Laboratory-based HIV tests for recent infection (TRIs), which primarily measure a specific serological biomarker(s) that distinguishes recent from long-term HIV infection, have facilitated the estimation of population-based incidence. Dried blood spots (DBS) on filter paper are an attractive sample source for HIV surveillance, given the simplified and cost-effective methods of specimen collection, storage, and shipment. Here, we evaluated the use of DBS in conjunction with an in-house multiplex TRI, the HIV-1-specific Bio-Plex assay, which measures direct antibody binding and avidity to multiple HIV-1 analytes. The assay performance was comparable between matched plasma and DBS samples from HIV-1 infected individuals obtained from diverse sources. The coefficients of variation, comparing the median antibody reactivity for each analyte between plasma and DBS, ranged from 2.78% to 9.40% and the correlation coefficients between the two sample types ranged from 0.89 to 0.97, depending on the analyte. The correlation in antibody reactivity between laboratory and site-prepared DBS for each analyte ranged from 0.87 to 0.98 and from 0.90 to 0.97 between site-prepared DBS and plasma. The correlation in assay measures between plasma and DBS indicate that the sample types can be used interchangeably with the Bio-Plex format, without negatively impacting the misclassification rate of the assay. Since the first AIDS cases were identified more than 30 years ago et al. described the development of a detuned or less-sensitive serologic assay for distinguishing recent from long-term HIV-1 infection, which allowed for incidence estimation from cross-sectional patient samples. Several serology-based laboratory tests for recent infection (TRI) have been developed that measure a specific biomarker, primarily HIV-1-specific antibody In 1998, a ground-breaking study by Janssen Given that the majority of current TRI approaches are serology-based, the most commonly used sample type for incidence testing is plasma and/or serum. Although HIV-1-specific antibodies are stable in plasma and serum, sample collection is somewhat limited due to processing and storage requirements. Dried blood spots (DBS) have been used extensively for HIV testing and surveillance, as they can be prepared from a finger stick, do not require centrifugation, and can be shipped at room temperature. HIV biomarkers remain stable on the DBS filter paper; with minimal risk of infectivity once the sample is fully dried To improve upon the accuracy of TRIs for estimating incidence, it has been demonstrated that multiassay algorithms (MAAs), combining one or more TRIs with clinical data, yield improved incidence estimates and reduced false-recent rates as compared to each individual test Longitudinal seroconversion panels were obtained through the Seroconversion Incidence Panel Project (SIPP) in collaboration with SeraCare Life Sciences, Inc. . The objective of the study was to identify recent HIV-1 seroconverters, as defined as having acquired HIV within the past 90 days. Donors were enrolled at plasma donation centers and HIV testing and/or clinical sites within the United States. Study participants were at least 18 years of age and met either of the following criteria: (1) detectable viral RNA with a negative or indeterminate antibody test result or (2) positive HIV antibody test with a documented negative test result within the past 90 days. Based on these criteria, 20 HIV-1 seroconverters were identified and enrolled for follow-up blood collection. Whole blood collection in EDTA vacutainers was initiated within 120 days from the date of the last negative or indeterminate HIV antibody test result. Blood samples were collected monthly for the first six months, followed by every three months for up to 15 months. Follow-up blood collections were continued after 15 months for most study participants . Twelve of the study participants had documented antiretroviral (ARV) use during the sample collection period. Cross-sectional samples from HIV-1 infected individuals with chronic or long-term infection were also collected. The last 2–3 samples collected from each of the seroconverters were included in the current study, along with 6 of the cross-sectional, long-term specimens (49 specimens/26 participants).Cross-sectional samples from HIV-1 infected men who have sex with men (MSM) were obtained through CDC's National Behavioral and Surveillance (NHBS) system, as part of the MSM3 cycle. HIV-1 infected MSM were identified through routine testing in five US cities , as described in detail Lastly, cross-sectional specimens were obtained from HIV-1 infected donors from Cameroonian blood banks, as described elsewhere For the SIPP and NHBS cohorts, whole blood was shipped to the CDC at ambient temperature, within 48 hours of collection. DBS were made from whole blood by aliquoting 50 µl blood per spot on Whatman 903 Protein Saver cards . Each card was dried overnight at room temperature and packaged individually in Bitran bags  with silica gel desiccant bags and humidity indicating cards . The DBS were stored at -20°C until ready for use. Matched plasma samples were prepared from the whole blood and stored at −80°C prior to testing. In addition to the CDC-prepared specimens, DBS were also prepared at the time of collection at one of the NHBS testing sites. Of the 51 NHBS participants included in this study, 14 had matched CDC and site-prepared DBS.Sample collection and storage for the Cameroonian samples have been described previously A standard paper punch was used to obtain a 6.3 mm (diameter) blood spot from the DBS cards. Each blood spot was transferred to one well in a flat-bottom 96-well plate with 250 µl pre-incubation buffer , and 0.8% polyvinylpyrrolidone (Sigma-Aldrich)) The HIV-1-specific Bio-Plex assay was performed on the matched plasma and DBS samples, as described To establish a baseline level of variation associated with the assay platform, intra-assay variation was assessed by calculating the coefficient of variation (CV) for the MFI values of each sample replicate. CVs were also calculated for the incident/prevalent standards included in each run. To evaluate variation associated with sample type, the CV comparing HIV-specific IgG antibody reactivity of plasma versus DBS was calculated for each analyte. The relationship between antibody reactivity of the plasma and DBS was also determined using Spearman's rank correlation coefficient.To determine whether sample type affects the ability of the Bio-Plex assay to discriminate between recent and long-term HIV infection, cutoff values were determined for each analyte. In the absence of finalized cutoff values for the overall assay, the range of reactivity for known recent and long-term samples was determined using an HIV-1 Incidence/Prevalence Performance Panel (SeraCare Life Sciences). Given the minimal to no overlap between the reactivity of the recent and long-term panel members, the top end of the 95% confidence interval for the reactivity of the recent samples was selected as a cutoff for the purposes of the current analyses. For the study specimens, recent infection was defined as any value below the analyte-specific cutoff.r) between assay values for plasma and DBS ranged from 0.89 to 0.97 for all cohorts. When analyzed separately, the avidity measures exhibited highly reproducible results for all cohorts (r≥0.9), while the correlation in normalized values was slightly more variable between cohorts (r≥0.8).Intra-assay reproducibility was relatively high for both plasma and DBS samples for all analytes . The medr values ranging from 0.87 to 0.98 for all analytes (r = 0.90 to 0.97). A comparison of the normalized MFI values and avidity index for the CDC versus site-prepared specimens is shown in The correlation in assay reactivity between CDC and site-prepared DBS was highly reproducible, with analytes . SimilarCutoff values for each analyte, chosen based on the assay reactivity of the performance panel, are listed in In previous studies, we described the development and performance characteristics of an in-house HIV-1-specific Bio-Plex assay for determination of recent HIV-1 infection with plasma specimens Given that most assays are subject to some degree of innate variation, it is important to characterize the baseline intra- and inter-assay variation prior to assessing potential differences in sample types. Specimen replicates yielded highly reproducible assay measures, irrespective of sample type. These findings were consistent with a previous study demonstrating performance of our in-house HIV-1 Bio-Plex assay versus a prototype kit, using plasma specimens r values of 0.99 or 1. This may be the result of well-controlled sample and data collection, along with the fact that the specimens included in the current study were collected from a single site and freeze-thaw cycles were minimal for the plasma specimens. The NHBS cohort also exhibited high correlation between sample types; however, a lower r value of 0.79 for gp160-n was skewed by a single outlier. Unfortunately, limited sample volume for this particular study participant prevented repeat testing. For this particular cohort, site-prepared DBS were also available for some of the study participants. We did not observe any differences in reactivity between the site and CDC-prepared samples, which demonstrates that samples can be collected at field sites, shipped, and stored for later testing. The Cameroon specimens were the oldest of the three cohorts, as they were collected and stored 10–15 years ago and the DBS had been freeze-thawed multiple times for previous testing. Additionally, the Cameroon cohort exhibited a large degree of HIV-1 subtype diversity. Despite the age of the specimens and subtype diversity, the correlation between sample types was high for all analytes. It is important to note that any differences in reactivity between sample types might be exaggerated given the small sample numbers included in current study.Specimens from three separate cohorts were evaluated in this study to assess potential inconsistencies due to site-specific sample collection or storage methods. Not surprisingly, the avidity measures were the most consistent between the cohorts. As observed in previous studies, avidity measures tend to be less affected by slight alterations in the assay or sample, given that avidity index is calculated relative to a control included within the same assay plate One of the most important considerations for a TRI is whether the recent/long-term classification is impacted by the use of DBS, thereby affecting incidence estimates. Since “true” recent/long-term status is not known for all of the samples included in this study, we compared the number of recent infections as determined by the assay values for plasma versus DBS. This evaluation was done strictly for demonstrative purposes, as finalized cutoffs have yet to be determined for the assay. Although demonstrated with a relatively small sample set, the number of specimens classified as recent were similar (differences ≤2) between plasma and DBS for all analytes except gp160-n. The number of specimens classified as recent differed between the analytes, given that each analyte has a distinct mean duration of recency (MDR) and, therefore, the determination of recent infection is analyte-specific Given that most TRI formats are laboratory-based, the use of DBS as an additional sample source is highly valuable since samples can be collected in resource-limited settings and shipped for later testing. We demonstrate stable reactivity of antibodies from DBS, even after specimens have been stored for several years. The modification to utilize DBS for the HIV-1 Bio-Plex assay described here expands its utility as a TRI for estimating HIV incidence.


