestiAnaerobic colon bacteria break down carbohydrates and proteins through fermentation into gases and SCFAs , which aIndigestible polysaccharides present in, for example, fruits and vegetables pass without effective cleavage to the large intestine, where they are fermented to form SCFAs as products of bacterial metabolism . Low int4, NH3, H2, CO2, and H2S [Bacteroides and Propionibacterium are the dominant proteolytic species in fecal samples, with proteolysis also being common to Clostridia, Streptococci, Staphylococci, and Bacillus species [The intestinal microbiota also has the significant proteolytic capacity, converting ingested food protein and endogenous protein from host enzymes and mucin to shorter peptides, amino acids and their derivatives, short and branched-chain fatty acids, and gases, including CH and H2S ,53. Bact species ,53.12. These vitamins are important for both bacterial metabolism and the host [Colon bacteria are also able to synthesize some vitamins, especially B vitamins, including biotin, cobalamin, folate, nicotinic acid, pantothenic acid, pyridoxine, riboflavin, and thiamine, and vitamin K ,58. One the host .Indigestible polysaccharides, including cellulose and other substances, are fermented in the gut by anaerobic bacteria to obtain energy for microbial growth and production of short-chain fatty acids (SCFAs) . The thrClostridia sp. from the Firmicutes phylum strain [Butyrate is important representative of SCFAs and plays a key role in maintaining intestinal epithelial homeostasis as a preferred source of energy for colonocytes and their growth stimulator . Previoum strain . It is am strain .β, IL-10, IL-18, and by inducing the differentiation of naive T cells into regulatory T cells. A mild anti-inflammatory effect was also demonstrated for acetate and propionate. These two molecules have the ability to suppress the production of proinflammatory cytokines by stimulating TLR 4 [Butyrate also has anti-inflammatory effects by stimulating antigen-presenting cells to produce the cytokines TGF-ng TLR 4 .Butyrate metabolism is one of the current theories for the etiology of UC . It is tPropionate, like butyrate, can induce T-cell differentiation . PropionAcetate is the most abundant SCFA and is an important cofactor and metabolite for the growth of many microorganisms ,70. In tBacterial fermentation provides numerous intermediates, including fumarate, succinate, and lactate, but are commonly detected at low levels in the feces of healthy individuals due to their extensive use by other bacteria. For example, lactate is usually converted to propionate or butyrate by other bacteria and is therefore present at negligible levels in adult feces. However, in patients with UC, lactate can be detected in significantly higher amounts and could be a potential indicator of the disease ,72. It hLactobacilli and Bifidobacteria. It is therefore theoretically possible that probiotics or prebiotics can reduce the occurrence of gas in the intestines and also contribute to the suppression of some health problems [The gas is a product of microbial fermentation in anaerobic ecosystems, including the digestive tract. However, some anaerobically growing species produce no gas ,73. Thisproblems .4, NH3, H2, CO2, and H2S, among others. The key harmful and potentially toxic components are sulfides, which act as precursors to other sulfur-based components [Most microbially generated gas includes CHmponents . Howevermponents .2 composition of the flatus is up to 40% and appears to be exclusively of microbial origin [2 is produced by various intestinal microorganisms. The main producers are Bacteroides and Clostridium. There are three main microbial pathways by which H2 can be removed. These include sulfate dissimilation reduction, methanogenesis, and acetogenesis [The Hl origin . H2 is pogenesis .2 [Desulfovibrio. Sulfate can be ingested in the diet or released after microbial metabolism of sulfate mucins. These are glycoproteins that line the gastrointestinal tract and act as a protective barrier between the mucosal surface and the luminal contents [The dissimilative reduction of sulfates is provided by SRB. These microorganisms use sulfate as an electron acceptor to dissimilation organic compounds and H2 . The moscontents .2 to reduce sulfate to sulfide has effects on overall gas production in the colon by reducing the amount of free H2, thereby helping to prevent excessive gas accumulation. However, the highly toxic nature of the H2S produced can have pathological consequences for the host [The use of Hthe host . The SRB2 removal in the large intestine that also reduces the overall gas accumulation. Thus, methanogenic archaea and SRB compete for H2 in the gut, and the process that dominates depends on the amount of sulfate available. When sufficient sulfate is available, SRB are more easily recovered by H2 due to their higher affinity for the substrate [Methanogenesis is another mechanism of Hubstrate ,53.Meth2. The reason is the conversion of carbon dioxide (CO2) and H2 into acetate without releasing gas. However, this reaction is less energetically favorable than reduction by dissolving sulfate or methanogenesis [For host health, acetogenesis is probably the most preferred way to recycle Hogenesis .Acetoge2 is another quantitatively significant gas. CO2 can represent between 5% and 50% of the total volume of the flatus and is recycled by methanogenesis and to a lesser extent by acetogenesis. Unlike H2 and methane, CO2 can be generated by a number of processes, not just bacterial metabolism [COtabolism .2 recycling by disulfilization reduction of sulfate generates H2S, which is a cellular signaling molecule, but also a highly toxic substance for colon cells, and its production and amount is a cofactor of inflammatory bowel disease. The presence of methane in the colon has been linked to colorectal cancer, although this association may be due to disease rather than causation, as patients with these difficulties also suffer from slower intestinal passage through the colon, which benefits the growth of intestinal methanogens due to their slowly growing nature [The absence of SRB, methanogens, and acetogens would cause the individual to produce 5–10 times more gas than usual. Hg nature .2S by a process called sulfate dissimilation reduction and participate in the process of the biogeochemical cycle of sulfur in nature [2, which may be the only carbon source for autotrophic growth [Sulfate-reducing bacteria (SRB) are common in anaerobic areas of soil, wetlands, fresh and seawater, but also the colon of humans and animals or the oral cavity. These microorganisms dissimilate sulfate to Hn nature ,80. Thesn nature ,82,83,84c growth .2S produced by the reduction of sulfates is highly toxic to all living organisms, so it must be removed from the body, which is ensured by various mechanisms, including oxidation by microorganisms [2S is toxic not only to the host but also to its bacterial producers themselves. High concentrations (above 6 mmol) of H2S lead to the cessation of SRB growth and the development of inflammatory damage to the intestinal epithelium [Hrganisms ,86. It sithelium ,86.2S produced by SRB. It has been reported that the western diet contains more than 16.6 mmol of sulfates per day and the feces of approximately half of healthy individuals contain SRB, with 92% of the genus Desulfovibrio predominating [Desulfotomaculum, Desulfobulbus, Desulfomicrobium, Desulfomonas, and Desulfovibrio. The dominant species of SRB in the human intestinal tract is Desulfovibrio sp. [Bacteroides, Pseudomonas, Clostridium, and Escherichia [A diet rich in sulfate ions, such as foods preserved with sulfur oxides, causes an increase in the concentration of Hminating . Intestibrio sp. ,25,87,88brio sp. . Previoubrio sp. ,85. SRB herichia .2 and compete for this substrate with methanogenic archaea [Desulfovibrio represent 67–91% of the total number of SRB. Significantly fewer bacteria occur in the genera Desulfobacter (9–16%), Desulfobulbus (5–8%), and Desulfotomaculum (2%) [SRB grow on the surface of the mucosa or in the intestinal lumen. Living bacteria on the surface of the colonic mucosa, they interact with cells of the immune and neuroendocrine systems more closely than microorganisms in the intestinal lumen . The spe archaea . The prelum (2%) . ExampleDesulfovibrionales are straight or curved, motile rods. In relation to molecular oxygen (O2), they are strictly anaerobic. These bacteria are mesophilic or slightly thermophilic. They grow under neutral to slightly alkaline conditions. Sulfide, sulfide, and thiosulphate are reduced to H2S [Bacteria of the order d to H2S .Desulfovibrionaceae family oxidize organic matter to acetate. In addition to the gastrointestinal tract and feces, they can also be found in the genitals, oral cavity, and in the aquatic environment. Representatives of the genus Desulfovibrio are G−, curved, or spiral cells, mostly motile. The electron donor is usually H2, lactate, ethanol. The optimal temperature for growth is between 25 and 40 °C [Bacteria from the nd 40 °C .Desulfomicrobium includes ovoid, rod-shaped G− cells, motile or immobile. They oxidize H2, lactate, and malate. Organic substrates are incompletely oxidized to acetate. By type, representatives are difficult to distinguish. In addition to settlements in the intestine, they also occur in O2-free freshwater, in marine sediments, or