To investigate the relationship between plasma and cyst concentrations of albendazolesulphoxide (ASO) and their effects on parasitological findings and disease recurrence in patients with liver hydatidosis.The study was conducted at the University Hospital for Infectious Diseases “Dr. Fran Mihaljević,” Zagreb, Croatia, between August 2006 and January 2011. Consecutive patients  were treated with albendazole (3 × 5 mg/kg/d) over 28 days before surgical cyst removal (n = 34) or percutaneous evacuation (PAIR) (n = 14). Plasma ASO was determined on days 10 and 28 of treatment and cyst concentrations at surgery/PAIR.P < 0.001) and higher than cyst concentrations at the time of treatment . Higher cyst (but not plasma) concentrations were independently associated with lower odds of protoscolex motility  and higher odds of protoscolex destruction . With adjustment for age and protoscolex motility, higher day 10 plasma concentrations (but not cyst concentrations) were associated with lower odds of disease recurrence . Plasma concentrations did not predict cyst concentrations.Disease recurred in 3 surgically treated patients. Variability of ASO concentrations was substantial. Plasma concentrations on day 10 were higher than on day 28 (geometric means ratio [GMR] 2.00; 95%CI 1.38-2.91, Viability of protoscolices progressively decreased with increasing ASO concentrations in the cyst. Data strongly suggested that higher plasma concentrations reduced the risk of disease recurrence. E. granulosus, E. multilocularis, E. vogeli, and E. oligarthus  (still most commonly used method); percutaneous aspiration of the cyst with instillation of a scolicidal agent , ie, the PAIR  procedure, which seems to have greater clinical efficacy and lower rate of complications than the surgical procedure; and treatment with benzimidazole anthelmintic drugs. The latter, pharmacological option might be used as a monotreatment in the case of smaller cysts or when invasive approaches are not feasible, but it is typically adjunctive treatment to surgery or PAIR used to prevent dissemination of scolices from ruptured cysts. In this setting, benzoimidazoles are used over at least 4 weeks before the definitive treatment . Among bThis prospective observational prognostic study was conducted at a tertiary care university-affiliated teaching hospital between August 2006 and January 2011 and was approved by the institutional Ethics Committee. The only procedures that differed from the standard treatment/follow-up protocol were blood and cyst fluid samplings for determination of drug concentrations and laboratory evaluation of protoscolex viability.3/mm3).Eligible for inclusion were consecutive patients  with standard ,8 clinicPatients were hospitalized, routine biochemical/hematological tests and serological tests were performed, and 4-week treatment with albendazole was commenced . AlbendaTM, NovaTec GmbH, Leinfelden-Echterdingen, Germany]), and indirect fluorescence microscopy. For the latter analysis, after removal of fat using acetone, cryostat-madeprotoscolex slices were incubated (37°C for 30 minutes) with diluted sera (1:100 in phosphate buffer pH 7.2), rinsed (3 × 10 minutes), incubated  with anti-human IgG antibodies labeled with fluorescin-izocyanate , rinsed again, and analyzed (Sera were assessed for the presence of antibodies (IgG) against echinococcal antigens using a commercial semiquantitative enzyme-linked imunosorbent assay (ELISA) in line with the manufacturer’s instruction  method with UV detection (wavelength 293 nm) developed by Hoaskey et al  and modiAll evaluations were done by light microscopy at 37°C using unstained wet mount . Native 1) - ln(c2)〉] or regression coefficients (β). Within-subject variability of plasma concentrations  was determined as relative standard deviation (%RSD = 100 x √es2wr–1) 2. Parasitological: do plasma and/or cyst ASO concentrations affect parasitological outcomes? Proportions of patients with motile protoscolices/patients with morphologically changed protoscolices were analyzed by fitting generalized linear models and results were expressed as odds ratios (OR). 3. Clinical outcome: are ASO concentrations related to disease recurrence? This relationship was investigated by generalized linear models (effects expressed as OR). We used SAS 9.3 statistical software  licensed to Zagreb University School of Medicine.We posed three primary research questions. 1. Pharmacokinetic: what is the relationship between plasma and cyst concentrations? Analyses were performed by fitting general linear (mixed) models to ln(c) data and the effects were expressed as geometric means ratios, GMR [GMR = exp〈ln submitted to surgery and 14 to PAIR as the final treatment. Diagnosis was confirmed parasitologically in all enrolled patients. Their characteristics are summarized in P < 0.001) than on day 28 (P = 0.046) (None of the factors depicted in n day 28  and arou= 0.046) . Within-= 0.046) . Higher = 0.046) . They we= 0.046) . No asso= 0.046) . There wP < 0.001) (P < 0.001) (P = 0.070) with protoscolex motility, which, however, was abolished with adjustment for cyst concentrations (not shown). No other trend of univariate association between plasma concentrations and parasitological outcomes was observed. None of the factors depicted in P < 0.250) toward univariate association with either parasitological outcome (not shown). With adjustment for age, sex, “fertile cyst,” and obtaining of sample during surgery or PAIR, ASO cyst concentrations (by 0.01 mg/L) were independently associated with lower odds of motile protoscolices  and higher odds of “good or very good” morphological response .For all patients, parasitological findings were identical after 24 hours and after 48 hours of incubation with canine bile/sodium taurocholate. Data for 48 hours are presented. Cyst ASO concentrations were strongly univariately associated with parasitological outcomes: 0.01 mg/L increase in concentration was associated with a 39% reduction in odds of finding signs of protoscolex motility (< 0.001)  and with< 0.001) . Cyst AS< 0.001) . Plasma P = 0.035) or only surgically treated patients (P = 0.041) , all with previous (before the index) disease episodes and all tested seropositive at the start of the index treatment. Their characteristics were: case 1) age 66 years, CL (fertile) cyst, ASO plasma concentrations 0.149 and 0.132 mg/L on days 10 and 28, respectively, and cyst concentration 0.502 mg/L. Parasitological findings indicated no signs of protoscolex motility and >66% protoscolices with signs of destruction; case 2) age 42 years, CE1 (fertile) cyst, ASO plasma concentrations 0.290 and 0.188 mg/L on days 10 and 28, respectively, and cyst concentration 0.222 mg/L. Parasitological finding indicated protoscolex motility, and 33%-66% protoscolices with signs of destruction; case 3) age 72 years, CE3 (signs of involution) cyst, ASO plasma concentrations 0.188 and 0.042 mg/L on days 10 and 28, respectively, and cyst concentration 0.155 mg/L. Parasitological findings indicated no signs of protoscolex motility and <33% protoscolices with signs of destruction. With adjustment for age and presence of protoscolex motility, higher day 10 plasma ASO concentrations were associated with around 50% lower odds of disease recurrence considering all (= 0.041) . Day 28 = 0.041) , but not= 0.041) .The present study emphasizes the importance of attaining higher concentrations of the active albendazole metabolite, ASO, in the hydatid cyst in order to achieve a more pronounced protoscolex inactivation in patients with liver hydatidosis. The study further suggests two important conclusions: a) that cyst concentrations might not be predictable from the plasma concentrations; and b) that neither ASO concentrations in the cyst nor the level of protoscolex viability  are predictive for a long-term clinical outcome. Rather, data suggest that higher plasma ASO concentrations (reflecting the tissue concentrations) are associated with a reduced risk or disease recurrence. Introduction of albendazole in treatment of hydatid cyst disease enabled a more liberal surgical approach and development of percutaneous cyst evacuation techniques by reducing the risks associated with spillage of the cyst content -8. AttemThe limited number of patients was due to a relatively low disease incidence and our intention not to exceed a 1.5-year recruitment period in order to maintain consistency of study procedures. In this respect, and considering that the disease relapsed in only 3 patients during the study follow-up, the present observations relating higher plasma ASO concentrations to a reduced risk of disease recurrence should be taken with caution. It should be noted, however, that our intention was not to detect “all possible factors” that might have influenced any of the analyzed outcomes, but to test the pre-defined hypotheses about ASO concentrations while accounting for (likely) the most relevant confounders. In line with this, the fact that we found no relationship between patient or disease characteristics  with plasma or cyst ASO concentrations, parasitological outcomes, or disease recurrence does not “disqualify” these factors as potentially relevant – the study was simply not designed to formally test their effects. At the same time, this did not compromise our observations. On the other hand, lack of drop-outs, high adherence to the procedures, the use of a validated method of ASO quantification, and a standardized parasitological evaluation enabled a fair level of internal validity. Indirectly, present observations were externally validated by the fact that the observed ASO cyst concentrations were within the range of values reported in other studies with a similar dosing schedule, which, however, were conducted in considerably fewer patients ,18,19. Tmax, after the first morning dose), the observations related to plasma ASO concentrations were in agreement with the known high inter- and intra-subject variability. The fact that the values on day 10 of treatment were higher than on day 28 (end of treatment) could be due to several reasons. In a pharmacokinetic study in healthy volunteers , it seems pharmacodynamically plausible. The risk of disease recurrence after surgical cyst removal/PAIR is largely attributed to contamination due to the spillage of the cyst content. Clearly, the ability of shed protoscolices to generate a new cyst will depend on their viability. However, tissue ASO concentrations are also relevant since they could inhibit otherwise viable protoscolices. Animal studies show that ASO is readily distributed into different tissues where attained concentrations are comparable to plasma concentrations  or are two  to four times higher (liver)  . Once cymax on days 10 and 28 of treatment. However, this does not exclude the possibility that a more detailed plasma concentration-time profiling would be predictive of the cyst ASO concentrations. At the same time, the study strongly suggests that higher plasma concentrations are associated with a lower risk of disease recurrence.In conclusion, the present analysis in patients with liver hydatidosis treated with albendazole 15 mg/kg/d over 28 days before surgery or PAIR demonstrates the importance of attaining higher ASO concentrations in the cyst for reduction of protoscolex viability. Cyst concentrations were not predictable from plasma concentrations determined at the expected morning C


