Seroepidemiological surveys show that the prevalence of human herpesvirus 8 (HHV-8) infection mostly varies in various geographical areas and reflects the local incidence of classic and endemic KS, being widespread in sub-Saharan Africa and Mediterranean countries and uncommon in the USA and Northern Europe. In the Middle East only few populations, such as Ashkenazi and Sephardic groups in Israel, have been adequately evaluated for HHV-8 seroprevalence. Among Iranian population a striking higher seroprevalence of HHV8 has been reported among haemodialysis (16.9%), renal transplant recipients (25%) and HIV (45.7%) patients compared to blood donors (2%). Kaposi's sarcoma (KS) is the rarest cancer in Iran, with an annual age-standardized incidence varying from 0.10 to 0.17 per 100,000 in males and from 0.06 to 0.08 per 100,000 in females. KS, however, is one of the most important malignancies in Iranian renal transplanted patients affecting up to 2.4% of organ recipients. The epidemiology of HHV8 and KS in Iran needs further evaluation. While the high prevalence of HHV-8 antibodies in HIV positive and haemodialysis individuals may be attributed to high-risk sexual behavior and polytransfusions, respectively, unknown determinants may be responsible for high seroprevalence of HHV8 and high incidence of KS in solid organ recipients. A global survey on HHV8 seroprevalence in Iran is mandatory to define co-factors associated with HHV8 infection and KS risk in the general Iranian population and in specific patient groups. Kaposi's sarcoma (KS) is a mesenchymal tumor involving blood and lymphatic vessels that was first described in Eastern Europe in the late 19th century : 1) ClasIn 1994 Chang et al. identified fragments of the Kaposi's sarcoma-associated herpesvirus (KSHV) genome in epidemic KS tissues, subsequently called human herpesvirus type 8 (HHV-8) . HHV-8 iSeroepidemiological surveys have shown that HHV-8 infection is not ubiquitous . The virFew studies have been performed on the HHV-8 distribution and incidence of Kaposi's sarcoma in Iran. This study aimed to recapitulate available data on the seroepidemiology of HHV-8 and incidence of KS in the Iranian population. A systematic review of the published articles from January 1980 to December 2010 was conducted to assess the seroprevalence of HHV-8 and the incidence of KS in Iran. Data were identified by searches of Medline, Current Contents, PubMed, and references from relevant articles, with the search terms "Kaposi's sarcoma" or "HHV-8", and "Iran".Data from the National Cancer Registry of Iran have reported that KS is the rarest cancer among Iranians . Only 50Worldwide KS accounts for only 0.02% to 0.07% of all malignancies in the general population . The regThe incidence of KS in kidney recipients has increased following an enormous increase in the number of kidney transplantations during the recent decades, particularly of those of Mediterranean descent . Post-trThe rate of chronic kidney disease and renal transplantations has increased during the last two decades in Iran -36. AbouKS is one of the most common cancers after renal transplantation in Iran  represenIn Iran, incidence of KS among renal transplants varied from 0.45% to 2.4% in different studies Table  37,38,4-43,52-55The KS incidence among Iranian transplants recipients had a peak during the first 2 years post transplantation. The time interval between transplantation and onset of KS was relatively early compared to other skin tumors. This observation is in agreement with other studies from the Middle East region reporting appearance of KS lesions in 6.5 to 27 months following kidney transplantation .The mean-age of Iranian renal transplants developing KS is below 50 years, which is lower than that of patients with classic Kaposi's sarcoma (50-79 years). This finding is consistent with data reported by other studies on renal transplant patients from different regions of the world ,45,60,61About 90% percent of transplant recipients affected by KS present cutaneous or mucosal lesions or both types. Visceral involvement occurred in 25% to 30% of kidney transplant patients . VisceraMost cases of post-transplantation KS develop as a result of viral reactivation, since more than 80 percent of transplant recipients with KS are seropositive for HHV-8 before transplantation ,64. Renath, 2004) at the Research Center of Iranian Blood Transfusion Organization. The 256 blood donors, enrolled without any obligations, represented the 25.6% of blood samples screened on that day. All sera were tested for the presence of antibodies against HHV-8 lytic antigens by HHV-8 IgG EIA and by HHV-8 IgG IFA commercial kits  as for manufacturer's instructions and each sample defined as "positive" if tested positive to both assays. The sensitivity of EIA and IFA tests was 90.4% and 100%, respectively, as reported in the manufacturer's catalogue. The specificity of EIA and IFA was 93% and 94%, respectively.Few serological survey on HHV-8 infection have been performed in Iran ,66. One P = 0.36) [Among the 256 healthy Iranian blood donors  only 5 , including 4 males and one female, were positive for HHV-8 antibodies. Conversely, among the 118 haemodialysis patients , 20  tested positive, including 8 males and 12 females. In the HIV positive group  16 out of the 35 patients , all males, were positive for HHV-8 . Also, f = 0.36) . Crude oLow prevalence of HHV-8 in blood donors may indicate that virus is not widespread in this population; however, this study is not sufficient to determine the extrapolation of the true prevalence of HHV-8 in Iranian healthy population. Blood donors in above mentioned study were only from Tehran and it may be representative of the prevalence of HHV-8 in Tehran rather than Iran. The distribution of this virus probably is not ubiquitous in all over the Iran, so it is recommended to perform surveillance studies on different populations living in different areas of Iran in order to determine the global seroprevalence of HHV-8. Also blood donors commonly have not considered as persons with high risk behaviors. It is better that future study populations be general population rather than blood donors to estimate more relevant prevalence of HHV-8 infection.P = 0.02). In particular, 8 out of 17 (47%) patients were seropositive in the group older than 55 years, versus 17 out of 83 (20%) patients in the group younger than 55 years. There were no significant differences in HHV-8 seropositivity regarding sex. Seropositive and seronegative patients were followed for 16 months and only one HHV-8 seropositive patient (1/25) developed Kaposi's sarcoma [In another study by Ahmadpoor et al., 100 serum from Iranian renal transplant recipients  were analyzed for antibodies against the latent nuclear antigen of HHV-8  and 25%  sarcoma . The ris sarcoma ,67.Different rates of HHV-8 infection have been reported in various populations in the world. The prevalence of HHV8 in healthy individuals was found to be 1.3%-4.4% in Southeast Asia and the Caribbean regions and > 40% in Africa -70. In IIn conclusion, in Iran a high prevalence of HHV8 infection has been observed in several risk groups such as haemodialysis, renal transplant and HIV-positive patients. KS incidence among renal transplant patients is as high as that observed in transplanted patients from endemic regions for KS. A future study including a large population from different regions of Iran is needed in order to define co-factors associated with HHV8 infection and KS risk specific patient groups in Iran.The authors declare that they have no competing interests.SJ conceived the study and performed a critical review of original data on HHV8 and KS prevalence in Iran in 1980-2010 period. ZS, TM-A and RN contributed to KS prevalence analysis in Iran. AG participated in the study design and coordination. All authors read and approved the final manuscript.