water from oil fields [The genus l fields ,91.Desulfobacterales is diverse. They can form cocci or oval to regular rods, which can be curved. Sometimes we find them in clusters or threads. They are often mobile. Strictly anaerobic. This order also reduces sulfate, sulfide, and thiosine to H2S [The morphology of bacteria of the order e to H2S .Desulfobacter are G−. The cells are oval or rod-shaped, slightly curved, movable, and immobile. The donor of electrons is most often acetate, but it can also be ethanol, H2, and lactate [These bacteria are capable of complete oxidation of organic substrates. Representatives of the genus  lactate ,91.Desulfobulbaceae family includes cocci and rods, which are often motile. Simple organic substances serve as an electron donor and at the same time as a carbon source. They incompletely oxidize substrates and the final product is usually acetate. Mesophilic and psychrophilic organisms belong to the family Desulfobulbaceae. Representatives of the genus Desulfobulbus are G− type, mostly in the shape of an ovoid rod. The electron donor is propionate, but they also oxidize H2, lactate, ethanol [The  ethanol .Clostridium and related genera. Anaerobic G+ bacteria with low G+C content in DNA [Includes genus t in DNA .Peptococcaceae family includes cocci and sticks. We can also find saprophytic or clinically important species here. Representatives of the genus Desulfotomaculum are straight or curved rods, movable, and occurring individually. These bacteria form round to oval endospores, terminally or subterminally deposited, swollen cells. They are the only intestinal SRB that form spores. They grow at a temperature of 20–70 °C [The 20–70 °C .2S, which increases mucosal permeability and block butyrate metabolism. Increased mucosal permeability can lead to the activation of pathogenic T cell-mediated and innate immune responses through exposure to bacterial TLR ligands and antigen [Decreased concentrations of protective bacteria that produce short-chain fatty acids (SCFAs), such as butyrate, may increase mucosal permeability. Conversely, elevated concentrations of some bacteria may increase the production of toxic metabolites, such as H antigen .In the large intestine a specialized group of predominantly gram-negative anaerobes is present, known as SRB, which are capable of a process called the reduction of dissolving sulfates, reducing sulfates to sulfides. Sulfide is toxic to the intestinal epithelium. Thus, sulfate in the diet can selectively stimulate the growth of one group of bacteria with potentially harmful effects on the epithelium .2S, toxic to the intestinal mucosa by competition with SCFAs, which appears to be associated with an excess of SRB  in fecal samples [2S accumulation in human feces may predict the course of inflammation in the gut [2S toxicity is presented in Patients with active UC overproduce H samples . The est the gut . New opp the gut . The mec2 is the only electron donor for intestinal methanogenic archaea such as M. smithii. Therefore, there is competition for H2 between SRB and methanogenic organisms. When sulfates are present in sufficient amounts, SRB inhibit the use of H2 by methanogens in the dissimilation of the sulfate reduction process. Thus, the amount of sulfate in the diet may affect the competition of the substrate between SRB and methanogensin the large intestine. In the absence of sulfate in the diet, SRB were not found. Thus, the intensity of methanogenesis can be regulated by the introduction of sulfate, even when the SRB are low in the gut [H the gut ,10. The  the gut .Meta-analysis is a statistical process summarizing comparable data from a large number of scientific papers ,20,21. TDue to the exponential growth of scientific studies, it is becoming increasingly difficult to identify relevant information. It happens that the primary studies bring different conclusions, although they address the same research question . Meta-anProper meta-analysis is characterized by a thorough and disciplined search of the literature and, last but not least, discussions with experts in the field. A clear definition of the hypotheses to be investigated is important . For stuhttp://www.ncbi.nlm.nih.gov/pubmed), “ScienceDirect” (www.sciencedirect.com), “Institute of Scientific Information Web of Knowledge” (http://www.isiwebofknowledge.com) and “Google Scholar” (http://scholar.google.com) [The “Preferred Reporting Items for Systematic Reviews and Meta-Analyzes” (PRISMA) statement recommends a thorough search of key sources from at least one major database . There sgle.com) . Complexgle.com) .The meta-analysis presents knowledge about the size of the tested file, which makes it possible to identify connections between individual information and draw new conclusions. In the meta-analysis example, the results of each study are graphically presented in a forest plot, a method that uses an odds ratio (OR) and a confidence interval (CI) metric of 95% . Forest There are many ways to perform a meta-analysis. Commonly used statistical models for combining data in the meta-analysis are the fixed-effect model and the random-effect model. The fixed-effect model assumes that there is a common effect in all included studies and that the observed differences in results across the studies reflect random changes. The random-effects model assumes that there is no specific effect on all studies. The fixed-effect model only considers heterogeneity within the study, while the random effect model takes into account heterogeneity within and between studies .Meta-analysis is usually used for topics that do not have definitive studies or do not completely agree with the conclusions of these studies. Studies with little or no statistical power to confirm a fact often appear. By combining the results of several studies, sufficient statistical power can be achieved, and thus new facts can be discovered  . UC is a−1 can be found in dried fruits and vegetables, in nuts , cruciferous vegetables , in white and dark bread, soy flour and sausages [CD can affect any part of the gastrointestinal tract. UC is limited to the large intestine and rectum . This masausages .A comprehensive overview of the properties of intestinal SRB and their relationship to IBD was reported in the study published in 2020: “Hydrogen sulfide toxicity in the gut environment: meta-analysis of sulfate-reducing and lactic acid bacteria in inflammatory processes”. This study also highlights the positive effect of probiotics. A meta-analysis was used for statistical comparison of the respective studies. The meta-analysis included 16 studies, which were selected from the Web of Science database. RevMan software was used to create the meta-analysis. The meta-analysis shown in p < 0.05) difference between the incidence of SRB in healthy people and patients with advanced IBD. However, diamond, which summarizes all studies, suggests the occurrence of SRB mainly in people with IBD [All studies except one found a significant (with IBD . The trep < 0.05) difference was noted. All studies except the study of Rembacken et al. (1999) [The studies were conducted in such a way that the group was always divided into a control group and a probiotic group. Comparative studies show that most of them touch the zero-effect line and therefore have no significant benefit, except for the study by Ishikawa et al. (2003) , Miele e. (1999)  show tha. (1999) . A lowerα) is currently one of the most important drug target molecules used in several autoimmune diseases. Infliximab became the first TNF-α-inhibiting drug to be introduced into clinical practice. At the same time, it was the first registered biotechnologically prepared drug with the character of a monoclonal antibody. It is used in patients with IBD, but also a number of other autoimmune diseases . It can reduce the severity of the symptoms of the disease and cause remission [Tumor necrosis factor-alpha , but no heterogeneity was found in the colectomy rate after 12 months of treatment. Therefore, the method of random effects was used for the colectomy rate after 3 months, and the model of fixated effects for the colectomy rate after 12 months. A meta-analysis showed that the colectomy rate in the infliximab group was significantly lower than in the control group after 3 and 12 months of treatment, as shown in A systematic review from 2019 “In infliximab clinically treating ulcerative colitis: a systematic review and meta-analysis” is an evaluation of current UC treatment. This meta-analysis was performed in accordance with PRISMA criteria. All statistical analyzes were performed using RevMan 5.3 software. For a meta-analysis, studies comparing the efficacy of infliximab versus placebo, steroids, or immunosuppressants with UC were included. Only studies with clear information on the number of patients who were included and grouped for each therapy . The ratThis meta-analysis showed that infliximab reduced the rate of colon colectomy in patients with moderate to severe UC at 3 and 12 months. Additional meta-analyses were performed in the study to confirm that infliximab administration promotes the onset of remission compared to traditional drugs or placebo .Clostridium difficile  is an obligately anaerobic G+ sporulating rods [Clostridium difficile appears to be an important pathogen in patients with IBD and is associated with more severe disease development and increased