Nut types tested included almonds, peanuts, pine nuts, pistachios and walnuts, with dose and length of supplementation varying greatly across studies. Most studies (n = 13) included individuals at risk for CVD, according to various criteria. Findings were inconsistent, with ten studies reporting no significant changes in vascular function and six studies (one acute and five chronic studies) reporting improvements in at least one measure of vascular function. In summary, nuts have the potential to improve vascular function and future studies should consider the population, dose and length of nut supplementation as well as suitability of the different vascular function techniques.Vascular stiffness can be measured using numerous techniques including assessments of central haemodynamics, aortic arterial stiffness, and indices of aortic wave reflection and endothelial dilatation. Impaired vascular function is associated with increased risk of cardiovascular disease (CVD). Epidemiological studies indicate that regular nut consumption reduces CVD risk, with one of the proposed mechanisms being via improvements in vascular function. This narrative review summarizes the evidence from a systematic search of the literature of the effects of tree nut and peanut consumption on measures of vascular function excluding flow mediated dilatation. A total of 16 studies were identified, with a mix of acute controlled studies ( The nutrient profile of both tree nuts and peanuts has been proposed to be beneficial for cardiovascular health . Nuts arNuts have positive effects on vascular endothelial function, with regular nut consumption associated with improved vasodilation (or reactivity) measured by flow mediated dilation (FMD) without affecting endothelium-independent vasodilatation ,6. WhileVascular reactivity is also measured using the device EndoPAT, which detects plethysmographic pressure changes in the finger tips caused by the arterial pulse, which are translated to a peripheral arterial tone (PAT) . EndoPATVascular stiffness pathology is complex and the techniques used to assess this vary in their accuracy, precision, sensitivity and specificity. Along the arterial tree there are differences in muscularity and elasticity, hence local vascular measures are considered to be more quantitative and sensitive than systemic indices . Whilst Currently there has been no systematic evaluation of the literature related to the effects of nuts on markers of vascular function other than FMD ,21. TherOnly peer-reviewed intervention studies published in English evaluating at least one type of nut (tree nuts and/or peanuts) and their effects on vascular function measured as either vascular stiffness or vascular reactivity (other than FMD) were included.All acute  and chronic  intervention studies were considered.Adult participants aged ≥18 years were considered.All types of tree nuts and peanuts were considered. Soy nuts were not included.Both controlled and uncontrolled studies were considered.Only noninvasive techniques used to assess vascular function were included, such as pulse wave velocity (PWV), pulse wave analysis (PWA), digital volume pulse (DVP), impedance cardiography and peripheral arterial tone via plethysmography . The outcome measures derived from these techniques are listed in In August 2017, the following databases were systematically searched for relevant articles: Medline, CINAHL, Scopus, Cochrane and Embase. The reference lists of relevant review articles found in the search were also manually checked for studies missed by the database searches. This search was updated in July 2018 with no additional relevant papers identified.Search terms were collated into two key groups: nuts and vascular function. The following terms for nuts were searched in all databases: ‘nut or nuts or almond* or pistachio* or hazelnut* or walnut* or cashew* or macadamia* or pecan* or peanut* or “Corylus avellana” or “Prunus dulcis” or “Prunus amygdalus” or “Pistacia vera” or pistacia or juglans) or “Anacardium occidentale” or “Carya illinoinensis” or “Arachis hypogaea” or groundnut*’. The second group of terms were based on vascular function: “Arter* stiff*” or “Arter* complian*” or “arter* elasticit*” or “aortic stiffness*” or “vascular* stiff*” or “aortic complian*” or “aortic elasticit*” or “vascular* complian*” or “vascular* elasticit*” or “Arter* pressur*” or “aortic pressur*” or “vascular* pressur*” or laei OR saei or “pulse wave*” or “reflective index” or “Beta stiffness index” or “Augmentation Index” or “elasticity index” or “endopat” or “ankle brachial index” or “vascular resistance” or “systemic vascular resistance” or “peripheral resistance” or “arterial pressure wave” or “endothelial function”. The two groups were combined to search for relevant studies.After removal of duplicates, studies were screened by title and abstract. The full text of all articles was then reviewed before final inclusion in the review.Extracted data from all studies included study design, sample size, participant characteristics , sex and health status), the dietary intervention including the amount and type of nut, duration, placebo or control conditions, fed or fasted state for assessments and vascular function outcome.n = 13) [n = 3) [n = 4) [n = 3) [n = 1) [The studies included in the review comprise three acute controlled studies, one uncontrolled pre/post chronic study, seven controlled chronic crossover studies and five controlled parallel studies. The majority of studies were conducted in middle-aged participants who were overweight or obese. Most studies included both male and female participants; Kasliwal et al. 2015 had a higher proportion of males compared to females. Three studies used only males ,23,24 ann = 13) ,35,36,37 [n = 3) ,30,37, e [n = 4) ,29, type [n = 3) ,33,36 or [n = 1) .n = 6), PWV (n = 3), small and large arterial elasticity (n = 1) and total peripheral resistance (TPR) (n = 2). Reactive Hyperemia Index (RHI), a measure of vascular reactivity, was reported in nine studies.A range of outcome measures were reported in the studies . The outn = 4) [n = 3) [The most frequently evaluated nut type was walnuts, used in six out of the 16 studies ,26,27,31n = 4) ,29,32,33 [n = 3) ,34,36, w [n = 3) , and two [n = 3) ,37.p = 0.01) for RHI (vascular reactivity) but not for AI (vascular stiffness). They also reported that walnut oil favourably improved vascular reactivity compared to walnut skins.Of the 16 studies, three evaluated the post-prandial effects of a single meal containing nuts on vascular function. The post-prandial duration ranged from 1–8 h. Two studies compared meals containing nuts against control meals; one study compared muffins containing nuts against muffins with sunflower oil . The othp = 0.06) from 20% of total energy intake from almonds for 24 weeks.Gulati et al.  used a tThe majority of chronic studies included in this review used a cross-over design (8 out of 13). Three studies assessed vascular reactivity using EndoPAT (RHI) but no significant effects of nuts were reported ,33,34. Tp = 0.008, effect size = 0.04). There was no improvement in AI after 15 g/day of walnuts [−5, p < 0.0001) from 63–126 g/day of pistachios for 4 weeks. Sauder et al. [p = 0.004). West et al. 2010 [p ≤ 0.05) from 37 g/day of walnuts for 6 weeks.Vascular stiffness was assessed in seven studies. Barbour et al.  reported walnuts  or 59–12 walnuts  for 4 we walnuts  reportedr et al.  also repal. 2010  reportedA range of control diets were administered, including habitual diets ,35, modep = 0.025) with 40 g/day of walnuts compared to a control diet of 5 g/day of walnuts for 4 weeks and also a better post-prandial response in the 40 g/day group. However, Lopez-Uriarte et al. [p = 0.01 and 0.037, respectively) from 40 g/day of pistachios for 4 weeks.Five chronic studies used a parallel study design. These studies ranged in duration from 4 to 12 weeks and included nut doses of 28 g to 40 g per day. The majority of studies (four out of five) assessed changes in vascular function using Endo-PAT with mixed results. Holt et al.  reportede et al.  and Djoue et al.  reportede et al.  reportede et al.  had a sme et al.  and Lee e et al.  used a le et al.  reportedWhile the control diets differed among studies, the majority included non-nut groups. Djousse et al.  comparedThis review found that studies evaluating the effect of tree nut and peanut consumption on vascular function have shown mixed effects, with six out of 16 studies reporting a benefit associated with nut consumption and ten showing no effect. Acute crossover studies failed to demonstrate that nut consumption can affect vascular reactivity or vascular stiffness post-prandially. Although acute vascular reactivity was improved with consumption of walnut oil, this was not compared to a control group. Evidence from chronic studies suggest that regular consumption of nuts can improve TPR, with all three studies that assessed this outcome reporting a positive effect, with two of these studies assessing the effects of pistachios and one the effects of walnuts. Improved arterial stiffness was also observed following peanut and pistachio consumption compared to a nut-free diet and lifestyle modification, respectively. Only one study in walnuts observed improvements in vascular reactivity. Therefore, while there is evidence from some studies that nuts can improve arterial function, the findings are inconsistent.When considering the effects of nuts on vascular function the population type is important. The studies in healthy populations did not find significant improvements, whereas six (out of the 12) studies in people with CVD risk factors reported significant changes in at least one measure of vascular function ,32,33,35A similarity between this review and that performed by both Xiao et al.  and NealIn comparing the findings in this review and with previous reviews focused on FMD outcomes, both the doses of nuts provided and length of supplementation should be considered. In the current review, the range of nut doses (15–128 g/day) is lower compared to the doses used in the studies analysed by Xiao et al.  (52–128 Furthermore, inconsistency in results may be due to differences in study durations. The duration of the studies included by Xiao et al.  ranged fThis purpose of this review was to summarise the evidence for improved vascular stiffness or reactivity following consumption of tree nuts and peanuts. Based on the studies included in this review, there is some evidence for an improvement in vascular stiffness, with 5 of 13 chronic studies reporting significant reductions following nut consumption. Unfortunately, the small number of studies, along with the diverse range of outcome measures, nut types evaluated and duration of interventions limits our ability to draw a definitive conclusion. Similar to recommendations from other recent reviews in FMD, we emphasise the need for more nut intervention studies assessing functional measures of vascular health.


