Antibiotic pressure contributes to rising antibiotic resistance. Policy guidelines encourage rational prescribing behavior, but effectiveness in containing antibiotic use needs further assessment. This study therefore assessed the patterns of antibiotic use over a decade and analyzed the impact of different modes of guideline development and dissemination on inpatient antibiotic use.Antibiotic use was calculated monthly as defined daily doses (DDD) per 100 bed days for nine antibiotic groups and overall. This time series compared trends in antibiotic use in five adjacent time periods identified as ‘Segments,’ divided based on differing modes of guideline development and implementation: Segment 1– Baseline prior to antibiotic guidelines development; Segment 2– During preparation of guidelines and booklet dissemination; Segment 3– Dormant period with no guidelines dissemination; Segment 4– Booklet dissemination of revised guidelines; Segment 5– Booklet dissemination of revised guidelines with intranet access. Regression analysis adapted for segmented time series and adjusted for seasonality assessed changes in antibiotic use trend.p<0.001) for Segment 5 compared to Segment 4. Most antibiotic groups showed similar trends to overall use.Overall antibiotic use increased at a monthly rate of 0.95 (SE = 0.18), 0.21 (SE = 0.08) and 0.31 (SE = 0.06) for Segments 1, 2 and 3, stabilized in Segment 4  and declined in Segment 5 . Segments 1, 2 and 4 exhibited seasonal fluctuations. Pairwise segmented regression adjusted for seasonality revealed a significant drop in monthly antibiotic use of 0.401 (SE = 0.089; Use of overall and specific antibiotic groups showed varied patterns and seasonal fluctuations. Containment of rising overall antibiotic use was possible during periods of active guideline dissemination. Wider access through intranet facilitated significant decline in use. Stakeholders and policy makers are urged to develop guidelines, ensure active dissemination and enable accessibility through computer networks to contain antibiotic use and decrease antibiotic pressure. Escherichia coli resistance to ceftazidime and cefotaxime as high as 70% and 80% respectively Infections remain a major cause of morbidity and mortality in the world especially in Low-and-Middle Income Countries (LMIC) Among the factors contributing to bacterial resistance, antibiotic consumption (hereafter referred to as antibiotic use), both individual and aggregate, appear to have a major role Another purpose of antibiotic stewardship is to contain antibiotic use. Containment is important as increased antibiotic use leading to environmental pressure contributes more to bacterial resistance The present study aims to bridge this gap and determine whether antibiotic policies are truly effective in LMIC. Computerized data on medicine use has not been widely available in many of these countries. It has been however available since 2002 in the institution where the analysis was conducted. This has facilitated assessment of antibiotic use trends and patterns over the last decade through this study. The institution has taken a leading role in India for development and dissemination of antibiotic guidelines. This study has therefore seized this opportunity to assess the impact of these guidelines and its role in containing antibiotic use in hospital inpatients over this ten year period. In addition, our study aims to compare different modes of guideline implementation and dissemination and see which mode is most effective in containment. This would be important in instituting effective and sustainable antibiotic policies, especially in LMIC where there are numerous challenges. These aspects of our study are unique and the findings could be crucial to health policy makers and hospital managements.This study was carried out in a not-for-profit tertiary care teaching hospital in south India. This hospital caters to patients from various socioeconomic strata, from many parts of the country and has 2140 beds and more than 6000 outpatients per day A strict process is followed to evaluate pharmaceutical companies and their products before the medicine is stocked in the pharmacy. In addition, medicines are randomly analyzed for correct content using high performance liquid chromatography. Antibiotics in the hospital formulary are dispensed from various dispensing sections within the hospital after purchase with a prescription. Computer entry of the patient hospital number, medicine name, strength and number of units is made through the customized pharmacy software system.There were three major phases of antibiotic policy guideline implementation during the study period i.e. between July 2002 and August 2012: one in 2005 (A), the second in 2009 (B), and the third in 2011 (C). Based on this, the study period was divided into the following segments:Segment 1: July 2002 to February 2004 - This period was before the 2005 policy guidelines were introduced.Segment 2: March 2004 to December 2005 - The Antibiotic Policy Committee with active participation of clinical departments and other stakeholders such as pharmacy and microbiology department initiated preparation for a comprehensive antibiotic policy in March 2004. The preparation phase included weekly meetings and active discussion of guidelines. Guidelines were finalized and widely distributed from January 2005 as a small booklet. Active dissemination lasted till December 2005. Therefore, Segment 2 was taken as a period which included preparation and dissemination from March 2004 to December 2005.Segment 3: January 2006 to December 2008 - There was no active guideline dissemination during this period.Segment 4: January 2009 to December 2010 - Guidelines were revised and published as a booklet in January 2009 and disseminated till end of 2010. Unlike guidelines disseminated and implemented in Segment 2, there was no sustained preparatory phase and discussion with all clinical departments. This segment was therefore taken from January 2009 to December 2010.Segment 5: January 2011 to August 2012 - In January 2011, revised guidelines were published and distributed as a booklet and also made available through the intranet computer network of the hospital. Computers are available in every ward, outpatient room and departmental office of the hospital. This segment was therefore taken from January 2011 to August 2012.Antibiotic use in inpatients was calculated using the hospital pharmacy computer system. Consumption was calculated as DDD (Defined Daily Doses) normalized for 100 bed days Each antibiotic was calculated separately and coded as per DDD/ATC  Index Segmented time series design was used to assess impact of guidelines on antibiotic use across different segments. Each of the five segments in our study had a minimum of 20 monthly time points. Monthly DDD values were plotted using a standard time series plot and calendar plots to better depict monthly and seasonal variation in overall and specific antibiotic groups. The average values of DDD per hundred bed days were estimated for each segment and each antibiotic group. For exploratory purposes, we estimated the linear trend for each segment (Model 1) and examined the effect of seasonality on the segment trend for each outcome of interest (Model 2).The models were formulated as:i-segments. This exploratory analysis was performed to characterize the trend  for start and end dates for each segment.i- and i+1-segments; itime – is a month sequence (in reverse order) for i-segment; itime+1 – is a month sequence for i+1-segment; time – is a month sequence for two adjacent segments to adjust for seasonality. The difference between regression parameters i-segment, start and end of the i+1-segment, and p-values indicating the standard z-test for a linear trend against H0:i-segment and i+1-segments, respectively. For all models, the quality of fit was assessed by the R2 values. Model diagnostics showed no significant autocorrelation of residuals.In order to adjust for interruption implied by Models 1 and 2, we performed a pair-wise segmented regression adjusted for seasonality which also allows us to identify the changes in two consecutive segments Ethics permission was obtained from the Institutional Review Board of Christian Medical College Vellore, India (IRB (EC)-ER-4-10-03-2010). The analysis was done on aggregate data of antibiotic use from the pharmacy computer system of the institution. There was no direct human participation in this study and no attempt was made to identify individuals in the analysis. Informed consent was therefore waived.The time series of monthly values for overall DDD/100 bed days are shown in To facilitate visualization of trends and seasonal patterns across antibiotic groups, a compact version of monthly DDD per 100 bed days for overall and individual antibiotic groups is shown using calendar plots  and 3. Sp-values and quality of fit) and predicted values of DDD per 100 bed days for the start of the segment. The seasonality adjustment for overall antibiotic use was critical for Segments 1, 2, and 4. A significant increase of approximately 20 DDD per 100 bed days was observed in the first segment at a rate of 0.95 (SE = 0.18). Only Segment 5 showed a significant decrease in use at a rate of 0.37 (SE = 0.11) with a reduction of over 10 DDD per 100 bed days. Segment 3 however showed a significant rise in trend at a rate of 0.31 (p<0.001). As compared to Model 1, overall Model 2 improved quality of fit and better predicted DDD/100 bed days due to adjustment for seasonality (shown in bold letters).To demonstrate the trend for overall antibiotic use with and without adjusting for seasonality, the results of the two regression models are presented in The results of this exploratory analysis for individual antibiotic groups are shown in To compare the trends in adjacent segments, a pair-wise segmented regression adjusted for seasonal variation (Model 3) is presented in The study looked at ten years of antibiotic use in a tertiary care hospital, which has patients coming from all over India and beyond. During this period, antibiotic guidelines were prepared and disseminated in various modes and each had its impact in their respective segments. Overall, there was a rising trend in antibiotic use except in Segments 4 and 5 during which stabilization and decline in trend was seen . There wIndividual antibiotic groups contributed to overall antibiotic use in different ways during various periods across the decade. Before