mortality [ing rods . It is oortality .Clostridium difficile on surgical rate among ulcerative colitis patients: a systematic review and meta-analysis”, the authors evaluated the association between Clostridium difficile infection and the rate of colectomy in patients with UC. Statistical analysis was performed using RevMan X6 software (from Cochrane Collaboration) [In a 2015 study “The impact of oration) .Clostridium difficile were found to have statistically significantly higher risks of surgery compared to patients with UC without infection. There was no statistically significant heterogeneity among the included studies (p = 0.09). However, the number of studies included was small, so the result should be interpreted with caution [Patients with UC co-infected with  caution .UC is a chronic disease of the gastrointestinal tract with an as yet unclear etiology. This disease is manifested by an uncontrollable inflammatory immune response. In Europe, around 2.2 million people suffer from UC and the number of patients is still growing. To date, there is no effective treatment to ensure complete recovery of patients. Pharmacotherapy and other procedures only lead to the suppression of symptoms and increase the quality of life of patients, and if this therapy fails, a colectomy may be necessary. Butyrate plays a key role in maintaining intestinal epithelial homeostasis as a preferred source of energy for colonocytes. Butyrate metabolism is one of the current theories for the etiology of UC. It is thought that a lack of energy for colonocytes could lead to the onset of this disease. Butyrate has also been used experimentally in the treatment of colitis with numerous benefits for patients.2S by a process called sulfate dissimilation reduction. Intestinal SRB include the genera Desulfotomaculum, Desulfobulbus, Desulfomicrobium, Desulfomonas, and Desulfovibrio. SRB are not thought to be pathogenic in humans and animals, but increased production of toxic H2S increases mucosal permeability and blocks butyrate metabolism. UC is a complex topic, connecting a wide range of scientific areas.SRB are anaerobic bacteria that dissimilate sulfate to HClostridium difficile appears to be an important pathogen, which has been associated with more severe disease development and increased mortality in patients with IBD. Patients with UC who were co-infected with Clostridium difficile were found to have statistically significantly higher risks of surgery compared to patients with UC without infection. Idiopathic intestinal inflammation and assessing the effectiveness of their treatment are extremely urgent topics in biology and medicine. The results of the primary studies are inconsistent, so further analyzes are needed. The meta-analysis appears to be a suitable method for reaching valid conclusions in this area.Using meta-analysis, we compared data from many scientific papers. Results included an estimate of the effect of treatment or disease risk factors than individual studies. Since 2007, this statistical method has become increasingly popular among scientists. For this research, studies were found that use this method and solve the issue of UC. A meta-analysis from 2020 revealed a lower incidence of IBD within the group of subjects receiving probiotics and at the same time a higher prevalence of IBD in patients with a higher incidence of SRB. Another study evaluated the effectiveness of infliximab treatment. A meta-analysis showed that infliximab reduced the rate of colon colectomy in patients with moderate to severe UC.


Molar incisor hypomineralization (MIH) is a growing health problem, and its treatment is a challenge. The purpose of the present study was to evaluate and compare the perceptions, knowledge, and clinical experiences of MIH in general dental practitioners (GDPs) and paediatric dentists (PDs) in Spain.All dentists belonging to the College of Dentists of the Region of Murcia, in the South-East of Spain, were invited to participate in a cross-sectional survey. They were asked to complete a two-part questionnaire including sociodemographic profiles and knowledge, experience, and perceptions of MIH. Data were analysed using Pearson’s chi-square test, Fisher’s exact test and Cramer’s V test.p <  0.001). Only 23.80% of GDPs claimed to have made a training course on MIH. With respect to the aetiology, chronic medical conditions (p = 0.029) and environmental pollutants (p = 0.008) were the only factors that showed significant between-group differences. Durability (p = 0.009) and remineralization potential (p = 0.018) were the factors where there was a between-group difference in the choice of the restoration material. In the case of post-eruptive fractures and opacities, the preferred material for both groups was resin-modified glass ionomer (RMGIC). However, in incisor lesions, composite was the material of choice for both groups, with significant differences (p = 0.032) in the use of glass ionomer. Most respondents expressed a need for continuing education on MIH.The overall response rate was 18.6% (214/1147). Most respondents were aged 31–40 years (44.86%), with more than 15 years of professional experience (39.72%). They worked mainly in the private sector (84.58%) and were licensed in dentistry (74.30%): 95.45% of PDs had detected an increase in the incidence of MIH in recent years (Spanish dentists perceived an increase in the incidence of MIH. The material of choice was RMGIC for non-aesthetic sectors and composite for incisors. Dentists believe it is difficult or very difficult to manage MIH, since the long-term success of restorations of MIH lesions is compromised because resin adhesion is not good. Both GDPs and PDs believe they need more training on the aetiology, diagnosis, and treatment of MIH. The term molar incisor hypomineralization (MIH) was described by Weerheijm, et al. in 2001 and adopted by the international dental community due to a consensus at the Congress of the European Academy of Paediatric Dentistry in Athens in 2003 [Historically, in the dental literature, a wide variety of terminology and definitions for enamel defects in hypomineralized molars, with or without post-eruptive enamel fractures have been used: enamel opacity not caused by fluoride, internal enamel hypoplasia, non-endemic enamel speckling, opaque stains, idiopathic enamel opacities, and enamel opacity. Some terms simply describe the pathology, while others bear the name of the causal agent . DespiteClinically, the form of presentation and the severity of MIH-affected teeth may be asymmetrical in the same patient and vary from mild opacities to severe post-eruptive breakdown that may affect from one to four first permanent molars . MIH mayThe global recorded prevalence of MIH ranges from 2.4 to 40% and differs between countries , 8. TherThe high prevalence and incidence of MIH, the poor quality of life of paediatric patients and the difficult clinical management, have led to numerous studies on the perception, diagnosis and management of MIH through surveys of dentists, both general dental practitioners (GDPs) and paediatric dentists (PDs) –22. In Sn = 1147) in March 2019. The email explained the study, stated that participation was anonymous and voluntary, and enclosed a link to accede to the survey without signing in to Google. The study researchers had no access to the personal data of participants. Participants were asked to complete the online questionnaire on their own time. A reminder email was sent 2 weeks after initial distribution. The survey was online for 1 month.The study was approved by the Bioethics Committee of Murcia University (Reference Number: 2255/2019). Google Survey software was used to develop the survey that was subsequently emailed to all members of College of Dentists of Murcia , educational background (main area of work), perception (changes in the incidence of MIH lesions in recent years), clinical appearance , prevalence , participants’ attitudes, knowledge (aetiology) and practice in MIH management and, finally, restorative options in MIH management .In the second part of the questionnaire, two clinical situations with illustrative photographs and a written case description were suggested to dentists. In clinical case 1 Fig. , dentistThe questions in the first section of the questionnaire, where it says “tick one option”, and the first clinical case have only one answer. The second clinical case, and the questions where it says “choose the corresponding answers”, are multiple choice, so the number of responses could be greater than the number of respondents.p-value < 0.05 and significance level 0.05). Cramer’s test was used to determine whether the relationship was strong or weak.Study data were processed and analysed using the R statistical package. A simple frequency distribution was made. Independent variables (sociodemographic variables: Table n = 214): 69.16% were GDPs or other specialties (n = 148), and 30.84% were PDs (n = 66).Of the 1147 dentists invited to participate, 216 responded. Two surveys were eliminated because they were not completed correctly, resulting in a response rate of 18.66% .In terms of prevalence, 59.45% of GDPs found that < 10% of their patients had MIH, while 50.00% of PDs said that 10–25% of their patients had MIH. In addition, 95.45% of PDs had detected an increase in the incidence of MIH in recent years  and GDPs (58.11%) and were most often diagnosed