To date, type 2 diabetes is considered to be a “bi-hormonal disorder” rather than an “insulin-centric disorder,” suggesting that glucagon is as important as insulin. Although glucagon increases hepatic glucose production and blood glucose levels, paradoxical glucagon hypersecretion is observed in diabetes. Recently, insulin resistance in pancreatic α cells has been proposed to be associated with glucagon dysregulation. Moreover, cell autonomous dysfunction of α cells is involved in the etiology of diabetes. In this review, we summarize the current knowledge about the physiological and pathological roles of glucagon. Neuronal and hormonal factors in the body strictly control blood glucose levels. The development of diabetes is attributed to the breakdown of the blood glucose control mechanism . In 1921In 1923, Kimball and Murlin discovered glucagon from pancreas . GlucagoType 2 diabetes is caused by relative insulin hyposecretion and insulin resistance. In particular, decreased insulin secretion due to reduced pancreatic β cell mass is considered the leading cause of the onset and exacerbation of diabetes. However, in recent years, it has been surprisingly reported that the administration of streptozotocin (STZ), which destroys β cells and completely inhibits insulin secretion, does not increase blood glucose levels in glucagon deficient mice . When STIn vivo, glucagon secretion from the pancreatic α cells is strictly controlled by insulin ,15. FurtATP channels and reduces glucagon secretion [ATP channels resulting in the influx of Na+ from Na+ channels, followed by Ca2+ influx via P/Q-type Ca2+ channels [2+ is the main trigger of glucagon secretion, under conditions where glucose strongly suppresses glucagon secretion in isolated islets, the Ca2+ signal is only moderately or temporarily decreased [2+ signaling [2+ and cAMP are only sufficient regulators for glucagon secretion. Under high glucose conditions, an SGLT-dependent increase in the intracellular Na+ concentration and a decrease in the mitochondrial pH increase glucagon secretion via the KATP channels [ATP channels, such as insulin secretion and the cAMP-PKA signal. However, such a result must be validated in future studies.When the blood glucose level deviates from the normal level, either hypoglycemia or hyperglycemia occurs. When healthy individuals are subjected to fasting, insulin secretion from β cells is suppressed, whereas glucagon secretion from α cells is enhanced. By contrast, opposite changes are expected to occur when healthy individuals are fed. However, in patients with diabetes, a paradoxical glucagon hypersecretion occurs despite the presence of hyperglycemia. The most important factor affecting glucagon secretion in healthy individuals is glucose per se. Several glucose transporters, including GLUT1, sodium glucose co-transporter (SGLT1), and SGLT2, are expressed in human pancreatic α cells ,17,18,19ecretion . ATP depchannels . Althougecreased . In addiignaling . That ischannels . These pIn diabetes, paradoxical glucagon hypersecretion occurs despite the presence of hyperglycemia, and glucagon hyposecretion occurs despite the presence of hypoglycemia ,27,28. T2+ channel [ATP channels is reduced via PI3K, which is a signal molecule downstream of IR, to suppress glucagon secretion [In general, insulin resistance is a condition in which classical insulin target organs have decreased insulin sensitivity resulting in decrease of glucose uptake . Increas channel . This me channel . Insulin channel . In addiecretion ,35. In aecretion . Therefoproglucagon gene, is enhanced via mitogen-activated protein kinase (P-38) and extracellular signal-regulated kinase (ERK) 44/42 pathways [In diabetic state, insulin resistance might exist in pancreatic α cells. If insulin resistance is present in pancreatic α cells, these insulin signals are attenuated and affect glucagon secretion. In fact, studies of pancreatic α-cell-specific IR knockout mice have shown increased glucose and plasma glucagon levels ad libitum fed . Furtherpathways . These rpathways , which ipathways . In addiParacrine signaling indicates that secretion from cells does not act on distant cells through the general circulation but acts on adjacent cells via direct diffusion and other similar mechanisms. In rodent islets, β cells are found in the center of the islet and α cells around the islets. Anatomically, paracrine factors from β cells may affect α cells, considering that blood flows from the central to the peripheral part in the islet . By cont2+, and insulin secreted by pancreatic β cells suppress glucagon secretion from α cells. GABA is known as a major inhibitory transmitter in the central nervous system, but has also been shown to be present with high concentrations in the pancreas [2+ concentration is increased. In mouse islets and α cell lines (α-TC1-9), GABA released from β cells binds to the GABA-A receptors of α cells and suppresses glucagon secretion [2+ is contained in the insulin granules of pancreatic β cells. In the perfused pancreas of rats, glucose-induced Zn2+ secretion from β cells suppressed glucagon secretion [2+ did not suppress glucagon secretion [2+ granule transporter knockout mice [2+ in regulating glucagon secretion remains controversial. In addition, glucagon exocytosis in α cells is inhibited by juxtacrine via the Ephin subtype A (EphA) of β cells and EphA 4/7 receptor of α cells [Just as insulin from β cells suppresses glucagon secretion from α cells, glucagon secretion from pancreatic α cells is autocrinally and paracrinally regulated by various factors secreted by the pancreatic islet α, β, and δ cells. GABA, Znpancreas ,45,46. Iecretion ,48,49. Aecretion . Howeverecretion . Furtherout mice . Therefo α cells . That is α cells ,55. Soma α cells ,56,57. M α cells . Somatos α cells . Moreove α cells . In addi α cells . That is α cells ,63,64. W α cells . Glucago α cells . TherefoCurrently, there are several different diabetes drugs available worldwide, some of which affect glucagon in various ways and exhibit hypoglycemic action. Glucagon-like peptide-1 (GLP-1) agonist and dipeptidyl peptidase-4 (DPP4) inhibitor, which are incretin-related drugs, suppress glucagon secretion. DPP4 is an enzyme that inhibits GLP-1. When either of these drugs is administered, the concentration of GLP-1 in the blood increases. Administration of GLP-1 suppresses glucagon secretion in patients with type 2 diabetes, as measured using a glucose clamp test . The mecIn addition, glucagon receptor antagonists that suppress gluconeogenesis in the liver and lower blood glucose levels have recently been developed. LY2409021 is a low molecular weight compound that is an antagonist to the glucagon receptor. A marked improvement in blood glucose was observed after 12 weeks of administration of this compound to patients with type 2 diabetes . HoweverAmong various antidiabetic drugs currently available, incretin-related drugs and biguanide are considered to exert a glucagon inhibitory effect, and their mechanism of action has been elucidated. Furthermore, new drugs including glucagon receptor antagonists and neutralizing antibodies are currently under development, and the importance of glucagon in diabetes is increasingly recognized.Studies regarding to the glucagon are few compared to those of insulin, and there are two major reasons. First, there is a problem with the glucagon assay system. Conventionally, glucagon is measured by using radioimmunoassay methods with antibodies that recognize the C-terminal structure of glucagon. However, these methods cannot accurately measure glucagon because of the existence of glucagon-like peptides, whose amino acids sequence overlap with glucagon. In particular, glicentin, the C-terminal sequence of which is identical to that of glucagon, causes cross-reaction. However, in recent years, glucagon can be more accurately measured by newly developed methods, i.e., sandwich enzyme-linked immunosorbent assay using antibodies that recognize the C-terminus and N-terminus of glucagon, and liquid chromatography/mass spectrometry (LC/MS/MS). Therefore, the problem seems to be solved (but this is not the case). Second, there is a problem with the material used. In the case of mice and rats used in the experiment, islets cells are mainly composed of β cells and α cells by around 80% and 10%, respectively. Therefore, α cell research is challenging because the number of α cells is much fewer than β cells. In addition, although insulin is a relatively stable hormone, glucagon is unstable and rapidly broken down at room temperature. Moreover, most studies analyzing glucagon secretion have used transformed α cell lines, and only a few studies have utilized native α cells, which is attributed to the absence of established methods that isolate pancreatic α cells owing to their unstable viability. Currently, the commonly used α cell lines are InR1G9 , HIT T15In type 2 diabetes, glucagon dysregulation is as important as insulin dysregulation. In fact, paradoxical glucagon hypersecretion under hyperglycemic conditions and impaired glucagon secretion under hypoglycemic conditions are commonly observed in diabetes. Therefore, the relationship between insulin and glucagon—the major factors that determine blood glucose level—is important in diabetes research. In recent years, insulin resistance in pancreatic α cells has been proposed; moreover, the relationship between insulin and glucagon has been gaining considerable attention. In insulin-resistant α-cell lines, the regulation of glucagon secretion and expression is disturbed. In pancreatic α cell-specific IR knockout mice, increased postprandial plasma glucagon levels and hyperglycemia have been observed. Therefore, insulin resistance in α cells is associated with glucagon dysregulation in diabetes. Although the mechanism underlying cell autonomous dysfunction of α cells has not yet been elucidated, it is believed to be correlated with the pathophysiology of diabetes. Therefore, α cell research using native α cells or α cell lines with similar properties to native α cells should be performed for better understanding of the pathophysiology of diabetes.Type 2 diabetes is considered to be a “bi-hormonal disorder” rather than an “insulin-centric disorder,” suggesting that glucagon is as important as insulin. Recently, “glucagonocentric hypothesis”, in which glucagon contributes more to increase in blood sugar level than insulin, has attracted attention.Insulin resistance in pancreatic α cells means a state in which the insulin signal of α cells is attenuated. In normal conditions, insulin suppresses the secretion of glucagon from pancreatic α cells. However, when insulin resistance exists in diabetic pancreatic α cells, insulin can no longer suppress glucagon secretion from α cells, which results in hypersecretion of glucagon. Hypothetical paradoxical glucagon hypersecretion observed in diabetes.ATP channel has been reported.The molecular mechanism of glucose-regulated glucagon secretion in pancreatic α-cells has not yet been elucidated, but the involvement of cAMP, ATP/ADP ratio, and KThe new drugs such as glucagon receptor antagonist and neutralizing antibody are currently developing.Glucagon studies have problems with measurement systems and materials. The problem of the measurement system is being solved, but the problem of the material has not been solved yet. Therefore, the establishment of new methods for isolating native α cells or construction of a new α cell line having similar properties with native α cells is required for future α cell research.Here are the final summaries;