However, the underpinning evidence has largely been obtained from studies of populations of white European descent. It is unclear whether these recommendations are appropriate for other ethnic groups, particularly South Asians, who have greater cardio-metabolic risk than white Europeans. The objective of our study was to determine the level of moderate-intensity physical activity required in South Asians adults to confer a similar cardio-metabolic risk profile to that observed in Europeans of similar age and body mass index (BMI) undertaking the currently recommended levels of 150 min.week−1. 148 South Asians and 163 white Europeans aged 18 to 70 years were recruited. Physical activity was measured objectively via vertical axis accelerations from hip-worn accelerometers. Factor analysis was used to summarize the measured risk biomarkers into a single underlying latent “factor” describing overall cardio-metabolic risk. Sex did not modify the association between physical activity and the cardio-metabolic risk factor, so data for both sexes were combined and models adjusted for age, sex, BMI and accelerometer wear time. We estimated that South Asian adults needed to undertake 232  min.week−1 in order to obtain the same cardio-metabolic risk factor score as a white European undertaking 150 minutes of moderate-equivalent physical activity per week. The present findings suggest that South Asian men and women need to undertake ~230 minutes of moderate intensity physical activity per week. This equates to South Asians undertaking an extra 10–15 minutes of moderate intensity physical activity per day on top of existing recommendations.International public health guidelines recommend that adults undertake at least 150 min.week Like obesity, a low level of physical activity is an important risk factor for T2D and CVD . Ho. Ho-2 isctivity) –25 and activity) ,27.The aim of this study was therefore to determine the level of physical activity required in South Asian women and men living in the UK to confer an equivalent cardio-metabolic disease risk profile to white Europeans undertaking the current guideline level of physical activity.Data on South Asian  and European women (both parents of white European descent) were collected from a cross-sectional study conducted in Scotland. Participants were recruited mainly through general advertising and word of mouth. Participants were not compensated for their time. Women aged between 18 to 70 years, lived in the UK and did not have a history of CVD, diabetes or polycystic ovary syndrome. Women on hormone replacement therapy or hormonal contraceptives were excluded. Only women with valid data for objectively monitored physical activity were included in this analysis. Data were combined with valid data from men that were recruited for the Carotid Ultrasound and Risk of Vascular disease in Europeans and South Asians (CURVES) study which has been described extensively elsewhere . ParticiBoth studies were approved by the West of Scotland Research Ethics Committee and conducted according to the Declaration of Helsinki. All participants gave written informed consent to participate.bouts). This was the second activity variable (exposure) for additional analysis.Participants wore accelerometers  at all times except when showering, swimming and sleeping for seven consecutive days. Vertical axis accelerometer counts were summarised into 60-second epochs and activity intensity domains were classified based on the Freedson cut-points . Valid dVenous blood samples were collected after an overnight fast. Glucose and glycated haemoglobin (HbA1c) were analysed as routine samples on the day of collection in one of the certified NHS Biochemistry laboratories within Greater Glasgow and Clyde by using standard automated enzymatic and HPLC techniques. Centrifuged serum and plasma were stored at -80°C for subsequent analysis. Lipids , triglycerides (TG) were measured in thawed sera using automated enzymatic technique at the end of the study. LDL levels were calculated with the use of the Friedewald equation . Insulin2  =  0.10)) in these analyses were clustered into summary factors. The single summary factor that accounted for the largest proportion of variation of the variables was selected as the dependent variable. This included the standardized levels of HbA1c, glucose, HDL-c, TG, insulin, total cholesterol, systolic and diastolic blood pressure. Regression models were fitted with the summary factor as dependent variable and either total moderate-equivalent physical activity or MVPAbouts as the exposure variables. In addition, all models contained the interaction of ethnicity with the independent variable or the interaction of sex with the independent variable. If the interaction terms did not improve the fitting of the model, then the interaction term was not included in the model and sex was included as a covariate. In addition, the models were adjusted for age, accelerometer wear time, BMI and smoking status. Statistical significance was adopted at p < 0.05.Statistical analysis was performed using STATA package . Using a similar approach described previously –20, fact311 out of 364 participants (85.4%) (n = 153 of 178 women (86.0%) and n = 158 of 186 men (85.0%)) had valid accelerometer data. There were no marked differences in baseline characteristics and key variables between the participants with valid and without valid accelerometer data (data not shown). bouts with the cardio-metabolic risk factor. Both associations were attenuated by adjustment for age, BMI, ethnicity, sex, smoking and total accelerometer wear time. However, while the association of total moderate-equivalent physical activity and the cardio-metabolic risk factor remained significant after adjusting for confounders, the association of MVPAbouts and the cardio-metabolic risk factor was substantially attenuated by adjustment and lost statistical significance. In the unadjusted models, total moderate-equivalent activity explained 18.4%, whereas MVPAbouts explained 12.6% of the variance in the summary cardio-metabolic risk factor. Therefore, total moderate-equivalent activity (rather than MVPAbouts) was used as the exposure in estimating the equivalent levels of physical activity to 150 min.week-1 for the South Asians. Overall, the fully adjusted model including total moderate-equivalent activity, age, BMI, ethnicity, sex, smoking and total accelerometer wear time variance in the cardio-metabolic risk factor explained 44% of the variance in the cardio-metabolic risk factor. BMI and smoking, the other modifiable risk factors included in the fully adjusted model, explained 8% and 2.4%, respectively, of the variance the cardio-metabolic risk factor in univariate models.Ethnicity and sex did not modify the associations between activity variables and the summary cardio-metabolic risk factor; hence data from all participants were included in the same model. -1 (95% CI: 200 to 268) of moderate-equivalent physical activity to achieve the same cardio-metabolic risk factor value as European adults undertaking 150 min.week-1 moderate-equivalent physical activity, in analyses adjusted for age, BMI and accelerometer wear time. There were no significant physical activity x sex, or physical activity x ethnicity x sex interactions with the cardio-metabolic risk factor outcome, indicating that the required level of physical activity in South Asians for equivalent risk to Europeans did not differ between men and women. The analyses suggest that South Asian adults need to undertake 232 min.week-1 versus 150 min.week-1 of moderate equivalent physical activity respectively). These data from a mixed-sex cohort extend our previous observations in men [This study demonstrates that South Asians of both sexes need to undertake higher levels of physical activity to exhibit an equivalent cardio-metabolic risk profile to that of adults of white European descent of similar age and BMI achieving current physical activity guidelines  are more likely to exhibit impaired glycaemia compared to women with self-reported weekly activity greater than this threshold (sensitivity 84% and specificity 85%) [-1 [-1 of physical activity performed in bouts of 10 min or 384 (231 to 536) min.week-1 of total moderate physical activity  in order to equate their overall cardio-metabolic risk with that of men of white European descent [-1. This level of activity is within the confidence intervals of the earlier analysis limited to men, and the larger sample size here enables the physical activity estimate to be made with greater precision than our previous analysis. However, direct comparison with the previous estimated recommended time for South Asian men cannot be made as we use a more adjusted model in our index analysis and our exposure is moderate-equivalent physical activity rather than moderate-to-vigorous physical activity or bouts of at least 10 minutes duration.The concept of ethnicity specific recommendation for physical activity has been discussed elsewhere. A recent study in Sri Lankan women demonstrated women who self-reported less than 2,640 MET-minutes.weekity 85%) . Althougity 85%)  and limi85%) [-1 , however descent . The preA key strength of this study is that physical activity was measured with the use of an objective measure which is essential in quantifying the real dose-response relationship between physical activity and cardio-metabolic risk. In contrast, self-report measures of physical activity have been shown to be imprecise and to substantially overestimate activity levels, which has the consequence of masking the true magnitude of the association between activity and risk . Althoug-1 was largely based on studies in which physical activity was self-reported, rather than objectively measured. Accelerometer-measured physical activity is typically lower than self-reported values—in one estimate by 2.5-fold [-1 of self-reported physical activity could conceivably be equivalent to ~60 min.week-1 of accelerometer-measured physical activity. Re-running our analysis using 60 min.week-1 as the guideline amount of physical activity, results in an equivalent estimate for south Asians of ~140 min.week-1. Thus, using either approach, south Asians would need to undertake ~80 min/week more (accelerometer measured) physical activity (232 mins vs 150 mins or 140 mins vs 60 mins) than white Europeans for a similar cardio-metabolic risk profile. This corresponds to ~15 minutes additional physical activity per day if individuals are active on 5 days of the week.It is important to recognize that the evidence base underpinning the current 150 min.week2.5-fold –thus 150-1 are warranted before definitive recommendations regarding ethnicity-tailored physical activity can be implemented.The present study has some limitations. It is cross-sectional with a relatively modest sample size assuming that increasing physical activity in South Asians will moderate their background cardio-metabolic risk. Hence, randomised controlled trials (RCTs) examining whether South Asians undertaking the proposed levels of physical activity would exhibit a similar cardio-metabolic profile with that of white Europeans of similar age and BMI exercising for 150 min.weekNevertheless, the present work makes an important contribution to the evidence-base for ethnicity-specific physical activity recommendations, by extending previous observations to women and by increasing the precision of the estimate for the level of physical activity required in South Asian adults for equivalent risk. We acknowledge that the South Asians in this study may not be representative of all South Asians living in the UK (i.e. we excluded people with diabetes and only included specific age groups). Whilst this would limit the ability to generalize prevalence rates (which is beyond of the scope of this study), estimates of the magnitude of associations between physical activity and CVD risk will not be affected by this and will therefore be generalizable. Thus, the recent recommendation from the Joint British Societies’ consensus recommendations for the prevention of cardiovascular disease that South Asian men may benefit from higher levels of physical activity than currently recommended , can reaThe present findings suggest that South Asian men and women need to undertake ~230 minutes of moderate intensity physical activity per week to confer a similar cardio-metabolic risk profile to adults of white European descent achieving current physical activity recommendations of 150 minutes per week. This equates to South Asians undertaking an extra 10–15 minutes of moderate intensity physical activity per day. While an RCT is needed to demonstrate whether these levels of physical activity would moderate the cardio-metabolic risk of South Asians, these data highlight the need of educating South Asians of the importance of physical activity and the development of culturally appropriate strategies to increase physical activity in this ethnic group.S1 Table(DOCX)Click here for additional data file.


