When voters in Washington State banned hound hunting with an eye to protecting cougars, they unwittingly triggered a chain of events that wildlife biologists are still trying to understand.  Now the forests are removed, the land covered with fields of corn, orchards bending with fruit and the magnificent habitations of rational and civilized people.”—John Adams, 1756 [.Braced for savagery and sacrifice, European settlers in the New World came to the Pacific Northwest to tame the final frontier, the last refuge of “dismal wilderness.” While colonists in the East were poisoning, shooting, and trapping cougars to extinction during the 1880s, hundreds of thousands of pioneers flooded into what would soon become the new state of Washington. Following the well-worn pioneer playbook, Pacific Northwest immigrants converted forests to farmland and pasture and, fearing local predators as unacceptable threats to life, property, and game, paid bounty hunters to destroy all carnivores, large and small.It would take over 30 years to exterminate the wolf, and several more decades to nearly eliminate the cougar—whose famously reclusive, solitary nature may have helped the cat survive a systematic eradication effort. Yet even against a backdrop of ongoing persecution, complaints of cougar attacks on livestock and game continued apace, and legislators, assuming that more complaints meant more predators, increased incentives for hunters to thin the population. By 1940, two United States Fish and Wildlife Service senior biologists reported that cougars had been “exterminated in practically all of their former range in the United States and are fast being eradicated from many parts of the West” .Ironically, a measure passed to protect wildlife triggered a chain of events that led to the highest rates of human-caused cougar mortality since the height of the bounty era—even as the public clamored for higher cougar harvests.Puma concolor, about 400,000 years ago, went extinct in North America during the last ice age, 10,000–12,000 years ago, then recolonized the continent from surviving populations in Central and South America [P. concolor lost two-thirds of its historic range during the bounty era, which ended in most western states in the 1960s. Though cougar abundance increased in the West after states reclassified the cats as game animals, a designation that afforded them limited protection, the World Conservation Union considers P. concolor “near threatened” and warns that the species may soon qualify as “vulnerable” if current persecution and habitat degradation trends continue [Genetic evidence suggests that cougars evolved as a distinct species,  America , acquiriAlthough reliable population estimates are notoriously difficult to generate for shy, wide-ranging, low-density animals, the Washington Department of Fish and Wildlife (WDFW) estimates that 2,500 to 4,000 cougars now inhabit the state—and they're finding it harder and harder to steer clear of humans. Washington's human population increased 21% between 1990 and 2000, far faster than the national average of 13% .Worried about the loss of prime cougar habitat and persecution of a top predator, animal welfare and environmental groups sponsored a statewide initiative (I-655) to outlaw the use of hounds to hunt cougars, a longstanding rural tradition—and the most efficient method of killing cougars—regarded as cruel by many city-dwelling voters. Anxiety over cougars, always a hot-button issue in the sparsely populated counties in northeastern Washington, reached a fever pitch after I-655 passed in 1996.Yet in spite of predictions that an exploding cougar population would leave a trail of mutilated horses, dogs, and children, the measure's impacts were neither what supporters had hoped nor opponents feared. Ironically, it triggered a chain of events that led to the highest rates of human-caused cougar mortality since the height of the bounty era—even as the public clamored for higher cougar harvests. Wildlife biologists are still trying to understand the impacts of such heavy hunting on the ecology, behavior, and persistence of one of Earth's most secretive species. Whether they can find a way to help Washington residents and cougars coexist remains to be seen.Catherine Lambert originally set out in 2002 to study regional variations in the reproductive response of cougars in the Pacific Northwest, but shifted gears when her radio-collared research subjects kept turning up dead. Lambert, then a student at Washington State University's Large Carnivore Conservation Laboratory, was shocked to find that nearly half of 52 radio-collared cats had been shot by hunters or in response to livestock depredation attributed to cougars. The mounting body count suggested that the population could be in serious trouble from overhunting. But Lambert's field observations ran directly counter to the popular belief that I-655 had triggered a cougar population explosion.Contrary to popular belief—and the rationale behind legislation authorizing emergency and public safety hunts—increased complaints did not signal a growing cougar population.That was because WDFW officials, well aware that losing hound hunting could reduce the number of cats killed, had liberalized hunting regulations to maintain traditional harvest levels. The agency extended the hunting season by six months, doubled the legal bag limit, and rolled cougar “tags,” or licenses, into big-game packages, which made them far more attractive to elk and deer hunters, known as “boot hunters.” Before the ban, WDFW sold 1,000 cougar tags a year. The new policy gave tens of thousands of deer and elk hunters the legal right to shoot cougars.The Idaho Observer, bearing the horror movie headline “Cougar Carnage at the Promised Land Ranch” and featuring a grisly photo of a wounded colt [But perceptions die hard. Despite the agency's efforts—and even though I-655 allowed the agency to use hounds to protect the public—the incidence of cougar complaints, which averaged about 250 a year before I-655, more than doubled the following year before peaking at 936 in 2000. Cougar–human conflicts increased along with public anxiety, particularly in Okanogan County, an area where apprehension about cougars runs deep—the state's only recorded fatal attack on a human occurred here in 1924. The mood is captured by a 2003 column by Joel Kretz in a crusading libertarian monthly published across the border, ded colt .As frustration with the hound-hunting ban festered, Okanogan County commissioners threatened to defy state law by declaring open season on the “growing” cougar population, and by 2004, nine statewide bills had been introduced to reverse or circumvent I-655; two passed. Speaking for the Okanogan Farm Bureau, Kretz testified in favor of one that authorized hounds for public safety hunts and another that sanctioned emergency safety hunts in a pilot program that gave commissioners in five northeastern counties local control over cougar management. After years of complaining to politicians and the press about an “exploding” cougar population, Kretz was elected state representative from four of the five pilot counties in 2004. In 2007, he introduced a bill to extend the emergency safety hunt program another year.As the “cougar problem” was debated on editorial pages, in public forums, and at state and county hearings—and the management of an enigmatic species moved from the hands of wildlife biologists first to voters and then to politicians—58,000 deer and elk hunters hit the woods with cougar tags in their pockets. And Catherine Lambert, worried about losing so many collared cougars, set out to test her suspicion that public perception about an exploding cougar population was dead wrong.When Lambert joined the Large Carnivore Conservation Laboratory in 2002, its director, Rob Wielgus, was investigating declines of mule deer and endangered mountain caribou in the Selkirk Mountains, their last stand in the lower 48 states. Years of intensive timber harvest had transformed the ancient stands of old-growth forests rich with arboreal lichen, the mainstay of the caribou winter diet, into clear-cut blocks rife with forest edges and early seral vegetation like seedlings, saplings, and immature trees, destroying critical caribou habitat and forage.White-tailed deer—historically rare in these parts—thrive on the immature vegetation left behind by forestry practices, and their numbers rose as those of native prey species declined. One explanation for the white-tails' success could be that they outcompeted native species for resources. But Wielgus found support for an alternative hypothesis called apparent competition, a negative interaction between prey species that occurs due to shared enemies rather than shared resources. As the white-tails invaded native ungulate range, moving upland in the summer, cougars followed, and their numbers expanded along with their prey base—for a while. Cougar predation on white-tails was density-dependent—it increased or decreased in relation to population growth—but continued to increase on caribou and mule deer even as their populations declined ,9. This Without intervention—such as culling cougars or changing forestry practices to discourage white-tails—Wielgus feared that neither the Selkirk caribou nor the mule deer populations would recover. Wildlife managers in the region agreed to increase hunting to limit predation. But why were cougars selecting for the native species? At first, Wielgus wondered if mule deer had hybridized with white-tailed deer and were somehow easier to kill, but DNA analysis showed no white-tail gene introgression into mule deer taken by cougars. Then his team analyzed “a huge dataset” on deer kills from global positioning system readings and realized that male cougars were killing white-tails at lower elevations, while females were killing mule deer at higher elevations.Cougars appeared to be causing consternation everywhere, eating endangered caribou and deer and attacking livestock and pets, and even the occasional human. Cougar attacks on humans are extremely rare—lightning strikes are more common—but eight of nine documented attacks occurred in the 1990s, including two serious attacks on children in northeastern Washington, providing fodder for the post-I-655 legislative blitz to expand hunting. What if all the problems were the result of a hunted—rather than a growing—cougar population? Evidence from studies on African lions  and wolvWhen Lambert began work on Wielgus's cougar project, his team had already started to capture and radio-collar 52 cats, first in study sites around the Selkirk Mountains in northeastern Washington, southern British Columbia, and northern Idaho, and later in another site in Colville National Forest in northeastern Washington see . The tea“This is where it gets really depressing,” Wielgus told a recent national meeting of science writers, where he presented his latest results. Aside from some older females, “we don't have any four-year-old cougars left.” Hunters, as Lambert discovered firsthand, accounted for 92% of cougar deaths—and indirectly killed five of 21 dependent kittens by shooting their mothers.2 at the beginning of the study to 0.85 cougars/100 km2 in the last two years. As