Previous studies have characterized an increasing trend of double burden households, or households with individuals experiencing both undernutrition and obesity, in countries undergoing a nutrition transition. Although most prior studies indicate the prevalence of double burden households is highest in middle-income countries, there is some support for an increase in double burden households in sub-Saharan African countries as well.Using data from the Demographic Health Surveys (DHS) and the World Health Organization (WHO), the prevalence of double burden households in sub-Saharan African countries was calculated and the associations between prevalence of overweight/obese adults and underweight, stunted and wasted children were evaluated at the country and household (DHS only) levels. Restricted analyses and frequencies were calculated using urban-only datasets. Surveys from 28 African countries were available using WHO data and 26 from the DHS surveys. Only surveys that were conducted after 2000 were included in analyses.Using the WHO datasets, there were inverse associations between the prevalence of overweight and obesity in adults and underweight, stunting and wasting in children. Correspondingly, there were positive associations between adult underweight and child underweight, stunting and wasting. These associations were not significant in a smaller sample size using urban-only surveys. The prevalence of double burden households in DHS datasets was low: under 5 percent for obese mothers and underweight, stunted or wasted child pairs with a slightly higher percentage for overweight mothers and children with undernutrition. Restricting the analysis to urban only populations did not increase the frequencies of double burden households significantly.There was a low prevalence of double burden households in recent data from sub-Saharan Africa. Countries that have a high prevalence of child undernutrition correspondingly have a high prevalence of adult underweight and low prevalence of adult overweight and obesity. The number of overweight and obese women has continued to increase internationally including in low-income countries such as those in sub-Saharan Africa . MeanwhiRecent studies have found an increasing trend of ‘dual burden’ or double burden households, households that have both underweight or stunted and overweight/obese persons , 8. In pIn a report of data from the Demographic and Health surveys of 23 sub-Saharan African countries, African countries surveyed had less than 10% of households with stunted child-overweight mother pairs with the exception of Egypt, which had 14% . A studyIn a more recent study using Demographic Health Survey data from the early 2000s from North Africa, sub-Saharan Africa, Asia and Latin America, Jehn and Brewis  found thIn this review of double burden households in sub-Saharan Africa, we assess both the association between maternal body mass index (BMI) category and child undernutrition using country level data  and the prevalence of double burden households using household level data as described below (DHS data). In contrast with previous studies, we evaluate associations using post-2000 data, with a particular emphasis on data collected after 2005.Two different survey sets were used to evaluate the prevalence of double burden households and estimate the possible association between maternal overweight/obesity and child undernutrition. These datasets were publically available and no unique identifiers were available. The WHO surveys were included in this analysis given the large number of surveys conducted in each country. However, as the data were not collected at the household level, it was not possible to assess the prevalence of double burden households but only evaluate the country-level associations between adult BMI category and child undernutrition. For WHO data, in some case women and in other cases combined women and men data are available. The Demographic Health Surveys were also included in this study so as to be able to assess the prevalence of double burden households in sub-Saharan African countries using household level data, albeit using a smaller number of surveys per country than those available in the WHO database. The specific datasets and methods used are described in detail below.2), overweight (BMI ≥25 m/kg2), underweight (BMI <18.5 m/kg2) and child stunting (having a height-for-age Z score < -2 standard deviations (SD) below the international reference median or < -3SD for severe stunting, underweight , overweight (having a BMI-for-age z-score > +2 SD) and obesity (BMI-for-age z score > +3 SD). Data from the following 28 sub-Saharan Africa countries were included from the WHO databases: Benin, Burkina Faso, Cameroon, Chad, Congo, Democratic Republic of the Congo, Eritrea, Ethiopia, Gabon, Ghana, Kenya, Lesotho, Liberia, Madagascar, Malawi, Mali, Mauritania, Morocco, Mozambique, Namibia, Niger, Nigeria, Rwanda, Sierra Leone, Swaziland, Uganda, Zambia, Zimbabwe. WHO compiles data collected from national and sub-national surveys [To estimate the possible relationship between maternal overweight/obesity and child stunting and underweight, we used the World Health Organization’s (WHO) Global Database on Child Growth and Malnutrition and the WHO’s Global Database on Body Mass Index to assess a possible correlation between frequencies of adult obesity (a body mass index (BMI) ≥30 m/kg surveys , 12. We  surveys , 12. ChiThe WHO Global Database on Body Mass Index combined BMI results from men and women and included the use of 3,169 surveys and smaller studies from 28 countries . The criteria WHO uses for surveys included in its database is a defined population-based sampling frame and a probabilistic sampling procedure involving at least 400 children . Using tFurthermore, we conducted a smaller subset analysis of urban surveys only with data from 23 countries, which included 89 surveys of which 58 were women only. Means and standard deviations were calculated for each country for percentage of BMI in the overweight and/or obese category and child anthropometrics using Stata 13.0. Linear regression was subsequently used to evaluate relationships between mean frequencies of adult overweight/obesity and mean levels of stunting, wasting and underweight by country.Sampling weights for the different surveys were not provided by WHO. Subsequently, analyses were restricted to adult female BMI numbers only and then to surveys conducted only among urban residents. In the analyses that included all adults and women only, the sample size was n = 28 African countries for both adults and only women while in the urban only analyses the sample size was n = 23 African countries.The WHO surveys include surveys from UNICEF, UN Statistics Division, FAO, the World Bank, International Food Policy Research Institute (IFPRI) and the Demographic and Health Surveys (DHS) among others . We usedAnalysis was conducted also using the Demographic Health Surveys (DHS) for the following 26 sub-Saharan African countries specific years: Benin (2001), Burkina Faso (2003), Cameroon (2004), Congo (Brazzaville) 2005, Gabon (2000), Ghana , Guinea (2003), Kenya , Lesotho , Liberia (2007), Madagascar (2003), Malawi (2000), Mali , Mozambique (2003), Namibia (2007), Niger (2008), Nigeria , Rwanda (2000), Sao Tome (2009), Senegal (2005), Sierra Leone (2006), Swaziland (2006), Tanzania , Uganda (2006), Zambia , Zimbabwe (2005). Countries were not included if they did not have a survey conducted in 2000 or later or if they did not include a measurement of maternal BMI and child anthropometrics as part of the information collected.2, overweight (BMI ≥ 25 m/kg2), underweight (BMI <18.5 m/kg2) and child stunting (having a height-for-age Z score < -2 standard deviations (SD) below the international reference median), underweight (having a weight-for-age Z score < -2 SD) and wasting (having a weight-for-height Z score < -2SD). Analyses of double burden household prevalences used the definitions of undernutrition  given the low prevalence of double burden households using the more expansive definition. All analyses were conducted using Stata 13.0 and with svy commands.The children’s file which included mother’s BMI measurement and child anthropometrics was used for all analyses. Sampling weights were used in calculations as provided by the DHS . In the The mean prevalence of adult obesity in the WHO dataset was 7.5 ± 6.0% while adult overweight was much higher at 21.8 ± 10.2% and adult underweight was 13.4 ± 7.0%. Stunting (<-2SD) was 38.7 ± 7.8% and severe stunting (<-3SD) was 19.0 ± 5.7%. Underweight (<-2SD) was 21.5 ± 8.8% and severe underweight (<-3SD) was 7.96 ± 4.3%. Wasting (<-2SD) was 10.0 ± 4.4% and severe wasting (<-3SD) was 4.0 ± 2.2%. In the urban surveys, the prevalence of adult obesity and overweight were higher (12.5 ± 7.0% and 31.8 ± 12.8%), child undernutrition was generally lower and adult underweight (10.5%) was also lower.In both the overall combined analysis using data from men and women and the analysis of just women, we found that there were significant relationships between the prevalence of adult BMI category and % of children underweight, wasted, stunted, overweight or obese Tables , 3 and 4Specifically, looking at the associations in adult men and women, wasting (<-3SD) was positively associated with adult underweight, while wasting (<-2SD) was positively associated with adult underweight and negatively associated with adult overweight and obesity Table . For stuIn the restricted analyses, adult women only, the results were comparable to those that included men and women Table . Using aThe associations between adult obesity and underweight, and child severe stunting and severe underweight, are also shown graphically in Figures The overall prevalence of maternal overweight and obesity in the 27 DHS countries surveyed was 16.5% (95% CI 16.1-17.0%), maternal obesity was 5.1% 95% CI 4.8-5.3%) and maternal underweight was 10.4% (95% CI 10.1-10.7%). The prevalence of maternal overweight and obesity was higher than 20% in the following countries and years: Congo 2005, Ghana 2008, Namibia 2007, Nigeria 2003, Nigeria 2008, Sao Tome 2009, Sierra Leone 2006, Swaziland 2006 and Zimbabwe 2005  was less than 5% for all the countries surveyed with the exception of Swaziland in 2006, which had a prevalence of 9.50% for the pairing of an obese mother with an underweight child , 1.79% (95% CI 1.62-1.98%) for obese mothers/stunted child pairs and 0.50% (95% CI 0.42-0.59%) for obese mothers/wasted child pairs , 4.74% (95% CI 4.56-4.92%) for underweight mother with stunted child, and 1.50% (95% CI 1.41-1.59%) for underweight mother with wasted child , a clear trend in the association between childhood undernutrition and maternal obesity or overweight does not emerge.The overall prevalence of double burden households in the sub-Saharan African countries surveyed was not significantly higher than that found by the other studies, , 9 whichWhen the analyses were restricted to urban areas or women with a higher educational background, the prevalence of double burden households did not change remarkably, although slightly higher for maternal obesity and child undernutrition households. This was not surprising as the overall percentage of undernutrition in children is lower in urban areas compared with national prevalences even though the prevalence of obese and overweight women is higher. Other studies using African DHS data to evaluate trends over time have found that the prevalence of urban adult overweight and obesity increased by 35% from 1992 to 2005 approaching 30% in urban areas  and undeThese findings are in contrast with some previous studies, which have found a higher prevalence of double burden households in sub-Saharan African countries and have predicted an increase in prevalence in coming years such as the study by Bouzitou et al. in 2005,  which foJehn and Brewis  concludeWe also found a high prevalence of overweight and obesity in women in all the sub-Saharan African countries surveyed, particularly in urban areas, in excess of 30% in many sub-Saharan African countries and exceeding 40% in a few (Figure In the regression analyses using WHO datasets which included more surveys than the DHS analysis, there was a negative association between overweight/obesity in women and child undernutrition and a positive association between underweight in women and child undernutrition as expected. We did not see any negative relationship between overweight/obesity in women and child undernutrition when restricting the analysis to urban areas only. It is possible that because of the relatively high overall prevalence of adult overweight/obesity irrespective of household category, particularly in the urban only surveys, greater power is needed to assess any statistical association between overweight/obese mothers and children with undernutrition.In the WHO and DHS analyses, there was no adjustment or stratification based on household income levels. Previous studies have found that the highest income households, particularly in urban areas were the most likely to be double burden households with the study by Doak et al.  reportinAs argued by Garrett and Ruel , it is pFuture studies need to assess the dietary intake of double burden households in contrast with normal weight or underweight households. In the study by Bouzitou et al.  from BenFurther studies need to be conducted on the relation between household income and risk for double burden households in sub-Saharan African countries as well as to better characterize the nutrition transition in African countries. These studies should focus on explaining the seeming paradox of a low prevalence of double burden households in urban areas with increasing adult obesity rates. Furthermore, the increasing obesity prevalence and the failure to decrease child under-nutrition in African countries suggested by Ziraba et al.  and de O


