Obsessive-compulsive personality disorder (OCPD) traits and obsessive-compulsive disorder (OCD) are commonly associated with patients with Anorexia Nervosa (AN). The aim of this review was to systematically search the literature to examine whether OCPD and OCD are positively associated with excessive exercise in patients with AN.A systematic electronic search of the literature  was undertaken to identify relevant publications until May 2012.A total of ten studies met criteria for inclusion in the review. The design of the studies varied from cross-sectional to retrospective and quasi-experimental. Seven out of the ten studies reviewed demonstrated a positive relationship between OCPD and/or OCD in AN patients who exercise excessively, whilst three studies found a lack of relationship, or a negative relationship, between these constructs.There is evidence from the literature to suggest that there is a positive relationship between OCPD and excessive exercise in patients with AN. However, the relationship between OCD and excessive exercise is less clear and further research is required to qualify the strength of such relationships. Future research should utilise the most comprehensive and reliable clinical assessment tools, and address prognostic factors, treatment factors and specific interventions for patients with OCPD and/or OCD and excessive exercise. Anorexia Nervosa (AN) is recognised as one of the most serious chronic mental illnesses, with significant physical and psychosocial consequences . In ordeRothenberg  proposedExcessive exercise plays a detrimental role in the pathogenesis and maintenance of AN -21, and The reviewed research suggests that AN is associated with increased OCD symptomatology and a higher prevalence of OCPD traits. Research is warranted to determine personality and psychological variables for excessive exercise, in particular those that may be remedial to interventions . The aimThe search strategy was designed to identify all studies of patients with AN, in which OCPD or its traits, or OCD and its features were formally assessed, and in which excessive exercise was formally measured through clinical interview or clinical judgement.The following databases were systematically searched from April-beginning of May 2012: PsycINFO (1806-present), Medline (1950-present) and Web of Knowledge (1864-present). Reference lists from relevant articles were also manually searched for additional studies. The following search terms were used: (anorexia* OR anore* OR eating disorder*) AND  AND . Peer-reviewed research articles that focused on the relationship between exercise and obsessive compulsive disorder and/or obsessive compulsive personality traits in patients with anorexia nervosa were included. A total of 443 papers were retrieved from the electronic search. The titles and abstracts were screened to assess the suitability of papers. A second reviewer also screened a proportion of the titles and abstract to reduce selection bias. 79 papers were excluded from their title, and 302 papers were excluded from their abstract. The full text of 62 papers was read, and 54 were excluded. The reference lists of the final full text papers were searched manually, and a further two articles were retrieved. The second reviewer also read full texts of papers meeting the inclusion criteria, and there were no discrepancies in the inclusion of articles, thus a total of 10 studies were included in the review.A detailed map of the search strategy can be seen in Figure The final retrieved articles underwent quality assessment utilising an amended version of the original Quality Index by Downs and Black . The QuaA total of ten studies were reviewed: four studies utilised AN participants who were receiving inpatient treatment; three studies used inpatients and outpatients; one study used outpatients only, whilst another study stated that they recruited from four eating disorder services, but did not specify the settings. The final study included patients with AN who were from the multisite international Price Foundation Study of AN, BN and AN Trios studies, and their affected relatives who met lifetime diagnosis of AN  interview . These sOther assessment protocol for excessive exercise included questions regarding duration, frequency and intensity of exercise per week ,21,30 anOCD symptomatology was measured using different self-report questionnaires. The OBQ-44 assessed constructs of inflated responsibility/threat estimation, perfectionism/tolerance of uncertainty, and importance/control of thoughts . The ObsOCPD traits were measured through a number of methods. The EATATE interview  was inclp = .007) than non-excessive exercisers, unaffected by dietary status. Both excessive exercisers and non-excessive exercisers demonstrated a reduction in OCD symptoms between admission and discharge (p < .001). There was an interaction trend demonstrating that, after re-feeding, OC symptoms decreased less in the excessive exercisers group. It was also noted that patients who presented with excessive exercise reported a higher number of obsessive compulsive symptoms on the Maudsley Obsessive Compulsive Inventory (at admission and discharge) than a group of patients diagnosed with OCD [Davis and Kaptein  demonstrwith OCD .p < .01). Obsessive-compulsiveness was positively related to level of activity in AN patients (p < .01) and exercising participants also demonstrated higher OC symptomatology than non-exercisers (p = .02).Similar findings were demonstrated in studies by Davis et al. ,39 in whp < .01), obsessive-compulsive behaviours (p < .01), as well as higher eating disorder psychopathology (p < .01). Specifically, the Checking subscale from the OCI-R contributed uniquely and significantly to the overall model explaining weight control exercise, signifying that these obsessive beliefs and behaviours predict the variance in exercise for purpose of weight control, after controlling for eating disorder psychopathology. Furthermore, Shroff et al. [p < .001), and higher frequency of obsessions and compulsions (p < .001), when compared with AN patients who completed no or regular exercise.Naylor et al.  concludef et al.  reportedp = .038). Penas-Lledo et al. [p > .05). Finally, Holtkamp et al. [p = .705) and that obsessive-compulsiveness was not a significant contributor in the regression model predicting physical activity with other factors such as BMI, level of food restriction, depression and anxiety.However, Anderluh et al.  found noo et al.  found thp et al.  concludep = .03) than non excessive exercisers, and this was unaffected by dietary status.Davis and Kaptein  demonstrp < .05) and levels of self-oriented perfectionism (p < .05) than non-excessive exercisers. Anderluh et al. [p < .005) and cautious (p < .02), however they did not find any significant differences in current OCPD comorbidity (p > .05).Davis et al.  showed th et al.  reportedp < .05) and historically throughout the eating disorder (p < .01). Furthermore, Shroff et al. [p < .001), measuring factors such as concern over mistakes, personal standards, organisation and parental criticism.Davis and Claridge  demonstrf et al.  reportedThe aims of this systematic review were to critically examine evidence as to whether OCPD traits and/or OCD are associated with excessive exercise in patients with AN, and to determine the nature of relationships between these constructs. The results of the systematic review indicated a positive relationship between excessive exercise and obsessive-compulsive personality traits. However, the relationship between OCD and excessive exercise in AN patients is less clear with studies producing varying results.Davis et al.  proposedFurthermore, the findings of Naylor et al.  are consBewell-Weiss and Carter  reportedDavis et al.  speculatAnderluh et al.  found thThere are a number of limitations that were evident in the reviewed literature. The number of studies examined in the review (10) is small, and results must be interpreted with some caution as some of the studies did not support the association between OCD and excessive exercise. A number of the reviewed studies did not clearly differentiate between OCD and OCPD constructs in their presentation of results. Nine different types of measures were employed across the studies to assess OCPD traits and OCD symptomatology, many of these being self-report questionnaires, which may increase the incidence of socially desirable response styles. Others involved expert clinical assessment in the form of an interview . These Denial commonly occurs in anorexia nervosa, and subjective measures may have underestimated the amount of physical activity completed by patients. Such underestimation may have led to significant bias and consequently inaccuracy of data regarding the extent of exercise, and its relationships with coexisting OCD and/or OCPD. This potential room for bias has led to the utilisation of more objective means of assessing for physical activity in patients with AN, for example accelerometers ,78. Yet There are also a number of limitations in regards to participants utilised in the studies. The small sample size of Holtkamp et al.  affectedAs the vast majority of the studies employed a cross-sectional design, only relationships between variables could be determined, and there can be no demonstration of the direction of such associations. Acute starvation syndrome and severity of eating disorder psychopathology both have significant impact upon level of obsessive-compulsive symptoms and exercise . There wWhilst this review has focused on the relationships between OCPD traits and/or OCD symptomatology with excessive exercise in patients with AN, it would be remiss not to mention the significant relationships which excessive exercise shares with other psychopathologies, which have important implications for treatment. Studies included in the review demonstrated that AN patients who exercised excessively demonstrated lower minimum BMI and lower novelty seeking, but higher harm avoidance, persistence and cooperativeness . In otheFrom the research reviewed, it appears that there is a positive association between obsessive- compulsive personality traits in patients with AN who excessively exercise, yet the relationship with obsessive-compulsive disorder is less clear. Although it is known that excessive exercise is associated with poor treatment outcome in AN ,29 the eThe authors declare that they have no competing interests.SY undertook the systematic review and with PH prepared this manuscript for journal submission. PH, ST and PR assisted with the editing of the manuscript and data interpretation. All authors read and approved the final manuscript.SY submitted an alternate version of this manuscript as a requirement for the Doctor of Clinical Psychology/Master of Science thesis at University of Sydney. She is supervised by ST, PH and PR. PH, ST and PR are conducting a randomised controlled trial and evaluation of the Loughborough Eating Disorders Activity Programme (LEAP) referenced in this paper.


Based on previous research [μg/day dose of elemental chromium was unable to produce desired results [We read the research of Huang et al.  with greresearch , 4 and tresearch . Third,


Setaria tundra (Nematoda: Filarioidea) and an unidentified filarial species in mosquitoes in Germany” Parasites & Vectors 2012, 5:14.Comments concerning interpretation of the PCR xenomonitoring results in the article „Molecular detection of  Dear Editor,D. repens DNA detection in mosquitoes using the primers applied in the second round PCRs [et al. [et al. [D. repens sequences deposited in GenBank [In the context of an evaluation of the use of xenomonitoring (sensu ) for detund PCRs –5, was d [et al.  nor such [et al.  (to the  [et al.  of the p [et al. –5 perfec GenBank  – the pr GenBank , 5. We a GenBank .The absence of Dirofilaria spp. or other zoonotic filariae in our sample allows the conclusion that the risk of autochthonous infection in Germany is still very low, although dirofilariasis is emerging and spreading in Europe” [The authors interpreted every positive result of the screening real time PCR performed on DNA obtained from mosquito pools as positive result of filaria detection. However, the positive results of real time PCR might have been false positive; there were no positive PCR controls described, there were no negative PCR controls described. The sensitivity and the specificity of the PCRs applied in the study were not reported. Considering the above mentioned deficits, the following statement based on PCR results, seems unsupported: “ Europe” . The res Europe” . In the  Europe”  the auth Europe” . Only th Europe” . It woul


Assessment of volume and hydration status is far from easy and therefore technology such as bioelectrical impedance vector analysis (BIVA) may complement our examination techniques. This study highlights the fact that clinical assessment of volume balance and BIVA may correlate, but whether the routine use of BIVA will avoid significant volume overload in the critically ill remains unknown. Further studies are needed but at the moment appear a little way off. Critical Care, Jones et al. [2O) [In the current issue of s et al.  investigs et al. . Hydratis et al. . With des et al. . The golal. [2O) . TracersBioelectrical impedance analysis was originally introduced as a tool for assessing body composition and nutritional status, but early studies highlighted some limitations of this technique with variation in electrolyte levels, acute changes in hydration status and problems with some of the standard equations employed creating some disaffection , 7. BIVAIn the present study by Jones et al. , 344 meaIn summary, can BIVA guide fluid management in critically ill patients? As pointed out by Jones et al., this can only be addressed in well-designed interventional studies particularly with regard to the patient population. Given these preliminary results it seems unlikely that BIVA will play a major role in the critically ill level 3 patient with sepsis where rapid fluid shifts are occurring or in the unstable postoperative patient, but the technique may inform in less acute situations such as renal replacement therapy in a step-down unit.


