France implemented a comprehensive smoke-free law in two phases: Phase 1 (February 2007) banned smoking in workplaces, shopping centres, airports, train stations, hospitals, and schools; Phase 2 (January 2008) banned smoking in hospitality venues . This paper evaluates France’s smoke-free law based on the International Tobacco Control Policy Evaluation Project in France (the ITC France Project), which conducted a cohort survey of approximately 1,500 smokers and 500 non-smokers before the implementation of the laws (Wave 1) and two waves after the implementation (Waves 2 and 3). Results show that the smoke-free law led to a very significant and near-total elimination of observed smoking in key venues such as bars (from 94–97% to 4%) and restaurants (from 60–71% to 2–3%) at Wave 2, which was sustained four years later . The reduction in self-reported smoking by smoking respondents was nearly identical to the effects shown in observed smoking. Observed smoking in workplaces declined significantly after the law (from 41–48% to 18–20%), which continued to decline at Wave 3 (to 14–15%). Support for the smoke-free laws increased significantly after their implementation and continued to increase at Wave 3 . The findings demonstrate that smoke-free policies that are implemented in ways consistent with the Guidelines for Article 8 of the WHO Framework Convention on Tobacco Control (WHO FCTC) lead to substantial and sustained reductions in indoor smoking while also leading to high levels of support by the public. Moreover, contrary to arguments by opponents of smoke-free laws, smoking in the home did not increase after the law was implemented and prevalence of smoke-free homes among smokers increased from 23.2% before the law to 37.2% 5 years after the law. The public health harms associated with exposure to second-hand smoke have been well documented including premature death, lung cancer, heart disease, and coronary obstructive pulmonary disease (COPD) Smoke-free policies were implemented in two phases in France. Phase 1 was implemented in February 2007 in workplaces, shopping centres, airports, train stations, hospitals, and schools. In January 2008, the ban was extended in Phase 2 to hospitality venues . Media campaigns were held in France prior to the launch of each phase of the ban to inform the public about the forthcoming policies and to provide education about the health effects associated with exposure to second-hand smoke.The International Tobacco Control Policy Evaluation Project in France (the ITC France Project) was created in 2006 to evaluate France’s smoke-free legislation as well as other tobacco control policies implemented under the FCTC including health warning labels, price and taxation policies, tobacco advertising and promotion bans, cessation strategies, and education campaigns Three waves of a cohort survey of a nationally representative sample of smokers and non-smokers living in continental France have been completed between 2006 and 2012. The ITC France Wave 1 Survey was conducted just before the Phase 1 smoking ban was implemented in workplaces and other public venues in February 2007. The cohort was recontacted for the Wave 2 Survey after Phase 2 of the smoking ban. The Wave 3 Survey was conducted approximately 5 years after the ban in hospitality venues and 6 years after the workplace ban to provide an evaluation of the longer term impacts of the smoking ban and other policies.The effectiveness of the smoking ban in France has been reported in the literature based on ITC France Wave 1 (pre-ban) and Wave 2 (post-ban) data This paper reports the results of the ITC France Survey evaluation of the 2007–2008 smoking ban across a variety of indoor public venues at one pre-implementation and two post-implementation time points. We report the impact of the ban on observed smoking and self-reported smoking behaviour in workplaces, restaurants, and bars and pubs; public support for comprehensive smoking bans in several public places; and the prevalence of smoking restrictions at home.Prospective respondents were contacted via telephone (see below) and information was provided by the interviewer using a script built into the computer assisted telephone interview (CATI) program regarding the topic of the survey, the research institutions involved; prospective respondents were informed that all responses would be confidential. Respondents gave their consent verbally and the CATI script required the interviewer to enter the consent in the computer program before proceeding to the survey. As typical of telephone surveys, written consent was not obtained. However, prior to beginning the survey, the respondent was informed that he/she was free to decline to answer any question, and to terminate the survey at any time. The respondent was asked whether he/she could complete the survey at this time or at another more convenient time. All respondents were mailed compensation of a 10€ cheque (smokers for the 50-minute interview) or 8€ (non-smokers for the 30-minute interview) by mail after the survey interview (this was increased to 20€ at Wave 2 and Wave 3). Because of the cohort design, at Wave 2 and Wave 3, we recontacted all respondents from the previous wave by mail to inform them that they would be contacted by phone in the near future to invite them to participate in the next wave of the survey. When reached by phone, these recontact respondents were again read the information and consent script, and verbal consent was again obtained. The survey and research protocol was reviewed and cleared for ethics at each of the three survey waves by the Human Research Ethics Committee at the University of Waterloo, which waived the need for written informed consent from the participants.The ITC France Survey is a national longitudinal survey of adult (≥18 years of age) smokers and non-smokers. Smokers are those who have smoked more than 100 cigarettes in their life and smoke at least monthly. The CATI survey follows a random digit dialing (RDD) sampling design and covers continental France. To compensate for respondents lost to attrition, new respondents were randomly selected at Waves 2 and 3 using the same RDD sampling design and interview protocol as in Wave 1.14 The process for translation of the English survey into French was guided by the translation protocol described by International Agency for Research on Cancer (IARC) which calls for discussion of translation among bilingual persons knowledgeable about the survey content.12 The ITC France team researchers in Paris met this qualification.The questions used in the ITC France Survey were all adapted from the conceptual model and questionnaire of the ITC Four Country Survey.Wave 1 of the ITC France Survey was conducted by the survey firm Atoo between December 2006 and February 2007 with a cooperation rate of 75.3%. Wave 2 was conducted from September to November 2008 by the Institut de sondage Lavialle (ISL). Wave 3 was conducted from September to December 2012 by the firm BVA. The complete survey methodology can be found on the ITC Project website Observed smoking in bars and restaurants was measured among those who had visited in the last 6 months with the questions “The last time you visited, were people smoking inside the café, bar or pub/restaurant?”. The response options were “yes/no”. Smoking behaviour in bars and restaurants was assessed with the question “Did you smoke at all at the bar/restaurant during your last visit, either indoors or outdoors?” Respondents who answered “yes” were asked whether they smoked inside the bar/restaurant, outside, or both.Observed smoking in workplaces was measured among those who were employed outside the home in indoor workplaces with the question “In the last 30 days, have people smoked in indoor areas where you work?”. The response options were “yes/no”. Smoking behaviour in workplaces was measured with the question “In the last 30 days, have you smoked in indoor areas at work?” (yes/no).Support for smoke-free law in bars and restaurants was assessed with the question “Do you support or oppose the French total ban on smoking inside cafés, bars and pubs/restaurants?” with options “strongly support/support/oppose/strongly oppose”. Support for a smoke-free law in cars with children was assessed with the question “Do you support or oppose a French total ban on smoking in cars with children with them?” with the same response options. Support for bans in other places was assessed with the question “For each of the following public places, please tell me if you think smoking should be allowed in all indoor areas, only in some indoor areas, or not allowed indoors at all.” Overall support for the smoking ban was measured with “Overall, would you say that the ban on smoking in restaurants, cafés and bars, and other public places is a good thing or a bad thing?” (very bad/bad/good/very good). Support for a ban in outdoor eating areas of restaurants was measured with the question “Do you think that smoking should be allowed in all outdoor eating areas, in designated outdoor eating areas such as smoking terraces, or not allowed in outdoor eating areas at all?” (yes/no). Support for a ban on smoking in cars with children was measured with “Do you support or oppose a French total ban on smoking in cars with children in them?”(strongly support/support/oppose/strongly oppose). Home smoking bans were measured with “Which of the following best describes smoking inside your home?” .Sampling weights were rescaled to population totals based on the prevalence of smoking in France. These inflated weights were then rescaled to the total number of smokers and non-smokers in the sample. Logistic regression models for longitudinal data  were used to analyze differences in the measure of interest across waves. An unstructured within-group correlation matrix was used in the modeling to account for within-subject correlation. All of the models simultaneously controlled for gender, age at recruitment , wave, and smoking status (smoker vs. non-smoker). It has been documented that respondents who are newly recruited may vary in their responses compared to those who have completed one prior wave, who may vary from those who have completed two prior waves, and so on A total of 2260 (1735 smokers and 525 non-smokers) respondents completed Wave 1 of the ITC France Survey. A total of 2219  respondents completed Wave 2. Of these 2219 respondents, 1231 were from the cohort smoker sample and 414 were from the cohort non-smoker sample, resulting in an overall retention rate of 72.8%. The Wave 2 replenishment sample consisted of 473 smokers and 101 non-smokers. The cooperation rate among the Wave 2 replenishment sample was 80.5%. At Wave 3, a total of 2204  completed the ITC France Survey. Of these 2204 respondents, 1198 were from the cohort smoker sample and 390 were from the cohort non-smoker sample, which included 17 respondents who were non-smokers at Wave 2 and became smokers at Wave 3. There were 502 smokers and 114 non-smokers in the Wave 3 replenishment sample. The cooperation rate among the Wave 3 replenishment sample was 80.7%.The overall retention rate at Wave 3 was 71.6%. Note that 95% confidence intervals are presented in parentheses throughout the results.Among smokers, observed smoking in restaurants declined from 70.8% (63.1–77.5) before the law (Wave 1) to 2.0% (1.2–3.4) at first post-ban survey (Wave 2)reported by smokers and reported by non-smokers), 8 months after the law was implemented (p<.001). Observed smoking in restaurants among non-smokers also declined from 60.4% (50.8–69.3) at Wave 1 to 3.0% (1.4–6.2) at Wave 2 (p<.001). This very low prevalence of restaurants with smoking was sustained at the second post-ban survey (Wave 3) to 0.7% (0.4–1.4) among smokers and 2.1% (0.9–4.8) among non-smokers); this decline among smokers (from 2.0% (1.2–3.4) to 0.7% (0.4–1.4)) was statistically significant (p = .040). The slight decline from 3.0% (1.4–6.2) to 2.1% (0.9–4.8) among non-smokers was not statistically significant (p = .550).These findings are presented in We also examined self-reported smoking behaviour in restaurants. At Wave 1, 76.0% (67.4–82.9) of smokers smoked indoors or indoors and outdoors in a restaurant at last visit. This percentage decreased to 1.4% (0.7–3.0) at Wave 2 (p<.001) and was sustained at Wave 3 to 1.8% (1.0–3.4) (p = .622).Observed smoking in bars declined from 96.6% (95.1–97.7) at Wave 1 to only 3.5% (2.4–5.0) at Wave 2 among smokers (p<.001). Among non-smokers, observed smoking in bars also declined from 93.5% (90.0–95.8) at Wave 1 to 4.4% (2.5–7.6) at Wave 2 (p<.001). The prevalence of observed smoking in bars at Wave 3 slightly increased to 6.4% (4.9–8.4) for smokers and 7.9% (4.8–12.7) for non-smokers. This slight increase was significant among smokers (p = 0.024), but not significant among non-smokers (p = .161).These findings are presented in Our analysis of self-reported smoking in bars found that at Wave 1, 95.5% (93.4–96.9) of smokers smoked inside or inside and outside a bar at last visit. This percentage decreased to 1.8% (1.0–3.2) at Wave 2 (p<.001). At Wave 3, 6.4% (4.5–9.2) of smokers reported smoking inside or outside a bar at last visit. This was significantly different from Wave 2 (p<.001) and Wave 1 (p<.001).Observed smoking in workplaces also declined significantly from 47.8% (42.9–52.8) for smokers and 41.1% (33.9–48.7) for non-smokers at Wave 1 to 20.0% (16.9–23.6) and 17.9% (13.2–23.9), respectively, at Wave 2 (p<.001 for both smokers and non-smokers). There was a further decline at Wave 3 to 14.5% (11.7–17.7) among smokers and 13.5% (9.1–19.6) among non-smokers, however the slight decrease was not significant for smokers (p = .056) and non-smokers (p = .340). The sample used in the analysis