The rate of revision for some designs of total hip replacements due to idiopathic aseptic loosening has been reported as higher for women. However, whether this is environmental or inherently sex-related is not clear.e.g. metal vs polymer)? The objective of this study was to test for material dependent inflammatory osteolysis that may be linked to sex using CoCrMo and implant grade conventional polyethylene (UHMWPE), using an in vivo murine calvaria model.Can particle induced osteolysis be sex dependent? And if so, is this dependent on the type of implant debris  or CoCrMo particles (0.9um ECD) or received sham surgery. Bone resorption was assessed by micro-computed tomography, histology and histomorphometry on day 12 post challenge.Female mice that received CoCrMo particles showed significantly more inflammatory osteolysis and bone destruction compared to the females who received UHMWPE implant debris. Moreover, females challenged with CoCrMo particles exhibited 120% more inflammatory bone loss compared to males (p<0.01) challenged with CoCrMo implant debris (but this was not the case for UHMWPE particles).We demonstrated sex-specific differences in the amount of osteolysis resulting from CoCrMo particle challenge. This suggests osteo-immune responses to metal debris are preferentially higher in female compared to male mice, and supports the contention that there may be inherent sex related susceptibility to some types of implant debris. The 1). CoCrMo particles had a median diameter of 0.88 µm diameter ECD number-based . UHMWPE particles had a median diameter of 0.95 µm diameter ECD number-based . These CoCrMo alloy an UHMWPE particulate sizes and shapes of particulate debris produced from comercially available implants have been shown to be clinically relevant and able to induce inflammatory responses in innate immune cells [The particle sizes of CoCrMo alloy particles  were produced from a commercially available total hip arthoplasty head component , using proprietary cryomilling . Particulate size was characterized by using low angle laser light scattering (LALLS) and Scanning Electron Microscopy (SEM), Fig. . To keep the samples in position during scanning, the skulls were placed in a tightly fitting rigid plastic tube filled with 10% neutral buffered formalin. To determine accurately the region of interest (ROI), a three-dimensional rendering of the whole sample was performed using the manufacturer´s software. If osteolysis or particles were visible, the ROI was defined as a cylinder with a diameter of 3.84mm and a thickness of 1.47mm beginning from the first slide that captured the bone surface in the ROI. The particle artefact was not observed and did not affect bone fraction calculations. The specific morphometric parameters bone volume (BV) and tissue volume (TV) were measured with a threshold of 350 HU (Hounsfield unit) and the BV/TV ratio was calculated.2.3® .Four-micrometer thick sections of calvaria were collected at the depth at which the presence of particles was detected within the calvarial tissue. The sections were mounted on glass slides and subsequently HE staining was performed. The HE-stained specimens were photographed digitally using a standard high-quality light microscope . The image was oriented with the midline suture in the middle of the field. The sections were coded and blinded prior to analysis. Histomorphometric measurements were performed with the image analysis software AperioThe program was calibrated using a calibration micrometer. The region of interest to be measured was defined as the area within a distance of 2 mm around midline suture. In this area the volume of the bone stock and potential osteolysis were determined and the relative ratio of osteolysis to the bone stock volume (Osteol./BV) calculated. The mean values per animal of each available section  were calculated.2.4Data is graphically shown as the mean with standard deviation. The Kolmogorov-Smirnov-Test was used to establish normality (p≤ 0.05) for n=18 male and n=18 female mice. ANOVA was used to establish significant differences for multigroup comparison (p<0.05). Subsequent statistical comparison was conducted with Student`s t-test where p<0.05 was used to establish significance. The software SPSS 22  was used to carry out the statistical computations. Summary statistics of data are expressed as means and standard deviations.32 and 3).We observed significant differences for all histomorphometric  and micro-CT parameters (BV/TV) in all animals that received UHMWPE or CoCr particles on the calvaria in comparison to the sham control groups . Example 3D reconstruction of bone within standardized regions clearly illustrates the increase in osteolysis associated with Female vs. Male mice challenged with CoCrMo particles . Using histomorphic measures of bone loss of bone volume (BV) to total standardized cylindrical volume (TV) of  and micro-CT parameters (BV/TV) differences within groups were compared . Males demonstrated greater bone volume compared to females with sham surgery only, indicating slight but significant sex differences in calvaria bone volume for age matched controls. In comparison to these baseline sham controls, both metal (CoCrMo) and plastic (UHMWPE) particles induced inflammatory bone loss (p<0.05), where the greatest amount of bone loss was observed in female mice challenged with CoCrMo particles.Selected cylindrical control volumes of mouse calvaria encompassing the placement of particle challenge upon the exposed calvaria were analyzed for total bone volume and compared to sex matched controls for volume of bone loss. 3D reconstructions of bone volume (BV) within these standardized cylindrical regions of total volume (TV) were used for quantitative analysis of the murine calvaria bone loss . This analysis of the data revealed that females challenged