Lambert reported last year in the Journal of Wildlife Management [2 in Alberta, Canada, for example, and 3.5–3.7 cougars/100 km2 in British Columbia).Based on an annual census, the minimal total density fell from 1.46 cougars/100 kmnagement , even thThe population was growing at the start of the study, when Wielgus discovered cougar selection on caribou and mule deer, but started to decline by 30% a year in 2000—just when complaints reached an all-time high. If current harvest rates continued, the cougar population would disappear within 30 years. Contrary to popular belief—and the rationale behind legislation authorizing emergency and public safety hunts—increased complaints did not signal a growing cougar population. “As complaints were going up, the population was tanking,” Wielgus says.The intensive hunting in the Selkirks did achieve one thing: it relieved predation pressure on caribou and mule deer. Mule deer populations have recovered beyond expectation, but white-tailed deer are also increasing—at the rate of 30% a year. The strategy just facilitated the invasion of the white-tail, Wielgus says, which will likely outcompete mule deer for resources down the road.Wielgus's team continued studying the population in Colville National Forest, another area with fears about a growing cougar population and heavy hunting, though on a smaller scale. Based on low survival and maternity rates—kitten survival rates were also low—the population was in rapid decline, mostly due to female mortality. Yet by census count, the population appeared stable over time, but not sustainable. The team found more juveniles than expected, no decline in total or adult density, and a shift in population structure toward younger independent males. The hunted population acted as a sink, attracting immigrants and younger animals, which masked the loss of females. But males won't stick around if there aren't any females left. And a population without females has no future.Wielgus saw a similar dynamic with grizzly bears when he tested the notion that trophy hunting increases offspring production, survival, and population growth by reducing the abundance of competitive or cannibalistic adult males. Deer and other traditional game animals typically respond to predation (or hunting) with increased reproduction and survival. But top carnivores, which have not adapted to predation over evolutionary time, should not be expected to respond like prey species, Wielgus reasoned. Instead, he found that hunting older adult male grizzly bears in small populations attracted dispersing, potentially infanticidal males, led to increased sexual segregation and reduced reproduction, and ultimately compromised population growth and persistence. Reduced cub production, Wielgus argued, occurred because adult females moved into territories where resources and, presumably, infanticidal males were scarce .The same thing appears to be happening with cougars. “As we kill all these big resident adult animals, the younger guys come to the funeral,” Wielgus says. And that could explain cougar selection on mule deer. The immigrant males hang out in lower elevations, killing white-tailed deer where prey densities are very high, but females move to higher elevations—where prey densities are lower—and kill mule deer incidentally. It stands to reason that females would go where prey densities are high, but they don't. Wielgus is testing the possibility that females go to resource-poor areas to avoid immigrant males—which easily travel 150 miles (240 km) to find a potential mate—to protect their kittens. He doesn't have direct evidence of infanticide, but notes that more kittens turn up dead when unrelated immigrant males enter the system than when their fathers are there.Wielgus found the probable source population for the Colville immigrants in another study area to the southwest, where white-tailed deer are still rare. Unlike both heavily hunted populations, kitten and adult female survival rates were high—and adult male survival rates were twice as high. In this “lightly hunted” population, the mule deer population was healthy and cougar complaints were low. Hunting was acting as a form of habitat degradation. The lightly hunted populations have stable habitat use, home ranges, and population growth. But in heavily hunted populations, “we appear to have chaos,” Wielgus says, with no adult males, an influx of immigrants from surrounding areas, home ranges and densities “shifting all over the place,” more infanticide, and far more cougar–human conflicts. “And we suspect—this is what we're studying now—that these teenage males cause more problems than older residents and that this heavy harvest exacerbates the problem rather than making it better.”While state law prohibited WDFW officials from commenting publicly on I-655, agency biologists saw the two legislative measures authorizing hounds for safety hunts as an opportunity to reassert control of cougar management see . For theIronically, once the initiative to ban hound hunting passed, “presumably to protect cougars,” Rob Wielgus says, “it resulted in a big harvest of cougars, a decline in the female component, and the influx of teenage males. The road to hell is paved with good intentions. All my data suggest that we should go back to hound hunting, which is regulatory, density-dependent, and sustainable.”Whereas hound hunters typically select for trophy toms, deer and elk hunters kill indiscriminately. “Hunters are up in a tree stand waiting for a deer to come by and all of a sudden they see a cougar,” says WDFW's Rich Beausoleil. Before the big-game package was created, he says, “they wouldn't have had a license to take that cougar, but now, because they have the tag, they take it.”Beausoleil and Donny Martorello, a WDFW carnivore expert, studied harvest statistics before and after the 1996 ban . Before Wielgus thinks that hound hunting may lead to a self-regulated, density-dependent harvest, a theory he's testing now. Hound hunters typically go where residents have reported an incident, and tend not to overharvest, since cougars occur at such low densities. But deer and elk hunters kill far more cougars incidentally. “We documented that as cougar numbers go down, deer numbers go up, and what I think is happening is that when deer numbers go up, you get more boot hunters, which means more guys with cougar tags in their pocket, so they kill more cougars, which means more deer, which means they kill more cougars,” he explains. The smaller the cougar population, the higher the hunting. “That's inversely density-dependent,” Wielgus says. “Some people call that the road to extirpation.”For Wielgus, mounting evidence over the past decade argues that it's time to rethink wildlife management models. “We're learning all kinds of things that are counterintuitive,” he says, like the notion that shooting animals may not reduce their numbers. Traditional management models have been based on white-tail populations in Pennsylvania and deer mice, he says, but large carnivore behavior and population dynamics are completely different.As for understanding the dynamics of cougar predation on endangered prey, Wielgus says that it's important to ask why the predators are there in large numbers. “If there's a really high density of white-tails, the predators are going to come in, so the long-term solution is getting the habitat back in shape such that it's not so attractive to white-tailed deer,” he says. “And if white-tails are expanding into areas where they're historically nonnative and wolves aren't there to kill them, which they historically may have done elsewhere, something has to take their place.” Hunting may be the best short-term solution, Wielgus says. “You could say, ‘Let's not hunt the white-tails,’ but then it's bye-bye mountain caribou and bye-bye mule deer. Meanwhile our entire ecosystem is suffering.”Heavy hunting, as Wielgus showed, alleviated predation pressure on endangered ungulates only by sending the cougar population to the brink of collapse. And heavy hunting on a smaller scale didn't even reduce the population, or the complaints, because of increased immigration. Since predatory behavior is learned—a cougar might discover that caribou herds concentrated in small forest patches or livestock in fenced pastures make easy pickings—individual cats can cause a lot of trouble see . When WiWDFW is now seeking public comment on its next game-management plan , which wFor the wildlife officials who spend their days mediating conflicts with cougars, the prospects for coexistence depend on public education. “If you put your dog out to do his business at 1 a.m. with no lights on and no noise, and a cougar just happens to be passing by, it's likely to figure out that lunch comes out the back door every night at one in the morning,” says WDFW enforcement officer Jim Brown.Convincing the public to accept top predators as an integral part of a healthy landscape is Beausoleil's long-term goal. But it won't be easy. Gray wolves have been sighted around Lake Chelan, just west of Okanogan County, far from established packs in neighboring Idaho. “If the wolves get here, we won't even be thinking about cougars,” says State Representative Kretz. “They're a hundred times worse.”Hunting was acting as a form of habitat degradation. The lightly hunted populations have stable habitat use, home ranges, and population growth. But in heavily hunted populations, chaos prevails.When top predators like cougars and wolves disappear, surprising things happen. By creating a “landscape of fear,” predators change prey behavior. Reintroducing gray wolves into Yellowstone National Park drove deer, elk, and moose out of willow stands, releasing grazing pressure on songbird habitat and increasing songbird diversity . The absSuch benefits are a tough sell among those who view large carnivores as threats to life and property. While public safety is still a top priority for WDFW, agency biologists, unlike politicians, must also worry about the needs of resident wildlife. “One of the things we'll never get a handle on is the folks who move to the end of a box canyon in the middle of nowhere, and maybe they come from the city, and they see a cougar and say, ‘Hey, I saw a cougar, you've got to remove him,’ ” says Beausoleil. “Well, no, that's not what we do. You're living in cougar country now.” He hopes that one day the developers whose brochures tout all the bounding hills, wildflowers, deer, and elk will tell people about all the bear and cougar there too.In 1946, US Fish and Wildlife biologist Stanley Young wrote that cougars “are so destructive to man's interests that they cannot be tolerated except in the wildest areas” . But he The days when wildlife managers viewed the cat of many names as vermin to be eradicated are long gone. Modern managers promote predators' role as guardians of ecosystem integrity, but they are also employees of the state and must balance the needs of the species with the will of the electorate. As America's great stretches of wilderness rapidly disappear into the transfigured landscapes of advancing development, the fate of the cougar depends on whether “rational and civilized people” can see that the world would be a poorer place without predators.