P < 0.01). Small size (<10 mm) and patches found on endoscopy were significantly associated with FNRs (P < 0.05). Findings of submucosal tumor (SMT) with ulceration were significantly associated with true-positive results (P < 0.05). In conclusion, FNRs were found in 35.3% of GI-KS lesions and were especially related to the site of the esophagus and endoscopic early stage . An SMT with ulceration may be relatively easy to diagnose on endoscopic biopsy. Caution should be exercised when performing endoscopic biopsy of these lesions in AIDS patients and evaluating the histological features.Kaposi's sarcoma (KS) is a rare endothelial neoplasm mainly involving the skin, but it is often associated with AIDS. Diagnosis of gastrointestinal (GI) tract KS, a common site of visceral involvement in AIDS, is important, but endoscopic biopsy carries a risk of false-negative results (FNRs) due to its submucosal appearance. This study sought to determine the rate and causes of FNR for endoscopic biopsy of GI-KS lesions. Endoscopic biopsy samples of 116 GI-KS lesions were reviewed retrospectively. All GI-KS lesions were confirmed to be resolved following KS therapy. FNRs were yielded for 41 of the lesions (35.3%). Among upper and lower GI sites, the esophagus was the only site significantly associated with FNRs ( Kaposi's sarcoma (KS) is a cancer of the lymphatic and blood vessels that mainly involves the skin –3. It isKS can also involve the oral cavity, lymph nodes, and viscera –3, 8. ThThe purpose of this study was to determine the rate and causes of FNR from endoscopic biopsies of GI-KS lesions. Histopathology slides of endoscopic biopsy samples of 116 consecutive, GI-KS lesions from 24 HIV-infected patients who had not received anti-KS therapy were retrospectively reviewed. All biopsies were performed between 2002 and 2006 at the National Center for Global Health and Medicine (NCGM), a 900-bed hospital located in the Tokyo metropolitan area with the largest referral center for HIV/AIDS in Japan. The institutional review board at NCGM approved this study. All patients from whom clinical samples were obtained during endoscopic biopsy provided written informed consent prior to the procedure. Data obtained from the patient medical records was anonymized before analysis.HIV infection route was determined by the medical staff who interviewed each patient on the first visit to our hospital. Routes of HIV infection were determined by medical staff who questioned each patient face to face on the first visit to our hospital. Routes were classified into six categories: homosexual, bisexual, heterosexual, drug user, untreated blood products, and unknown. Patients who were homosexual or bisexual were regarded as men who have sex with men (MSM).+ cell counts were checked within 1 week of endoscopy. HIV-RNA viral loads (VLs) determined by real-time quantitative polymerase chain reaction (PCR) were reviewed within 1 month of endoscopy. The minimum detection level was 40 copies/mL of plasma. A positive result for real-time HIV-RNA was defined as ≥40 copies/mL. CD4Biopsy was performed using biopsy forceps . All biopsies were performed by well-trained endoscopists (experience of >1.000 colonoscopies). Negative results confirmed from biopsy samples for other GI diseases such as infection, inflammatory bowel disease, hyperplastic polyp, fundic gland polyp, inflammatory polyp, adenomatous polyp, angioectasia, GI lymphoma, premalignant lesion, esophageal cancer, gastric cancer, and colorectal cancer.Presence of proliferating spindle cells  with vasPositive response to KS therapy ; in particular, for visible GI-KS lesions without typical pathological findings from biopsy specimens, partial or complete resolution confirmed on follow-up endoscopy following KS therapy.A definitive diagnosis of GI-KS was defined as follows.GI-KS was evaluated in terms of site, size (≤10 mm or >10 mm), and macroscopic findings on endoscopy. Site of GI involvement was classified into 7 regions: esophagus, stomach, duodenum, ileum, right-side colon , left-side colon (descending colon and sigmoid colon), and rectum. Macroscopic findings were evaluated in the presence of reddish mucosa with patches , polypoiχ2 test. Those factors that emerged as significant (P < 0.10) on univariate analysis were included in a multiple exact logistic regression model. A final model was then developed by backward selection of factors showing values of P < 0.10 and odds ratios (ORs) and 95% confidence intervals (CIs) estimated.The descriptive patient characteristics were summarized, and the absence rate of spindle cells or vascular formations on pathology for the 116 samples was then analyzed to elucidate the FNR rate of endoscopic biopsy. To determine the cause of FNRs, the relationships between FNR and endoscopic findings  were evaluated using the P < 0.10 were considered significant. All statistical analyses were performed using Stata version 10 software .Values of + count was 71 cells/mL and median HIV-RNA VL was 115,000 copies/mL. GI symptoms were noted in 8 patients (33.3%) as follows: epigastric pain (n = 4), nausea or vomiting (n = 3), hematemesis (n = 1), melena (n = 1), hematochezia (n = 2), and diarrhea (n = 2) (duplicate data). Characteristics of the 24 patients are shown in n = 8, 20.0%) and right-side colon . Only one lesion with “Polypoid” appearance was noted in the terminal ileum. “SMT” appearance was noted in 46 lesions (39.7%) located mainly in the stomach  and duodenum . “SMT with ulcer” appearance was noted in 26 lesions (22.4%) located mainly in the stomach  and duodenum . “Bulky tumor” appearance was noted in 3 lesions only in the rectum.A total of 113 GI-KS lesions were from 24 HIV-infected patients . “PatcheNo clinical complications of GI-KS lesions were seen. There were no significant gastrointestinal bleeds or perforations, either spontaneous or after endoscopic biopsy. Diagnostic yield of GI-KS is shown in P < 0.01) associated with FNR. In regards to the size of lesions, those <10 mm in diameter were significantly associated with FNR (P < 0.05). As for macroscopic appearance, patches were significantly associated with FNR (P < 0.01), while a finding of SMT with ulceration was significantly associated with true positive results (P < 0.05).Among the GI locations, the esophagus was significantly  in this study. Previous studies on GI-KS patients have also reported a relatively low diagnostic yield for endoscopic biopsy , 23–25. Various clinical factors can contribute to FNR on biopsy. Submucosal location or tumor growth is considered to account for the poor diagnostic yield of standard forceps biopsies , 23. Morn = 4). Further prospective studies with a larger number of patients are needed.Secondly, with regard to the GI site, we found the esophagus to be the only site associated with FNR on both univariate and multivariate analyses. In this study, we assessed the site by dividing the GI into 7 parts. Because it is difficult to differentiate the ascending colon from the cecum and the descending colon from the sigmoid colon on endoscopy, we divided the colon into the right-side and left-side colon. No other study has divided the GI tract into small segments and investigated the diagnostic yield in such detail as in the present study. The yield of endoscopic biopsy in the upper GI tract has been estimated as 13%, which is low compared with 36% on sigmoidoscopy . It is nThirdly, in terms of the endoscopic appearance of GI-KS, small lesions (<10 mm) and patches were found to be significantly associated with FNR on univariate analysis. Because such characteristics could be confounding factors, we performed multivariate analysis and found that patches were correlated with FNR as an independent factor. We suggested that these lesions contain only a small amount of tumor tissue, which made such biopsy specimens too small to be diagnostically useful. On the other hand, the finding of “SMT with ulceration” was significantly associated with true-positive results, and these lesions were easily diagnosed by biopsy. This result may be attributable to the ulcerous appearance of the tumor, which makes it easy to obtain samples from the submucosal layer , 27.Fourth, we found that endoscopic biopsy is a safe diagnostic method for GI-KS even in the presence of an ulcerative or bulky tumor. A previous report  also higEndoscopists and clinicians should become familiar with characteristic endoscopic images of GI-KS and recognize that the diagnostic yield of GI-KS varies depending on the morphological features. In addition, pathologists should carefully evaluate lesions associated with FNR. Because KS-related GI lesions indicate visceral involvement, the indications for and selection of HAART and systemic chemotherapy need to be considered –10. GI-KEndoscopic biopsy is essential for diagnosing GI-KS and it is a safe method. While FNR were found in 35.3% of lesions, FNRs differed according to a lesion site, size, and macroscopic appearance. On endoscopic biopsy, FNR was related with early-stage KS  and site of esophagus, whereas SMT with ulceration is relatively easy to diagnose. Caution should be exercised when performing endoscopic biopsy of these lesions in AIDS patients and evaluating the histological features.


Tyrosine kinase inhibitors specific for BCR-ABL, were a major breakthrough in CML therapy. Second generation tyrosine kinase inhibitors  are indicated for imatinib resistant and intolerant patients. Present guidelines recommend continuous drug dosing for maintaining remission. There is no available data concerning the optimal duration of dasatinib therapy. We report the case of an imatinib intolerant patient who succeeded a complete molecular remission with dasatinib. Dasatinib was stopped bacause of intolerance, but complete molecular remission was sustained for one year and minor molecular remission for 27 months after discontinuation of dasatinib. Tyrosine kinase inhibitors specific for BCR-ABL, were a major breakthrough in CML therapy. Second generation tyrosine kinase inhibitors  are indicated for imatinib resistant and intolerant patients. Present guidelines recommend continuous drug dosing for maintaining remission. There is no available data concerning the optimal duration of dasatinib therapy. We report the case of an imatinib intolerant patient who succeeded a complete molecular remission with dasatinib. Dasatinib was stopped bacause of intolerance, but complete molecular remission was sustained for one year and minor molecular remission for 27 months after discontinuation of dasatinib.The introduction of tyrosine kinase inhibitors specific for BCR-ABL, were a major breakthrough in CML therapy . ApproxiSecond generation TKIs  are indicated for those patients who are refractory or intolerant to imatinib ,4. HowevBCR/ABL major fusion gene has increased gradually, but even now, 27 months after stopping dasatinib treatment, the patient is still in minor molecular remission , but complete molecular remission was lasting till one year after stopping dasatinib. The patient did not receive any kind of anti-CML therapy during this year. After one year without any TKI, the expression level of on Figure.Because of her several kind of comorbidities, her good quality of life, the actual hematological remission and also her reluctance to take any kind of TKIs, our therapeutical strategy is only watch and wait.This is the first report about maintaining complete molecular remission after one year of suspending dasatinib treatment.CML, Chronic myeloid leukemia; TKI, Tyrosine kinase inhibitor.The authors have no relevant conflict of interests.KG, DK and GD were responsible for the clinical guidance of the patient, Judit Csomor, András Matolcsy and Csaba Bödör carried out the molecular biological examinations. All authors participated in drafting and critically revising the manuscript. All authors read and approved the final manuscript.


Environmental Health Perspectives (EHP) established the Paper of the Year in 2008 (EHP Classic Paper of the Year is given to the most highly cited Research Article, Commentary, or Review Article over the preceding 60 months. We are proud to announce that the 2013 EHP Classic Paper of the Year is “Effects of Particulate Matter on Genomic DNA Methylation Content and iNOS Promoter Methylation” by Letizia Tarantini, Matteo Bonzini, Pietro Apostoli, Valeria Pegoraro, Valentina Bollati, Barbara Marinelli, Laura Cantone, Giovanna Rizzo, Lifang Hou, Joel Schwartz, Pier Alberto Bertazzi, and Andrea Baccarelli, published in the February 2009 issue of EHP. in 2008  as a wayThis research has had a significant influence on human environmental health research. The study by EHP congratulates Tarantini and colleagues for their sustained contribution to the environmental health science literature. They should be recognized for their outstanding contribution to the rapidly evolving field of environmental epigenetics.