Parvoviridae family, in particular the rodent protoparvoviruses H-1PV, the minute virus of mice and LuIII have natural anticancer activity while being nonpathogenic to humans. These studies have laid the foundations for the launch of a first phase I/IIa clinical trial, in which the rat H-1 parvovirus is presently undergoing evaluation for its safety and first signs of efficacy in patients with glioblastoma multiforme. After a brief overview of the biology of parvoviruses, this review focuses on the studies which unraveled the antineoplastic properties of these agents and supported their clinical use as anticancer therapeutics. Furthermore, the development of novel parvovirus-based anticancer strategies with enhanced specificity and efficacy is discussed, in particular the development of second and third generation vectors and the combination of parvoviruses with other anticancer agents. Lastly, we address the key challenges that remain towards a more rational and efficient use of oncolytic parvoviruses in clinical settings, and discuss how a better understanding of the virus life-cycle and of the cellular factors involved in virus infection, replication and cytotoxicity may promote the further development of parvovirus-based anticancer therapies, open new prospects for treatment and hopefully improve clinical outcome.Accumulated evidence gathered over recent decades demonstrated that some members of the  Parvoviridae family deserves special consideration for its promising anticancer properties. The RoPV viruses exert striking oncosuppressive effects in various preclinical tumor models, are able to kill tumor cells which resist conventional treatments, and have not been associated with disease in humans, laying the basis for the launch of the first phase I/IIa clinical trial using the rat oncolytic H-1 parvovirus (H-1PV).The idea to use viruses as tools for cancer therapy arose as early as at the turn of the 20th century, when it was reported that leukemia patients who contracted influenza went into clinical remission. Although anecdotal, these observations prompted intensive investigation of treatment strategies based on viruses with inherent anticancer activity, leading to the launch of the first clinical trials in the 1950s and 60s. Despite early promise, concerns regarding safety and a lack of efficacy caused a diminishment in oncolytic virotherapy research during the years that followed. However, with dramatic advances in our understanding of molecular biology and virology and the advent of genetic engineering, the last two decades have brought with them a resurgence of interest in the field , whose members are able to replicate autonomously in close dependence on cellular S-phase factors. RoPVs include the H-1 parvovirus (H-1PV), the major subject of this review, the Kilham rat virus (KRV), the LuIII virus, the Mouse parvoviruses (MPV) and the Minute viruses of mice (MVM). In unprotected fetuses and neonates of the natural or related hosts, RoPV infection can be pathogenic and even lethal, whilst in adults the infection is clinically inapparent though persistent. Interestingly, these viruses are able to replicate in cells of different origins, including transformed human cells, as it will be extensively discussed in the next paragraphs.The molecule . The famectively . Eight gThe RoPV capsid consists of 60 copies of two to three polypeptide sequences represented by the capsid proteins VP1, VP2 and VP3 . The capThe parvoviral genome is a linear, single-stranded DNA molecule of around 5 kb comprising two transcription units that respectively code for non-structural proteins (NS) involved in the replication process and in viral cytotoxicity, and structural proteins (VP) forming the capsid. The expression of viral genes is regulated by two promoters, P4 and P38 . P4 contThe parvoviral product NS1 is a multifunctional phosphoprotein of 672 amino acids with a theoretical molecular weight of 83 kDa. It is mainly located in the nucleus due to a nuclear localization signal (NLS) within its sequence , althougNS1 plays a major role in the RoPV life cycle, regulating both viral DNA replication and gene expression. It has been shown that NS1 is target for post-translational modifications that regulate protein functions. For instance, phosphorylation on different amino-acid residues catalysed by cellular protein kinases C (PKC), regulates both enzymatic (helicase and ATPase) and non-enzymatic  NS1 activities . FurtherBesides being involved in the regulation of the parvoviral life cycle, as will be discussed in the next paragraphs, NS1 is also the major effector of viral cytotoxicity and its sole expression is sufficient to induce cell cycle arrest and apoptosis -16.The RoPV protein NS2 has a molecular mass of about 25 kDa and a mainly cytoplasmic localization. In the case of MVM and related viruses, alternative splicing produces three different isoforms, NS2Y, NS2L and NS2P. The exact role of NS2 in the viral life cycle and cytotoxicity is still elusive. However, studies performed in mouse cells infected with MVM demonstrated a role of NS2 in the synthesis of DNA replicative forms and progeny production . FurtherProtoparvovirus genus [Recently, a novel non-structural protein has been identified, namely the small alternatively translated (SAT) protein, which is conserved among all members of the us genus . The opeus genus .2 (PLA2) activity, which is conserved among Parvovirinae members and thought to be required for the transfer of viral genomes from the endocytic compartment to the cytosol [The structural proteins VP1 and VP2 present the same nucleotide sequence except for the N-terminal domain: VP1 contains a unique sequence of 140 to 230 residues, besides the complete sequence of VP2. These additional residues have been shown to be important for viral infectivity but not for the packaging process ,20. The  cytosol -24. VP2  cytosol .Parvovirinae subfamily. For instance, the Feline and Canine parvoviruses, belonging to the genus Protoparvovirus, require the transferrin receptor for the entry process [Virus entry into target cells occurs through the interaction of the viral capsid with surface cellular receptors, which have been identified only for some members of the  process . Regardi process -29, thou2 activity of the VP1 protein triggers the degradation of the endosome by modification of its membrane composition, thereby favoring the release of viral particles into the cytoplasm [2 domain. In some experimental systems, Protoparvovirus members have proven able to induce disruption of the nuclear envelope through activation of key enzymes of mitosis [Protoparvovirus genome replication is strictly dependent on S-phase cellular factors. Therefore, incoming virions remain silent, without integrating their DNA into the cellular genome, until the host cell enters S-phase. As the cell expresses S-phase factors, viral DNA replication initiates. The replication process comprises the conversion of the single-stranded DNA molecule into a double-stranded replicative form. The newly synthesized double-stranded DNA molecules are used as templates for viral mRNA synthesis and for the formation of multimeric double-stranded intermediates and progeny DNA molecules [Viral capsids enter target cells through an endocytic mechanism, which is thought to involve clathrin proteins , and traytoplasm -24. The ytoplasm  and vira mitosis . Howeverolecules . Viral golecules ,36. Subsolecules ,37. Thesolecules ,38. It holecules ,39. NS1,olecules . This violecules .After the expression of the capsid proteins, new virions are produced and released, most likely through an active mechanism which brings the progeny virions from the nucleus to the cell periphery. Indeed, it has recently been reported that RoPV egress from the host cell occurs through the formation of virus-containing cellular vesicles in the endoplasmic reticulum, which are then transported to the membrane through the Golgi in a gelsolin-dependent manner . TranspoThe rodent protoparvoviruses MVM and H-1PV have the ability to induce cell cycle arrest in permissive cells - a process which is thought to favor viral replication ,16,38,42The components of the DNA damage machinery which relocate into the APAR bodies are present in their active phosphorylated form. The ataxia telangiectasia mutated protein (ATM), a key regulator of the DNA damage checkpoint, is the kinase involved in the phosphorylation of these factors, as treatment with specific ATM inhibitors reduces phosphorylation . ImportaH-1PV also has the ability to induce a DDR, which results in a G2/M cell cycle arrest. However the underlying mechanisms are less well characterized than for MVM. We have described that virus infection is associated with accumulation of reactive oxygen species (ROS). ROS, which are a known source of DNA damage, contribute, at least in part, to both virus-induced DDR and cell cycle arrest, as antioxidant treatment protected the cells from these events . InteresRodent protoparvoviruses can kill cancer cells through different mechanisms. Virus-induced apoptosis has been reported in several cell lines [Cell cycle arrest leads to cell death and, eventually, to cell lysis. ll lines ,16,48-51ll lines . In addill lines . Interesll lines ,54. As mll lines . The NS1ll lines .For efficient parvoviral multiplication to take place, it is important that the death of the target cancer cell does not precede virus genome replication and production of progeny virions. The NS1-mediated activation of the cellular PDK1/PKB survival pathway may be one of the mechanisms evolved by the virus in order to delay host cell death .Importantly, neither virus infection, nor the ectopic overexpression of the NS1 protein, is able to kill most normal cells ,58. Thisin vitro and in vivo studies have shown that these agents do not possess any tumorigenic activity but, on the contrary, exhibit the capacity to interfere with tumor growth [Although RoPV viruses were frequently isolated from tumor cells, r growth ,59. AmonRoPV oncoselectivity is not related to inefficient virus uptake by normal cells, but rather is due to the high dependence of the viral life cycle upon cellular factors that are dysregulated in tumor cells and involved in the control of cell proliferation and differentiation (see above).However, the presence of cellular helper functions is not the only pre requisite that determines the permissiveness of target cancer cells for RoPV infection. Indeed, the inactivation of antiviral immune mechanisms may also contribute to the efficacy of virus infection. In mouse cells, MVM proved to be sensitive to type I interferon-mediated defence mechanisms . Cancer As for other