of observed smoking in the workplace included respondents who reported that smoking is never allowed in any indoor area, smoking is allowed in some inside areas , and smoking is allowed in any indoor area. The presence of designated smoking rooms is a possible confounder, thus we also examined observed smoking in workplaces among those respondents who reported that smoking was not allowed in any indoor area of their workplace. For these workplaces, observed smoking also declined significantly from 18.6% (14.3–24.0) for smokers and 21.6% (14.2–31.5) for non-smokers at Wave 1 to 12.0% (9.5–15.0) and 9.7% (6.1–15.0), respectively, at Wave 2 . The slight decline at Wave 3 to 6.7% (4.9–9.1) among smokers was significant (p = .019), however the decline to 6.0% (3.2–10.7) among non-smokers was not significant (p = .352). These findings are presented in The analysis of self-reported smoking indoors in workplaces showed a similar trend. At Wave 1 39.6% (34.5–45.0) of smokers who worked outside the home in indoor workplaces smoked indoors regardless of their workplace smoking policy. This decreased to 14.1% (11.2–17.6) at Wave 2 (p<.001) and 11.8% (9.1–15.2) (p = .339) at Wave 3. Among those who reported a complete indoor workplace smoking ban at each wave, indoor smoking prevalence was low at Wave 1 (7.0% (4.5–10.8)) and decreased further at Wave 2 (4.6% (3.1–6.9)) and Wave 3 (3.3% (2.0–5.4)). However, the reductions at Wave 3 were not significant compared to Wave 1 (p = .104).Support for the smoke-free law in restaurants increased significantly after the law was implemented from 79.5% (74.9–83.5) to 89.8% (86.0–92.6) for non-smokers (p<.001) and from 52.8% (48.3–57.1) to 77.4% (74.2–80.3) for smokers (p<.001), with support continuing to increase at second post-ban follow-up for smokers (87.9% (85.4–90.0); p<.001) and for non-smokers (94.0% (90.1–96.4); p = .044). The pattern of the increase in support for smokers and non-smokers is presented in As shown in At Wave 1, about 58.6% (54.1–62.9) of smokers said that the ban on smoking in public places was “a good thing” or “a very good thing” compared to 83.7% (79.2–87.4) of non-smokers (p<.001). At the first post-ban wave, these percentages were significantly higher for smokers (87.7% (85.6–89.5), p<.001) and for non-smokers (95.7% (93.6–97.2), p<.001). Support stayed high at the second post-ban follow-up for smokers (89.4% (87.2–91.3)) and non-smokers (96.9% (94.5–98.2)).The ITC France Survey at Wave 2 and Wave 3 included measures assessing support for possible future smoke-free initiatives in two areas: outdoor eating areas and smoking in cars with children see .Support for a ban on smoking in outdoor eating areas remained stable between Wave 2 (36.6% (33.3–40.2) for smokers; 32.2% (27.1–37.7) for non-smokers) and Wave 3 (35.4% (31.4–39.7) for smokers; 35.2% (29.5–41.5) for non-smokers).The overwhelming majority of smokers and non-smokers at Wave 3 would support a smoking ban in cars with children (89.2% (86.8–91.1) vs. 92.2% (89.2–94.5), p = .385). There has been no change over time in this high level of support.An argument sometimes made against smoke-free laws is that such laws would displace smoking from public places like bars/pubs into the home. If this were true, then this would represent an adverse consequence of such laws. We tested this possibility over the three waves of the ITC France Survey. As shown in The findings reported from three waves of the ITC France Survey over six years demonstrate clearly that the 2007–2008 smoke-free law led to near-total elimination of smoking indoors in key public venues, including restaurants and bars and considerable reductions in smoking in indoor workplaces. The percentage of bars and restaurants where there was observed smoking decreased from near 100% for bars and about 60–71% for restaurants to about 4% for bars and 2–3% for restaurants at the first post-ban measure. Even 5 years after implementation of the ban in hospitality venues, the percentage of observed smoking in restaurants was about 1–2% and smoking in bars was just 6–8%. Similar reductions were evident in self-reported smoking in these venues.The near total elimination of smoking in these public places where before there was very high or near universal levels of smoking indicate the power of smoke-free laws when they are comprehensive and implemented properly. Previous ITC comparative evaluation of comprehensive vs. partial workplace smoke-free legislation in Ireland, England, and the Netherlands provides evidence that comprehensive smoke-free legislation as implemented in Ireland and England had positive effects on increasing quit attempts and increasing quit success respectively, while partial smoke-free legislation in the Netherlands was not shown to have an impact on quit attempts or quit success The data on support for smoking bans show that support for the law increases substantially after its implementation, with 77% (74.0–80.2) of smokers supporting the ban in bars, pubs, and cafés and over 87% (85.4–90.0) of smokers supporting the ban in restaurants, slightly less than non-smokers (94% (90.1–96.4)). These findings add to the body of evidence across multiple countries that support for smoking bans in public places increases, especially among smokers, after the implementation of the ban The data also confirm that smoking bans in public places do not lead to displacement of smoking into the home , were people smoking inside the restaurant?) or were those that can only be ascertained through self-report . Another potential limitation would be in biases due to attrition. However, there were no significant differences in the Wave 1 demographic characteristics  and key smoking variables (CPD and ever tried to quit) among those who were lost to attrition at Wave 2 and those who completed the Wave 2 Survey. Similarly, there were no significant differences in the Wave 1 responses for noticing smoking, support for bans, and smoking inside bars/restaurants/workplaces among those respondents who participated in Wave 2 and those who did not.The results of this longitudinal evaluation study demonstrate the substantial success of France’s comprehensive smoke-free laws, both at one year and five years after the law. Future waves of the ITC France Survey will continue to evaluate support for and compliance with smoke-free laws, as well as long term effects of the law on smoking cessation. The ITC France Survey will also continue its rigorous evaluation of France’s policies in other domains of the FCTC, for example, price/taxation, and health warning labels. Implementing stronger tobacco control policies across the range of the FCTC will be required to combat the recent increase in smoking in France


The published evidence concerning the safety, efficacy, and patient satisfaction for implantation of the current model of the artificial urinary sphincter (AS 800) in men with post prostatectomy urinary incontinence was the objective of this review. A Pub Med English language literature search from 1995 to 2011 was performed. A majority of men who undergo AUS implantation for post prostatectomy urinary incontinence achieve satisfactory results (0 to 1 pad per day). Infection rates range from 0.46 to 7%, cuff erosion rates range from 3.8 to 10%, and urethral atrophy ranges from 9.6 to 11.4%. Kaplan-Meier 5 year projections for freedom from any reoperation were 50% for a small series and 79.4% for a larger series. Kaplan-Meier projections for freedom from mechanical failure were 79% at 5 years and 72% at 10 years. In another series 10 year projections for freedom from mechanical failure were 64%. Although the artificial urinary sphincter (AUS) is the gold standard for the treatment of this disorder, most men will continue to need at least one pad per day for protection, and they are subject to a significant chance of future AUS revision or replacement. There is a wide range in the reported incidence of postradical prostatectomy urinary incontinence coming from individual series presumably due to inconsistent definitions of incontinence and differing modes of assessment. When large populations of postradical prostatectomy patients are surveyed, however, a more consistent pattern is observed. In a series of 1291 postprostatectomy patients, significant urinary incontinence persisted in 8.4% of men at eighteen months .The artificial urinary sphincter (AUS) is widely regarded as the gold standard for the treatment of post prostatectomy urinary incontinence . This prSince 1983 the basic design of the AUS has been unchanged; however, there have been numerous modifications to device components leading to both increased continence and longer device life. These component changes include narrow back cuffs for bulbous urethral use, smaller (3.5 and 4.0 cm) cuffs, surface-coated cuffs to reduce wear, kink-resistant tubing, tubing sleeves to reduce wear, and sutureless connectors to facilitate making connections and to reduce connector failures.A PubMed English language literature search from 1995 through 2011 for keywords: artificial sphincter; urinary sphincter, artificial; prosthesis failure; prostatectomy/ae [adverse effects]; patient satisfaction; quality of life was performed. Thirteen articles were found where data relevant to patients with post prostatectomy incontinence could be separated from other AUS uses. These articles were examined to perform this paper.There is no standardization for reporting pre- and post-AUS levels of incontinence. In a study of 50 patients with a median followup of 23.4 months, the preoperative levels of incontinence were such that 70% wore an average of 6 diapers a day, and 24% wore an average of 7.4 pads per day [In a study of 54 men with mean follow-up of 7.2 years, 54% were socially continent (0 to 1 pad per day). Mean pad score before AUS implantation was 2.75, and it decreased to 0.97 after AUS implantation .In a group of 113 patients with mean follow-up of 73 months (range 20 to 170), 32% were pad free, 33% used 1 pad per day, 14% used 2 pads per day, 17% used 3 pads per day, and 5% used more than 3 pads per day .In a study of 33 men aged 75 years or greater with an average follow-up of 5 years, mean pad use decreased from 6.7 pads per day range 3–10) to 0.8 pads per day (range 0–2) [ to 0.8 pIn a study comparing 435 first time AUS implants to 119 repeat AUS implants, pad use of 0 to 1 day was noted in 90% of the primary implants and 82% of the secondary implants [In 124 patients with a median follow-up of 6.8 years, 27.1% required no pads and 52.0% required 1 pad per day after AUS implantation [In a cohort of 71 men with mean follow-up of 7.7 years (range 0.5–16), 27% used no pads, 32% used 1 pad, 15% used 1–3 pads, and 25% used more than 3 pads daily .In a series of 40 men with mean follow-up of 53.4 ± 21.4 months (range 27–132), pad count decreased from 4.0 ± 0.9 to 0.62 ± 1.07 per day, and on a visual analog scale the impact of the incontinence decreased from 5.0 ± 0.7 to 1.4 ± 0.93 .Cuff erosion  occurrinAUS infection  occurrinWhen men achieve a satisfactory level of continence following activation of their AUS (0 to 1 pad per day) and later a gradual increase in pad use occurs, an AUS trouble shooting protocol should be employed . If otheUrethral atrophy rates are shown in For penile prosthesis implantation the American Urological Association Erectile Guidelines Committee has recommended that freedom from mechanical failure be reported in terms of Kaplan-Meier projections . This alKaplan-Meier reporting was used in one series where 66 AUS implantation patients were available for a mean follow-up of 41 months, The 5-year Kaplan-Meier projections for freedom from any reoperation was 50% and for freedom from any cuff revision was 60% . In anotMechanical failure rates for the remaining studies are shown in In one series, 58 men with AUS and no prior radiation (group 1) were compared to 28 with AUS after radiation therapy for prostate cancer (group 2) . Mean foIn another study, 76 men with AUS implantation and no radiation were compared to 22 men with AUS implantation after radiation . UrethraIdeal satisfaction studies would be prospective and used standardized questionnaires administered pre- and post-operatively. In addition since incontinence may have a significant quality-of-life impact on the partner, including partners in these studies would be desirable. Unfortunately, nearly all studies are retrospective and use a variety of nonvalidated satisfaction scales.In a study of 50 patients with a median follow-up of 23.4 months, 90% reported satisfaction, 96% would recommend AUS implantation to a friend, and 92% would have the AUS placed again .In a study of 54 men with mean follow-up of 7.2 years, subjective improvement was 4.1, and overall satisfaction was 3.9  .In a group of 113 patients with mean follow-up of 73 months (range 20 to 170), 28% were very satisfied, 45% were satisfied, 18% were neutral, 6% were dissatisfied, and 4% were very dissatisfied .In 71 men with a mean follow-up of 7.7 years (range 0.5–16), 58% were very satisfied, 19% were satisfied, and 23% were unsatisfied . Significant urinary incontinence following radical prostatectomy which persists for more than one year occurs in as many as 8% of men. Although the AUS is the gold standard for the treatment of this disorder, most men will continue to need at least one pad per day for protection, and they are subject to a significant chance of future AUS revision or replacement.Current reporting of AUS implantation results is far from ideal. Freedom from mechanical failure and other complications should be reported in terms of Kaplan-Meier projections. Quality-of-life and patient satisfaction studies should be prospective, include partners, and use validated questionnaires.