Case. We present a case of an isolated bulbar conjunctival KS on a 43-year-old HIV positive male, with no other lesions. Excision and cryotherapy were performed, and the patient remains free of lesions to date. Conclusion. Isolated bulbar conjunctival KP is an unusual site for its initial presentation and must be kept in mind in HIV positive patients.Kaposi's sarcoma (KS) is a malignant vascular tumor, caused by the human herpesvirus 8. It is one of the commonest tumors in human immunodeficiency virus (HIV) patients and not uncommonly the first manifestation of acquired immunodeficiency syndrome (AIDS).  Ocular manifestations of acquired immunodeficiency syndrome (AIDS) are varied, affecting almost all structures of the eye. The posterior segment is mainly affected; however, less frequently ocular adnexa, anterior segment, orbit, and neuroophthalmic lesions can also occur. Human immunodeficiency virus- (HIV-) associated disease occurs in 50–90% of patients in the course of their illness .μ [Kaposi's sarcoma (KS) is the most common lesion of the anterior segment in these patients . It is aμ .KS is a malignant neoplasm of the vascular endothelium that can affect the eyelids, the lacrimal sac or gland, the orbit, and bulbar or palpebral conjunctiva. Isolated KS of the conjunctiva was first reported in 1967 . If it aWe report a clinical case of a patient with an isolated conjunctival KS and its treatment.A 43-year-old HIV positive male since August 2010 presented with a three-month history (since December 2010) of a growing mass in the bulbar conjunctiva of the right eye. The patient had previously undergone right eye cataract extraction with insertion of intraocular lens in 1992 in another medical institution.3.On ophthalmic examination, the best corrected visual acuity was 20/25 on his right eye. The biomicroscopy revealed two reddish nodular lesions localized in the superior and inferior nasal bulbar conjunctiva, loosely connected to the underlying tissue . FundoscAn excisional biopsy and cryotherapy on the conjunctival edges were done on January 2011. Histologic report of the biopsy was consistent with the diagnosis of KS with conjunctival edges free of neoplasm . CD31 anThe patient initiated standard antiretroviral therapy in January 2011 and remains in followup in the Infectious Diseases Department. Two years later the patient remains with no signs of recurrence .Improved survival as a result of antiretroviral therapy has led to an increase in systemic and ocular complications. KS is the most common tumor arising in HIV-infected patients. Ophthalmic presentation often occurs in the context of systemic KS, and isolated conjunctival lesions are rare –7.KS may affect many structures of the eye. Involvement of the conjunctiva appears as a flat, reddish lesion, most often localized in the inferior conjunctiva and fornix. It may be misdiagnosed with chronic subconjunctival haemorrhage.The differential diagnosis includes granuloma pyogenicum, cavernous haemangioma, foreign-body granuloma, malignant melanoma, metastatic tumor, and chronic subconjunctival haemorrhage .Dugel et al.  have claBiopsy with histopathological and immunohistochemistry report has an essential role in the diagnosis.surgical excision: in extensive, large tumors, loosely connected with healthy tissue;radiotherapy: for more extensive lesions;cryotheraphy: for early flat tumors;local injection of cytostatics , 10.The type of treatment depends on the size and location of the tumor. KS responds well to treatment byThis case describes an isolated conjunctival Kaposi's sarcoma as primary presentation of AIDS, which is an unusual location for KS .Complete surgical excision with cryotheraphy on conjunctival edges proved to be effective in our patient, with no evidence of clinical recurrence.This case emphasizes the importance of considering KS as a possible diagnosis in HIV positive patients. Also, KS can be the first AIDS-defining illness in patients with decreased immunological response.


Lipid bilayer membranes are among the most ubiquitous structures in the living world, with intricate structural features and a multitude of biological functions. It is attractive to recreate these structures in the laboratory, as this allows mimicking and studying the properties of biomembranes and their constituents, and to specifically exploit the intrinsic two-dimensional fluidity. Even though diverse strategies for membrane fabrication have been reported, the development of related applications and technologies has been hindered by the unavailability of both versatile and simple methods. Here we report a rapid prototyping technology for two-dimensional fluidic devices, based on in-situ generated circuits of phospholipid films. In this “lab on a molecularly thin membrane”, various chemical and physical operations, such as writing, erasing, functionalization, and molecular transport, can be applied to user-defined regions of a membrane circuit. This concept is an enabling technology for research on molecular membranes and their technological use. Supported molecular phospholipid films are versatile model membrane architectures23578910111516172023Writing  Preparation of stock vesicle suspensions, (II) Preparation of small unilamellar vesicles. De-ionized water from a Milli-Q system (Millipore) was used for all preparations.3PO4 (Sigma Aldrich), 30 mM KH2PO4 (Sigma Aldrich), 3 mM MgSO4*7H2O (Merck) and 0.5 mM Na2EDTA (Sigma Aldrich). The pH was adjusted to 7.4 with H3PO4 (Sigma Aldrich). The rehydrated lipid cake was placed in the fridge (4°C) overnight. In the final step the lipid cake was sonicated at 120 W/35 kHz  at room temperature for 15–30 s, to induce the formation of giant vesicles of varying, mainly multiple lamellarity.For each recipe, a designated amount (see table) of lipids and lipid conjugates in chloroform were mixed and diluted with chloroform to a total concentration of 10 mg/ml. 300 μl of this solution was placed in a 10 ml round bottom flask, and the chloroform was removed in a rotary evaporator at reduced pressure (−80 kPa) over a period of 6 hours. The dry lipid film at the walls of the flask was rehydrated with 3 ml of PBS buffer containing 5 mM Trisma Base (Sigma Aldrich), 30 mM K2EDTA (Sigma Aldrich), adjusted to pH = 7.4 and sonicated using a Sonics & Materials Vibra Cell™ High Intensity Ultrasonic Liquid Processor  at 15°C for 10 minutes. The sonicated samples were subsequently ultra-centrifuged at 40,000 rpm at 15°C for 30 minutes to separate multilamellar aggregates and tip debris . The small unilamellar vesicles in the supernatant were transferred to a separate tube. Critical: For high quality membrane fabrication, the vesicle suspensions must be freshly prepared. Upon prolonged storage, vesicle preparations form agglomerates.Small unilamellar vesicles were prepared on the day the experiments were conducted. 100 μl of the desired vesicle stock solutions were diluted (1:10) with TRIS buffer [125 mM NaCl (Sigma Aldrich), 10 mM TRIS (VWR), 1 mM Nai.e., the extent of the region to which solutions are delivered.The original multifunctional pipette with three on-chip solutions has been reported elsewhereThe preparation of plain and patterned glass surfaces is described in A.A. proposed the concept. A.A., I.G. and A.J. designed the experiments. A.A., I.G. and B.H. performed the experiments. A.A. analyzed data and performed simulations. A.A. prepared figures. A.J., A.A. and I.G. wrote the manuscript.Supplementary InformationVideo 1. Deposition.Video 2. Writing and erasing.Video 3. Functionalization.Video 4. Other examples.


Antibody-mediated rejection (AMR) is an important cause of graft loss after organ transplantation. It is caused by anti-donor-specific antibodies especially anti-HLA antibodies. C4d had been regarded as a diagnosis marker for AMR. Although most early AMR episodes can be successfully controlled or reversed, late and chronic AMR remains the leading cause of late graft loss. The strategies which work in early AMR have limited effect on late/chronic episodes. Here, we reviewed the lines of evidence that late/chronic AMR is the leading cause of late graft loss, characteristics of late AMR, and current strategies in managing late/chronic AMR. More effort should be put on the management of late/chronic AMR to make a better long term graft survival. Organ transplantation now represents the treatment of choice for patients developing end stage organ failure . HoweverAMR is caused by anti-donor-specific antibodies, mostly anti-HLA antibodies , 9. SomeObviously, a new marker for AMR is extremely necessary. It is reported that microcirculating inflammation is strongly correlated with alloantibody reactivity; however, whether it is can be used as maker of AMR is still of contradictory . T-box eThe importance of CAMR is increasingly recognized. It has been known as a major cause of late graft dysfunction in renal transplantation. Banff 07 consensus conference  describeAMR episodes occurring at different time periods seem to be different clinical subentities –21. TheyA eliminated term, chronic allograft nephropathy, had been regarded as a main cause of late graft loss . HoweverFurther prospective study from the same group  investigThe development of chronic AMR, especially TG usually takes years, and there still lacks of a perfect animal model to mimic the lesions during chronic AMR; thus, the natural history of chronic AMR is still not clear. Recently Wiebe et al.  have monA recent population based study  revealedChronic AMR is a B-cell-mediated production of immunoglobulin (Ig) G antibody against a transplanted organ. Based on this pathophysiologic condition, rituximab, IVIG, and bortezomib have been used as treatment for chronic AMR recently. However, till now, there is no standardized treatment for late/chronic AMR. The strategies that can effectively reverse early AMR do not work as well in late episodes; thus, an ounce of prevention is really worth a pound of cure . As late2 body surface area) 1 week after the last IVIG infusion. Four of the six patients had good response to this treatment; they had improved or stabilized eGFR. Further prospective studies from the same group showed that under this treatment, in the means of eGFR, 70% patients responded to treatment as measured 6 months after intervention, and this response persisted over a 24-month observation period. The rationale for the rituximab/IVIG treatment was to use IVIG for its immunomodulatory action and then rituximab for prevention of further antibody production.Several single center studies showed that the combination treatment with rituximab/IVIG may be a useful strategy for the treatment of chronic AMR. In 2008, a German group  publishe2 once on day 1), whereas IVIG (0.4 g/kg once daily on day 2 to 5) was given only to 3 patients. About the treatment safety, 3 out of 4 patients underwent therapy with rituximab/IVIG without side effects. One patient had severe, possibly rituximab-associated lung toxicity. Their study showed that rituximab/IVIG may be a useful strategy for the treatment of chronic AMR. Another pilot study showed that rituximab/IVIG treatment took effect in 3 out of 4 patients. Early stage of chronic AMR has better response than advanced stage [At the year 2009, Fehr et al.  publisheed stage . Anyway,ed stage . A retroed stage  studied Bortezomib is a proteosome inhibitor that leads in vitro to apoptosis of alloantibody-producing plasma cells . It has As complement plays an important role in the pathogenesis of AMR, complement-blocking agents could be used for the treatment of AMR. Eculizumab is a humanized monoclonal antibody against complement C5. It can bind to the C5 protein and inhibit conversion of C5 to C5b, thus preventing formation of the membrane attack complex (C5b–9). Eculizumab has been used to rescue atypical hemolytic uremic syndrome after renal transplantation . A prospThe spleen acts as a repository for memory B cells and plasma cells; thus, splenectomy is supposed to be effective in treating AMR. There is data suggesting that splenectomy alone can lead to rapid diuresis and immediate restoration of renal function . Rescue The liver allograft is generally regarded as relatively resistant to AMR. The resistance is attributed to a variety of characteristic features of liver which contribute to the clearing and dilution of antibodies or antigen-antibody complexes, such as Kupffer cell phagocytosis, large sinusoidal surface area, dual afferent hepatic blood supply, and secretion of soluble MHC class I antigen . For manAMR is also involved in other organ transplantation, especially for heart transplantation. DSA binding to the heart allograft causes myocardial injury predominantly through immune complex activation of the classical pathway of the complement cascade , and thuBesides renal and heart transplantation, AMR is also a major complication causing graft injury after lung , pancrea


MoSGrid is the acronym for Molecular Simulation Grid, a BMBF funded joint research project with the aim to offer grid services for the broad field of molecular simulations in the D-Grid infrastructure. Besides tendering various codes ranging from quantum molecular calculations  via molecular dynamics (e.g. Gromacs) to docking approaches (e.g. FlexX) for high performance computing, one of the main goals is the integration of metadata annotation for data mining and knowledge generation.http://www.mosgrid.de), which will offer intuitive access to various tools and will support the users with workflows, for an easy import of molecular data, a simple setup and submission of calculations as well as extraction of relevant results. The portal will hide the complexity of the underlying technology by providing a unified user interface making computational chemistry in general more readily available.Molecular simulation codes and computational resources are accessed via the MoSGrid portal (MoSGrid’s server-based portal is available as open-access and open-source software. Users are relieved from software installations and do not need to have knowledge about the underlying infrastructure. The portal includes portlets specifically set up for the various simulation programs. Commonly used workflows, simple or complex, can be stored in recipe repositories and are available for every user. Moreover, users can develop, improve, publish, and use workflows for their everyday tasks.