with CoCrMo particles had significantly more osteolysis than did any other group with greater than 100% more bone loss than males treated with the same amount of CoCrMo particles demonstrating the severe sex effect of CoCrMo particles when compared to plastic UHMWPE particles of similar size and dose. All other challenged groups normalized to sham surgery controls  did not significantly differ. Similar results were obtained for histomorphometric  analysis of H&E stained sections , where there was over 120% increase (p<0.01) in the amount of areal calculated bone loss for female CoCrMo challenged calvaria compared to males with CoCrMo. Histomorphometric  analysis also demonstrated a similar lack of sex difference between UHMWPE treated groups, supporting a metal/sex specific immunogenic mechanism of inflammatory bone loss.On a normalized basis to sham sex matched controls , the pattern of bone loss was similar to non normalized data . An inflammatory pannus with the capability to actively resorb bone is clearly illustrated where the thickness of the inflammatory tissue was approximately 2x as thick in CoCrMo challenged calvaria when compared to the pannus induced by UHMWPE for both males and females. The particle challenge is shown isolated within the inflammatory pannus and excluded from the bone interface, yet a high degree of cell infiltrates can be observed associated with pits of active bone resorption. The composition of the cell/tissue infiltrates (pannus) co-localized with pits of bone loss generally showed vascularized soft tissue fibroblasts with an abundance of macrophage or macropahge-like cells migrating into boney areas. There was no evidence of extensive lymphocyte infiltrates or aseptic lymphocyte vasculitis associated lesions (ALVAL)histology that would be associated with an adaptive immune responses. Additionally, there was no evidence of active multinuclear osteoclasts inside the calvaria responding to inflammatory stimulus and resorbing bone from the inside out . Qualitatively there was a 2x greater foreign body response associated with female murine calvaria compared to males when challenged with either UHMWPE or CoCrMo particles .Over 15 histologic sections per treatment group were analyzed at the midline suture of the calvarial bone plates proximal to particle challenge used for histomorphometric analysis. Qualitative analysis revealed a relatively increased inflammatory pannus associated with CoCrMo 5). Instead macrophages, fibroblasts and osteoclast-like cells were observed at the sites of bone resorption and eminating from the inflammatory pannus  (Note:Bars indicate 0.1mm). Neogenic woven bone can be seen in both UHMWPE and CoCrMo challenged calvaria  indicative of the compromise time period between maximal inflammatory osteolysis and maximal neogenic bone, used in this investigation.At high magnification there was no evidence of multinucleated osteoclasts at the bone pannus interface of particle treated calvaria . Instead this evidence supports a more complex picture where toxicity  and/or adaptive immune reactivity  [Our results indicate that CoCrMo implant debris induces significantly increased osteolysis in females over that of males using a murine model of particle induced osteolysis. However, not all implant debris types induced sex based differences in osteolysis using this model. Our results indicate that UHMWPE particles do not induce significantly sex-based differences in either soft-tissue inflammation or resulting osteolysis. This is consistent with clinical observations that have reported women can suffer higher failure rates of some metal-on-metal THA designs under conditions of elevated metal release . Howeverin vitro -19, whicin vitro , as wellin vitro , 21, 22 in vitro -25, bothignaling , 26, 27.itivity) , 23-25 min vivo murine model to approximate human particle induced osteolysis affected by sex and different implant materials. The compromise time point between maximal osteolysis and maximal bone-neogenesis, was practically limited to a single point at 12 days post-op. While this time point represents a combination of osteolysis and osteogenesis in the murine model, it precludes determination of whether the observed differences were due to sex based inflammatory osteolysis or osteogenic differences or a combination of both. Additionally, a cross sectional time of 12 days may have acted to obfuscate osteolysis differences for UHMWPE that may be significantly apparent at earlier time points, i.e. 5-7 days post operatively . Thus, further investigation with earlier time points and greater number of subjects may help reveal more subtle sex differences in UHMWPE. However, the results of this study support our hypothesis to some degree, indicating that sex-based differences could be detected in response to CoCrMo particulate metal debris. This is the first report demonstrating the ability of metal CoCrMo-alloy implant debris particles to induce sex dependent osteolysis differences at time points where UHMWPE did not. Because evidence for the clinical analogue of this sex-dependence has been reported for patients with metal debris in metal-on-metal hip arthroplasty revisions [There were important limitations of using an evisions , 29, theWe found sex-based influence on inflammatory osteolysis/osteogenesis reactions to CoCrMo particles (but not UHMWPE), implying increased female osteo-immuno reactivity to metallic (CoCrMo) but not polymeric (UHMWPE) implant debris. This may explain one aspect of population dependent implant performance in the outcome of orthopedic implants that preferentially release Cobalt-alloy metal debris. Further research targeting specific mechanisms of toxicity and immune responses (innate vs adaptive) that may mediate sex dependent outcomes in joint replacement surgery is needed to support these findings.