Incidence rates of various cancers are increasing in Arab countries and are expected to reach those of industrialized ones in few decades. This paper aimed to describe the incidence rates of most common cancers - and/or of those cancer preventable through modifiable behaviors - recorded in the province of Setif, Algeria from 1986 through 2010.Cancer diagnoses for the 1986–2010 period were provided by the population-based Cancer Registry of Setif, disentangled by site, morphology, age (quinquennia), sex, and calendar period. The corresponding population was obtained from the Algerian Institute of Statistics. Age-standardized rates (world population) (ASR-WR) were computed by calendar period (five quinquennias from 1986–1990 to 2006–2010), while annual percent changes (APCs) were computed for the period 1996–2010.During the 2006–2010 period, ASR-WR for all cancer sites were 106.4/100,000 in men and 110.3 in women. The four leading cancers were: lung (18.0%); colon-rectum (9.6%); bladder (9.1%); and prostate (6.5%) in men; breast (36.4%); colon-rectum (8.5%); cervix uteri (6.0%); and thyroid (6.0%) in women. Between 1996–2010, overall cancer incidence increased statistically significantly (p < 0.05) in both men (APC = +2.5%) and women (APC = +3.7%). Statistically significant decreasing trends were observed for nasopharyngeal carcinoma (APC = -3.4%) in men, and for cervical (APC = -4.2%) and gallbladder (APC = -3.2%) cancers in women. Statistically significant increasing trends were observed for most common cancers both in men  and women .International recommendations against cancer must be strongly promoted in Setif after taking into account epidemiological transition, lifestyle, and environmental changes. Cancer rates in northern Arab countries lying on the African side of the Mediterranean Sea are substantially lower than those reported in European Mediterranean countries such as Spain, France or Italy . HoweverThe cancer registry of Setif (CR-S), Algeria, has been recording incidence data for the whole Wilaya (province) of Setif, the second in size after the Wilaya of Algiers - the capital city of Algeria. An accurate description of temporal trends by cancer site can contribute to a better understanding of cancer incidence patterns and their potential etiology and/or prevention. We took advantage of the availability of computerized incidence data recorded in CR-S over the 1986–2010 period to describe trends in incidence for the most common cancer sites and/or for the potentially preventable ones.Data regarding all incident cancer cases diagnosed during the period 1986–2010 derived from the population-based cancer registry of Setif, established in 1989 in collaboration with the International Agency for Research on Cancer (IARC). The Director of the Cancer Registry of Setif (Prof. Hamdi Cherif) approved the use of the registry data for the purposes of the study. Nowadays, the CR-S covers a population of about 1.5 million inhabitants , which is nearly 5% of the Algerian population. Collection of data is primarily active.For the aims of this analysis, incidence data were disentangled by age (quinquennia), sex and type of cancer. The International Classification of Diseases for Oncology, second version (ICD-O-3), was used for cancer classification . The ICDhttp://www.ons.dz].The structure of the resident population by sex, age, and calendar year was abstracted from the Algerian Institute of Statistics database [Incidence data were grouped into five quinquennia of calendar years . Age-standardized incidence rates per 100,000 (to the world standard population) [The computation of annual percent change (APC) , 12 of iThe overall number of incident cancers  diagnosed in the Wilaya of Setif during the period 2006–2010 was 6235 (2883 men and 3352 women) corresponding to an ASR-WR  of 106.4 in men and 110.3 in women (Table Throughout the study period 25 years), overall cancer incidence rates steadily increased in both sexes Table . In men,5 years, In men, stomach cancer incidence rates decreased continuously during the whole period. Nasopharyngeal incidence rates increased up to 1996–2000 and declined thereafter. Non-Hodgkin’s lymphomas rates increased up to 1996–2000 and leveled-off thereafter. Cancers of gallbladder and kidney, Hodgkin lymphomas, leukemias and ill-defined cancer showed somewhat stable trends during the 25 years examined. Liver cancer rates increased since 1996–2000 after a decrease. Conversely, increased trends were observed for cancer of the colon-rectum, larynx, lung, prostate, bladder and thyroid (since 1996–2000).In women, stomach cancer rates declined up to 1996–2000 and leveled-off thereafter, whereas rates of invasive cervical cancer increased up to 1991–1995 and declined thereafter. Gallbladder rates increased up to 2001–2005 and declined thereafter while lung cancer incidence rates increased after 2001–2005. Cancers of nasopharynx, larynx, ovary, kidney, and bladder, Hodgkin’s lymphomas, leukemias and ill-defined cancers showed somewhat stable trends during the 25 years examined. Liver cancer rates increased since 1996–2000 after a decrease. By contrast, cancer of the colon-rectum, breast, and thyroid, and non-Hodgkin’s lymphomas increased throughout the whole period examined.Furthermore, between 1996 and 2010, cancer incidence rates showed some statistically significant negative APCs (% per year) , and stomach cancer (APC = -7.0) in men, and for stomach (APC = -6.6), and gallbladder (APC = -8.7) cancers in women. Increases were observed for leukemias (APC = +13.7) in men, for cancers of the breast (APC = +9.7), and thyroid (APC = +9.8) and non-Hodgkin lymphomas (APC = +6.3), and leukemias (APC = +11.2) in women. The increase of leukemias incidence was mainly related to the increase of lymphoid leukemias, however no complete information was available about the type of lymphoid leukemia . The 45–64 age group displayed statistically significant decreases for nasopharynx carcinoma (APC = -4.0), in men, and for cervical (APC = -5.9) and ovarian (APC = -7.5) cancers in women. Increases were observed for colon-rectum (APC = +3.7), and bladder (APC = +3.5) cancers in men, and for colon-rectum (APC = +5.3), lung (APC = +10.4), breast (APC = +7.3), and thyroid (APC = +5.0) cancers in women. The 65+ age group displayed exclusively statistically significant positive APCs for colon-rectum (APC = +11.1), lung (APC = +2.8), prostate (APC = +5.0), and bladder (APC = +8.0) cancers in men, and for colon-rectum (APC = +7.5), lung (APC = +7.8), and breast (APC = +6.9) cancers in women.We investigated cancer incidence trends recorded by the Cancer Registry of the Wilaya of Setif, Algeria, across a 25 year period (1986–2010). To the best of our knowledge, this is the first report to investigate a large number of different types of cancer according to age for a quarter century in a northern African population. Decreasing trends were observed for nasopharyngeal carcinoma in men, and for cervical and gallbladder cancers in women, whereas increasing ones were observed mainly in leading cancers both in men  and women .Leading cancers were similar to those reported by other North-African studies , 13, 14.Lung cancer incidence is largely determined by prevalence and duration of smoking. In Algeria, between 1970 and 1980, prevalence of smoking increased both for blond 34 times) and for black (+50%) tobacco. In addition between 1978 and 2005 prevalence of smoking increased from 7.7% to 28.6% 4 times a, 16. SmoNasopharyngeal carcinoma (NPC) is a rare malignancy in most regions of the world, but it is endemic in Algeria and more generally in North Africa. Major risk factors include Epstein-Barr virus infection, genetic susceptibility, consumption of salt-preserved fish or other preserved foods, a family history of NPC, and tobacco smoking . In SetiChanges in viral hepatitis prevalence rather than alcohol consumption seemed to explain the rise of liver cancer incidence because the other alcohol-related cancers  were stable, with ASR-WRs below 1/100.000 (not shown). The prevalence of hepatitis B virus in blood donors was around 3.6% in 1989 [Breast cancer, the most common female cancer diagnosed in Setif, dropped in all age groups. The reasons for this shift are unknown, but they may include increases in the prevalence of risk factors that modify endogenous levels of sex hormones such as early menarche, late menopause, late childbearing, having fewer children, and low prevalence of breastfeeding, and unhealthy diets, physical inactivity or obesity , 22. ForInvasive cervical cancer (ICC) is the third most common female cancer in Setif, and a sharp decline in ICC incidence was observed mainly in the 15–44 age group. A potential shift of ICC diagnoses to uterus, unspecified  cannot explain this decline. The development of cervical cancer is causally associated to infection by high-risk Human Papilloma Viruses (HPV). In Algeria, the HPV16 in single- or multiple-type infection has been shown to be the predominant HPV type (61.4%) in ICC, followed by HPV18 (15.8%), and HPV45 (6.4%) . The preThyroid cancer (TC) incidence increased in women in 15–44 and 45–64 age groups. In 2006–2010, the most common form of TC was the papillary sub-type and was related to most of the increased incidence . This is thought to be largely due to increased detection of small papillary cancers associated to widespread use of ultrasound and fine needle biopsies .Prostate cancer is the third most common cancer in men in Algeria. A number of risk factors detected in Western Algeria can explain the increased trend in prostate cancer incidence, such as high consumption of red meat, and dairy products, or low consumption of olive oil, cauliflower, and intake of fruits . PresentReduction of stomach cancer incidence is generally attributed to diets rich in fruit and vegetables, improved preservation of foods, and reduced prevalence of Helicobacter pylori . Some dind most common cancer in both sexes. The adoption of western lifestyles and behaviors such as consumption of high-fat diets and physical inactivity could explain