Signaling involving dopaminergic neurons in the brain is important for determining the saliency of sensory input, the urgency of sensory import, the actual value of an experience as compared to its expected value, and the process of converting motivation to action. Drugs that are antagonists for the dopamine D2 receptor subtype are clinically effective in treating schizophrenia and a variety of other mental conditions. These drugs have a substantial effect on the levels and sub-cellular distribution of dopamine and its major metabolite DOPAC. Currently, there is no quantitative analysis of how these various changes can be accomplished. The goal of this work is to explain how the changes in dopamine and DOPAC levels and subcellular location elicited by the prototypical antipsychotic drug haloperidol occur.A computational model of a dopaminergic striatal varicosity was used. In the model, dopamine can be located in cytosol, vesicles, extracellular compartment, and a DOPAC-synthesis-secretory compartment. DOPAC can be located in cytosol, extracellular, and DOPAC-synthesis-secretory compartment. The model has two distinct sites for dopamine and DOPAC synthesis. The cytosolic site synthesizes dopamine for use in the signaling pool and metabolizes a portion of dopamine that is transiently located in cytosol. The DOPAC-synthesis-secretory complex site is hypothesized to be located on the plasma membrane and to be dedicated to the synthesis of DOPAC to be secreted into the extracellular compartment. The model also has enzymes involved in dopamine and DOPAC synthesis, transporters that move dopamine between compartments, and exocytosis.Haloperidol induces a small increase in extracellular dopamine, a somewhat larger increase in extracellular DOPAC, a larger increase in tissue DOPAC, a large increase in rate of dopamine synthesis, and no change in level of dopamine. The increase in extracellular dopamine was simulated by increasing rate of secretion of dopamine into extracellular space by exocytosis. An increase in synthesis of signaling dopamine was required to maintain the elevated levels of extracellular dopamine. The increase in extracellular DOPAC was simulated by increasing rate of dopamine synthesis in the DOPAC-synthesis-secretory compartment. With only these manipulations, the model failed to match experimental data in terms of increase in total tissue DOPAC and increase in the rate of dopamine synthesis. These discrepancies were resolved by increasing the passive diffusion of dopamine from vesicles to cytosol and further increasing the rate of dopamine synthesis for the signaling pool. Literature data indicate that haloperidol does accumulate in and partially alkalinize amine neurotransmitter storage vesicles, which would increase passive diffusion of dopamine from the vesicles.I conclude that the increases in extracellular dopamine and DOPAC are likely mediated by antagonism of dopamine D2 receptors but that the large increase in tissue DOPAC and much of the increase in dopamine synthesis result from physico-chemical properties of haloperidol rather than block of dopamine D2 receptors. Furthermore, the results suggest dopaminergic varicosities possess a mechanism for monitoring amount of stored dopamine and modulating dopamine synthesis to maintain constant stores.


I–V converter in order to measure ultralow conductances of molecular rods trapped between two gold leads. The current resolution of the setup reaches down to 10 fA. We report single-molecule conductance measurements of an anthracene-based linearly conjugated molecule (AC), of an anthraquinone-based cross-conjugated molecule (AQ), and of a dihydroanthracene-based molecule (AH) with a broken conjugation. The quantitative analysis of complementary current–distance and current–voltage measurements revealed details of the influence of π-conjugation on the single-molecule conductance.π-Conjugation plays an important role in charge transport through single molecular junctions. We describe in this paper the construction of a mechanically controlled break-junction setup (MCBJ) equipped with a highly sensitive log  Molecular electronics has expanded tremendously during the past ten years –13. A co10Charge-transport characteristics of single molecules or of a few molecules trapped between two leads were studied in various experimental platforms. These include scanning tunneling microscopy (STM) –29, curr16STM-BJ and MCBJ are the two most popular and reliable approaches for single-molecule conductance measurements. Reed et al. , KerguerAC), an anthraquinone-based cross-conjugated wire (AQ), and a dihydroanthracene-based wire with a broken π-conjugation (AH) (In the present paper we explore the influence of π-conjugation on the conductance of single-molecule junctions of oligophenylene ethynylene (OPE)-type molecules contacted to gold leads. We have chosen three rigid dithiolated molecular wires with different conjugation patterns: An anthracene-based linearly conjugated wire (ion (AH) .I–V converter /∆z = 5.5 nm−1) of the STM-BJ experiments was chosen to scale the traces acquired in the MCBJ setup.The distance between the two gold electrodes in the MCBJ setup is calibrated with the assumption that the tunneling decay is identical to that in a STM-BJ setup under the same experimental conditions. Conductance–distance traces representing a well-defined tunneling response, e.g., without molecular plateaus, were recorded in a STM-BJ and in a MCBJ configuration. Subsequently the decay constant , which were cleaned in boiling 25% nitric acid and Milli-Q water, and dried in a stream of argon. A gold wire of 100 μm in diameter was subsequently fixed on these sheets with two drops of preheated epoxy . The distance between the two drops of epoxy glue was adjusted to be less than 500 μm. Next, the sample was conditioned overnight at 60 °C for epoxy polymerization. The freely suspended part of the wire was notched with a scalpel blade under an optical microscope to fabricate a constriction point. The as-prepared sample sheets were cleaned in boiling Milli-Q water for 15 min, rinsed with isopropanol and dried with argon before each experiment.The Kel-F liquid cell including its cover, Kalrez O-ring and Teflon tubes for argon purging and solution exchange were cleaned in three alternating boiling cycles in 25% nitric acid and Milli-Q water to remove absorbed contaminants.The sample sheet was first mounted on the sample holder of the MCBJ setup. Subsequently, the liquid cell was installed on top of the sample with a Kalrez O-ring attached to prevent leakage of the solution. The closed liquid cell was flushed with argon through an inert-gas cycling system to remove oxygen, and then the solution containing the test molecule was pumped into the liquid cell through a triple valve. The last step was repeated three times to reduce contaminations. Subsequently, the input and output valves for solution exchange and gas purging were closed, and the experiment started.Basic principles of the STM-BJ experiment, data analysis and sample preparation were described previously ,33.AQ followed a method reported previously [AC and of the molecular wire with broken symmetry AH will be communicated elsewhere [AC indicated that a high yield of single-molecule junctions is obtained in the absence as well as in the presence of in situ deprotecting agents, such as tetrabutyl ammonium hydroxide or triethylamine. As a consequence, and to keep the number of different species in the sample solution to a minimum [The synthesis of the antraquinone-based cross-conjugated wire eviously . Detailslsewhere ,56. The  minimum , we perf−1 as the typical rate for the movement of the pushing rod in the breaking process. This value translates into an approximate lateral movement between the two gold leads of about 1 nm. All data shown in the following sections and used for the analysis represent opening traces, which were recorded after breaking a gold–gold contact. AC. All curves start with characteristic steps and plateaus in the region between 10 and 1 Go (orange part), representing the breaking of gold–gold atomic contacts. The last step is observed around 1 G0. After the gold–gold monatomic contact is broken, the two “separated” gold electrodes snap back and a nanogap is created with typical conductances ranging between 10−2G0 and 10−4G0. The snap-back process is too fast to be recorded with better resolution. At lower conductances we observed two distinctly different types of traces, those without molecular plateaus  and those with molecular plateaus  in the range of 10−4 to 10−5G0. The noise level is reached below 10−8.2G0, which provides a wide window of over eight orders of magnitude for the single-molecule conductance measurements.The measurements of conductance–distance traces in the MCBJ set up were carried out with 50 nm·sAC|gold molecular junctions with a characteristic plateau. The three representative individual traces reveal a single plateau conductance at around 10−4.5G0, which indicates the formation of a single-molecule junction between the two electrodes. The current noise is attributed to the thermal vibration of the molecular junction at room temperature. The conductance traces exhibit an abrupt decrease upon breaking of the molecular junction until the noise level is reached. The most probable conductance of the molecular junction was obtained by statistical analysis of the data. The resulting histogram, as constructed from the three red traces, is plotted in the right panel of −4.5G0, which is equal to 2.5 nS, the most probable single-molecular junction conductance of AC from a limited data set of three individual traces. . Applying the same analysis method to the blue traces did not lead to any clear feature between 10−1G0 and 10−8G0, which supports the assignment of the two types of traces.The blue traces in I–V measurements we controlled the opening and closing cycles by slowly moving the pushing rod at a rate of 0.5 nm·s−1 and we swept simultaneously the bias voltage Vbias from −0.4 V to +0.4 V at a rate of 25 V·s−1 at various positions. This approach resulted in a set of I–V curves spanning a range of conductance during a single opening and closing cycle, which correspond variously to the Au–Au contacts  versus distance traces are linear until the noise level is reached. The second type of curves . This value represents the most probable conductance of a gold|AC|gold single-molecule junction, and is in perfect agreement with the result of the continuous current–distance measurements  and Γ = 0.0012 eV. The negative sign is concluded from the following: Considering the Fermi energy of gold as −5.0 eV, we estimated −5.53 eV as the position of the EHOMO level. This result is in good agreement with AM1-RHF calculations, performed with Hyperchem Release 7.52, and the level alignment, based on UPS data, of a related AQ-type molecular wire, EHOMO(Hyperchemcorr) = −5.74 eV [2Cl2 [AQ-type molecular junctions is HOMO-dominated.Fitting the model, as represented by −5.74 eV . The HOMeV [2Cl2 . Based oThe coupling parameter Γ appears to be rather small as compared to those for other dithiole-terminated molecular wires attached to gold leads ,18. ThesAC, constructed from the above I–V data  could not be monitored reliably due to the relatively slow response of the log I–V converter in the pA range. The I–V converter could not follow precisely enough the current change in the low-conductance range upon sweeping the bias voltage at a rate of 25 V·s−1.The conductance histogram based on the statistical analysis of AC, AQ, AH and, for comparison, also the target-molecule-free THF/decane solution, as obtained in a series of MCBJ measurements. All experiments were carried out under identical conditions and analyzed with the strategies introduced above. We note that the histograms constructed for the blank control experiment  in the conductance histogram of the cross-conjugated anthraquinone wire AQ. This feature is well separated from the noise level, which is located at 10−8.8G0. The junction conductance of AQ is approximately 300 times lower than that of AC. This trend demonstrates that the cross-conjugated motif of AQ indeed gives rise to a lower conductance as compared to the linear-conjugated AC, which is in agreement with ab initio transport calculations predicting a destructive quantum interference present in AQ, but which is absent in AC molecular bridges [AQ molecular junction also illustrates the high sensitivity of our new MCBJ setup. bridges . From a AH with a broken π-conjugation. The plot reveals one main feature at 10−6.3G0 (39 pS) and a faint second feature around 10−4.5G0 (2.5 nS), the latter being 5 times larger and close to the data reported for AC.G0 to define the relative zero of the distance scale [AC in −4.5G0 and a second, less dense patch of data points, around 10−7.2G0 indicating a low conductance feature. The 2-D histogram of AQ reveals only one clear molecular feature, which is found around 10−7.0G0 . We extracted the most probable stretching distance of breaking for the high conductance plateau of AC in the range of 10−1G/G0 to 10−6 G/G0 to explore further details of the low-conductance state. As illustrated in AC molecular junction breaks is 2.5 nm, while the most probable stretching distance up to the end of the high-conductance molecular plateau is obtained as 2.3 nm (inset in AC|gold junctions is estimated at 3 nm by adding the “snap-back” distance of 0.65 nm [AC (2.7 nm). We propose that the strong gold–thiole bond leads to the “pulling-out” of surface gold atoms just before the breaking of the molecular junction. The low-conductance feature is attributed to π-stacking interactions between two molecules attached only at one end of the leads [We further analyze the stretching distance from the breaking of gold–gold contacts until the noise level is reached  upon breaking of an atomic gold–gold contact  measurement"), and not for AQ and AH. The rather low junction-conductance characteristics of these two molecules as recorded simultaneously during single stretching traces were too much distorted by the onset of instrumental noise.junction ,56. CompAQ and AH are smaller than the molecular length indicating that the junctions breaks more frequently before the molecule assumes an extended atop–atop configuration between the two ends of the gold leads. A peculiarity appears in the form of a weak conductance feature observed for AH around 10−4.5G0  was also observed in current-probe atomic force microscopy (CP–AFM) [Finally, we notice that the same sequence of conductance values as found in the MCBJ-experiments ((CP–AFM)  and EGaI(CP–AFM)  of theseI–V converter to measure ultralow conductances of molecular rods formed between two gold leads. In particular, we carried out quantitative single-molecule conductance experiments on linearly conjugated, cross-conjugated, and broken-conjugated examples of dithiolated molecules of the OPE family. The current sensitivity of the setup reaches down to 10 fA. Our experiments demonstrate that the conductance of the linearly conjugated molecule AC is several hundred times higher than that of the broken π-conjugated molecule AH, and the conductance of AH is about five times higher than that of the cross-conjugated molecule AQ. The latter result is attributed to destructive quantum interference present in the AQ molecular bridge [AC appears to be capable of forming a large number of fully extended gold|molecule|gold junctions. The other two molecules AQ and AH form junctions that break before reaching full extension.We described in this paper the construction of a mechanically controlled break-junction setup (MCBJ) equipped with a highly sensitive log r bridge . All ditThese experimentally observed trends in the values of the single-molecule conductances as well as in the stability of the respective molecular junctions reveal the key role of π-conjugation in the charge transport through rigid-rod OPE-type single-molecule junctions. Moreover, the good agreement between the different measurement approaches employed in this paper (current–distance and current–voltage traces from MCBJ and STM-BJ) confirm the reliability of our measurements. The observation of similar trends in the main conductance values discovered in single-molecule  and parallel-molecule junction experiments  confirms the complementarity of the various experimental platforms, in both their similarities as well as their distinct differences.