in the permanent teething.p = 0.029) and environmental pollutants (p = 0.008) were the only factors that showed significant differences between the two groups.The knowledge of respondents regarding the aetiology of MIH is shown in Table p < 0.001). PDs stated that the information they obtain on MIH basically comes from face-to-face continuing education (44.26%), while the Internet was the source of choice for GDPs (36.15%) (p = 0.005), with widespread demand for information on the aetiology, diagnosis and treatment of MIH.As for the difficulty of managing MIH, the most frequent response was that it is considered a challenge and that GPDs (76.19%) had not received any information on this (p = 0.009) and remineralization potential (p = 0.018). As for the material of choice in cases of post-eruptive fractures, RMGIC was the most widely used by both groups. However, there were significant differences in the use of glass ionomer cement (GIC) (p = 0.048) between GDPs (12.21%) and PDs (18.40%). No significant differences were found in the materials used to restore opacity, with RMGIC again being the first choice in both groups. However, in the case of the treatment of lesions in the incisors, composite was the material of choice in both groups, with significant differences (p = 0.032) in the use of RMGIC between GDPs and PDs.The results on restorative treatments in MIH are shown in Table In the second part of the questionnaire, in the first clinical case Fig. , the matDespite the high prevalence and increased incidence of MIH in Spanish paediatric patients, this is the first study to provide information on the perception and knowledge of the aetiology and diagnosis of MIH and patient management strategies of Spanish GDPs and PDs.We used an online survey to avoid the low response rate obtained in postal surveys . The resIn Spain, dentistry is mostly private, which is reflected by the responders (84.58% private), unlike countries such as Norway  or AustrWe found that PDs had twice the perception of patients with MIH lesions compared with GDPs, a situation reflected in countries such as Iraq , MalaysiThe most recognized MIH lesion in both study groups was yellow/brown lesions, as it was in other countries , 15–18. In 2012, hypomineralization of the primary teeth was described, mainly in the second primary molars (HSPM). This is known to be associated with an increased risk of hypomineralization in the permanent molars, although the absence of HSPM does not exclude future MIH , Kuwait In general, and as in other studies –18, 21, A significant percentage of both GDPs and PDs responded that they found the management of MIH “somewhat difficult”. This is because these patients have increased anxiety  and tootAchieving correct restoration and long-term success is what worries dentists the most (48.84%). It is known that etching with orthophosphoric acid creates faulty etched patterns , that reThere are many reported treatment options for the restoration of teeth with MIH lesions: fluoride and/or CPP-ACP remineralization systems, silver diamine fluoride, pit and fissure sealants, resin infiltrations, conventional and modified glass ionomers with resin, resin composite, amalgam, preformed crowns, and even extractions, always depending on the severity of the lesion .The potential for remineralization of material in restoration is the most relevant factor in the choice of materials (25.3%), significantly worrying GDPs more than PDs. In fact, the most commonly used material to restore post-eruptive fractures is RMGIC, followed by composite and both are used equally by GDPs and PDs. GIC is the third material of choice and is used proportionally more by PDs than GDPs. This may be because they treat younger children and use it as filling material in atraumatic restorative treatments or for interim restorations. Durability, which is one of the most relevant factors in material choice, is therefore significantly less decisive for PDs than for GDPs.There are studies that show GIC 81%) was used more than RMGIC (44.3%), which is justified by the greater fluoride release . However1% was usWith respect to enamel opacity, the materials chosen were the same in both groups; first RMGIC followed by composite. However, in incisor lesions, composite was the material of choice, due to aesthetic concerns, followed by RMGIC and resin infiltrations. PDs use significantly more RMGIC to restore incisor enamel lesions.The adhesion, durability and potential for remineralization were also decisive in the choice of material by most dentists from other countries , 16, 18.In clinical case 1 (Fig. In clinical case 2, the preferred option for GDPs was to remove the tissue seemingly most affected and restore with glass ionomer, compared with PDs whose option was not to remove any dental tissue and use glass ionomer to restore. This shows a trend towards less invasive treatment by PDs, as described by other reports .The study had some limitations. First, the response rate was low, although this is characteristic of online surveys. Secondly, it was aimed at dentists from one Spanish region. Therefore, extrapolation of the results to the rest of Spain, should be made with caution, as sociodemographic characteristics may vary. However, the survey may serve as a starting point for the introduction of Spanish guidelines or protocols on the correct care of children with MIH.Spanish dentists perceive that the incidence of MIH has increased in recent years. They believe it is difficult or very difficult to manage MIH, since the long-term success of restorations of MIH lesions is compromised because resin adhesion is not good. They use RMGICs more frequently, taking advantage of their remineralizing potential, except in the incisors, where they use composites. Both GDPs and PDs think they need more training on the aetiology, diagnosis, and treatment of MIH. The introduction of national guidelines that serve as a reference manual for all continuing education courses would improve the management of MIH.Additional file 1.


Aging causes some unfavorable morphological and functional changes, such as the decline in bone mineral density (BMD) and physical function. Moderate-to-vigorous physical activity (MVPA) and sedentary time seem to be related with these alterations, but the impact of distinct patterns remains unclear. The aim of this study was to cross-sectionally and prospectively assess the association between objectively measured MVPA and sedentary patterns (bouts and breaks) with BMD and physical function in older adults. The study considered 151 Brazilians (aged ≥ 60 years), out of which 68 participants completed 2-year follow-up measurements. MVPA and sedentary patterns were measured by means of accelerometry, BMD—) by means of dual-energy X-ray absorptiometry (DXA), and physical function—by means of physical tests. In older women, sedentary bouts >60 min were inversely associated with handgrip strength . The prospective analyses showed that changes in sedentary bouts (20 to 30 min and >60 min) were inversely associated with changes in the lumbar spine’s BMD  and the lumbar spine’s T-score , respectively. In older women, sedentary patterns are cross-sectionally associated with handgrip strength and prospectively associated with BMD independent of MVPA. The natural process of aging causes some unfavorable morphological and functional changes, such as the decline in bone mineral density (BMD)  and physBehavior patterns such as moderate-to-vigorous physical activity (MVPA)  and sedeOlder adults are the most sedentary group in the population, spending approximately 65 to 80% (9 to 10 h/day) of their waking hours engaged in sedentary pursuits ,13,14 wiCurrently, there is a paucity of research assessing the associations of sedentary patterns (breaks and bouts) with BMD and physical function in older adults ,11, espeThe present study is a cohort study performed between January 2015 and February 2017 in the city of Presidente Prudente, São Paulo State, Brazil. The main goal was to analyze the influence of physical activity on sarcopenia, sarcopenic obesity, and functional disability in older adults. In this paper, we chose to present cross-sectional and prospective data.Sample selection was performed by convenience sampling. The inclusion criterion was as follows: older adults aged 60 years or older living in Presidente Prudente who attended all the evaluations at the laboratory. The exclusion criteria consisted of the following factors: living at a long-term institution; presence of a disease that could lead to muscle mass reduction, such as cancer, HIV/AIDS, tuberculosis, and chronic kidney disease. A minimum sample size of 99 participants was identified based on the expected correlation coefficient of 0.287 between appendicular lean mass and physical activity  using thData collection was performed at the Center for Studies and Laboratory of Assessment and Motor Activity Prescription (CELAPAM), Department of Physical Education, FCT/UNESP. The participants who accepted the invitation to participate in the project signed the informed consent forms. All the protocols were reviewed and approved by the Ethics Research Committee (Process CAEE 26058114.3.0000.5402).BMD of the proximal femur and lumbar spine was assessed using dual-energy X-ray absorptiometry (DXA) . All the examinations were performed according to the manufacturer’s recommendations. The presence