Bounce was defined as an increase ≥ 0.2 ng/ml above the nadir, followed by a spontaneous return to the nadir. Biochemical failure (BF) was defined using the criteria of the Phoenix conference: nadir +2 ng/ml.Men treated with exclusive permanent 198 men were included. After a median follow-up of 63.9 months, 21 patients experienced a BF, and 35.9% had at least one bounce which occurred after a median period of 17 months after implantation (4-50). Bounce amplitude was 0.6 ng/ml (0.2-5.1), and duration was 13.6 months (4.0-44.9). In 12.5%, bounce magnitude exceeded the threshold defining BF. Age at the time of treatment and high PSA level assessed at 6 weeks were significantly correlated with bounce but not with BF. Bounce patients had a higher BF free survival than the others . In case of PSA increase, PSA doubling time and velocity were not significantly different between bounce and BF patients. Bounces occurred significantly earlier than relapses and than nadir + 0.2 ng/ml in BF patients .High PSA value assessed 6 weeks after brachytherapy and young age were significantly associated to a higher risk of bounces but not to BF. Long delays between brachytherapy and PSA increase are more indicative of BF. Permanent seed prostate brachytherapy has become a standard treatment for localized prostate cancer ,2. The fThe main objectives of this study were to identify predictive factors of bounce occurrence and criteria distinguishing bounces from true biochemical relapses.Prostate brachytherapy with permanent iodine seeds was first used at the Centre Léon Bérard in November 1999 and so far more than 500 patients have been treated. In order to investigate PSA bounces and obtain a significant follow-up, our study population included all the men for prostate carcinoma classified in the low or intermediate group according to D'Amico et al. classification [Two different brachytherapy techniques were consecutively used: a free seeds technique with use of the Mick applicator during the first 3 years, then the "FIRST" technique from Nucletron  characterized by the use of a seed projector. During both periods, the prescribed dose was 160 Gy to the entire prostate (applying TG43 guidelines) and an intra-operative dosimetry was performed based on ultrasound delineation of the prostate and organs at risk. PSA testing were performed 6 weeks and 6 months after the brachytherapy, then every 6 months up to 5 years, and at least once a year thereafter. The frequency of PSA testing was usually increased to every 3 months for patients who experienced a PSA increase.As suggested by the majority of authors, PSA bounce was defined as an increase of at least 0.2 ng/ml above the nadir, followed by a spontaneous decrease to or below pre-bounce level -22. An iThe duration of the bounce was defined as the time from the pre-bounce nadir to the first PSA level below this nadir. The magnitude of the bounce was defined as the difference between the nadir and the highest value of the peak. Time to onset was assessed by delay between brachytherapy and date corresponding to the first date where PSA increased by more than 0.2.Biochemical relapse was defined according to Phoenix criteria (PSA nadir + 2 ng/ml). True biochemical relapse was defined as a PSA increase fulfilling Phoenix criteria but not bounce definition (spontaneous PSA decrease) or post-brachytherapy positive biopsy or start of a salvage treatment. In case of patients with a bounce and then a true biochemical relapse, data were treated in the analysis of bounce and relapse. In the case of patients having two or more bounces, only the first one was used for analysis.PSA velocity (PSAV) and PSA doubling time (PSADT) were calculated with formula: PSAV = (PSAf - PSAn)/ΔT and PSADT = ln(2) ΔT/(ln(PSAf) - ln(PSAn)) where ΔT is the delay between PSAn (nadir) and PSAf . Date of true biochemical failure was defined as the delay between brachytherapy and the first date where PSA is higher than 2 ng/ml from the nadir, the date of post-brachytherapy positive biopsy or the date of start of a salvage therapy.True biochemical relapse-free survival delay was calculated from the date of the brachytherapy to the date of relapse. The probability of true biochemical relapse-free survival was calculated using the Kaplan-Meier method.Clinical or dosimetric factors possibly predictive of bounces were assessed by logistic regression. Factors included in the univariate analysis were: patient age, tumour risk group according to D'Amico et al. classification (low vs. intermediate), pre-BT PSA, PSA value assessed 6 weeks after the brachytherapy, brachytherapy technique (free seeds vs. FIRST) and intra-operative dosimetric parameters .Potential predictive factors of bounces with a 0.1 significance level in univariate analysis were included in a multivariate logistic regression.These factors were also tested to predict presence of true biochemical relapse with a logistic regression model.Bounce patients were compared to patients with true biochemical relapse: PSAV, PSADT, nadir before rise and time to onset were tested with a wilcoxon test. In case of patients with bounce and relapse, they were considered only with bounce for this comparison.All statistical analyses were done using SAS software v.9.1 for Microsoft Windows .198 men fulfilled the inclusion criteria and were included in the study. Their characteristics are detailed in Table The median follow-up was 63.9 months (range 36 - 119.4), 88.4 months (range 36.3 - 95.6) for the former implantation technique and 52.9 months (range 36 - 81.9) for FIRST.A total of 2,219 PSA values were recorded, with a median number of 10.5 per patient (range: 3 - 24). Figure At the first visit, scheduled 6 weeks after the brachytherapy , 20% of the patients had a PSA increase from its initial value . This value was not taken into account in bounce identification as it may have been altered by implantation and prostate edema.At the time of analysis, 80.3% of the patients had achieved a nadir < 0.5 ng/ml.Seventy one patients (35.9%) experienced at least one bounce defined by a PSA increase of at least 0.2 ng/ml followed by a spontaneous decrease to or below pre-bounce level. Ten patients experienced 2 bounces and one patient experienced 3 bounces. The respective proportion of bounces defined as a PSA increase with a threshold of 0.1, 0.4 and 2 ng/ml followed by a decrease to or below pre-bounce level were 48.5, 25.8 and 4.5% respectively. Characteristics of the bounces are presented in Table Median time to onset was 17 months (3.6-50.2), and 56.3% of the bounces occurred between 12 to 24 months after the brachytherapy. After 30 months, bounces were rare (8.5%).The median bounce duration was 13.6 months (range: 4.0-44.9), and 75% were limited to 20 months. 18.3% lasted over 2 years. The median increasing part duration of the bounces, which is the most agonizing, was 6.4 months (1.3-23), and was limited in 75% of the bounces to 11 months.The median magnitude was 0.6 ng/ml (0.2-5.1). It was lower than 1 ng/ml in 72% of the cases, but higher than 2 ng/ml in 12.5% of the bounces (9 patients).In univariate analysis, younger age was significantly associated with a higher probability of a bounce occurrence p = 0.003). There was also a trend for higher PSA value at 6 weeks (p = 0.053)  and higher PSA value at 6 weeks  were significantly associated with a higher probability of a bounce Table . PatientStrictly applying the RTOG-ASTRO Phoenix consensus definition, 21 patients (10.6%) were considered to have a biochemical relapse (nadir +2 ng/ml), resulting in a 4 year relapse free survival (RFS) of 92% (CI95% = 87-95). Six patients had salvage hormonal therapy (3 with positive biopsies), including 2 who did not meet the BF criteria (nadir +2 ng/ml).In 9 cases, the PSA raise overcame the threshold of 2 ng/ml, but was followed by a spontaneous decrease to or below pre-bounce level fulfilling bounce definition. Thus, the "true recurrence rate"  was 7.1%  and a 4 year RFS of 95% (CI95% = 91-97)."True recurrences" (definition above) occurred respectively in 5 pts belonging to the intermediate risk group (8.8%) and in 9 pts to the low risk group (7.1%) (p = 0.686).In univariate analysis, no factor was found being predictive of true relapse. Results are shown in Table There was a better true biochemical relapse free survival in patients who experienced a bounce .Of the 71 patients who experienced a bounce, only one subsequently had a true biochemical relapse; he belonged to the intermediate prognosis group. Hence, none of the patients in the low risk prognosis group who had a bounce experienced a true biochemical relapse. Moreover, none of the patients who experienced a bounce > 2 ng/ml experienced a true biochemical recurrence.One patient experienced a bounce followed by a true biochemical relapse. He was considered as bounce patient here. Table Median PSADT was 11.5 months for the 71 patients who experienced a bounce (range: 1.0-70.2) and 12.0 months for the 14 patients with a true recurrence without bounce (range: 3.9-49.5). The median PSA velocity was estimated at 0.11 ng/ml/month for patients experiencing bounce (range: 0.01-0.85) and 0.12 ng/ml/month for patients with true relapse (range: 0.03-0.58). There was no difference between groups.The median nadir before the rise was 1.38 ng/ml (range: 0.02-6.6) for bounce patients and 1.00 (range: 0.04-3.04) for true relapse patients (p = 0.25).Bounces occurred significantly earlier than true BF . Furthermore, the level of nadir + 0.2 ng/ml was reached significantly later in true BF cases than in bounce: 27.8 versus 17 months (p < 0.0001)125I seeds. They attempted to correlate the topography of the initial disease with the metabolic activity during the bounce and with the exact locations of local relapses in 24 patients [The long-known phenomenon of PSA bounce after prostate brachytherapy can lead to patient anxiety, unnecessary imaging exams or prostate biopsies, and even to inappropriate administration of salvage therapy. So far, the bounce mechanisms are not understood and their etiology unknown. It has been hypothetized to be linked to inflammation, radiation prostatitis or vascular fibrosis. Recently Kirilova et al. published prostate 3D MR spectroscopic assessments during PSA bounce in patients treated with permanent patients . They obPSA measurement has been shown to be a reliable method for monitoring but there is currently no consensual definition of a bounce. For most authors, bounces correspond to any variation beyond a given threshold above the PSA nadir, usually +0.2 ng/ml -22. AlteAge is the most commonly reported predictive factor for a bounce Table . It seemProstate volume (> 35 ml) has been identified as predictive of bounce by Stock et al. , but this correlation has not been observed by the other authors . The traDas et al. tried to correlate PSA bounce to various events. They reported that 23% of the bounces may be subsequent to ejaculation, cycling, invasive exams, or radiation proctitis,  but thosIntra-operative and post-brachytherapy dosimetric factors have also been largely studied. Stock et al. found that D90 > 160 Gy was predictive of bounce  . ToledanPre-brachytherapy PSA value has not been correlated with the occurrence of a bounce, except in Makarewicz's HDR brachytherapy experience. In this report, the investigators showed that patients who experienced a bounce had greater pre treatment PSA value than the others  .Nevertheless, Merrick et al. have reported a correlation between a