Escherichia coli cell lineages exhibit comparable patterns of mortality and immortality. Through single-cell microscopy and microfluidic techniques, we find that these patterns are explained by the dynamics of damage accumulation and asymmetric partitioning between daughter cells. At low damage accumulation rates, both aging and rejuvenating lineages retain immortality by reaching their respective states of physiological equilibrium. We show that both asymmetry and equilibrium are present in repair mutants lacking certain repair chaperones, suggesting that intact repair capacity is not essential for immortal proliferation. We show that this growth equilibrium, however, is displaced by extrinsic damage in a dosage-dependent response. Moreover, we demonstrate that aging lineages become mortal when damage accumulation rates surpass a threshold, whereas rejuvenating lineages within the same population remain immortal. Thus, the processes of damage accumulation and partitioning through asymmetric cell division are essential in the determination of proliferative mortality and immortality in bacterial populations. This study provides further evidence for the characterization of cellular aging as a general process, affecting prokaryotes and eukaryotes alike and according to similar evolutionary constraints.Cellular aging, a progressive functional decline driven by damage accumulation, often culminates in the mortality of a cell lineage. Certain lineages, however, are able to sustain long-lasting immortality, as prominently exemplified by stem cells. Here, we show that  A study of Escherichia coli shows that bacterial lineages maintain replicative immortality by reaching an equilibrium between aging and rejuvenation; when this equilibrium is disrupted, aging lineages cross their immortality threshold, becoming mortal, while rejuvenating lineages are favored by asymmetry and retain immortality within the same population. Aging, or the progressive loss of function at the macromolecule, tissue, organ, or individual level, is largely driven by the deterioration of intracellular processes. Accordingly, the hallmarks of the aging phenotype—such as telomeric attrition, mitochondrial dysfunction, loss of proteostasis, and genomic instability—which have been well characterized by previous studies , reveal The cellular aging process encompasses both multi- and unicellular organisms, such as yeast, diatoms, and even bacteria –8. Due tBesides aggregating and repairing damaged components, bacterial populations have developed another remarkable strategy to handle nongenetic damage. Experimental data from long-term microscopy of bacterial lineages revealed that, in the presence of intracellular damage, each cellular division produces 2 physiologically asymmetric daughters ,11,15–17E. coli lineages depends on the physiological equilibrium produced by contrasting damage accumulation and asymmetric partitioning. We demonstrate that unstressed lineages accumulate damage produced by standard respiration levels, subsequently partitioning this load with a level of asymmetry that allows for the dilution of damage within both new and old daughters. We show that E. coli mutants with decreased repair capacity also exhibit asymmetric new and old daughters, reaching distinct states of growth equilibrium. Furthermore, bacterial aging responds with a positive dosage relationship to an external damaging agent, which progressively disrupts proteostasis by increasing damage accumulation rates and disrupting asymmetry. With a sufficiently elevated stress level, the damage accumulation within old lineages surpasses their immortality threshold, leading these lineages to arrest division and become mortal. However, due to asymmetric partitioning, new lineages within the same population retain proliferative immortality. Our results show that the appropriate model and system can contribute to identifying the dynamics of mortality and immortality in the context of cellular aging.A better understanding of how the key features of aging are interconnected requires the eventual development of conceptual and mathematical models that can integrate with experimental studies the growth and aging of individual organisms or cells. Unicellular systems, such as bacteria, satisfy all these requirements. Here, we show that the maintenance of proliferative immortality in E. coli cells using microfluidic devices. We employed the “mother machine” design ) daughter subpopulations emerged  was significantly larger than zero . These results suggest that old daughters in our populations are inheriting a larger damage load upon division, despite the absence of extrinsic damage in our growth conditions.Under such conditions, our previous studies have shown that new and old daughters display physiological asymmetry and long-term growth stability . We conf emerged . In thesT1 and T2, reconstructing the progression of aging within a lineage. For this goal, we described cell lineage dynamics though 3 key parameters: Π, the doubling time of a damage-free cell; λ, the rate of damage accumulation within a single cell (0 to approximately 0.01 min−1); and a, the partitioning asymmetry, ranging from 0 (complete asymmetry) to 0.5 (symmetric division).To quantify the possible damage accumulation and partitioning in these populations, we applied these results to a population genetics model on unicellular aging . BecauseE. coli . These longer doubling times were driven by damage accumulation—which occurred at an average rate λ = 0.0028 min−1—thus suggesting that metabolic processes in healthy cells may induce the retention of intrinsic damage. Finally, as suggested by the separation between T1 and T2 subpopulations, we verified that these damage loads were partitioned asymmetrically at division, with old daughters inheriting 63% of the maternal damage (a = 0.37).Our growth parameters revealed the presence of intrinsic damage and asymmetry in physiologically stable E. coli . Despitea can be used to predict doubling times T1 and T2. In T0 = T1 or T0 = T2  and T2 . In the stable environment of microfluidic devices, this equilibrium can still be disrupted by the stochasticity present in doubling times. This stochasticity can be described as random variables ξ1 acting on the slopes in Ti = T0 × (a + ξ1) + b, obtained in each generation from a Gaussian distribution with SD of σ1. Loss of equilibrium occurs when a2 + σ12 ≥ 1 (see 1 for T1 and T2 lines by obtaining the deviations from slopes in (Ti − b) ÷ T0 = a + σ1. Both new (σ1 = 0.0657) and old (σ1 = 0.0876) lineages satisfied the stability requirement a2 + σ12 < 1, with a2 + σ12 = 0.0649 for T1 and a2 + σ12 = 0.1032 for T2.To confirm the long-term stability of new and old lineages in our experiments, we analyzed linear regressions between escribed  S1 Fig)T0 and T1−1, would progressively drive the equilibrium points toward longer doubling times. Because asymmetric partitioning produces higher doubling times in old daughters, a sufficiently intense λ would act as a differential mortality threshold, leading to division arrest—i.e., a state of mortality—in the old lineage, whereas new daughters remain immortal  and division . Furthermore, we compared each mother to its old daughter and verified that k2 = k0 . This indicates that old lineages in a state of equilibrium, as observed in the mother machine, are born with a constant level of intrinsic damage. Consequently, the damage accumulated by a mother over its lifetime is equivalent to the load inherited by new daughters upon division, or λ × T0 = k1 .To connect our observation of immortality and growth equilibrium to the internal dynamics of damage accumulation and partitioning, we estimated damage loads using our growth parameters. From experimental doubling times , we calck1 = D0 × a = λ × T0), whereas old daughters inherit the same amount the mother had at birth (k2 = D0× (1 − a) = k0). These dynamics of damage accumulation and partitioning allow for a state of physiological equilibrium, in which old lineages display stable growth over time and retain proliferative immortality.Taken together, our results suggest that unstressed bacterial populations accumulate intrinsic nongenetic damage. Every generation, new daughters inherit the damage a mother accumulated over its lifetime  fusioned to yellow fluorescent protein (YFP). IbpA-YFP was shown to colocalize with protein aggregates in bacterial cells ) siblings . A distinct pattern emerged from the analysis of ΔdnaK mutants, with several mortality events occurring over time .Given the asymmetric damage partitioning in equilibrium lineages, we investigated the relevance of the protein repair machinery for the maintenance of proliferative immortality. For this, we employed gregates ,12,27. Wperiment . We obsever time . NonetheΔclpB and ΔdnaK populations, we analyzed the linear models presented in 1 in (Ti − b) ÷ T0 = a + σ1. To generate 95% confidence intervals (CIs), we performed a 10,000-fold bootstrap on T0, T1, and T2 trios. We verified that ΔclpB mutants satisfied the stability requirement a2 + σ12 < 1  and old lineages (a2 + σ12 = 0.107 [0.051–0.206]). For ΔdnaK, although new (a2 + σ12 = 0.241 [0.020–0.624]) and old lineages (a2 + σ12 = 0.473 [0.113–0.956]) satisfied the stability requirement, several mortality events were observed, with our bootstrap analysis suggesting a 1.45% probability of equilibrium loss  in ΔdnaK old lineages.To verify the stability of growth equilibrium in asymmetric  σ12 < 1 , thus reΔclpB mutants still allowed for the stable proliferation of new and old daughters, old lineages in ΔdnaK mutants begin to show signs of stability loss. We therefore hypothesize that dynamics of damage accumulation may greatly impact proliferative immortality, with asymmetry determining a differential fate for new and old lineages.Taken together, these results suggest that repair chaperones ClpB and DnaK might have distinct roles in the maintenance of equilibrium stability. Although the decreased protein repair capacity in To investigate whether aging cell lineages would retain physiological equilibrium—therefore proliferative immortality—under increasing levels of damage accumulation, we cultured bacteria in the presence of extrinsic damage. We employed light excitation (490 nm wavelength), commonly used for green-fluorescent protein imaging, as a damaging agent known for inducing the production of reactive oxygen species and mitochondrial damage ,28–30. Bt tests; p < 0.001). The treatments revealed a significant effect of both age and light exposure on individual elongation rates, with new daughters maintaining significantly faster growth than their old siblings in all cases  was significantly reduced at 3 s of exposure, when compared to control conditions . This outcome was surprising, because one of the advantages of asymmetric damage partitioning in bacterial populations is the ability to endure higher levels of damage  ÷ 2) and deterministic (D2 ÷ 4) components. Values presented as mean and 95% CIs. CI, confidence interval.Partitioning of doubling time variances into stochastic ([V(XLSX)Click here for additional data file.S1 DataDoubling times of unstressed wild-type populations as trios composed of a mother bacterium and its 2 daughters.(XLSX)Click here for additional data file.S2 DataΔclpB trios comprising a mother and its 2 daughters. ClpB, chaperone protein ClpB.Data for (XLSX)Click here for additional data file.S3 DataΔdnaK trios comprising a mother and its 2 daughters. DnaK, chaperone protein DnaK.Data for (CSV)Click here for additional data file.S4 DataData for wild-type trios comprising a mother and its 2 daughters, exposed to 70, 700, 1,000, 1,500, or 3,000 ms of light excitation.(XLSX)Click here for additional data file.S5 DataData for wild-type new and old daughters exposed to 38 °C, 40 °C, or 43 °C.(XLSX)Click here for additional data file.S6 DataData for wild-type new and old daughters exposed to 2, 4, or 5 μg/ml streptomycin.(XLSX)Click here for additional data file.S7 DataRaw data used for Figs (XLSX)Click here for additional data file.