Gertig and Hanisch , always on the edge of causing massive destruction when provoked. Nevertheless, microglia are responsible and/or strongly contribute to the maintenance of CNS homeostasis, immune surveillance and synaptic plasticity. Until now, it was believed these complex processes depended on two polarizing stimuli received by microglia: the “bad” ones leading to a classical pro-inflammatory response (M1), and the “good” ones leading to a typical anti-inflammatory profile (M2). However, under M1 and M2 polarizing conditions, microglia can  Hanisch  extensiv Hanisch  distingu Hanisch  show tha Hanisch  extensiv Hanisch . The groa et al.  review ca et al.  propose l et al.  show howo et al.  comment 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.


Venous thromboembolism (VTE) is a common complication of malignancies and epidemiological studies suggest that lung cancer belonged to the group of malignancies with the highest incidence rates of VTE. Risk factors for VTE in lung cancer patients are adenocarcinoma, NSCLC in comparison with SCLC, advanced disease, pneumonectomy, chemotherapy including antiangiogenic therapy. Other risk factors are pretreatment platelet counts and increased release of TF-positive microparticles. Elevated D-dimer levels do not necessarily indicate an increased risk of VTE but have been shown to be predictive for a worse clinical outcome in lung cancer patients. Mechanisms responsible for the increase in venous thrombosis in patients with lung cancer are not understood.Currently no biomarker is recognized as a predictor for VTE in lung cancer patients.Although several clinical trials have reported the efficacy of antithrombotic prophylaxis in patients with lung cancer who are receiving chemotherapy, further trials are needed to assess the clinical benefit since these patients are at an increased risk of developing a thromboembolism. Thromboembolism is a well recognized complication of malignant disease and it is known that cancer patients have a higher incidence of venous thromboembolism (VTE), including pulmonary embolism (PE) and deep venous thrombosis (DVT), compared to the general population .The incidence of thromboembolic disease in general population is relatively low, about 1–3 per 1.000 per year  while amIt is estimated that cancer is responsible for 20 % of all cases of incident VTE .Epidemiological studies suggest that lung cancer belonged to the group of malignancies with the highest incidence rates –9 of VTEThe association between VTE and lung cancer has been reported more than 20 years ago , 11.Lung cancer is the leading cause of cancer death in the United States for both men and women, with over 156,000 deaths in 2011 in the United States . It is aIn recent years, there has been increasing attention to the phenomenon of VTE in lung cancer patients and several studies have been made to better characterize it.It is recognized that patients with lung cancer are at risk for development of VTE. The incidence of VTE in lung cancer patients has been described in several studies.et al. [Chew et al.  investgaet al. .In a retrsospective analysis of a lung cancer cohort of 6732 patienst , VTE occurred in 13,9 % of the lung cancer cohort and 1,4 % of the control cohort .In another retrospective study of 1940 patients with diagnosis of lung cancer, thromboembolic events were documented in 9.8 % cases, venous thromboembolic complications in 78 %, arterial thromboembolic complications in 27 % cases. About venous thromboembolic complications, it was documented deep venous thrombosis in 55 % cases and pulmonary embolism in 66 % cases .et al. [Recently Zhang et al.  describeet al. .et al. [The incidence of thrombotic event was evaluated in a group of 950 patients with lung cancer by Kadlec et al. . 91 . Almost all of these unexpected VTE cases were significantly associated with a diagnosis of metastatic-stage cancer [White et al.  identifie cancer .In a more recent study, it was observed among 207 subjects with lung cancer that one-third of VTE events was incidentally discovered .et al. [Calleias et al.  have evaet al. . Patientet al. .Mechanisms responsible for the increase in venous thrombosis in patients with cancer are not understood. Rather than one unifying mechanism, the etiology is likely multifactorial with different factors assuming lesser or greater degrees of importance depending on the clinical setting.Over the years, researchers have investigated intrinsic properties of tumor cells that lead to a prothrombotic state. It is recognized that tumors shed membrane particles that contain procoagulant activity including tissue factor and membrane lipids that propogate the coagulation response , 24.Several studies have focused on the role of Tissue Factor (TF) in the pathophysiology of cancer associated thrombosis.Tissue factor (TF) is the physiologic initiator of coagulation and is commonly expressed in a variety of malignancies , 26. OveIn the past, some studies demonstrated the expression of TF by lung cancer cells , 29.et al. [et al. [Sawada et al.  have pre [et al.  also demMore recently the expression TF protein and/or mRNA have been documented in malignant cells or tissue in NSCLC .et al. [Sato et al.  describeet al. .et al. [Similarly, Del Conde et al.  reportedet al. .Cancer increases the risk of thromboembolic events. Risk factors, such as age, gender, bed-rest, venous catheters, surgery, chemotherapy with or without adjuvant hormone therapy, radiotherapy and infections, also increase the risk of thrombosis in cancer patients.Rates of VTE vary substantially between cancer patients, considerably depending on clinical factors, the most important being tumor type and stage.In this review we have focused on studies which investigated factors could be identified as predictors of vascular events in lung cancer and their prognostic significance.Thrombocytosis can result in thrombosis and is frequently observed in patients with malignancies The mechanism underlying development of thrombocytosis in lung cancer patients remains unclear; one of possible mechanisms might be tumor-associated elevation of bone marrow-stimulating cytokines such as interleukin (IL)-6, IL-1 and macrophage colony stimulating factor (M-CSF). These cytokines could exhibit thrombocytosis. Incidence and prognostic significance of thrombocytosis in patients with lung cancer has been investgated in some studies .Pedersen and Milman  examinedet al. [p < 0.0001). Therefore preoperative platelet count was a prognostic factor for resectable NSCLC patients [Tomita et al.  evaluatepatients .p < 0,001) [These findings are supported by a more recent study  that ass< 0,001) .et al. [9 was associated with OR for major thromboembolic events was 3.66 (2.25–5.96) — a statistically significant value. Thrombocytosis was found in patients with advance disease, more commonly in men, with no apparent correlation with the clinical stage of the disease [Recently Kadlec et al.  observed disease .et al. in a restrospective study of consecutive 281 patients with lung cancer did not observe a significant correlation between vascular events and thrombocytosis [On the contrary, Demirci ocytosis .Plasma levels of D-dimer are elevated in cancer patients. Activation of the extrinsic coagulation system and the fibrinolytic cascade within a tumour is thought to be related with growth, invasion and metastasis. Although high D-dimer was described as a predictor of VTE in earlier works, it is now recognized that it does not necessarily indicate an increased risk of VTE in cancer patients .Several studies have demonstrated that elevated D-dimer levels are associated with a poor prognosis in cancer patients. In patients with both non-small cell and small cell lung cancer, increased plasma D-dimer levels were shown to be predictive for a worse clinical outcome .In addition in cancer lung patients it has been shown that D-Dimer plasma levels decrease or increase after response and progressive disease, respectively, and can act as a predictive factor of the evolution of the disease .et al. [p = 0.0001) [Recently Ferroni et al.  hypotheset al. . Overall 0.0001) . The use 0.0001) .With regard to other hemostatic factors, there are different results in the literature.et al. [Gabazza et al.  found siet al. [In a prospective study Zecchina et al.  investigElevated levels of procoagulant factors, such as lupus anticoagulant, anticardiolipin antibodies to factor VIII, and certain cytokines such as interleukin-6 and tumor necrosis factor, have also been identified as increased risk factors for VTE in patients with lung cancer , 29.et al. in a study population of 950 lung cancer patients did not find significantly higher medians of coagulation parameters  [Recently Kadlec nd aPTT) .Over the years several studies investigated the VTE risk in different hystological types of lung cancer.et al. [In the past, autopsy and retrospective studies indicated that various adenocarcinomas are most strongly associated with VTE , 46. Thiet al.  investiget al. .In a restrospective analisys of a large cohort of patients with NSCLC and SCLC, was observed that adenocarcinoma histology  and advancing cancer stage  were significant predictors of developing VTE within 1 years of NSCLC diagnosis .et al. and Lee [Recently Kadlec  and Lee , 48 idenet al. [Tagalakis et al.  describeet al. [In a retrospective analysis of lung cancer patients, Alexander et al.  observedet al. .et al. [Differently not a significant correlation between vascular events and histological type, or TNM stage was observed in a retrospective study by Demirci et al. .VTE was a significant predictor of death within 2 years for both NSCLC and SCLC, HR = 2.3, 95 % CI = 2.2–2.4, and HR = 1.5, 95 % CI = 1.3–1.7, respectively .Cancer therapy itself has been shown to increase the risk of VTE, whether it be chemotherapy, antiangiogenic therapy, or hormonal therapy.Combined chemotherapy (in combination with radiotherapy) is the current standard