We introduce a generalization of the well-known ARCH process, widely used for generating uncorrelated stochastic time series with long-term non-Gaussian distributions and long-lasting correlations in the (instantaneous) standard deviation exhibiting a clustering profile. Specifically, inspired by the fact that in a variety of systems impacting events are hardly forgot, we split the process into two different regimes: a first one for regular periods where the average volatility of the fluctuations within a certain period of time  For the last years the physical community has broaden its subject goals to matters that some decades ago were too distant from the classical topics of Physics. Despite being apparently at odds with the standard motivations of Physics, this new trend has given an invaluable contribution toward a more connected way of making Science, thus leading to a better understanding of the world surrounding us In this work, we make use of the celebrated heteroscedastic model, the instantaneous volatility for mere historical reasons. Traditionally, a Gaussian is assigned to the random variable viz., price fluctuations modelling, the case The Engle's formulation of an autoregressive conditional heteroscedastic  As described above, the statistical features of the macroscopic observables are the result of the nature of the interactions between the microscopic elements of the system and the relation between microscopic as well as the macroscopic observables. In the case of the “financial” i.e., impact) in the system, no matter when they took place, by changing the surrounding conditions as agent-based models suggested In our case, we want to emphasize the fact that people tend to recall periods of high volatility . Thence, our proposal is nothing but the use of simple mechanisms that in a coarse-grained way master a good part of our decisions.We are therefore dealing with a model characterized by 5 parameters, namely: In this section we present the results obtained by the numerical implementation of the model. For comparison, we will use the results of a prior model that can be enclosed in the class of We have adjusted the probability distributions of Concerning the persistence of the volatility, we have settled on the Detrended Fluctuation Analysis (DFA) Let us now present our results for As we increase the value of Concerning the instantaneous volatility, Cases for which the kurtosis excess is relevant  log-index fluctuations, ent  see . Compari see .It is worthy to be mentioned that all the three values of the parameters are plausible. First, within an application context, i.e., fat tails and strong persistence), the distributions are very well described by a type-2 Gumbel distribution in large part of the domain, which explains the emergence of the tails.We have studied a generalization of the well-known http://mesonpi.cat.cbpf.br/ssolar/).Our results have been obtained from numerical simulation using code written in fortran language and run on the 64-bit ssolarII cluster (


IL28B gene already found in different populations, mostly infected with viral genotypes 1 and 3. Egypt has the highest prevalence of HCV infection in the world, which is mostly due to viral genotype 4. We investigated the role of several IL28B SNPs in HCV spontaneous clearance in an Egyptian population. We selected nine SNPs within the IL28B genomic region covering the linkage disequilibrium (LD) block known to be associated with HCV clearance in European populations. These SNPs were genotyped in 261 HCV-infected Egyptian subjects (130 with spontaneous clearance and 131 with chronic infection). The most associated SNPs were rs12979860 (P = 1.6×10−7) and the non-synonymous IL28B SNP, rs8103142 (P = 1.6×10−7). Interestingly, three SNPs at the two bounds of the region were monomorphic, reducing the size of the LD block in which the causal variants are potentially located to ∼20 kilobases. HCV clearance in Egypt was associated with a region of IL28B smaller than that identified in European populations, and involved the non-synonymous IL28B SNP, rs8103142.Spontaneous clearance of hepatitis C virus (HCV) occurs in ∼30% of acute infections. Host genetics play a major role in HCV clearance, with a strong effect of single nucleotide polymorphisms (SNPs) of the  IL28B gene, which encodes IFN-λ3, on spontaneous HCV clearance reported in populations of different backgrounds, mostly infected with viral genotypes 1 and 3  infects 170 million people worldwide and is thus a major Public Health problem IL28B, rs12979860, and the spontaneous clearance of HCV genotype 4 in an Egyptian sample IL28B SNPs in the spontaneous clearance of HCV in a larger Egyptian cohort. We found that the clearance effect was restricted to a region of IL28B smaller than that identified in European populations and that this effect also involved rs8103142, a non synonymous IL28B SNP.Egypt has the highest prevalence of HCV infection of any country in the world, with a seroprevalence ranging from 10% in children to 45% in adults http://www.unaids.org/en/regionscountries/countries/egypt) as well as among injection drug users (∼0.6%) This study combined data from two Egyptian cohorts. The first included 3994 individuals from a large epidemiological study in a village of the Nile delta, the details of which have been described elsewhere IL28B genomic region (IL28B) of these three SNPs, using the European CEU population available data from HapMap (http://www.hapmap.org) and 1000 Genomes project 2>0.3, with any of the three initial SNPs. Within the chromosome 19 region defined by these two SNPs, we included three additional regularly spaced SNPs  in LD, with an r2>0.3, with any of the three initial SNPs. Finally, we also selected the non synonymous SNP rs8103142 not present in the public databases but reported to be in strong LD with rs12979860 in European populations P-value<0.005 or low-quality genotype clustering. No individuals were excluded on the basis of genotyping call rate.We genotyped a total of nine SNPs within the c region . We firshttp://www.broad.mit.edu/mpg/haploview) software.Univariate tests of association between the genotyped SNPs and the clearance/persistence status of HCV infection were carried out using PLINK software IL28B region were monomorphic, whereas the frequencies of the other SNPs were similar to those observed in European populations. Univariate association analyses showed that the six polymorphic SNPs in the Egyptian population were strongly associated with spontaneous HCV clearance (−7] and rs8103142 . Association analyses conducted for these two SNPs separately in our two cohorts provided very similar results in terms of allele frequencies and odds-ratios indicating that possible HBV co-infection in some patients of our first cohort could not affect these results. In our whole sample, and taking rs8103142 as an example, the proportion of subjects with HCV clearance was much higher in subjects carrying the protective TT genotype (66% of clearers) than in those with CT (39% of clearers) or CC (23% of clearers) genotypes  was introduced into the model, the other five SNPs ceased to be significantly associated with clearance, confirming that this association was based on a single signal. We then estimated pairwise LD between the six polymorphic SNPs. The LD pattern of these SNPs, and that of rs12979860 in particular see , in thispulation . Interesn  haplotypes (one of which was much more common than the other) and five “at risk” haplotypes (conferring a predisposition to chronic infection) . The twoIL28B variants in the spontaneous clearance of HCV genotype 4 infection in an Egyptian population. In particular, the magnitude of the association with rs12979860 observed is similar to that reported in many studies conducted on HCV genotype 1 infection IL28B SNPs, showed that the LD block of the IL28B SNP cluster associated with HCV clearance was shorter than that in European populations. In particular, this analysis reduces the chromosomal region in which the causal SNPs are potentially located to about 20 kilobases of the IL28B genomic region, excluding the IL28A genomic region. Finally, both our univariate and haplotype analyses clearly showed that the core association depended on two SNPs, rs12979860 and rs8103142. SNP rs8103142 replaces a lysine residue by an arginine residue at position 70 (K70R) in IFN-λ3, a substitution predicted to be benign by Polyphen IL28B involvement in HCV infection.These results confirm the major role of Figure S1Linkage disequilibrium pattern for the six polymorphic SNPs in the Egyptian and European populations. Panel A shows linkage disequilibrium (LD) in terms of r2 values between rs12979860 and the eight other genotyped SNPs in two different populations (Egyptian/European). European r2 values were estimated from the CEU data from Hapmap and the 1000 Genomes project, Egyptian values were estimated from our overall sample. No data for rs8103142 were available for the CEU population, so the r2 value for this SNP could not be estimated for the European population. The SNPs rs955155, rs10424607 and rs576832 were monomorphic in the Egyptian population, and it was therefore not possible to estimate r2 values for these SNPs in the Egyptian population. Panel B shows the pairwise r2 for combinations of the six polymorphic SNPs of the IL28B genomic region in both the Egyptian population  and the CEU European population . No data for rs8103142 were available for the CEU population. Differences of less than 10% between the two r2 values are indicated in dark grey; differences of less than 25% are shown in light grey and differences of more than 25% are shown in white.(DOC)Click here for additional data file.


L'ostéoporose endocrinienne devrait devenir rare aujourd'hui puisque les endocrinopathies ont bénéficié d'un diagnostic plus précoce. Cependant elle reste fréquente, sous diagnostiquée et peu prise en charge. Sa gravité est essentiellement liée au risque fracturaire, et au risque élevé de morbi-mortalité. Le but de notre travail est de déterminer le profil ostéodensitométrique des patients suivis pour endocrinopathie et de définir les caractéristiques de l'ostéoporose et de l'ostéopénie chez ces patients. Il s'agit d'une étude de type transversale, descriptive portant sur 63 patients suivis pour endocrinopathies au service d'Endocrinologie-Diabétologie du CHU Mohamed VI de Marrakech, s’étalant sur une période de 02 ans allant du début du mois de Janvier 2012 au mois de Janvier 2014. La moyenne d’âge des patients était de 36,30 ± 15,38 ans, avec un sex-ratio de 0,31. Les pathologies endocriniennes étaient dominées par l'hypogonadisme (32%), et par l'hypercorticisme (19%), suivis par l'hyperthyroïdie (11%). Le T-score moyen au niveau du rachis et au niveau du fémur était respectivement de -1,65% ± 1,88 DS, et -0,85 ± 1,70 DS. L'atteinte de la densité minérale osseuse a été plus fréquente au niveau du rachis . L'ostéoporose a été surtout constatée au cours des hypogonadismes, hypercorticismes, hyperthyroïdies et hyperparathyroïdies. Néanmoins elle a été observée également au cours des hypothyroïdies et d'autres étiologies non connues pour être responsables d'une ostéoporose. Ce travail nous a ainsi conduits à situer l'intérêt de rechercher systématiquement l'atteinte osseuse devant toute endocrinopathie susceptible de provoquer une atteinte rhumatologique. La plupart des pathologies endocriniennes ont un retentissement osseux qui est pris en compte dans la morbidité de ces maladies. En effet, le tissu osseux fait l'objet d'un perpétuel remodelage sous contrôle, entre autres, de facteurs hormonaux. Ainsi, la plupart des désordres endocriniens s'accompagnent de remaniements de l'appareil