oncolytic viruses, the antineoplastic effects of RoPVs are due to direct oncolysis and/or to indirect immune reactions activated upon virus infection. These two events are not mutually exclusive, but most likely cooperate to achieve virus-mediated oncosuppression.in vivo studies, performed in immunocompetent rats bearing two pancreatic tumors implanted at distant sites, showed that, when H-1PV was injected at only one tumor site, regression of the tumor located at the distal site could be observed. This bystander effect is believed to be mediated by the immune system as a result of its virus-dependent activation at the injected site, in keeping with the accumulation of granzymes and perforin at the distal tumor site [in vitro [H-1PV infection of tumor cells stimulates the expression of danger- and pathogen-associated molecular patterns (DAMPs and PAMPs) and the release of tumor-associated antigens, thereby promoting the cross-presentation of tumor antigens by immature dendritic cells and facilitating the recognition of the tumor by the immune system . The immmor site . The immmor site . The indin vitro . AltogetOn the other hand, an antiviral humoral immune response, and more specifically a production of virus-neutralizing antibodies, was detected in animal models within one week after infection with H-1PV, MVM or the Mouse parvovirus MPV-1 ,71,72. Nin vitro and in vivo tumor models, as summarized in Table (i)Gliomain vivo. Upon intratumoral or intravenous delivery into rat orthotopic models of rat or human gliomas, H-1PV was able to induce complete tumor regression and to increase animal survival without causing inflammation or alteration of surrounding normal tissues [in vitro evidence that H-1PV induces a lysosomal type of glioma cell death [Glioblastoma multiforme (GBM) is the most common and aggressive type of primary brain tumor in humans, for which the median survival is 15 months after initial diagnosis . New the tissues . Virus ill death . Intranall death .Protoparvovirus members, including H-1PV, LuIII and MVM, were compared for their efficacy in killing a panel of glioma-derived cell lines. LuIII and H-1PV showed the greatest oncolytic activity when cells were infected with high multiplicities of infection (MOIs), while LuIII displayed superior anticancer activity when viruses were used at low concentrations, with an ability to propagate efficiently in all glioma cell lines tested. Interestingly, LuIII reduced tumor growth in mouse xenograft models when administrated both intratumorally and systemically, with no apparent adverse side effects for the animals [Interestingly, a combination approach using ionizing radiation and H-1PV showed increased cytotoxicity in a set of glioma-derived cell lines, including radio-resistant cells, compared to treatment with single agents. This effect could be observed more particularly when infection was performed after irradiation, and was associated with an increase in both NS1 levels and S-phase cell fraction. Importantly, this study demonstrated that previous exposure of glioma cells to radiation does not affect their susceptibility to H-1PV infection, supporting the potential clinical use of H-1PV in pre-irradiated glioma patients . Recentl animals .(ii) Neuroectodermal tumorsH-1PV oncolysis has also been reported in paediatric neuroectodermal tumors, such as neuroblastoma and medulloblastoma. Neuroblastoma is a highly malignant solid tumor originating from the central nervous system and is characterized by N-myc gene amplification, which is a biomarker of poor prognosis. Screening 11 neuroblastoma-derived cell lines for H-1PV sensitivity showed oncolytic effects in all the lines tested, irrespective of their p53 mutation or N-myc amplification status. Cytotoxic effects were associated with efficient viral replication, gene expression, production of progeny virions and induction of cell cycle arrest and apoptosis . InteresMedulloblastoma is the most common malignant brain tumor in children, while in adults its occurrence is rare. The potential of oncolytic virotherapy for the treatment of medulloblastoma is supported by the ability of several oncolytic viruses  to efficiently target this tumor type ,83. In a(iii) Lymphomain vitro and in vivo studies have demonstrated the ability of the virus to efficiently kill Burkitt’s lymphoma-derived cell lines (most likely through a necrotic mechanism and irrespective of EBV positivity) and to induce complete tumor destruction in severe combined immunodeficient (SCID) mice, accompanied by viral replication and spreading [in vitro; interestingly, oncolytic virus infection of these cells was productive in most cases .Burkitt’s lymphoma is a sporadic form of lymphoma, which is often associated with Epstein-Barr virus (EBV) infection. Although current treatment options led to improved survival in adults, an efficient cure is not yet available and, as a result, innovative treatments are needed. In this regard, H-1PV may be a potential candidate since preading . Further(iv) MelanomaKilling effects have been observed in human melanoma-derived cell lines infected with H-1PV, indicating that the virus may also be able to target this tumor type. H-1PV-induced melanoma cell death was found to be immunogenic, leading to the activation of antigen-presenting cells and tumor-specific cytotoxic T lymphocytes .(v)Mammary carcinomain vitro and in a xenograft mouse model [Breast cancer is another potential tumor targetable by H-1PV, as shown both se model ,87. Furtse model -89.(vi) Pancreatic carcinomaA number of studies have investigated the therapeutic potential of H-1PV against pancreatic ductal adenocarcinoma, a cancer type classified among the most frequent cause of cancer-related deaths, and for which effective treatment options are still lacking. Gemcitabine is a chemotherapeutic drug currently in use for the treatment of advanced local and metastatic pancreatic cancer. Unfortunately, resistance of cancer cells to gemcitabine dramatically reduces the efficacy of the therapy and causes tumor recurrence . H-1PV c(vii) Cervical carcinomain vitro and in vivo, with increased induction of oxidative stress, DNA damage and apoptosis, as well as with enhanced viral DNA replication, gene expression and production. The efficacy of the H-1PV/VPA combination was also demonstrated in a range of pancreatic cancer-derived cell lines and rodent xenograft models of human pancreatic carcinomas, indicating that this combination may represent a promising approach also for the treatment of this type of cancer.Cervical cancer is the second most common cancer in women. Although prevention screenings have significantly reduced mortality, and the vaccination program against human papillomaviruses, the major cause of cervical cancer, is expected to reduce tumor incidence among young women, the prognosis for the advanced disease remains poor, with a 1-year survival of only 15-20% . H-1PV i(viii)Gastric carcinomaH-1PV-induced killing of human gastric cancer cells was found to follow the up- or downregulation of the expression of a number of cellular genes . Recombi(ix) HepatomaEx vivo or in vivo infection with H-1PV also inhibited tumor development from human hepatoma xenografts in nude mice [H-1PV oncolytic effects have been reported in hepatoma-derived cancer cell lines, in which virus infection was associated with viral DNA replication, gene expression and virus production. Importantly, no cytotoxicity was observed in H-1PV-infected normal primary hepatocytes . Upon inude mice .(x)Colon carcinomaIn comparison with the tumor types described above, human colon cancer appears to exhibit lower susceptibility to the oncolytic activity of H-1PV, as suggested by the fact that infected colon carcinoma-derived cell lines show little cytopathic effects and fail to sustain efficient viral replication . HoweverIn this paragraph, special emphasis will be given to H-1PV that has been evaluated extensively at the preclinical level as an anticancer agent using a number of different The preclinical evidence obtained in glioma cultures and animal models provided the basis for the launching of a first phase I/IIa clinical trial, named ParvOryx01, involving patients with GBM.9 infectious units per patient. Biological monitoring is conducted by analysing the patients’ bodily fluids and resected tumor. The results of this accompanying research are intriguing, showing that H-1PV achieves widespread infection of gliomas, also after intravenous administration, and undergoes at least limited replication and expression in neoplastic tissues. Interestingly, this virus replication in some tumors is associated with necrosis and massive infiltration with T cells, including CTLs, some of which appear to be specific for viral and glioma epitopes (unpublished results). We trust that the further analysis of these surrogate markers of efficiency, as well as clinical data, will warrant the future extension of parvovirus clinical studies to subsequent phases, including also second-generation RoPV-based (combination) treatments.Besides the above-mentioned preclinical evidence of H-1PV anti-glioma effects, the ability of H-1PV to cross the blood-brain barrier and infect intracerebral tumors after systemic administration to rats gives hope for the development of non-invasive, H-1PV-based treatments of GBM . The trivβ3 and αvβ5 integrins, which are typically overexpressed on cancer cells and angiogenic blood vessels, and was used as a model ligand for virus retargeting to these cells. Indeed, the RGD-4C-decorated virus showed increased specificity for cancer cells, while retaining the oncolytic potential of the parental virus [in vitro and in vivo screenings.In recent years, H-1PV has been the subject of genetic manipulations that aimed at increasing virus oncospecificity and anticancer efficacy in order to optimize the therapeutic potential of RoPV-based treatments. Genetic engineering of the H-1PV capsid proved to be a suitable approach to increase virus specificity for cancer cells at the level of cell recognition and entry. The above-mentioned determinants of RoPV oncotropism indeed operate at the intracellular level. In consequence, the wild-type virus is able to enter most normal cells, which undergo a cryptic or abortive infection that is clinically unapparent but results in the sequestration and loss of a major fraction of inoculated virions away from the tumor target. In a first step, sites were identified on the capsid, which were involved in binding to normal cellular receptors, and whose modifications led to virus detargeting, i.e. lack of uptake by normal cells. In a second step, a site was identified within the viral capsid tolerating