treatment for advanced stage NSCLC as well as in the treatment of SCLC patients and VTE is a well known complication of anticancer therapy , 50.How chemiotherapy contributes to VTE risk has not to our knowledge been elucidated completely but likely involves a combination of direct endothelial damage and down regolation of endogenous anticoagulants .et al. [In an observational study by Blom et al. , the oveet al. .et al. [Khorana et al.  analysedet al. .et al. [The results of this study are supported by Hicks et al.  that havet al. .et al. [Numico et al.  prospectet al. .Recently a retrospective study of lung cancer patients showed that chemotherapy-treated patients experienced thromboembolism more often than patients who did not receive chemotherapy (HR 5.7 95 % CI 2.2–14.8) .Among lung cancer patients receiving chemotherapy, it was described that the majority of VTE events occurring within 6 months of initiation of chemotherapy . The preWith regard to antiangiogenic chemotherapy, researchers have focused on neoangiogenesis inhibition with VEGF inhibitor bevacizumab.While some trials suggested that bevacizumab may be associated with increased risk for VTE , others et al. [p = 0.031) but was not found to be associated with an increased risk of venous thromboembolic events [Scappaticci et al.  conductec events .et al. [p < 0.001) compared with controls [These data are in contrast to a recent systematic review and meta analysis by Nalluri et al. ; resultscontrols .Antiangiogenic agents also contribute to thrombosis, perhaps through endothelial cell and platelet activation .Postoperative use of angiogenesis drugs EGFR-TKI application were high risk factors for VTE in lung cancer patients .With regard to metalloproteinase inhibitor prinomastat there are divergent data.et al. [According to Behrendt et al. , the useet al. .In contrast to these data in a more recent study the addition of another metalloproteinase inhibitor BMS-275291 to chemotherapy in patients with advanced NSCLC did not increase the risk .The addition of aprinocarsen, a protein kinase C- alfa antisense oligonucleotide, to standard chemotherapy in patients with advanced stage NSCLC, significantly increased thromboembolism .It is known that patients undergoing surgery for cancer have a higher risk of postoperative DVT, despite thrombosis prophylaxis , than thAmong patients who undergo surgical resection due to oncological processes, VTE is considered a major cause of mortality and may serve as an important predictor of survival.et al. [With regard to lung cancer, Ziomek et al.  analyzedet al. .et al. [Lyman et al.  establisPneumonectomy has been associated with a higher risk of VTE than lobectomy for stage I and II lung cancer, due to greater activation of coagulation, which appears from the seventh postoperative day .et al. [Mason et al.  describeet al. [Yang et al.  exploredet al. .et al. [Recently Kadlec et al.  reportedet al. [Differently Dentali et al.  have obset al. .et al. [et al., this low prevalence may be attributable to the use of thromboprophylaxis protocols, which include the use of anticoagulant drugs and mechanical measures (intermittent compression systems and elastic stockings) and early ambulation [Gomez-Hernandez et al.  have fouet al. . Accordibulation .et al. [A recent published review by Christensen et al.  analysedet al. .et al. [With respect to mortality, it was described that pulmonary embolism is the second cause of mortality after pneumonectomy for a malignancy and such patients have the highest risk of dying from pulmonary embolism . Weder eet al.  in theiret al. .Interestingly, mortality due to VTE has been reported to occur predominantly in patients with squamous cell lung carcinoma following surgery .In cancer patients the risk of VTE is particularly high in association with surgery for cancer and chemotherapy.et al. [P = .032) without increased bleeding [Haas et al.  investigbleeding .et al. [p = 0.04) and thromboembolic events . Furthermore anticoagulation showed significant improvement in survival at 1 year  and at 2 years , especially for patients with SCLC and prolonged life expectancy [Zhang et al.  have recpectancy . Betweenpectancy .In a retrospective analysis of lung cancer patients, patients receiving chemotherapy experienced thromboembolism more often than patients who did not receive chemotherapy (HR 5.7 95 % CI 2.2–14.8) . PharmacRecently the neutrophil-lymphocyte ratio (NLR) was identified as a potentially useful marker for predicting clinical outcome in patients with anticoagulation for VTE. The study has demonstrated that NLR at the time of VTE diagnosis could be a useful biomarker for predicting the response and prognosis following anticoagulation in patients with lung cancer and VTE .The CARMEN study is an interesting french observational study, in which the researchers evaluated the adhesion to guidelines for treatment of VTE in hospitalized patients .Among cancer patients, 64 % had metastatic disease. Cancer sites were gastro-intestinal (25 %), gynecologic (23 %), pulmonary (21 %), hematological (14 %), urologic (10 %), or other (8 %) .It was observed that overall adhesion to guidelines was present in 59 % of patients. During initial treatment, adhesion was high but dropped during the long-term mantenance. Lung and hematological malignancies were significantly associated with the highest and lowest rates of adhesion .et al. [Conversely Alexander et al.  made a set al. .Venous thromboembolism, including pulmonary embolism and deep venous thrombosis, is a common occurrence within the cancer population. Lung cancer is one of the most common cancer in western countries with reported high incidence of VTE. VTE in hospitalized lung cancer patients is associated with longer length of stay, higher inpatient mortality rates, increased cost and greater disability upon discharge compared to other inpatient lung cancer patients . Risk faCurrently no biomarker is recognized as a predictor for VTE in lung cancer patients.Although several clinical trials have reported the efficacy of antithrombotic prophylaxis in patients with lung cancer who are receiving chemotherapy, further trials are needed to assess the clinical benefit since these patients are at an increased risk of developing a thromboembolism.


Life devoted to The Origins and Early Evolution of RNA, 17 papers explore a remarkably broad range of topics surrounding this hypothesis. I would not go so far as to say the hypothesis is experiencing a mid-life crisis. However it is clear that it has generated a spectrum of viewpoints, from ardent devotees to outright skeptics. I perceive all these vantage points as giving us a richer appreciation of the chemical origins of life.The RNA World is now some four billion years behind us, but only recently turned 50 as a human hypothesis. As early as 1962 Alex Rich suggested that RNA might have a phenotype in addition to its informational role [i.e., RNA enzymes, and thus operating on reversible reactions) generate such products. Martin et al. also briefly consider the role of 2´,3´-cyclic phosphates in their paper, which is an in-depth survey of the problems and potentials of RNA ligase, nucleotide synthase, and RNA replicase ribozymes [The Issue was initiated by a cogent argument from Scott and colleagues that the 2´,3´-cyclic phosphate version of nucleotides should be considered a viable prebiotic source of monomers for the abiotic polymerization into RNA [et al. revisit the role of formamide as both a reactant and a solvent for activated nucleotide synthesis that obviates much of the water problem [The key problem of how RNA nucleotides, if present prebiotically, could polymerize into oligomers in the face of an uphill battle against hydrolysis in an aqueous medium, was taken up by two papers in the Special Issue. Hashizume  details  problem . Of note problem ,7.The transition from “short-mers” to “longer-mers” was also investigated by Mungi and Rajamani  and KanaOnce RNAs were long enough, the RNA World hypothesis generally turns to how the evolutionary process could speed up. Kun and Szathmáry, utilizing a recent influx of experimental data on RNA function, explore the nature of RNA fitness landscapes . A very et al. [et al. [e.g., [Another “next step” in RNA World discussions is the origin and evolution of the genetic code. This problem was pondered long ago by Jukes , Woese , and othet al.  take thiet al.  takes th [et al.  also tak [et al.  also con. [e.g., , and woret al. [Once the RNA World had a foothold, a next logical question is, how did life bring in DNA? In today’s biology, DNA synthesis requires a very challenging enzymatic reaction catalyzed by the ribonucleotide reductase (RNR) protein enzymes, which use RNA as a substrate. Given the complexity, yet apparent antiquity, of this reaction, the evolutionary provenance of these enzymes is of extreme importance. Lundin et al.  provide i.e., RNA per se). The same stance is taken by van der Gulik and Speijer, who argue forcefully on catalytic grounds that an RNA world without amino acids could never have existed [et al. [The last group of papers in the Issue deal with the highly probable case where the RNA World was not strictly consisting of RNA, and that peptides or other molecules either initiated life or co-evolved with RNA. The so-called “RNP World” – for ribonuleoprotein world – posits that RNA could not have had enough catalytic prowess without the help of peptide cofactors. This is a rather attractive hypothesis and various authors have varying degrees of opinions on its strength. The “Strong RNP World”  is advo existed . Smith e [et al. , perhaps [et al.  makes pe [et al. .In sum, all of these papers serve to richen the discussion of the RNA World, in all of its various forms. Although definitive confirmation of any of these ideas may require a time machine, I sense that we are at the precipice of a unified theory that accommodates a wide spectrum of RNA-related observations.