Candidatus Liberibacter asiaticus.” Prior work using mapping-based approaches identified prophage-like sequences in some “Ca. Liberibacter asiaticus” genomes but not all. Here, we utilized a de novo approach that expands the number of prophage-like elements found in “Ca. Liberibacter asiaticus” from 16 to 33 and identified at least one prophage-like sequence in all “Ca. Liberibacter asiaticus” strains. Furthermore, we identified a prophage-like sequence type that is a remnant of an integrated prophage—expanding the number of prophage types in “Ca. Liberibacter asiaticus” from 3 to 4. Overall, the findings will help researchers investigate the role of prophage in the ecology, evolution, and pathogenicity of “Ca. Liberibacter asiaticus.”Huanglongbing (HLB) disease is threatening citrus production worldwide. The causative agent is “ Candidatus Liberibacter asiaticus” is an alphaproteobacterium of the Rhizobiaceae family that has been identified as the causative agent of HLB. The virulence of “Ca. Liberibacter asiaticus” has been attributed, in part, to prophage-carried genes. Prophage and prophage-like elements have been identified in 12 of the 15 available “Ca. Liberibacter asiaticus” genomes and are classified into three prophage types. Here, we reexamined all 15 “Ca. Liberibacter asiaticus” genomes using a de novo prediction approach and expanded the number of prophage-like elements from 16 to 33. Further, we found that all of the “Ca. Liberibacter asiaticus” genomes contained at least one prophage-like sequence. Comparative analysis revealed a prevalent, albeit previously unknown, prophage-like sequence type that is a remnant of an integrated prophage. Notably, this remnant prophage is found in the Ishi-1 “Ca. Liberibacter asiaticus” strain that had previously been reported as lacking prophages. Our findings provide both a resource for data and new insights into the evolutionary relationship between phage and “Ca. Liberibacter asiaticus” pathogenicity.Huanglongbing disease  is a severe worldwide infectious disease for citrus family plants. The pathogen “IMPORTANCE Huanglongbing (HLB) disease is threatening citrus production worldwide. The causative agent is “Candidatus Liberibacter asiaticus.” Prior work using mapping-based approaches identified prophage-like sequences in some “Ca. Liberibacter asiaticus” genomes but not all. Here, we utilized a de novo approach that expands the number of prophage-like elements found in “Ca. Liberibacter asiaticus” from 16 to 33 and identified at least one prophage-like sequence in all “Ca. Liberibacter asiaticus” strains. Furthermore, we identified a prophage-like sequence type that is a remnant of an integrated prophage—expanding the number of prophage types in “Ca. Liberibacter asiaticus” from 3 to 4. Overall, the findings will help researchers investigate the role of prophage in the ecology, evolution, and pathogenicity of “Ca. Liberibacter asiaticus.” Candidatus Liberibacter asiaticus” has been identified as one of the three “Ca. Liberibacter” species that cause Huanglongbing disease . HLB is a major threat to the worldwide citrus-growing industry  Genome database . They were discovered when the sequencing of a “Ca. Liberibacter asiaticus” strain showed circular contigs rich in phage open reading frames (ORFs) . We compared the predicted sequences to phage genomes available from NCBI  and coverage (C) values of two-way local alignments are presented on this table. Blue, mustard, and pink cells stand for high, intermediate, and low values, respectively . All sequences were classified as new prophage-like sequence type 4. Download Copyright © 2019 Dominguez-Mirazo et al.2019Dominguez-Mirazo et al.Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the YP_007011137.1) found only in prophages of type 2 and 3 (see Ca. Liberibacter asiaticus” strain AHCA1 harbors a type 3 prophage sequence. The truncation of the type 1 and type 3 prophage sequences and their closeness to the type 4 prophage-like sequence in strains FL17 and AHCA1 explain the incorrect concatenation of these sequences into a single prediction by Virsorter and PHASTER. The prophage fragment at the end of FL17-a (end of contig 1) joined to type 1 sequences FL17-b (contig 2) and FL17-c (contig 3) reconstructs the sequence of a type 1 prophage  Fig. 1,, but thend 3 see  suggest 10.1128/mSphere.00409-19.2TABLE S2Table S2, DOCX file, 0.2 MB.AHCA1-a gene function. ORFs predicted for AHCA1-a were annotated using BLASTp against the nonredundant database. Information includes gene start and end positions, UniRef ID, and predicted gene function. The corresponding locus_tag from the GenBank AHCA1 genome annotation is presented in the succeeding columns with the locus_tag ID, start and end positions in the bacterial genome, and annotated function. Note that some ORFs lack a corresponding locus_tag and that some locus_tags lack a corresponding ORF. Download Copyright © 2019 Dominguez-Mirazo et al.2019Dominguez-Mirazo et al.Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the In order to evaluate the robustness of the classification identified in prior sections, we analyzed the pan-genome content of prophages through a clustering approach . ClusterCa. Liberibacter asiaticus” strains, including those previously reported and the ones introduced in this study. There are 5 newly reported elements that belong to prophages of type 1, 2, or 3 (see “Prophage-like sequence identification” section below) and 12 type 4 prophage-like sequences. Our findings expand the number of known prophage elements in “Ca. Liberibacter asiaticus” from 16 to 33.Ca. Liberibacter asiaticus” prophage-like sequences, we functionally annotated the representative prophage-like sequence Ishi-1-a  resemble different parts of a portal protein found also in “Ca. Liberibacter solanacearum.” ORF18 and ORF19 are remnants of a DNA packaging protein similar to one present in “Candidatus Liberibacter africanus.” There are three ORFs involved in DNA replication: a gene that codes for a transcriptional regulator of the XRE family (ORF22) and two fragments (ORF23 and ORF24) of a helicase coding gene. Furthermore, no attachment sites were found in the bacterial genome near the type 4 sequences.In Ishi-1, the sequence of a head protein found in “The disruption of viral-particle-forming genes, along with the absence of att sites, the G+C content closely resembling that of the bacterial host, and the small sequence size, implies that type 4 sequences are remnants of a prophage integrated into the bacterial genome.Ca. Liberibacter asiaticus”-related species. The uncultivated “Candidatus Liberibacter africanus” and “Candidatus Liberibacter americanus” species are closely related to “Ca. Liberibacter asiaticus” and have been identified as less-prevalent causative agents of HLB (Ca. Liberibacter africanus” and “Ca. Liberibacter americanus” (Ca. Liberibacter asiaticus”/“Ca. Liberibacter africanus” and the “Ca. Liberibacter americanus” clades [Ca. Liberibacter africanus” and the Ishi-1 strain (81.60%) and in the ANI values determined for the corresponding predicted type 4 prophage-like sequences (81.69%). Similarly, the AAI values determined for “Ca. Liberibacter americanus” and “Ca. Liberibacter africanus,” for “Ca. Liberibacter americanus” and Ishi-1, and for “Ca. Liberibacter africanus” and Ishi-1  match the AAI values determined for the corresponding predicted type 4 prophage-like sequences . These findings are consistent with the integration of a type 4 prophage-like sequence prior to lineage divergence. The presence of a type 4 sequence in almost all evaluated genomes challenges the common view of “Ca. Liberibacter asiaticus” strains lacking prophage or prophage-like sequences.To assess the evolutionary origin of the type 4 prophage-like sequence and its time of insertion into the bacterial genome, we searched for type 4 sequences in “s of HLB . Accordiricanus” . The pre” clades ). To tesCa. Liberibacter asiaticus” genome has usually included analysis of prophage types  Nucleotide Database, under genome identifier (ID) 1750 (https://www.ncbi.nlm.nih.gov/genome/genomes/1750). All “Ca. Liberibacter asiaticus” prophages previously reported were also retrieved from NCBI  1750   was considered high with I values ≥ 95, intermediate with I values < 95 and ≥ 85, and low with I values < 85. Two sets of thresholds were used to evaluate the coverage. When the query sequence was shorter than the target, the coverage (C) was considered high with C values ≥ 90, intermediate with C values < 90 and ≥ 75, and low with C values < 75. When the query sequence was longer than the target, the coverage was considered high with C values ≥ 33, intermediate with C values < 33 and ≥ 25, and low with C values < 25. A sequence was classified as a particular phage type if none of its BLAST values were “low” and if it had the highest fraction of “high” scored values. If more than one phage type fulfilled the aforementioned conditions, the assigned phage type was associated with that with the highest identity value. The putative prophages that were not classified as type 1, 2, or 3 were aligned against each other as part of the process of identifying a potential (new) type 4 group.For sequence annotation, open reading frames (ORFs) were identified with GeneMarkS-2 . Data rehttps://github.com/WeitzGroup/CLas_prophages with zenodo link at https://doi.org/10.5281/zenodo.3405598.R (v3.3) and Bash were used to generate all figures. Code is available at


Background: Although the effects of mental fatigue on cognitive–motor function and psychological state in young adults are well-documented, its effects in the elderly are not completely understood. The aim of this study was to estimate the effect of prolonged cognitive load on the indicators of psychological, cognitive, and motor functions.Methods: Fifteen young and 15 elderly men were asked to perform a 2 h “Go/NoGo” task. Psychological state (mood and motivation), cognitive , and motor (motor cortex excitability and grip strength) functions were measured before and after the task. During the 2 h task, both groups had a significantly similar increase in the number of “Incorrect NoGo” errors. Only in young men reaction time (RT) of “Incorrect NoGo” and intraindividual variability of RT of “Incorrect NoGo” significantly increased during task. After the task, handgrip strength decreased for the young men, whereas latency of motor evoked potentials prolonged both groups. Nevertheless, both groups indicated that they felt fatigue after the 2 h task; we observed that mental demand increased, whereas intrinsic motivation and mood decreased only in young men. Prolonged task decreased the switching/rest ratio of oxygenated hemoglobin for the young and the elderly men; however, greater for elderly than young men. Interestingly, the more the prefrontal cortex was activated before the 2 h task during the switching task, the fewer of “Incorrect NoGo” errors made by the young men and the greater the number of errors made by the elderly men.Conclusion: Because of the greater mental load and (possibly) greater activation of prefrontal cortex during the 2 h “Go/NoGo” task, there was greater mental and neuromuscular performance