first high post treatment PSA value and the occurrence of a bounce  , but in 125 I permanent implantation. Merrick stated that the use of 103 Pd lead to a half likelihood of bounce  [125 I and 103 Pd led by Bostancic et al. They have shown by multivariate analysis that 125 I is significantly associated with a higher frequency of bounce in hormone-naive patients (45.7% with 125 I vs. 14% with 103 Pd) and in patients receiving neoadjuvant hormonal deprivation (respectively 28.1 and 20.7%) [Most of the studies were based on = 0.002) . Those rd 20.7%) . The dosd 20.7%) .PSA bounce phenomenon and hormonal therapy is more confusing as the spike could be a consequence of the end of hormonal deprivation and of the testosterone recovery. For Patel and Toledano, ADT had no influence either on bounce rate or its magnitude ,32. SimiIt has been hypothesized that PSA bounce after brachytherapy could be predictive of biochemical control, whereas it is known to be correlated with biochemical failure after EBRT. In a series of 4,838 patients treated with EBRT, Horwitz et al. observed a bounce (defined as a ≥ 0.4 ng/ml PSA increase) in 20% of cases, and this bounce was independently correlated to biochemical failure . These rPatel et al. analyzed a series of 295 patients treated with brachytherapy , with quite a short median follow up of 38 months. They observed that the BF-free survival assessed using the ASTRO consensus was 100% in the bounce group (28% of the population) vs. 92% in other patients (p = 0.018) . In an oIn our series, we observed only one biochemical relapse after a PSA bounce in one patient who belonged to the intermediate prognosis group. With 64 months follow-up, the occurrence of a bounce was therefore statistically correlated to biochemical disease-free survival in the subgroup of patients with "favorable prognosis" (p = 0.039).Distinguishing a benign PSA bounce from genuine biochemical recurrences is a major issue. On one hand it would reassure most patients with a PSA raise, and on the other hand, permit detection of true relapses in order to avoid expensive investigations such as 18-F choline PET/CT or invasive biopsies. To date, only follow up permits distinction of bounce from true BF. As illustrated in this series, 4 patients experienced an increase of the PSA over +2 ng/ml, and had begun a spontaneous decrease of their PSA at the time of analysis (of more than 1 ng/ml for 2 of them), but still could not be classified as bounces as the PSA did not return to the nadir.Despite these limitations, Phoenix criteria have appeared to be more accurate than the ASTRO criteria to predict clinical outcomes in prostate cancer patients treated with either EBRT or BT .Kuban et al. led a comparison of 12 different BF definitions on a large series of 2,693 men treated with permanent seed brachytherapy. They concluded that nadir +2 ng/mg provides the best sensitivity/specificity balance (70% and 89% respectively) . PicklesIn order to take into account the lack of specificity of the Phoenix criteria, several definitions have been proposed. Patel et al. have simply suggested that a bounce should never exceed the pretreatment PSA level . This paIn an attempt to differentiate bounce from true biochemical relapse, several parameters have been tested. Mitchell et al. reported a series of 205 patients, and observed 79 bounces defined as an increase of ≥ 0.2 from the nadir followed by spontaneous decrease to the nadir value or under, and 6 Phoenix true biochemical relapses. They found that PSA velocity was 0.08 ng/ml/month for bounces versus 0.28 ng/ml/month for true Phoenix biochemical relapses (p = 0.0005). Using the former ASTRO criteria, they did not observe any significant differences. The authors failed to demonstrate a predictive threshold for PSADT . PSA velTime to onset of PSA increase has been shown to be useful to distinguish bounce and relapse. Merrick et al. reported that 83% of the bounces occurred in the first 30 months following brachytherapy . Ciezki The authors declare that they have no competing interests.Presented at the annual ASTRO meeting held in San Diego (31/10 - 4/11/2010).PP and RM coordinated the study. RM: data collection. AB, RM: statistical analysis. RM, PP, AB: manuscript preparation. All authors read and approved the final manuscript.


This document is the result of a study of 1 November 2010 to 30 March 2011 on nosocomial infections at the Maternity Issaka Gazobi.This prospective survey covered a total of 139 patients of Obstetrics.After analyzing the data, it appears that: The prevalence of nosocomial infections was 7.2%, including 0.7% of endometritis, urinary tract infection 0.7% and 5.8% of surgical site infection distributed as follows: 3.5% and 2.2% of deep and superficial infections respectively. Nearly eight out of 10 patients were from the urban community of Niamey and 70% of infected patients are housewives. Prolonged urinary catheterization, as well as antibiotic misuse were the main contributing factors. In newborns, the prevalence of nosocomial infections was 5% of which 1.4% were skin infections and 3.6% eye infections, no umbilical cord infection was recorded. The mortality rate of newborns was 14.4%. Nosocomial infections are and remain a public health problem.This is what we contribute to formulate recommendations for the administrative and health authorities and for healthcare workers.None declared


C. monnieri fruit, has been shown to suppress tumor migration and invasion. However, the effects of osthole in human GBM are largely unknown. Focal adhesion kinase (FAK) is important for the metastasis of cancer cells. Results from this study show that osthole can not only induce cell death but also inhibit phosphorylation of FAK in human GBM cells. Results from this study show that incubating GBM cells with osthole reduces matrix metalloproteinase (MMP)-13 expression and cell motility, as assessed by cell transwell and wound healing assays. This study also provides evidence supporting the potential of osthole in reducing FAK activation, MMP-13 expression, and cell motility in human GBM cells.Glioblastoma multiforme (GBM) is the most common type of primary and malignant tumor occurring in the adult central nervous system. GBM often invades surrounding regions of the brain during its early stages, making successful treatment difficult. Osthole, an active constituent isolated from the dried  Tumor metastasis is a highly complex multistep process that includes changes in cell-cell adhesion properties . GlioblaInvasion of glioma cells into adjacent brain structures occurs through the activation of a variety of molecules including the matrix metalloproteinases (MMPs), which play an important role in tumor invasion . MMPs caCnidium monnieri (L.) Cusson, is represented in many Mediterranean and Middle-east regions plants and has been shown to exert a wide array of biological effects, such as contractility-based motility of different cells and tissues [Frequent consumption of natural fruits and vegetables has been considered with reducing risk of developing various of cancer and mortality ,26. Osth tissues . Osthole tissues , anti-os tissues , and ant tissues  effects. tissues –33. HoweThis study investigates the effects and possible underlying mechanisms of osthole activity on cell death and migration in both human GBM cells and in a glioma cell line selected for a high migration potential. Our study confirms previous reports that osthole induces cell death in human glioma cells, and indicates that osthole effectively inhibits FAK activation, MMP-13 expression, and cell migration.2.2.1.In order to investigate the growth inhibitory effects of osthole, U251 and HS683 human glioma cells were incubated with various concentrations of osthole  for 24 h, and the rate of growth inhibition was determined by MTT and SRB assay. We observed that osthole suppressed the growth of human glioma cells in a dose-dependent manner .2.2.Transwell assays were performed to investigate the effects of osthole on human glioma cell migration. Osthole-regulated glioma cell migration was examined using the transwell assay. As shown in 2.3.It has been reported that MMP-13 and FAK expression is involved in cancer cell migration. As shown in 2.4.We selected U251 and HS683 cell with high cell mobility, as described in Materials and Methods. This migration-prone subline (P10) had higher cell mobility and migrated more easily through the cell culture insert basement membrane matrix than the original U251 and HS683 cells  and dissolved in DMSO (dimethyl sulfoxide). The final concentration of DMSO in this study did not exceed 0.1% and had no effect on cell viability. Fetal bovine serum (FBS) and Dulbecco’s modified Eagle’s medium (DMEM) were purchased from Gibco BRL . Primary antibodies against phosphorylated FAK (Tyr397) were purchased from Cell Signaling and Neuroscience . Primary antibodies specific for MMP-13 or β-actin were purchased from Santa Cruz Biotechnology .4.2.2 flasks with DMEM. Both cell lines were cultured in medium supplemented with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin at 37 °C, incubated in a humidified atmosphere consisting of 5% CO2 and 95% air.U251 and HS683 cells originated from a human brain glioma. All cell lines were purchased from the American Type Culture Collection , and maintained in 75 cm4.3.4 per well for 24 h. Following treatment with osthole for another 24 h, medium was removed and cells were washed with PBS. MTT (0.5 mg/mL) was then added to each well and the mixture was incubated for 2 h at 37 °C. MTT reagent was then replaced with DMSO (100 μL per well) to dissolve formazan crystals. After the mixture was shaken at room temperature for 10 min, absorbance was determined at 550 nm using a microplate reader .Cell viability was determined using the 3--2,5-diphenyltetrazolium bromide (MTT) assay according to our previous study . U251 an4.4.4 per well for 24 h. After treatment with osthole for another 24 h, cells were fixed with 10% trichloroacetic acid and stained with SRB at 0.4% (w/v) in 1% acetic acid for 30 min. Unbound SRB was washed out with 1% acetic acid and SRB-bound cells were solubilized with 10 mM Trizma base. The absorbance was read at a wavelength of 515 nm using a microplate reader .The SRB assay is based on the measurement of cellular protein content according to our previous study . U251 an4.5.Whole cell lysis was performed as described in our previous report . Briefly4.6.in vitro migration assay was performed using Costar transwell inserts  in 24-well plates as described previously [4 cells in 200 mL serum-free media were placed in the upper chamber, and 500 mL of the same mixture containing osthole at various concentrations were placed in the lower chamber. Plates were incubated for 24 h at 37 °C in 5% CO2, and then cells were stained with 0.05% crystal violet and 2% methanol in PBS for 15 min. Non-migratory cells on the upper surface of the filters were removed by wiping with a cotton swab and then washed with PBS. The cell number was counted (5 fields per well) using a microscope at 100× magnification and expressed as the “fold of control”. Images of migratory cells were observed and acquired at 24 h with a digital camera and light microscope.The eviously . Approxi4.7.Procedure of wound-healing assay was performed as described in our previous report . Briefly4.8.2, 1 mM ZnCl2, 0.02% NaN3, and 1% Triton X-100) at 37 °C for 24 h. Gels were subsequently stained with Coomassie brilliant blue and destained in buffer containing 50% methanol and 10% acetic acid (v/v). The location of gelatinolytic activity was detected as clear bands.Samples of supernatant medium conditioned by cell culture under different experimental condition were centrifuged. Samples in equal volume were separated on a 10% SDS-polyacrylamide gel containing 0.1% gelatin. After electrophoresis, gels were washed with 2.5% Triton X-100 (in 50 mM Tris–HCl) for 30 min to remove SDS. Substrate digestion was formed by incubating the gel in developing buffer (50 mM Tris–Hcl containing 5 mM CaCl4.9.Subpopulations from glioma cells were selected according to their differential migration ability; the cell culture insert system was used as described earlier. After 24 h of migration, cells that penetrated through pores and migrated to the underside of the filters were trypsinized and harvested for a second-round selection. Original cells that did not pass through membrane pores were designated as P0. After 10 rounds of selection, the migration-prone subline was designated as P10.4.10.t-test. The difference was significant if p < 0.05.The values given are means ± S.E.M. The significance of difference between the experimental group and control group was assessed by the Student’s 5.We have shown that osthole effectively induces cell death in human glioma cells, and also that this agent significantly reduces FAK phosphorylation and MMP-13 expression in human glioma cells. Cell shape is involved with cell activity and may be considered a critical determinant of cell function . Recent