ostéo-articulaire. Il s'agit principalement de l'ostéoporose . Cette dIl s'agit d'une étude transversale, à visée descriptive étendue sur une période de 02 ans allant du début du mois de Janvier 2012 au mois de Janvier 2014, intéressant les patients suivis pour endocrinopathie au service et en consultation d'Endocrinologie-Diabétologie du CHU Mohamed VI de Marrakech. Les critères d'exclusion sont: les patients suivis pour un rhumatisme inflammatoire et les patients suivis pour une maladie systémique. La saisie et l'analyse des données ont été réalisées à l'aide du logiciel SPSS version 10.Caractéristiques démographiques et cliniques: La moyenne d’âge de nos patients était de 36,30 ± 15,38 ans, avec des extrêmes de 12 à 73 ans. On note une nette prédominance du sexe féminin, avec un Sex-ratio de 0,3. La moyenne de l'indice de masse corporelle des patientes était de 25,12 ± 6,22 Kg/m2. Tandis que celle des patients était de 23,72 ± 7,28 Kg/m2. Les femmes ménopausées représentaient 27,1% de la population de l’étude. Les autres comorbidités sont représentées par l'hypertension artérielle , le diabète , la dyslipidémie  et le tabagisme .Nature de l'endocrinopathie: le motif de consultation des patients était dominé par l'aménorrhée primaire et secondaire (16%), le syndrome de cushing (16%) et les hyperthyroïdies (10%). Le délai entre le début des symptômes et la première consultation était de 5,5 ans ± 8,3 ans. Les pathologies endocriniennes chez cette population étaient très diversifiées. Elles étaient dominées par l'hypogonadisme rencontré dans 32% des cas et par l'hypercorticisme dans 19% des cas, suivis par l'hyperthyroïdie constatée chez 11% des patients , le T-score moyen au niveau du rachis était de -1,65% ± 1,88 DS le qualifiant d'ostéopénique, avec un T-score moyen fémoral de -0,85 ± 1,70 DS. L'atteinte de la DMO a été plus fréquente au niveau du rachis  avec une ostéoporose dans 36,5% des cas et une ostéopénie dans 25,4% des cas. L'atteinte fémorale a été surtout une ostéopénie, présente chez 26,4% des patients avec une ostéoporose chez seulement 17,5% des cas   .Au cours de l'hypogonadisme: les anomalies de la DMO rachidienne ont été constatées dans 75% des cas avec hypogonadisme, avec essentiellement une ostéoporose dans 45% des cas. Cette dernière avait concernée 13,3% des cas au niveau du fémur, avec une ostéopénie fémorale dans 40% des cas.Au cours de l'hypercorticisme: chez les patients avec hypercorticisme, l'atteinte de la DMO rachidienne avait concerné 66,7% des patients, avec une ostéoporose dans 50% des cas et une ostéopénie dans 16,7% des cas. L'atteinte de la DMO fémorale a été constatée chez 33,3% des patients.Au cours de l'hyperthyroïdie: le pourcentage de l'ostéoporose rachidienne a été de 33,3% au cours de l'hyperthyroïdie, tandis que l'ostéopénie rachidienne a été observée chez 33,3% des patients. Au niveau du fémur, l'ostéopénie a été constatés chez 66,7% des patients.Au cours de l'hyperparathyroïdie: parmi les patients ayant une hyperparathyroïdie, 33,3% avaient une ostéoporose rachidienne, et 33,3% avaient une ostéopénie rachidienne. L'atteinte de la DMO fémorale a été constatée chez 66,7% des patients.Au cours des autres endocrinopathies: l'analyse de l'atteinte de la DMO dans les autres endocrinopathies avait montré une ostéoporose dans 25% des cas d'hypothyroïdie, dans 50% des cas phéochromocytome, et dans 22,2% des cas d'hyperparathyroïdie. L'ostéopénie a était constatée respectivement chez 37,5%, 25% et 22,2% des patients.La chronologie d´apparition de l'atteinte de la DMO: l'atteinte de la DMO a été diagnostiquée essentiellement après ou simultanément à la découverte de l'endocrinopathie, par contre elle en était révélatrice dans 14% des cas  et des ostéoblastes (formation). Cette balance formation-résorption est placée sous l'influence de nombreux facteurs, notamment nutritifs et endocriniens . De nombCertaines ont un effet anabolique sur le tissu osseux en favorisant l'action des ostéoblastes ou en inhibant celle des ostéoclastes: l'hormone de croissance (GH) et l'insulin-like growth factor 1 (IGF1), l'insuline et les stéroïdes sexuels  . D'autreIl s'agit des prostaglandines dont la PGE2, des interleukines, de facteurs de croissance épidermiques qui sont des acteurs locaux activateurs des ostéoclastes. À l'inverse, le monoxyde d'azote (NO) inhibe la résorption osseuse .L'activité physique a également un effet sur le remodelage osseux. Les forces de déformation et de pression modifient l'activité de remodelage osseux. Une ostéopénie est d'ailleurs observée après un séjour en microgravité .L'ostéoporose cortisonique est la première cause des ostéoporoses secondaires. Longtemps négligée, son importance grandit, notamment dans le cadre de nouvelles indications de corticothérapie au long cours. En effet Il existe un lien épidémiologique certain entre la corticothérapie et les fractures. Certaines études estiment même que des fractures surviennent chez 30 à 50% des patients sous corticoïdes au long cours. Enfin, l'excès de corticoïdes favorise l'apparition d'un hypogonadisme et la diminution de production d'androgènes surrénaliens [L'hypogonadisme est la deuxième cause d'ostéoporose secondaire après la corticothérapie . Il reprLa PTH stimule la résorption osseuse et donc le flux de calcium et de phosphate de l'os vers le plasma. Elle augmente la réabsorption tubulaire du calcium et diminue celle des phosphates. Elle stimule enfin la 1’-hydroxylase rénale dans le tubule proximal. L'hypersécrétion de PTH par les glandes parathyroïdes est la conséquence d'une production excessive, inappropriée de PTH ayant pour principale conséquence métabolique une hypercalcémie. La plupart des hyperparathyroïdies sont asymptomatiques, de découverte fortuite au cours d'un bilan biologique ou d'une échographie cervicale . L'hyperLes hormones thyroïdiennes T3, T4) sont des activateurs du métabolisme de base. Leur sécrétion est placée sous le contrôle de la TSH hypophysaire, elle même placée sous l'influence de la sécrétion hypothalamique de TRH. Chez l'enfant, elles jouent principalement un rôle dans la croissance et la maturation du tissu osseux . L'ostéo, T4 sontLes autres endocrinopathies étaient très peu représentées dans notre série avec un effectif insuffisant pour pouvoir en déduire des conclusions.L'atteinte de la DMO au cours des pathologies endocriniennes est fréquente mais reste sous diagnostiquée et peu prise en charge. Sa gravité est liée au risque accru de complications, entre autres une augmentation du risque fracturaire, qui est un puissant facteur de risque de morbi-mortalité prouvé dans la littérature. La grande fréquence en population générale et l'importance du retentissement osseux des hyperparathyroïdies, hyperthyroïdies et des situations d'hypogonadisme et d'hypercorticisme les placent parmi les pathologies endocriniennes pour lesquelles l’évaluation osseuse est la mieux connue. Ce travail montre une fréquence élevée de l'ostéoporose et de l'ostéopénie chez les patients suivis pour des pathologies endocriniennes, même les moins connues pour l'avoir entrainée. Il nous a ainsi conduits à situer l'intérêt de rechercher systématiquement l'atteinte osseuse devant toute endocrinopathie susceptible de provoquer une atteinte rhumatologique. Mais aussi l'intérêt d'un bilan étiologique bien conduit, à la recherche d'une pathologie endocrinienne devant toute ostéoporose ou ostéopénie.


Le but de notre travail est de déterminer le profil des auto-anticorps chez 30 patients ayant un lupus systémique avec ou sans atteinte rénale afin d’établir une corrélation clinico-immunologique entre la néphropathie lupique et ces auto-anticorps. Il s'agit d'une étude transversale de 30 patients atteints de lupus érythémateux systémique diagnostiqués au service de dermatologie durant la période de Décembre 2010 à Décembre 2012 et réalisée conjointement avec le laboratoire d'immunologie. Les anticorps anti-ADN étaient retrouvés chez 17 patients (56.7%) suivis des anti-SSA dans 12 cas (40%). Cinq patients (62.5%) ayant une atteinte rénale avaient des anticorps anti DNA négatifs. Parmi ces patients avec atteinte rénale, 37.5% avaient des anticorps anti SSA sans anticorps anti DNA. La moitié des patients ayant une atteinte rénale (50%) avaient des anticorps anti SSA positifs. Notre série montre l'importance des anticorps anti-SSA surtout chez des patients avec des anticorps anti-DNA négatifs non seulement pour le diagnostic du lupus systémique mais aussi pour déceler certaines manifestations systémiques comme l'atteinte rénale. Le lupus érythémateux systémique (LES) est une maladie auto-immune caractérisée par la production d'un large panel d'auto anticorps: les anticorps antinucléaires particulièrement les anticorps anti-DNA natifs. De nombreuses publications ont établi une corrélation clinico-immunologique de patients atteints de LES, mais peu d’études se sont intéressées de manière précise à la prévalence des différents auto-anticorps en cas d'atteinte rénale. En effet, la néphropathie lupique est fréquente dans l’évolution du lupus systémique (20 à 50% des cas selon les séries) et influence considérablement le pronostic fonctionnel et vital de cette pathologie . La survIl s'agit d'une étude transversale de 30 patients atteints de lupus érythémateux systémique diagnostiqués au service de Dermatologie au CHU Ibn Rochd de Casablanca. Elle a été réalisée conjointement avec le laboratoire d'immunologie et a été menée sur une période de deux années (Décembre 2010-Décembre 2012). Les patients remplissaient au minimum quatre critères de l'American Collège of Rheumatology (ACR) [Parmi les 30 patients, 26 étaient de sexe féminin et 4 de sexe masculin avec un sexe ratio H/F de 0.15. La moyenne d’âge était de 35 ans avec des extrêmes de 14 à 59 ans. Les manifestations cutanées étaient observées chez 29 malades (96.7%) et l'atteinte hématologique chez 28 patients . Huit patients avaient une atteinte rénale soit 26.7%. La néphropathie lupique a été classée de type III de l'OMS dans 2 cas, de type IV dans 4 cas et de type V dans 2 cas. Les AAN ont été positifs chez 25 des 30 patients, avec un aspect moucheté (40%) et homogène (23.3%). Les anticorps anti-ADN étaient retrouvés chez 17 patients (56.7%) suivis des anti-SSA dans 12 cas (40%), anti SM/RNP dans 7 cas (23.3%), anti SSB et anti nucléosome chez respectivement 6 cas (20%), anticorps antihistone dans 3 cas (10%) et anticorps anti SM dans 2 cas (6.7%). Cinq patients (62.5%) ayant une atteinte rénale avaient des anticorps anti DNA négatifs . Parmi cDans cette série, la moitié des patients atteints de lupus systémique n'avaient pas d'anticorps anti-DNA. Dans ce groupe, parmi ceux ayant une atteinte rénale, 37.5% avaient des anticorps anti-SSA positifs. Ainsi, l'apport des anticorps anti-SSA parait fondamental en cas de lupus systémique avec atteinte rénale. Certes, notre série se heurte au biais du faible échantillonnage, néanmoins; il s'agit d'une étude prospective menée dans un service de dermatologie ayant utilisé un panel d'auto-anticorps représentatif. Ceci a permis par conséquent une comparaison plus large et une corrélation clinico-immunologique entre différents auto-anticorps en matière de néphropathie lupique. Dans notre série, l'atteinte rénale était notée dans 26.7% des cas qui étaient dans leur majorité des stades avancés IV et V (75%). La fréquence de cette atteinte rénale dans notre contexte est en accord avec les données de la littérature (20 à 50% des cas selon les séries) . Cette aUn suivi régulier des patients lupiques avec anticorps anti SSA positifs surtout ceux avec des anti-DNA négatifs s'impose afin de déceler une atteinte systémique notamment rénale d'autant plus que cette manifestation est sévère dans notre contexte.