the insertion of the RGD-4C cyclic peptide. This peptide recognizes αal virus . This stal virus ) or by iin vivo [In another approach, the H-1PV genome was inserted into a replication-defective adenovirus vector genome, with the aim of generating an adenovirus-parvovirus chimera. The chimeric vector multiplied to high-titers, thus allowing the large-scale production of parvoviral DNA-containing particles, which is generally difficult to achieve with current parvovirus production procedures, rendering the chimera virus more suitable for clinical applications. The parvoviral genome was efficiently released from the adenoviral DNA backbone in target cancer cells, leading to autonomous H-1PV replication, cytotoxic effects and production of fully infectious parvoviral particles. The adenoviral carrier allowed H-1PV to be delivered into tumor cells originally resistant to direct parvovirus entry, and to kill these cells under conditions in which neither parental virus was efficient. While showing this broader oncolytic activity, the adeno/parvoviral chimera kept the innocuousness of its infectious parvovirus component for normal cells, thereby warranting further (pre)clinical assessment in vivo . Future Interestingly, the immunomodulatory effect of H-1PV could be enhanced by arming the virus with specific PAMPs, namely CpG motifs . It remaIt has been observed that cancer cell lines differ in their sensitivity to RoPV infection, with some of them being poorly permissive to virus replication and cytotoxicity. RoPV replication in human cancer cells can be enhanced through virus adaptation. Isolation of virus variants adapted for efficient production and spreading in human glioma cell cultures was recently achieved by serial passaging H-1PV in semi-permissive glioma cells .As mentioned above, combinations of H-1PV with physical (ionizing radiation) and chemical  cytotoxic agents, or with epigenetic modulators (HDAC inhibitors) resulted in striking synergistic oncolytic effects both in cell culture and animal models, providing the preclinical proofs of concept needed to move these protocols into the clinic. We anticipate that future studies will be directed toward the identification of other anticancer agents or therapeutic modalities (e.g. immunotherapy) that may act additively or synergistically with the virus in eliminating cancer cells, reinforcing the antineoplastic activity of the virus whilst keeping its excellent safety profile . This advance may also allow a reduction of the therapeutic dose to be used in cancer patients while maintaining or even enhancing efficacy.in vitro indicates that further optimisation of RoPV-based therapy is likely to be critical for the treatment of advanced human cancers characterised by high intratumoral heterogeneity and genomic instability.Oncolytic viruses are gaining momentum as a novel form of anti-cancer therapy. As demonstrated in the numerous pre-clinical models described above, RoPVs display striking oncosuppressive effects whilst being non-pathogenic to humans. For the full potential of these anti-cancer agents to be exploited, we believe that the development of more complex treatment strategies, aimed at enhancing virus replication, direct oncolytic activity and/or immunological adjuvant effects, will be necessary. Indeed, the varying susceptibility of tumor cell lines to RoPV replication and cell killing observed To this end, we anticipate that combination therapy using agents that synergize with RoPVs, may hold the key for maximizing anti-cancer effects. The permissiveness of a given cancer cell for RoPV infection and replication is governed by an intricate set of virus/host interactions which, as is the case with many OVs, remain largely unknown. A more thorough understanding of the parvoviral life-cycle and host factors implicated in it may reveal novel targets for therapeutic intervention, thus guiding the development of RoPV-based combination therapies with augmented anti-neoplastic activity. The power of such rationally designed combinations to reinforce RoPV therapy is exemplified by the synergism observed between H-1PV and HDAC inhibitors, following the discovery that acetylation of NS1 enhances its cellular functions . To effiThe above-mentioned rational design of combination therapy requires prior knowledge of particular virus host interactions. As an alternative approach, high-throughput screening of drug and chemical libraries may be performed in an unbiased fashion to identify compounds that potentiate OV therapy. This approach has already shown promise as an efficient way to discover novel enhancers of OV activity. For example, equilibrative nucleoside transporter-1 (ENT1) antagonists were found to be potent amplifiers of oHSV replication following a drug screening to identify molecules that increased viral spread , whilst in vitro, their safety and efficacy must be corroborated in vivo. Pre clinical validation of RoPV-based therapies is traditionally performed using two classes of in vivo systems: (i) xenograft models, generated from the implantation of cultured human tumor cells into immunocompromised rodents, and (ii) immunocompetent syngeneic models, bearing tumors originating from the same species. Despite providing valuable indications of efficacy, both approaches carry significant drawbacks that limit their utility as predictive tools of clinical outcome: xenograft models will not reveal the positive or negative consequences of the host immune response to treatment, whilst results from syngeneic animal models can be misleading since the behavior of the virus within cells, as well as immune response towards it, may vary considerably between species. Furthermore, tumors established from cultured cell lines seldom recapitulate the complexity or heterogeneity of those observed in patients. For predicting the clinical outcome of novel RoPV-based treatments and selecting the best combination therapies to move towards the clinic, patient-derived tumor xenograft models are likely to be more informative as they retain greater similarity to parental tumors [Following the identification of promising PV-drug combinations l tumors . Considel tumors . Reconstl tumors , making l tumors .Another significant challenge facing the field, which may hamper the successful translation of these therapeutics to the clinic, is the large-scale production of RoPV-based vectors in compliance with Good Manufacturing Practice (GMP). To produce RoPV vectors at the quality and quantity required to meet clinical demand, major efforts must be directed towards the search for more productive packaging cell lines and optimisation of culture media and growth conditions, as well as the improvement of purification procedures. The development of high-yielding, scalable bioreactor platforms using cell lines growing in suspension is also an important goal for reducing the considerable costs of virus production.Looking forward, we believe that the identification of biomarkers predictive of sensitivity/resistance to RoPV therapy will also be a critical step towards its effective use in clinical settings. With this knowledge, patients may be screened for the presence of specific markers and selected for “smart” clinical trials based upon their likelihood of responding to treatment. Moreover, in those patient subsets exhibiting certain features that could conceivably compromise efficacy, outcome may be improved through the rational application of combination therapy and/or the use of engineered RoPVs. Figure The successful translation of laboratory results into effective cancer treatments requires more than just talented researchers and inspired clinicians. Early-phase trials of oncolytic viruses are typically conducted on patients with advanced or recurrent cancers, for whom all other therapeutic options have failed. However, conventional first-line treatments of some forms of cancer, including glioblastoma multiforme, provide only modest benefit to the patients and are often associated with severe adverse side-effects. Furthermore, the ability of patients to mount a robust immune response is often compromised at such late stages of disease, which may prevent them from benefiting from the immunostimulating action of RoPVs. Long-term therapeutic improvements may therefore be gained primarily by patients treated at early stages of their disease. This raises a delicate regulatory issue: should patients with aggressive cancers, for which no curative therapies are currently available, have access to novel treatments at an earlier point after diagnosis? While lying beyond the scope of this review, this regulatory issue deserves more discussion.Exciting results at the pre clinical level offer a tantalizing glimpse of the potential of oncolytic parvoviruses as novel anti-cancer agents. However, the successful translation of these promising laboratory results into clinically effective treatments is still in its infancy. We anticipate that the ongoing clinical trial will provide valuable information enabling the further exploitation of RoPV therapeutic properties, as well as revealing hurdles that may limit efficacy. Overcoming these limitations represents a challenge to be taken up first at the bench level. Guided by better understanding of the cellular factors controlling RoPV-host interactions and malignant progression, we endeavor to develop next-generation RoPV-based treatments with improved anticancer efficacy. This effort will hopefully contribute to reinforce the impact of RoPVs on the fight against this devastating disease.Oncolytic virotherapy of cancer has emerged as a promising alternative to toxic chemotherapy regimens. Among a dozen oncolytic viruses presently tested at the clinical level, rodent parvoviruses (and the H-1 parvovirus in particular) attract considerable attention, due to their remarkable natural oncoselectivity and lack of pathogenicity for humans. In the last two decades, an impressive amount of preclinical data has been accumulated, showing that H-1PV possesses both oncolytic and immunostimulating properties. These give the virus a significant cancer therapeutic potential, either alone or in combination with other agents (e.g. histone deacetylase inhibitors), as revealed in several tumor models. Cancers targeted by RoPVs include glioblastoma multiforme, the most common and aggressive primary brain tumor in humans. The first phase I/IIa clinical trial using an oncolytic parvovirus (H-1PV) was launched in 2011 for patients with recurrent GBM. We trust that this ongoing clinical study will pave the way for subsequent efficacy trials, and prompt the development of next-generation parvovirus-based anticancer therapeutics.