of osteopenia or osteoporosis in the proximal femur or lumbar spine (L1-L4) was defined as T-score values ≤ −1.0 or ≤−2.5, respectively .Physical function was estimated using three functional tests: handgrip strength, gait speed, and timed up and go tests as described below.The handgrip strength was measured using a Camry digital dynamometer model EH101 . The test was performed with the participant sitting in a chair without arm support, having their shoulder slightly adducted and elbow of the dominant arm flexed at 90° with the forearm and wrist in the neutral position. The participants were instructed to press the dynamometer as strongly as possible twice at intervals of one minute between each attempt. The highest handgrip strength value (kg) was recorded . PreviouThe 4-m walk test was used to assess the participants’ gait speed. The participants were advised to walk in a natural way as if walking indoors. Any habitual apparatus for walking  could be used . The walThe timed up and go (TUG) test was performed to evaluate gait speed and dynamic balance (time in seconds). This test consisted of the participant sitting in a stable chair with arm support (hips and back completely leaning on the seat), getting up from the chair, walking three meters, turning around, returning, and sitting back in the chair. The individual could use the arms of the chair to move from the sitting position to the standing position and vice versa . For thiActigraph GT3X  accelerometer motion sensors were used to estimate daily MVPA and sedentary patterns. Older adults were instructed to wear accelerometers attached to the waist for five consecutive days during all waking hours of the day, except when sleeping and when performing water-based activities .For data analysis, the ActiLife6 software  was used. Only days with at least 600 min were considered valid. Periods of 60 consecutive minutes of zero counts were considered the non-wear time . To be iEach minute in which the accelerometer counts were below 100 was considered sedentary time. Total sedentary time was the sum of all sedentary minutes when the accelerometer was worn. A bout was considered a specific period of continuous sedentary time in which the accelerometer count was less than 100 counts/min with no interruption. Bouts of continuous sedentary time (counts/minute < 100) of 20 to 30 min, 30 to 60 min, and >60 min were used in the analyses.A break in sedentary time was defined as an interruption (lasting at least 1 min) in sedentary time in which the count was above 100 counts/min.The cut points for defining the intensity of physical activity were analyzed following the recommendation established by Troiano et al. . Low int® Antropométrica, São Paulo, Brazil) and a fixed stadiometer , respectively. Body mass index (weight (kg)/height2 (m)) was calculated.Body mass and stature were measured using an electronic scale , and income  of the study population was obtained through a self-reported interview.t-test. Associations with BMD and physical function were initially explored using the Pearson’s correlation coefficients. Covariates with the p-value < 0.05 were included in the linear regression models, which were used to assess the relationships of MVPA and sedentary patterns with BMD and physical function. The results are reported as unstandardized β-coefficients with 95% confidence intervals. Cross-sectional regression analyses were performed for the baseline data controlling for age, BMI, osteopenia and osteoporosis, and MVPA when sedentary patterns (bouts or breaks) were the exposure of interest or the total sedentary time when MVPA was the exposure of interest. For the prospective analyses, the changes for both exposures and outcomes were calculated (follow-up minus baseline) and linear regression models were adjusted for the baseline age, BMI, osteopenia and osteoporosis, and the aimed exposure and outcome, as well as MVPA (changes) when sedentary patterns were the exposure of interest or the total sedentary time (changes) when MVPA was the exposure of interest. Statistical analysis were performed using SPSS software  version 23.0 and the significance level was set at 5%.The prevalence of osteopenia and osteoporosis in both analyses  according to sex was analyzed using the chi-squared test. Descriptive statistics consisted of the mean and standard deviation for all the relevant variables. The means for each variable were compared between sexes using an independent The participants’ characteristics are shown in In cross-sectional analyses, 84.1% of the older men had a normal total proximal femur T-score, 13.6% had osteopenia, and 2.3% had osteoporosis. Among older women, 44.9% had a normal total proximal femur T-score, 49.5% had osteopenia, and 5.6% had osteoporosis.p ≤ 0.001).Regarding the lumbar spine T-score, 75.0%, 20.5%, and 4.5% of the older men had a normal lumbar spine T-score, osteopenia, and osteoporosis, respectively. Among older women, the observed values were 38.3%, 46.7% and 15% for a normal lumbar spine T-score, osteopenia, and osteoporosis, respectively, with older women showing higher prevalence of osteopenia and osteoporosis in both regions .Regarding the lumbar spine T-score, 78.9%, 21.1%, and 0% of the older men had a normal lumbar spine T-score, osteopenia, and osteoporosis, respectively. In older women, the observed values were 26.5%, 57.2%, and 16.3% for a normal lumbar spine T-score, osteopenia, and osteoporosis, respectively, with older women showing higher prevalence of osteopenia and osteoporosis in both regions  may have beneficial effects on spine BMD.It is known that prolonged sitting is detrimental to bone health due to the absence of the contractile action of the skeletal muscle on bone as well as the absence of gravitational load on the skeleton . AccordiAlthough sedentary time has been negatively associated with bone health, the exact amount of time in which one has to spend in a sedentary behavior for deleterious effects to occur on BMD remains uncertain . In the On the other hand, there is still no consensus regarding the relationship between BMD and sedentary time in older men. Hind et al.  found thBeyond sedentary bouts, the number of times people break the sedentary time up is an important sedentary pattern to be considered. Braun et al.  found a Muscle strength is one of the most important components of physical fitness for performing activities of daily living (ADL) and maintaining a functional capacity in older adults. Reducing the time in activities that stimulate muscle contraction can affect the amount of muscle mass and, consequently, muscle quality ,17,33. FIn the appendicular skeleton, the trabecular bone transfers mechanical loads from the articular surface to the cortical bone, whereas in vertebral bodies, it represents the main load-bearing structure. Mechanical properties of the bone tissue and architecture of the trabecular bone are two main factors that determine the mechanical properties of the trabecular bone. Fragility fractures that arise in the context of metabolic bone diseases such as osteoporosis usually occur in regions of the trabecular bone .In cross-sectional analyses within this study, negative associations were found for sedentary bouts >60 min with handgrip strength in older women. However, in our prospective analysis, we found no associations between sedentary bouts and muscle strength, which is similar to the result reported by Keevil et al.  in a cohRegarding the breaks in sedentary time, we did not observe a significant relationship for the number of breaks in sedentary time with physical function in older adults. These results differ from those previously found by Sardinha et al.  and van Some limitations to be mentioned in the present study are the fact that some parameters that may be related to BMD have not been investigated, such as education level, calcium, vitamin D, nutrition, medication, alcohol, and bone fractures. In addition, a reduced sample and the fact that accelerometers are not sensitive to detect all activities, such as biking, standing, and upper body movement, may also be considered relevant limitations. Accelerometers are not able to identify the moment of transition between sitting and standing . NeverthA major strength of this study is its novelty in the sense that we investigated beyond the simple associations of the total sedentary time with BMD and physical function in older adults and looked into the relationship of sedentary patterns (breaks and bouts) with these outcomes both from a cross-sectional perspective and considering a 24-month longitudinal perspective. In addition, despite the aforementioned limitations in the use of accelerometry to assess sedentary patterns and MVPA, it is a widely used objective method within the physical activity research field, thus allowing comparison with previous studies.In older women, sedentary patterns are cross-sectionally associated with handgrip strength and prospectively associated with BMD independent of MVPA. The findings from the present study indicate that, in the long term, limiting continuous prolonged sedentary bouts, especially longer than 60 min, may have beneficial effects on BMD. Thus, in addition to some preventive strategies such as engagement in physical activity, intervention programs must aim to reduce sedentary time in the daily routine of this specific population, as it can promote benefits to BMD.