the increase of these cancers together with genetic predisposition [Colon-rectum cancer increased in 45–64 and 65+ age groups and is the 2position . It is pth most common cancer in Setif women. Algerian women show the highest rates of gallbladder cancer as compared to the other Arab countries where this cancer is generally rare [Gallbladder is the 5lly rare . The temThe selective increase of non-Hodgkin’s lymphomas in the 15–44 age group in women is intriguing. Moreover, further studies are required to elucidate the increase of leukemias, particularly lymphoid leukemias, in the 15–44 age group.Incidence data have potential limitations in accuracy and completeness that should be made clear. Firstly, cancer and comorbidities are extremely common in elderly people and the percentage of histologically confirmed cancers is generally lower among them than in younger age groups. Although under-diagnosis and under-registration seem to be unlikely, the complete identification of true cases in older age groups may be difficult. Secondly, the accuracy of DCOs depends on coding practices. The low number of DCO should have limited the possibility of differential practices influencing death certification procedures. Thirdly, no mortality/incidence ratios were available . This information would have given further confidence in the observed results. Fourthly, due to the relatively small population covered , more accurate results from a bigger cohort are needed.Among the strengths of the study, MV% reached good or acceptable levels in the studied cancer sites. Moreover MV% did not change significantly over time, thus, any estimated trend in incidence is unlikely to be the consequence of changes in the degree of ascertainment. Moreover, a small proportion of cancer cases were reported by DCOs. Finally, the availability of complete morphology data provided the opportunity to associate some trends to potential early detection initiatives.Some main conclusions can be drawn from the trends of cancer incidence observed in the last 25 years in Setif. Besides ageing of the population, temporal variations in caseload indicate cohort effects due to differential exposure to risk factors and improved detection methodology. In the light of international recommendations against cancer  three le


A population PK model was constructed, and the probability of target attainment (PTA) for 50% and 100% fT>MIC was calculated for varying MIC and CLCR values.To be eligible for enrolment, critically ill patients with sepsis had to be receiving piperacillin-tazobactam 4.5 g intravenously (IV) by intermittent infusion every 6 hours for presumed or confirmed nosocomial infection without significant renal impairment . Over a single dosing interval, blood samples were drawn to determine unbound plasma piperacillin concentrations. Renal function was assessed by measuring creatinine clearance , such that with an MIC of 16 mg/L, 100% fT>MIC would be achieved in only one-third (n = 16) of patients. Mean piperacillin clearance was approximately 1.5-fold higher than in healthy volunteers and correlated with CLCR . A reduced PTA for all MIC values, when targeting either 50% or 100% fT>MIC, was noted with increasing CLCR measures.In total, 48 patients provided data. Increasing CLCR can be employed as a useful tool to determine whether piperacillin PK/PD target attainment is likely with a range of MIC values.Standard intermittent piperacillin-tazobactam dosing is unlikely to achieve optimal piperacillin exposures in a significant proportion of critically ill patients with sepsis, owing to elevated drug clearance. These data suggest that CL Effective antibacterial therapy is crucial for improving outcomes for patients with sepsis . CurrentIn vivo animal data indicate that this period should be at least 40% to 70% of the dosing interval (40% to 70% fT>MIC) [fT>MIC [min:MIC) >5 [fT>MIC [The minimum antibacterial exposure required in clinical practice to maximize bacterial killing and optimize clinical outcomes remains controversial. For β-lactams, preclinical studies support maintaining prolonged free drug concentrations above the minimum inhibitory concentration (MIC) of the likely pathogen . In vivo [fT>MIC  and trou:MIC) >5 . More reWhereas these data demonstrate why achievement of target drug exposures is important, they do not robustly define which patients are at risk of subtherapeutic dosing. Clinicians commonly consider the likely pathogen, its susceptibility profile and the potential for drug toxicity when selecting an empirical dosing regimen. An assessment of renal function is common, with evidence of acute or chronic renal impairment often triggering dose reduction to avoid drug accumulation. For β-lactams, the converse—dose escalation in the setting of augmented renal function—has been infrequently reported in clinical practice.CR), are at particular risk of subtherapeutic β-lactam concentrations [CR, pathogen susceptibility and drug exposures. As such, the aim of this analysis was to explore the impact of elevated CLCR and different pathogen susceptibilities, on piperacillin pharmacokinetic/pharmacodynamic (PK/PD) target attainment in critically ill patients receiving intermittent dosing.Previously published data, however, suggest that patients with augmented renal clearance (ARC), which often manifests as an elevated urinary creatinine clearance . Ethical approval was obtained from the Royal Brisbane and Women’s Hospital Human Research Ethics Committee (HREC 2007/188), with written informed consent obtained from either the patient or the patient’s nominated substitute decision-maker.Patients were eligible for enrolment if they were between 18 and 80 years of age and were receiving piperacillin-tazobactam for the treatment of sepsis . PatienCR. No specific CLCR value was used to define renal impairment or ARC, as our aim was to use these measures as continuous variables in analysis.The protocol pertaining to this study has been published in detail elsewhere . In brieAdditional data, including the requirement for mechanical ventilation, vasopressor support, modified Sequential Organ Failure Assessment (SOFA) score  and 24-hour fluid balance, were documented on the day of drug administration. Admission Acute Physiology and Chronic Health Evaluation (APACHE) II score, body mass index (BMI) and ICU and in-hospital mortality were also recorded.Blood samples were immediately placed on ice and centrifuged within 60 minutes at 3,000 rpm for 10 minutes. Plasma samples were then stored at −80°C until analysis. An ultraviolet high-performance liquid chromatography (HPLC-UV) assay was used to measure piperacillin concentrations in plasma. All bioanalytical techniques were validated and conducted in accordance with the criteria of the US Food and Drug Administration’s guidance for industry on bioanalysis . To isolmin). These samples were then compared to an MIC of 16 mg/L to determine whether 100% fT>MIC would have been achieved. This represents the highest MIC for susceptible bacteria  as per the European Committee on Antimicrobial Susceptibility Testing (EUCAST) [fT>MIC target was utilized in this analysis, as more aggressive drug exposures have been demonstrated to improve both microbiological and clinical outcomes [The final (360 minutes) plasma sample was taken as the trough unbound piperacillin concentration (C(EUCAST) . We chosoutcomes ,8,9.Concentration-time data were analysed using nonlinear mixed-effects modelling . A Digital Fortran compiler was used, and the runs were executed using Wings for NONMEM . The fir2-distributed. On the basis of a χ2 test of the difference in OFV, a decrease in the OFV of 3.84 units (P < 0.05) for 1 degree of freedom was considered statistically significant. Decreases in BSV of one of the parameters of at least 10% were also accepted for inclusion in a more complicated model.Visual inspection of diagnostic scatter plots and the NONMEM objective function value (OFV) were used to evaluate goodness of fit. Statistical comparison of nested models was undertaken in the NONMEM program using log-likelihood ratios, which are assumed to be χn = 1,000) was used to study the uncertainty of the pharmacokinetic parameter estimates in the final model. From the bootstrap empirical posterior distribution, we were able to obtain the 95% confidence interval (2.5% to 97.5% percentiles) for the parameters, as described previously [A nonparametric bootstrap method (eviously .CR (10 to 300 ml/min in 10-ml/min increments) values using a standard 4.5-g piperacillin-tazobactam dose administered every 6 hours as an intermittent 20-minute infusion. The PTAs for achieving 50% fT>MIC and 100% fT>MIC with piperacillin were calculated.Monte Carlo simulations performed in NONMEM were employed to determine the probability of target attainment (PTA) with varying MIC (2 to 64 mg/L) and CLPseudomonas aeruginosa against piperacillin was obtained from the EUCAST database [fT>MIC and 100% fT>MIC by using a range of CLCR values. Dosing was a priori considered successful if the CFR was at least 80%.The cumulative fraction of response (CFR) determines the likely success of treatment by comparing the pharmacodynamic exposure (PTA) against the MIC distribution of a specific population of microorganisms . The wildatabase . The CFRr). For comparisons between groups, we used an independent Student’s t-test, Mann–Whitney U test, or one-way analysis of variance for continuous data, as well as a χ2 or Fisher’s exact test for categorical data, where analysis assumptions were met. A P-value <0.05 was considered statistically significant, and all analyses were performed using SPSS version 21 software .Continuous data are presented as the mean (standard deviation) or median (interquartile range). Categorical data are presented as counts (%). Correlation was assessed by means of a scatter graph and Pearson correlation coefficient (CR over the study period was 122 (59.2) ml/min, although there was significant variability between patients: 15 patients had a CLCR <90 ml/min, 22 had a CLCR >130 ml/min and 16 had a CLCR >150 ml/min.Forty-eight patients were included in the analysis. All patients were receiving 4.5 g of piperacillin-tazobactam by intermittent IV infusion over 20 minutes every 6 hours for treatment of presumed or confirmed nosocomial infection. Demographic, admission, anthropometric, illness severity and outcome data are presented in Table min value was either below the lower limit of detection (n = 10) or not available (n = 1). Using an MIC of 16 mg/L, piperacillin intermittent infusion would achieve 100% fT>MIC (Cmin >16 mg/L) in only 16 (34.0%) of 47 patients. Older age (P = 0.01), higher modified SOFA score (P < 0.01), lower CLCR (P < 0.01), greater positive 24-hour fluid balance (P = 0.04) and more frequent application of vasopressor therapy (P = 0.01) were characteristics of patients likely to have higher Cmin values, and therefore achieve this PK/PD target. No statistically significant differences were noted in BMI, APACHE II scores or the need for mechanical ventilation. Unbound plasma piperacillin concentration time profiles grouped by CLCR quartile are presented in Figure CR was significantly associated with lower unbound trough plasma piperacillin concentration (P < 0.01).In 11 patients, the Cc) and peripheral compartments (Vp), drug clearance (CL) and an infusion lag descriptor . The only covariate that improved the fit of the model was, for piperacillin CL, CLCR normalized to 100 ml/min, which decreased the OFV by 18.62 (P < 0.01). In the final model, this was described as follows:The time course of unbound plasma piperacillin concentrations was best described by a two-compartment linear model with combined residual error and BSV on the volume of the central (Vd) was calculated as the sum of Vc and Vp. The goodness-of-fit plots