Lack of clean water sources, starvation, insufficient hygiene, and poverty are some of the greatest barriers to health for the world's growing population. All these features are closely associated with parasitic infections. Approximately one third of the world's population has been infected with parasites at some point in their lives and these infections are often life-threatening. Since most parasitic diseases progress with few to no symptoms, patients do not obtain accurate diagnosis and treatment in a timely manner. Further, there are currently no accessible and effective antiparasitic vaccines despite enormous efforts and monetary investment into the development of vaccines, drugs, and treatments to combat these infections. Today parasitologists are looking for new alternatives for treatment such as immunotherapies, gene manipulation, or transfection in order to improve their fight against these “elusive” organisms. It is also clear that more studies on various parasites are necessary, even those which have low incidence, so that we are well-prepared during threats of reemerging parasitic infections.In this special issue, we bring together several reviews as well as original reports that are intended to provide a summary of some of the current knowledge regarding the “immunology and cell biology of parasitic diseases.” These papers include basic, clinical, and epidemiologic studies that we believe are interesting and very important in our field. The first section of this special issue is focused on helminthic diseases ranging from vaccine development to helminth therapy and includes some research on the basic mechanisms of susceptibility, modulation, and protection against these parasites. Schistosoma mansoni Antigen in Cutaneous Leishmaniasis Patients,” describes how helminth-derived molecules from S. mansoni can modulate dendritic cell activities during Leishmania infection. Additionally, V. H. Salazar-Castañon et al. have written an interesting review on how different helminth infections can improve or worsen the development of malaria in “Helminth Parasites Alter Protection against Plasmodium Infection.” Data from their epidemiological and experimental studies indicates that helminth infections are a double-edged sword, in the context of malaria.The work by D. M. Lopes et al., “Dendritic Cell Profile Induced by T. solium in their original paper “Characterization of a Thioredoxin-1 Gene from Taenia solium and its Encoding Product.” Thioredoxin-1 is an essential component of the thioredoxin system and it performs functions such as antioxidative, protein-reducing, and signal-transducing ones for development, proliferation, migration, apoptosis, inflammation, and metabolism. It is secreted by T. solium and is able to modify the immune response by driving a Th2 biased response and allowing for the establishment of this parasite. Further, E.-V. Marcela et al. studied how the metacestodes of T. crassiceps “communicate” when they grow in vitro in a crowded manner. Their paper is entitled “Crosstalk among Taenia crassiceps (ORF Strain) Cyst Regulates Their Rates of Budding by Ways of Soluble and Contact Signals Exchanged between Them.”The next series of papers are focused on the biology of helminths. L. Jiménez et al. characterize the thioredoxin-1 gene and gene product from Taenia crassiceps without Modifications of Parasite Loads.” Using the same model, M. Khumbatta et al. show the important role that somatostatin has on the immune response and susceptibility to experimental cysticercosis in their paper “Somatostatin Negatively Regulates Parasite Burden and Granulomatous Responses in Cysticercosis.” A couple more papers focused on the immune response elicited by different helminth parasites. A. Prasad et al. report the first advances on immune response to the flat worm Paramphistomum epiclitum in their contribution “Evaluation of Antibody Response to Various Developmental Stage Specific Somatic Antigens of Paramphistomum epiclitum in Goats.” Y. Gu et al. report a promising experimental vaccine against trichinellosis in their paper “Protective Effect of a Prime-Boost Strategy with the Ts87 Vaccine against Trichinella spiralis Infection in Mice.” We have another paper on immunoregulation by helminths in the contribution of Y. Ledesma-Soto et al., “Extraintestinal Helminth Infection Limits Pathology and Proinflammatory Cytokine Expression during DSS-Induced Ulcerative Colitis: A Role for Alternatively Activated Macrophages and Prostaglandins.” This paper suggests that helminth infections can modulate intense inflammatory processes such as colitis through different mechanisms and ameliorate signs of illness in mice.Next, we put together a series of papers related to the development and regulation of immune responses to different helminths. K. E. Nava-Castro et al. demonstrated how early exposure to estrogens can imprint the immune response and have a positive or negative outcome during adulthood. Their results can be found in the paper “Diethylstilbestrol Exposure in Neonatal Mice Induces Changes in the Adulthood in the Immune Response toFinally in this helminth section, B. Moguel et al. show in their review “Transfection of Platyhelminthes” how knowledge of the genome of helminths and genetic manipulation can be useful for designing new and more effective antihelminthic drugs. They also explain the molecular crosstalk that occurs between the host and parasite which has been partly inaccessible to experimentation.P = 0.049) and homozygous mutant  were significantly associated with amoebic liver abscess when compared with homozygous wild type (QQ).The second section of this special issue deals with protozoan infections which are widely spread around the world. We started this section with an old “friend” from developing countries, the amoeba, which causes amebiasis and remains as a public health problem in this part of the world. A. Aceves-Cano et al. present an original research “Morphological Findings in Trophozoites during Amoebic Abscess Development in Misoprostol-Treated BALB/c Mice,” where they show how trophozoites are altered with this kind of treatment. At human level A. K. Verma et al. contributed with the paper “The Trend in Distribution of Q223R Mutation of Leptin Receptor Gene in Amoebic Liver Abscess Patients from North India: A Prospective Study,” they show that heterozygous mutant  and E. dispar (a noninvasive strain). Talamás-Lara et al. demonstrated a dramatic difference in the ability to phagocyte in these two species in their paper, “Erythrophagocytosis in Entamoeba histolytica and Entamoeba dispar: A Comparative Study.” They discovered that E. histolytica displayed a superior erythrophagocytosis activity which possibly contributes to its more pathogenic nature.D. M. Meneses-Ruiz et al. developed a new experimental vaccine focused on the expression of a lectin in their paper “Protection against Amoebic Liver Abscess in Hamster by Intramuscular Immunization with an Trypanosoma, Leishmania, and Plasmodium.Other protozoan parasites included in this special issue are Trypanosoma cruzi Reduce Cross-Reaction with Leishmania spp. in Serological Diagnosis Tests.” The aim of this study was to improve serological tests already standardized for Chagas disease diagnosis, by using a high molecular weight protein fraction from T. cruzi extracts. They developed an easier and cheaper assay which is much needed in poor countries that suffer from this debilitating disease. From the same group, A. Vizcaíno-Castillo et al. describe the processes of acute inflammation derived after experimental T. cruzi infection in their original paper “Exacerbated Skeletal Muscle Inflammation and Calcification in the Acute Phase of Infection by Mexican Trypanosoma cruzi DTUI Strain.” In Leishmaniasis research, we have several interesting papers, such as “CK2 Secreted by Leishmania braziliensis Mediates Macrophage Association Invasion: A Comparative Study between Virulent and Avirulent Promastigotes” by A. M. B. Zylbersztejn et al., where the authors comparatively analyze the effect of the kinase enzyme CK2 of virulent and avirulent L. braziliensis strains on parasite growth and macrophage invasion. They show interesting data that demonstrates that CK2 has a critical influence as a mechanism of invasion used by L. braziliensis. On the clinical side, N. Verma et al. show important cases where immunological changes were observed according to the treatment carried out by the patients, in their work “Clinicopathological and Immunological Changes in Indian Post Kala-Azar Dermal Leishmaniasis (PKDL) Cases in relation to Treatment: A Retrospective Study.” In another review by S. A. G. Da-Silva et al. entitled “The Dialogue of the Host-Parasite Relationship: Leishmania spp. and Trypanosoma cruzi Infection,” they have analyzed the host-parasite interaction in both Leishmania spp. and Trypanosoma cruzi infections. This review is comprehensive and is interesting since it indicates that there are some differences in host invasion strategies between these two major protozoan parasites. Three schematic figures summarizing the main escape mechanisms of Trypanosoma and Leishmania and modulation of host cells are included.A. Y. Cervantes-Landín et al. contributed their work “High Molecular Weight Proteins of P. berghei ANKA,” where the authors highlight the important effect of sexual hormones on the development of Plasmodium infection.Another interesting original study related to malaria is presented by N. A. Mosqueda-Romo et al. in “Gonadal Steroids Negatively Modulate Oxidative Stress in CBA/Ca Female Mice Infected withFinally, a review of the basic cell biology of protozoan parasites and their virulence is discussed by C. Muñoz et al., in their latest paper, “Role of the Ubiquitin-Proteasome Systems in the Biology and Virulence of Protozoan Parasites.”We believe that this compilation of original research as well as the latest reviews written by authors from all around the world is a small sample about interesting yet complicated field of immunoparasitology. This research shows us that it is necessary to develop a deeper knowledge about the different mechanisms that parasites employ to invade the host and to avoid antiparasitic immune responses. We also require a better understanding of how they develop resistance to constant exposure to old drugs.Luis I. TerrazasAbhay R. SatoskarMiriam Rodriguez-SosaAbraham Landa-Piedra


Sapajus spp). Specifically, variation in the facial width-to-height ratio (fWHR) was positively correlated with alpha status and a composite measure of assertiveness. This novel finding adds to a growing body of evidence indicating that variation in facial structure reliably maps onto individual differences in dominance-related phenotypes.A recent paper by Lefevre et al.  in PLoS Research into fWHR was propelled by an anthropological study of human skulls indicating that fWHR was a size-independent sexually dimorphic feature of the human skull that arose around puberty coincident with the rise in pubertal testosterone  with aggression , a careful examination of Figure 4 from Lefevre et al. (r(23) = 0.54, p = 0.005], but not alpha monkeys . I decided to perform a re-analysis of Lefevre and colleagues' data which were freely available on the PLoS One website  to investigate the extent to which the link between fWHR and assertiveness was driven by low status monkeys. In this model, assertiveness was the dependent variable and I included fWHR and alpha status on Step 1 and the fWHR-×-alpha status interaction on Step 2. As per Lefevre et al. .In their paper, Lefevre et al.  reportede et al.  certainle et al. , I also B = −1.32, SE = 0.30, p < 0.01) and fWHR  and a trend for a fWHR-×-alpha status interaction . Lefevre et al. (p < 0.05). However, the lack of statistical significance is almost certainly due to a lack of statistical power given the small sample size (n = 43). Because previous work has found that the relationship between fWHR and aggressive behavior is specific to individuals with relatively low social status  but not alpha monkeys  , sex-×-fWHR (p = 0.84), or sex-×-alpha status-×-fWHR interactions (p = 0.15), suggesting that the relationships between fWHR, alpha status, and assertiveness were not moderated by sex.Results revealed main effects of alpha status . Thus, future work will require a larger sample to verify the extent to which fWHR maps onto dominance-related traits, whether such effects are moderated by social status, and whether relationships between fWHR, social status, and assertiveness hold across males and females.One important finding from Lefevre et al.  and the In summary, these findings in brown capuchin monkeys, along with work in humans (Goetz et al., The author 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.