discussing the impact of guidelines on overall use, the pattern of use in each antibiotic group is therefore highlighted below  and S2.Beta-lactam antibacterials (J01C): A rising trend was observed across all segments with maximum consumption in the last segment. This could be due to the increasing use of antibiotics such as piperacillin-tazobactam for multi-drug resistant hospital acquired infections, especially in critical care settings Other Beta-lactam antibacterials (J01D): The initial segments showed a rising trend that could be explained by increasing multidrug resistance. Carbapenems are the drugs of choice for treatment of ESBL infections Quinolones (J01M): During the year, higher use was noticed in months of January and February. Quinolones are commonly prescribed empirically for respiratory infections including community acquired pneumonia and exacerbations of chronic obstructive pulmonary disease (COPD) Aminoglycosides (J01G): This group showed a rapid decline in use after Segment 3. Availability of newer antibiotics with gram negative spectrum and serious side effects of aminoglycosides like nephrotoxicity and ototoxicity Staphylococcus aureus (MRSA) may have contributed to this trend Other anti-bacterials (J01X): This group with glycopeptides and nitroimidazoles witnessed a rising trend in use. Nitroimidazoles to cover anaerobic infections and higher usage of glycopeptides to tackle rising incidence of Methicillin Resistant Sulfonamides and trimethoprim (J01E): A spike in April 2006 with rising trend in Segments 4 and 5 could be explained by additional indications like HIV associated opportunistic infections and the rise of multi-drug resistant malaria Tetracyclines (J01A): Seasonal fluctuation was observed in the calendar plot  and ModeAmphenicols (J01B): A rapid decline in use was evident. This decline was predictable after the emergence of widespread resistance especially among salmonella species Macrolides and Lincosamides (J01F): An overall rising trend with seasonal fluctuations was apparent. Macrolides, especially azithromycin, are frequently prescribed for respiratory infections which flare up during the cold months of the year nd generation cephalosporins From Swedish hospitals, an increased use of penicillins with enzyme inhibitors (J01CR) such as piperacillin-tazobactam and carbapenems has been reported. Cephalosporins on the other hand, showed a rapid decline, especially the 2Among the few studies in India, an intensive care study showed that third generation cephalosporins and meropenem were frequently used The rising trend of antibiotic use seen in Segment 1 changed as guidelines were introduced at various periods across the decade. In Segment 2, a decline in trend in antibiotic use was seen though not statistically significant  and 3. TSegment 5 was marked by dissemination of policy guidelines through a booklet and intranet access in all outpatient, departmental offices and ward computers. During this period, a significant decrease in antibiotic use was seen  and 3. IIn LMICs, the number of hospitals with hospital information system (HIS) is growing. However many do not have antibiotic policy guidelines let alone accessibility through the computer network. This strategy however should be explored by hospitals which have such capacity. Centers that can incorporate a similar mode of guideline dissemination may have a more effective strategy in containing antibiotic use as evidenced through this study. Consequently, resistance burden may reduce, ultimately leading to better healthcare and cost related benefits. A lack of infrastructure in some government facilities may hinder this strategy. Bigger government hospitals may have computer networks and therefore the situation is hopeful. Another encouraging situation is that more and more LMIC countries have fast developing information technology (IT) networks. This potential should be used by health policy makers and governments to their advantage for effective antibiotic guideline dissemination.Klebsiella pneumoniae outbreak rd and 4th generation cephalosporins and prevent increase in fluoroquinolone and carbapenem prescriptions. An intensive care unit (ICU) based prospective study done in Australia showed how implementing a computerized decision support tool resulted in 10.5% reduction of overall antibiotic consumption (166 to 149 DDDs/100 ICU bed days) The model of computer network application has been demonstrated in developed countries. A university hospital in Sweden implemented persuasive antibiotic policy guidelines in the wake of an ESBL producing The study did not assess rationality of antibiotic prescriptions and adherence of physicians to the policy guidelines. The observed trends could have been influenced by other factors due to changes in the hospital over the decade. These factors include bed capacity, number of doctors, introduction of new laboratory tests and automation, antibiotic use audits and role of other health professionals such as clinical pharmacists. However, many of these factors have played a role throughout this 10 year period and thereby their impact on decreasing antibiotic use in Segment 5 alone would be minimal. Linking antibiotic use data with resistance rates and clinical outcome such as mortality rates would have given additional information which is being planned for a future study.Overall, the rising trend in antibiotic use was contained towards the latter half of the decade. Most antibiotic groups followed this trend. Varied patterns and seasonal fluctuations were observed. The development of antibiotic policy guidelines and its dissemination contributed significantly to containing antibiotic use. The study also showed that mode of policy implementation was critical to the effectiveness of policies as formulation of appropriate policies itself. The period where guidelines were also disseminated through intranet access showed a significant reduction in antibiotic use. Where facilities are available therefore, in addition to guideline booklet dissemination by standard methods, computer networking should be used effectively to broaden the accessibility of the antibiotic policy to a much wider group of physicians and healthcare personnel. Using this type of channel will increase the flexibility for frequent updating of information, will provide additional value to physicians and a dynamic platform for effective use of antibiotics. The findings of this study would encourage hospitals in LMICs to develop and implement antibiotic policy guidelines and use modern technology for wider stakeholder access. These measures may help contain antibiotic use and thereby decrease antibiotic pressure.Table S1Exploratory analysis of trends in individual segments without (Model 1) and with (Model 2) adjustment for seasonality in nine antibiotic groups.(DOCX)Click here for additional data file.Table S2Pair-wise segmented analysis  as the estimated rate of change in monthly DDD values for adjacent segments and predicted values for the beginning of the i-segment, beginning and end of the i+1-segment.(DOCX)Click here for additional data file.


A total of 27 concrete mixtures with three replicates (81 specimens) were considered by varying the levels of key factors affecting compressive strength of concrete, namely, water/cementitious materials ratio , cementitious materials content , and fine/total aggregate ratio . The experimental data were utilized to carry out analysis of variance (ANOVA) and to develop a polynomial regression model for compressive strength in terms of the three design factors considered in this study. The developed statistical model was used to show how optimization of concrete mixtures can be carried out with different possible options.A step-by-step statistical approach is proposed to obtain optimum proportioning of concrete mixtures using the data obtained through a statistically planned experimental program. The utility of the proposed approach for optimizing the design of concrete mixture is illustrated considering a typical case in which trial mixtures were considered according to a full factorial experiment design involving three factors and their three levels (3 Optimization of the concrete mixture design is a process of search for a mixture for which the sum of the costs of the ingredients is lowest, yet satisfying the required performance of concrete, such as workability strength and durability. The basic ingredients of concrete can be classified into two groups: cement paste and aggregates. Although the quality of cement paste is governed mainly by the water/cement ratio, the quantity of cement paste required to achieve a targeted quality of concrete depends on the characteristics of aggregates. These characteristics mainly include surface area and voids in aggregates. While surface area is governed by the shape and maximum size of aggregates, the void content is affected mainly by the particle size distribution of aggregates. The requirement of the paste can be reduced by reducing the void content of aggregates through proper packing of the aggregates –5 and alAttempts have been made in the past to optimize the concrete mixture design using either the fully experimental methods or fully analytical methods or semiexperimental  methods or statistical methods. Fully experimental methods involve an extensive series of tests, sometimes conducted on a trial-and-error basis, and the optimization results are often applicable only to a narrow range of local materials , 14. In Fully analytical methods are less expensive and less time consuming but they have the disadvantage of being less precise because of the variations in the materials characteristics of the aggregates and cements. Fully experimental or semiexperimental  methods are reliable and accurate; however, they involve comprehensive laboratory works . StatistIn the present work, an effort has been made to exhibit the application of a statistical approach proposed to obtain optimum proportioning of concrete mixtures using the data obtained through an experiment design considering water-cementitious materials ratio, cementitious materials content, and fine aggregate to total aggregate ratio as design factors. The experimental data were analyzed statistically and mathematical polynomials regression was developed for concrete strength as a function of mixture variables. The utility of the developed compressive strength model in optimizing the mixture designs was illustrated considering different