fatigue in young men than in elderly men. Functional limitations, disabilities, mortality, and other adversative consequences in the elderly can be highly predicted by fatigue . Mental The self-control mechanism is highly dependent on the so-called executive function, which operates effectively if concentration on the required object and inhibition of unnecessary objects (temptations), working memory, and flexible switching of attention work well . The mai“Go/NoGo” and “Stroop” test exercises are widely used for inducing cognitive fatigue . These aThe speed of processing, working memory, inhibitory function, and long-term memory decline with age, similarly, the brain structure size and white matter integrity decrease . The ageThe main aim of our studies was to test the following hypothesis. As older adults more highly activate executive, cognitive, and association brain regions aiming at performing the tasks , (1) eld2] and 15 elderly  men. Young and elderly participants were healthy, non-smokers, and right-handed (confirmed using the Edinburgh Handedness Inventory). Participants were asked not to consume caffeine and alcohol-containing products 12 h before the experiment, and were told to come another time if they were ill or did not sleep well the night before the experiment. Written informed consent was obtained from all participants after explaining to them all the details of the experimental procedures as well as potential discomforts and risks. The studies were approved by the local Ethics Committee , performed in accordance with the Declaration of Helsinki.Thirty healthy participants took part in this study: 15 young  were performed and the partial eta squared (r) were used to identify relationships between variables. Statistical analyses were performed using IBM SPSS Statistics software .The data were tested for normal distribution using the Kolmogorov–Smirnov test, and all data were found to be normally distributed. A two-way mixed analysis of variance (ANOVA) with age as a between-group factor and with time as a within-group factor was taken. If significant effects were found, Sidak’s F = 3.43; p = 0.003; OP = 0.95; Figure 2). However the effect of age was not significant  = 0.26; p = 0.619; OP = 0.076; F = 0.66; p = 0.708; OP = 0.27; F = 3.34; p = 0.094; OP = 0.38; F = 0.64; p = 0.719; OP = 0.26; p > 0.05) in all cases interaction effect of task and age was not significant (p > 0.05).There was a significant increase in the number of “Incorrect NoGo” errors in the young men and the elderly men during the “Go/NoGo” task  = 0.71; p = 0.667; OP = 0.29; F = 6.77; p = 0.024; OP = 0.69; Figure 3). The RT of “Incorrect NoGo” significantly depended on the task  = 2.47; p = 0.02; OP = 0.85; F = 2.98; p = 0.012; OP = 0.55; p < 0.05). The after “Correct NoGo” RT of “Correct Go” did not significantly depend on the task  = 1.57; p = 0.154; OP = 0.52; F = 9.78; p = 0.009; OP = 0.82; p > 0.05). We found that the ratio of RT of “Correct Go” “after correct NoGo” to “Correct Go” did not significantly depend on the task  = 6.71; p = 0.137; OP = 0.49; F = 4.97; p = 0.041; OP = 0.69; p > 0.05).The RT of “Correct Go” did not significantly depend on the task  = 3.13; p = 0.005; OP = 0.93;F = 0.78; p = 0.39; OP = 0.12; Figure 4). Only in young men CV of RT of “Incorrect NoGo” significantly increased during PCL (p < 0.01). We established that there was not significant effect of task and effect of age on CV of RT both “Correct Go” and “Correct Go” after “Correct NoGo” (p > 0.05). In all cases the interaction effect of age and task was not significant (p > 0.05).The effect of task on CV of RT of “Incorrect NoGo” was significant (p < 0.05) in young men, whereas intrinsic motivation remained unchanged in elderly men . The prolonged performance of the task did not affect the success and overall motivation in young men  and elderly men . There were no significant differences in success, intrinsic, and overall motivation variables between young men and elderly men.The performance of the 2 h “Go/NoGo” task significantly decreased the intrinsic motivation from 23.2 (5.1) to 20.2 (5.8) (Table 1). Subjective fatigue, tension, and confusion significantly (p < 0.05) increased, and vigor decreased after the “Go/NoGo” task in the young men. Subjective fatigue significantly (p < 0.05) increased after the “Go/NoGo” task in the elderly men.There were no significant differences in mood variables before the “Go/NoGo” task between the young men and the elderly men (p < 0.05), while the accuracy of ST increased and the accuracy of CSDT decreased in the elderly men after the 2 h “Go/NoGo” task (p < 0.05) (Table 2). The RT variable of all cognitive tasks was significantly greater in the elderly men . The accuracy of ST, MT, and CSDT was greater, and the distance from the target of the Pursuit Tracking Task (PTT) was less in young men than in elderly man (p < 0.05) before and after the 2 h “Go/NoGo” task.In the young men, the RT and error rate of Memory Search Task (MST) significantly increased (p = 0.01) . The 2 h “Go/NoGo” task significantly decreased the switching/rest ratio of oxygenated hemoglobin for the young and the elderly men (p < 0.05). This ratio significantly more decreased for elderly compared to young men (p < 0.01). The ratio of oxygenated hemoglobin during ST after 2 h “Go/NoGo” to ST before the “Go/NoGo” task was 144.4 ± 4.2% in the young men and 33.5 ± 35.7% in the elderly men, respectively .The switching/rest ratio of oxygenated hemoglobin before “Go/NoGo” task in the elderly men was significantly greater than in the young men (p < 0.05); latency was significantly greater in the elderly men before the “Go/NoGo” task (Table 3). The “Go/NoGo” task did not affect the MEP amplitude (p > 0.05) there was no difference between the MEP amplitudes of the young and the elderly men (p > 0.05).Latency increased significantly after the “Go/NoGo” task in both groups of young and elderly men (p = 0.004); elderly men, 44.3 (10.6) kg and 44.9 (10.3) kg (p = 0.031). There were significant differences before and after the 2 h “Go/NoGo” task between young men and elderly men (p < 0.05).Muscle strength in the young men before and after “Go/NoGo” task was respectively 51.4 (11.4) kg and 47.6 (10.6) kg (p > 0.05) (Table 4).There were no significant differences in emotional intelligence items of four subscales  between the young men and the elderly men (p > 0.05); task-irrelevant score was 15.0 ± 5.6 for young men, and 12.0 ± 4.1 for elderly men (p > 0.05). Thus, there were no significant differences in thinking content during the “Go/NoGo” task between the young men and the elderly men (p > 0.05).According to the DSSQ score task-related score was 22.5 ± 5.3 for young men, and 21.6 ± 4.0 for elderly men (p < 0.05) in the young and the elderly men during the “Go/NoGo” task; mental demand increased significantly only in the young men during “Go/NoGo” task. However, temporal demand and load effort were significantly (p < 0.05) greater in the young men than in the elderly men during the “Go/NoGo” task (Table 5).Physical demand increased significantly , and directly for the elderly men ; the average number of “Incorrect NoGo” errors was respectively correlated . The increase in prefrontal cortex activity after exercise during the ST, significantly correlated (p < 0.05) with the correct cognition task performance after the “Go/NoGo” task, directly for the young men  and inversely  for the elderly men. In addition, the internal motivation significantly correlated with the overall increase in the number of errors  for the young men. The percentage increase in the total number of errors for the young men correlated strongly with tension  and vigor  before exercise.The average number of “Incorrect Go” errors significantly correlated with an increase in prefrontal cortical activation during ST before the “Go/NoGo” task, inversely for the young men  and neuromuscular function , cognitive function , and psychological variables  during and after a PCL (2 h “Go/NoGo” task).p < 0.05). Thus, contrary to our expectations, men young men showed more signs of cognitive fatigue than the elderly.The first finding of our study is that during the 2 h “Go/NoGo” task, both the young men and the elderly men had a significantly similar increase in the number of “Incorrect NoGo” (inhibition) errors (the effect of age was not significant), but the number of “Incorrect Go” errors was unchanged. The RT of “Correct Go” did not significantly depend on the PCL task, but it significantly depended on age (for the elderly it was longer). Only in young RT of “Incorrect NoGo” as well as CV of RT “Incorrect NoGo” significantly increased during PCL . The second main finding of our study was that despite the motivation level at the beginning of exercise not differing between the young men and the elderly men, the young men felt more fatigue, lack of energy, and mental and temporal demand during exercise, and their intrinsic motivation was more decreased during exercise. Taken together, psychological strain in the PCL was higher for the young men than for the elderly men. This is consistent with a study by Task performance strategy depends not only on the task prediction, but also on the current situation . For exaWhen people become tired, it is more likely that they continue to perform a task using automatic control . As our Our third main finding was that although this was unexpected, the elderly men performed some cognitive tests better (by about 10%) after PCL; they made fewer mistakes in the ST and CSIT. In sum, because many cognitive functions did not deteriorate in either the young men or the elderly men, we cannot claim that there is a significant difference between age groups. In our opinion, after “the task and motivation switch,” most cognitive tasks “recover.” This is consistent with data from other studies that manifestation of cognitive fatigue is specific, i.e., it depends not only on fatigue, but also on the specificity of tests establishing mental working capacity . BotviniThe fourth finding of our study was that prefrontal cortex activity increased during the ST more in the elderly men than in the young men before the “Go/NoGo” task. However, after the “Go/NoGo” task, the prefrontal cortex activity of the elderly men decreased, while in the young men it increased during the ST. The average number of “Incorrect NoGo” errors during 2 h “Go/NoGo” task significantly and directly correlated with an increase in prefrontal cortical activation during ST before the “Go/NoGo” task both in young and elderly man. However, the more the prefrontal activity increased during the ST after exercise, the greater the number of inhibition errors made by the elderly men, and the fewer the number made by the young men.Greater activation of the left dorsolateral prefrontal and superior parietal cortex during working memory was more evident in middle-aged than in young adults regardless of working memory load or fatigue condition . This coWe found that young men showed greater signs of cognitive fatigue than elderly men during a PCL, young men felt more fatigue after exercise than elderly men, and elderly men performed some cognitive tests better after PCL than young men. Because of the greater mental load and (possibly) greater recruitment (mobilization) of prefrontal cortex during a 2 h “Go/NoGo” task (PCL), there was greater mental and neuromuscular performance fatigue in young compared with elderly men. Prolonged task performance decreased the switching/rest ratio of oxygenated hemoglobin for the young and the elderly men; however, greater decrease was observed for elderly than young men. Finally, baseline prefrontal cortex activity during the switching task predicted mental performance changes during demanding mental load as the more highly the prefrontal cortex was activated, the better was the inhibitory control observed in young men, and the poorer was the inhibitory control observed in elderly men.All procedures performed in studies involving human participants were in accordance with the ethical standards of the local Ethics Committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.Written informed consent was obtained from all participants included in the study.The raw data supporting the conclusions of this manuscript will be made available by the authors, without undue reservation, to any qualified researcher.AS: conception and design of the study, acquisition, analysis, and interpretation of the data, and drafting the study. AT, EM, KV, DM, DK, DV, RS, and AE: collection, analysis, and interpretation of the data. AS and SK: drafting the study and revising it critically.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.