Eucalyptus camaldulensis and E. tereticornis are closely related species commonly cultivated for pulp wood in many tropical countries including India. Understanding the genetic structure and linkage disequilibrium (LD) existing in these species is essential for the improvement of industrially important traits. Our goal was to evaluate the use of simple sequence repeat (SSR) loci for species discrimination, population structure and LD analysis in these species. Investigations were carried out with the most common alleles in 93 accessions belonging to these two species using 62 SSR markers through cross amplification. The polymorphic information content (PIC) ranged from 0.44 to 0.93 and 0.36 to 0.93 in E. camaldulensis and E. tereticornis respectively. A clear delineation between the two species was evident based on the analysis of population structure and species-specific alleles. Significant genotypic LD was found in E. camaldulensis, wherein out of 135 significant pairs, 17 pairs showed r2≥0.1. Similarly, in E. tereticornis, out of 136 significant pairs, 18 pairs showed r2≥0.1. The extent of LD decayed rapidly showing the significance of association analyses in eucalypts with higher resolution markers. The availability of whole genome sequence for E. grandis and the synteny and co-linearity in the genome of eucalypts, will allow genome-wide genotyping using microsatellites or single nucleotide polymorphims. EucalyptusPopulusEucalyptus hybrids, LD was estimated with random amplified polymorphic DNA (RAPD) markers Pinus radiataVitis vinifera an out-crossing perennial species with high diversity, the utility of SSR markers were demonstrated for genome wide analysis Linkage disequilibrium (LD) estimation is currently in focus for plant breeding because of its application in association mapping of quantitative and adaptive traits in natural populations. Association mapping or association analysis is a method to study the relationship between phenotypic variation and genetic polymorphism Eucalyptus camaldulensis and E. tereticornis due to their wider adaptability to various types of edaphic and climatic conditions. In natural locations, these species occur in sympatry, particularly in Queensland region  and overlapping flowering period facilitates interspecific hybridizations E. camaldulensisEucalyptus species showed that LD decays within several hundreds of base pairs and suggesting the possibilities of much finer mapping resolution in LD based association mapping Eucalyptus using multiallelic SSR markers. Our specific objectives were to (1) discriminate the species and investigate population structure within selected germplasm of E. camaldulensis and E. tereticornis (2) determine the extent and genomic distribution of LD between pairs of SSR alleles to analyze the upcoming opportunities for association mapping in eucalypts.Eucalypts are one of the predominant tree species exploited for the paper pulp production. The tropical eucalypt plantations in countries like India are mainly occupied by E. grandis, E. nitens and E. tereticornis) were cross amplified in E. camaldulensis and E. tereticornis and 62 primers (56.9%) amplified one or two bands and others showed either no amplification or spurious pattern. A total of 1067 alleles were detected in 93 individuals amplified with 62 microsatellites. The summarized data on polymorphic information content (PIC), major allele frequency, observed and expected heterozygosity for the accessions of E. camaldulensis, E. tereticornis and its hybrids are given in E. camaldulensis and E. tereticornis showed high mean PIC value of 0.84. To explore genetic diversity among genotypes within the species, estimates of observed and expected heterozygosity and average number of alleles was also calculated. The observed heterozygosity varied from 0.44  to 0.95 , with an average of 0.87. Similarly, the expected heterozygosity values ranged from 0.36 to 0.92 with the average of 0.69 for all eucalypt accessions. The detailed data for each microsatellite loci is given in supporting information . Among the 62 SSR loci analysed 55 loci (89%) were polymorphic and 7 loci were monomorphic across all the three groups and 17 SSR loci were monomorphic between the two species. Analysis of 38 microsatellite loci (61%) for the presence of most common alleles with GDA software showed that 23, 14 and 38 SSR alleles were specific for E. tereticornis, E. camaldulensis and the landraces respectively  formed a group and other accessions of E. camaldulensis formed a separate group  and partitioning of the variation was highest within individuals (74%). Similar amount of variance was detected with K = 7 estimated from STRUCTURE, but with higher STF values between the populations  . However, estimation of LD was conducted by considering two species as individual group of populations.The genetic structure of the eucalypts collection was analyzed with the STRUCTURE program with 62 SSRs. Initially the two species were considered as individual populations and hence STRUCTURE was used not to determine the numbers of populations but to assign individual genotypes to the two populations. In K = 2, te group . We haveulations . AnalysiP<0.000)  and veryAB) between most common pairs of alleles among all loci using GDA software. In E. camaldulensis out of 135 significant pairs , 18 pairs showed r2 in the range of 0.05 to 0.09 and 17 pairs showed r2≥0.1. Similarly, in E. tereticornis out of 136 significant pairs, 18 pairs showed r2≥0.05 to 0.09 and 18 pairs showed r2≥0.1. In the E. camaldulensis accessions, pairwise r2 estimates among 62 loci varied from 0.00 to 0.133 with a mean of 0.09 and in E. tereticornis r2 varied from 0.00 to 0.62 with a mean of 0.012.The eucalypts species were considered separately for estimation of LD, since speciation increases linkage disequilibrium. Assuming the absence of Hardy-Weinberg equilibrium (HWE) in the selected population, the unphased genotypic data was used to estimate genotypic LD or composite LD coefficient  were considered for LD estimation. In E. tereticornis and E. camaldulensis accessions 46 and 15 allelic pairs were significant respectively  and showed interallelic r2>0.1. No allele pair with r2<0.1 was significant. The minimum and maximum interallelic r2 value for E. camaldulensis was 0.11 and 0.51 respectively with the mean of 0.19. Similarly, in E. tereticornis, the minimum and maximum interallelic r2 was 0.16 and 0.41 with the mean of 0.25.Haplotypic interallelic LD among all loci distributed over 11 linkage groups for the two species were estimated using reconstructed haplotypic data as implemented in MIDAS software by avoiding the assumption of HWE. The most frequent alleles represented by N/Y and Y/Y combinations  was = 0.17 and 0.14, and haplotypic r2 was = 0.39 and 0.31 for E. camaldulensis and E. tereticornis respectively. The r2 values above these threshold were probably caused by genetic linkage 2 estimates for genotypic data . To determine the relevance of these alleles (r2>0.1) for association analysis, identification of more number of SSR alleles with higher LD may be required. Between the species, accessions from E. tereticornis showed more number of allele pairs in LD than E. camaldulensis.The LD decay plots of the genotypic data are displayed in pic data  was beloE. camaldulensis and E. tereticornis. Analysis and characterization of microsatellites in Eucalyptus species have detected loci with highly variable number of alleles, heterozygosity levels and PIC values Symphyomyrtus. In E. camaldulensis, the PIC and heterozygosity of SSR loci ranged from 0.48 to 0.99 (average of 0.84) and 0.25 to 0.90, respectively E. camaldulensis, Butcher et al. The transferability of SSR loci among the species of eucalypts is amply represented E. tereticornis were reclassified under E. camaldulensis and many provenances show mixed morphology between the two species E. camaldulensis and E. tereticornis with 38 loci could differentiate both the species due to the presence of species-specific most common alleles. Although limited number of landrace accessions were used, the presence of SSR alleles belonging to both the species was obvious, suggesting that these SSR markers could be used to identify pure species and their hybrids.In eucalypts, the predominance of natural hybrids is well documented and landraces are considered as putative hybrids Populus nigraPseudotsuga menziesii var. menziesii, where the SNP marker based LD was very low (r2<0.05) Populus nigra decay of r2 with distance in CAD4 gene was observed at about 16 bp Eucalyptus globulus 20 wood quality candidate genes was analysed using SNP markers and LD was estimated to decay rapidly except in few genes where LD extended beyond 500 bp Populus nigra and P. balsamifera genome had rapid LD decay across the gene sequences 2 was well below 0.05 when estimated with SSR markers. In perennial fruit species like Prunus persica and Vitis vinifera, having a history of domestication and breeding showed a long range LD among the cultivated varieties Vitis vinifera L. subsp. silvestris LD decayed rapidly, with r2 values decreasing to 0.1 within 2.7/1.4 cM for genotypic/haplotypic data 2 was very low   , since t2, the interallelic LD estimated in E. camaldulensis and E. tereticornis was high (mean r2 = 0.19 and 0.22), indicating the possibilities of using SSRs for association analysis . Similarly, Inghelandt et al. The haplotype based interallelic LD, employed in human and animal populations for LD and association analysis Pinus radiata, LD was tested in full sib families created with limited number of pollen and seed parents , using 34 SSR markers and significant correlation was observed between trait and marker loci Most of the LD estimation studies in tree species are conducted on candidate genes The basis for linkage disequilibrium based association analysis in plant breeding is to ultimately develop markers tightly linked to trait loci or to identify exact causal