were acceptable with an r-value of 0.95 for the observed versus individual predicted concentrations.where TVCL is the typical value of piperacillin CL. The mean parameter estimates from the final covariate model, as well as the 95% confidence intervals from all bootstrap runs, are shown in Table fT>MIC (16 mg/L) had higher drug CL (P < 0.01) and no statistically significant difference in piperacillin Vd (P = 0.647). As demonstrated in Figure CR and piperacillin CL overall . The PTAs for 50% fT>MIC and 100% fT>MIC with varying MIC and CLCR values are presented in Figure Those patients who would fail to achieve 100% CR values using the EUCAST MIC distribution for Pseudomonas aeruginosa is presented in Table The CFR for piperacillin over a range of CLCR and different susceptibilities of bacteria on piperacillin PK/PD target attainment when administered by intermittent infusion in a large cohort of critically ill patients with sepsis. With a MIC at the upper limit of the susceptible range for piperacillin (16 mg/L), only about one-third of patients would achieve 100% fT>MIC. The primary mechanism underlying this observation appears to be significantly elevated drug CL in parallel with higher CLCR in some patients. This phenomenon of augmented renal clearance [CR values (<90 ml/min) will an adequate fraction of Pseudomonas aeruginosa isolates be suitably covered with this dosing regimen.In this clinical analysis, we sought to explore the impact of elevated CLlearance ,20 is belearance ,22, whicd was also significantly larger overall. Changes in Vd have been observed with other β-lactams in critical illness [d observed, although use of free concentrations is essential to describe the pharmacologically active fraction of piperacillin.Pharmacokinetic modelling for the study cohort revealed a mean piperacillin CL of approximately 1.5-fold that reported in healthy volunteers . Similar illness  and like illness . Use of CR in patients with sepsis [et al. highlights the role of systemic inflammation, with an increasing number of systemic inflammatory response syndrome criteria being strongly correlated with higher drug clearance and lower plasma concentrations in nonventilated patients receiving vancomycin [CR or therapeutic drug monitoring should be employed to determine the need for higher than standard piperacillin dosing.The specific mechanisms underlying ARC in critically ill patients remain uncertain. Researchers have recently identified a modest correlation between cardiac index and CLh sepsis , suggesth sepsis ,29, whicncomycin . Togethencomycin . As suchCR in critically ill patients with ‘normal’ plasma CR concentrations, researchers identified an incidence of ARC (CLCR ≥130 ml/min/1.73 m2) of approximately 65% in the first 7 days in the ICU [The prevalence of ARC varies significantly, depending upon the definition employed and institutional case mix -35. In a the ICU . Importa the ICU .CR of 100 ml/min and a traditional regimen of 4.5 g administered IV every 6 hours, the probability of target attainment (50% fT>MIC) decreased sharply (<80%) with MIC values greater than 4 mg/L [CR thresholds , the probability of achieving either 50% fT>MIC or 100% fT>MIC was significantly reduced [CR as the only significant predictor of target attainment, a finding also demonstrated by Hites et al. in their work exploring β-lactam exposure in obese, non-critically ill patients [CR [More recently, Carlier  reduced . Regresspatients . Our datpatients . Dosing ents [CR .90 values for piperacillin-tazobactam against Pseudomonas aeruginosa were reported at or above 64 mg/L [fT>MIC in such a scenario is unlikely with conventional piperacillin intermittent dosing and reinforce the use of a much lower susceptibility breakpoint (16 mg/L) with this dosing strategy. However, even in this circumstance, an elevated CLCR will significantly reduce the probability of target attainment, as demonstrated in Figure CR values. Importantly, this underlines the significance of such microbiological susceptibility data in accurate clinical decision-making.Changing bacterial susceptibility represents a global challenge in medical practice. Our analyses highlight the importance of the MIC as the denominator in the PK/PD relationship in determining adequate antibacterial dosing. Less susceptible pathogens are more common in the ICU , and sig 64 mg/L ,46. Our CR) as a covariate in PK modelling. We did not include tazobactam PK, because it is not active against Pseudomonas aeruginosa, although, as it is renally eliminated, increased drug CL is also likely in ARC. We examined the likelihood of target attainment using varying susceptibility levels rather than clinical MIC data. This improves generalizability for empiric dosing, although it limits conclusions about the adequacy of drug exposure in any specific patient. Furthermore, given the small sample size, we are unable to make any firm conclusions about the clinical implications of these analyses.Our study has several limitations. The data were drawn from a single centre and as such may not be representative of case mix at other institutions. Our inclusion criteria were designed to select a cohort of patients without significant renal impairment, as effective antibacterial dosing in this group is less likely with standard dosing. As such, ARC is likely to have been more common in our study cohort than in a broader ICU population. However, our relatively high plasma CR cutoff allowed us to explore renal function , particularly with more resistant organisms.Increasing CLPseudomonas aeruginosa, 6-hourly dosing of 4.5 g piperacillin-tazobactam is unlikely to provide sufficient piperacillin exposure when the CLCR is ≥90 ml/min.When considering the MIC distribution of


The presence of drugs and their metabolites in surface waters and municipal effluents has been reported in several studies, but its impacts on aquatic organisms are not yet well understood. This study investigated the effects of acute exposure to the antipsychotic risperidone on the stress and behavioral responses in zebrafish. It became clear that intermediate concentration of risperidone inhibited the hypothalamic-pituitary-interrenal axis and displayed anxiolytic-like effects in zebrafish. The data presented here suggest that the presence of this antipsychotic in aquatic environments can alter neuroendocrine and behavior profiles in zebrafish. This drug is widely used for the treatment of schizophrenia and bipolar disorder. In people with schizophrenia in maintenance treatment, risperidone induces relatively few extrapyramidal syndromes (EPS), especially less akathisia and tremor, as compared to typical antipsychotic haloperidol. As expected, due to its high prescription rates and off-label uses [The contamination of water resources like natural water bodies or urban effluents by pharmaceutical drugs and/or its metabolites has been reported since the 1970’s, consequently increasing the concern with health consequences for human population as well as for the aquatic life [bel uses –13, rispThe activation of the hypothalamus-pituitary-interrenal (HPI) axis and the consequent cortisol elevation is an important reaction of the organism against physical stressors, predator encounters, and chemical stressors such as drug contaminations, aiming to restore the homeostasis –18 with Danio rerio) as the experimental model, since this species presents many advantages such as easy handling and maintenance as well as shows high genetic homology with humans [We hypothesized that risperidone residues, due its central effects, might alter the functioning of the stress neuroendocrine axis, impairing the general response of the fish residing in contaminated water bodies. We tested our hypothesis using zebrafish  and was approved by the Ethics Commission for Animal Use (CEUA) at Universidade de Passo Fundo, UPF, Passo Fundo, RS, Brazil (Protocol # 7/2013 –CEUA). The use of this method was justified to avoid potential confounding factors while interpreting data regarding the effects of risperidone on the stress response axis.Danio rerio) short-fin (SF) strain, weighing 0.7 to 0.9 g were held in glass aquaria with constant aeration and equipped with biological filtering under a natural photoperiod (approximately 14 h light: 10 h dark). Water was maintained at 26 ± 2°C and pH 7.0, with dissolved oxygen levels at 6 ± 0.5 mg/L, total ammonia levels at 0.01 mg/L, total hardness at 6 mg/L, and alkalinity at 22 mg/L CaCO3. Behavioral testing was performed during the afternoon.A population of approximately 500 mixed-sex, adult wild-type zebrafish  plus the controls (with or without stress). Thus, we have 12 glass aquaria with six fish each. We repeated this setup 3 times, using two fish from each aquaria in each replication, totalizing a final sample of 6 fish (n = 6).Zebrafish were distributed in 36 glass aquaria , acclimatized for seven days and fed with commercial food flakes . Twenty-four hours later, fish were exposed to risperidone for 15 minutes. Fish were then stressed by chasing them with a net for two minutes , and samRisperidone  was used in five concentrations: 0.00034 μg/L, 85 μg/L, 170 μg/L, 340 μg/L and 680 μg/L. The lower concentration was already detected in the environment . The envWe performed a behavioral test  usiFish were captured and immediately frozen in liquid nitrogen for 10–30 s, followed by storage at -20°C until cortisol extraction. Whole-body cortisol was extracted using the method described by Oliveira et al. . Tissue Whole-body cortisol levels were analyzed by three-way Analysis of Variance (ANOVA) with treatment, stress and sampling time as the independent variables, followed by two-way ANOVAs restricted to each stress condition. Differences at each time point were analyzed by Bonferroni post-hoc test. Behavioral parameters were analyzed by two-way ANOVAs with treatment and stress as the independent variables, followed by Tukey’s multiple range post-hoc test. For both data sets, the homogeneity of variance was determined using Hartley’s test, and normality was determined using the Kolmogorov-Smirnov test. Data were expressed as means ± standard error of mean (S.E.M). Significance level was set at p<0.05.The acute stress protocol increased cortisol levels in zebrafish. There was a significant interaction among treatment, stress and time. Zebrafish exposed to 170 mg/L of risperidone and submitted to the acute stress test had a reduced cortisol response to an acute stressor at 15, 60 and 240 minutes after stress . SignifiThe Here we showed that acute exposure to an intermediate concentration of risperidone of 170 μg/L impaired the stress axis response, since the exposed zebrafish had lower cortisol levels than control fish, when exposed to an acute stress challenge. We also showed that zebrafish exposed to this concentration of risperidone decreased the time spent in the bottom compartment of the tank when compared with stressed fish, reinforcing the anxiolytic effect of this drug –38. To oThe risperidone mechanism is yet unclear, but it acts directly on the central nervous system, more specifically in dopaminergic and serotoninergic receptors, probably without any direct effect on interrenal tissue , 39. TheConsidering the concentrations used, the HPI impairing effect was verified only in the intermediate concentration of 170 μg/L. The lowest (0.00034 μg/L and 85 μg/L) and highest (340 μg/L and 680 μg/L) concentrations did not block the cortisol elevation after the acute stress challenge. Thus, the dose-response curve presented a U-shaped response Click here for additional data file.S2 FileStatistics of cortisol data.(PDF)Click here for additional data file.S3 FileRaw data of behavioral tests.(PDF)Click here for additional data file.S4 FileStatistics of behavioral data.(PDF)Click here for additional data file.