The MoSGrid (Molecular Simulation Grid) project is currently establishing a platform that aims to be used by both experienced and inexperienced researchers to submit molecular simulation calculations, monitor their progress, and retrieve the results. It provides a web-based portal to easily set up, run, and evaluate molecular simulations carried out on D-Grid resources. The range of applications available encompasses quantum chemistry, molecular dynamics, and protein-ligand docking codes.In addition, data repositories were developed, which contain the results of calculations as well as “recipes” or workflows. These can be used, improved, and distributed by the users. A distributed high-throughput file system allows efficient access to large amounts of data in the repositories. For storing both the input and output of the calculations, we have developed MSML (Molecular Simulation Markup Language), a CML derivative . MSML has been designed to store structural information on small as well as large molecules and results from various molecular simulation tools and docking tools. It ensures interoperability of different tools through a consistent data representation.http://www.mosgrid.de the new platform is already available to the scientific community in a beta test phase. Currently, portlets for generic workflows, Gaussian, and Gromacs applications are publicly accessible [At cessible ,2.


E. coli AGEs are continually formed as high-molecular weight protein complexes. Moreover, we showed that AGEs are removed from the cells by an active, ATP-dependent secretion and that these secreted molecules have low molecular weight. Taken together, these results indicate that E. coli contains a fraction of low molecular weight AGEs, in addition to the high-molecular weight AGEs. Here we show that the low-molecular weight AGEs originate from high-molecular weight AGEs by proteolytic degradation. Results of in-vitro and in vivo experiments indicated that this degradation is carried out not by the major ATP-dependent proteases that are responsible for the main part of bacterial protein quality control but by an alternative metal-dependent proteolysis. This proteolytic reaction is essential for the further secretion of AGEs from the cells. As the biochemical reactions involving AGEs are not yet understood, the implication of a metalloprotease in breakdown of high molecular weight AGEs and their secretion constitutes an important step in the understanding of AGEs metabolism. Advanced Glycation End Products (AGEs) are the final products of non-enzymatic protein glycation that results in loss of protein structure and function. We have previously shown that in  Advanced Glycation End-products (AGEs) are the final products of non-enzymatic glycation formed by the reaction of reactive carbonyls (e.g.- reducing sugars) with primary amine-containing amino acids of proteins. One of the first steps in this glycation process is the formation of Amadori-modified proteins (AMPs) which are reversible intermediates. These AMPs can further developed, in an oxidation-dependent manner, to form advanced protein complexes, that contain irreversible, highly stable high molecular weight AGEs –4 . Alth In mammals, AGEs were shown to accumulate both intracellularly and extracellularly with age and to participate in the pathophysiology of several age-related diseases such as cardiovascular disease, Alzheimer’s disease and complications of diabetes mellitus –10. Theyin-vivo and in-vitro but did not affect degradation of AGEs, suggesting that AGEs are not degraded by the cellular proteasome . In. In22]. in-vitro and in-vivo, that metallo-proteases are involved in AGEs degradation. We also show that AGEs degradation is a required step for AGEs secretion, as inhibition the degradation results in reduced secretion and accumulation of AGEs inside the bacterial cell. These findings further our understanding of AGEs metabolism and can shed a light on the general mechanisms responsible for AGEs degradation in bacteria and possibly other organismsIn this work, we investigated the proteolytic stage that precede AGEs secretion and show, both in-vivo we arrested protein translation and AGEs secretion, using chloramphenicol and arsenate [The finding that AGEs – formed as high molecular weight compounds - are also found in low-molecular weight fractions  leads toarsenate , and meain-vitro assay in which we determined the kinetics of AGEs degradation under several experimental conditions. High-molecular-weight protein fractions were incubated at 37°C, fractionated, and the levels of high and low molecular-weight AGEs were measured. After 3 hours of incubation the majority (~70%) of AGEs were proteolytically cleaved to low molecular-weight compounds  are irreversible toxic molecules that accumulate in cells with time. Given their strong correlation to various diseases, the physiological effects of AGEs were extensively studied in human and conditions that affect the kinetics of their formation are well documented. However, pathways that participate in AGEs metabolism are poorly characterized and mechanisms that reduce their toxicity have not been extensively studied.In mammals, it was found that AGEs are secreted from the body by the urinary system and that ineffective clearance of AGEs leads to their accumulation ,22. WhilE. coli cells and provided evidences that inside the bacterial cells high-molecular-weight AGEs are degraded to form low-molecular weight compounds. This degradation is attributed to the protein moiety of the AGEs as the total fluorescence signal of the AGEs remains the same while their size distribution changes with time. To our best knowledge, this is the first direct evidence for intracellular degradation of AGEs. Non-enzymatic glycation is a ubiquitous process, and therefore we have suggested that bacteria can constitute a powerful tool in the study of this complicated pathway. Here we showed that AGEs are proteolyticlly degraded inside In-vitro assays suggest that AGEs degradation is enzymatic, since denaturative agents such as urea and proteases inhibitor cocktail have reduced the rate of cleavage. Further examination on the various components of the proteases inhibitor cocktail revealed that metallo-proteases inhibitors are the most effective in prevention of AGEs degradation suggesting that metallo-proteases are the main players responsible for AGEs degradation. Similar observations were made in-vivo. These results indicate that the major ATP-dependent proteases, responsible for the main part of bacterial protein quality control, are not involved in the degradation of AGEs. Supporting this conclusion is the fact that Arsenate, a glycolysis uncoupler that severely reduces intracellular ATP concentration, did not affect the kinetics of AGEs degradation inside the bacterial cell. These results are compatible with the data obtained in eukaryotes suggesting that the proteasome, the eukaryotic equivalent to bacterial ATP-dependent proteases, is not responsible for proteolytic degradation of AGEs in higher organisms ,16.E. coli, there are 30 metallo-proteases of which 5 are essential proteins. In an attempt to characterize the specific proteases that are involved in AGEs degradation we used mutants from the Keio knockout library collection [pqql deletion strain, lacking a putative zinc peptidase [In llection , and anaeptidase , which aE. coli, this degradation appears to be independent of energy, although the subsequent secretion of the low molecular weight AGEs is carried out by the energy-dependent efflux-pumps system [in-vivo the degradation and secretion are linked, but have shown that the degradation is required for the secretion. Thus, we propose that AGEs degradation is a preliminary step, necessary for the clearance of toxic AGEs.We propose the following model; inside bacterial cells AGEs are mainly formed as high-molecular-weight modified proteins. Since AGEs are irreversible molecules they accumulate with time inside the cells. Clearance of these AGEs by secreting them is one strategy of the cells to cope with AGEs accumulation. However, as AGEs are not secreted as high molecular weight compounds, the protein moieties have to be degraded inside the cells prior to secretion. This degradation is probably performed by one or more metalloproteases. Unlike the major degradation pathways in s system . At thisUnlike other cellular damage, accumulation of AGEs presents an additional challenge to living organisms as AGEs can further react with other macromolecules such as lipids, proteins, and DNA leading to extensive damage potential . On the E. coli K12 wild type strain MG1655 was used throughout. Cultures were grown with aeration at 37°C in standard LB medium (Difco). When required cultures were treated with 100 µM / 1 mM 1,10 phenantholine (Sigma), 100 µg/ml chloramphenicol (Sigma) or 50 µg/ml arsenate (Sigma). The in-vivo, was measured shortly after the inhibitor was added, in order to minimize a bias caused by its effect on bacterial growth rate. The accumulation of AGEs shown in the first 10 minutes is followed by a reduction of total AGEs due to growth arrestThe effect of 1,10 phenantholine on AGEs levels Determinations of the levels of secreted AGEs were performed in a non-growing culture using carbon-free media, as previously described .Lysates were obtained using the Qproteome-bacterial Protein Prep kit (QIAGEN) according to the manufacturer's recommendation. The lysates were further separated into purified proteins and low molecular weight compounds using GE Healthcare HiTrap desalting columns, equilibrated with PBS, using Amersham AKTA prime plus FPLC system. These columns possess exclusion limit of 5000 g/mol of globular compound, thus compounds with lower molar mass are hereafter referred to as low molecular weight compounds. Nucleic acids were digested as part of Qproteome extraction procedure, and therefore were present mostly in the low molecular weight fraction. For AGEs size exclusion filtration measurements  samples AGEs were quantified using the natural AGE-specific fluorescence by scanning emission ranging from 400 nm to 500 nm upon excitation at 370 nm at 37°C, in a HORIBA scientific FluoroLog-3 Spectrofluorometer. Data represent the 440 nm emission peak