loci for marker assisted selection E. camaldulensis and E. tereticornis, which has high implications in genetic improvement. It provides an understanding of how LD varies in the genome of outcrossing forest trees, hitherto available only with small genomic regions.In conclusion, the presence of species-specific alleles and population structure of the sample suggests that clear species delineation occur and hence separate management of these two species is highly essential in introduced countries to maintain species purity and progeny heterosis. The microsatellite information generated in this study has broadened our understanding on the linkage disequilibrium of the two important species, E. camaldulensis and E. tereticornis used in this study is available as supporting information , 8 landraces of India and 10 F1 putative interspecific hybrid trees, where the maternal parent is E. tereticornis (henceforth referred as F1 hybrids).The background of the accessions belonging to ormation . The gerE. grandis, ten primers from E. grandis and E. nitens mapped by Thamarus et al. E. tereticornis and 1 SSR (EMCRC 47) from Corymbia citriodora subsp. variegata mapped by Shepherd et al. Eucalyptus accessions. The PCR amplification was carried out in 10 µl volume containing 0.4 µM of each primer, 1 unit of Taq DNA polymerase, and 0.4 mM of each dNTPs, 1× buffer (with 200 ng BSA) and 10–20 ng of template DNA. The PCR amplification was carried out for 5 min at 94°C, 30 cycles of 1 min at 94°C, 60 or 30 sec at the primer specific annealing temperature, 2 min at 72°C, and 7 min at 72°C for final extension. Annealing temperatures varied from 48°C to 60°C, to amplify specific microsatellite markers. PCR products were size-separated using an 5% denaturing polyacrylamide gels of size 21 cm×50 cm  containing 7 M urea and 1× TBE buffer, and visualized by silver staining. The PCR products of the 93 accessions were run randomly for each primer and plate to avoid scoring errors among plates. Sixty two primers producing one or two clear bands were selected for further analysis  manually using the 50 bp size standard . Observed and expected heterozygosity, allele frequencies and polymorphic information content (PIC) values for each primer were calculated using the PowerMarker software STF estimation was conducted using the program Arlequin, v. 3.0 Presence of most common alleles in 40 individuals of AB) between pairs of common alleles at two loci were calculated. Calculations were based on Weir's method 2AB, as in Barnaud et al. P<0.05) of disequilibrium between all pairs of loci.The phases between alleles at two heterozygous SSR loci were unknown, hence composite linkage disequilibrium (LD) coefficients  and most frequent alleles (Y). In this study N/Y and Y/Y were only considered for LD estimation as the rare alleles may skew the linkage disequilibrium estimates The interallelic LD computation was performed using the software MIDAS  2 values against the distance (cM) between markers on the chromosome. Map positions of all mapped SSRs were based on the consensus map developed by Brondani et al. E. grandis X E. urophylla and Thamarus et al. E. globulus. The consensus map was selected in this study because a high degree of co-linearity and synteny between the genetic maps of various eucalypts species was demonstrated including E. tereticornisth percentile of that distribution was taken as a population specific critical value of r2, beyond which LD was likely to be caused by genetic linkage. The overall decay of LD (r2) with physical distance (cM) among the SSR loci was evaluated by nonlinear regression (NLR) 2 with our data.The decay of LD over genetic distance was investigated by plotting pairwise rTable S1SSR loci used in this study showing information on linkage group, annealing temperature, amplification range, major allele frequency, polymorphic information content, observed and expected heterozygosity.(DOC)Click here for additional data file.Table S2Details on eucalypts accessions used in this study.(DOC)Click here for additional data file.


Similar to other developed countries, only 3% of the total population in Hong Kong donate blood (Hong Kong Red Cross Blood Transfusion Service 2003). More than 20% of annual donations come from youngsters aged 18-25. However, this category of donors has decreased gradually from 24.6% in 2004 to 22.9% in 2008. This study aims to examine the characteristics and intention of young blood donors versus nondonors in Hong Kong; and to explore the factors that may influence Hong Kong young people’s donation behavior.This is a cross-sectional study using questionnaire to solicit information from young people including both blood donors and non-donors.It showed that more non-donors were underweight (26%) than blood donors (16.9%). Blood donors demonstrated to have more knowledge on the usage of donated blood (87.2%). Nearly half of youngster admitted that they made use of donation as a means for blood testing (53.1%) or free physical check-up (47.3%).Recruitment strategies should focus on the enhancement of health education programs related to blood and blood donation for young people to increase their awareness of blood and alleviate their misconceptions about blood donation. Blood is a vital healthcare resource; thus, a reliable and sufficient blood supply is of paramount importance in healthcare setting and blood donors are always the primary focus of blood transfusion service. Similar to many countries over the world, only 3% of the total population in Hong Kong donate blood. More thaAccording to the Hong Kong Government projection, 27% of the population will be over 65 years of age by 2033.[Besides, a total of 451,781 patients aged 70 and above were admitted into the hospitals under Hospital Authority in 2008 and the increase was 7.1% over the preceding year.[The objectives of this paper are (1) to examine the characteristics and intention of young blood donors versus non-donors in Hong Kong; (2) to explore the factors that influencing Hong Kong young people’s donation behaviors.As information about Hong Kong young people’s attitudes and behavior toward blood donation is limited, a cross-sectional survey was performed to profile the current situation. A structured self-administered questionnaire was used to solicit information from young people.A newly refined and structured questionnaire was developed. The “theory of planned behavior” was adopYoung Chinese people aged 18 to 25, including both blood donors and non-donors, were the targeted population of this study. The donors were recruited from multiple blood donation centers, and the non-donors were recruited from schools/ institutes including those who accompanied their friends to the centers but did not donate blood .A total of 3,316 young people were recruited in August and September 2008. The response rate was 94.7%. Among the respondents, 2,523 (76.1%) were blood donors (BD) and 785 (23.7%) were non-donors.P<0.0001. They showed to have a higher BMI  than nondonors (71.1% with P<0.0001). Non-donors were significantly underweight (28.9%) and 22.3% of them were born outside Hong Kong. They also resided in Hong Kong for only a short period of less than 10 years (12.0%) [Blood donors in this study were statistically significant heavier (57.9 kg) than nondonors (55.3 kg) with  (12.0%) .Among the blood donors, more than half of them had two to five donations (54.1%); however, only 3% of them donated blood within an interval of one year. Blood donors used to donate blood at static donor centers (73.4%). Majority of blood donors did not encounter any adverse reaction during donation (73.7%). About one tenth of blood donors experienced mild reactions including dizziness (9.8%), bruise/hematoma (8.9%), or feeling fatigue (3.3%) in their blood donations. A small proportion of donors came across severe reaction that had led to subsequent injuries such as fatally faint (0.5%). Majority of blood donors satisfied with the BTS’ services . However, some of them disagreed with the long waiting time for donating blood (18.9%).vs ND: 16.1%, P<0.0001), the usage of donated blood (BD: 87.2% vs ND: 82.9%) and realized that blood was needed to save life (BD: 94.8% vs ND: 90.9%). Blood donors perceived they had possessed better health (BD: 96.6% vs ND: 90.6%) and less likely practised in high-risk behaviors (BD: 95.7% vs ND: 92.7%). Blood donors more actively participated in other kind of donation than non-donors. Many of them had registered as organ donors . Unfortunately, a small proportion of respondents revealed that they had donated blood even though they engaged in high-risk behavior such as having sex with strangers (1.3%) and more than one sexual partner (1.0%).Blood donors demonstrated better knowledge on blood and blood donation related issues than the non-donors. Most of blood donors knew their blood type  and blood testing of HIV / Hepatitis B (22.9%). They also agreed more with the barriers impacting their blood donation behavior such as afraid of pain (45.8%), afraid of needle (34.3%), afraid of sight of blood (20.3%), too thin (22.5%), afraid of being fainted when donating blood (15.5%), having no time (21.6%), afraid of affecting their health (14%).P<0.0001). However, some blood donors agreed with the poor service provided by BTS which included the long waiting time for blood donation  and the inconvenient operation hours of donation centers . Non-donors shared similar findings as they also considered it, the long waiting time for donation (15.5%) and the inconvenience of BTS’ operating hours (19.4%). The results also revealed a clear negative effect that respondents disagree with being informed by BTS of not suitable for blood donation (BD: 9.4% and ND: 15% with P<0.0001).A statistically significant high percentage of blood donors agreed with blood transfusion service’s (BTS) good reputation (91.8%), professional knowledge (95.6%) and excellent service (94.2%) than those of non-donors  and blood donation was beneficial to their health (BD: 61.2% vs ND: 51.9%). However, respondents also indicated that they had not been requested by other people for blood donation (BD: 53.4% vs ND: 38.1%). Blood donors considered their blood type was important (BD: 37.9% vs ND: 25.6%). They also agreed with their friends’ expectation for blood donation (BD: 32.8%) than non-donors .Blood donors had more positive attitude toward blood donation than non-donors. Blood donors agreed that donating blood was very rewarding as people will reciprocate to them whenever they were in need (BD: 81.6% vs ND:19.3%). In contrast, non-donors disagreed that they had a duty to donate blood (72.7%) and blood donation was the moral obligation .Blood donors considered blood donation was a kind of social responsibility (83.3%) and moral obligation (64.9%). Besides, more blood donors had family members donating blood (BD: 29.4% vs ND: 35%), “afraid of needles” (BD: 18% vs ND: 34.3%), “being too thin” (BD: 6.7% vs ND: 22.5%), “having no time” (BD: 11.1% vs ND: 21.6%), “being afraid of the sight of blood” (BD: 9.7% vs ND: 20.3%), “would faint if donate blood” (BD: 4.9% vs ND: 15.5%) and “donation may affect their health” (BD: 3.9% vs ND: 14%) [Blood donors show significantly stronger control power than non-donors. Non-donors concern more about the barriers than blood donors which include “they are not sure about the donation procedures” (BD: 7.3% ND: 14%) .vs ND: 42.5%), having the intention to donate (BD: 76.3% vs ND: 48.5%) or having intention to try (BD: 74.7% vs ND: 61%). More blood donors also agreed with being influenced by BTS information to donate blood (BD: 47.7% vs ND: 33.5%) .Blood donors demonstrated significantly greater intention to give blood. The majority of blood donors indicated that they had decided to donate (BD: 83.6% Most of the blood donors are heavier in weight (59.6%) than the non-donors. Besides, a significant underweight condition among young people has been identified in this study. This will eventually affect the eligibility of young people for blood donation in future. The study results also show that non-donors tend to be much lighter and more than one forth of them are underweight (28.9%) whereas the percentage of underweight non-donors is double than that of donors in 21 to 25 years range for both males and females. Since body weight is one of the crucial criteria for blood donation, youngsters of low body weight will be excluded for donation of blood.Blood donors prefer to make their donation at static donation centers 73.4%) than at mobile collection teams (25.5%) and blood donation vehicles (1.1%). However, non-donors consider donating blood is inconvenient as it takes a long waiting time (15.5%). They also find the location of donation centers and their operation hours unsuitable (28.8 and 19.4% respectively). Findings from a previous Hong Kong study indicated that there was a problem dealing with those blood donors who preferred the door-to-door service and were only willing to donate blood during the visits of the mobile collection teams to their workplace or near their home.