Java Island is one of the islands in Indonesia which has local sheep breeds with specific characteristics and native development geography in certain regions. This study aimed to determine the genetic profiles and maternal origin of six local sheep breeds on Java Island.This study was conducted by identifying the profiles of complete mitochondrial DNA (mtDNA) displacement loop (D-loop) region sequences on a total of 22 individual in six local sheep breeds on Java Island, including Javanese thin-tailed (JTT), Javanese Fat-Tailed (JFT), Batur (BTR), Wonosobo (WSB), Garut (GRT), and Priangan (PRG) sheep. The D-loop region was amplified using specific primers, and the polymerase chain reaction (PCR) was performed. The PCR products were purified and sequenced.The mtDNA D-loop analysis identified 21 haplotypes in the analyzed 22 animals with 123 polymorphic sites (V) consisting of 60 singleton variable sites (S) and 63 parsimony informative sites (P). Within all breeds tested, the haplotype diversity, the average number of pairwise differences (K), and nucleotide diversity (Pi) were 0.99567, 25.36364, and 0.02153, respectively. The genetic distance (D) within groups and between groups was 0.001-0.006 and 0.004-0.036, respectively. The phylogeny resulted in the presence of two haplogroups (Hap), which are 5 Hap A and 16 Hap B. All JTT, JFT, BTR, and WSB breeds were in the same cluster in Hap B, whereas GRT and PRG breeds were in clusters in both Hap A and Hap B.The high genetic diversity in six local sheep breeds on Java Island suggests that they originated from different genetic sources. JTT sheep have closer genetic relationships to JFT, BTR, and WSB sheep, and they are close to European sheep, whereas GRT sheep have closer genetic relationships to PRG sheep. Both are closer to Asian sheep than to European sheep. Indonesia is an archipelago country that has specific characteristics in each Island and a variety of natural and human resources. Java Island is one of the islands in Indonesia, which consists of four provinces  and two special regions (Jakarta and Yogyakarta). In general, these regions have similar characteristics, but in certain regions, they have unique characteristics regarding geographic features, natural resources, social, and cultural background. The character of the region and different natural resources dictate the type a socio-cultural community that will be established, at least with respect to farming activities. Farming activities adjust to the conditions of the area, including the selection of livestock that is suitable for rising. Sheep are one of the many livestock raised by farmers on Java Island that have a notable value in society ,2. SheepOne method to determine genetic diversity, relationships and origins of local Indonesian sheep are to examine the mitochondrial DNA (mtDNA) profile. The mtDNA is an important genetic study tool in population and systematic molecular genetics because it has many copies with a rapid evolutionary rate and is inherited . One parThis study aimed to determine the complete mtDNA D-loop sequences of six local sheep breeds on Java Island, Indonesia. The results of this study can be used to determine genetic diversity, genetic distance, and relationships between local sheep breeds and other species of sheep on Java Island. Furthermore, the results can be added to databases and used to complement previous studies on genetic diversity and relationships of local Indonesian sheep, especially on Java Island.This study was approved by the Animal Care and Use Committee of the Faculty of Veterinary Medicine, Universitas Gadjah Mada, with the ethical clearance number 002/EC-FKH/Int/2019, and the National Political and Unity of Yogyakarta Special District with the approval number 074/1850/Kesbangpol/2019.This study was conducted using blood samples from 22 individual sheep in six populations of local sheep breeds on Java Island and is presented in http://primer3.UT.ee//) [Ovis aries (GenBank accession number: AF010406.1). The mtDNA D-loop region primer sequences were Alek-DLF: 5’-GAAGAAGCTATAGCCCCACT-3’ and Alek-DLR: 5’-GATTCGAAGGGCGTTACT-3’ that generated 1397 bp of the PCR product. The PCR reaction consisted of 4 μL of DNA template, 25 μL of KAPA2G Fast Ready Mix+Dye (Kapa Biosystems Ltd.), 2 μL of forward primer, 2 μL of reverse primer, and 17 of ddH2O. The PCR amplification was conducted using Cleaver GTC96S (Cleaver Scientific Ltd.) according to the program: 6-min of pre-denaturation at 94°C, followed by 35 cycles, each consisting denaturation at 94°C for 30-s, primers annealing at 47°C for 40-s, extension at 72°C for 90-s, then ending with a final extension at 72°C for 5-min, and storage at 4°C. The PCR product was visualized using 1.5% agarose gel, and electrophoresis was run at 100 mV for 30-min. The result of amplification could be seen on the UV illuminator. The purified PCR products were sequenced by 1st BASE-Asia, Malaysia.The DNA was extracted based on the manufacturer’s standard protocol using PureLink™ Genomic DNA Mini Kits . The mtDNA D-loop was amplified directly from the genomic DNA by polymerase chain reaction (PCR). The primer was designed using the Primer3 online version 4.1.0 program (UT.ee//)  based onThe product length of the D-loop region sequences amplification for each individual is 1800 bp and was analyzed using the molecular evolutionary genetics analysis (MEGA) version 7.0 software  . The D-lReference mtDNA D-loop region sequences of wild and domestic sheep of known haplogroup (Hap) types  were dowOvis aries (AF010406.1), was equal to 1180 bp. However, differences were found in the sequence at 908 where C nucleotide insertion and deletion in the sequence at 1037 except in the GRT2 sample, which still has a T nucleotide at the site.All local sheep breeds on Java Island have D-loop sequences of 1180 bp except for one sample (GRT2) that has a length of 1181 bp. The length of this sequence, as the long sequence of the D-loop on The alignment of mtDNA D-loop nucleotide sequences of local sheep breeds on Java Island enabled polymorphic site data to be obtained and is presented in The parameters of genetic diversity and HD of the mtDNA D-loop region in six subpopulations of local sheep breeds on Java Island are as shown in The value of genetic distance within and between groups is presented in The phylogenetic tree of 21 haplotypes in six subpopulations of local sheep breeds on Java Island and mtDNA D-loop sequences of wild sheep and domestic sheep categorized by Hap from GenBank are presented in O. aries (AF010406.1), which is 1800 bp, but there is a difference at the 908 site where the C nucleotide is inserted. A deletion occurs at the 1037 site except for GTR2 samples . The nucleotide composition of six local sheep on Java in this study was almost similar to Tibetan sheep [The sequence length, in this study, is the same as the length of the D-loop sequence in  samples , which h samples , but stian sheep ,27, Smalan sheep , and Alpan sheep , with thGenetic diversity, besides reflecting differences in traits between individuals, also reflects small differences in nucleotides . Geneticet al. [Genetic distance is the estimate of the closeness of genetic relationships or phylogenetics between the population, according to the Nei . Geneticet al. , who fouet al. , the valet al. . Apart fet al. ,42, compet al. ,43, wholet al. ,45, and et al.  and/or tAn HD value in the range ≥0 <0.5 was included in the low category, whereas an HD value in the range >0.5 ≤1 was included in the high category ,35. BaseThe results of analyzing 22 individual samples were used as the basis to form 21 unique haplotypes and one shared haplotype. A comparison of the number of individual samples and the number of haplotypes  indicateet al. [Domestic sheep were divided into five Hap, namely, A, B, C, D, and E . Based oet al.  that Indet al. ,55. Thiset al. , easternet al. , and Kaiet al.  that foret al. , Romaniaet al. , and Ibeet al.  where Haet al. , Tibetanet al. , and natet al. , where HOvis musimon (European Mouflon)  . In cont , and from Europe, including Britain, Portugal, and the Netherlands/Dutch, traded in this region. Java Island is one of the islands in the Nusantara (Indonesia) that is a strategic destination of various regions for exploring and trading [Indonesia is an archipelago country between two continents  and two oceans (Indian and Pacific). In the past, this area was named  trading ,65. This trading . No studThis study shows that most local sheep breeds on Java Island are included in Hap B, which are sheep that have a close relationship with European sheep, even though this country is on the Asian continent. However, there are also local sheep breeds on Java Island that are included in Hap A. This indicates that the sheep that developed on Java Island were the result of crossing Asian and European sheep. However, this study is still limited to a small number of samples, but it can be used as a database for further research. Observations with data from mtDNA are used to determine the maternal inheritance line. It is necessary to have additional observations of the male’s line so that both maternal and paternal lines can be known to find out more about the origin of developing breeds. In Indonesia, aside from Java Island, there are still other local sheep breeds that have been established by the Ministry of Agriculture, whereas others have not despite their long development in the region. Thus, further research on these breeds still needs to be done. In-depth studies need to be performed on the genetic profiles of local Indonesian sheep breeds with more samples, a wider area, more comparative data, and various other methods that can describe in more detail the origins, characteristics, and productivity of Indonesian sheep breeds.The high genetic diversity of six local sheep breeds on Java Island suggests that they originated from different genetic resources. The JTT sheep have a closer genetic relationship to the JFT, BTR, and WSB sheep, and they are close to European sheep. In contrast, the GRT sheep have a closer genetic relationship to the PRG sheep. Both are closer to Asian sheep than they are to European sheep.WTA supervised the present study, and IGSB and RW designed and coordinated the study. AI performed the experiment, analyzed the data, and wrote the manuscript. The final manuscript has been read and developed in consultation with all authors. All authors read and approved the final manuscript.