Mycobacterium tuberculosis, also provides a survival niche from which the bacteria may disseminate. The tuberculosis lesion is highly dynamic and shaped by both, immune response elements and the pathogen. In the granuloma, M. tuberculosis switches to a non-replicating but energy-generating life style whose detailed molecular characterization can identify novel targets for chemotherapy. To secure transmission to a new host, M. tuberculosis has evolved to drive T cell immunity to the point that necrotizing granulomas leak into bronchial cavities to facilitate expectoration of bacilli. From an evolutionary perspective it is therefore questionable whether vaccination and immunity enhancing strategies that merely mimic the natural immune response directed against M. tuberculosis infection can overcome pulmonary tuberculosis in the adult population. Juxtaposition of molecular pathology and immunology with microbial physiology and the use of novel imaging approaches afford an integrative view of the granuloma’s contribution to the life cycle of M. tuberculosis. This review revisits the different input of innate and adaptive immunity in granuloma biogenesis, with a focus on the co-evolutionary forces that redirect immune responses also to the benefit of the pathogen, i.e., its survival, propagation, and transmission.A granuloma is defined as an inflammatory mononuclear cell infiltrate that, while capable of limiting growth of  Mycobacterium tuberculosis (M. tuberculosis), i.e., physiological and metabolic adaptation to the host environment, dynamics of replication, and synthesis and structures of the cell wall in order to define microbial factors important for virulence and persistence. On the other hand, infection immunologists analyzed innate and adaptive immune responses required to contain M. tuberculosis growth and dissemination, often welcoming the pathologist’s view on granuloma initiation, maintenance, and disintegration. In recent years, a more integrated view of tuberculosis pathogenesis has prevailed: the granuloma is not only recognized as a tissue reaction to limit bacillary growth and sequester infection but also as part of the successful life cycle of M. tuberculosis, thus representing the dynamic combat zone between both, the pathogen and host defense elements. This co-evolutionary perspective emphasizes the mutual shaping of the tissue microenvironment, which, at the same time, allows propagation and transmission of M. tuberculosis, yet restricts tissue damage to safeguard survival of the host. This review will highlight recent findings that have shifted the long-held paradigm that in TB the granuloma is primarily or uniquely relevant for protection of the host. This integrative model of the granuloma’s function in TB pathogenesis also challenges the concept that granulomas exclusively serve the interest of the pathogen.The pathogenesis of tuberculosis used to be the investigative domain of two relatively separate, albeit complementary disciplines. On the one hand, molecular microbiologists decoded the survival strategies of M. tuberculosis encounters is the alveolar macrophage, which phagocytoses but fails to kill the mycobacterial invaders, but does produce chemoattractants. Chemokines produced by alveolar macrophage and pneumocytes attract the first round of inflammatory cells, i.e., neutrophils, monocyte derived macrophages, NK cells, and γδ-T cells, which further promote inflammation and tissue remodeling , RpfB being the most relevant isoform out of five encoded in the M. tuberculosis genome based on persistence assays with targeted deletion mutants  allowing the mycobacteria to escape neutrophil-mediated killing. In contrast, an attenuated M. tuberculosis mutant lacking the virulence associated RD1 region fails to induce necrosis and falls victim to neutrophils’ armamentarium, i.e., ROI  were shown to drive granuloma necrosis in a model of mycobacteria-induced pulmonary immunopathology  do not efficiently kill mycobacteria unless activated by T cells  or during the latent phase (Rv2660c), not only boosts BCG-primed immunity but also induces multifunctional T cell-mediated immune protection on its own before and after exposure to M. tuberculosis  but is only a weak IFN-γ inducer by itself. It becomes mildly immunogenic only when fused to Ag85-ESAT6 thereby obviating immune-mediated exacerbation of the disease in infected individuals. In sum, the selection and combination of antigens and epitopes specific for different stages of TB may help outwit M. tuberculosis and control reactivation.Further considerations for improving vaccines take into account that It would be unreasonable to call the TB granuloma an unsuccessful host defence, as it successfully contains the infectious focus in more than 90% of cases. The 10% of individuals that progress toward TB disease suffer from a disbalanced inflammatory reaction, be it due to too little innate or adaptive immunity or due to unrestrained hypersensitivity reactions. There is no balance without a trade-off: in the case of TB this is the Janus face of T cell immunity . Any intervention thus must be regulatory in nature rather than proinflammatory, and the rational design of therapies and vaccines must take this into account.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.