Glycation is important in the development of complications of diabetes mellitus and may have a central role in the well-described glycaemic memory effect in developing these complications. Skin fluorescence has emerged over the last decade as a non-invasive method for assessing accumulation of advanced glycation endproducts. Skin fluorescence is independently related to micro- and macrovascular complications in both type 1 and type 2 diabetes mellitus and is associated with mortality in type 2 diabetes. The relation between skin fluorescence and cardiovascular disease also extends to other conditions with increased tissue AGE levels, such as renal failure. Besides cardiovascular complications, skin fluorescence has been associated, more recently, with other prevalent conditions in diabetes, such as brain atrophy and depression. Furthermore, skin fluorescence is related to past long-term glycaemic control and clinical markers of cardiovascular disease. This review will discuss the technique of skin fluorescence, its validation as a marker of tissue AGE accumulation, and its use as a clinical tool for the prediction of long-term complications in diabetes mellitus. Worldwide, the prevalence of diabetes mellitus is increasing at an alarming rate, mainly because of an increasing incidence of type 2 diabetes mellitus (T2DM) . The conAGEs are formed in a multistep process by glycation and oxidation of free amino groups in proteins, lipids, and nucleic acids. These AGEs promote tissue dysfunction through cross-linking of long-lived molecules and through binding to the receptor for advanced glycation endproducts (RAGE) . AGEs acet al. [In 1986, Monnier et al.  first pret al. .However, assessment of AGEs in skin biopsy is not suitable for clinical use, because of the invasive method and high costs. In 2004, a first report appeared that skin fluorescence, non-invasively assessed, was related to AGE levels in dermal biopsies in diabetes patients and in healthy controls . The derSo far, two devices have been used for assessment of skin fluorescence in clinical studies: the AGE Reader  and the SCOUT DS SF spectrometer . Although both devices have been developed to reflect skin AGE accumulation, the measurement techniques differ, and the measurement values are not directly comparable. Therefore, skin fluorescence as measured by the AGE Reader, previously called Autofluorescence Reader (AFR), will be referred to as skin autofluorescence (SAF). Skin fluorescence as measured by the SCOUT device will be referred to as skin intrinsic fluorescence (SIF). When general statements on either device are made, the term ‘skin fluorescence’ will still be used.SAF is measured on the volar side of the forearm. Care should be taken to perform this measurement in an area with normal skin with minimal sunlight exposure. The AGE Reader contains a UV-A light emitting lamp that emits light with a peak wavelength of 360–370 nm. Light reflected and emitted in the 300–600 nm range from the skin is measured in the research version by an inbuilt spectrometer, using a UV glass fibre. In later, simpler versions, the spectrometer is replaced by a set of photodiodes with peak sensitivities for different wavelengths. Before every measurement, dark and white reference readings are performed to correct for background light and to calculate reflectance, respectively. Initially, SAF measurements were not considered for analysis if the reflectance level was less than 10 %. After introduction of more sophisticated and validated skin colour correction software (version 2.3), this limit was lowered to 8 %. This adaptation allowed the use of SAF in a broader group of persons with dark skin colour. To correct for differences in light absorption, SAF is calculated as a ratio of excitation light (300–420 nm) to emitted light (420–600 nm). Consequently, SAF is expressed in arbitrary units (AU). Intra-observer variation of repeated autofluorescence measurements is 5 % to 6 % within one day in different publications [Although the exact molecular structures and the diversity of species contributing to skin fluorescence are not established, tissue fluorescent species have their own specific excitation-emission spectrum. The used wavelength band of the AGE Reader was selected such that the major contribution in fluorescence comes from fluorescent AGEs. The majority of identified AGEs are characterized by fluorescence in the area around an excitation wavelength of 370 nm and an emission of 440 nm –11. For SCOUT measurements are performed on the inner side of the forearm and are corrected for factors that affect light scattering and absorption as well. SIF is excited with a light-emitting diode (LED) centred at 375 nm, and the emission is detected over 435–655 nm  . In secoSeveral validation studies have been conducted to determine the performance of SAF as a tool for measuring dermal AGE accumulation. First, SAF was compared with several specific fluorescent and non-fluorescent AGEs in skin biopsies of diabetes patients and control subjects. Later, SAF was validated in a comparable way in renal failure and some other conditions. Moreover, SAF was compared to another classical AGE assay method for measuring AGEs, namely collagen-linked fluorescence , 15, 16.versus control subjects. This study demonstrated that SAGE, a skin derived skin fluorescence parameter, could accurately classify disease in a case-control population [SIF was originally based on a comparison of non-invasive fluorescence spectroscopy and skin collagen AGEs (determined using chromatographic and mass spectrometric methods) in a pig skin collagen model . Furtherpulation . HoweverA remark on the use of skin fluorescence deserves attention; skin fluorescence measurements may be influenced by dark pigmentation, the use of skin products, the fasting state, and lifestyle factors.For the AGE Reader, it has been demonstrated that SAF levels show expected relations with calendar age and presence of diabetes up to Fitzpatrick  skin phoSeveral skin products may affect the measurement of SAF, especially sunscreens and skin tanners. However, some conventional skin creams may also make the measurement unreliable. When possible, persons should be asked to avoid use of skin products several days before a SAF measurement. Otherwise, persons should be asked about recent use of skin products .et al. [An increase in SAF was seen 2 h postprandial in a study performed by Stirban et al. . This maet al. . TherefoN-acetyltransferase 2 polymorphism, (pack-years of) smoking, and coffee consumption. In the T2DM population, 47 % of the variance in SAF was explained by the same factors except for coffee consumption.As AGEs accumulate over time, SAF increases with aging . In addiversus post-glucose challenge) was comparable to the variation (5.5 %) for two non-fasting measurements [For the SCOUT device, it was claimed that the skin colour range, in which reliable measurements could be made, was somewhat broader than for the AGE Reader. In subjects at risk for T2DM, it is observed that SIF measurements are not majorly influenced by the fasting status since the variation (5.7 %) for fasting subjects  studies showed an independent relation between SAF and both micro- and macrovascular complications in T2DM –55. Ahdiet al.  demonstr [et al.  confirme [et al.  showed tet al. [Regarding diabetic retinopathy, two Japanese studies specifically investigated the relationship between SAF and the severity of the disease in T2DM , 57. BotIn T1DM, four  studies reported the relation of SAF with retinopathy, nephropathy, and/or neuropathy in the past 5 years , 58–60. In summary, these recent reports  show consistent evidence of an association between SAF and end-organ complications in diabetes. Several studies have now confirmed that SAF is associated with retinopathy, even after correction for age and nephropathy. In addition, a few studies established an independent association between SAF and vascular complications in diabetes patients of Asian descent. However, no relation was found between SAF and vascular complications in diabetes patients with darker skin colour types. Unfortunately, none of the recent studies had a prospective design, but from earlier studies, some evidence remains for the predictive value of SAF in the development of diabetes complications in T2DM , 50. Theet al. [et al. [et al. [et al. [Three  studies have been performed in which the association between SIF and diabetes complications is investigated –63. All et al.  reported [et al.  showed tet al. [Supporting the association between SAF and cardiovascular complications, SAF has been related to several clinical markers of cardiovascular disease: diastolic function, arterial compliance, markers of endothelial function, and markers of atherosclerosis. A relation between SAF and functional anatomical changes, reflected in diastolic dysfunction, was first shown in a study by Hartog et al.  in patieet al. , 65.et al. [et al. [et al. [et al. [et al. [et al. [Several studies have shown that SAF is related to arterial compliance evaluated by arterial pulse wave velocity (aPWV). Ueno et al.  and Janu [et al.  describe [et al. , de Groo [et al. , and Lla [et al. . In acco [et al.  recentlyet al. [et al. [Relations between SAF and markers of microvascular and endothelial function have been described mainly in diabetes and renal failure patients. Araszkiewicz et al.  found a  [et al.  observedIn earlier studies, it had already been shown that AGEs measured in skin samples using collagen fluorescence are associated with coronary calcium score, as a surrogate marker for coronary atherosclerosis. More recently, SAF has been reported to be associated with clinical markers of atherosclerosis, such as intima media thickness and coronary calcification score, by several groups , 73–75. Vascular complications are common and receive much attention in diabetes research. More recently, diabetes have also been linked to other disorders, such as impaired cognitive function, Alzheimer’s disease , and depet al. [et al. [Spauwen et al.  reportedet al. . Moran e [et al.  demonstret al. [Van Dooren et al.  showed tDiabetes has been shown to be a risk factor for retinal detachment after cataract surgery . In a piAGE measurements, SAF in particular, have been included in some large cohorts, such as the Maastricht study  (enricheIt has been extensively shown that skin fluorescence is associated with a wide variety of long-term complications in diabetes mellitus. This association is supported by the relation between skin fluorescence and AGE accumulation in several tissues with slow turnover, between skin fluorescence and past long-term glycaemic control, and between skin fluorescence and clinical markers of cardiovascular disease. A limited number of studies have also provided evidence for an independent predictive role of skin fluorescence for cardiovascular complications and mortality in diabetes. More prospective studies, with longer follow-up period and larger group size, are being conducted to establish the predictive role and potential benefit of skin fluorescence in disease management of diabetes patients.