Meanwhile, the adsorption factors, adsorption characteristics, and mechanisms of GTF were investigated in batch adsorption experiments.Water stable Zr-metal–organic framework nanoparticles  have been solvothermally synthesized. PCN-224 NPs show spherical shape with smooth surface and particle size of approximately 200 nm. PCN-224 NPs can be stable in acid and aqueous solutions, as confirmed by powder X-ray diffraction. Gatifloxacin (GTF) adsorption measurements showed that PCN-224 NPs exhibit a high adsorption capacity of 876 mg·g Antibiotics are produced by micro-organisms and higher animals and plants throughout their life cycle. Antibiotics can resist pathogens or other types of secondary metabolites, can kill bacteria, and exert good inhibitory and killing effects on mold, mycoplasma, chlamydia, and other pathogenic microorganisms. Therefore, antibiotics are widely used in human medicine, animal medicine, biological science research, agriculture, animal husbandry, and the food industry. However, with the rapid development of human technology, the excessive use of antibiotics has become increasingly serious, and the amount of antibiotics in the ecological environment, especially in the water environment, is continuously increasing .Numerous studies have reported the presence of various antibiotics in waste water ,3, grounSeveral methods, including physical , biologiAs a new type of porous materials, metal–organic frameworks (MOFs) have attracted increasing attention because of their permanent porosity, high surface area, and the tunable chemical and physical properties of their pores. These properties make MOFs suitable for a variety of applications, in particular for various applications related to adsorption, such as solid phase microextraction , solid pIn MOFs, Zr-MOFs nanoparticles (NPs) have attracted increasing attention due to their specific electronic and optical properties, permanent porosity, high surface area, and easily tailorable structures and functions. Zr-MOFs can be used as adsorbents in the removal of nitrofurazone and nitrofurantoin from water . HoweverIn this paper, Zr-MOFs  NPs were synthesized  and usedN,N-dimethylformamide (DMF), acetone, pyrrole, propionic acid, benzoic acid, tetrahydrofuran, CH3OH, KOH, and ZrOCl2·8H2O were obtained from commercial sources , Energy Chemical , TCI , and J & K Scientific  and were used without further purification unless otherwise mentioned. Tetrakis(4-carboxyphenyl)porphyrin (H2TCPP) ligand was synthesized according to the literature [GTF was obtained from Sigma Aldrich Chemical Co. . Methyl 4-formylbenzoate, terature .1H-NMR spectra were recorded on a 400 MHz Varian INOVA spectrometer  and referenced to the residual solvent peak. Powder X-ray diffraction measurements were carried out on an analytical X-Pert pro diffractometer  with Cu-Kα radiation (γ = 1.5478 Å). Thermogravimetric analysis (TGA) was performed on a Mettler Toledo TGA  under N2 flow and heated from room temperature to 900 °C at 10 °C/min. Elemental analyses  were obtained on a PerkinElmer 240 elemental analyzer . Infrared (IR) spectroscopy spectra were collected on a Nicolet 330 FTIR Spectrometer  within the 4000–400 cm−1 region. Morphology was obtained using scanning electron microscopy (SEM) . In addition, the concentrations of GTF were measured using a UV–Vis spectrophotometer . Gas adsorption–desorption measurements of N2 (99.995%) on PCN-224 NPs was collected on the Micromeritics ASAP 2020 surface area and pore size analyzer . The as-synthesized crystals of PCN-224 were exchanged twice with dry acetone. The acetone-exchanged sample was degassed at 393 K for 12 h until the exhaust gas rate was 5 mm Hg·min−1 to produce the activated phases of PCN-224 for gas sorption measurements.2·8H2O and 1500 mg of benzoic acid were ultrasonically dissolved in 50 mL of DMF in a round-bottomed flask. Thereafter, 50 mg H2TCPP was added into the system, and the solution changed from colorless to dark purple. The mixture was heated in 120 °C for 1 h. After being cooling down to room temperature, dark brown powders were collected by centrifugation and washed for thrice with DMF and acetone . FT-IR : 3440(m), 2958(w), 1708(w), 1612(s), 1545(s), 1411(s), 1314(s), 1190(m), 1015(s), 866(w), 816(w), 779(m), 722(s); The analytical calculated elemental contents (%) for PCN-224 NPs are C, 44.04%; H, 2.62%; and N, 4.28%, while the founded results are: C, 45.49%; H, 3.02%; N, 4.07%.A total of 150 mg ZrOCl−1 standard stock solution of GTF was prepared by dissolving 0.4 g antibiotic in distilled water and diluting to a 500 mL brown volumetric flask. The standard stock solutions will be diluted to the required concentration in the experiment and stored in the dark below 4 °C.An 800 mg·L−1 GTF solution and 2 mg PCN-224 powder were introduced into 20 mL brown vials and mixed thoroughly by an ultrasonic cleaner for several seconds. Then, the vial was placed in a water bath oscillator with a shaking speed of 280 rpm at 25 °C for adsorption. After shaking for 240 min, the adsorption equilibrium was reached, and the suspension was centrifuged using a 0.22 μm. Then, the filtrate was collected, diluted, and analyzed immediately by UV–Vis spectrophotometry at 287 nm. Adsorption experiments were carried out under the same conditions. To evaluate the influence of pH, we adjusted the pH of GTF solutions from 3 to 9.5 by using 0.02 mol·L−1 NaOH and HCl. When we studied the effect of ionic strength, different amounts of NaCl particles were dissolved in the GTF solution, and the ionic strength were 0–2.5 mol·L−1. To study the effect of contact time, the contact time was in the range of 5–900 min. Adsorption isotherm and thermodynamic studies were carried out with initial GTF concentrations of 100–400 mg·L−1 at controlled temperature levels of 298, 308, and 318 K.A total of 10 mL of 300 mg·LThe crystallographic structure and phase purity of PCN-224 NPs were examined by powder X-ray diffraction c. No add2 gas sorption curves of PCN-224 NPs at 77 K were recorded to check for porosity , qt (mg·g−1), and the removal percentage was calculated using Equations (1) to (3), as follows:C0 is the initial GTF concentration (mg·L−1), Ce is the residual GTF concentration at equilibrium (mg·L−1), V is the volume of solution (L), and m is the mass of dry PCN-224 NPs (g).The performance of adsorbent is usually depicted by adsorption capacity and removal percentage. The adsorption capacity, −1) of GTF solution. This result indicated that the prepared PCN-224 NPs may offer excellent adsorption potency for the removal of GTF from aqueous solutions.In the pre-experiment, PCN-224 NPs showed excellent adsorption performance for GTF. All GTF (100%) was removed within 15 min when 10 mg PCN-224 powder was dispersed in 10 mL  and qt (mg·g−1) are the amounts of GTF absorbed on the sorbent in equilibrium and at time t (min), respectively. k1 (min−1) and k2 (g·mg−1·min−1) are the rate constants of the first-order and second-order adsorptions, respectively, and kd (g·mg−1·min−1/2) is the rate constant of intraparticle diffusion. I is a parameter related to the thickness of the boundary layer: increasingly high I means that large boundary layer effect. kf(h − 1) is the rate constant of liquid film diffusion, and A is the liquid film diffusion constant. qe,cal) of the pseudo-second-order model of 877.1929 mg·g−1 closely approximates the experimental value , that is, 876.7694 mg·g−1. These results indicate that chemical sorption is relatively dominant and controls the adsorption. The piece-wise linear regressions observed in the intraparticle diffusion plot (R2) value to fit the experimental data, proving that the rate-limiting step is mainly the molecular diffusion of GTF in the PCN-224 NPs.To study the relationship between the structure of adsorbent and adsorption performance, we used the data in the adsorption equilibrium time experiment to fit the kinetic model. Pseudo-first-order, pseudo-second-order, intraparticle diffusion kinetic, and liquid film diffusion models were used to describe the adsorption process. The four linear forms can be expressed as follows:ion plot c indication plot d yields The equilibrium isotherms at various temperature levels were studied by varying the initial concentration of GTF. Two well-known isotherm models, namely the Langmuir and Freundlich models, were employed in this study to analyze the experimental data.The equation linear form of the Langmuir model is as follows:qm is the Langmuir monolayer adsorption capacity (mg·g−1), qe is the amount of GTF adsorption at equilibrium (mg·g−1), Ce is the liquid-phase concentration of GTF (mg·L−1) at equilibrium, and b is the Langmuir constant (L·g−1) related to adsorption energy. KF is the Freundlich constant (mg·g−1 (mg·L−1)n−1/), and 1/n is the heterogeneity factor describing the adsorption intensity.The equation linear form of the Freundlich model is as follows:R2 values, the Freundlich model is more suitable than the Langmuir model for describing adsorption. The maximum adsorption capacity (qm) was decreased with increasing temperature, indicating that lower temperature facilitates GTF adsorption onto PCN-224 NPs and that the adsorption is an exothermic process. The 1/n values for the three temperatures studied were in the range of 0–1, which illustrated a favorable adsorption process.Sθ), enthalpy (∆Hθ), and the Gibbs free energy (∆Gθ), which are calculated using the following equations:qe is the amount of antibiotic absorbed on the sorbent at equilibrium, Ce is the solution concentration at equilibrium, Kc is the distribution coefficient, R is the molar gas constant (8.314 J·mol−1·K−1), and T is the temperature (K). The values of ∆Sθ and ∆Hθ can be calculated from the intercept and the slope of the linear plots shown in Hθ value was −16.609 KJ·mol−1, implying that adsorption is an exothermic reaction and that lower temperature is beneficial for absorbing GTF. The ∆Gθ and ∆Sθ values were negative, indicating that the adsorption of GTF onto PCN-224 NPs may be a spontaneous, physical sorption process.The thermodynamic parameters reflect the feasibility and spontaneous nature of the adsorption. The thermodynamic feasibility for GTF adsorption onto PCN-224 NPs has been demonstrated by evaluating the thermodynamic parameters, including entropy (∆R2 = 0.999). The adsorption isotherm was more consistent with the Freundlich model than the Langmuir model, indicating that the adsorption of GTF on PCN-224 NPs involves multilayer adsorption. The adsorption capacity of GTF on PCN-224 NPs was negatively related to the temperature. Moreover, the adsorption process was spontaneous and exothermic.In summary, Zr-MOF PCN-224 NPs were prepared and their adsorption behaviors were characterized and studied. Good removal capability for GTF from aqueous solution was obtained by the prepared PCN-224 NPs. The effect of contact time, pH, and ionic strength were considered and optimized. The adsorption kinetics, thermodynamics, and isotherms of GTF were calculated to characterize the adsorption behavior of GTF on PCN-224 NPs. The results exhibited high GTF adsorption and removal rates of PCN-224 NPs, and the pseudo-second-order model for adsorption kinetics was an excellent fit (