We also found that glabridin could attenuate the inhibitory effect of taurocholate on type I interferon signaling by depleting the level of cell-surface NTCP. These results demonstrate that our screening system could be a powerful tool for discovering drugs targeting HBV entry.Sodium taurocholate cotransporting polypeptide (NTCP) is a major entry receptor of hepatitis B virus (HBV) and one of the most attractive targets for anti-HBV drugs. We developed a cell-mediated drug screening method to monitor NTCP expression on the cell surface by generating a HepG2 cell line with tetracycline-inducible expression of NTCP and a monoclonal antibody that specifically detects cell-surface NTCP. Using this system, we screened a small molecule library for compounds that protected against HBV infection by targeting NTCP. We found that glabridin, a licorice-derived isoflavane, could suppress viral infection by inducing caveolar endocytosis of cell-surface NTCP with an IC Hepatitis B virus (HBV) is the causative agent of chronic hepatitis B (CHB), which can lead to liver cirrhosis and hepatocellular carcinoma. The World Health Organization reported that over 240 million people worldwide have chronic HBV . DespiteIn the HBV life cycle, the hepatitis B surface antigen (HBsAg) initially attaches to heparan sulfate proteoglycans on the host cell surface . This atin vitro and in vivo, and is currently undergoing a phase II clinical trial for chronically HBV-infected patients -taurocholic acid (TCA) in a sodium-free or sodium-containing buffer at 37°C for 15 minutes to allow TCA uptake into cells. Cells were washed and intracellular radioactivity was measured using a liquid scintillation counter.Cells were treated with [t test with Prism 6 software (GraphPad).All graphs represent means and standard deviations. The statistical significance of differences between two groups was tested using a two-tailed unpaired


We report a case of combined central retinal vein occlusion and branch retinal artery occlusion. A previously healthy 47-year-old male presented with decreased vision in the right eye after completing a half marathon. A fundus exam and retinal imaging revealed a combined central retinal vein and branch retinal artery occlusion.In the present report, we review the literature and discuss the possible mechanisms behind combined retinal vessel occlusions.To our knowledge, this is the first reported case of combined central retinal vein occlusion and branch retinal artery occlusion following intense exercise. Combined central retinal vein occlusion (CRVO) and branch retinal artery occlusion (BRAO) rarely occur together and predominantly arise in patients over 60 years of age . FortunaA 47-year-old, previously healthy, male presented with a central scotoma in his right eye after running a half marathon. He denied any associated symptoms, including pain, photopsias, or floaters. A review of the systems was negative except for a family history of glaucoma (grandmother) and cataracts (grandmother). His best corrected visual acuity (BCVA) was 20/20 in both eyes and intraocular pressure was 24 mmHg in the right eye and 20 mmHg in the left eye. A fundus examination of the right eye revealed retinal whitening along the inferior arcade with scattered dot-blot hemorrhages near the macula and mid-periphery and dilated and tortuous veins Figure . The funThe dilated and tortuous veins, slightly delayed arteriovenous transit on fluorescein angiogram, and a few peripheral dot-blot hemorrhages of the right eye suggest that the patient had an incipient CRVO together with an inferior BRAO. The patient underwent a full hypercoagulable workup, which was negative. A carotid Doppler ultrasound was performed, and it did not show any significant narrowing or soft dislodging plaques. He was started on timolol bid in both eyes to decrease intraocular pressures.At the one-month follow-up, his vision remained 20/20 bilaterally. The retina whitening and hemorrhages had resolved. He continued to note a scotoma in his right eye, which had improved since the initial visit. He was continued on timolol bid in both eyes and was advised not to perform endurance exercises.Retinal vascular occlusions following intense exercise and dehydration are a rare occurrence. In the literature, there are few reports of isolated CRVO caused by intense exercise -3. Our rDifferent combinations of retinal artery and vein occlusion as a result of different etiologies have been reported in the literature . These iHowever, the mechanism behind combined retinal vein and branch retinal artery occlusion is less intuitive. Two plausible mechanisms have been proposed to explain such combined occlusions. The first mechanism suggests the occluded vessel (vein or artery) compressing the unoccluded vessel at the site of an arteriovenous crossing. Retinal arteries and veins share a common adventitial sheath at crossing sites, and a distended vein or artery secondary to an occlusion may result in the compression of the normal vessel at these points. The second mechanism suggests that the occlusion initially takes place in the venous system . Since tRed blood cells (RBCs) play a key role during exercise by transporting oxygen from the lungs to the tissues and delivering carbon dioxide to the lungs for expiration. Studies have shown that athletes tend to have a lower plasma viscosity and hematocrit (sports anemia) as well as less rigid RBCs as compared to normal individuals . The lowStudies have shown hemorheological alterations in patients who develop exercise-induced central retinal vein thrombosis . These pWe report the case of a young patient who developed combined CRVO and BRAO soon after completing a half marathon. We review the literature and discuss the possible mechanisms behind combined retinal vessel occlusions. To our knowledge, this is the first report of combined CRVO and BRAO following intense exercise.