Cette étude visait à évaluer la performance du système de gestion logistique (SGL) des intrants de lutte contre le paludisme (ILP) dans le Département du Littoral, au Bénin, en 2017.Il s’agissait d’une étude transversale évaluative qui s’était déroulée en juin 2017. Elle portait sur les structures de stockage et de cession des ILP ainsi que le personnel impliqué dans leur gestion. La performance du SGL était évaluée à partir de la conformité observée pour les composantes et sous-composantes de la « Structure », du « Processus » et des « Résultats » par rapport aux normes ou standards définis par le Ministère de la Santé.Un total de 36 structures a été enquêté avec leurs cibles secondaires. Il résulte que 52,78% des dépôts répartiteurs réunissaient les conditions optimales de stockage des intrants alors que seulement 33,33% des agents chargés de la gestion des ILP étaient formés en gestion logistique. La performance du SGL des ILP était insuffisante . La structure, ainsi que le processus avaient une conformité Insuffisante par rapport aux normes , engendrant des résultats jugés mauvais . La sous-composante la plus insuffisante était le système d’information de la gestion logistique (SIGL).Cette étude met en évidence la place du SIGL pour une meilleure performance de la gestion des ILP. Une attention particulière devra être accordée à ce volet. Le paludisme est la maladie parasitaire la plus répandue dans le monde . En 2016Type d’étude: Il s’agit d’une étude transversale évaluative qui s’est déroulée dans le département du Littoral du 6 au 29 juin 2017.Population d’étude: Les cibles primaires de notre étude étaient constituées par les dépôts de pharmacie  et points de cession des ILP) des FS publiques et privées du département du Littoral. Les Cibles secondaires étaient: les gestionnaires des dépôts répartiteurs de zones (GDRZ), les responsables des entrepôts des FS, les commis à la vente des médicaments (au point de cession), les bénéficiaires des ILP, les responsables de FS et le chef du service de pharmacie et pharmacovigilance du programme national de lutte contre le paludisme (PNLP).Echantillonnage: Trente-deux FS ont été sélectionnés par tirage aléatoire simple, sur les 52 du département. Les responsables de ces FS, ainsi que les responsables de dépôts et commis des postes de cession ont été enquêtés. De même, les dépôts répartiteurs des quatre zones sanitaires (ZS) du département, ainsi que leurs responsables ont été également enquêtés. Un bénéficiaire, identifié par commodité, a été enquêté dans chaque FS.La performance du SGL des intrants de lutte contre le paludisme a été appréciée sur la base de la combinaison des scores de conformité de la structure, du processus gestionnaire et des résultats, par rapport aux normes du Ministère de la Santé , 7. La sLes données ont été recueillies par des enquêteurs formés. Les outils utilisés sont: un questionnaire pour les gestionnaires de DRZ, responsables d’entrepôts de FS, les commis à la vente et clients externes; un guide d’entretien pour les responsables de FS et le chef du service de pharmacie et pharmacovigilance du PNLP; une fiche de dépouillement des outils de gestion; une grille d’observation des magasins et entrepôts. Les outils ont été pré-testés dans deux FS du département de l’Atlantique avant l’enquête, puis corrigés au besoin.® et analysées avec le logiciel STATA.11. Les proportions de conformité par rapport aux normes ont été calculées globalement pour les 36 structures évaluées, par sous-composante, puis par composante.Les données ont été saisies à l’aide du logiciel ExcelL’enquête a été autorisée par le Ministère de la santé et les responsables départementaux ainsi que ceux de chacune des FS. Le consentement oral des participants a été obtenu avant leur implication dans l’enquête. Leur anonymat a été conservé lors du traitement des données.L’ensemble des 36 structures prévues ont été enquêtées, dont quatre dépôts répartiteurs, 32 entrepôts de FS, dont 12 publics, 18 privés et deux confessionnels. Par ailleurs, 32 responsables de FS, 32 commis à la vente des ILP et autres intrants, 01 chef du service de pharmacie et pharmacovigilance du PNLP et 32 bénéficiaires des ILP dans les FS ont été enquêtés.Disponibilité des ressources humaines: chaque DRZ est géré par un comptable qui assure aussi la comptabilité du bureau de zone. Au niveau des entrepôts des FS, on notait une à trois personnes chargée(s) de la gestion des médicaments et ILP. Les profils des responsables se présentent comme suit: 14 comptables ; 9 aide soignants ; 6 Infirmiers . Les autres  sont 3 et représentent 9,37%. Le nombre d’années médian d’expérience des agents dans la gestion des dépôts est de 5 ans . Dix agents (dont 8 du secteur public) sur les 36, soit 27,78% étaient à leur poste il y a moins de 3 ans.Formation du personnel en gestion logistique: des 36 agents de dépôts pharmaceutiques, 12 (dont 2 Gestionnaires de DRZ), soit 33,33% ont bénéficié d’une formation en gestion logistique des ILP. Parmi eux, 4 ont bénéficié d’une remise à niveau au cours des 3 dernières années. Il n’y a pas eu de nouvelle formation au cours de 3 dernières années.Supervision: des 36 agents enquêtés, 31, soit 86,11% ont déclaré avoir été supervisés au moins une fois au cours des trois derniers mois par leur hiérarchie. Les équipes de supervision venaient de la direction départementale de la santé (DDS), du PNLP ou parfois étaient des équipes conjointes.Infrastructures et moyens de transport des ILP: Dix-neuf structures (2 DRZ et 17 entrepôts de FS) sur les 36, soit 52,78% disposant de magasin réunissent au moins 80% des conditions optimales pour le stockage des ILP. Parmi ces dernières, on retrouve seulement cinq FS privées sur les 18 enquêtées.Outils de gestion de stocks des ILP: Vingt-trois (23) structures sur les 36, soit 63,89% disposaient de 100% des fiches de gestion de stock des 12 ILP à l’étude. Le remplissage des fiches était à jour dans 47,22% des cas, soit 17 structures sur les 36.®) était disponible au niveau des quatre DRZ. Trois GDRZ sur 4 étaient formés à son utilisation. Il était fonctionnel et utilisé pour les activités dans deux DRZ sur les 4.Trois DRZ sur les quatre et 8 entrepôts de FS sur les 32 enquêtés (25%) avaient transmis les rapports des 6 derniers mois en respectant le délai et le rythme (la promptitude). La complétude des rapports des FS au niveau des DRZ pour les 6 derniers mois était de 67,91% soit 67 rapports reçus sur 110 attendus. Dans 21 structures sur 36, soit 58,33% on notait une discordance entre les données du rapport et celles de l’inventaire physique au moment de l’élaboration des rapport/commandes. Le logiciel de gestion  des structures disposaient de la liste des ILP retenus par la politique nationale et s’y réfèrent pour les commandes. La quantification des besoins en ILP se fait à chaque niveau de la pyramide et 77,78 % des responsables de la gestion des ILP en sont responsables. Les niveaux minimum et maximum des différents ILP ont été définis dans 14 structures sur les 36 enquêtées, soit 38,89%. Selon les GDRZ et des entrepôts des FS la faible disponibilité des intrants au niveau central limite leur application.Nous avons trouvé que 66,67% des structures réunissaient 80% et plus de critères normatifs de stockage des ILP dans les magasins. Toutes les structures (100%) respectent le stockage des périmés mais 31 sur 36, soit 86,11% avaient encore des intrants périmés dans les FS.La disponibilité des ILP a été appréciée sur douze intrants traceurs. Dans les DR, la disponibilité des ILP dans les DRZ le jour de l’enquête et au cours des 6 derniers mois est résumé sur la Au cours des six derniers mois, le taux de disponibilité des ILP dans les DRZ variait de 0% (pour l’Artéméther 20 et 80 mg) à 100% (pour la SP). Dans les FS ce taux variait entre 11,70% pour Artéméther 20 mg et 86,36% pour la Quinine injectable 600 mg .Un gestionnaire de DRZ sur les 4 et 28,12% des responsables d’entrepôts de FS ont déclaré avoir été satisfaits de la livraison des quantités d’ILP commandées. Sur 32 clients externes interrogés, 22 ont déclaré être satisfaits de la disponibilité des ILP dans les FS ; ils représentent 68,75%. Vingt et neuf d’entre eux, soit 90,63% trouvaient que les prix des intrants vendus étaient supportables.La conformité des sous-composantes de la « Structure » variait de 47,30% pour le SIGL, à 76,85% pour « l’infrastructure et les moyens de transport ». La conformité de la sous-composante « Technologie » était à 75%; celle des « outils de gestion », des « ressources financières » et des « Ressources Humaines » étaient respectivement de 73,73%, 68,33%, et 53,09%. L’appréciation de la performance de la composante « Structure » est insuffisante avec conformité de 63,20% du total attendu . La confCette étude a permis d’apprécier le niveau de performance du SGL des ILP dans le département du Littoral en 2017. Elle a identifié comme point de faiblesse majeur, l’insuffisance de mise en œuvre du SIGL. Les 36 structures (32 entrepôts de FS et 4 DRZ) prévus ont été investigués, avec leurs cibles secondaires. Elles représentent 60% des FS du département. Les 32 entrepôts, sélectionnés de manière aléatoire, regroupaient aussi bien des FS publiques que privées. La méthode d’échantillonnage ainsi que l’effectif des cibles, garantissent donc la représentativité des résultats à l’échelle du département.La sous-composante ressources humaines a été appréciée « Insuffisante » avec 53,09% du score attendu, compte tenu des insuffisances notées en termes d’effectif (surtout au niveau des DRZ), de qualification, de formation et d’organisation. Au niveau des DRZ il n’y avait qu’un seul agent, le comptable, ayant par ailleurs d’autres charges au bureau de zone, alors qu’il devrait être secondé par un magasinier ou un logisticien. Seulement 33,33% des agents ont bénéficié d’une formation en gestion logistique des ILP. Cette proportion est en deçà des résultats obtenus par Kolapo et al.  en RépubIl résulte de l’étude que 52,78% des structures enquêtées disposaient de magasin réunissant au moins 80% des conditions optimales pour le stockage des ILP et autres intrants/médicaments. En RDC, 64,2% la proportion des FS remplissait au moins 80% des conditions de stockage de produits contraceptifs . Au NigéCette sous-composante a été appréciée « mauvaise » avec 47,30% du score attendu. Des points faibles du SIGL observés sont: la faible promptitude et complétude des rapports des FS et DRZ; une faible proportion de gestionnaires des ILP formés sur le remplissage des outils du SIGL  avec pour corollaire des insuffisances dans le remplissage des fiches de gestion de stock. Cette situation prévalait aussi bien dans les FS publiques que privées et était d’autant plus important que les structures privées sont les plus nombreuses et très faiblement impliquées dans les activités de lutte contre le paludisme. Diagne et al. en 2012 . Ce mauvLa quantification des besoins en ILP se faisait par les structures elles-mêmes dans 77,78% sous la responsabilité des gestionnaires des ILP. Mais le processus a des défaillances liées à la faible disponibilité des données du SIGL pour la prise de décisions. En effet dans notre étude, seulement 47,67% des structures utilisaient les données du SIGL pour l’estimation des besoins. Contrairement aux observations de Adino et al. en 2011 à Addis-Abeba , les quaDans la présente étude, 63,89% des structures disposaient de 100% des