A 47-year-old Japanese man was diagnosed with prostate cancer in February 1995 . He underwent radical prostatectomy after androgen deprivation therapy (ADT) in June 1995. Nine years after operation, he was diagnosed with local recurrence of prostate cancer and he received postoperative external beam radiation therapy (EBRT) (70 Gy). By May 2008, the PSA dropped to 0.33 ng/mL, and a CT scan showed that the mass had disappeared. On April 2012, the PSA once again rose to 3.1 ng/mL. CT scan and MRI revealed a mass in the prostatic bed. We diagnosed local recurrence of prostate cancer. We underwent salvage low-dose brachytherapy after obtaining informed consent. The prescribed dose of the salvage brachytherapy was 145 Gy to control the tumor considering the hormone resistant prostatic cancer and high-risk feature. PSA level rapidly decreased to 0.66 ng/mL by 6 months after seed implantation. No adverse events were seen during the follow-up period. There are various methods of treatment for local recurrence after radical prostatectomy for prostate cancer. We hereby report the case of a patient who underwent low dose rate brachytherapy for castration-resistant and external beam radiotherapy-resistant local recurrence of prostate cancer after prostatectomy.A 47-year-old Japanese man was diagnosed with prostate cancer in February 1995 . He underwent radical prostatectomy after androgen deprivation therapy (ADT) at the Aomori City Hospital in June 1995. In August 2000, the prostate-specific antigen (PSA) rose to 14.7 ng/mL, and leuprorelin hormone therapy was initiated (nadir PSA: 0.5 ng/mL). In January 2004, the PSA rose to 6.1 ng/mL and therefore bicalutamide was also administered. However, the PSA rose to 12.3 ng/mL in October 2006, and a computed tomography (CT) scan revealed a mass in the prostatic bed. A biopsy from this site confirmed the recurrence of prostate cancer; thus, the patient was subjected to postoperative external beam radiation therapy (EBRT) (60 Gy). By May 2008, the PSA dropped to 0.33 ng/mL, and a CT scan showed that the mass had disappeared. Therefore, the administration of leuprorelin and bicalutamide was discontinued. On April 2012, the PSA once again rose to 3.1 ng/mL; therefore, leuplin and estramustine were administered. CT scan and MRI revealed a mass in the prostatic bed . There wPrior to salvage brachytherapy, biopsy was performed and the histological findings showed an adenocarcinoma with a Gleason score of 5 + 4. Although external beam radiotherapy of 70 Gy has been delivered prior to salvage brachytherapy, the prescribed dose of the salvage brachytherapy was 145 Gy to control the tumor considering the hormone-resistant prostatic cancer and high-risk feature. The tumor volume using transrectal ultrasound (TRUS) was 5.9 mL at a preplanning. We used a radioactive source of 0.27 mCi (11.0 MBq) according to tumor size as same as standard brachytherapy. Thirteen needles were inserted into the tumor based on a peripheral loading technique, and 2 or 3 sources were implanted through each needle. Finally 30 sources were implanted by the real-time planning method . RegardiWhen the patients show PSA recurrence or local recurrence after radical prostatectomy, there are several salvage treatments such as ADT, radiation therapy, chemotherapy, and surgical therapy –9.Our case exhibited local recurrence 11 years after undergoing radical prostatectomy and he underwent EBRT. He made a complete response; however, 6 years after EBRT, local recurrence was occurred again; therefore, low dose rate (LDR) brachytherapy was performed.MacDonald et al. investigated 102 patients who underwent EBRT for isolated PSA elevation or palpable local recurrence after radical prostatectomy [Because our patient exhibited post-EBRT local recurrence, we concerned radiation related complication after LDR brachytherapy. Lee et al. investigated 21 cases of post-EBRT salvage brachytherapy and only noted PSA failure in four patients and distant metastasis in one patient 59 months after treatment, with no adverse events of grade 3 or higher [In conclusion, we reported a case of salvage brachytherapy for castration-resistant and external beam radiotherapy-resistant local recurrence 17 years after radical prostatectomy. Longer follow-up data are needed to establish the usefulness of salvage brachytherapy for castration-resistant and external beam radiotherapy-resistant local recurrence after radical prostatectomy.


Introduction. The risk of cardiovascular disease is increased in rheumatoid arthritis (RA). A meta-analysis showed increased intima media thickness (IMT) in RA. It has been shown that disease modifying antirheumatic drugs (DMARDs) may influence the progression of atherosclerosis. However, it was suggested that biologics may be more efficient than other DMARDs (including methotrexate—MTX) in protecting against atherosclerosis. Objectives. The aim of this study was to assess the influence of different RA characteristics and treatment regimens on IMT and atherosclerotic plaques. Patients and Methods. 317 RA patients and 111 controls were included in the study. IMT was measured in carotid (CIMT) and femoral (FIMT) arteries. Arteries were screened for the presence of plaques. Results. CIMT, FIMT, and prevalence of plaques were lower in patients treated with methotrexate (MTX) ≥ 20 mg/wk, cyclosporine (CsA), or biologics than in patients treated with lower doses of MTX and other disease modifying antirheumatic drugs. No differences in IMT between patients treated with MTX ≥ 20 mg/wk, biologics, or CsA were found. Conclusions. We found a beneficial effect of MTX ≥ 20 mg/wk, biologics, and CsA on atherosclerosis. We do not confirm a stronger influence of biologics on IMT compared with therapeutic doses of MTX. Cardiovascular disease (CVD) morbidity and mortality rates are increased in RA patients compared to general population . It is eSeveral noninvasive diagnostic tools such as assessment of endothelial function, measurement of carotid intima media thickness (CIMT), and assessment of coronary artery calcification score may be used to detect subclinical atherosclerosis. A meta-analysis showed that CIMT predicts future vascular events in healthy individuals . Some stStudies in RA patients showed a decrease in flow mediated dilatation and an increase in augmentation index and pulse wave velocity, which suggests endothelial dysfunction –13. Seve2 [Several studies have focused on the influence of classical CV risk factors and disease-related factors on atherosclerosis in RA. Smoking is one of the most important CV risk factors but it is also known as a risk factor for the development of RA . Thus sm2 . A possi2 . Hyperte2 , 21. The2 . However2 , 24. Tre2 .Among RA-related factors influencing atherosclerosis inflammation seems to play a major role. Wållberg-Jonsson et al. found that high disease activity was associated with increased risk of CV event and decreased life span . A studyα therapy on subclinical atherosclerosis [α therapy reduces the likelihood of CVD in RA [α compared to those not receiving treatment; such association was not observed with other DMARDs [α compared with patients treated with MTX and other nonbiological DMARDs [It is well established that DMARDs therapy decreases CV morbidity and mortality. However, most studies focused on methotrexate (MTX) and biologics, while little is known about other DMARDs. Moreover, only few reports compared influence of different DMARDs on subclinical atherosclerosis. A prospective study by Choi et al. showed that treatment with methotrexate reduces CV mortality in RA patients . A systeclerosis –42 and sVD in RA . Interesr DMARDs . Similarl DMARDs .The aim of this study was to assess the influence of different RA characteristics and treatment regimens on CIMT, FIMT, and atherosclerotic plaques.The study was approved by the local ethical committee. All participants signed an informed consent form.317 RA patients fulfilling the 1987 ACR criteria were recruited. Exclusion criteria comprised diabetes mellitus, coronary artery disease, and history of stroke. A complete history, physical examination, and laboratory evaluation were performed and recorded in a standard protocol . All DMA111 age- and sex-matched healthy individuals were included in the control group. Clinical and laboratory data are summarized in IMT was measured on the far wall of the common carotid and superficial femoral arteries. Atherosclerotic plaque was defined as a focal structure that encroaches into the arterial lumen of at least 0.5 mm or 50% of the surrounding IMT or demonstrates a thickness of ≥1.5 mm as measured from the media-adventitia interface to the intima-arterial lumen interface. CIMT and FIMT were defined as a mean value of 6 measurements . Common carotid and superficial femoral arteries were investigated for the presence of plaques.n (%) for categorical variables. According to data distribution, a parametric (t-test) or nonparametric (U Mann-Whitney) test was used. Categorical variables were compared with chi square exact test. A P value < 0.05 was considered significant.All statistical tests were performed with STATISTICA 10.0 (StatSoft). Results are reported as mean (SD) for continuous variables and Patients and controls were age- and sex-matched. The percentage of ever-smokers, LDL, and total cholesterol concentrations and BMI were higher in controls than in RA . HoweverThe presence of plaques in RA was positively correlated with ESR, creatinine concentration, FSS, and presence of rheumatoid factor (RF) . Analysir = 0.488, P < 0.001, and r = 0.434, P < 0.001, resp.), ESR , and creatinine concentration . However, after adjustment for age, associations with creatinine became insignificant. No significant associations were found between CIMT, FIMT, and RA duration, CRP concentration, and DAS28 (data not shown).We found a positive correlation between CIMT, FIMT, and FSS (P = 1 × 10−6) and FIMT . Interestingly, RA activity (measured by DAS28) was similar in both groups: 4.64 (1.54) versus 4.83 (1.57), P = 0.3. No significant differences in classical CV risk factors were found between patients treated with MTX ≥ 20 mg/wk, biologics, or CsA. CIMT in MTX20(+)/CsA(+)/biologics(+) was comparable to controls. FIMT was slightly higher in MTX20(+)/CsA(+)/biologics(+) group than in controls. No correlations were found between CIMT and FIMT and presence of RF, ACPA, and bone erosions.CIMT and FIMT were significantly lower in cDMARDs group compared with dDMARDs group . The assRA patients are at higher risk of CVD than an age-matched general population. It is estimated that CV risk in RA is increased to a similar magnitude to that seen in type 2 diabetes . StudiesWe found a strong correlation between FSS and IMT and presence of plaques. This finding underlines the role of classical CV risk factors in pathogenesis of atherosclerosis in RA.CIMT, FIMT, and presence of plaques were associated with ESR. However, no correlation was found between atherosclerosis markers and DAS28. It may be explained by the fact that DAS28 comprises two parameters which are not completely objective: VAS and tender joints count. Moreover, swollen joints count is related to the local inflammation (synovitis), while progression of atherosclerosis in RA is thought to be due to systemic inflammation. Thus ESR, as a more objective parameter and a marker of systemic inflammation, may be a better predictor of increased risk of atherosclerosis in RA. In this context lack of association with CRP is intriguing. CRP was found to be a powerful predictor of cardiovascular disease in general population . The absCreatinine concentration was significantly higher in patients with plaques and correlated positively with CIMT and FIMT. However, after adjustment for age these associations became insignificant (data not presented). It is not surprising as a concentration of creatinine increases with age and age is a strong risk factor for atherosclerosis.α therapy on subclinical atherosclerosis [α therapy was also confirmed in meta-analyses [α and nonbiologic DMARDs; however, the authors reported a reduced risk of myocardial infarction in TNF-α-responders [α, but not in users of other RA treatments [α compared with patients treated with MTX and other nonbiological DMARDs [CIMT, FIMT, and prevalence of plaques were lower in patients treated with MTX ≥ 20 mg/wk, CsA, and biologics. This effect seems to be independent of disease activity as DAS28 was similar in MTX20(+)/CsA(+)/biologics(+) and MTX20(−)/CsA(−)/biologics(−) groups. Analysis of different combinations of DMARDs did not reveal any significant correlations (data not shown). There is a lot of evidence supporting a beneficial effect of biologics on atherosclerosis, while data concerning MTX are conflicting. Several studies showed a beneficial influence of anti-TNF-clerosis –42. A pranalyses , 50. A ssponders . Giles eeatments . Analysil DMARDs . On the l DMARDs . Our resA beneficial influence of CsA on atherosclerosis in RA is a novel finding. Few studies reported a protective effect of CsA on IMT in lupus patients . SurprisP = 1 × 10−6, and 0.081 mm, P = 5 × 10−5). A large study in general population found that an absolute carotid IMT difference of 0.1 mm is associated with a 10–15% higher risk of myocardial infarction and 13–18% higher risk of stroke [The differences in CIMT and FIMT between MTX20(+)/CsA(+)/biologics(+) and MTX20(−)/CsA(−)/biologics(−) groups were robust .