Videogame addiction has been suggested as a tentative disorder in 2013 by the American Psychiatric Association (APA) and was recently officially recognized as a mental health disorder by the World Health Organization (WHO). Although a few studies have identified attention deficit and hyperactivity disorder (ADHD) as a key risk factor for Internet Gaming Disorder (IGD), the interplay between ADHD and IGD symptoms with gender differences across cultures remains to be further examined.This study examined the moderating effects of gender in the association between ADHD and IGD across two nations.age = 23.01, SD = 3.35, Minage = 18, Maxage = 31, Males n = 121, 73.80%) and 457 U.S.-North American  Massively Multiplayer Online (MMO) players aged between 18 and 29 years.A cross-sectional online survey was developed to recruit 164 Australian  and Attention Deficit Hyperactivity Disorder (ADHD) have been associated.•A combined sample of Australian and American Gamers was examined.•The moderating effects of gender and country of origin were assessed regarding the ADHD-IGD associations.•American Males gamers presenting with ADHD were at higher IGD risk. In partAlongside the increased popularity of online gaming, research examining its potential effects has grown exponentially . Indeed,bona fide addiction negatively affecting a minority of gamers  , as a glIn this context, variations considering IGD vulnerability have been illustrated involving specific developmental stages  and game genres , as well as individual  and contextual characteristics  have been highlighted as potential risk factors for IGD . Accordi1.3Several behavioral problems, risky behaviors and psychiatric disorders have been consistently associated with IGD symptoms in both cross-sectional and longitudinal studies . Some ofIn that context IGD has been viewed as both a disorder that may be secondary to ADHD or one with a reciprocal relationship with ADHD on the basis of reward seeking gratification  and IGD will vary by gender.4HThe IGD related effect of the interplay between genders with different types of ADHD symptoms could vary between USA and Australian gamers.This study conceptualized IGD behaviors upon a continuum from absence of behaviors to predominance  to exploThis exploratory hypothesis is supported by vertical individualism variations, which could interfere with the significance of achievement gaming-drives .Finally, the potential prevalence of gaming disorder, as defined by scoring above the recommended cut-off of the Internet Gaming Disorder Scale-Short-Form  and 457 U.S.-North American . The estimated maximum sampling error regarding the 163 Australian gamers is 7.68%, for the 457 US gamers is 4.58% and for the 620 participants of the combined sample is 3.94% .The sample consisted of 163 Australian  and internet use (i.e. daily time of internet and gaming use on week days and the weekend) information before completing a battery of self-report questionnaires.2.2.12.2.2. Adult ADHD Self-Report Scale (ASRS-v1.1)The severity of IGD symptoms was measured using the IGDS9-SF, which reflects the nine IGD criteria . The scahow often do you make careless mistakes when you have to work on a boring or difficult project?) and nine items assessing hyperactivity-impulsivity (i.e. how often do you fidget or squirm with your hands or your feet when you have to sit down for a long time?). All items are answered to 5-point Likert scale (1 = “Never” to 5 = “Very Often”), and total scores for the inattention (ranging between 9 and 45) and the hyperactivity-impulsivity (ranging between 9 and 45) subscales are produced by accumulating the relevant item points. The full ADHD scale score is produced based on the addition of the scores of the two subscales resulting a total score ranging from 18 to 90 points, with higher scores indicating higher ADHD symptoms. The internal reliability scores of the two subscales and the full scale were satisfactory for both samples .Adult ADHD symptoms were measured using the ASRS-v1.1 . The ins2.3SurveyMonkey  and/or Amazon Mechanical Turk . Participation was voluntary and participants were provided with a Plain Language Information Statement (PLIS) to consent before they engaged. They were free to withdraw at any time prior to submission of the survey. Online collection was preferred, as it provides higher accessibility and representation of internet gamers.The study was approved by the ethics committees of the Palo Alto University (USA) and Federation University . Eligible participants were MMO gamers, residents of Australia and the USA, aged between 18 and 29 years. The study was advertised online and data was collected via 2.4To assess the association between inattention and IGD behaviors , two on, these wSimilarly, to assess the association between hyperactivity-impulsivity and IGD behaviors , two mor, with the only difference of using inattention symptoms as the independent variable. This aimed to assess whether there was a moderating effect of gender on the association between inattention and IGD and whether this differed between countries.To assess 3N = 18) compared to the Australian . The USA sample also reported more ADHD symptoms compared to the Australian sample (44 ± 15.4 vs. 29.7 ± 10.7) for both inattention and hyperactivity-impulsivity., the slope of the regression line was statistically significant  = 122.69, p < .001). Specifically, 22.9% of the IGD variance was explained by inattentive ADHD (R2 = 0.229). For each point of increase in inattention, IGD increased by 0.44 . The Australian sample results showed similar results, with a statistically significant regression slope  = 13.36, p = .001). Specifically, 7.6% of the IGD variance was explained by inattention (R2 = 0.076). For each point of increase in inattention, IGD scores increased by 0.45 . To assess the potentially moderating effect of country on the association between Inattention and IGD behaviors the following model was additionally estimated:Considering b2 = 1.05, t = 0.44, p = .659, LLCI = −3.60 – ULCI = 5.69] and the interplay between inattention and country  on IGD were not statistically significant, with the latter explaining <0.01% of the IGD score variance (R2 = 0.000).Findings indicated that country  on IGD was insignificant explaining <1% of the IGD variance (R2 = 0.004).The interaction between hyperactivity-impulsivity and country [To address F = 25.00, p < .001; see b8 = 7.83, t = 2.27, p = .024). Similarly, an inspection of the three-way interaction coefficient between hyperactivity-impulsivity, country and gender indicated a significant protective interaction considering IGD . Specifically, when an MMO gamer presented concurrently with a higher point (than the intercept) in hyperactivity-impulsivity, was male and Australian, then IGD reduced by 0.68  = 18.89, p < .001; see b = − 0.59, t = −2.00, p = .046). Specifically, when an MMO gamer presented concurrently with a higher point (than the intercept) in inattention, was male and Australian, then IGD reduced by −0.59 . Specifically, more hyperactive-impulsive, as well as inattentive males in the USA presented higher on IGD behaviors than their Australian counterparts. These results reinforced past literature hypotheses considering culturally related fluctuations of the effects of IGD risk factors. Furthermore, findings highlighted that such variations need to be taken into account when studying individual IGD risk effects, such as gender and the reported levels of other concurrent psychopathology. Finally, our data indicated that nearly 4% of USA gamers and nearly 2% of Australian gamers are potentially at risk for disordered gaming. While these proportions are lower than that previously reported , they do4.1The study envisaged and confirmed that MMO gamers experiencing higher inattention symptoms would report elevated IGD behaviors , and thi4.2Similarly, the hypothesis  that MMOdevelopment of IGD, longitudinal studies must be conducted to answer important empirical questions about the ADHD-IGD temporal association.Overall, the present study's results serves to increase the current knowledge surrounding the association between hyperactivity-impulsivity and IGD symptoms in two significant ways. First, the present study differentiates the effect of hyperactivity-impulsivity from inattention symptoms on IGD behaviors. Second, as with the case of inattention related IGD effects, it indicated that this effect exhibited cross-cultural resilience across groups of Australian and USA gamers. Nevertheless, as mentioned earlier studies across more diverse cultures are needed. Furthermore, in order to assess 4.3The present study's hypotheses , H4 thatIn that line, the variations of the hyperactivity-impulsivity, as well as inattention symptoms and IGD behaviors associations revealed here, consistently to the detriment of USA gamers appear to also align with past literature . Culture4.4The importance of these results can be reflected in their ability to identify the differential impact that the interplay between gender and culture may introduce on the associations between the two different types of ADHD symptoms and IGD. Specifically, emphasis on the higher IGD risk of more vertically individualistic male players, presenting with either inattention or/and hyperactivity-impulsivity symptoms should be taken into consideration when planning relevant prevention and treatment interventions worldwide.Nevertheless, the present study has significant limitations. Among others, these include the use of self-report questionnaires, where participants' ability to accurately estimate their gaming time is questionable see . AlongsiThe authors report no conflicts of interest.