possible options.The proposed approach to optimizing the proportions of concrete mixtures is based on the planned experimental works (within the domain of required characteristic performance of concrete) and statistical analysis of the data generated, which would reduce the number of trial batches needed. The proposed approach consists of the following steps.3 and water/cementitious materials ratio should not be more than 0.40 (by mass) for satisfying the durability requirements for concrete subjected to severe exposure conditions such as severe freeze-thaw, deicer, and sulfate exposures [The information pertaining to required workability, strength, and exposure conditions (for durability requirements) should be first collected. The workability requirements depend on the mode of transportation, handling, and placing and also on type of construction . The strxposures .Selection of the levels of the three key mixture design factors, namely, cementitious materials content, water/cementitious materials ratio, and fine/total aggregate ratio, which mainly affect the quality of concrete will be made to ensure that enough experimental data are generated for obtaining a regression model for compressive strength which can be used to optimize the mixture proportions meeting the specified characteristic performance of concrete.3 which is the minimum value to satisfy the durability requirements for aggressive exposure conditions. The maximum level of cementitious materials content should be selected considering the risk of shrinkage. The minimum level of water/cementitious materials ratio should be selected considering the strength requirements. In case of choosing a very low level of the water/cementitious materials ratio, the difficulty in transporting, handling, and placing concrete and extra cost of superplasticizer to meet the workability requirements should be considered. The maximum level of the water/cementitious materials ratio should be within the maximum permissible limit for the water/cementitious materials ratio for the given exposure condition. The minimum and maximum levels of the fine/total aggregate ratio should be selected within the optimum range for achieving maximum packing of aggregates. For example, Soudki et al. [The minimum level of cementitious materials content should not be less than 335 kg/mi et al.  have repAn experimental work should be conducted involving designing, preparing, and testing various trial mixtures according to the full factorial experiment design considering the various possible combinations of the levels of the mixture variables within their selected ranges of variation. The workability of each trial mixture should be equal to or more than the specified value. In case if superplasticizer is needed to achieve the intended workability, the cost of superplasticizer should be added to the cost of cement. After finalizing the dosage of superplasticizer based on the required workability for each of the trial mixtures, the cubical or cylindrical specimens should be prepared, cured for 28 days, and then tested for compressive strength for generating data to obtain statistical model for strength to be used for optimization.Analysis of variance (ANOVA) can be used for examining the significance of the factors considered for developing the strength model and subsequently fitting an empirical model for compressive strength in terms of the significant mixture factors using polynomial regression. In ANOVA, the statistical terminologies used are as follows.Degree of Freedom (DF). Degree of freedom is the number of values in the final calculation of a statistic that are free to vary. DF = n − 1, where n represents the number of groups.Error . It is the amount by which an observed variate differs from the value predicted by the assumed statistical model.Sum of Squares (SS). It is the squared distance between each data point (Xi) and the sample mean (n data points. Xi represents the ith observation and Mean Square (MS). It is the sum of squares divided by the degrees of freedom. F-Ratio. It is ratio of MS of the concerned factor to the MS of the error. A higher F-ratio indicates a significant effect of the factor.P-Value. It is a measure of acceptance or rejection of a statistical significance of a factor based on a standard that no more than 5% (0.05 level) of the difference is due to chance or sampling error. In other words, if the P value for a factor is 0.05 or more, it would not have effect on the dependent variable.The statistical model for the compressive strength derived utilizing the experimental model can be used to obtain the optimal mixture proportions satisfying the specified characteristic performance of concrete as required constraints. The mixture satisfying all the constraints and having the lowest requirements of cement and superplasticizer would be considered as optimum mixture.For illustrating the utilization of the proposed approach to optimizing concrete mixture design, an experimental program was considered. A full factorial experiment with 3 × 3 × 3 treatment combinations was used resulting in a total of 27 concrete trial mixtures considering three typical levels of each of the three key factors affecting the performance of concrete mixtures, as shown in The cementitious materials used in this study consisted of 92% Type I Portland cement conforming to ASTM C 150  and 8% sThe proportioning of all 27 trial mixtures was carried out in terms of absolute volume using the specific gravities of the concrete ingredients and the values of water/cementitious materials ratio, cementitious materials content, and fine/total aggregate ratio for each of 27 mixtures, as given in Considering three replicates for each of the 27 mixtures, a total number of 81 cylindrical concrete specimens (size: 75 mm diameter and 150 mm high) were cast for determining compressive strength. After casting, the concrete specimens were cured for 28 days in a curing tank under laboratory conditions and then tested for compressive strength in accordance with ASTM C 39 [Average 28-day compressive strength test results for all 27 concrete mixtures along with the standard deviation of three replicates of each mixture are presented in Analysis of variance (ANOVA) was carried out to pinpoint the individual and interactive effects of variable factors on the dependent variable. ANOVA of the test results in the present study was done with the software named MINITAB . Based oP value was found to be less than 0.05 (95% confidence level). The P value was obtained from Fisher's distribution table which depends on error degree of freedom (DF) and the mean squares (MS). Rw/cm and RFA/TA have significant effects on compressive strength as their levels of significance, P values, are less than 0.05. Therefore, these two significant variables should be considered for obtaining the regression model for compressive strength, fc′. Although the effect of QC on compressive strength is found to be insignificant because it varies within a narrow range of 350 to 400 kg/m3, it is considered in the regression analysis as cement remains an indispensable material in concrete production.The results of ANOVA for compressive strength are presented in fc′ is the 28-day compressive strength in MPa. QC is the cementitious materials content in kg/m3. Rw/cm is the water/cementitious materials ratio by mass. RFA/TA is the fine/total aggregate ratio by mass.The polynomial regression model obtained for compressive strength using the data presented in The upper and lower bounds of each of the three variables are given in Microsoft Excel Solver:Rw/cm and RFA/TA for achieving maximum possible compressive strength at different values of cementitious materials content within the selected range ,optimizing the levels of the Rw/cm and RFA/TA for achieving different target compressive strengths at different values of cementitious materials content within the selected range .optimizing the levels of the The empirical model obtained for compressive strength can be used for optimization of concrete mixture proportions using any suitable optimization method/tool. The developed compressive strength model was utilized for optimization of concrete mixture design corresponding to the following options  typically using the 3 is higher than that at cementitious materials contents of 375 and 400 kg/m3. Further, the maximum compressive strength at cementitious materials content of 375 kg/m3 is higher than that corresponding to a cementitious materials content of 400 kg/m3. This indicates that, at the same optimum values of Rw/cm and RFA/TA, the compressive strength is more at lower cementitious materials content  due to better aggregate packing, as reported by Neville and Brooks [The optimization results, presented in d Brooks . At all 3, water/cementitious materials ratio of 0.38, and fine/total aggregate ratio of 0.45 can be typically selected as the optimum mixture. However, in cases where the compressive strength requirement is less than the maximum, a set of water/cementitious materials ratio, cementitious materials content, and fine/total aggregate ratio other than the optimum one can be selected for achieving the workability and durability requirements.From The data obtained from the optimization option (ii), as presented in A simplified step-by-step approach is proposed for optimizing the concrete mixture design based on the analysis of the data obtained through a statistically planned experimental program. The proposed approach consists of five steps, as follows: (i) specification of the characteristic performance of concrete, (ii) selection of the levels of key mix design factors, (iii) experimental work considering trial mixtures using full factorial experiment design for generating data to obtain statistical model for optimization, (iv) statistical analysis of experimental data and fitting of the strength model, and (v) optimization of mixture proportions using the fitted strength model.The results of the experimental works conducted in the present study for demonstrating the utility of the proposed statistical approach have indicated the significant effects of water/cementitious materials ratio, cementitious materials content, and fine/total aggregate ratio on compressive strength. The optimum values of water/cementitious materials ratio and fine/total aggregate ratios have resulted in a higher compressive strength at a lower cementitious materials content resulting in significant cost saving in the concrete production.