Background. Transesophageal echocardiography (TEE) is used for the evaluation of the presence of left atrial appendage (LAA) thrombus prior to pulmonary vein isolation (PVI), while coronary computed tomography angiography (CCTA) is used for anatomic mapping during PVI. Methods. We compared the diagnostic performance of single phase CCTA to TEE in excluding the presence of LAA thrombus in patients undergoing PVI in 172 subjects performed during index hospitalization. Results. The mean age was 51 ± 13 years, a median CHADS2 score of 1  and a mean periprocedural INR of 2.1 ± 0.6. The prevalence of an LAA filling defect on single phase CCTA was 9.3% (6/183) and on TEE was 1.2% (2/183). Sensitivity, specificity, positive predictive value, and negative predictive value were 100% , 91.8% , 12.5% , and 91.8%  for the detection of LAA filling defect, respectively. Conclusion. Given the utility of a preprocedural single phase CCTA for the performance of PVI, the absence of a filling defect negates the need for a subsequent TEE as an adjunct for exclusion of LAA thrombus. Transesophageal echocardiography (TEE) historically is the gold standard for the evaluation of the presence of left atrial appendage thrombus in patients with atrial fibrillation. In a recent study, evaluating spontaneous echo contrast (SEC), left atrial (LA) appendage velocities, and aortic plaque TEE variables in the absence of definitive left atrial thrombus (LA) in five hundred and seventy-nine consecutive patients undergoing pulmonary vein isolation (PVI), all variables with the exception of LA thrombus were not predictive of the occurrence of stroke within thirty days on anticoagulation with Vitamin K Antagonist. . This isAdvances in coronary computed tomography angiography (CCTA) imaging have led to high spatial and temporal resolution. CCTA is emerging as a potential screening tool for exclusion of LAA thrombus . PatientA number of studies have assessed their application for determining the presence or absence of thrombus. Romero et al. completed a meta-analysis of detection of LAA thrombus by CCTA in patients with atrial fibrillation that demonstrated an overall high accuracy of CCT compared with TEE for the detection of LA/LAA thrombus. The mean sensitivity and specificity were 96% and 92% and positive predictive value and negative predictive value were 41% and 99% [The purpose of this review is to evaluate the diagnostic accuracy of single phase CCTA obtained routinely in our institution prior to pulmonary vein isolation (PVI) in comparison to TEE in excluding the presence of LAA thrombus and thus limiting the need for invasive procedures such as TEE to only patients with possible thrombus on CCTA.Chart review evaluating concomitant (within 48 hours) single phase 64- or 128-slice CCTA and TEE performed between January 2008 and June 2012 prior to PVI. All procedures were performed in symptomatic individuals with atrial fibrillation over the age of 18 at a single tertiary referral center . LAA was evaluated by both CCTA and TEE for filling defects. Imaging modalities were reviewed by a level II echocardiographer and a level III CT imaging staff to assess for the presence or absence of a thrombus.From January of 2008 to March of 2011, images were obtained using a retrospective helical protocol with a 64-slice CT scanner . From March of 2011 to March of 2012, studies were obtained utilizing a prospective sequential protocol with 60–80% image acquisition window. After March of 2012, 128-slice dual head scanner with a single heart beat image acquisition of the complete cardiac anatomy if a heart rate of less than 60 was achieved. . The subject's EE images were analyzed utilizing Merge Cardio  for the evaluation of the entire left atrial appendage. The subject's CCTA images were evaluated using Vitrea 15 software  .P values <0.05 considered significant.Statistical analysis was performed using IBM SPSS version 19.0 . Continuous variables are presented as means ± standard deviation and medians with interquartile range, as appropriate. Categorical variables are presented as frequencies with percentages. Comparison of means was performed using one-way ANOVA with post hoc Bonferroni correction, with 25,75 0,1 days). statement should read: Mean age was 51 ± 13 years with a median CHADS2 score of 1 , and a mean preprocedural INR of 2.1 ± 0.6. Prevalence of an LAA filling defect on single phase CCTA was 9.3% (16/172) versus TEE 1.2% (2/172). The diagnostic accuracy of single phase CCTA is 96.2%. Sensitivity, specificity, positive predictive value, and negative predictive value were 100% , 91.8% , 12.5% , and 91.8%  for the detection of LAA filling defect, respectively . CT imaging may identify a filling defect that if viewed by TEE will be assessed to be SEC. Shapiro et al. in their retrospective study detail this finding as a false positive. As stated by Shapiro et al., this issue may be addressed with a delayed scan protocol . Further2 score mainly between 0 and 1, similar population to ours, the complications rate reported postablation was <1% with zero incidence of death or stroke [In a prospective study of 500 consecutive AF ablation procedures performed on 424 patients with a mean age of 55 ± 11 years and a CHADSr stroke .Our findings not only confirms the findings of past studies illustrating the high sensitivity, specificity, and NPV between CCTA and TEE in excluding thrombus, but also confirms that a single phase CCTA if part of planned preparation prior to PVI is sufficient to exclude the presence of thrombus. CCTA should be used as an initial step to exclude the presence of filling defects and complimented by invasive procedures such as TEE for confirmation of an LAA thrombus or delayed phase CCTA in the appropriate clinical setting.


Aminochrome has been proposed to play an essential role in the degeneration of dopaminergic neurons containing neuromelanin by inducing mitochondrial dysfunction, oxidative stress, the formation of neurotoxic α-synuclein protofibrils, and impaired protein degradation. Here, we discuss the relationship between the oxidation of dopamine to aminochrome, the precursor of neuromelanin, autophagy dysfunction in dopaminergic neurons containing neuromelanin, and the role of dopamine oxidation to aminochrome in autophagy dysfunction in dopaminergic neurons. Aminochrome induces the following: (i) the formation of α-synuclein protofibrils that inactivate chaperone-mediated autophagy; (ii) the formation of adducts with α- and β-tubulin, which induce the aggregation of the microtubules required for the fusion of autophagy vacuoles and lysosomes.The molecular mechanisms involved in the neurodegenerative process of Parkinson's disease remain unclear. Currently, there is a general agreement that mitochondrial dysfunction,  VMAT-2 uses this proton gradient to take up one molecule of dopamine with the concomitant release of two protons [Dopamine is a neurotransmitter that plays an essential role in the control of movements and loss of dopaminergic neurons containing neuromelanin in the nigrostriatal system. In addition, dopamine is involved in the development of motor symptoms experienced in patients diagnosed with Parkinson's disease (PD). Dopamine is synthesized in a sequential reaction in which the cytosolic enzymes tyrosine hydroxylase (TH) and aromatic amino acid decarboxylase (AADC) catalyze the hydroxylation of the amino acid tyrosine to L-dihydroxyphenylanaline (L-dopa) and decarboxylation of L-dopa to dopamine, respectively. The protons of the hydroxyl group in dopamine dissociate when dopamine is localized in the cytosol at physiological pH. However, these protons are tightly bound to the hydroxyl group once dopamine is inside monoaminergic synaptic vesicles, which have a relatively low pH. The membrane of monoaminergic synaptic vesicles contains a vesicular monoaminergic transporter-2 (VMAT-2) that catalyzes the uptake of dopamine into these vesicles. These monoaminergic synaptic vesicles contain an ATPase that hydrolyzes ATP to ADP and Pi, and one proton , which is subsequently oxidized to dopamine o-quinone (reaction 2) by reducing two molecules of oxygen to superoxide radicals. The dopamine o-semiquinone radical does not strongly react with oxygen, leading to the formation of leukoaminochrome o-semiquinone radical during a one-electron reduction of aminochrome [o-semiquinone radicals can disproportionate, generating one molecule of dopamine o-quinone and one molecule of dopamine (reaction 3). A two-electron oxidation of dopamine to dopamine o-quinone is catalyzed by the enzyme tyrosinase. Notably, the presence of dopamine o-semiquinone is not detected by electron spin resonance [o-quinone is not stable in the cytosol at physiological pH and its amino chain cyclizes (reaction 5), generating aminochrome . These proteins are inactivated by aminochrome because dopamine o-quinone cyclizes immediately at physiological pH [The oxidation of dopamine can also be catalyzed by enzymes with peroxidase activity, such as prostaglandin H synthase, cytochrome P450 forms, dopamine  oxidase –20. Lactesonance . Howeveresonance , 21–24. esonance . Dopamingical pH .Aminochrome is the precursor to neuromelanin because neuromelanin formation is dependent on the rearrangement of aminochrome to 5,6-dihydroxyindole, which is then oxidized to 5,6-indolequinone followed by further polymerization to form neuromelanin  Figure . Postmorα-synuclein [α-synuclein protofibers is dependent on a specific point mutation [α-synuclein protofibrils. Aminochrome is also able to form adducts with mitochondrial complexes I and III, as well as isocitrate dehydrogenase [ Aminochrome forms adducts with proteins such as ynuclein , stabiliynuclein . In famimutation . Howeverrogenase , suggestrogenase , which hrogenase .α- and β-tubulin. Other studies also report the formation of aminochrome adducts with actin and β-tubulin [Aminochrome has been reported to disrupt the architecture of the cytoskeleton in cell cultures  by formi-tubulin . In addi-tubulin , which wo-semiquinone radical. This radical is extremely reactive with oxygen and autoxidizes to aminochrome under aerobic conditions. Molecular oxygen is then reduced to superoxide radicals, generating a redox cycle between the leukoaminochrome o-semiquinone radical and aminochrome [o-semiquinone radical plays an important role in aminochrome neurotoxicity because it induces an energy collapse when flavoenzymes utilize NADH, which is required for ATP synthesis in the mitochondria [Aminochrome can undergo a one-electron reduction by flavoenzymes that utilize NADH or NADPH as an electron donator to generate a leukoaminochrome-nochrome , 36. Thechondria , 38. Thechondria , 38–45 (α-synuclein protofibrils [α- and β-tubulin [Aminochrome can undergo a two-electron reduction by DT-diaphorase (EC.1.6.99.2), a flavoenzyme that uses both NADH and NADPH as electron donors, and the product of this reaction is the hydroquinone leukoaminochrome . Leukoamofibrils , 48 and -tubulin . DT diap-tubulin   to 4-peroxide , 51. Thenochrome . All gluomelanin –55. GST nyl dopa . Melanomnyl dopa . Thus, tα-synuclein is degraded by chaperone-mediated autophagy (CMA) and macroautophagy because the inhibition of CMA and macroautophagy lead to accumulation of wild type α-synuclein [α-synuclein mutant A53T induces CMA dysfunction, which is mediated by the expression of α-synuclein protofibrils [α-synuclein mutants inhibit their own degradation and that of other substrates [α-synuclein mutant A53T in transgenic animals demonstrates that A53T localizes to mitochondrial membranes as a monomer, inhibiting complex I and increasing mitochondrial autophagy [α-synuclein impairs autophagy via Rab1a inhibition, and Rab1a overexpression rescues the autophagy defect caused by α-synuclein [α-synuclein [Autophagy is an important intracellular bulk degradation and recycling process in which cytoplasmic proteins and organelles accumulate in autophagy vacuoles that are transported into lysosomes –60. Autoynuclein . The expofibrils . Interesbstrates . Overexputophagy . α-synucynuclein . A mutatynuclein . Parkin ynuclein . The losynuclein . Transfeynuclein . Autophaynuclein  and atteynuclein .α-synuclein, UCH-L1, and DJ-1 [α-synuclein (A53T) generates protofibrils that inhibit CMA autophagy. As a result, dopamine-modified α-synuclein is poorly degraded by CMA and also inhibits the degradation of other substrates using this pathway [α-synuclein by binding to the 125YEMPS129 motif of α-synuclein and inducing and stabilizing the formation of protofibrils [α-synuclein protofibrils, such as the A53T mutant (In Parkinson's disease, autophagy dysfunction plays an important role in the neurodegeneration of dopaminergic neurons containing neuromelanin. Proteins associated with familial PD have been reported to play a role in autophagy dysfunction, such as and DJ-1 , 67, 69. pathway . Aminochofibrils . These oT mutant . Mutatedα- and β-tubulin and normally exist as dimers. Microtubules also play a role in the formation of autophagosomes and fusion of autophagosomes with lysosomes [β-tubulin [α- and β-tubulin network and its aggregation around the cell membrane ([α- or β-tubulin (μM aminochrome in either the presence or absence of 100 μM dicoumarol induced a significant increase in the number of autophagosomes in the cytosol (6- and 9-fold, resp.; α- or β-tubulin, preventing the fusion of autophagosomes and lysosomes that lead to an increase in the number of autophagosomes in the cytosol [Microtubules are an important component of the cytoskeleton, which are composed by subunits of ysosomes , 75. Ami-tubulin  and the embrane (; Figure -tubulin . Incubat cytosol . Further cytosol . Aminoch cytosol  as well  cytosol . All of