Helicobacter pylori has been associated with gastric mucosal damage and gastroduodenal diseases, studies evaluating gastric cancer patients are scarce. We investigated whether the functional oipA “on” status was associated with gastric cancer in the North-eastern Brazil, region with high prevalence of gastric cancer.Although, outer membrane protein OipA of H. pylori positive subjects . oipA was assayed by polymerase chain reaction (PCR) and DNA sequencing. cagA and vacA status were evaluated by PCR.We included samples from 95 H. pylori strains had functional oipA. In adults, the oipA “on” status  and increasing age  were independently associated with gastric cancer in a logistic model. The oipA “on” status  was also associated with first-degree relatives of gastric cancer patients when compared with gastritis. The frequency of oipA “on” status did not differ between children and adults (P = 0.87). The oipA “on” status was significantly correlated with the presence of cagA and vacA s1 m1.Overall 81.1% of the oipA “on” status was independently associated with gastric cancer and first-degree relatives of gastric cancer patients in North-eastern Brazil. Gastric cancer is the fifth most common cancer and the third leading cause of mortality among men and the fifth among women, with more than 700,000 deaths per year worldwide . The malHelicobacter pylori is a well-recognized bacterium that chronically infects the stomach of approximately half of the world’s population being more prevalent in developing countries. Chronic H. pylori infection is considered the strongest risk factor for distal gastric adenocarcinoma  and 9 CT [3.9% (3/77)] repeat patterns. Among the oipA “off” status (n = 18), the following CT repeat patterns were found: 5 CTs (n = 4/22.2%), 7 CT (n = 11/61.1%), 8 CT (n = 2/11.1%), 9 CT (n = 1/5.6%). The CT pattern frequency according to the different H. pylori positive subjects are shown in Table The number of CT repeat patterns ranged from five to nine Table . The 6 CTs n = 4/2.2%, 7 CoipA “on” status was found in 68.8% (11/16) of the H. pylori strains, without significant difference from that of the adults with gastritis .The functional “on” status was observed in 91.7% (22/24) of the adult patients with gastric cancer, in 60.9% (14/23) of the patients with gastritis and in 93.8% (30/32) of the first-degree relatives of gastric cancer patients .No significant difference was observed in the frequency of oipA “on” status was positively correlated with the presence of cagA  and vacA s1 m1 , but negatively correlated with the presence of vacA m2 allele . In the gastric cancer patients, the percentage of simultaneously positive oipA “on” status and cagA was 91.7% . High positive correlation was also observed between in frame oipA and s1 m1 genotype . In respect to the gastric cancer relatives, high percentages were also observed in oipA “on” status and cagA positivity (88.0%) as well as in oipA “on” status and s1 m1 vacA genotype (83.3%).In the gastritis group, the P = 0.08 – Mantel Haensel two-tailed chi-square test). In the group of gastritis patients, the pattern of the antral gastritis was more frequently mild than moderate/severe (4/9–44.4% vs. 5/19–26.3%). Otherwise, because the frequency of oipA “on” status was very high in the group of the first-degree relatives of gastric cancer patients, no association was observed between the presence of functional oipA and the degree of mononuclear and polymorphonuclear cells in the gastric mucosa.The presence of gastritis in the antral and oxyntic mucosa was higher in the first-degree relatives of gastric cancer patients (11–52.4%) than in the gastritis patients (3–13.0%), , similarly to that observed in Bulgaria (81.0%) [oipA “on” status is lower than that observed in the current study. In children, the frequency of oipA “on” status was higher in the H. pylori strains from Brazil than in those from Portugal (49.6%) [oipA “on” status observed in the children is in agreement with high prevalence of other H. pylori risk factors we have previously observed in children from the same population [Gastric cancer remains a major healthy problem, especially in developing countries. Unfavorable outcome of cytokine  and in mcytokine . In both (81.0%) , Colombi (81.0%)  and Vene (81.0%) . It has  (81.0%)  and Mala (81.0%) . Otherwi (81.0%)  and Nort (81.0%)  demonstr (49.6%)  and USA  (49.6%) . This hipulation . Taking oipA status was significantly associated with gastric cancer, even after adjusting for confounding factors as reported in Colombia [oipA “on” status was significantly more frequent in the first-degree relatives of gastric cancer patients than in those with gastritis. This finding is a novelty because we are not aware of studies evaluating oipA H. pylori virulence marker in such individuals who are at increased risk of gastric cancer that is determined by both bacterial and host factors. It has to be emphasized that first-degree relatives of gastric cancer patients are prone to be colonized by bacteria that circulate within a family. In fact, genetic fingerprint methods have demonstrated genetic homogeneity in the H. pylori strains within a family. In a previous study, we have shown that first-degree relatives of gastric cancer patients living in the same Brazilian region were colonized with cagA-positive and vacA more virulent H. pylori strains with the same characteristics of those isolated from their parents/siblings with gastric cancer [Remarkably, we observed that the functional Colombia , where tColombia . Furtherc cancer .oipA “on” as a gastric cancer risk factor include the ability of the bacterium carrying a functional oipA to attach to the gastric epithelial cells [Evidences that point to the al cells , 34, to al cells , 34, apoal cells .H. pylori gastric colonization induces epithelial cells to produce a series of pro-inflammatory and anti-inflammatory cytokines and chemokines; among them IL-8, a potent neutrophil chemotactic and activating peptide [oipA functional gene is associated with increased IL-8 production by gastric cancer cell line [ peptide , 11, 36.ell line , 37 and oipA sequencing patterns we detected, the 6 CT repeat pattern was the most common oipA “on” status found, in consonance with other studies in Western countries [H. pylori strains had less than 5 CT repeats in the oipA gene [Among the seven ountries , 12, 24,ipA gene .oipA “on” status could not be discriminated from the other H. pylori virulent factors because all of them were highly frequent and closely linked to each other, especially the in frame oipA and cagA virulence factors that have been considered as having a synergistic effect on the pathogenesis induced by H. pylori.In this study, the oipA “on” status between gastric cancer/first-degree relatives of gastric cancer patients and patients with gastritis alone, limitations of our study should be considered. An eventual bias is the small sample size that may preclude associations between oipA “on” status and age and gender. In addition, the small sample size impaired us to evaluate associations between the expression of oipA “on” status and scores of gastritis in the group of first-degree relatives of gastric cancer patients. Because, we evaluated H. pylori strains of people living in one of the least developed Brazilian regions, it is possible that these findings differ among the different regions of our country that have marked difference in the prevalence of H. pylori-positive status as well as in the prevalence of gastric cancer, which points to the need for further studies on this subject in other geographical regions.Although the sample size had been enough to discriminate the results of high frequency of oipA “on” status is associated with gastric cancer and with first-degree relatives of gastric cancer patients in North-eastern Brazilian population.In conclusion, the