Sardinella aurita) and the white steenbras (Lithognathus mormyrus) versus two Lessepsian species, the Randall’s threadfin bream  and the Lessepsian lizardfish (Saurida lessepsianus). A molecular method was used to detect NNV in all four fish species tested. In N. randalli, a relatively newly established invasive species in the Mediterranean Sea, the prevalence was significantly higher than in both indigenous species. In S. lessepsianus, prevalence varied considerably between years. While the factors that influence the effective establishment of invasive species are poorly understood, we suggest that the susceptibility of a given invasive fish species to locally acquired viral pathogens such as NVV may be important, in terms of both its successful establishment in its newly adopted environment and its role as a reservoir ‘host’ in the new area.Viruses are among the most abundant and diverse biological components in the marine environment. In finfish, viruses are key drivers of host diversity and population dynamics, and therefore, their effect on the marine environment is far-reaching. Viral encephalopathy and retinopathy (VER) is a disease caused by the marine nervous necrosis virus (NNV), which is recognized as one of the main infectious threats for marine aquaculture worldwide. For over 140 years, the Suez Canal has acted as a conduit for the invasion of Red Sea marine species into the Mediterranean Sea. In 2016–2017, we evaluated the prevalence of NNV in two indigenous Mediterranean species, the round sardinella ( Betanodavirus (Nodaviridae). NNV is globally distributed and has been detected in 177 susceptible marine species, while epizootic outbreaks have been reported in 62 of them [Marine organisms typically host diverse communities of viruses, bacteria, archaea, and fungi ,2,3. Mar of them . The dis of them ,6,7,8. T of them . Among a of them ,11. The  of them ,13, and  of them ,15,16,17 of them . Thus, l of them ,20. Betanodaviruses have been clustered into four genotypes, currently accepted by the International Committee on Taxonomy of Viruses (ICTV) as official species of this genus: striped jack nervous necrosis virus (SJNNV), tiger puffer nervous necrosis virus (TPNNV), barfin flounder nervous necrosis virus (BFNNV), and red-spotted grouper nervous necrosis virus (RGNNV) [The genome of NNV consists of two segments, RNA1 (3.1 Kb) and RNA2 (1.4 Kb), which code for the RNA-dependent RNA polymerase and the coat protein. Based on a phylogenetic analysis of the T4 variable region within the RNA2 segment,  (RGNNV) .Sardinella aurita) and the white steenbras (Lithognathus mormyrus), and two Lessepsian species, Randall’s bream  and lizardfish (Saurida lessepsianus). These four species were selected due to their ecological and economic importance as major species in the eastern Mediterranean fisheries. In addition, NNV detected in various species was genetically characterized.In our previous study , NNV wasL. mormyrus (n = 20), S. aurita (n = 28), N. randalli (n = 31), and S. lessepsianus (n = 30) were caught and brought on ice to the laboratory. All specimens were caught along the Israeli Mediterranean shoreline by bottom trawling during the annual national monitoring project conducted in November 2017 by the Israel Oceanographic and Limnological Research Institute (IOLR) and during the surveys conducted between October to December 2017 by the Fisheries and Aquaculture Department of the Israel Ministry of Agriculture. Weight and Total Length measurements were taken, and visual inspections were carried out. All specimens were aseptically dissected and brain tissue samples were collected according to fish necropsy protocol [In October–December 2017, a total of 109 specimens of four wild fish species: protocol . All samTM Reverse Transcription System  with random primers according to the manufacturer’s instructions.Total RNA from brain tissue was extracted by using the EZ-RNA total RNA isolation kit  according to the manufacturer’s instructions. The RNA concentration and quality were estimated using NanoDrop One (Thermo Scientific), and the extracted RNA was stored at −80 °C until use. Real-time Reverse Transcription PCR (RT-PCR) was performed to generate cDNA of total RNA using the GoScriptFor genotyping the NNV strains, all samples were subjected to PCR targeting both genomic segments using primers previously described by ,24 targehttp://tree.bio.ed.ac.uk/software/figtree/). PCR amplicons were purified by ExoSAP-IT (Affymetrix), followed by Sanger sequencing at the Macrogen Europe Laboratory . Sequences were aligned using the MEGA7 software  and compp-value < 0.05 was considered significant.The statistical significance of the prevalence of NNV between various fish species was determined by the Chi-Square test of independence using IBM SPSS Statistics for Windows, version 20.0 . For all tests, a p > 0.05) in total prevalence of NNV was found in the same sampled species when comparing the two consecutive years 2016 [S. lessepsianus, when comparing to the indigenous species, was significantly higher , whereas in the case of the two-year prevalence of NNV in N. randalli as compared to the indigenous species, the higher values were highly significant . Hence, the prevalence of NNV in the Lessepsian species was significantly higher than in the indigenous species in both consecutive years together . However, the main prevalence values for this finding were dominated by N. randalli, in which the prevalence of NNV was significantly higher than in the other three species in 2017 alone , as well as in the combined two-year data . The RNA1 and RNA2 phylogenetic analysis of NNV revealed that all samples were clustered with the RGNNV genotype  and no reassortants were found , it was revealed that the parasite assemblage of F. commersonii, after migration to the Mediterranean Sea, was reduced in its natural specialist parasites. At the same time, native generalist parasite species were acquired in the new environment, in agreement with the enemy release hypothesis [N. randalli was first reported in the Mediterranean Sea in 2005 [N. randalli, these are not yet understood. Some possible explanations for the establishment of invasive species have been suggested, for example, favorable temperature, foraging activities, and partitioning of biological niche [N. randalli, but a low ability to produce lethal disease due to different host immune mechanisms [N. randalli after invasion. Unfortunately, there are no data on prevalence levels of NNV in N. randalli in the Red Sea (nor in any Red Sea fish species). It is well known that the RGNNV genotype, which affects tropical and temperate fish species, is the most widely distributed NNV-genotype, and has a high recorded number of susceptible species [https://isramar.ocean.org.il/isramar_data/TimeSeries.aspx). In an experimental infection conducted on European seabass (Dicentrarchus labrax), it was shown that RGNNV would be more virulent at 25–30 °C than at 20 °C [D. labrax. Further studies should determine whether the high prevalence of this virus in the Lessepsian population is due to high resistance of the host, to temperature-related factors, or to other unknown factors that contributed to its successful establishment in the Mediterranean Sea in a relatively short time. We also suggest that further studies should include isolation of the viral agent in order to evaluate whole-genome sequencing (WGS) and provide a wider molecular insight of wild-NNV-isolates. Moreover, the histopathology of these wild fish species in future studies will enable us to better understand whether the histopathological manifestation in wild species differs from our knowledge of common mariculture species. This study warrants further investigations to try isolating the virus to determine whether it is viable, and to perform a histopathological analysis in order to verify whether it causes damage to the nervous tissues of these two Lessepsian species.The prevalence of NNV in 109 individual fish belonging to two indigenous and two Lessepsian species from the eastern Mediterranean Sea was determined. The mean prevalence values of NNV were higher than those found in wild fish populations in previous studies elsewhere in the Atlanto-Mediterranean region, where a atic Sea  and 19.6atic Sea . Howeveratic Sea . In the  in 2005 , and sin in 2005 . As for al niche ,34. Our al niche . Moleculal niche ,37. Afteal niche . Accordichanisms . Possibl species  and the  species ,40 have  species . However species . In thisat 20 °C . There iThe phylogenetic analysis of NNV across the four wild fish species from the Mediterranean Levantine Basin showed that all belonged to the RGNNV genotype. These results are in agreement with previous studies that reported the presence of RGNNV genotype only and revealed no reassortant genotypes in wild species ,42,43,44Lymphocystis Disease Virus (LCDV), which belongs to the family Iridoviridae. Iridoviruses possess a 20-sided icosahedron with a DNA core, and the particle is relatively large, measuring from 120–300 nm in diameter. The Lymphocystis virus causes a self-limiting disease in young fish [Lymphocystis disease was described in 2008, affecting cultured gilthead seabream (Sparus aurata) in Eilat land-based nursery tanks (Red Sea) [Lymphocystis disease was reported in reared gilthead seabream (Sparus aurata) along the Tunisian coast [Another viral disease that was detected in both the Mediterranean and Red Seas is ung fish ,46. An oRed Sea) . Later, N. randalli, and suggests that viral resistance should be considered as an additional mechanism that may favor the establishment of invasive species in their new environment. While the factors that influence effective establishment are still poorly understood [This study reported the high prevalence of NNV in the Lessepsian species derstood ,16, it iderstood ,17. Can