Accumulating evidence suggests that Japanese herbal medicines, called Kampo, have beneficial effects on cancer chemotherapy-induced side effects. Rikkunshito ameliorates cisplatin-induced anorexia through an antagonistic effect on the 5-HT receptors and by increasing the serum ghrelin levels. Hangeshashinto improves irinotecan-induced diarrhea and chemotherapy-induced mucositis by inhibiting the activity of β-glucuronidase as well as the synthesis of prostaglandin E2. Goshajinkigan prevents oxaliplatin-induced neurotoxicity, possibly through suppressing functional alterations of the transient receptor potential channels. In this review, we will summarize the currently available literature regarding the clinical efficacy and potential mechanisms of Kampo medicines in the treatment of cancer chemotherapy-induced side effects. Cancer is a major public health problem in most developed countries; however, there have been notable improvements in the survival rate of patients over the past three decades owing to early detection and progress in medical treatment . A substJapanese traditional herbal medicines, called Kampo, were imported from China 1,500 years ago and developed independently . Kampo mIt is important to prevent and treat chemotherapy-induced side-effects such as anorexia, nausea, and vomiting to maintain patients’ QOL and to continue chemotherapy safely . Nausea International guidelines for antiemetic use have recommended antagonists for 5-HT3 and NK-1 receptors as well as corticosteroids for patients undergoing treatment with anticancer drugs including cisplatin . HoweverGhrelin is a GH-releasing peptide isolated from the stomach as an endogenous ligand for GH secretagogue receptor 1a GHS-R1a; , a G-proRikkunshito is composed of 8 herbal medicines and is widely used in Japan to treat various gastrointestinal disorders . ThroughSeveral clinical trials have been conducted to investigate the effect of rikkunshito on chemotherapy-induced anorexia. CPT-11 exerts an inhibitory effect on nucleic acid synthesis by inhibition of topoisomerase I and is widely used for the treatment of a variety of cancers . One of 2 of CPT-11 suffered from grade 3 diarrhea.Hangeshashinto is composed of seven herbs and is often used in Japan to treat diarrhea and acute gastroenteritis . HangeshOral mucositis is a common complication of chemotherapy, affecting 18–40% of patients after the first chemotherapy cycle . The morN = 14), topical application of hangeshashinto improved oral mucositis (Hangeshashinto contains several prostaglandin E2-regulating ingredients . In a reucositis . Aoyama Oxaliplatin, a platinum compound, is widely used for the treatment of various cancers, mainly colorectal cancer. However, oxaliplatin-induced neurotoxicity is the most common dose-limiting side effect . AlthougGoshajinkigan is composed of 10 herbal medicines and is widely used in Japan for the treatment of rhigosis, numbness or pain in the extremities, low back pain, and diabetic neuropathy . Animal Growing evidence suggests that Kampo medicines appear to have beneficial effects for the treatment or prevention of several chemotherapy-induced side effects. Because each Kampo medicine is not composed of a single substance, further basic studies should clarify which of the substances are responsible for the beneficial effects as well as the underlying mechanism of action. Although it is sometimes difficult to set appropriate and subjective endpoints for Kampo medicines, placebo-controlled, double-blind, randomized clinical trials similar to those for Western medicine will provide decisive evidence.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Hiroshi Takeda received grant support from Tsumura & Co. Shunsuke Ohnishi declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


These glycans can be carried on many mucin-type glycoproteins including MUC1. We show that the majority of breast cancers carry Tn within the same cell and in close proximity to extended glycan T  the addition of Gal to the GalNAc being catalysed by the T synthase. The presence of active T synthase suggests that loss of the private chaperone for T synthase, COSMC, does not explain the expression of Tn and STn in breast cancer cells. We show that MUC1 carrying both Tn or STn can bind to the C-type lectin MGL and using atomic force microscopy show that they bind to MGL with a similar deadadhesion force. Tumour associated STn is associated with poor prognosis and resistance to chemotherapy in breast carcinomas, inhibition of DC maturation, DC apoptosis and inhibition of NK activity. As engagement of MGL in the absence of TLR triggering may lead to anergy, the binding of MUC1-STn to MGL may be in part responsible for some of the characteristics of STn expressing tumours.Aberrant glycosylation occurs in the majority of human cancers and changes in mucin-type O-glycosylation are key events that play a role in the induction of invasion and metastases. These changes generate novel cancer-specific glyco-antigens that can interact with cells of the immune system through carbohydrate binding lectins. Two glyco-epitopes that are found expressed by many carcinomas are Tn  and STn (NeuAc Glycosylation is one of the most widely found and complex post-translational modifications, and the glycome encompasses a vast and extensive repertoire of sugars covalently linked to proteins, glycolipids or proteoglycans. The mammalian glycome is estimated to contain thousands of different glycan structures, vastly expanding the diversity of the proteome, and is involved in key biological processes. Nearly all proteins that are expressed on the cell membrane, or are secreted, carry glycans and these are involved in cell adhesion, recognition, molecular trafficking, clearance and signalling . Indeed,β3galactosyltransferase, also known as T synthase. The activity of T synthase is totally dependent upon a private molecular chaperone known as Cosmc ..24].in vivo situation [int), to 85 and 81 respectively  upon increasing the density of MUC1-ST on the AFM tip  was used see . The fulituation . The intvely see . The cor tip see .The distribution of the de-adhesion work for the MUC1-Tn and MUC1-STn-MGL interactions is displaced towards larger de-adhesion work compared to the MUC1-ST-MGL interaction see . This reThe weak correlation between the amount of interaction and the density of the MUC1-ST indicates that the adhesion events observed for this sample is not due to a specific MUC1-ST–MGL molecular bond but to unspecific adhesion between AFM tips and mica surface. Such unspecific adhesive interactions were previously quantified in our laboratory . The posTo confirm the specificity of the interactions, the effect of adding of adding Tn-Ser to MUC1-Tn and STn-Ser to MUC1-STn interactions with MGL was studied. Upon addition of Tn-Ser or STn-Ser to the buffer, a shift towards reduced de-adhesion work was observed both for the MUC1-Tn and the MUC1-STn systems. in vivo around 25% of breast carcinomas express STn, few cell lines express this glycan. We therefore made use of T47D cells transfected with ST6GalNAc-I, which results in the expression of STn on the cell surface [COSMC knocked-out via gene editing [Having demonstrated that MUC1-Tn and MUC1-STn show a similar capacity to interact with MGL, it was important to show an interaction between cell surface MUC1-STn and MGL. Although  surface , see Fig editing  to invesChanges in glycosylation are common events in cancer and can lead to aberrant interactions of glycoproteins expressed by the tumour cell with glycan binding proteins, including C-type lectins expressed by normal cells. In the case of binding to lectins on immune cells this may induce the secretion of inflammatory mediators that are believed to drive tumour growth and progression or may induce the uptake of soluble glycoproteins leading to antigen presentation ,1.COSMC is still active in these breast cancer cells. Moreover in the large METABRIC study, expression of COSMC was in fact increased in breast cancer compared to normal  [COSMC expression by DNA methylation may not play a role in dictating the expression of Tn and STn in breast cancer, although this can only be confirmed by determining the T synthase activity.Breast cancer can carry the truncated glycans Tn and sialyl-Tn (STn). One mechanism that may result in O-linked glycoproteins carrying these truncated glycans is the inactivation or lack of expression of COSMC. As mucin O-glycans are synthesised by the sequential addition of individual sugars, inactivation of COSMC, resulting in a non-functional T synthase, blocks further chain elongation so that O-glycosylation is truncated to Tn or STn if ST6GalNAc-I is active, and T glycoforms should not be detected . In the 5.15E-6)  suggesti5.15E-6) , silenciWhen glycans are carried on MUC1 they can be repeated hundreds of times. Depending on the allele, the extracellular domain of MUC1 consists mainly of 25–150 tandem repeats of 20 amino acids. Each repeat carries five O-linked glycosylation sites, thus glycans can potentially be repeated 125–625 times on each molecule allowing engagement and crosslinking of relevant lectins. To begin to understand the implications of these truncated glycans being carried on MUC1 we focused our studies on MGL binding. Human MGL was thought to exclusively recognise GalNAc ,36, howeLectins expressed by immune cells have been thought to be involved in the recognition of PAMPs (pathogen associated patterns) although it is now clear that they can also bind host glycans. Indeed lectins are involved in the recognition of DAMPs (danger associated molecular patterns) released during tissue damage  and it hα secretion [MGL has been reported to be a marker of alternative activation in murine models of allergy and parasitic infection, with IL-4 and IL-13 being shown to be crucial for its induction and maintenance . Indeed,ecretion ,46.Whilst STn is a terminal glycan, the expression of surface terminal GalNAc structures is unusual in breast cancer as it must be a result of incomplete O-linked glycosylation as our data rules out the cause being lack of COSMC function. Indeed the Tn glycan is mostly intracellular and not frequently found on the surface of carcinomas . Thus thS1 Fig1μg of recombinant MUC1-Tn or MUC1-STn was coated onto each well of a 9well dish. After washing 5E5 was added and the binding visualized with rabbit anti-mouse IgG peroxidase conjugated.(PPTX)Click here for additional data file.S1 TableResults of the IHC of the individual tumours.(DOCX)Click here for additional data file.