[3.4% thanIn Hong Kong, a large number of potential donors were lost due to temporary pre-donation deferral every year. There were 32,014 potential donors deferred in 2009. Among them, 50.6% were rejected because of low hemoglobin level, 11.0% were under medical care and 9.7% were not feeling well at the time before donation. The results of this study also indicate that a certain percentage of young people admit that they have been informed by BTS for not suitable to give blood (BD: 9.4% and ND: 15%). Among these young people, particularly the non-donors, temporary deferral may psychologically lead to an excuse for not donating in future and inteAlthough a significant number of young people understand the usefulness of blood donation  and also know that blood is needed for saving life , it remains unknown why the non-donors of this category lacks of willingness to donate. A lack of relevant information about blood or blood donation is reported to be a cause of keeping many young people away from blood donation, and it is commonly used by the non-donors as an excuse of not giving blood.Some non-donors indicate that, even though their family members had received blood transfusion in the past (18.6%), they never ever donate. The above data let us to infer that strategies have to be formulated to encourage this group of individuals to reciprocate those in need.On the other hand, blood donors point out that they expect people to reciprocate them whenever they are in need (BD 81.6% and ND 74.1%). This finding implies that blood donation behavior may not be driven out of pure altruism – the behavior that only concerned the welfare of others regardless of personal benefits. Blood reThe majority of blood donors in the current study perceive they are able to overcome obstacles and barriers for blood donation without great difficulty. In consistent with the previous researches,–14 non-dYoung people in fact do not oppose the idea of donating blood to save lives. Yet motivating people to donate blood is undoubtedly a difficult task. Provision of effective health education programs from BTS to the general public is one of the crucial factors in recruiting and retaining blood donors. BTS can convey updated knowledge to the young people to increase the awareness of blood donation and alleviate the misconceptions about blood donation among the general public.


Aspergillus fumigatus conidia. On average, 25% of macrophage-associated conidia were phagocytosed after one hour. Sixteen percents of these cell-associated conidia were killed after 4 h and conidial germination was inhibited in more than 95% of the conidia. A. fumigatus conidia were shown to be cytotoxic to the macrophages. Intracellularly germinating conidia were located free in the cytoplasm of necrotic cells, as shown using transmission electron microscopy. These results suggest that avian respiratory macrophages may prevent early establishment of infection, unless the number of A. fumigatus conidia exceeds the macrophage killing capacity, leading to intracellular germination and colonization of the respiratory tract.Although aspergillosis is one of the most common diseases in captive birds, the pathogenesis of avian aspergillosis is poorly known. We studied the role of avian respiratory macrophages as a first line of defense against avian aspergillosis. The phagocytic and killing capacities of avian respiratory macrophages were evaluated using pigeon respiratory macrophages that were inoculated with  Aspergillus species are a major cause of morbidity and mortality among captive birds [A. fumigatus is the predominant species found in infected birds [Fungal infections caused by ve birds . The filed birds . This fued birds .A. fumigatus occasionally seem to be able to germinate within alveolar macrophages [The primary route of infection in birds is inhalation of these airborne conidia, resulting in lower respiratory tract colonization . In mammrophages ,12.In comparison to the mammalian respiratory system, a paucity of free respiratory macrophages exists in the avian respiratory system -16. HoweDue to predominant colonization of the avian lower respiratory tract, avian respiratory macrophages can fulfill an important defensive role during the initial colonization events. Therefore, the aim of this study was to assess the role of the avian respiratory macrophage in avian aspergillosis.A. fumigatus isolate, K24, used in the in vitro studies, was obtained from a racing pigeon, which died from pulmonary aspergillosis [A. fumigatus conidia. The conidia were washed three times in 0.01% Tween 20 in RPMI and the suspension was adjusted to a concentration of 106A. fumigatus conidia/mL in RPMI 1640 with 1% glutamine and 1% pyruvate by haemocytometer count.The gillosis . Five-daPigeon respiratory macrophages were collected according to a method as previously described , and maiA. fumigatus conidia, pigeon respiratory macrophages were counted and seeded into a 96-well plate at 105 macrophages per well. After 2 h of incubation at 37°C and 5% CO2, the wells were rinsed to remove non adherent cells. The pigeon respiratory macrophages were exposed to 0.2 mL of 106 conidia/mL in RPMI 1640 (+ 1% glutamine and 1% pyruvate). The plates were centrifuged at 125 × g for 10 min to synchronize conidial exposure to the cells. Subsequently, the cells were allowed to ingest the conidia for 1 h at 37°C, 5% CO2. Medium containing non adherent, non phagocytosed conidia was removed, and wells were rinsed three times using HBSS with Ca2+ - Mg2+ at 37°C and fresh medium was added (time point 0). The macrophages were then further incubated for 4 h before cell associated conidia were harvested (time point 4). At time points 0 or 4, plates were frozen at -70°C and rapidly thawed at 37°C to lyse the cells and harvest the conidia. Cellular lysis was confirmed by light microscopy. Serial dilutions were performed in sterile medium and immediately plated on Sabouraud dextrose agar. Plates were incubated at 37°C, and colonies were counted after 24 h of incubation. Negative control wells did not contain macrophages but were otherwise treated likewise. The percentage killed conidia was calculated by dividing the total number of conidia recovered at 4 h by the total number of conidia recovered at 0 h and multiplied by 100. All tests were performed in triplicate.To assess the killing capacity of avian respiratory macrophages against A. fumigatus conidia, pigeon respiratory macrophages were seeded in 24-well plates and exposed to 1 mL of 105 conidia/mL. The plates were centrifuged at 125 × g for 10 min to synchronize conidial exposure to the cells. After phagocytosis for 1 h, medium containing non adherent, non phagocytosed conidia was removed and the wells were rinsed three times using HBSS with Ca2+ - Mg2+ at 37°C. To discriminate between ingested and adherent conidia, the cells were further incubated with Calcofluor White M2R , which stains the fungal cell wall of extracellular, but not that of intracellular conidia. For quantification of phagocytosis, the cells were then washed and fixed with 4% (v/v) para-formaldehyde for 10 min at room temperature. Per well, 100 conidia were analyzed for intra- vs. extracellular location using fluorescence microscopy. All tests were performed in triplicate.To assess the percentage phagocytosed 5 conidia/mL. The plates were centrifuged at 125 × g for 10 min to synchronize conidial exposure to the cells. After phagocytosis for 1 h, medium containing non adherent, non phagocytised conidia was removed and the wells were rinsed three times using HBSS with Ca2+ - Mg2+ at 37°C. To discriminate between ingested and adherent conidia, the cells were further incubated with Calcofluor White M2R. The cells were further incubated for 6 h and then fixed with 4% (v/v) para-formaldehyde for 10 min at room temperature. Per well, 100 conidia were analyzed for germination activity using fluorescence microscopy. The tests were performed in triplicate. To observe the intracellular trafficking and intracellular germination of A. fumigatus conidia electron microscopically, pigeon respiratory macrophages were seeded in 24-well plates at 5 × 105 / mL and incubated for 2 h at 37°C and 5% CO2. After the incubation period, the wells were rinsed to remove non adherent cells and subsequently infected with 5 × 105A. fumigatus conidia. The wells were then centrifuged at 125 × g for 10 min at 37°C to synchronize conidial exposure to the cells. At time points 0, ½, 1, 2, 4 and 8 h, the cell medium was removed and the infected cells were fixated in 4% formaldehyde containing 1% CaCl2 (w/v) in 0.121 M Na-cacodylate adjusted to pH 7 for 24 h. Then the cells were washed in Na-cacodylate buffer and preserved in the Na-cacodylate buffer. The samples were then divided into smaller portions (1 mm2) with a razor blade, dehydrated in graded ethanol solutions, and embedded in LX112 resin . Semithin sections (2 μm) were stained with toluidine blue to select the most appropriate regions for ultrathin sectioning. Ultrathin sections of 90 nm were cut with a diamond knife on a Reichert Jung Ultracut microtome , mounted on formvar-coated copper grids, and stained with uranyl acetate and lead citrate. The samples were examined under a Jeol EXII transmission electron microscope  at 80 kV.To quantify intracellular germination, pigeon respiratory macrophages were seeded in 24-well plates and exposed to 1 mL of 10A. fumigatus on avian respiratory macrophages, pigeon respiratory macrophages were seeded in 24-well plates. From each of 5 pigeons, one well with pigeon respiratory macrophages was exposed to 1 mL of 105 conidia/mL and one well was left untreated. The plates were then centrifuged at 125 × g for 10 min. After 6 h of incubation, the wells were rinsed and exposed to ice cold ethidium monoazide (EMA) solution (50 μg/mL) for 20 min in the dark. The wells were then exposed to an incandescent lamp (45W) for 10 min. Next, the