fiches de gestion de stock des 12 ILP évalués. La complétude des fiches était bonne dans 47,22% des structures enquêtées. Selon Ouédraogo et al. [Le jour de la visite, tous les DRZ et toutes les FS étaient en rupture d’au moins un intrant. La diversité des intrants, leur faible disponibilité au niveau des agences de la CAME, les insuffisances dans l’utilisation des outils de gestion par les FS, le manque de rigueur dans le suivi des consommations des intrants à tous les niveaux sont autant d’éléments susceptibles d’expliquer ces résultats. La faible disponibilité des intrants dans les agences de la CAME s’exprime par la non satisfaction des quantités et des différents types d’intrants commandés par les DRZ. Des auteurs ont diversement obtenu d’autres résultats. Ouédraogo et al en 2006 en RDC  ont trouIl se dégage des relations de cause à effet entre les résultats des différentes sous-composantes dans notre étude. En effet, l’insuffisance en nombre et en formation des ressources humaines chargées de la gestion des ILP, couplée à des insuffisances dans leur supervision peut avoir pour conséquences les faiblesses dans l’exécution des différentes fonctions du cycle logistique, notamment la mise en œuvre du SIGL. Ces faiblesses vont se répercuter dans la disponibilité des intrants. La non-conformité des magasins de stockage aura des effets négatifs sur la qualité des intrants à offrir aux bénéficiaires, l’ensemble engendrant une performance globale du SGL des ILP jugée insuffisante.La présente a révélé que la performance globale du SGL des ILP dans le département du Littoral en 2017 était « Insuffisante ». Des points faibles les plus importants identifiés portaient sur le SIGL, avec pour corollaires, la faible disponibilité des intrants ainsi que la faible disponibilité et utilisation des données du système pour les prises de décisions. Vu l’importance des intrants dans la lutte globale contre le paludisme, la mise en place dans les centres de santé y compris le secteur privé, d’une démarche qualité portant sur le SIGL ainsi que le développement d’un système de veille pour l’identification et la destruction des circuits parallèles d’approvisionnement et d’introduction d’intrants hors la liste officielle sont nécessaires.Il existe des dysfonctionnements dans la gestion des ILP au Bénin comme dans plusieurs autres pays en Afrique, entrainant des péremptions et des ruptures de stock paradoxales;Plusieurs acteurs sont impliqués dans la gestion des ILP dans le système, entre autres le PNLP, la centrale d’achat des médicaments essentiels, les partenaires techniques et financiers, les ZS, les FS aussi bien publiques que privées;Un circuit de l’information sur l’approvisionnement et l’utilisation des ILP est défini et diffusé au niveau des différents acteurs.Cette étude démontre que la performance du SGL des ILP dans le département du Littoral au Bénin était « Insuffisante »; cette situation constitue un obstacle à la lutte efficace contre le paludisme;Le développement d'un SGL des ILP efficace pour la lutte contre le paludisme nécessite: la mise en place dans les centres de santé y compris le secteur privé, d’une démarche qualité dans la collecte et l’utilisation des données sur le système logistique de lutte contre le paludisme;Le développement d'un système de veille pour l’identification et la destruction des circuits parallèles d’approvisionnement et d’introduction d’intrants hors la liste officielle dans les stocks d’intrants de lutte contre le paludisme.Les auteurs ne déclarent aucun conflit d'intérêts.


Staphylococcus aureus bacteria lacking the SPase I SpsBthat are viable and able to grow in vitro when over-expressinga native gene cassette encoding for a putative ABC transporter. This transporterapparently compensates for SpsB's essential function by mediating alternativecleavage of a subset of proteins at a site distinct from the SpsB-cleavage site,leading to SpsB-independent secretion. This alternative secretion system alsodrives the main mechanism of resistance to an arylomycin-derived SpsB inhibitor,by means of mutations in a putative transcriptional repressor(cro/cI) causing over-expression of the ABC transporter.These findings raise multiple interesting biological questions. Unraveling themechanism of SpsB-independent secretion may provide an interesting twist to theparadigm of bacterial secretion.Secretion of proteins into the extracellular environment is crucial for thenormal physiology and virulence of pathogenic bacteria. Type I signal peptidase(SPase I) mediates the final step of bacterial secretion, by cleaving proteinsat their signal peptide once they are translocated by the Sec or twin-arginine(Tat) translocon. SPase I has long been thought to be essential for viability inmultiple bacterial pathogens. Challenging this view, we and others have recentlycreated  Staphylococcus aureus is an important human pathogen that can causelife-threatening invasive infections, such as bacteremia, endocarditis, pneumonia, andosteomyelitis. Because of its nominal essentiality, Type I signal peptidase (SPase I)has been investigated as a potential antibacterial target. Several factors makeS. aureus SpsB an attractive antibiotic target S. aureus have been generated 2cro/cI confer resistance to arylomycin-derived SpsB inhibitors inS. aureus3S. aureuscro/cI locus are associated with over-expression of the 3-genelocus immediately downstream of cro/cI which encodes a putative ABCtransporter 3S. aureus lethality caused by eitherpharmacologic inhibition or genetic ablation of SpsB, and is associated with secretionof alternatively cleaved proteins 3S. aureus SpsB in vitro. Togetherthese findings introduce a new concept, which challenges the common belief that SpsB isabsolutely essential for viability, and which invites a number of interestingquestions.A surprising observation from resistance studies performed by our group and others isthat mutations in the locus encoding for the putative transcriptional regulatorS.aureus strain and an S. aureus strain that lacks SpsBwhile over-expressing the ABC transporter show that only a minor subset of proteins thatare normally SpsB-cleavable is alternatively secreted 3First, to what extent does the putative ABC transporter compensate for the absence offunctional SpsB? Proteomic studies of the secretome of a wild-type invitro, it is unable to establish infection in a mouse model S. aureus virulence factors in vitro bacterial growth from in vivoinfectivity. An interesting consideration that follows is that since S.aureus is a commensal of mammalian skin and nares The observation that the alternative cleavage occurs N-terminal of the SpsB cleavage sitesuggests the involvement of a membrane-localized protease. Although the strain lackingSpsB and over-expressing the ABC transporter appears to grow well in vitro? To speculateon the answer, it would be relevant to understand the mechanism by which SpsB inhibitionleads to bacterial death. Two main hypotheses, that are not mutually exclusive, can beproposed. First, in the absence of SpsB activity, cell death may be caused byaccumulation of unprocessed proteins leading to disruption of membrane integrity. Thishypothesis raises the possibility that the ABC transporter is directly or indirectlyinvolved in removal of unprocessed proteins. It is possible that the transporter enablesa hypothetical membrane-associated proteolytic enzyme to degrade these unprocessedmembrane proteins. Second, under SpsB inhibitory conditions, cell death may occurbecause certain secreted proteins that are essential for viability cannot be secreted.In the context of this hyopothesis, it can be speculated that in the absence of SpsB,the ABC transporter enables secretion of an individual protein or a combination ofproteins that is otherwise essential. Although such proteins have not been identified asyet, this possibility also cannot be excluded. Future experiments to elucidate themechanism by which the ABC transporter compensates for the essentiality of SpsB shouldhelp address this question.Second, by which mechanism does the alternative ABC transporter-mediated secretionpathway compensate for the essentiality of SpsB cro/cI mutations) rendersS. aureus much more sensitive to treatment with anarylomycin-derived SpsB inhibitor in mice cro/cI itself is a likelycandidate. It is possible that cro/cI is part of an as yet to bedefined signaling mechanism that senses the presence of arylomycin in a naturalenvironment.Third, what is the physiological role of the putative ABC transporter? Over-expression ofthe ABC transporter to overcome a loss of secretion may be a protective mechanism in anatural environment in which arylomycin-like antibacterials are present, and this couldbe relevant for niche competition. In this context, it is noteworthy that inactivationof the ABC transporter  are found as an intact collinearunit in 133 of 145 Staphylococcus genomes examined. However, the operonis disrupted in a subset of strains in at least two pathogenic staphylococcal species.First, in two S. epidermidis genomes, the genes encoding ABC2 and ABC3are missing entirely . The loss of these two genes likely reflects the lackof a functional ABC transporter, as each is essential for resistance of a S.aureus USA300 cro/cI mutant to SpsBinhibitors S.aureus genomes analyzed, the operon has been interrupted by the obviousinsertion of an ~15 kb transposable element between the ABC2 and ABC3 encoding genes. While the gene encoding ABC3 in these strains may no longer be under thecontrol of the cro/cI promoter, it is unclear whether it can still beexpressed and whether an active ABC transporter can be formed. Thus, the operon is notuniversally intact in all Staphylococcus genomes; further experimentalstudies are needed to determine whether insertion of the transposable element in thecro/cI - ABC transporter operon inactivatesSpsB-independent secretion.Fourth, is the Cro/CI - ABC transporter operon universally conserved in pathogenicS.aureus that is able to bypass the nominal essentiality of SpsB raises anumber of interesting questions. Determining the molecular mechanism of this alternativesecretion pathway has the potential to provide new insights into the basic biology ofbacterial secretion and to aid design of new antibacterial therapies.In conclusion, the discovery of a potential new secretion system in