Recent experiments reveal both passive subdiffusion of various nanoparticles and anomalous active transport of such particles by molecular motors in the molecularly crowded environment of living biological cells. Passive and active microrheology reveals that the origin of this anomalous dynamics is due to the viscoelasticity of the intracellular fluid. How do molecular motors perform in such a highly viscous, dissipative environment? Can we explain the observed co-existence of the anomalous transport of relatively large particles of 100 to 500 nm in size by kinesin motors with the normal transport of smaller particles by the same molecular motors? What is the efficiency of molecular motors in the anomalous transport regime? Here we answer these seemingly conflicting questions and consistently explain experimental findings in a generalization of the well-known continuous diffusion model for molecular motors with two conformational states in which viscoelastic effects are included. After the publication of Albert Einstein’s theory of Brownian motion in 1905 The intracellular fluid (cytosol) of biological cells is a superdense same motors in the same cell? Which role is played by the size of the cargo, and what determines the precise behavior of such active transport and its efficiency? These are the questions we answer in this article.What happens to the active motion of particles in the cytosol which are driven by molecular motors A well-established physical approach to anomalous transport phenomena is based on the intrinsic viscoelasticity We study the interplay of the viscoelastic environment of the cytosol with the action of a molecular motor and its cargo. A well-established model of Brownian motors of the kinesin family is based on the continuous diffusion of a Brownian particle in a potential landscape, which randomly fluctuates in time between two realizations, Within the power-stroke idealization, the maximal mean velocity of the motor due to the fluctuations between the potentials Both the motor and its cargo are subjected to friction and random thermal forces from the environment. For normal viscous Stokes friction, the frictional drag force is The above ratchet model is appropriate to describe the motor dynamics at dilute solvent conditions in vitro. Our focus here is different, as we want to study the motor action in living cells, where the following experimental facts have been established: Even in the absence of cargo the effective friction coefficient for the motor is enhanced by a factor of 100 to 1000 in the cytosol compared with the one in water Methods for details. On the basis of this theoretically and experimentally well-founded approach the results herein were obtained from numerical analysis.Methods) and The first major surprise is that even carrying large cargo particles like magnetic endosomes with radius about 300 nm our model motor can operate by an almost perfect power stroke mechanism in the normal transport regime, as demonstrated in Indeed, when we decrease the potential amplitude by Depending on the cargo size and the binding potential amplitude the transport can become more normal and thermodynamically highly efficient even for a large turnover frequency, as ven for  . Still, For vanishing loading force We proposed a simple basic model which reconciles experimental observations of both normal and anomalous transport by highly processive molecular motors in biological cells. Our model presents an immediate generalization of a well-known two-state model of normally diffusing molecular motors by accounting for the viscoelastic properties of the intracellular fluid. It not only explains how molecular motors may still operate by a power-stroke like mechanism while carrying a large cargo which subdiffuses when left alone, but also why and how an anomalous transport regime emerges for even larger cargo. It is crucial for this explanation that viscoelastic subdiffusion and anomalous transport possess finite moments of residence times in any finite spatial domain. Thus, there exist time scales for sliding down towards the potential minimum within one period of the flashing ratchet potential, for the escape to another potential well, and for the mean turnover time of the potential flashes. When the time scales are well separated, the transport is normal. However, when the sliding time scale starts to interfere with the turnover time, or the interwell potential barrier is lowered so that backsteps can occur, the transport becomes anomalous. These qualitative basic features are expected to survive in more complex models of molecular motors operating in viscoelastic environments.Specifically, we showed that transport by molecular motors becomes anomalous for large cargo particles with large fractional friction coefficient Our research provokes a number of follow-up questions. Thus, what happens if we relaxed the assumption of a rigid motor-cargo linker molecule? In that case, the large subdiffusing cargo is elastically coupled to a molecular motor, that possibly still operates normally in the absence of cargo. We are currently investigating this generalization for realistic spring constants of the linker. However, qualitatively the results remain very similar. Another question is prompted by the experimental results in Ref. Our model presents a good starting point for future research and further generalizations. Understanding how molecular motors perform in the viscoelastic cytosol of living cells despite the subdiffusion of the free cargo is compelling. Our findings open new vistas to the old problem of intracellular trafficking, reconciling seemingly conflicting results for the motor-cargo dynamics under different conditions. Finally, our results will be of crucial importance for the design of new technologies of motor-driven particles and drug delivery in the crowded cytosol of cells. We are confident that our findings will prompt a series of new experiments on the dynamics of molecular motors under realistic conditions in living cells.The numerical approach to integrate the generalized Langevin The rate constants The power exponent of anomalous diffusion was fixed to


Background. The prevalence of Kaposi's sarcoma (KS), an AIDS-defining illness, has increased in parallel with the HIV/AIDS epidemic. The presence of violaceous skin lesions should raise suspicion of KS. However, especially on dark skin, KS mimics a variety of non-KS skin conditions. Histologically, there is a wide range of expressions of KS and a large number of mimickers. For all these reasons, a HHV-8 immunohistochemically biopsy-proven diagnosis of KS should be the gold standard. Methods. Prospective study of 490 consecutive skin biopsies from the general community in the Limpopo Province of South Africa, from April 2010 through December 2011. Results. The clinical discordance rate (over-/underdiagnosis of KS) was 30.5%; the histological discordance rate was 9.2%. Conclusion. Because of the magnitude of diagnostic error, both clinical and histological, all clinical lesions suspicious of KS should be biopsied and HHV-8 LAN-1 immunophenotyped. Kaposi's sarcoma (KS) became an AIDS-defining illness at the outset of the human-immunodeficiency-virus-(HIV-) induced acquired immunodeficiency syndrome (AIDS) endemic and pandemic –3. ThereSub-Saharan Africa in general and South Africa in particular are at the epicenter of the HIV/AIDS endemic. Although the prevalence rates vary somehow according to the sources, it was estimated that in 2010 it infected 17.8 percent of the South African population and up to 19.7 percent of females aged between 15 and 45 years [The human herpes virus-8 (HHV-8) or Kaposi's sarcoma associated herpes virus (KSHV) was identified as the infectious agent of all types of KS  . The latBecause of prognostic and therapeutic implications of a correct diagnosis, it is recommended to biopsy any skin lesion susceptible of being KS, especially in HIV/AIDS endemic regions. We report an observational cohort of 490 consecutive skin biopsies in HIV-infected and noninfected patients. All biopsies were immunostained with HHV-8 LANA-1; 261 (53.3%) were biopsy-proven cases of KS among a group comprising 248 (50.6%) HIV-infected patients. We investigated the rates of clinical and histological diagnostic proficiency of KS and of its clinical and histopathological mimics.The study was carried out in the Limpopo Province of South Africa. All surgical pathology specimens of public health facilities are centralized at the National Health Laboratory Service in the provincial capital, Polokwane. It serves an essentially rural population in excess of 5 Million. The study was approved by the research ethics committee. All the cases were treated anonymously. The cases were collected from April 2010 through December 2011. All the patients were of African ethnicity. The following information was recorded: age, gender, HIV status (when available), ART status, CD4+ T lymphocyte count, site of biopsy, and clinical provisional diagnosis. Only 25 biopsies were referred by specialist dermatologists.All of the cases were routinely streptavidin-biotin-peroxidase immunostained with diaminobenzidine using a murine monoclonal antibody directed against the C-terminus of the LAN-1 molecule of HHV-8 . The initial histopathological diagnosis was made before LAN-1 immunostaining and corrected accordingly when indicated in retrospect. The clinical and histopathological diagnostic proficiency was expressed in terms of concordance or discordance. Discordance was further subdivided into overdiagnosis (non-KS) and underdiagnosis (KS missed).t test, and 95% confidence intervals (CI) of proportions. The level of significance was set at P < 0.05.Statistical analysis was performed using column statistics, Student's 3 .Out of 490 cases, 261 (53.3%) were HHV-8 proven cases of KS. The rate of documented HIV positivity among the 490 cases was 248 (50.6%). Eighty-one (32.7%) were documented to be on ART and 24 (9.7%) not to be on ART. The average CD4++ count was 259.0 ± 181.6 cells/mmAmong the 261 cases of KS, 161 (61.7%) were known to be HIV-infected; no HIV serostatus was known for the 100 others. Among the KS with unknown HIV status, 52 were males and 48 females. One KS only was HIV negative at the time of biopsy.The anatomical site of KS, illustrated in n = 9), pyogenic granuloma (n = 5), and seborrheic keratosis (n = 8) were misdiagnosed as nodular stage and early patch KS, respectively.The histological rate of discordance, shown in Kaposi's sarcoma is a multifaceted entity in terms of the population groups (ethnic and gender) it affects, in terms of clinical signs and symptoms, in terms of the body parts it affects , and in terms of histopathological expression. Traditionally, the six histological variants are the early patch stage, the nodular stage, the plaque stage, the intravascular, the lymphangioma-like, and the angiosarcoma-like type . RecentlIn sub-Saharan Africa, KS accounts for about 10.0% of HIV-related malignancies and may be the first manifestation of HIV . As suchClinically, virtually all pigmented skin lesions are part of the differential diagnosis of KS . In blacThe increasing prevalence of HIV infection in sub-Saharan Africa together with the reluctance to be tested makes it difficult to distinguish the endemic KS from the AIDS-associated variant . The disThe present study shows a relatively high discordance rate between the clinical and the biopsy-proven diagnosis of KS, indicating that “violaceous” skin lesions in an HIV endemic population warrant a biopsy. The relatively high histological discordance between the pre- and post-immunohistochemical diagnosis of KS warrants HHV-8 immunophenotyping to ascertain the diagnosis.