cells were rinsed and fixed with 4% (v/v) para-formaldehyde for 10 min at room temperature. After washing, the cells were stained with Hoechst (10 μg/mL) for 15 min at room temperature. Per well, 100 macrophages were analyzed for viability using fluorescence microscopy.To determine the cytotoxic effect of 5/mL) were seeded into a chamber slide system in sterile RPMI 1640 with glutamine medium supplemented with 10% foetal bovine serum and 1% penicillin/streptomycin and incubated overnight. After 2 h, the cells were rinsed three times with HBSS at 37°C. Cells were inoculated with 1 mL of 105 conidia/mL in RPMI 1640  and the plates were centrifuged at 125 × g for 10 min. The cells were allowed to ingest the conidia for 1 h. Medium containing non adherent, non phagocytosed conidia was removed, and wells were rinsed three times using HBSS with Ca2+-Mg2+. To discriminate between ingested and adherent conidia, the macrophages were further incubated with Calcofluor white M2R with a final concentration of 25 μM. After 30 min, the cells were washed and resuspended in RPMI 1640 . The chamber slide dishes were placed on an Olympus IX81 fluorescence microscope  connected to a Cell*M imaging system (Olympus), and equipped with a Hamamatsu B/W Orca camera , a Märzhaüser automated microscope table , and a CellCubator climate chamber (Olympus). Bright-field images of 12 cells were collected every 15 min for 15 h.To verify intracellular germination and cellular lysis, pigeon respiratory macrophages  (P < 0.01) and significantly less apoptotic  cells (P < 0.05) than control cells. In infected wells, on average 22.8% +/- 4.7 of the cells were necrotic and 5.4% +/- 2.5 were apoptotic whereas 9.2% +/- 3.7 were necrotic and 11.2% +/- 4.9 were apoptotic in the non-infected wells  P < 0.0 and signIntracellular germination in the cytoplasm of pigeon respiratory macrophages was observed in a small portion of these cells. Six hours after ingestion, the fluorescence assay demonstrated that intracellular germination of conidia occurred in 2% +/- 1 of the pigeon respiratory macrophages Figure  while soA. fumigatus conidia. The pigeon respiratory macrophages demonstrated a limited capacity of killing conidia in the first 5 hours post infection, compared to mammals [A. fumigatus conidia, rendering the avian macrophage a highly efficient first line defense against A. fumigatus. Inhibition of germination was also observed in alveolar macrophages derived from mice, rabbits and humans [A. fumigatus infections in birds [This is the first study that reports the phagocytic and killing capacity of avian respiratory macrophages against  mammals ,8-10. Hod humans . Even thin birds .A. fumigatus and is associated with cytotoxicity and cell death in murine macrophages [A. fumigatus conidia may overwhelm this defense mechanism, which perhaps may lead to colonization of the respiratory tract through intracellular germination of a limited number of conidia that caused macrophage cell death [A small proportion of internalised conidia was still capable of germinating inside the avian macrophages. Intracellular germination was associated with degeneration, oncosis and necrosis of the macrophages. The damage to the macrophages may be elicited by gliotoxin. Gliotoxin is a toxin produced by rophages . Taking ll death ,22,24.A. fumigatus conidia was either killed or their germination inhibited by avian respiratory macrophages. However large amounts of A. fumigatus conidia in the respiratory tract can result in intracellular germination and lysis of the phagocytic cells, which may contribute to colonization of the respiratory tract.In conclusion, this study demonstrates that the majority of The authors declare that they have no competing interests.LVW, AM and FP participated in the design of the study. LVW, KDH, HF and SJL performed the experiments and LVW, RD, KDH, AM and FP analysed the data. LVW, AM, FP, RD and FH wrote the manuscript. All authors read and approved the final manuscript.


The suspicion of a hyperparatiroidism is mostly guided by the finding of an increase in serum calcium levels by routine measurements. Primary hyperparathyroidism is a common disease occurring in 0.2 to 0.5% of the population. The incidence in the United States is approximately 100000 new cases per year and increases with age affecting up to 2% of elderly people [From January 1995 to December 2009, 172 patients underwent operations for Hyperparathyroidism, 130 of these were Primary Hyperparathyroidism at our Department of General Surgery. Patients were divided into two groups: patients of ≤ 69 years old (Group A) and patients of ≥ 70 years old (Group B). The following variables were studied: demographic characteristics, co-morbidities, preoperative symptoms, laboratory values, operative procedures, postoperative complications and anatomo -pathological findings.Group A: 110 patients operated 25 were male, 85 were female with a M:F ratio of 0.3:1. Mean age at admission was 52.4 (SD±12.9). We reported a morbidity rate of 5.4% (6 patients) and a mortality rate of 0%. Group B: 20 patients operated 6 were male, 16 were female with a M:F ratio of 0.25:1. Mean age at admission was 74.2 (SD±3.7). We reported a morbidity rate of 5% (1 patient) and mortality rate 0%.Elderly patients with Hyperparathyroidism present a variety of symptoms that are often different from those seen in younger patients. They are more likely to manifest fatigue and psychiatric symptoms that are difficult to distinguish from those due to their age, therefore in the majority of cases the suspicion of hyperparathyroidism is guided by the finding of an increase in serum calcium levels on a routine measurement. If serum calcium level is high or if hypercalcaemia is discovered, measurement of PTH confirms the diagnosis ,3. Cervi


Acute kidney injury (AKI) is a common complication of critical illness and sepsis . Dosing 0-24/MIC ≥400 (Cmin >10 mg/l), gentamicin target was Cmax/MIC of 8 to 10. Maintenance doses were adjusted according to drug level simulation using a pharmacokinetic programme.Seventeen septic patients with AKI treated with vancomycin and seven patients with gentamicin on CVVH were included. In the vancomycin group, patients received the first dose of 1.0 g intravenously followed by 1.0 g/12 hours if not adjusted. In the gentamicin group, patients received a loading dose of 240 mg followed by a maintenance dose every 24 hours. The vancomycin maintenance dose was optimized to achieve AUC0-24/MIC ≥400 ratio was achieved in 67% of patients on the first day. The median gentamicin Cltot was 0.68 and 0.793 ml/minute/kg on the first and second day of the study. CRRT clearance accounted for about 50% of gentamicin Cltot found in a population without renal impairment (0.73 ml/minute/kg). The target Cmax/MIC ratio was achieved in 78% of patients after the first dose.The median vancomycin total clearance (Cltot) was 0.89 and 0.55 ml/minute/kg on the first and second day of the study. CRRT clearance accounted for about 50 to 60% of vancomycin Cltot found in a population with normal renal function (0.97 ml/minute/kg). Vancomycin serum concentrations after the first dose were below the required target of 10 mg/l as early as 6 hours in 10 patients. AUCCVVH at a filtration rate of 45 ml/kg/hour leads to high removal of both antibiotics. Due to rapid change in patient's clinical status it was impossible to predict a fixed dosage regimen. We recommend administration of unreduced loading dose and: blood sampling as early as 6 hours after first vancomycin dose; blood sampling 30 to 60 minutes after gentamicin administration and before the next dose; and the maintenance dose should be based on drug-level monitoring.


To evaluate prognostic factors that may influence local control (LC) of T1N0 glottic cancer treated by primary radiotherapy (RT) with 6 MV photons.We retrospectively reviewed the medical records of 433 consecutive patients with T1N0 glottic cancer treated between 1983 and 2005 by RT in our institution. All patients were treated with 6 MV photons. One hundred and seventy seven (41%) patients received 52.5 Gy in 23 fractions with 2.5 Gy/fraction, and 256 (59%) patients received 66 Gy in 33 fractions with 2 Gy/fraction.p = 0.035); involvement of anterior commissure ; fraction size of 2.0 Gy  and tumor biologically effective dose (BED) < 65 Gy15 .The median follow-up time was 10.5 years. The 10-year LC rates were 91% and 87% for T1a and T1b respectively. Multivariate analysis showed LC rate was adversely affected by poorly differentiated histology (Hazard Ratio [HR]: 7.5, The negative impact of anterior commissure involvement could be overcome by delivering a higher tumor BED through using fraction size of > 2.0 Gy. We recommend that fraction size > 2.0 Gy should be utilized, for radiation schedules with five daily fractions each week. Laryngeal cancer is the third most common head and neck (H&N) cancer in Hong Kong. The age-standardized incidence rate was 2.3 per 100,000   .Since the treatment field size for T1N0 GC is small, it permits slight hypofractionated schedule without causing excessive acute radiation toxicity. Shortened OTT overcomes the accelerated repopulation of tumor clonogens.This also supports the current contemporary practice of fraction dose size > 2.0 Gy (i.e. 2.25 Gy) for treatment of T1N0 GC by other centers ,21,24,37To the best of our knowledge, our report is the largest study on RT outcomes in T1N0 GC primarily treated with 6 MV photons. As the treatment of choice for early GC in our institution or Hong Kong at large has been and in the near future will still be RT alone , this re15). In order to achieve this tumor BED level in conventional schedule of five daily fractionation each week, we recommend that fraction size > 2.0 Gy should be utilized. In fact, modest hypofractionation is safe and effective for T1N0 GC in terms of both LC and morbidity. Having a shorter OTT is more convenient for patients and is also more cost- effective for RT facility implication.We suspect that the issue of 'cold spot' is more apparent at the AC region, especially when treated with 6MV photons. Certainly, further dosimetric evaluation is needed to validate this suspicion. While involvement of AC is an adverse prognostic factor, we have shown that its negative impact can be overcome by delivering a higher tumor BED (≧ 65 GyNevertheless, the results need to be interpreted with caution, because the current report was a retrospective, single institution study and therefore subjected to biases. For example, we did not have volume measurements on tumor, which has been shown in other reports as one of the important prognostic factors in LC ,19,25. IOur data concur with other published result about the efficacy of RT with 6 MV photons for T1N0 GC. While involvement of AC is associated with poor LC rate, its negative impact could be overcome by delivering a higher tumor BED through using fraction size of >2.0 Gy. We recommend that fraction size > 2.0 Gy should be utilized, for radiation schedules with five daily fractions each week.The authors declare that they have no competing interests.CCT participated in the study's design and coordination, performed acquisition of data and drafted the manuscript. KHA and FYC participated in data analysis and revised the manuscript. RKCN and SMC participated in study's design and revised the manuscript. JSKA, YTF and SCKL revised manuscript critically for important intellectual content. All authors read and approved the final manuscript.