Cryptococcus gattii species complex harbors the main etiological agents of cryptococcosis in immunocompetent patients. C. gattii molecular type VGII predominates in the north and northeastern regions of Brazil, leading to high morbidity and mortality rates. C. gattii VGII isolates have a strong clinical relevance and phenotypic variations. These phenotypic variations among C. gattii species complex isolates suggest that some strains are more virulent than others, but little information is available related to the pathogenic properties of those strains. In this study, we analyzed some virulence determinants of C. gattii VGII strains  isolated from patients in the state of Piauí, Brazil. The C. gattii R265 VGIIa strain, which was isolated from the Vancouver outbreak, differed from C. gattii CG01, CG02 and CG03 isolates  when analyzed the capsular dimensions, melanin production, urease activity, as well as the glucuronoxylomannan (GXM) secretion. Those differences directly reflected in their virulence potential. In addition, CG02 displayed higher virulence compared to R265 (VGIIa) strain in a cryptococcal murine model of infection. Lastly, we examined the genotypic diversity of these strains through Multilocus Sequence Type (MLST) and one new subtype was described for the CG02 isolate. This study confirms the presence and the phenotypic and genotypic diversity of highly virulent strains in the Northeast region of Brazil.The  Cryptococcus neoformans–Cryptococcus gattii complex are the etiological agents of cryptococcosis, a life-threatening disease that affects the lungs and brains of humans and animals  of the species revealed five distinct molecular types: VGI, VGII, VGIII, VGIV, and VGIV/VGIIIc  obtained from Piauí, Brazil.in vitro assays were performed to measure the best characterized cryptococcal virulence factors. The production of melanin by C. gattii VGII isolates was assessed employing both spot plate and spectrophotometric assays. After 72 h of incubation of cryptococcal isolates serially diluted onto minimal media containing L-DOPA, we noticed that CG01, CG02, and CG03 isolates displayed increased rates of pigmentation when compared to R265 . In addition, we evaluated the laccase activity, by the quantification of melanin-like pigment upon exposure to L-DOPA; after 72 h of incubation, the supernatant was collected and the amount of melanin-like pigment was detected spectrophotometrically (OD405). In comparison to the R265 strain, CG02 cultures showed a fourfold increase in laccase activity, the CG01 cultures showed a twofold increase in laccase activity, and the CG03 cultures showed no significant differences in laccase activity . Urease activity was also analyzed for CG01, CG02, and CG03 isolates. Following the incubation of each strain in the Roberts urea broth, the amount of urease was quantified spectrophotometrically in the supernatant (OD560). The urease activity of CG01 and CG03 cultures were similar to that of R265, whereas the CG02 isolate displayed a slight increase in urease activity compared to R265 .To evaluate possible differences regarding the virulence between the isolates, Figure 2A). Determination of the capsule diameter ratio showed that all three strains had a larger capsule size compared to the R265 strain . The amount of secreted GXM was evaluated by ELISA. All three strains presented higher amounts of polysaccharide content in the culture supernatant compared to R265 strain . Capsular morphological analysis by scanning electron microscopy revealed a clear difference among capsular fibers of the isolate cells . Although the secretion of capsular components, such as GXM, is required for capsule assembly, capsule enlargement also requires polysaccharide molecules with higher effective diameters .Capsule enlargement and GXM secretion are essential factors for cryptococcal pathogenesis ; these fiameters . We thenC. neoformans-C. gattii species complex are facultative intracellular pathogens that can survive in macrophages .Members of the rophages . Since crophages , we assen = 5 animals/isolate) and compared host survival and fungal burden. Differences in virulence profile were observed for all investigated isolates. Mortality curves revealed that the CG02 isolate displayed a hypervirulent profile in the intratracheal model of infection (LT50 = 21 days)  and the intranasal inhalation model . In fact, the CG02 strain produced a higher pulmonary fungal burden overall after 10 and 15 days post-infection  and was the only isolate detected in the brain after 10 days of infection . Based on prior studies, the VGIIa R265 strain was chosen as a positive control for virulence  . The CG03 had an attenuated virulence profile then compared to R265 . Both CG01 and CG03 were unable to reach the brain . Lungs infected with R265, CG01, CG02 and CG03 isolates  all comprised mostly of cryptococcal cells (cryptococcomas).Phenotypic variations linked to virulence factors were observed for the investigated isolates and these were compared to R265 strain. To evaluate whether these phenotypes are directly related to the isolates virulence, mice were infected mice ( = 31.6) . CG01 haC. gattii VGII isolates in our study, we performed the ISHAM MLST consensus typing scheme. DNA samples were sequenced for all seven loci according to the ISHAM typing scheme (Table 1). The MLST profiling of the isolates revealed a clear genetic diversity. CG01 was classified as the sequence type 125 (ST 125), while CG03 belongs to the ST 127. A new subtype was described for CG02 isolate, which was named as ST 454.With the intent of evaluating the genetic diversity of the three g scheme . The seqg scheme  were useC. gattii species complex has attracted attention as a public health issue  demonstrated that virulence is related to the distinct characteristics of individual strains and is not specifically associated with a particular C. gattii species complex molecular type  from Piauí, Brazil. The pathogenic factors linked to these differences were mostly related to the surface architecture of C. gattii, including melanin synthesis, capsular structure and urease activity. The pathogenicity analyses were based on assays performed in vitro and in vivo with macrophages and mice, respectively. Additionally, it was demonstrated that the CG02 strain is hypervirulent in comparison to the R265 (VGIIa) strain; CG02 also has the ability to penetrate into the central nervous system (CNS), leading to the development of meningoencephalitis, which is not a common characteristic of C. gattii VGII species. The ability of CG02 strain to transpose the blood brain barrier must be one of the reasons as to why it displays a higher virulence profile when compared to R265 and other CG strains. Despite no information about the MAT locus identity of the isolates here described, a higher proportion of MAT alpha strains are present in Brazil , ST127 (CG03), and ST452 (CG02) indicated significant differences in the colonization of the lungs and brains of mice. ST125 (CG01) and ST127 (CG03) displayed a dominant pulmonary infection in mice and some phenotypic similarity with the isolate R265 (ST20-VGIIa). In comparison to members of the C. neoformans species complex, isolates of the C. gattii species complex usually cause lung infections, and their VGII strains are less efficient in spreading to the human brain  guideline. Mice were housed in groups of five in filtered top ventilated cages and were provided with food and water ad libitum. All efforts to minimize animal suffering were made. Before mortality analysis, mice were intraperitoneally anesthetized with 100 mg/kg ketamine and 16 mg/kg xylazine. The mice were analyzed twice daily for any signs of suffering, defined by weight loss, weakness, or the inability to eat or drink; they were sacrificed following any signs of suffering. The Universidade Federal do Rio Grande do Sul (UFRGS) Ethics Committee for Use of Animals (CEUA – 19801) approved the use of animals in the present work.C. gattii strains were used in the study. The first strain, R265 (ATCC MYA 4093), was previously identified  of molecular type/genotype VGII involved in this study were isolated from immunocompetent patients who had cryptococcal meningitis  . For subsequent experiments, minimal medium was used .Four different entified . The thr Brazil) . Such st560 nm) in urea broth were performed after a 4 h interval. All phenotypic assays were performed thrice. For capsule measurements, R265, CG01, CG02, and CG03 were grown in YPD for 24 h at 30°C and 200 rpm. The culture was then centrifuged for 10 min at 3,522 × g and the cells were washed twice with phosphate-buffered saline (PBS). Aliquots of 104 cells were incubated in RPMI-1640 medium  in a final volume of 500 μL in 24-well plate for 72 h at 37°C and 5% CO2. Cells were mixed with India ink and analyzed under microscope as described above. Cell and capsule sizes were measured using ImageJ software (NIH2). Total cell size was defined as the total diameter of the cell, including the capsule. Capsule size was calculated as the difference between the diameter of the total cell and the cell body diameter, defined by the cell wall. At least 100 cells were measured for each growth condition.Pigmentation, urease activity, capsule formation and ability to grow at 37°C were evaluated for each isolate. Melanin production was visually determined following the growth of cryptococcal cells, which had been serially diluted in solid minimal medium supplemented with 1 mM L-3,4-dihydroxphenylalanine (L-DOPA) and incubated for 72 h either at 30°C or 37°C . LaccaseStaining of the cryptococcal surface components (chitin and GXM) was performed as described . Fungal g . The supernatant fluids were collected and again centrifuged at 15,000 × g  to remove smaller debris. The pellets were discarded and the resulting supernatant was concentrated approximately 20-fold using an Amicon  ultrafiltration cell  supplemented with 10% Fetal Bovine Serum (FBS – Sigma) in each well of the 96-well culture plates (TPP). After 24 h of incubation (37°C and 5% CO2), the medium was replaced with fresh medium containing 1 × 106 cells of each fungal strain, obtained after a 18 h of growth in YPD and extensive washing in PBS and opsonization with anti-GXM antibody 18B7 . The plates were then further incubated (2 and 24 h at 37°C and 5% CO2). Yeast cells that were not internalized by the macrophages were removed with PBS washes. Fungal survival was evaluated after macrophage lysis with sterile ice-cold water and subsequent plating on YPD for Colony Forming Units (CFU) determination. This assay was performed in thrice for each strain.Assays were conducted to evaluate the susceptibility of the 5 yeast cells using an intranasal inhalation infection model. We also performed intratracheal infection of mice with 2 × 106 yeast cells/mL . The infection was performed and monitored twice daily for moribund signals. The median survival values were calculated by the Kaplan–Meier survival analysis. Animal studies were approved by the Federal University of Rio Grande do Sul Ethics Committee.Virulence studies were conducted as previously described . Fungal 5 yeast cells and monitored twice daily for moribund signals. Mice were euthanized by CO2 inhalation. The lungs and brain were homogenized in PBS, diluted and plated on YPD for CFU determination at each time point for fungal load determination .Fungal cells were cultured in 50 mL YPD medium at 30°C overnight with shaking, washed twice and resuspended in PBS. Groups of 5 female BALB/c mice (∼5 weeks old) were intraperitoneal anesthetized with Ketamine (100 mg/kg) and Xylazine (16 mg/kg) and then infected with 1 × 10URA5, CAP59, LAC1, GPD1, PLB1, SOD1) and the IGS1 region. For each locus studied, different genetic sequences present within a species are assigned as distinct alleles. The combination of the identified alleles at each of the loci defines the allelic profile or sequence type for each isolate. The data generated can be used to determine whether the fungal isolates are clonal or have undergone recombination. The URA5, IGS1, CAP59, LAC1, GPD1, PLB1, and SOD1 gene fragments were amplified using the published PCR conditions for all seven loci  were determined using the online C. gattii MLST database4. The new allele was submitted to the ISHAM MLST database for inclusion.Isolates were subtyped using MLST using partial sequence analysis of six housekeeping genes (ven loci . PCR proVB, LM, AF, JR, ES, GdSA, SF, CS, AS, LK, and