In recent years, PCR has been become widely applied for the detection of trypanosomes overcoming many of the constraints of parasitological and serological techniques, being highly sensitive and specific for trypanosome detection. Individual species-specific multi-copy trypanosome DNA sequences can be targeted to identify parasites. Highly conserved ribosomal RNA (rRNA) genes are also useful for comparisons between closely related species. The internal transcribed spacer regions (ITS) in particular are relatively small, show variability among related species and are flanked by highly conserved segments to which PCR primers can be designed. Individual variations in inter-species length makes the ITS region a useful marker for identification of multiple trypanosome species within a sample.Trypanosoma congolense, Trypanosoma brucei and Trypanosoma vivax and compared to a modified (using eluate extracted using chelex) ITS-PCR reaction.Six hundred blood samples from cattle collected in Uganda on FTA cards were screened using individual species-specific primers for Trypanozoon, a prevalence of 10.5% was observed as compared to 0.2% using ITS PCR (Kappa = 0.03). For Trypanosoma congolense, the species-specific PCR reaction indicated a prevalence of 0% compared to 2.2% using ITS PCR (Kappa = 0). For T. vivax, species-specific PCR detected prevalence of 5.7% compared to 2.8% for ITS PCR (Kappa = 0.29).The comparative analysis showed that the species-specific primer sets showed poor agreement with the ITS primer set. Using species-specific PCR for Trypanozoon (T. b. brucei s.l). While ITS primers are useful for studying the prevalence of trypanosomes causing nagana (in this study the species-specific primers did not detect the presence of T. congolense) there were discrepancies between both the species-specific primers and ITS for the detection of T. vivax.When selecting PCR based tools to apply to epidemiological surveys for generation of prevalence data for animal trypanosomiasis, it is recommended that species-specific primers are used, being the most sensitive diagnostic tool for screening samples to identify members of  Routine diagnosis of trypanosomiasis using classical parasitological approaches shows poor sensitivity under field conditions . This isTrypanosoma brucei s.l., Trypanosoma congolense and Trypanosoma vivax) and PCR based methods for their amplification developed.In recent years, PCR has been widely applied for the detection of trypanosomes and has been shown to be highly sensitive and specific in the laboratory . The useTrypanozoon , the most common target is the 177 bp DNA satellite repeat sequence originally described by Sloof et al. [Trypanozoon genomic DNA, were able to detect 0.1 pg of parasite DNA, i.e. the amount of DNA equated to that present in a single trypanosome [et al. [Trypanozoon are used to identify T. b. brucei as a first stage in the process of identifying human infective parasites within an animal population. To further discriminate T. b. brucei from T. b. rhodesiense PCR reactions targeting the single copy serum-resistance-associated (SRA) gene [SRA gene as well as the gene for phospholipase C (GPI-PLC) was developed [T. b. rhodesiense, but only GPI-PLC is present in T. b. brucei, so if a sample shows positive amplification for GPI-PLC this indicates that sufficient T. brucei s.l. genomic material is present to detect a single copy gene while the presence or absence of SRA determines whether T. b. rhodesiense is present.For detection of f et al.  that exipanosome . Weaker  [et al.  found thRA) gene  have beeRA) gene . To permeveloped . Both ofTrypanosoma congolense (savannah) is a highly pathogenic trypanosome that is the most widespread, in terms of both geographical and host range across Sub Saharan Africa [T. congolense, which target a satellite sequence of 316 bp have been developed [T. congolense savannah was 0.1 pg of parasite DNA, again the amount of DNA in a single trypanosome and weak bands containing the amplification product could be detected when the DNA content was 0.01 pg or an estimated 1/10th that of a single parasite [n Africa . Specifieveloped ,14. The parasite .Trypanosoma vivax can be identified using universal primers targeting a fragment of the gene encoding T. vivax specific antigen [T. vivax isolates from diverse geographic locations in Africa and South America. antigen ,16; thisThese species-specific primers show good sensitivity and specificity against their target species. However, to screen large numbers of samples such as are derived from naturally infected hosts to determine prevalence data, multiple PCRs need to be undertaken for each sample which is both time consuming and expensive. It would be desirable to be able to apply a single PCR reaction that could simultaneously differentiate all of the trypanosome species present within a sample, making screening of a large number of samples viable in terms of time and cost.et al. [et al. [et al. [et al. [et al. [et al. [et al. [Trypanozoon and T. congolense but reported more T. vivax positives using the ITS primer sets than were derived using the species-specific primers. The species specific primer set had been developed for West African T. vivax isolates and this could explain the lower levels of positives generated and therefore the disagreement between the ITS PCR tests and species-specific PCR when applied to East African samples. In disagreement with the work of Thumbi et al. [et al. [et al. [Ribosomal RNA (rRNA) genes are highly conserved and have been proved useful for comparisons of closely related species. Eukaryotic rRNA genes occur as tandem repeats of units separated by a non-transcribed spacer (NTS) region and internal transcribed spacer regions (ITS); the ITS regions are relatively small, show variability among related species and are flanked by highly conserved segments to which PCR primers can be designed. Individual variations in inter-species length makes the ITS region a useful marker for species differentiation in trypanosomes and this and their high copy number, of around 200 makes the (ITS) a useful target for species differentiation in trypanosomes and other species ,18. Njiret al.  and Cox  [et al.  is a sin [et al.  compared [et al.  and Cox i et al.  a large i et al. , showed  [et al.  were siget al. [et al. [Although FTA cards present an efficient method of collection, storage and transportation of field material using one punch to seed the PCR reaction can reduce the chances of a positive PCR results . This iset al.  have rec [et al.  to incluTrypanozoon[T. congolense[T. vivax[et al. [In this study we have extensively tested a substantive number of cattle samples from Uganda to compare the application of species-specific primer sets targeting ypanozoon,12, T. congolense and T. v[T. vivax,15 with x[et al. .A total of 600 blood samples were available for testing that had been collected from cattle in Uganda in 2006 from baseline sampling for the Stamp out Sleeping Sickness campaign . SamplesAt the national and district levels, the study was conducted with the approval of the Coordinating Office for Control of Trypanosomiasis in Uganda (COCTU) as well as the District Veterinary Officers (DVOs) in each of the study districts.et al. [et al. [et al. [et al. [In order to allow comparison between the species-specific and the ITS-PCR of Cox et al.  the prot [et al. . Ten dis [et al. . For PCR [et al.  for both [et al. . The secTrypanozoon DNA , T. con TCK-PCR ,14) and  (TV-PCR ). The ba [et al.  are showT. brucei s.l was used. Negative controls consisting of water only instead of eluate and washed blank discs were run with each PCR.One positive control (genomic DNA) for each trypanosome species was used in the corresponding species specific PCR, while for ITS-PCR, a positive control of Amplification products were resolved in 1% (w/v) agarose gels along with 100 bp molecular weight Superladder . The agarose gel was prepared in 1 × TBE   stained with 5 μM ethidium bromide. The gels were run in 1xTBE, 5 μM ethidium bromide for at least 45 minutes at 100 volts and visualized under an ultra violet transilluminator .When the amplification reaction gave a signal of the expected size according to the set of primers used without any signal in the negative control, infection was considered to be confirmed.http://www.statstodo.com/CohenKappa_Pgm.php) was used to compare the three PCR reactions targeting the Trypanozoon. All other Kappa tests were conducted using WinPepi  with a ion 3.15 .Trypanozoon, species-specific PCR using TBR detected 63  = 8.2%-13.2%) parasitic events, GPI-PLC-PCR detected 46  parasitic events while ITS-PCR detected 1  parasitic events  parasitic events while ITS-PCR detected 13 . The species-specific reactions  and ITS-PCR did not agree positively on any samples with ITS-PCR identifying 13 positives in total. Kappa testing showed poor agreement between the species-specific reactions  and ITS-PCR .Comparing the sensitivity  parasitic events while ITS-PCR detected 17  parasitic events. TV-PCR and ITS-PCR only agreed positively on eight animals with TV-PCR and ITS-PCR identifying a further 26 and nine positives respectively. Kappa testing showed poor agreement between TV-PCR and ITS-PCR .Comparing the sensitivity  infected with T. theileri. A species-specific PCR reaction that targets T. theileri satellite sequence of 500 bp size was developed by Rodrigues et al. [T. theileri DNA (data not shown).In addition to these pathogenic trypanosome species, ITS-PCR was able to identify non-pathogenic s et al.  and was et al. [Trypanozoon it performed much better for the detection of T. congolense when compared to species-specific reactions.The use of ITS-PCR in diagnosis of African trypanosomes allows the identification of several trypanosome species in the same reaction. This is both a saving in time and cost, as less PCR reactions need to be carried out to gain an understanding of the prevalence of trypanosomes in an area. Although FTA cards have simplified the collection and transport of blood samples , often Det al.  modifiedTrypanozoon while the species-specific reaction diagnosed 63 parasitic events. The difference between the sensitivity of ITS-PCR and species-specific PCR reactions observed here might relate to the frequency of the target within the parasite genome. The copy number of ITS target in the genome is 100–200 compared to 10,000 for TBR and the higher sensitivity of the latter reaction could be due to the increased copy number of the target DNA. However, good agreement was seen between TBR and GPI-PLC; with the latter targeting a single copy gene [et al. [Trypanozoon DNA while TBR-PCR identified 21% (n = 245) of samples; these samples were collected on FTA®cards from cattle in Uganda but PCR was conducted directly on the discs after washing. However, more research by de Clare Bronsvoort et al. [et al. [et al. [et al. [et al. [et al. [Trypanozoon (1.9-3.9%).In this study, ITS-PCR identified only one (0.2%) sample to contain opy gene  it wouldt et al.  had show [et al.  showed a 21% n = 5 of samp [et al.  reported [et al.  extracte [et al. , the speT. congolense savannah type, T. congolense forest type and T. congolense kilifi type using ITS PCR even after prolonged separation time of the bands by electrophoresis. This was attributed to the band sizes of the amplified products of the three types are close to each other . For this reason, any product with a band size above 1400 bp was classified as T. congolense species. The PCR reactions specific for T. congolense savannah, T. congolense forest and T. congolense kilifi were negative for all the examined samples with Kappa testing suggesting poor agreement between the two diagnostic PCR tests. Previously Cox et al. [T. congolense while only one sample was diagnosed using specific reactions for T. congolense savannah. The inability of the species-specific PCR reactions to identify the T. congolense species diagnosed with ITS-PCR could be explained by the existence of a new T. congolense sub-species which is similar to T. congolense savannah and T. congolense forest in ITS target, while there is no specific reaction to identify these isolates at present. Isoenzyme analysis by Gashumba et al. [T. congolense from Uganda was separated from other T. congolense isolates suggesting that further analysis of T. congolense from Uganda may be required.It was difficult to differentiate between x et al.  had compa et al.  had prevT. vivax specific antigen were used for the diagnosis of T. vivax species. The universal primer set diagnosed 34 T. vivax infections while ITS-PCR identified 17 positive samples. Another set of primers targeting satellite DNA sequence were used and none of the samples were positive with these primers (data not shown). This was not unexpected since the primers targeting the satellite sequence are not present in some isolates of T. vivax[et al. [T. vivax infections in cattle blood samples using the primer set amplifying the satellite sequence, while ITS-PCR identified six T. vivax infections. Although Cox et al. [T. vivax may not accurately assess the level of infection within wild animal samples from Tanzania [In the present study, universal primers targeting the gene encoding  T. vivax. Cox et x[et al.  identifix et al.  found liT. theileri was identified in 7.6% of samples using the modified nested ITS PCR. The prevalence obtained was similar to that obtained by Cox et al. [Trypanozoon prevalence village (3%) but lower than that seen in a high Trypanozoon prevalence village (47%). As there were no positive results using the species-specific PCR reaction targeting T. theileri it was difficult to gauge the sensitivity of the modified ITS PCR in the current work.x et al.  in a lowTrypanozoon within cattle populations, on samples that are collected on to FTA cards (that includes the zoonotic T. b. rhodesiense) species-specific primers are more suitable in terms of sensitivity. However, as species-specific primers did not identify any T. congolense and there were differences in the amplification of T. vivax, the ITS reactions of Cox et al. [et al. [The present study suggests that for estimating prevalence of x et al.  and othe [et al. ) might bThe authors declare they have no competing interests and the sponsors had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.HAA carried out the study, the molecular approaches and participated in preparing the manuscript. KP and SCW designed the study supervised the molecular approaches and participated in preparing and editing the manuscript. ETM carried out the statistical analyses and participated in preparing and editing the manuscript. All authors read and approved the final version of the manuscript.