Filarial parasites are known to induce a large range of immunoregulatory mechanisms, including the induction of alternatively activated macrophages and regulatory T cells. These mechanisms are used to evade and down‐modulate the host’s immune system, to support parasite survival. Several reports have focused on some of these mechanisms, in humans and murine models, but the complex immunoregulatory networks associated with filarial infections remain unclear. Recent publications have conferred a role for regulatory T cells in the ability of helminth parasites to modulate human immune responses, such cells promoting the induction of the non‐complement‐fixing immunoglobulin G4 (IgG4). High plasma concentrations of IgG4 have been reported in hypo‐responsive and asymptomatic cases of helminth infection. In both human lymphatic filariasis and onchocerciasis, the asymptomatic infections are characterised by high plasma concentrations of IgG4 (compared with those of IgE) and of the complement‐fixing antibodies IgG1, IgG2 and IgG3. In asymptomatic filarial infection, elevations in IgG4 are also often associated with high worm loads and with high plasma levels of the immunomodulatory interleukin‐10. Here, various aspects of the induction of IgG4 in humans and it roles in the immunomodulation of the human responses to filarial parasites are reviewed. Wuchereria bancrofti, Brugia malayi and B. timori), onchocerciasis (Onchocerca volvulus) or loiasis (Loa loa). These infections are often chronic and persist for many years. In most infected individuals, however, few, if any, symptoms develop, the relatively asymptomatic existence of filarial parasites in the host being the result of the ability of filarial nematodes to down‐modulate the host’s immune responses, in order to facilitate their own survival. Unfortunately, some infected individuals do develop serious immunopathology. The immunological and genetical factors influencing the direction that any filarial infection takes, towards immunopathology or to tolerance, are still not fully understood.Infections with filarial nematodes remain a major public‐health problem, especially in tropical countries , >200 miInduction of the non‐cytolytic and blocking immunoglobulin G4 (IgG4) is believed to represent one major mechanism used by filarial parasites to evade destruction by their host’s immune system. The immunoregulatory properties of IgG4 have been recently demonstrated , the funin vitro and in vivo  were generally amicrofilaraemic and had substantially higher levels of IgG1, IgG2 and IgG3 but, on average, 3.4‐fold lower levels of specific IgG4 than the asymptomatic microfilaraemics. W. bancrofti filariasis, Human lymphatic filariasis, also known as elephantiasis, is caused by three species of filarial parasites: O. volvulus. The adult parasites reside mainly in subcutaneous nodules  illustrated the prominence of IgG4 in human onchocerciasis and the role of such immunoglobulin in the survival of microfilariae. Dafa’alla et al. (1992) compared the IgG responses to O. gutturosa and O. volvulus and concluded that IgG4 secretion, in response to O. volvulus or O. gutturosa, was considerably higher than IgG1, IgG2 or IgG3 secretion . They also demonstrated that IgG4 levels were positively correlated with microfilaraemia, whereas IgG3 levels showed a negative association with the microfilarial load. The high levels of IgG4 antibody in individuals with high microfilarial loads indicate a role for this isotype in inhibiting microfilarial clearance. Dafa’alla et al. (1992) suggested that IgG4 competed with other antibodies, probably IgE and IgG3, for adherence to the microfilariae and thereby inhibited the ADCC known to be involved in microfilarial destruction , L. loa were significantly higher in the amicrofilaraemic subjects than in the microfilaraemic. These observations indicate that L. loa microfilariae are at least partially responsible for the preferential production of IgG4 in human loiasis. The absence of microfilariae is often associated with the production of the more immunocompetent immunoglobulins IgG1 and IgE, which often appear associated with the development of immunopathology. Curiously, in an earlier study in Gabon by the same research group, similarly high levels of IgG4 expression were found in subjects with and without L. loa microfilaraemias  as well as IL‐10‐producing and CD4(+)CD25(+)FOXP3(+) regulatory T cells , microfiIn their recent study, It may be the IgG4/IgE or IgG4/IgG ratio that is one of the main criteria determining whether a filarial infection heads toward immunoregulation (and an asymptomatic state) or immunopathology. Hence, IL‐10 from microfilariae‐induced regulatory T cells might, in the context of an immunological synapse, induce B cells to secrete IgG4 preferentially, promoting parasite tolerance. Toll‐like receptors that are known to be expressed on regulatory T cells CD25(+)FOXP3(+) regulatory T cells and antigen‐induced, IL‐10‐producing, type‐1 regulatory T cells modulate the host’s immune responses by enhancing the production of non‐cytolytic IgG4 antibodies. The IgG4 molecules are capable of inhibiting IgE‐ and IgG‐mediated effector mechanisms. This humoral regulation makes an important contribution to the avoidance of pathology . In the absence of immunoregulation, the host’s immunocompetent antigen‐presenting cells activate effector T‐cells, which, in turn, induce B cells to produce cytolytic antibodies . These antibodies, through different effector mechanisms (e.g. complement activation and ADCC), induce parasite death and the subsequent release of antigens from endosymbobiotic logy see . A bette


This in vitro study was designed to compare enamel demineralization depths adjacent to bands cemented with zinc polycarboxylate, glass ionomer (GI) and resin-modified glass ionomer (RMGI), in order to achieve minimal enamel demineralization during orthodontic treatment.Sixty fully developed extracted third molars were randomly divided into three testgroups each containing 20 samples, used to cement orthodontic bands with zinc polycarboxylate, GI and RMGI. All samples were demineralized using White method using hydroxyapatite, latic acid and Carbapol for in vitro caries simulation, and then, immersed in 10% solution of methylene blue. The mean depth of dye penetration was assessed up to 0.1 millimeter, reflect-ing the depth of enamel demineralization. One way ANOVA and LSD statistical tests were employed to evaluate significant differences among groups.The highest dye penetration depth was seen in zinc polycarboxylate group, followed by GI, and RMGI groups, respectively, with significant differences among each two groups (P < 0.05). The use of RMGI cement seems to present significantly better prevention of enamel demineralization adja-cent to orthodontics bands. The bacteria metabolize carbohydrates and produce organic acids, dissolving the calcium phosphate mineral of tooth structures.4 Different studies have reported the prevalence of these lesions up to 95% during fixed orthodontic treatment.7 Demineralization can occur within few weeks,8 a length of time that is usually shorter than preferred by most orthodontists.10 It has been generally accepted that, the combined application of fluoride regimens, oral hygiene instructions and dietary control can contribute greatly to inhibition of demineralization during fixed appliance treatment. However, these preventive strategies need patient compliance,16 and hence, are not reliable.19Enamel demineralization adjacent to bands and brackets is a great complication in the patients of fixed orthodontic treatment especially those with poor oral hygiene.21 The professional application of fluoride varnish has been shown to be effective in this situation.22 Fluoride varnish adheres to enamel surface longer than other topical fluoride products and has been shown to be superior because of its ability to increase fluoride uptake in enamel in vitro.23 Also, fluoride has been added to the materials used for cementation of bands and brackets. The cement acts as an additional local source of fluoride near the appliances, whereas the amount of release depends on the substance group each material belongs to.24Therefore, preventive strategies which do not need the patient’s compliance might be more effective in preventing or reducing demineralization.2 Resin- modified glass ionomer cements have higher adhesive strength than conventional glass ionomer and can also control caries around orthodontic appliances.27 Although previous studies have evaluated the efficacy of different cements, few have compared the efficacy of different routinely-used materials for cementation of orthodontic appliances.28 Therefore, the present study was designed to compare zinc polycarboxylate, conventional glass ionomer, and resin-modified glass ionomer cements with regard to enamel demineralization adjacent to orthodontic bands after in vitro pH cycling.Conventional glass ionomer cement has been evaluated for bonding orthodontic brackets due to its anti-cariogenic property attributed to release of fluoride.Fully developed extracted third molars with same morphology and size were included in this study. The teeth were collected from Department of Oral & Maxillofacial Surgery, Faculty of Dentistry, Shahed University, and two private dental clinics in Tehran, Iran. Written consents were obtained from all patients. The teeth were assessed precisely and those with enamel hypoplasia, developmental malformations, discolorations and any clinical evidence of dental caries were excluded from study. Then, the sixty selected teeth were cleaned with fluoride-free pumice by low-speed handpiece to remove any debris, rinsed with deionized water, and randomly divided into three test groups, each containing 20 samples. All teeth were embedded in acrylic blocks.18 In this manner, the enamel both directly in contact with cement and up to 2 mm away from the cement had similar situation for induction of in vitro demineralization. Therefore, it would be an internal control for testing of in vitro pH cycling performance.Stainless steel microethced orthodontic bands  with attachments were fitted on teeth, and margins were adapted by band pusher. The bands were approximately seated at the same position of each tooth on middle third part of crowns. Then, bands were tightly fitted to decrease the possibility of enamel dissolution. After manipulating, the bands were cemented on teeth in each group using one the following materials according to the manufacturers’ instructions: zinc polycarboxylate cement ; glass ionomer (GI) cement ; resin-modified glass ionomer (RMGI) cement . RMGI was polymerized with curing light, from two opposite sides of the bands for 40 seconds, to ensure adequate polymerization. In all samples, additional cement was removed before polymerization from occlusal and cervical margins of the bands to prevent affecting test results. The cements were allowed to bench set for 2 minutes at a uniform ambient temperature of 25ºC. Then, to prevent demineralization of tooth body, each tooth was coated with two layers of nail varnish, except a 2×2 mm window under cervical margin of the band, which was designated for evaluation of demineralization.12 as follows: the samples were demineralized in 10 mL of an acidic solution (pH = 4.8) at 37°C, containing 500 mg per liter of hydroxyapatite, 0.1 mol per liter of lactic acid and 20 g per liter of carbopol 907; the solution was changed every 4 hours for 96 hours.12 Then, they were removed and rinsed with deionized water. The use of colored agents to demonstrate microleakage continues to be the most popular of techniques currently available. This method allows the production of sections showing leakage in contrasting colors to the tooth and without the need for further chemical reaction or exposure to potentially hazardous radiation.29 So, in the next step, teeth were immersed in a 10% solution of methylene blue at 37°C to evaluate the created demineralization through dye penetration.11 By then, the teeth were removed and rinsed with deionized water. After removing bands by band remover, samples were sectioned buccolingually through the midline of exposed window by disk (Isomet low speed saw and wafering blades).Imbibition of a dye into porosities of demineralized enamel enables quantitative analysis of demineralization by stereomicroscope (Ruler-JY 80X) with ×80 magnification. The depDescriptive statistics, including means and standard deviations, were calculated for dye penetration in all study groups. Then, results were analyzed by one-way ANOVA to detect significant differences among all groups. Then, multiple comparison LSD tests were applied to identify significant differences for dye penetration between each two groups. The statistical significance level was set at P < 0.05.The mean dye penetration of all groups is given in30 Therefore, using fluoride releasing cements for band cementation would be effective in preventing enamel decalcification.Clinical experience shows that the orthodontic band makes tooth more prone to decalcification. This is because the orthodontic band with its welded attachment provides a place for plaque accumulation and when the band is not fully fitted onto the tooth, there will be a gap between the band and the tooth which makes the tooth more prone to decalcification. Studies have shown 85% of the cervical and occlusal margins of the band present cement defects.31 The lowest demineralization observed around orthodontic bands cemented with RMGI is consistent with the results previous studies.,3 Therefore, considering the use of RMGI as a preventive orthodontic banding cement might be recommended pending further investigations.In this study, there were significant differences among demineralization depths of zinc polycarboxylate, GI and RMGI groups. The differences might be justified by the higher amount of fluoride released by RMGI as well as its appropriate bond strength and lower solubility.34 with findings similar to those of the present study. High demineralization around this cement might be related to its physical properties, as research on zinc polycarboxylate cement has shown poor bond strength to enamel and dentin as well as high solubility.3 Although the experimental protocol was similar to those in previous studies, an attempt was made to provide a similar situation for all groups in the present study to reduce the effect of these factors. Indeed, the time taken for causing demineralization was as long as 3 months of real time,12 compare to most studies. However, with regards to studies showing the high amount of fluoride releasing in first three days from cementing and then a substantial decrease after the third week,19 evaluating the effect of these cements for longer period would not be efficient. Therefore, we tried to improve the reliability of the study by increasing the volume of the samples, compared with previous studies.The highest demineralization adjacent to bands was seen in zinc polycarboxylate group (P<0.05). Evaluation of demineralization adjacent to zinc polycarboxylate cement has been done before,Although in vitro studies can be helpful in evaluating the demineralization properties of polycarboxylate, glass ionomer and resin-modified glass ionomer cements, laboratory conditions cannot replicate the exact clinical condition. Further controlled clinical trial is warranted.With regard to limitation of an in vitro setting, it can be concluded that the use of RMGI would provide significantly better prevention of enamel demineralization adjacent to orthodontic bands.