Triticum aestivum L.), Chinese Spring. In total, 416 lysine modification sites were identified on 277 proteins, which are involved in a wide variety of biological processes. Consistent with previous studies, a large proportion of the acetylated proteins are involved in metabolic process. Interestingly, according to the functional enrichment analysis, 26 acetylated proteins are involved in photosynthesis and Calvin cycle, suggesting an important role of lysine acetylation in these processes. Moreover, protein interaction network analysis reveals that diverse interactions are modulated by protein acetylation. These data represent the first report of acetylome in common wheat and serve as an important resource for exploring the physiological role of lysine acetylation in this organism and likely in all plants.Lysine acetylation of proteins, a dynamic and reversible post-translational modification, plays a critical regulatory role in both eukaryotes and prokaryotes. Several researches have been carried out on acetylproteome in plants. However, until now, there have been no data on common wheat, the major cereal crop in the world. In this study, we performed a global acetylproteome analysis of common wheat variety ( Post-translational modifications (PTMs) frequently occur to proteins during or after protein biosynthesis, and play an important role in modulating diverse cellular processes2Lysine acetylation was first discovered in histone proteins57811Advances in liquid chromatography−mass spectrometry (LC−MS/MS) and lysine-acetylated peptide immunoprecipitation enable lysine acetylation to be investigated on a proteomic scaleTriticum aestivum L.), also known as bread wheat, is one of the most important cereal crops in the world. Chinese Spring, characterized by its easy hybridization, high yielding and wide adaptability, is a specific white wheat variety in Sichuan Basin of China and has been internationally known as one of the most important wheat germplasm resources in China. The presence of a large number of acetyltranferase and deacetylase orthologs in the wheat genome suggests that lysine acetylation of proteins may play a critical role in wheat development and metabolism. In this paper, we report a proteomics study of lysine acetylation in common wheat variety, Chinese Spring. We identified 416 acetylated sites on 277 proteins controlling various biological processes in this plant. These acetylated proteins are localized in multiple compartments including nucleus, cytosol, chloroplast, mitochondria, cytoskeleton, extracellular and plasma membrane. Importantly, 26 proteins involved in photosynthesis and Calvin cycle are found to be acetylated. These results provide a systems-wide view of the wheat acetylome and an affluent dataset for functional analysis of acetylated proteins in this important cereal crop.Common wheat   . TherefoTo better understand the acetylome in wheat, GO functional classification of all the acetylated proteins was investigated based on their biological process, molecular function and cellular component . The larSubcellular localization analysis revealed that most of the identified acetylated proteins in wheat were located to the cytosol (41%) and chloroplast (36%) . FurtherIn order to further study biological processes , moleculTo better understand the general function of these acetylated proteins in wheat, they were mapped to KEGG metabolic pathways . The resFunctional enrichment analysis of identified acetylated proteins indicated that lysine acetylation may play an important role in photosynthesis. Consistent with this hypothesis, a total of 26 proteins involved in photosynthesis and Calvin cycle were found to be acetylated . In highThe cytochrome b6f complex is located in the thylakoid membrane in chloroplasts of plants, catalyzing the transfer of electrons between the two reaction complexes from Photosystem II (PS II) to Photosystem I (PS I). The results of LC-MS/MS analysis indicated that three subunits of cytochrome b6f complex , four subunits of PS II  and two ubunits of PS I (PsaA and PsaB) were among the acetylated proteins .+ oxidoreductase) catalyzes reversible electron transfer between Fd and NAD(P)HFd (Ferredoxins), one kind of iron-sulfur proteins, mediates electron transfer in a range of metabolic reactions. FNR , phosphoglycerate kinase (PGK), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), triosephosphate isomerase (TPI), fructose-1,6-bisphosphatase (FBP), Transketolase (TK) and Sedoheputulose-1,7-bisphosphatase (SBP), were found to be acetylated .Taken together, these findings support a notion that lysine acetylation potentially is an important part of the regulatory mechanism in photosynthesis and carbon metabolism.To investigate cellular processes regulated by acetylation in wheat, the protein interaction network was established . The resArabidopsis, grape, pea, potato, rice and soybean1819202122Lysine acetylation is emerging as a widespread and highly conserved post-translational modification in both eukaryotes and prokaryotes with diverse biological functions. To date, the acetylome has been reported only in a limited number of plant species including SynechocystisIn plants, one of the most important metabolic processes is photosynthesis. This process takes place in chloroplast, converting light energy to chemical energy and storing it in the bonds of sugar. Our data revealed that 100 of acetylated proteins were localized in chloroplast in wheat  and manySynechocystis sp.Another important function of photosynthesis is converting light energy to chemical energy which involves electron transfer and ATP synthesis. In this study, several components important for this function, including Cyt b6, Cyt f, Pet D, Fd, FNR and three subunit types of the ATP synthase, were all identified as acetylated proteins in wheat . In suppAs an important cereal crop in the world, wheat provides a large amount of starch ever year, which is synthesized from the raw material, starchTo summarize, our results provide the first extensive data on lysine acetylation in wheat. Our findings reinforce the notion that lysine acetylation plays a critical regulatory role in diverse aspects of cellular process, especially in photosynthesis and Calvin cycle. This study widens the range of physiological processes regulated by lysine acetylation and provides a rich resource that can be used to examine the functions of lysine acetylation in wheat and likely in all plants.T. aestivum L.), Chinese Spring, was used in the study and the seedlings were grown in a greenhouse with the temperature set at 22/18 °C (day/night) and a photoperiod of 16/8 h (light/dark)Common wheat variety (v/v) at −20 °C for 2 h. After centrifugation at 4 °C for 10 min, the precipitated protein was washed with cold acetone for three times. Finally, the protein was redissolved in buffer  and the protein concentration was estimated with 2-D Quant kit  according to the manufacturer’s instructions.Sample was grinded in liquid nitrogen. The cell powder was mixed with extraction buffer containing 8 M urea, 1% Triton-100, 65 mM dithiothreitol (DTT), 0.1% protease inhibitor cocktail, 3 μM trichostatin A and 50 mM nicotinamide4HCO3 to urea concentration less than 2 M. Finally, the diluted protein samples were digested with trypsin at 1:50 trypsin-to-protein mass ratio for the first digestion overnight, and 1:100 trypsin-to-protein mass ratio for a second 4 h-digestion.Ten mg protein was reduced with 10 mM DTT for 1 h at 37 °C and alkylated with 20 mM iodoacetamide for 45 min at room temperature in darkness. For trypsin digestion, the protein sample was diluted by adding 100 mM NH2O and the bound peptides were eluted with 0.1% trifluoroacetic acidTo enrich acetylated peptides, the tryptic peptides were dissolved in NETN buffer  followed by incubation with pre-washed agarose-conjugated anti-acetyllysine antibody beads (PTM Biolabs) at 4 °C for 12 h with gentle shakingTM Q ExactiveTM Plus). Briefly, peptides were dissolved in 0.1% formic acid (FA) and directly loaded onto a reversed-phase pre-column . Peptide separation was performed using a reversed-phase analytical column  at a constant flow rate of 300 nl/min on an EASY-nLC 1000 UPLC system. The gradient was comprised of an increase from 7% to 9% solvent buffer (0.1% FA in 98% acetonitrile) for 2 min, 9% to 24% for 24 min, 24% to 35% for 8 min and climbing to 80% in 3 min then holding at 80% for the last 3 min. The resulting peptides were subjected to NSI source followed by tandem mass spectrometry (MS/MS) in Q ExactiveTM Plus (Thermo) coupled online to the UPLC. Intact peptides were detected in the Orbitrap at a resolution of 70,000. Peptides were selected for MS/MS using NCE setting as 30; ion fragments were detected in the Orbitrap at a resolution of 17,500. A data-dependent procedure that alternated between one MS scan followed by 20 MS/MS scans was applied for the top 20 precursor ions above a threshold ion count of 1.5E4 in the MS survey scan with 30.0 s dynamic exclusion. The electrospray voltage applied was 2.0 kV. Automatic gain control was used to prevent overfilling of the ion trap; 5E4 ions were accumulated for generation of MS/MS spectra. For MS scans, the m/z scan range was 350 to 1800Three parallel analyses for each fraction were performed. The enriched peptides were analyzed using mass spectrometer  database concatenated with reverse decoy database. Peptide sequences were searched using trypsin specificity14The MS/MS data obtained above were processed using MaxQuant with integrated Andromeda search engine (v.1.4.1.2)30p value was calculated as previously describedhttp://www.ebi.ac.uk/GOA and the proteins were classified by GO annotation based on three categories: biological process, cellular component and molecular function. Kyoto Encyclopedia of Genes and Genomes (KEGG) database was used to annotate protein pathwayp-value < 0.05 were considered significant. WoLF PSORT, a subcellular localization predication program, was used to predict subcellular localization of the identified proteins. Protein-protein interactions for the identified acetylated proteins were performed using Cytoscape software and the protein-protein interaction network was obtained from the STRING database according to the methods described previously38Soft motif-x was used to analyze the model of sequences constituted with amino acids in specific positions of modifier-21-mers (10 amino acids upstream and downstream of the acetylation site) in all acetylated protein identified. Secondary structures of proteins were predicted by NetSurfPHow to cite this article: Zhang, Y. et al. Comprehensive profiling of lysine acetylproteome analysis reveals diverse functions of lysine acetylation in common wheat. Sci. Rep.6, 21069; doi: 10.1038/srep21069 (2016).