Staphylococcus aureus (MRSA). The QAC being tested is an antibacterial film that is sprayed onto a surface and can remain active for up to 8 h. A field experimental study was designed with the QAC plus daily hypochlorite cleaning as the experimental group and hypochlorite cleaning alone as the control group. The method of swabbing on moistened surfaces was used for sampling. It was found that 83% and 77% of the bedside surfaces of MRSA-positive and MRSA-negative patients respectively were contaminated with staphylococci at 08:00 hours, and that the staphylococcal concentrations increased by 80% at 1200 h over a 4-hour period with routine ward and clinical activities. Irrespective of the MRSA status of the patients, high-touch surfaces around the bed-units within the studied medical ward were heavily contaminated (ranged 1 to 276 cfu/cm2 amongst the sites with positive culture) with staphylococcal bacteria including MRSA, despite the implementation of daily hypochlorite wiping. However, the contamination rate dropped significantly from 78% to 11% after the application of the QAC polymer. In the experimental group, the mean staphylococcal concentration of bedside surfaces was significantly (p < 0.0001) reduced from 4.4 ± 8.7 cfu/cm2 at 08:00 hours to 0.07 ± 0.26 cfu/cm2 at 12:00 hours by the QAC polymer. The results of this study support the view that, in addition to hypochlorite wiping, the tested QAC surfactant is a potential environmental decontamination strategy for preventing the transmission of clinically important pathogens in medical wards.The aim in this study was to assess the effectiveness of a quaternary ammonium chloride (QAC) surfactant in reducing surface staphylococcal contamination in a routinely operating medical ward occupied by patients who had tested positive for methicillin-resistant  Microorganisms on hospital surfaces can be transmitted to the hands of healthcare workers, patients, and visitors, resulting in cross-infections and epidemics. Despite the implementation of routine cleaning and precautionary measures in most hospitals, effective environmental decontamination methods are still in demand. In recent decades, numerous polymeric surfactant products have been shown to have excellent antimicrobial properties against surface contamination, but none have been tested on hospital surfaces ,2. JUC sIn hospitals, surfaces with which patients have close contact or that are highly accessible to patients are more likely to become contaminated. Environmental MRSA contamination has been extensively reported in different areas of a hospital, including in intensive care units ,7,8,9, bS. aureus from Hong Kong hospitals were shown to be methicillin resistant [The prevalence of hospital-associated MRSA (HA-MRSA) infection varies geographically. Hong Kong is one of the high-prevalence areas in Asia . Accordiesistant . In Hongesistant . ProtectA field experimental study was designed to assess the degree of staphylococcal contamination on the bedside surfaces of a medical ward, and the effects of JUC polymer on reducing such contamination. Routine operations continued to be carried out in the ward to reflect the most natural environment for assessment. The study hospital was a large teaching hospital in Hong Kong with a capacity over 1500 beds. The study ward was a male medical ward, consisting of a total of three cubicles each containing six beds, which were fully occupied on all of the days during the study period. The ward and cubicle setting is illustrated in 2 per sampling site) at all of the selected sites. Using flocked or macrofoam swabs were found to be a reliable and efficient strategy for recovering bacteria from environmental surfaces [® sterile macrofoam tip environmental swab applicators for swab sampling. Swabbing was performed at three random spots of each sampling surface standardized to 10 square centimeters. Hence, a total of 12 swab specimens (three samples per site × four spots) were collected from each bed-unit. The swabbed specimens were inoculated into vials containing 1 mL of sterile water and transferred to PetrifilmTM Staph Express Count Plates .Four high-touch bedside surface sites  were selected for the swab sampling. All of the selected sites had been identified as high-touch surfaces using a quantitative approach . The samsurfaces . In thisThe degree of contamination in the bed-units occupied by MRSA-positive and MRSA-negative patients was compared. Swab specimens were collected from all bed-units within the cubicle at two time points at 08:00 hours and 12:00 hours, and this was repeated on six separate days over a 6-week period. The time of eight o’clock in the morning was chosen because that is not long after hypochlorite wiping has been done, at 07:00 hours, and most patients are awake following the minimal activities during the night shift within the ward. The hypochlorite solution was freshly prepared according to hospital guidelines by a duty healthcare worker, who also performed the wiping of the whole cubicle under the supervision of a duty nurse to ensure that all surfaces were cleaned. Hence, the 08:00 hours specimens were used as the baseline for comparisons between the MRSA-positive and MRSA-negative surfaces. Furthermore, the 12:00 hours specimens were used to assess natural changes in contamination over the four-hour interval during which routine clinical activities were carried out without disturbance. During the four-hour period before the lunch hours, visitors are generally not allowed to enter the ward, which minimizes disturbances other than those from patients and healthcare workers. However, healthcare professionals outside the ward cannot be prevented from entering the ward. Over the course of the 6-week study, a total of 864 swab specimens (three samples per site × four spots × six beds × two time points × six days) were collected from the designated site surfaces.i.e., sanitization with bleaching water at 07:00 hours), while the control arm tested the routine daily cleaning alone. Regardless of the study arm, swab specimens were collected at 08:00 hours as the baseline and at 12:00 hours for assessing the change in the staphylococcal burden. The JUC dressing spray was gifted by the NMS Technologies Company . On the experimental days, the JUC spray was applied to the site surfaces immediately after the baseline specimens had been collected . To avoid a carry-over effect from the JUC spray, each experimental day was arranged to be at least 7 days apart from the next experimental day. The control arm was also conducted on three separate days. On each experimental and control day, one MRSA-positive patient occupied the bed-unit at the far end, i.e., Bed-unit A1 in In this experiment, the decontamination property of the JUC polymer was evaluated on the same designated sites of the bed-units used for the staphylococcal assessment in the same ward cubicle. The experimental arm tested the JUC spray plus the routine daily cleaning (cted see b. The fo2). Since each swab sample was collected from a surface area of 10 cm2, the CFU/Petrifilm actually represented the growth of staphylococcal colonies from each swab specimen covering 10 cm2 surface area. The minimum detection (countable) limit of the sampling technique was defined as 1 cfu/cm2. Positive growth on the plates was followed up using the coagulase test for differentiating coagulase-positive and coagulase-negative staphylococci (CNS). Subculture of the coagulase-positive staphylococci (red-violet colonies were randomly picked from Staph Express Count Plates) was performed in Mueller-Hinton agar supplemented with 4% w/v sodium chloride, and their sensitivity to oxacillin was assessed by using the disc diffusion test as recommended by the U.S. Centers for Disease Control and Prevention (http://www.cdc.gov/HAI/settings/lab/lab_mrsa.html). MRSA strains are resistant to all β-lactam antibiotics, but typically oxacillin resistant. Therefore, in this study, methicillin-resistant Staphylococcus aureus (MRSA) strains were identified and differentiated from the methicillin-sensitive Staphylococcus aureus (MSSA) strains. According to the technical bulletin and user instruction of the JUC spray dressing, the QAC solution will be solidified immediately after contacting the air after spraying onto any surface, and its bactericidial property is exerted by the physical electrostatic force generated between the positively-charged coating surface and the negatively-charged cell wall or membrane of the organisms. The coating adhered on the surface will then be water-proof and stay for up to 8 h. Such coating should not be removed from the sampling surface when performing the swab. Furthermore, physical bactericidal activity will be lost even if the coating is being released in solution, and therefore, neutralizer is not required for the specimen collection procedure from the bedside surface with the JUC coating and residual QAC is not a concern that could affect the microbiological results.Within one hour following the collecting of samples, all inoculated Staph Express Count Plates were sent to the microbiology laboratory of the study hospital for incubation and further analysis. All of the plates were incubated at 37 degrees Celsius for 24–48 h. Red-violet colonies were counted as positive for staphylococcal growth and expressed in Colony-forming Units per Centimeter Square  heavier than the amount recovered from the MRSA-positive bed-units. Overall, the staphylococcal distribution among CNS, MSSA, and MRSA was in the ratio of 2:6:8. Reference to the layout of the testing ward cubicle . Bed 4 was also observed to have relatively heavy growth. However, none of the beds had growth that was statistically different from that of other beds (data is not shown). Among all control bed-units received only hypochlorite cleaning, the mean staphylococcal contamination increased significantly (p < 0.01) by 80% from 08:00 to 12:00 hours  and 77% (23 out of 30) of the bedside surfaces occupied by MRSA-positive and MRSA-negative patients, respectively in the studied ward cubicle . Irrespe cubicle a, heavie00 hours a. 2) was approximately four times that in the MRSA-positive bed-units (from 1.5 to 2.0 cfu/cm2)  b,c. The p < 0.05) higher than that of the right-side handrail. However, with regard to the MRSA-positive bed-units, 54% of the MRSA isolates were recovered from the overbed rolling table surface as the dominant site of contamination. The mean MRSA concentration on the surface of the right-side handrail was found to be slightly higher  than that on the surface of the left-side handrail, while none was recovered from the bedside table specimens. With the focus on oxacillin resistance, sites of contamination were shown to be different among the bed-units of MRSA-positive and negative patients. The surface of the left-side handrail was found to be the most heavily contaminated site, containing 84% of all MRSA isolates recovered from the MRSA-negative bed-units. Indeed, the left-side handrail was found to have an MRSA concentration 11-fold significantly (p < 0.0001) from 4.4 ± 8.7 CFU/cm2 at 08:00 hours to 0.7 ± 0.26 cfu/cm2 at 12:00 hours due to the use of the QAC polymer  and bed-making could all contribute to the environmental dispersal of staphylococci. This is consistent with the results of this study, in which the overall surface staphylococcal burden from the sampled sites increased by 80% from 08:00 to 12:00 hours, when routine ward and clinical activities were maintained. Regarding the CNS strains that were recovered from all surfaces, the most clinically important CNS causing nosocomial infections is known to be Staphylococcal epidermidis, which is frequently sourced from the skin normal flora of patients and healthcare workers [Our results indicated that a large number (about 80%) of bedside surfaces was contaminated and that oxacillin resistance was prevalent, occurring not only around the beds of MRSA-positive patients but also of MRSA-negative patients, at one hour (08:00 hours) and five hours (12:00 hours) after a hypochlorite wiping (performed at 07:00 hours). The results reflected the natural contamination within the ward where hypochlorite cleaning was regarded as a routine practice, and authors are aware that it may have affected the staphylococcal identity and proportionate isolation. Surface MRSA contamination was commonly reported in bed-units or rooms occupied by MRSA infected or colonized patients ,13,14. W workers . NonetheHospital ward environments, especially high-touch surfaces, are rich reservoirs for the transmission of many microorganisms. The current results were consistent with those of previous studies reporting that handrails are the most frequently contaminated surfaces within a ward environment . Handraiin vitro cytotoxicity ranging from 99 to 100 percent has been tested on pathogenic microorganisms including Staphylococcus aureus, Treponena pallidum, Pseudomonas aeruginosa, Gonococcus, Colibacillus, Candida albicans, and SARS coronavirus [50) was determined as >10,000 mg/kg, which is essentially non-toxic. In particular it has no irritation to skin and eyes [The results clearly indicate that the application of JUC polymer on bedside surfaces effectively reduces both the incidence of staphylococcal contamination and bacterial concentration. The liquid preparation of JUC solidifies immediately when upon contact with the surface of skin or any fabric to form a two-sided film. The bonded film adheres firmly to the surface, and the positively charged film attracts the negatively charged cell walls and membranes of microorganisms to exert electrostatically destructive killing effects . So far,onavirus . The resonavirus . Toxicitand eyes . These fPseudomonas aeruginosa and multidrug resistant (MDR) gram-negative organisms including Stenotrophomonas maltophilia and vancomycin-resistant enterococci in the hospital environment. Such surface treatment should be abandoned if the MDR gram-negatives appear and cause problem for patients. To the best of our knowledge, this is the first study that has been conducted to evaluate the effectiveness of JUC spray as an enhanced surfactant disinfectant in addition to hypochlorite wiping in the general ward environment. The results were obvious on Staphylococci including the coagulase-negative and oxacillin-resistant strains, which warrants a large-scale investigation involving more hospital environments. Several researchers ,29 have The application of JUC OrganoSiQAC-based surfactant as a antimicrobial coating was found to be effective in reducing the incidence and bacterial concentrations of bedside staphylococcal contamination. It exerted long-lasting antimicrobial activity for at least four hours after application. The finding supports the application of JUC spray as a potential environmental decontamination strategy to prevent the transmission of clinically important pathogens in medical wards.