Using ultrasonic devices in endodontics can enhance the antibacterial and tissue dissolving ability of different root canal irrigants such as sodium hypochlorite (NaOCl) which is the most common irrigant with excellent antibacterial and tissue dissolving abilities. However, due to its high surface tension, its penetration into the irregularities of the root canal system is a challenge. The purpose of this paper was to review the different ultrasonic devices, different types of ultrasonic irrigation, the effect(s) of ultrasonic activation on the antibacterial and biofilm-removal abilities of NaOCl as well as the effect of ultrasonic activation on the smear layer removal ability of NaOCl. Accordiet al. , only 40et al. , 4. Enterococcus faecalis  can penetrate the dentinal tubules of root canal walls up to 800-1000 µm deep [°C [The aim of root canal irrigation is to remove the pulp tissue remnants and microorganisms (in either planktonic or biofilm forms) , eliminadeep [°C .Active and passive root canal irrigationPassive irrigation is conducted by slow dispensing of the irrigant of choice into a canal through a variety of different gauged needles [ needles . In orde needles . Passive needles .active irrigation initiates dynamics and flow within the fluid and thus improves root canal disinfection. In well-shaped canals, fluid activation has a critical role in cleaning and disinfection of the canal irregularities by facilitating the fluid penetration through all aspects of the root canal system [On the other hand, l system , 11. Physics of ultrasonicUltrasound is a vibration or acoustic wave with similar nature as sound but with a frequency higher than the highest frequency detectable by the human ear (approximately 20000 Hz) . Ultrasomagnetostriction that converts the electromagnetic energy into mechanical energy. The second method works according to the piezoelectric principle and uses a crystal which changes in size by applying electrical charge [There are two basic methods for producing an ultrasonic wave. First is l charge , 15. TheMagnetostrictive units have two major drawbacks for endodontic application. First they have elliptical movement and oscillate in a figure-eight manner and second, they generate heat, so adequate cooling is required. vs. 24 in magnetostrictive devices). The other advantage is the piston-like linear movement of tip in piezoelectric units from back to front which is ideal for endodontic treatment [One major advantage of piezoelectric units over magnetostrictive devices are production of more cycles per second ”, “Ultrasonics Activation AND Sodium Hypochlorite (47 articles)", "Ultrasonics AND NaOCl (51 articles)”, “Passive Ultrasonic Activation AND Sodium Hypochlorite (24 articles)”.Effects of ultrasonic irrigation in endodonticsUsing ultrasonic energy in endodontic treatment has improved treatment quality in many aspects, including access to root canal entry holes, cleaning, shaping and filling the canals, eliminating the obstructions and intracanal materials and endodontic surgery . Ultrasonic devices can be utilized in two manners; simultaneous combination of ultrasonic irrigation/instrumentation and passive ultrasonic irrigation (PUI) , 18. BecApplying ultrasound for passive irrigation seems more advantageous , 21. ForEffect of ultrasonic energy on antibacterial activity of NaOClNaOCl is the most common root canal irrigant with excellent antibacterial and tissue dissolving abilities . Irrigatin vitro study by Tardivo et al. [E. faecalis. Huque et al. [et al. [et al. [In an o et al.  there wae et al.  showed t [et al.  and Siqu [et al.  have indUltrasonics and bacterial biofilmset al. [E. faecalis biofilms. Harrison et al. [According to Bhuva et al.  both conn et al.  concludeet al. [E. faecalis biofilm. Neelakantan et al. [E. faecalis biofilm compared to the ultrasonic. Bhardwaj et al.  showed tn et al.  showed tEffect of ultrasonic on smear layer removalet al. [Ahmad et al.  claimed et al.  showed tet al.  also comet al. -37.et al. [et al. [Researchers who found the cleaning effects of ultrasonic beneficial, used the technique only for the final irrigation of root canal after completion of hand instrumentation , 38, 39.et al. , 40 clai [et al.  recommenet al. [Prati et al.  also menet al. , 44 showet al. [Baumgartner and Cuenin  also obset al.  evaluateet al. [et al. [et al. [in vitro model. Findings showed that both activation techniques are important adjuncts in removing the smear layer.Mozo et al.  showed t [et al.  showed t [et al.  comparedCurtis and Sedgley  showed tet al. [et al. [2O2. Superiority of ultrasonication of the intra-canal irrigant over the manual technique in removing the smear layer was demonstrated by Ribeiro et al. [Kocani et al.  showed t [et al.  showed to et al. . et al. [et al. [Blank-Goncalves et al.  showed t [et al.  ultrasonet al. [et al. [Paque et al.  confirme [et al.  assessedUltrasonics vs. sonic irrigation Sonic instruments use a lower frequency (1000-6000 Hz) compared to ultrasonic instruments (25000 Hz). In both types of instruments the file is connected at an angle of 60-90 degrees to the long axis of the handpiece. However, the vibration pattern of ultrasonic files is different from that of sonic instruments. Ultrasonically activated files have numerous nodes and antinodes across the length of the instrument, whereas sonic files have a single node near the attachment of the file and one antinode at the tip of the instrument. Sonic instruments produce an elliptic, lateral movement, similar to that of ultrasonic files , 17.°C (in areas close to the tip of the instrument) and 37°C (away from the tip) when the irrigant was ultrasonically activated for 30 sec without replenishment. A cooling effect from 37°C to 29°C was recorded when the irrigant was replenished with a continuous flow of irrigant. The temperature of the irrigant was 25°C. The external temperature stabilized at 32°C during a continuous flow of the irrigant and reached a maximum of 40°C in 30 sec without continuous flow. Ahmad [°C-rise of temperature during a continuous flow of irrigant. The initial temperature of the irrigant was 20°C. A rise of temperature within these ranges will not cause pathological temperature rises in the periodontal ligament.Cameron  reportedw. Ahmad  reported1. Superiority of ultrasonic irrigation with NaOCl over passive irrigation with syringe is still controversial.2. Superiority of ultrasonic activation of NaOCl on endodontic biofilms over other irrigation methods is controversial.3. Superiority of ultrasonic activation of NaOCl on smear layer removal is controversial.


Iron is essential for the Central Nervous System development, i.e. mielinization process and cellular differentiation as well as correct functioning of neurotransmitters . Preterm


Aptamers are a class of therapeutic oligonucleotides that form specific three-dimensional structures that are dictated by their sequences. They are typically generated by an iterative screening process of complex nucleic acid libraries employing a process termed Systemic Evolution of Ligands by Exponential Enrichment (SELEX). SELEX has traditionally been performed using purified proteins, and cell surface receptors may be challenging to purify in their properly folded and modified conformations. Therefore, relatively few aptamers have been generated that bind cell surface receptors. However, improvements in recombinant fusion protein technology have increased the availability of receptor extracellular domains as purified protein targets, and the development of cell-based selection techniques has allowed selection against surface proteins in their native configuration on the cell surface. With cell-based selection, a specific protein target is not always chosen, but selection is performed against a target cell type with the goal of letting the aptamer choose the target. Several studies have demonstrated that aptamers that bind cell surface receptors may have functions other than just blocking receptor-ligand interactions. All cell surface proteins cycle intracellularly to some extent, and many surface receptors are actively internalized in response to ligand binding. Therefore, aptamers that bind cell surface receptors have been exploited for the delivery of a variety of cargoes into cells. This review focuses on recent progress and current challenges in the field of aptamer-mediated delivery. In contrast to antisense oligonucleotides and small interfering RNAs (siRNAs) that inhibit translation of proteins by Watson-Crick base-pairing to their respective messenger RNAs, aptamers bind to existing proteins  with high affinity and specificity, analogous to monoclonal antibodies. Aptamers are typically generated by an iterative screening process of complex nucleic acid libraries (>10 (SELEX) . The SELO-methyl pyrimidines [RNA and DNA aptamers both have theoretical advantages and proponents, but aptamers of comparable affinity and specificity can be generated from RNA or DNA. Since nuclease resistance is critical for aptamer stability in biological fluids, RNA libraries employed in SELEX are front-loaded with 2'-modified nucleotides, most commonly 2'-fluoro- or 2'-imidines . Probablimidines , but modimidines .in vitro transcription for testing in vitro and in certain small scale animal models. However, chemical synthesis is necessary for larger scale applications. Since the efficiency of chemical synthesis decreases with oligonucleotide length, it is often necessary to “minimize” or “truncate” an aptamer prior to study in vivo. Although the technology continues to improve, aptamers longer than 60 nts are not currently amenable to cost-effective chemical synthesis, and the shorter, the better. Due to the relatively small size (8 kDa to 15 kDa) of truncated aptamers, their circulating half-lives are limited not by plasma stability but by renal clearance, which can be improved by conjugation to high molecular weight groups such as polyethylene glycol (PEG) [Most SELEX libraries have random regions ranging from 20 to 60 nucleotides (nts), flanked by constant regions for amplification and transcription, and therefore total lengths ranging from 70 to greater than 100 nts. Sufficient quantities of any length aptamer can be generated by Over the past two decades, this technology has enabled the generation of aptamers to a myriad of proteins including reverse transcriptases, proteases, cell adhesion molecules, infectious viral particles, and growth factors .SELEX has traditionally been performed using purified proteins, and cell surface receptors may be challenging to purify in their properly folded and modified conformations. Therefore, relatively few aptamers have been generated that bind cell surface receptors. However, improvements in recombinant fusion protein technology have increased the availability of receptor extracellular domains as purified protein targets, and the development of cell-based selection techniques has allowed selection against surface proteins in their native configuration on the cell surface. With cell-based selection, a specific protein target is not always chosen, but selection may also be performed against a target cell type with the goal of letting the aptamer choose the target. The past decade has seen the generation of several aptamers that bind to cell surface receptors ,20,21,22et al. [99mTc. In order to assess the tumor uptake and biodistribution property of the radiolabeled anti-TN-C aptamer it was injected intravenously into mice bearing gliobastoma (U251) and breast cancer (MDA-MB-435) tumor xenografts. Scintigraphic images of the tumors were taken 18 hours after injection and revealed that the aptamer was exclusively localized in tumors. This was achieved due to the combined effect of the efficient uptake of aptamer by the tumor and its rapid clearance from the blood [Tenascin-C (TN-C) is an extracellular matrix protein (ECM) that is implicated in the process of tissue remodeling. It is overexpressed in the tumor stroma where it is thought to enhance angiogenesis and invasion ,41. In oet al.  performeet al. . A truncbastoma U1 and breet al. [N-acetylated-α-linked-acidic-dipeptidase) of PSMA. A truncated version of A10 was created that bound to a PSMA-positive (LNCaP) prostate cancer cell line. The binding was selective, because A10 did not bind the PSMA-negative PC3 prostate cancer cell line. Prostate-specific membrane antigen (PSMA) is a type II membrane-associated metallopeptidase that is overexpressed on the surface of prostate cancer cells. PSMA is also expressed in the vasculature of many other solid tumors . Therefoet al.  selectedBased on these findings and the previous knowledge that PSMA is internalized