Objective: Epidemiologic studies investigating the association between dietary carbohydrates as well as glycemic index and glycemic load  and bladder cancer risk have yielded inconsistent results. The aim of the present meta-analysis is to summarize the evidence on this association.Materials and Methods: A comprehensive literature search of articles published by December 2019 was performed in PubMed, Scopus, and Web of Science databases. A random-effects model was used to calculate the pooled odds ratios (ORs) and their corresponding 95% confidence intervals (CIs).Results: Twelve observational studies were included in the final analysis. There was no evidence of an association between consumption of carbohydrates and bladder cancer risk . No statistically significant association between glycemic load and bladder cancer was likewise found . However, there was a significant positive association between glycemic index and bladder cancer risk . In the dose–response analysis, the pooled OR (95% CI) per 10 units of glycemic index per day was 1.02 .Conclusion: In this meta-analysis, glycemic index showed a positive linear association with bladder cancer risk. Bladder cancer is a common human malignancy, with an estimated 429,800 new cancer cases and 165,100 deaths in 2012 worldwide . EmerginCarbohydrates are one of the main dietary components affecting an individual's insulin secretion and blood glycemic response . GlycemiA comprehensive literature search of articles published by December 2019 was performed in PubMed, Scopus, and Web of Science databases with the following search algorithm: (diet or dietary or nutrition or nutrient or food or glycemic index or glycemic load or glycemic index or glycemic load or carbohydrate or carbohydrates) and  and  and (cohort or case–cohort or case–control or case control). We also examined the reference lists from retrieved articles and other relevant reviews to identify any additional studies. This study was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement .The inclusion criteria were as follows: (1) investigation of the association between dietary carbohydrates, glycemic index, or glycemic load and bladder cancer risk, (2) use of the following study design: case–control, nested case–control, cohort, and case–cohort studies as well as follow-up studies of randomized clinical trials, and (3) reporting of fully adjusted risk estimates, including relative risk (RR), hazard ratio (HR), or odds ratio (OR), and the corresponding 95% confidence intervals (CIs) for the association between bladder cancer and dietary carbohydrate, glycemic index, or glycemic load. If multiple publications used the same study population, the latest publication providing the largest sample size was included. The Participants, Intervention Comparators, Outcomes, Study Design criteria are presented in The following data were extracted by two independent investigators (HZ and XX): first author's surname, publication year, country where the study was performed, specific characteristics of the study population, study design, age of participants, study size, number of cases, duration of follow-up for prospective studies, dietary assessment method, exposure , quantity of intake, risk estimates, 95% CIs from the most fully adjusted models, and matched or adjusted variables .http://www.ohri.ca/programs/clinical_epidemiology/oxford.asp). Any disagreements were resolved by discussing with a third reviewer. NOS is an eight-item instrument that allows for the assessment of the study population and selection, study comparability, follow-up, and the outcome of interest. The range of possible scores is 0 to 9. Scores <7 and ≥7 were assigned for low- and high-quality studies, respectively.The quality of each study was assessed by the same two investigators (HZ and XX) using the Newcastle–Ottawa Scale (NOS)  and the risk of bladder cancer were examined by using a random-effects model . The posThe linear dose–response trends were computed from the natural logarithm of the ORs and 95% CIs across the categories of exposure (at least three categories) using the method proposed by Greenland and Longnecker . In addiQ and I2 statistics, with the significant level set at P ≤ 0.1 .Heterogeneity between studies was explored using  P ≤ 0.1 . Publica P ≤ 0.1  and Egge P ≤ 0.1 . A two-tThe process of literature search and selection is presented in I2 = 0.0% and Pheterogeneity = 0.618). In the subgroup analyses, neither geographic region nor study design modified the association between carbohydrate intake and bladder cancer risk . There was no evidence of differences by geographic area and study design . No difference was observed when data were stratified by geographic area and study design (Four studies –14 were y design . Leave-oy design .Pnonlinearity = 0.389). The curve showed a significant increase in bladder cancer risk with increasing units of glycemic index. The pooled OR (95% CI) per 10 units of glycemic index per day was 1.02  (We then further explored if there was a dose–response relation between dietary glycemic index and bladder cancer risk. There was no evidence of a non-linear association between glycemic index and bladder cancer risk (01–1.04) .In this meta-analysis of observational studies, we found that a higher glycemic index was significantly related to a greater risk of bladder cancer. No association was observed between risk of bladder cancer and carbohydrate intake or glycemic load. To the best of our knowledge, this is the first meta-analysis to evaluate the association between dietary carbohydrates, glycemic index, or glycemic load and bladder cancer risk.Recently, the relationship between carbohydrates, glycemic index, or glycemic load and the risk of human cancers has been widely studied. Schlesinger et al.  and MullOur study indicated that there was a positive linear association between glycemic index and bladder cancer risk. Glycemic index is an indicator of the quality of carbohydrate foods. Foods with high glycemic index are rapidly digested and lead to a rapid peak in blood glucose and the secretion of insulin . HyperinThis meta-analysis has several strengths. First, both high- and low-level analyses and dose–response analysis were performed for the association between glycemic index and bladder cancer risk, with consistent results. Second, there was no evidence of obvious heterogeneity across studies. Third, we performed a throughout literature search, and no publication bias was detected. Fourth, leave-one-out sensitivity analyses showed no marked difference in results, indicating that the pooled risk estimates were robust and not driven by any single study.As with any study, this meta-analysis also has some limitations. First, individuals with a diet of high carbohydrate intake, glycemic index, or glycemic load may also have other unhealthy behavioral and dietary habits and metabolic-related illness, such as lack of physical activity, excess intake of total energy, diabetes, and obesity, which may confound the true association. Second, measurement error of diet may exist. Food frequency questionnaires are not originally designed to assess glycemic index and glycemic load. Most dietary questionnaires have calculated the glycemic index or glycemic load values according to a limited number of food items, which may affect the accuracy of exposure assessment. Finally, the inclusion of case–control studies may lead to recall and selection bias.In conclusion, in this meta-analysis, glycemic index showed a positive linear association with bladder cancer risk. Carbohydrate intake and glycemic load were not associated with the risk of bladder cancer. Furthermore, large prospective studies that account for other relevant factors, such as body weight, diabetes, physical activity, and intake of total energy, are needed.The datasets generated for this study are available on request to the corresponding author.XX and BL contributed to the conception or design of the work. HZ, QM, and HS contributed to the acquisition, analysis, or interpretation of data for the work. XX and SW drafted the manuscript. HZ and BL critically revised the manuscript. All authors gave final approval and agree to be accountable for all aspects of work ensuring integrity and accuracy.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. The handling editor declared a shared affiliation with the authors at time of review.