Several types of biodegradable polymer drug-eluting stents (BPDES) have been used for percutaneous transluminal angioplasty; however, the safety and efficiency of these BPDES have not been fully evaluated. A meta-analysis was, therefore, conducted to compare the clinical performance of BPDES with that of permanent polymer drug-eluting stents (PPDES) in unselected patients with coronary stenosis. PubMed, Web of Science, Medline and The Cochrane Library were searched for randomized clinical trials (RCTs) from January 2005 to January 2014. Trials that compared BPDES with PPDES in patients with coronary stenosis were considered. Twelve RCTs with a total of 15,938 patients with coronary stenosis were included in this meta-analysis. No significant difference was found between the two arms in the incidence of major adverse cardiac events (MACE) and definite or probable stent thrombosis (DpST) at the one-year follow-up (P>0.10). The use of BPDES, however, showed a tendency towards a lower risk of MACE (P=0.09) and a beneficial effect by reducing DpST episodes (P=0.04) at long-term follow-up, particularly when compared with the incidence of DpST at the one-year follow-up. BPDES also tended to be associated with a decreased late lumen loss in patients with coronary stenosis [instrumental variable =−0.04; 95% confidence interval =−0.08–0.00; P=0.05). In conclusion, the one-year outcomes following drug-eluting stent implantation showed BPDES were noninferior to PPDES in unselected patients with coronary stenosis. Long-term clinical outcomes, however, indicated that BPDES appeared to a present a lower risk of MACE and DpST. Over the past few decades a large number of patients have undergone percutaneous transluminal angioplasty (PTA), but they have also suffered a high risk of restenosis following PTA 30–50%) , which h0–50% , wTo be included in this meta-analysis, trials were required to meet the following criteria: i) Randomized clinical trials (RCTs) comparing a DES with a biodegradable polymer and a DES with a permanent polymer in patients undergoing percutaneous coronary intervention; ii) enrolment of >50 patients with available follow-up data for at least one of the clinical end-points or angiographic end-points at mid-term (≤9 months) and/or long-term (≤16 months). PubMed, Web of Science, Medline and The Cochrane Library were searched between January 2005 and January 2014 for RCTs on BPDES. The PubMed search strategy was formulated as follows: (‘biodegradable polymer’ OR ‘bioabsorbable’) AND (‘permanent’ OR ‘durable’) AND ‘clinical trials’. This search strategy was translated to the corresponding vocabulary of Medline, Web of Science and Cochrane Central Register of Controlled Trials. No language restriction was applied and the search was kept updated until January 2014.To select trials, the following steps were performed following trial identification by the main search: i) Exclusion of duplicates; ii) screening and selection of abstracts; iii) assessment for eligibility through full-text articles; iv) final inclusion. One author followed steps i) to ii) and another two authors followed steps iii) to iv) independently. Disagreements were resolved by discussion.Three authors independently assessed the risk of bias with the components recommended by the Cochrane Collaboration: Random sequence generation; allocation concealment; blinding of participants and personnel; blinding of outcome assessment; incomplete outcome data; selective reporting and other sources of bias . Trials Two authors independently performed data extraction on the trials. Any differences found were resolved by discussion. The information was collected regarding the main clinical characteristics  and angiographic characteristics . For both biodegradable polymer stent groups and permanent polymer stent groups, major adverse cardiovascular events (MACE), including cardiac mortality, MI or TVR were defined as the primary clinical outcomes. Other clinical or angiographic outcomes of interest included DpST, LLL in the stent and stenosis of lumen diameter (SLD) in the stent. In order to better compare the differences between short- and long-term follow-up results of BPDES and PPDES, the results are firstly classified into the one-year follow-up group (with a period of 12 months) and the long-term follow-up group (with a period of >12 months) according to the length of the follow-up period.2 and I2 statistics. Trivial heterogeneity was considered for P-values >0.1 or I2<50%, and a fixed-effect model would be used. A random-effect model replaced the fixed-effect model if P<0.1 or I2>50%, which suggested substantial and significant heterogeneity. The likelihood of publication bias was assessed graphically by generating a funnel plot for the primary end-points and angiographic outcomes. Subgroup analysis was performed based on the eluting drugs. Following the completion of data analysis, the GRADEPro system  was used for the scoring of the main analysis results to assess the value of each.Statistical analysis was performed using Review Manager . Summary statistics of dichotomous variables were presented as odds ratios (OR) and 95% confidence intervals (95% CI). Continuous variables were calculated as weighted mean difference with 95% CI. Following data pooling, statistical heterogeneity across trials was identified and evaluated by Cochrane Q χSeventy-two papers were identified from PubMed, 69 from Web of Science, 17 from The Cochrane Library and 54 from Medline. A total of 79 duplicates were excluded leaving 133 studies identified by the main search. A further 109 papers were excluded by reading the titles and abstracts so that 24 potentially relevant papers were identified. Finally, a total of 16 articles concerning 12 RCTs –26 with 2=4%), a fixed effect model was used . A funnel plot was used to assess the likelihood of publication bias, as presented in MACE data were acquired from 11 RCTs between nine and 16 months following the stent installation. As there was no significant heterogeneity  among these trials so a fixed effect model was selected. No significant difference was found between these two groups for DpST . The subgroup analysis of different eluting drugs showed comparable results and inter-group heterogeneity was low (I2=38%). The likelihood of publication bias is shown in DpST at the one-year time-point is reported in 2=0%) among these trials so a fixed effect model was selected . Results showed no significant difference between these two groups.MACE data were acquired from four RCTs regarding long-term follow-up . No hete2=0%). Subgroup analysis showed a statistically significant difference between the BP and PP groups at >36 months follow-up .Regarding long-term follow-up, the incidence of DpST in these two groups was similar but the BP group showed a tendency to reduced DpST compared with that in the PP group  among these trials and a randomized effect model was selected. The results  indicated that the difference between the BP and PP groups was considered to be statistically significant. Subgroup analysis showed that the biolimus-eluting stent (BES) was superior to the paclitaxel-eluting stent (PES) . A funnel plot of this result is shown in Regarding the 6–12-month follow-up, results of LLL in stent were acquired in eight trials, as shown in 2=70%, P=0.002) and a randomized effect model was used . Subgroup analysis indicated that BES could effectively decrease the severity of SLD in stent compared with PES  and sirolimus-eluting stent (SES) . This result indicated that a BES was more effective in reducing SLD in stent than other drug-eluting stents. Heterogeneity between these groups was significant (I2=94.6%). The likelihood of publication bias was assessed by funnel plot (SLD (in stent) is shown in nel plot .Following analysis of the data from RCTs by Review Manager, the GRADEPro system was used to evaluate the results. In conclusion, the quality of each result concerning long-term follow up was high. Details of the evaluation are shown in DES are a major breakthrough in the field of PTA, since they have markedly reduced the rate of acute stent thrombosis and the requirement for repeated revascularization procedures compared with bare-metal stents ,28. As tIn this analysis, the RCTs that were considered included numerous countries and regions; therefore, they covered the different races of the world. The analyzed BPDES group contained three types of stents with different eluting drugs: Sirolimus ,17,21,26The analysis of clinical events at the one-year follow-up showed that the incidence of clinical events due to BPDES was not significantly different from that due to PPDES. More accurately, the BPDES were noninferior to PPDES in safety profile one year following DES implantation. In the present study, the incidence of various clinical events in the analyzed RCTs was maintained at a low level, which provided evidence for the safety of modern stents.Nine RCTs reported angiographic outcomes. It was found that the degree of LLL in patients receiving BPDES was significantly lower than that in patients receiving PPDES. The Nobori I  and NoboBPDES were proposed to improve the long-term safety of DES as they was designed to reduce the incidence of LST and VLST. Recently, a long-term outcome of a pooled analysis of BPDES versus PPDES in patients with diabetes from three RCTs showed that BPDES were associated with comparable overall clinical outcomes at a four-year follow-up, and rates of DpST were significantly lower with BPDES . In the The main deficiencies in the present study are as follows: Firstly, the inclusion criteria did not specifically subdivide the lesion types of the patients in detail; therefore, the study lacked specific targets. As a result, the type of coronary stenosis the BPDES were more suitable for could not be determined; thus, it was not possible to provide a specific recommendation for clinical practice. Secondly, only four RCTs had a follow-up period >12 months and only two out of four RCTs had a follow-up period >36 months. The limited data did not make the analysis very reliable; the conclusion may be regarded as a reference with further research required to confirm the results.In conclusion, the results of the present meta-analysis suggest that BPDES were noninferior to PPDES in short-term results but superior to PPDES in long-term results. According to the clinical outcomes and angiographic outcomes at short-term follow-up, BPDES and PPDES exhibited no significant differences overall but the BES was superior to the PES. Subgroup analysis, however, demonstrated that the BES with a biodegradable polymer was superior to the PES with a permanent polymer. In this first meta-analysis comparing the data from long-term follow-up, the BPDES were found to exhibit an increased safety profile over time, particularly for incidences of LST and VLST in which the BPDES were superior to PPDES. This was consistent with the original purpose of BPDES design. Additional prolonged follow-up data, however, is required for an adequate comparison of the safety and efficacy between BPDES and PPDES to be conducted, in order to provide adequate and strong evidence for the selection of stents in clinical practice.