via clathrin-coated pits to endosome , three det al. [in vitro application. This was the first demonstration that aptamers could be used to deliver lethal siRNAs to targeted cancerous cells. These groups utilized different strategies to accomplish this goal. Chu et al.  used a bet al. [in vivo efficacy of this delivery approach, the authors directly injected the chimera into LNCaP tumors that were established in mice. As a control, the same application was performed on mice bearing PC3 (PSMA-negative) tumors. Mice bearing the LNCaP tumors showed reduction in the tumor volume due to the chimera injection (but not the injection control chimeras), whereas no effect was observed in the PC3 tumors. This study clearly demonstrated the specificity and efficacy of the aptamer mediated siRNA delivery approach. In a recent study done by the same group [McNamara et al.  also demme group , the chiet al. [In a subsequent study by Wullner et al. , the autet al. [In situ immunofluorescence microscopic analysis and cytotoxic assay demonstrated that the conjugate was internalized into the cells and resulted in targeted cell killing. In parallel, a series of other studies demonstrated that PSMA aptamers could be exploited in a variety of ways to deliver cargo into cells. Gelonin is a ribosomal toxin that can inhibit the process of protein synthesis and is cytotoxic. However, it is membrane impermeable and needs an usher for its cellular entry. In one of the first examples of aptamer-mediated delivery, Chu et al.  realizedco-glycolic acid) PLGA were used to solve this targeting problem [In vitro cytotoxicity assays done with LNCaP cells demonstrated that the A10 conjugated Pt(IV) prodrug-PLGA-PEG-nanoparticle was more potent than either free cisplatin or nontargeted (aptamer non-conjugated) nanoparticles. Farokhzad et al. [Since direct conjugation limits the amount of cargo that each aptamer can deliver, other strategies have incorporated aptamers as targeting moieties for a variety of functional polymers and nanoparticles, each of which can carry many molecules. For example, tumor resistance to cytotoxic chemotherapeutic agents is due in part to insufficient delivery to and uptake by cancer cells. Biodegradable nanoparticles (NP) derived from poly, the causative agent of AIDS (Acquired Immuno Deficiency Syndrome) is an enveloped retrovirus ,49 that et al.  selectederaction  and neuteraction . Zhou et [et al.  of the R [et al. . To one et al. [Nucleolin, first described by Orrick et al.  is a preet al. . In addiet al. . Consequet al. . Moreoveet al. . Therefoet al. to have anti-proliferative activity and subsequently found to bind nucleolin. This aptamer was therefore not “selected” in the way that most of the other aptamers described in this review were. It was further established that AS1411 inhibited the pro-survival NF-κB signaling pathway [et al. Anti-nucleolin aptamer was found to inhibit the binding of nucleolin to Bcl-2 mRNA. This resulted in the destabilization of the mRNA with a consequent decrease in the level of anti-apoptotic Bcl-2 protein in the breast cancer cells [et al. [in vivo. Currently, the anti-nucleolin aptamer AS1411 is in phase II clinical trails for acute myeloid leukemia and renal cell carcinoma. A 26-nucleotide guanosine-rich (G-rich) DNA sequence (AS1411) was discovered serendipitously by Bates  pathway  and thuser cells . Studies [et al.  in nude et al. [et al. used radio-labeled F3-peptide to deliver α-particle emitting 213Bi isotope into the nucleus of tumor cells in the mouse intraperitonial xenografts model [In addition to the direct anti-cancer effects of AS1411, the fact that nucleolin shuttles between the cell surface, cytoplasm, and nucleus in rapidly dividing cells means that, in principle, nucleolin can be used to deliver cargo into cancer cells. A precedent for this has already been set. A 34-amino acid peptide (F3) discovered by Christian et al.  using a ts model . Reductiet al. who conjugated it with a multimodal nanoparticle (MFR-AS1411) and monitored uptake into C6-rat glioma cells by using fluorescence confocal microscopy. For the in vivo tracking of MFR-AS1411, it was injected systemically into nude mice bearing C6 tumor xenografts subjected to both whole body scintigraphic and Magnetic Resonance (MR) imaging techniques. Using these in vivo and in vitro methods, these authors have demonstrated that the anti-nucleolin aptamer can target nanoparticles to cancer cells expressing nucleolin on their cell surface and can potentially be used as a non-invasive imaging tool for the diagnosis of cancer [et al. [In vitro studies demonstrated that the aptamer-TMPyP4 complex was readily taken up by MCF7 cells expressing the nucleolin and were severely damaged in response to photodynamic therapy (PDT) as compared to the control normal epithelial cells. Given the observation that AS1411 is internalized at much lower concentrations than those necessary for anti-proliferative effects [However, the unusually high serum stability and low immunogenicity of the anti-nucleloin aptamer AS1411  make it f cancer . Recentl [et al.  used AS1 effects , AS1411 +3 binds TfR, the transferrin-Fe+3-TfR complex undergoes endocytosis and is transported to the endosomal compartment where the iron is released and the TfR-apotransferrin is recycled back to the plasma membrane [et al. [TfR is a ubiquitously-expressed membrane bound protein that is involved in the process of iron uptake into cells and thus maintains cellular iron homeostasis. After transferrin-Femembrane ,66. To t [et al.  selected [et al. . PrompteO-glycosylation. Aberrant and incomplete glycosylation of MUC-1 is often associated with various epithelial cancer cells . These abnormally glycosylated proteins, termed as glycoforms, represent a valuable class of tumor biomarkers because they are expressed only on cancer cells and are distinct from those expressed on normal cells [et al. [e6 was coupled to the 5’ amino group of the MUC-1 aptamers and targeted to cancer cells expressing the aberrant MUC-1 glycoforms. These conjugated MUC-1 aptamers produced cytotoxic singlet oxygen species upon photodynamic therapy (PDT) and displayed greater than 500 times enhanced cellular toxicity as compared to when chlorin e6 was used as a free drug. Interestingly, normal human mammary cells that express the fully glycosylated MUC-1 were not affected by the conjugated MUC-1 aptamers upon PDT thus demonstrating remarkable cancer cell-targeting specificity of these aptamers [et al. [2) and subsequently radio-labeled them with the isotope 99mTc. The 99mTc-MAG2-conjugated aptamers were injected systemically into mice bearing MCF7 (breast cancer cell) xenografts, and the bio-distribution was studied. The aptamer-radionuclide was taken up by tumor cells suggesting these aptamers may also be useful for the diagnosis and staging of breast cancer. MUC-1 is a cell surface associated glycoprotein that is extensively modified by al cells . Studiesal cells . Aptamer [et al.  selectedaptamers . Pieve e [et al.  from theet al. [et al. [et al. [PTK7 is a membrane bound receptor tyrosine kinase-like molecule. It is over-expressed in colon carcinomas and is also known as colon carcinoma kinase-4. Although it contains a catalytically inactive tyrosine kinase domain, it has been suggested to retain a role as a signal transducer in some tumors types . A DNA aet al. . Howeveret al. . Using pet al. . In subset al. ,73,74. I [et al.  recentlyet al. [50 of sgc8c-Dox was found to be same as free-Dox. The sgc8c aptamer is therefore a promising candidate for targeted drug delivery. Huang et al.  covalentet al. [et al. [More recently, Taghdisi et al.  used the [et al.  recentlyAnthracycline family-based chemotherapeutic drugs are membrane permeable and are randomly taken up by the cells through the process of passive diffusion. However, conjugating them with the sgc8c aptamer restricts their entry into cells that express PTK7. This “sieve” mechanism should curb the non-specific uptake of chemotherapeutic drugs and minimize the toxic effects of chemotherapeutic agents on normal cells.et al. [Immunoglobin heavy mu chain (IGHM) is the large polypeptide subunit of the IgM antibody. IGHM expression level on premature B-cell correlates with the development of Burkitt’s lymphoma ,78. As det al.  with theet al. . This pret al. [in vivo stability issues; therefore, these authors have also generated a modified RNA aptamer against EGFR that is currently being tested .EGFR is a transmembrane receptor tyrosine kinase and is considered the “prototype” for receptor-mediated endocytosis. Binding of EGFR to its cognate ligand causes receptor dimerization leading to autophosphorylation, internalization of the receptor, and activation of intra-cellular signal transduction pathways . EGFR ovet al.  recentlyAptamers are nucleic acid ligands with several properties that make them attractive as pharmaceutical agents. Aptamers bind their targets with high affinity and specificity and are amenable to large-scale chemical synthesis. The versatility of the aptamer selection process has facilitated the generation of aptamers that bind a wide array of targets, including several cell surface receptors. Aptamers which bind cell surface receptors that are internalized have been exploited to deliver a variety of cargoes into cells. Aptamers therefore may be used to deliver molecules that are not otherwise taken up efficiently by cells  or to limit delivery of molecules that are efficiently taken up by cells  to cells that express aptamer targets.in vivo SELEX techniques should facilitate the identification of additional cell type-specific targets and aptamers. The perfect target for aptamer-mediated delivery is one that is highly expressed on all target cells, is efficiently internalized, and is not expressed on the surface of non-target cells. Although a perfect target may not exist, several aptamers against excellent targets have been identified. Studies using these aptamers have provided “proof of concept” that aptamers can mediate cell type-specific delivery. Cargoes have included enzymes, toxins, chemotherapeutic agents, imaging agents, and siRNAs. Current challenges are both to optimize the cargo (what to attach and how to attach it) and to identify even better targets (and aptamers that bind them). With respect to the former, the fields of nanotechnology and RNA interference are rapidly maturing and will result in even more sophisticated aptamer constructs. With respect to the latter, further refinement of cell-based SELEX and in vitro data, we have only a modest amount of in vivo animal data and—to date—no human data demonstrating that aptamer-mediated delivery is feasible. One factor that has potentially slowed the preclinical development of some aptamer therapeutics is the cost of synthesis, which may be prohibitive for aptamers longer than 40–50 nucleotides, particularly if the application requires repeated and/or systemic delivery. However, as methods for the chemical synthesis of olignonucleotides have improved, the yields and costs of synthesis have improved. The synthesis of oligonucleotides is also very “scalable”, as evidenced by the growing number of olignonucleotide therapeutics entering clinical trials. Furthermore, by using aptamers to deliver highly potent cargo, the amount of aptamer required (and therefore synthesis costs) may be significantly less than required for using aptamers as direct inhibitors. Therefore, we anticipate that aptamer-mediated delivery will prove to be feasible in vivo and that translation of this approach to human patients is a realistic goal for the near future. Meanwhile, although we have an abundance of promising


In the literature, there have been conflicting reports on an association between male baldness  and the incidence of coronary artery disease (CAD).In 1979, Cooke  identifiIn 2001, Rebora et al.  performeThese predominantly inconclusive studies from the previous century are counterbalanced by several other positive studies performed in the current century. In 2000, Lotufo et al.  showed, Interestingly, in 2005 Mansouri et al.  found anp < 0.05). This study implies that early-onset androgenetic alopecia in males is independently associated with CAD, though the mechanisms need to be investigated.Recently, two studies from the same group showed that early onset androgenetic alopecia in males was independently associated with CAD , 9. In 2In the current issue of the Netherlands Heart Journal, Sari et al.  investigThe present study by Sari et al.  providesAltogether, it goes too far to say that bald is beautiful, but for many men it might be reassuring that male baldness is not significantly associated with CAD.