We discovered that rbf1 mutations enhance developmental phenotypes caused by Psc overexpression such as ectopic cell death and disorganized ommatidia. Our genetic analysis revealed that Psc-induced developmental defects are strongly influenced by CAF1p55, which is a shared component of several chromatin-associated complexes including a histone chaperone complex, chromatin assembly factor-1 (dCAF-1). Interestingly, the expression levels of dCAF-1 components, CAF1p105 and CAF1p180, are increased in rbf1 mutants, whereas the expression level of CAF1p55 itself remains relatively unchanged. We demonstrated that the increased levels of CAF1p105 and CAF1p180 are required for the hypersensitivity of rbf1 mutant cells to Psc-induced cell death and for the developmentally regulated cell death normally observed in rbf1 mutant eyes. We propose that Caf1p105 and Caf1p180 are important determinants of cell death sensitivity in rbf1 mutant cells and contribute to the genetic interaction between Psc and rbf1.A Polycomb group protein, Posterior sex combs (Psc), was identified in a genetic screen designed to find factors that can specifically induce morphological defects in  Retinoblastoma protein (pRB) is a tumor suppressor protein that was first discovered as a result of its deletion in rare childhood retinal cancers called retinoblastomas . The rolp53 gene are not found, a negative regulator of p53, MdmX, is amplified to inactivate the p53 pathway (Syk) is increased by epigenetic changes in retinoblastomas and is required for the survival of retinoblastoma cells, making Syk a potential therapeutic target , which first dimethylates and trimethylates histone H3 lysine 27 (H3K27). The trimethylation of H3K27 is then recognized by PRC1, which is thought to maintain epigenetic silencing by compacting chromatin and inhibiting ATP-dependent nucleosome remodeling. Among the PRC1 components, Psc is shown to inhibit transcription and chromatin remodeling in vitro, independently of the other PRC1 components (Polycomb group (PcG) proteins are epigenetic repressors that maintain gene silencing of important regulators of development, such as Hox genes. PcG proteins form evolutionary-conserved multimeric polycomb repressive complex 1 and polycomb repressive complex 2 (PRC1 and PRC2) that can function together to silence PcG target genes . In Drosmponents .A PRC2 component, CAF1p55, is also part of several other chromatin-associated protein complexes, such as nucleosome remodeling factor, NuRD, dREAM/MMB, and a histone chaperone complex, chromatin assembly factor-1 (dCAF-1). CAF1p55 seems to function as a noncatalytic component of these complexes by promoting chromatin association through binding to histones H3 and H4  from a genetic screen designed to identify factors that can cooperate with rbf1 mutations  is a factor that, when overexpressed, can interfere with cell survival and differentiation in rbf1-deficient eyes. Further analysis revealed that rbf1 mutations provide a sensitized genetic background for Psc-induced developmental defects. Our genetic studies revealed that CAF1p55 function is commonly affected by Psc overexpression and rbf1 mutations. Interestingly, the expression of dCAF-1 components, CAF1p105 and CAF1p180, is upregulated in rbf1 mutants, whereas the expression level of CAF1p55 remains relatively unchanged. Importantly, we demonstrate that Caf1p105 and Caf1p180 are required for the developmentally regulated cell death phenotype in rbf1 mutant eyes and the Psc-induced cell death phenotype.In the same genetic screen that identified 120arbf1 is a hypomorphic rbf1 allele that was previously described , which removes an FRT cassette to allow Gal4 expression from the Caf1p55 (stock 105838), Caf1p105 (stock 110461), and Caf1p180 (stock 108240) RNAi alleles were obtained from the Vienna Drosophila RNAi Center. The following mutant alleles were obtained from the Bloomington Stock Centre: 731 FRT2a/TM6cE(z); FRT1/TM6C82B Sce; KG0335ISWI; and EY0530/CyO. escKG07458 40A/CyOmip120 was provided by the Drosophila Genetic Resource Center at the Kyoto Institute of Technology. long, FRT82B Caf1med, FRT82B Caf1short,FRT82B Caf1 and UAS-Caf1p55 flies were provided by Dr. Graeme Mardon (Baylor College of Medicine).longCaf1p55 cells were generated by mitotic recombination using ey-FLP and FRT 82B P(W+) l(3)cl-R3, which carries a recessive lethal gene. For rbf1 mutant clones, Δ14rbf1 was used to create rbf1-null clones.Eye imaginal discs composed largely of homozygous Clones in the following genotypes were analyzed:longyw ey-FLP/Y;FRT 82B P(W+) l(3)cl-R3/FRT82B Caf1p55120aey-FLP/Y;FRT 82B P(W+) l(3)cl-R3/FRT82B Caf1p55longrbf1Δ14rbf1, ubi FRT19AFRT19A/GFP; ey-FLPFor immunostaining, third instar larval and pupal eye imaginal discs were fixed and stained as previously described . RNA was treated with on-column RNAse-free DNase I digestion (Qiagen) to ensure the elimination of genomic contamination. cDNA was synthesized from 500 ng RNA using the DyNAmo cDNA Synthesis Kit (Finnzymes) with random hexamers. Gene expression was measured in triplicate using DyNAmo Flash SYBR Green qPCR Kit (Finnzymes) and normalized with reference genes plicates . Primersplicates  and are rp49 forward TACAGGCCCAAGATCGTGAAGrp49 reverse GACGCACTCTGTTGTCGATACCβ-tubulin forward ACATCCCGCCCCGTGGTCβ-tubulin reverse AGAAAGCCTTGCGCCTGAACATAGCaf1p105 forward ACTCGCTTGGTATTGGCATCCaf1p105 reverse CAGTGGCAAATCACTGGCTACaf1p180 forward GTCGGCGAAATGCAGATACTCaf1p180 reverse AAGGACGAGGAGGATGATGACaf1p55 forward CATCAAAAGGAAGGCTACGGCaf1p55 reverse GCCGGTGAAGATGTTCTTGPsc forward CATCAGTTCCCGTTCGTAAAGPsc reverse CGAGATGGTCATCAACAACG120arbf1 hypomorphic flies . f00391 induced small, disorganized, and rough adult eyes in rbf1 hypomorphic flies but failed to produce such defects in wild-type flies  showed that the characteristic stripe of cell death in 120arbf1 eye discs at the morphogenetic furrow was enhanced by f00391 but resulted in no substantial change in a wild-type background . This observation led us to hypothesize that 4 driver . Even iny f00391 . ELAV anll death . NeverthCaf1p55 mutant alleles. Interestingly, the loss-of-function phenotypes observed in Caf1p55 mutant eyes are similar to those induced by Psc overexpression. Moreover, reduced Caf1p55 activity leads to arista-to-tarsi transformation, a documented phenotype of Psc overexpression that we have observed with f00391 . To further investigate the relationship between Psc and Caf1p55, we took advantage of the fact that a stronger driver, GMR-Gal4, with f00391  can produce a dominantly modifiable rough eye phenotype in a wild-type background. Although GMR-Gal4;f00391 failed to recapitulate the defect observed with ELAV staining, the same Senseless expression defect could be observed when Psc was overexpressed with GMR-Gal4 . Importantly, the adult eye phenotype of GMR-Gal4;f00391 was suppressed by a single copy of an Sce mutant chromosome, which, like Psc, is a component of PRC1 . This result demonstrated that the rough eye phenotype of GMR-Gal4;f00391 could be used to identify factors that can dominantly modify the Psc-induced phenotype. Anderson et al. identified three Caf1p55 alleles, shortCaf1p55, medCaf1p55, and longCaf1p55. The shortCaf1p55 and medCaf1p55 alleles have point mutations that introduce stop codons in the CAF1p55 coding region, whereas the longCaf1p55 allele carries a point mutation that replaces the Gly375 with an Asp . Although the molecular nature of the mutation is unclear, one possibility for this observation is that the gene product of longCaf1p55 retains some CAF1p55 function that is able to resist Psc overexpression. We then determined the effect of simply coexpressing wild-type CAF1p55 on the GMR-Gal4;f00391 rough eye phenotype. As shown in GMR-Gal4;f00391–induced rough eye phenotype. Coexpression of the baculoviral caspase inhibitor p35 was unable to suppress the eye phenotype, indicating that the ability of CAF1p55 to suppress the rough eye phenotype is specific . Because it remains unclear how the point mutation in longCaf1p55 allele affects overall CAF1p55 function, we were unable to draw strong conclusions from the lack of discernible phenotypes. This result raises the possibility that the normal function of Caf1p55 is compromised by rbf1 mutations, providing a plausible explanation for the sensitivity of rbf1 mutant cells to Psc overexpression.The genetic interaction between ant eyes . Third i mutants . InteresCaf1p55 and rbf1 directed us to explore ways that CAF1p55 function might be affected by rbf1 mutations. Aside from its function as a PRC2 component, CAF1p55 is part of many chromatin-associated complexes, including nucleosome remodeling factor, RBF-containing dREAM/MMB, and dCAF-1 , which was able to substantially decrease CAF1p180 protein expression . Strikingly, coexpression of iCaf1p180 strongly suppressed the cell death phenotype caused by f00391 in 120arbf1 eyes, indicating that Caf1p180 is required for sensitivity of rbf1 mutant cells to Psc-induced cell death , indicating that Psc overexpression interferes with broad developmental processes during Drosophila eye development. Given the scale of the biological processes in which PcG proteins participate, these findings were not unexpected . However, without delving into the exact molecular mechanism, our genetic study does not address