MV prepared the experimental design. VB, LM, AF, JR, and ES conducted the phenotyping, CFU analysis, and animal experimentation. VB, GdSA, and SF performed the MEV, immunofluorescence and LS measurements. SF, CS, AS, LK, and MV provided reagents and equipment. VB, LM, AF, JR, ES, GdSA, SF, CS, AS, LK, and MV discussed the results and wrote and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Eugenol and its isomer isoeugenol are both used as flavouring agents or food additives in food products, and have both some similar biological properties. However, the difference in biological activities between eugenol and isoeugenol is rarely studied. In this study, the profiles of antioxidant, DNA-protective effects and antibacterial activities of eugenol and isoeugenol against several common foodborne pathogens were investigated and compared under various experiment conditions. Results showed that eugenol and isoeugenol had strong antioxidant activity, the protective effect against DNA damage and antibacterial activity. In addition, it was found that isoeugenol exhibited the higher biological activities mentioned above than eugenol, which was because isoeugenol had a carbon–carbon double bond closer to the benzene ring compared with eugenol. However, the specific reason needs to be further studied. Environmental factors can easily lead to food oxidation while microorganisms can easily result in food poisoning and food spoilage, which is one of the most important issues facing the food industry and consumers . AccompaEugenol , a natural phenolic compound found in essential oils from plants including clove, cinnamon, basil, and nutmeg, has been considered non-mutagenic, non-carcinogenic and generally recognized as safe (GRAS) by Food and Drug Administration , antibacTherefore, the aim of the present study was to investigate and compare the profiles of antioxidant, DNA-protective effects and antibacterial activities of eugenol and isoeugenol against selected foodborne pathogens through a variety of antioxidant and antibacterial methods under various experiment conditions, which is very important for the preparation of eugenol isomers and their application in food and medicine.2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,6-ditert-butyl-4-hydroxytoluene (BHT), 2,2ʹ-azobis (2-methylpropionamidine) dihydrochloride (AAPH) and 2,2ʹ-azino-bis (3-ethylbenothiazoline-6-sulfonic acid) diammonium salts (ABTS) were from Sigma (USA). 2, 4, 6-Tri (2-pyridyl)-s-triazine (TPTZ), eugenol and isoeugenol were purchased from Fluka (Switzerland). Propidium iodide (PI) was from BD Biosciences. The pBR322 plasmid DNA was from Takara Bio Co. Ltd. . Nutrient agar (NA) and nutrient broth (NB) mediums were from Beijing Aoboxing Bio-tech Co. Ltd. . Other chemicals used were all of analytical grade.Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 6051 and Listeria monocytogenes ATCC 19115. Three Gram-negative bacteria were Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 19430, and Shigella dysenteriae CMCC (B) 51252. Strains were provided by the College of Life Science, Shanxi Normal University, and cultured at 37°C on nutrient agar (NA) and nutrient broth (NB) mediums.Six kinds of common food spoilage bacteria are selected in the study. Three Gram-positive strains were 50 value, that is the effective concentration at which free radicals are scavenged by 50% and is obtained by interpolation from regression analysis.The scavenging rate and scavenging activity of the sample on DPPH radicals were determined as described in a previous report . The sca50 value were calculated using the equation described for DPPH assay.The ABTS cation radical scavenging activity was determined according to the method described by Xu et al. . The sca4 solution (100–1000 μM), and FRAP value was expressed as millimoles Fe(II) per gram sample.The reducing ability was determined by using FRAP assay described by Xu et al. . The staS × N/W. In this formula, S was the volume of sodium thiosulphate solution (blank corrected) in mL, N was the normality of sodium thiosulphate solution (0.02 N) and W was the weight of oil sample (gram).Lipid peroxidant value (POV) of samples was evaluated according to the method of Li et al. with some modiﬁcations . The sam2+ and H2O2 was estimated with the DNA nicking assay as described in the previous report [The ability of samples to protect supercoiled pBR322 plasmid DNA against Fes report .The ability of samples to protect supercoiled pBR322 plasmid against AAPH was measured according to the method described by Zhang and Omaye  with som7 CFU/mL of bacteria spread on nutrient agar (NA) medium. Oxford cups (6 mm in diameter) were placed on the inoculated agar, and then 100 μL of sample was added with a micropipette. The diameter of zone of inhibition (ZOI) was measured after 24 h of incubation at 37°C. Tests were performed in triplicate.The sample was first dissolved and then sterilized by filtration through 0.22 μm Millipore filters. Antibacterial tests were then carried out by the Oxford cup method using 100 μL of suspension containing 107 CFU/mL. The tubes were incubated at 37°C for 24 h and then examined for evidence of the growth. The MIC and MBC were determined according to the method described by Diao et al. [Two fold serial dilutions of samples were prepared in sterile NB medium. To each tube 100 μL of the exponentially growing bacterial cells was added to give a cell concentration of approximately 1 × 10o et al. .7 CFU/mL). The cultures were incu bated at 37°C and shaken at 120 rpm. At selected time intervals, samples from test culture were taken and the absorbance at 600 nm (OD600) was measured.One hundred microliters of samples filtrating through 0.22 μm Millipore filters was added to 4.8 mL of the sterile NB medium, and then mixed with 100 μL of a 10 h culture of tested bacteria  equipped with a CellQuest software (BD Biosciences) were used to analyse 1 × 104 cells after 30 min of reaction in a dark environment at room temperature. Cells were sorted into living and necrotic cells, and this assay was repeated five times.Logarithmic phase bacteria were collected by centrifugation at 6000×g for 5 min, washed three times, and resuspended in PBS (pH 7.4). Tested bacteria were treated with different concentrations of samples. After 0.5 and 1 h, cells containing approximately 1 × 10p < 0.05) between the means by Data Processing System  and Excel program.One-way analysis of variance (ANOVA) and Duncan’s multiple range tests were carried out to determine significant differences , indicating that the scavenging activity of isoeugenol was slightly higher than that of eugenol. However, the scavenging activity of eugenol and isoeugenol on DPPH radicals was lower than that of Trolox (EC50 was 13.5 μg/mL). The profile of scavenging activity of eugenol isomers on ABTS was similar to the result of the scavenging DPPH radicals. Somewhat differently, the EC50 values on scavenging ABTS cation radicals were 146.5 and 87.9 μg/mL for eugenol and isoeugenol, and the scavenging activity of isoeugenol was significantly higher than that of eugenol (p < 0.05), which was lower than of Trolox (EC50 was 84.34 μg/mL). These differences in data between DPPH and ABTS assays were likely due to different experimental conditions. The disappearance of DPPH and ABTS cation radicals is directly proportional to the amount of antioxidant present in the reaction mixture. Similarly, eugenol and isoeugenol showed a concentration-dependent scavenging of the DPPH and ABTS cation radicals at certain concentrations. Their antioxidant activity in the above assays may be mediated through direct trapping of the free radicals through transfers of hydrogen or electron [The scavenging activity of eugenol and isoeugenol on DPPH and ABTS radicals is shown in electron .Table 1.p < 0.05). These results suggested that eugenol isomers could result in reducing Fe3+/ferricyanide complex to the ferrous form (Fe2+), and had a remarkable potency to donate electron to reactive free radicals, transforming them into more stable non-reactive species and terminating the free radical chain reaction.The FRAP may serve as a significant indicator of the potential of antioxidant activity . Table 1p > 0.05) with the prolong of treatment time and only a 2.8% increase from days 10 to 30 , and increased from 1.2% to 23.4% and 67.2% for 60 min (p < 0.05) respectively. And under the same conditions, the necrosis rate of E. coli treated with isoeugenol dramatically increased to 14.7% and 80.8% for 30 min (p < 0.05) and increased to 28.6% and 92.7% for 60 min (p < 0.05) respectively. As expected, similar results were observed for L. monocytogenes cells treated with eugenol and isoeugenol after incubation with different times  was potently inhibited by eugenol and isoeugenol to the same extent [Eugenol and isoeugenol are the members of the phenylpropanoids class of chemical compounds, and there have been some studies on their antioxidant activity respectively ,5,9,10. e extent . The pre2+ and H2O2 has been reported by Nam et al. [Reactive oxygen species (ROS) are a major source of oxidative stress in cells and are generated by external and internal factors, resulting in the damage of proteins, carbohydrates, lipids and DNA. More and more evidence suggested that oxidative breakage of mammalian cellular DNA led to cell death, tissue damage and a wide array of neurological and pathological disorders . Previoum et al. , which wFrom the above results, eugenol and isoeugenol exhibited better antioxidant activities and DNA damage protective effect, which may be mediated through direct trapping of the free radicals or inhibiting the propagation of radical chain reactions through transfers of hydrogen or electron . The traE. coli and L. monocytogenes based on growth curve analysis and flow cytometry assays. Expectedly, similar to results of the antioxidant activity, isoeugenol also exhibited stronger antibacterial activity than eugenol as a whole. The phenolic hydroxyl group had weak acidity, which partly resulted in its inhibitory activity on bacteria. In addition, polyphenols perform multiple biological functions and many of these functions have been attributed to its antioxidant activity [Some studies have reported the antibacterial activity of eugenol against some bacteria ,28–30, bactivity , and a pIn conclusion, it is clear that eugenol and isoeugenol had an excellent reducing power and exerted antioxidant activity against DPPH, ABTS, lipid peroxidation, and possessed the protective effect against DNA damage induced by hydroxyl radical and AAPH, as well as antibacterial activities against several common foodborne pathogens. Their antioxidant activity and DNA damage protective effect were dose-dependent, increasing with a higher dosage in a certain concentration range, and the antibacterial effects were significantly influenced by incubation time and concentration. In addition, it was found that isoeugenol exhibited a higher antioxidant activity, DNA damage protective effect and antibacterial activity than eugenol, which was probably because isoeugenol had a carbon–carbon double bond closer to the benzene ring compared with eugenol, indicating that the biological activity of eugenol and isoeugenol mainly came from their structure, while subtle differences in the structure of eugenol isomers can lead to obvious change in their biological activity.