Objectives. Aluminium, a neurotoxic agent in humans, has been implicated in the pathogenesis of neurodegenerative disorders. In this study, we examined the behavioral and biochemical effects of aluminium in rats with special emphasis on memory centres, namely, hippocampus and frontal cortex. Further, the effect of simvastatin treatment on aluminium intoxication was evaluated. Methods. Rats were exposed to aluminium chloride (AlCl3) for 60 days. Simvastatin (10 mg/kg/p.o.) and rivastigmine (1 mg/kg/p.o.) were administered daily prior to AlCl3. Behavioral parameters were assessed using Morris water maze test and actophotometer followed by biochemical investigations, namely, acetylcholinesterase (AChE) activity, TNF-α level, antioxidant enzymes , lipid peroxidation, and nitrite level in hippocampus and frontal cortex. Triglycerides, total cholesterol, LDL, and HDL levels in serum were also determined. Key Findings. Simvastatin treatment improved cognitive function and locomotor activity in rats. Simvastatin reversed hyperlipidemia and significantly rectified the deleterious effect of AlCl3 on AChE activity. Further, in hippocampus and frontal cortex, aluminium-induced elevation in nitrite and TNF-α and reduction in antioxidant enzymes were inhibited by simvastatin. Conclusion. To conclude, the present study suggests that simvastatin per se protects the neurons in hippocampus and frontal cortex from AlCl3, an environmental toxin. It gets access to the human body through drinking water, food, use of utensils, deodorants, and drugs. In the brain, aluminium accumulates in sensitive areas such as hippocampus and frontal cortex and is considered a potential contributing factor to the pathogenesis of neurodegenerative disorders like Alzheimer's disease (AD) and Parkinson's disease (PD) –3. Alumiposition , 7, impaposition , apoptotposition , 10, andposition , which aposition –14.Current treatments only provide temporary and modest improvement in cognitive impairment and are considered symptomatic treatments. Thus, there is a need to develop novel and effective medication by focusing on alternative approaches, which go beyond acetylcholinesterase (AChE) inhibition and N-methyl-D-aspartate (NMDA) antagonism. Recent evidence indicates that prevalence of dementia is low among patients taking statins over a long period of time –17. ContNinety-day-old male Wistar rats, weighing 200–250 g, were used for the study. The animals were obtained from Central Animal Research Facility (CARF) of Manipal University, Manipal. For seven days prior to the experiment, the animals were acclimatized to laboratory conditions and were maintained under controlled temperature (23 ± 2°C) and humidity (50 ± 5%) conditions. Standard conditions of 12 h light/dark cycle were provided. The animals were kept in sanitized polypropylene cages having sterile paddy husk as bedding with free access to food and water. The experimental protocol was approved by the Institutional Animal Ethics Committee, Kasturba Medical College, Manipal (IAEC/KMC/73/2012), and was carried out in accordance with the guidelines given by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India.3 , simvastatin , and rivastigmine  were obtained. All chemicals used in this study were of analytical grade.Acetylthiocholine iodide, reduced glutathione, thiobarbituric acid, griess reagent, hydrogen peroxide, and 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) were obtained from Sigma-Aldrich Co. LLC . AlCln = 8). AlCl3, simvastatin, and rivastigmine solutions were made freshly each day for administration. Rats were administered with AlCl3 (175 mg/kg) orally from day 6  for 60 days. Aluminium chloride was dissolved in distilled water and administered orally at a dose of 0.5 mL/100 g bodyweight. The dose of AlCl3 was selected based on the previous literature reports [3 administration for 60 days beginning on day 6. The dose selection was based on the previous literature reports [  p.o.).Group 1: normal control—received distilled water (5 mL/kg  p.o.).Group 2: vehicle control—received 0.5% CMC (5 mL/kg 3 (175 mg/kg p.o.).Group 3: AlCl  p.o.) + AlCl3 (175 mg/kg p.o.).Group 4: rivastigmine (1 mg/kg  p.o.) + AlCl3 (175 mg/kg p.o.).Group 5: simvastatin (10 mg/kgOn probe day (day 5), the animals were randomized based on ELT (escape latency) and divided into 6 groups , southeast (SE), southwest (SW), and northwest (NW) equally spaced quadrants along the circumference of the pool. In the NW quadrant, an escape platform (10 cm diameter) was kept, 2 cm below the water surface. Throughout the acquisition trials the platform was maintained in a constant location in NW quadrant. The rats were trained to locate this hidden platform. If the rat failed to find the platform within 60 sec, it was gently guided to the platform and was allowed to stay there for 15 sec. Animals had four acquisition trials per day for four consecutive days. To eliminate the quadrant effects, animal was positioned in each quadrant during each trial. Animals that failed to reach the platform in 20 s on the 4th trial day were excluded from the study. On probe day (day 5), 24 h after the last acquisition trial, escape platform was removed and retention trial was conducted. The animals were allowed to swim for 60 s before the end of session. Retention trials were repeated on day 65 on all groups to evaluate the memory consolidation. Data were obtained through a video camera attached to a computerized tracking system  fixed above the centre of the pool. Time to reach hidden platform (escape latency), latency to find the target quadrant (northwest latency), and percentage of time spent in target quadrant (NW) were measured during retention trials.Locomotor activity was assessed in animals using a digital photoactometer . The ambulatory movements were recorded for a period of 10 min and expressed in terms of total photo beam counts for 10 min per animal . LocomotAnimals were mildly anaesthetized at the end of the experimental period with diethyl ether and the blood samples were collected by retroorbital sinus puncture into microcentrifuge tubes.Blood was allowed to clot for half an hour and then centrifuged at 10000 rpm for 10 minutes to obtain serum. Samples were divided into aliquots and stored at −20°C until biochemical analysis.The triglyceride (GPO-POD method), total cholesterol (CHOD-POD method), LDL, and HDL levels were estimated in samples by end point method as per the manufacturer's instructions with the help of diagnostic kits  using ELISA plate reader.Immediately after the probe trial on day 65, the animals were sacrificed by decapitation. On the dorsal side of the skull, an incision was made to expose and remove the brain rapidly from each rat. According to the method described by Glowinski and Iversen , hippocaThe acetylcholinesterase activity was determined quantitatively by Ellman's method . The enzThe homogenate was used for the estimation of nitrite , lipid pTotal protein was estimated in all tissue samples by using Pierce BCA Protein Assay Kit as per the experimental protocol given by Thermo Scientific, USA.α was estimated by rat TNF-α enzyme linked immunosorbant assay (ELISA) kit as per the manufacturer's instruction given by Invitrogen Corporation, USA.In the supernatant of the tissue homogenate, level of TNF-P < 0.05 were considered statistically significant.Data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. Data were expressed as mean ± standard error of the mean and the values of 3 intoxication in rats, significant spatial memory impairment was observed in Morris water maze test during the retention trial conducted on the 60th day (3 (175 mg/kg p.o.) treatment significantly (P < 0.05) raised ELT (3 was significantly (P < 0.05) reversed by simvastatin and rivastigmine  significantly (P < 0.05) produced increased locomotor activity of animals as compared with AlCl3 group.As shown in 3 produced significant (P < 0.05) upsurge in total cholesterol, triglycerides, and low-density lipoprotein (LDL) levels and significantly decreased serum high-density lipoprotein (HDL) levels compared with control group  and hippocampus (P < 0.05) of rats as compared to normal control. In hippocampus and frontal cortex, AChE activity was significantly (P < 0.05) lowered by rivastigmine compared to control group. The effect of AlCl3 on AChE activity was significantly inhibited by 10 mg/kg simvastatin  and nitrite in hippocampus and frontal cortex recorded a significant increase as compared with control group. Both rivastigmine and simvastatin (10 mg/kg) significantly reversed the elevated MDA and nitrite levels. Interestingly, simvastatin at 10 mg/kg showed a better reduction of MDA levels than rivastigmine in hippocampus region Tables  and 3.3 treated rats showed significant (P < 0.05) decrease in catalase and glutathione activity. Rivastigmine (1 mg/kg) and simvastatin (10 mg/kg) significantly improved the catalase and glutathione (GSH) levels as compared to AlCl3 treated group  and rivastigmine significantly (P < 0.05) inhibited this rise in TNF-α levels  and simvastatin (10 mg/kg) antagonized the spatial memory deficit caused by aluminium. This suggests the neuroprotective role of simvastatin in correcting cognitive dysfunction associated with aluminium exposure. Throughout the treatment period no significant changes were observed in body weight and health status of animals.Chronic aluminium exposure has been reported to result in cognitive  and loco3. These findings imply the ability of simvastatin to correct the aluminium-mediated behavioural changes.Assessment of locomotor activity is a requirement for evaluating any possible CNS depressant/stimulant effect of interventions on animals. Similar to previous reports , a decliImpaired cholinergic transmission is one of the factors implicated in the etiopathogenesis of memory deficit in AD. The neurodegeneration in frontal cortex and hippocampus areas of the brain  resultin3 inhibited acetylcholinesterase activity. Aluminium is a metal known to cause oxidative stress. Generally, oxidative stress has got the highest ability to cause damage to the highly active enzymes. Perhaps this property of AlCl3 might have produced the anticholinesterase activity observed in the present study with AlCl3. This observation corroborates with earlier workers [3) observed to have anticholinesterase activity should cause AD. We hypothesize that AlCl3 is a neurotoxin; therefore it has the propensity to cause impaired cholinergic transmission by affecting the synthesis and release. These effects are more pronounced than AChE inhibition which may be partial. This is true because the AChE inhibitory activity of standard drug rivastigmine in our study was found to be more significant than AlCl3 activity. On the other hand, simvastatin did not possess any acetylcholinesterase inhibition mechanism in vivo but abolished the effect of aluminium on acetylcholinesterase enzyme.In the present study, AlCl workers , 52 who  workers , 52, leaElevated total serum cholesterol and LDL-associated cholesterol have been implicated in AD patients –55. Even3-induced cognitive dysfunction model, elevation of reactive oxygen species (ROS) level due to significant impairment of antioxidant enzyme system has been reported [3 treatment resulted in decrease in the levels of endogenous antioxidant enzymes like catalase and GSH in hippocampus and frontal cortex regions. Further, elevation of nitrite and malondialdehyde levels was also observed in these regions of aluminium exposed rats. The alterations in the antioxidant defence system like catalase, GSH, nitrite, and MDA levels were brought to normal by both rivastigmine and simvastatin treatment. Aluminium exposure augmented the elevation in hippocampus TNF-α, a proinflammatory mediator. The elevated hippocampus TNF-α stimulates microglia to release glutamate, which causes excitotoxicity [α was counteracted by treatment with rivastigmine and simvastatin. These results indicated the potential of simvastatin in preventing oxidative stress and neuroinflammation, thereby correcting the cognitive dysfunction. Future studies will be directed to study the role of TNF-α in frontal cortex along with hippocampus to link the possible mechanism of neuroinflammation mediated cognitive dysfunction.The literature reports the contribution of oxidative stress and neuroinflammation to the pathogenesis of neurodegenerative disorders , 59. In reported . In the toxicity . The ris3-induced behavioural parameters such as cognitive deficit and locomotor impairment. Further, aluminium-mediated biochemical changes were reversed, where simvastatin was able to correct hyperlipidemia, oxidative stress, and neuroinflammation in hippocampus and cortex regions. Further studies are warranted to explore the link between AlCl3-mediated hyperlipidemia and associated dementia to establish the role of simvastatin in neuronal disturbances.Simvastatin exerted neuroprotective action against AlCl