whether all CAF1p55-associated complexes are equally affected by Psc overexpression. Moreover, the Senseless expression defect caused by Psc overexpression was not observed in the Caf1p55 mutant study by Anderson et al. indicating that Psc overexpression may have differential effects on CAF1p55-associated complexes or CAF1p55-independent effects. Interestingly, the longCaf1p55 allele was able to dominantly suppress the GMR-Gal4;f00391 rough eye phenotype, whereas a stronger Caf1p55 allele, shortCaf1p55, could not . Importantly, Caf1p55 overexpression did suppress the Psc-induced cell death phenotype in an rbf1 mutant background, again supporting the notion that Psc and Caf1p55 can antagonize each other when they are overexpressed . An alternative model to explain the effect of Caf1p180 and Caf1p105 depletion on the rbf1 mutant cell death stripe in eye discs is that deregulated dCAF-1 activity may function independently of other CAF1p55-associated complexes. dCAF-1 has been shown to be involved in transcriptional regulation such as in heterochromatin formation . However, because Caf1p55 depletion itself induces a strong cell death phenotype in both control and rbf1 mutant eyes, we could not make a strong conclusion from this result. Whether deregulated dCAF-1 activity can affect the function of other CAF1p55-associated complexes, our genetic data clearly show that increased expression of CAF1p180 and CAF1p105 is necessary for the ability of Psc to promote cell death in rbf1 mutant cells. Interestingly, overexpression of CAF1p180 has been shown to induce cell death during Drosophila eye development (Psc and rbf1 because the differentiation defects were not suppressed by Caf1p180 or Caf1p105 RNAi (We demonstrated that the increased expression level of CAF1p180 and CAF1p105 contributes to the genetic interaction between nt cells . One int105 RNAi .Previous studies have focused mainly on the direct targets of E2F transcription factors to understand the transcriptional consequence of pRB deficiency. Our study provides evidence to suggest that pRB deficiency deregulates CAF1p55 and possibly alters the activity of CAF1p55-associated protein complexes. Interestingly, a recent study showed that retinoblastoma genomes are more stable than previously thought, whereas a large number of epigenetic changes are commonly found in these tumors (


C. bombicola (ATCC22214). This is useful towards lowering the SL production cost. Through optimization of fermentation parameters and use of resting cell method, the yield 15.25 g/L could be achieved for Jatropha oil derived SL (SLJO) with 1% v/v oil feeding. The synthesized SL displayed good surfactant property. It reduced the surface tension of distilled water from 70.7 mN/m to 33.5 mN/m with the Critical Micelle Concentration (CMC) value of 9.5 mg/L. Keeping the prospective use of the SL in mind, the physicochemical properties were checked along with emulsion stability under temperature, pH stress, and in hard water. Also antibacterial action and stain removal capability in comparison with commercial detergent was demonstrated. SLJO enhanced the detergent performance. Based on the results, it can be said that SLs have utility as fabric cleaner with advantageous properties such as skin friendly nature, antibacterial action, and biodegradability. Therefore SLs are potential green molecules to replace synthetic surfactants in detergents so as to reduce harm caused to environment through detergent usage.Sophorolipids (SLs) are glycolipidic biosurfactants suitable for various biological and physicochemical applications. The nonedible Jatropha oil has been checked as the alternative raw material for SL synthesis using  In terms of production volume, surfactants belong to the most important classes of industrial chemicals with a current total world production exceeding 13 million tonnes per year . The ecoC. bombicola, Candida apicola, and Candida bogoriensis  of the expected forms of SLs. Different forms of SLs derived from palmitic , stearic , oleic , linoleic , and trace amounts of arachidic  acid were detected. In the synthesized SL, diacetylated SL of C18:1, that is, oleic acid m/z 711, was detected as the most abundant structural form. Constituent structural forms corresponding to the observed m/z values have been listed in Typical structure of SLs consists of a sophorose (dimeric sugar) linked hs 16–18 . SL occu1H NMR spectrum of the SLJO preparation was assigned to a typical glycolipid-type structure. And characteristic proton chemical shift peaks could be observed. Protons of (–CH3) of fatty acid resonated at 1.20. Protons of (–CH2) bonded to carboxylic group of fatty acid resonated at 1.99. Resonance of protons belonging to sophorose moiety resulted in peaks within the region 4–4.5. Appearance of peaks around 5.3 was attributed to the signals from (–CH=CH–), that is, unsaturation in the fatty acid chain. The data was found to be in agreement with previously reported SL-NMR data from relevant references [The ferences , 29.Before proceeding to further analysis, surfactant property of the synthesized SL was qualitatively confirmed by oil displacement test. The surface active properties were checked using different experimental and analytical methods.It was seen that SLJO reduced the surface tension of distilled water from 70.714 mN/m to 33.512 mN/m at the CMC value of 9.5 mg/L as depicted in The emulsification activity and stability of SLJO and synthetic surfactants, namely, SDS and Triton X-100, have been noted in As an ingredient of detergent, SLJO has to perform satisfactorily in extreme physical conditions and different water qualities. Therefore the effect of different parameters on emulsifying property was explored.(a) Effect of Water Hardness on Emulsifying Property. Results mentioned in (b) Effect of pH on Emulsifying Property. pH is known to be one of the most important environmental factors influencing the performance of any biosurfactant. pH alters the net charge on surfactant molecule and thus its orientation at the interface.A600 values. It was observed that emulsification activity of SLJO was comparable between pH values of 5.0–8.0 while the emulsification index value at pH 4.0 was 6 times low as compared to other pH values. Classically detergent formulations make the pH alkaline. SLJO was found to retain the surfactant activity at alkaline pH which makes SL suitable for combination with commercial detergents.The experiment was repeated twice and the emulsification indices were calculated from the average It was observed that emulsions formed by SLJO were stable within the pH range of 5.0–8.0. The decay constant of SLJO is −11.3184 at pH 4.0 (refer to supplementary information for details). The results are in agreement with those of the report by Daverey and Pakshirajan, 2009, wherein they have used sugarcane molasses and soybean oil for SL synthesis and found that emulsifying activity was maximum at pH 7.0 and highest stability at pH 8.0 .(c) Effect of Temperature on Emulsifying Property. It was observed from SLs are known to possess antimicrobial properties . The pro90 values were determined. Against Staphylococcus aureus, MIC90 of SLJO was 300 μg/mL, while against Escherichia coli SLJO could not achieve 90% inhibition till 500 μg/mL. Thus it can be said that SLJO is more active against gram positive index bacterium. The antibacterial character of SLJO is an additional advantage while using it in detergent formulations  and stainless steel (85° to 42°). No much change in contact angle was seen in case of glass surface. Thus water droplets will spread evenly on teflon and stainless steel to give low θ between the solid and a drop of a liquid applied to its surface is formed and this can be taken as a measure of wetting [Dose dependent wetting performance of SL was assessed using canvas disc method. At 0.01% concentration of SL, 8.8 minutes were required for sinking, while at 1 g% concentration sinking time decreased to 1.15 minutes. In the first phase of washing, textile fibres and soil must be wetted as thoroughly as possible by the wash liquor. Wetting is a complex process which is determined by the interaction of the different interfacial tensions between the solid surface, the liquid, and the gas phase. A contact angle  wetting . From thSinking time required in case of combinations of SLJO with SDS and Triton X-100 was less than the time required for individual SLJO, SDS, and Triton X-100 refer to . With inSimilar results were observed in case of remaining 3 types of stains, turmeric, oil, and poster color . In cases where total cleaning was not achieved with the prescribed washing protocol, visibly SLJO performance almost matched that of the commercial detergent. And the combination SLJO-detergent worked best.In the detergency test, stains differing in their chemical nature have been used which are considered to be notorious such as caffeic acid, a yellow solid containing phenolic, acrylic group in coffee stain, and curcuminoids in turmeric. Conventionally bleach or acids are used for these kinds of tough stains which damage the fabric; on the other hand, SLs are skin friendly.Hence the results can be summed up as there is an indication that, for majority of stains, SLJO can work as good as detergent. And through standardization of the washing procedures, there can be improvement. When combined with detergent, SLJO enhances their action. This way, reduction in the detergent load to half is really attractive and will have big positive impact.Jatropha curcas. The present report can be regarded as one of the first reports with reference to utilization of nonedible oil for SL production. The yield value observed was 15.25 g/L with optimized conditions and 1% v/v oil feeding. The Jatropha oil contains alkaloids and phenolics. Thus detoxification has been achieved by exploiting the robust organism, C. bombicola, and a valuable molecule has been produced from renewable stock. Resting cell method has been used in contrast to growth associated SL production which improved production economics.In the present paper SL, a type of biosurfactants, has been produced using nonedible Jatropha oil derived from seeds of The SLJO was found to work at low CMC value, that is, 9.5 mg/L. Other desirable properties of surfactants to work as good detergent such as wetting property, contact angle reduction, antibacterial action, and so forth have also been confirmed with the SLJO. Emulsification property of SLJO has been evaluated with special reference to changes in environmental parameters pertaining to different water qualities. Also we are reporting the use of SL for cleaning the fabric stains in comparison with commercial detergent formulation. SL acts as efficient stain cleaner. When used with detergent, it showed improved performance, thus reducing the load and harm caused to environment. This is specifically advantageous as the half-life of detergent can be up to 16 days which is detrimental to aquatic life and badly affects the ecological balance of water bodies. On the contrary, the biosurfactant SLs are biodegradable, ecofriendly, and nontoxic.The supplementary information contains detailed data on SL yield maximization i.e. response to variation in media composition and fermentation parameters as described in section 2.3, NMR pattern of SLJO, detailed surface tension reduction behaviour of various concentrations of SLJO. These are followed by details about emulsifying activity assessment, contact angle reduction and results of wetting property assessment of individual surfactants. The results of detergency test against remaining 3 types of stains namely turmeric, oil and poster color has been also included.Click here for additional data file.


The most frequent ocular adnexal tumors and simulating lesions are lymphoproliferative disorders (LPDs), including malignant lymphomas and orbital inflammation with lymphoid hyperplasia or infiltration. IgG4-related orbital inflammation (IgG4-ROI) often involves lacrimal glands and other orbital tissues and is an important differential diagnosis. The present study evaluated clinical aspects of IgG4-ROI in a case series of orbital LPD. Sixty-two consecutive cases of orbital LPD, pathologically diagnosed from November, 2004, through March, 2011, were investigated. Histological types were 22 cases with MALT lymphoma, 11 cases with diffuse large B-cell lymphoma (DLBCL), 3 cases with other malignant lymphomas, 16 cases with IgG4-ROI, and 10 cases with non-IgG4-ROI. Ages of the IgG4-ROI group (56 ± 10 yrs) were significantly lower than the MALT lymphoma (71 ± 12 yrs) and DLBCL (75 ± 14 yrs) groups. Orbital lesions other than lacrimal glands were present in six cases including extraocular muscle swelling, mass lesions surrounding the optic nerve, and supraorbital and infraorbital nerves enlargements. Although none of the malignant lymphomas were related to IgG4, previous evidence suggested that malignant lymphomas can arise from IgG4-ROI. Based on this study (26%) and another report (33%), it is likely that nearly a quarter of orbital LPD are IgG4-ROI. The most frequent tumors and simulating lesions in ocular adnexa are lymphoproliferative disorders (LPDs), including malignant lymphomas and orbital inflammation with lymphoid hyperplasia or infiltration, some of which are historically called “orbital pseudotumors.” In Japanese studies, Goto  reportedWhen patients with suspected orbital LPD are encountered, tissue biopsy is preferred since image examinations alone cannot distinguish inflammatory lesions from malignant lymphomas. Pathological examination can also detect whether the lesion is related to IgG4 or not. IgG4-related disease (IgG4-RD) often involves lacrimal glands, which is now known as IgG4-related Mikulicz's disease , 6 or IgIn Kanazawa University Hospital in Japan, a 47-year-old woman with IgG4-immunopositive histopathology and an elevated serum IgG4 level of 1000 mg/dL was diagnosed as a first case of IgG4-related dacryoadenitis in November, 2004 . From thP = 0.00027 in a Student's t-test) and DLBCL  groups. There were no significant age differences between other combinations of the four groups.Histological types of the 62 orbital LPD were 22 cases (35%) with extranodal marginal zone lymphoma of mucosa-associated lymphoid-tissue type (MALT lymphoma), 11 cases (18%) with diffuse large B-cell lymphoma (DLBCL), 3 cases (5%) with other malignant lymphomas , 16 cases (26%) with IgG4-related orbital inflammation (IgG4-ROI), and 10 cases (16%) with non-IgG4-related orbital inflammation (non-IgG4-ROI) . None ofn = 15, normal range < 135 mg/dL) and an increased ratio of serum IgG4/IgG . In 9 patients, the serum IgE was also elevated. In all 16 cases, diagnosis was made by lacrimal gland biopsy. In pathology, these were characterized by lymphoplasmacytic infiltration, forming lymphoid follicles  and sclerosing fibrosis [Clinical data of sixteen cases of IgG4-ROI are summarized in Steroid therapy was performed for all the IgG4-ROI cases except three patients because their eyelid swellings decreased in size after biopsy surgery  and because of normal serum IgG4 (case number 1) and diabetes mellitus (case number 2). Eleven cases underwent oral prednisolone tapering therapy with an initial dose of 20, 30, or 40 mg per day. In two cases with diabetes mellitus, lower doses of oral prednisolone (10 mg daily in case number 10 and 8 mg daily in case number 15) were administered initially and reduced to a maintenance dose. All of the 11 cases essentially responded well to initial doses of oral prednisolone, but in 5 cases  ocular symptoms such as eyelid swelling deteriorated during tapering, and thus increase in the dosage was required. In case number 11 with retrobulbar mass , oral prIn this study, none of the malignant lymphoma cases was related to IgG4. However, some previous reports suggested that orbital malignant lymphomas can be related to IgG4. Cheuk reported three cases of ocular adnexal lymphoma (2 MALT lymphoma and 1 follicular lymphoma) arising in IgG4-related dacryoadenitis, and that the rate of transformation of malignant lymphoma in the background of IgG4-ROI was approximately 10% . Sato etThere is no doubt that IgG4-ROI most frequently involves the lacrimal gland, which is reported as IgG4-related dacryoadenitis and Mikulicz's disease –8. In adAmong 16 cases of IgG4-ROI, one patient (case number 1) presented normal levels of serum IgG4. In this case, the ratio of IgG4+/IgG+ cells in immunohistochemistry was around 30%, somewhat lower than values in diagnostic criteria , but IgGThe rate of IgG4-RD in 62 cases with orbital LPD was 26% in this study and was reported to be 33% out of 58 orbital LPD cases from another Japanese institute . Thus, i


Little information is available on antibiotic prescription management in German hospitals.The objective of this cross sectional study was to determine the prevalence and components of antibiotic stewardship (ABS) measures in German ICUs.A questionnaire survey was sent to all ICU-members of the German nosocomial infection surveillance system KISS (n=579) in October 2011. Data on antibiotic management structures were collected and analyzed by structural hospital and ICU factors.The questionnaire was completed by 355 German ICUs (61% response rate).The most common measures used (>80% of the ICUs) were personal restrictions for antibiotic prescriptions, routine access to unit-based bacterial resistance data and pharmacy reports on antibiotic costs and consumption. A small proportion of ICUs (14%) employed ABS consultants or infectious diseases specialists for the prescription of antimicrobial medication. Hospitals with their own integrated microbiological laboratory report twice as much to take part in surveillance networks of antimicrobial use (34%) and bacterial resistance (32%) compared with hospitals with external laboratories . Also, non-profit and public hospitals participate more often in surveillance systems for bacterial resistance than private hospitals .While the majority of ICUs report to have some antibiotic policies established, the contents and composition of these strategies vary markedly. Organizational-leveled control strategies to improve antibiotic control are already quite common in Germany. In contrast, other strategies widely considered effective such as the continuous surveillance of antimicrobial use and bacterial resistance or the employment of infectious disease consultants are still scarce. More effort is required with regard to incentives for infectious diseases consultants and to the participation in surveillance networks.This study provides a benchmark for future antibiotic stewardship programs.None declared


Prior antibiotic exposure has been associated with the emergence of antibiotic resistance in subsequent bacterial infections, whose outcomes are typically worse than similar infections with more antibiotic susceptible infections. The influence of prior antibiotic exposure on hospital length of stay (LOS) and costs in patients with severe sepsis or septic shock attributed to Gram-negative bacteremia has not been previously examined.A retrospective cohort study of hospitalized patients (January 2002-December 2007) was performed at Barnes-Jewish Hospital, a 1200-bed urban teaching hospital. Patients with Gram-negative bacteremia complicated by severe sepsis or septic shock had data abstraction from computerized medical records. We examined a consecutive cohort of 754 subjects .Escherichia coli (30.8%), Klebsiella pneumoniae (23.2%), and Pseudomonas aeruginosa (17.6%) were the most common organisms isolated from blood cultures. 310 patients (41.1%) had exposure to antimicrobial agents in the previous 90 days. Patients with recent antibiotic exposure had greater inappropriate initial antimicrobial therapy  and hospital mortality  compared to patients without recent antibiotic exposure. The unadjusted median LOS  following sepsis onset in patients with prior antimicrobial exposure was 13.0 days  compared to 8.0 days  in those without prior antimicrobial exposure (p < 0.001). In a Cox model controlling for multiple confounders, prior antibiotic exposure independently correlated with remaining hospitalized . Adjusting for potential confounders indicated that prior antibiotic exposure independently increased median attributable LOS by 5.0 days. Similarly, total hospital costs following sepsis onset was significantly greater among patients with prior antimicrobial exposure .Recent antibiotic exposure is associated with increased LOS and hospital costs in Gram-negative bacteremia complicated by severe sepsis or septic shock. Clinicians and hospital administrators should consider the potential impact of recent antibiotic exposure when formulating empiric treatment decisions for patients with serious infections attributed to Gram-negative bacteria. Staphylococcus aureus, enterococci and Gram-negative bacilli and increases in mortality, morbidity, length of hospitalization and cost of healthcare  v. $21,329 ; p < 0.001). LOS and total hospital costs following the onset of Gram-negative sepsis were strongly correlated .Figure th and 75th percentiles: $52,892; $134,310] v. $21,369 ; p < 0.001).When only hospital survivors were examined, subjects receiving prior antibiotic exposure had an unadjusted median increase of 7.0 days in LOS . Similarly, when only hospital survivors were examined total median hospital costs were significantly greater for patients with prior antibiotic exposure compared to patients without prior antibiotic exposure  associated with sepsis among patients having received prior antibiotic therapy. We previously demonstrated that prior antibiotic exposure was associated with a greater incidence of infection attributed to Gram-negative bacteria that were either antibiotic-resistant or multi-drug resistant . Prior aEscherichia coli and Klebsiella pneumoniae bloodstream infections in Oxfordshire, UK, over an 11 year period [Klebsiella pneumoniae  predicted a longer hospital LOS, as did age 7 years . On multivariate analysis, multidrug resistance remained a strong independent predictor of increased LOS. Similarly, Lautenbach et al. examined the longitudinal trends in prevalence of imipenem-resistant Pseudomonas aeruginosa (IRPA) from 2 centers from 1989 through 2006 [The findings from our study are consistent with those observed by other investigators. Webster et al. recently described resistance trends in r period . In theiugh 2006 . Isolatiugh 2006 ,13.Our study is unique in identifying prior antibiotic exposure as an independent risk factor for prolonged LOS among patients with Gram-negative bacteremia complicated by severe sepsis or septic shock. This observation suggests that clinicians should search for and identify the presence of prior antibiotic exposure as an important consideration when prescribing empiric antibiotic therapy to patients with severe sepsis or septic shock. The identification of prior antibiotic exposure should result in specific therapeutic interventions. For example, clinicians should avoid antibiotics or antibiotic classes to which the patient was previously exposed. Additionally, severity of illness appears to play an important role in determining which patients are at greatest risk for adverse outcomes from inappropriate antibiotic therapy . PatientThere are several important limitations of our study that should be noted. First, the study was performed at a single center and the results may not be generalizable to other institutions. However, the findings from other investigators corroborate the potential role of prior antibiotic exposure as an important determinant of outcome, including LOS, for patients with serious Gram-negative infections -7,24. SeIn conclusion, we observed that prior antibiotic exposure is relatively common and associated with adverse outcomes including greater hospital LOS among patients with Gram-negative bacteremia complicated by severe sepsis or septic shock. This observation suggests that clinicians treating patients with suspected Gram-negative bacteremia or sepsis should attempt to identify whether prior antibiotic exposure occurred. In clinical situations where recent antibiotic exposure is likely, but details concerning prior antibiotic exposure are unknown, combination empiric therapy directed against Gram-negative bacteria would be reasonable to increase the likelihood of appropriate therapy, until susceptibility data become available. Given the importance of prior antibiotic exposure as a risk factor for antibiotic resistance, inappropriate therapy, and increased mortality, and the availability of electronic medical records at many hospitals, institutions should try to formalize an approach for identifying prior antibiotic exposure in patients with serious infections.LOS: length of stay; ICD - 9 - CM: International Classification of Diseases, Ninth Revision, Clinical Modification; APACHE: Acute Physiology and Chronic Health Evaluation.Dr, Kollef's efforts were supported by the Barnes-Jewish Hospital Foundation. The authors have no other financial conflicts of interest or competing interests regarding this manuscript. Sharp & Dohme Corp. provided an unrestricted grant in support of this and other investigations. They had no role in the development or the conduct of this investigation.SM had full access to all of the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. SM contributed to the study conception and design, statistical analysis, and critical revision of the manuscript. MJ had full access to all of the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. MJ contributed to the study conception and design, acquisition of the data, statistical analysis, and drafting of the manuscript. RR contributed to acquisition of the data and critical revision for important intellectual content. MK had full access to all of the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. MK contributed to the study conception and design, statistical analysis, and critical revision of the manuscript. All authors read and approved the final manuscriptThe pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2334/12/56/prepub


AIDS-related Kaposi’s sarcoma (AIDS-KS) is the most common AIDS-associated malignancy and remains a significant cause of morbidity and mortality in sub-Saharan Africa. We describe the determinants of mortality among patients with AIDS-KS in a comprehensive HIV care and treatment program in Jos, Nigeria.We collected epidemiologic, clinical, staging and survival data for 357 patients with a diagnosis of AIDS-KS enrolling for HIV-care at the Jos University Teaching Hospital. Patients were staged according to the AIDS Clinical Trials Group (ACTG) criteria, which are based on the evaluation of tumor extension (T), CD4+ cell count (I), and patient’s systemic status (S), stratified by good (0) versus poor (1) risk. Information on survival was obtained through an active follow-up on verification of vital status of the patients. Survival analysis was computed by the Kaplan-Meier method, and the log-rank test was used to test the difference between subgroups.3), 77.5% had widespread tumor extension (T1) and 80.2% had systemic disease (S1). Poor immune system (I1) status  and presence of systemic disease (S1)  were independently associated with mortality.During the period of the study (2004-2008), there were 197 women (55.2%) and 160 (44.8%) men with AIDS-related Kaposi Sarcoma. Their mean age was 37±8 years and the median follow-up was 15 months (1-49 months). The median CD4+ and viral load were 107cells/mm³ and 58,561copies/ml respectively at baseline. Only 42 (11.8%) were on HAART at KS diagnosis, however all patients were commenced on HAART in line with existing national guidelines subsequently. 262(74.4%) had poor immune system status , 58% for S0 vs 49% for S1 (p=0.41), 60% for T0 vs 46% for T1 (p=0.19).Poor immune status and systemic disease are independent predictors of mortality in patients with AIDS-KS in Nigeria.