Sciatic nerve injuries following total hip arthroplasty are disabling complications. Although degrees of injury are variable from neuropraxia to neurotmesis, mechanical irritation of sciatic nerve might be occurred by protruding hardware. This case shows endoscopic decompression for protruded acetabular screw irritating sciatic nerve, the techniques described herein may permit broader arthroscopic/endoscopic applications for management of complications after reconstructive hip surgery.An 80-year-old man complained of severe pain and paresthesias following acetabular component revision surgery. Physical findings included right buttock pain with radiating pain to lower extremity. Radiographs and computed tomography imaging showed that the sharp end of protruded screw invaded greater sciatic foramen anterior to posterior and distal to proximal direction at sciatic notch level. A protruding tip of the acetabular screw at the sciatic notch was decompressed by use of techniques gained from experience performing endoscopic sciatic nerve decompression. The pre-operative pain and paresthesias resolved post-operatively after recovering from anesthesia.This case report describes the first documented endoscopic resection of the tip of the acetabular screw irritating sciatic nerve after total hip arthroplasty. If endoscopic resection of an offending acetabular screw can be performed in a safe and minimally invasive manner, one can envision a future expansion of the role of hip arthroscopic surgery in several complications management after total hip arthroplasty.The online version of this article (10.1186/s12891-018-2091-x) contains supplementary material, which is available to authorized users. Sciatic nerve irritation due to an acetabular screw following total hip arthroplasty (THA) is rare; the few reported were treated with revision procedure to remove an acetabular screw . Recent An 80-year-old man complained of severe pain and paresthesias following acetabular component revision surgery via the original posterolateral approach. The patient subsequently complained of right leg pain and paresthesias. The symptoms were exaggerated during initial hip flexion and internal rotation that limited ambulation. Physical findings included right buttock pain with radiating pain to the ipsilateral lower extremity. Radiographs and computed tomography imaging Figs. , and 4 sEight months after the aforementioned surgery, the patient underwent supine hip arthroscopy without distraction on a fracture table . The operating table was tilted right side upward to increase accessibility of ipsilateral buttock. An anterolateral viewing portal and poster lateral working portal were developed without incident. For deep gluteal space visualization, a 70-degree high definition long arthroscope with adjustable length cannulas were utilized. The cannula was opened to maintain the fluid flow when utilizing the radiofrequency probe. Fluid pressure was set to 60 mmHg with intermittent pressure increases up 80 mmHg. After endoscopic greater trochanteric bursectomy, the repaired piriformis from previous revision surgery was well visualized Fig. . EndoscoAfter this endoscopic surgery, immediate hip range of motion was begun with a continuous passive motion machine. The pre-operative pain and paresthesias resolved post-operatively after recovering from anesthesia. The patient advanced to household ambulation without upper extremity aids at 1 postoperative week despite recommendations for 2 weeks of ambulation using crutches. By 3 months postoperatively, he had returned to full activities. At 6 months postoperatively, he had a negative active and passive piriformis test and he remained pleased with the outcome. Postoperative imaging included radiographs and computed tomography scans with 3-dimensional reconstruction demonstrating complete resection of the offending portion of the acetabular screw.Nerve lesions following total hip arthroplasty (THA) are disabling complications occurring in 0.06 to 2.2% of arthroplasties , 6. HardAn expandable cannula facilitated safe resection of the protruding screw tip while avoiding injury to the adjacent superior gluteal nerve and sciatic nerve. Circulating endoscopy fluid might decrease thermogenesis during metal-on-metal burring of the offending screw. In addition, generated metallic debris is removed immediately through suction system attached burr and cannula to minimize bodily retention with possible adverse consequences.The decision to remove rather than try to preserve the piriformis muscle was influenced by the surgeon’s experience with deep gluteal space exploration often performed for sciatic nerve decompression.Relevant suggestions and pearls are summarized in Table For this patient, resection of offending screw tip using endoscopy was considered first, because the acetabular component implanted revision surgery had bony ingrowth without evidence of loosening. Although sciatic nerve irritation from a protruding acetabular screw is rare and its described treatment even rarer, the endoscopic techniques described herein may have broader applications. Recent interest in hip arthroscopy along with more advanced techniques gained from the endoscopic management of sciatic nerve entrapment syndrome/deep gluteal syndrome have allowed the application of minimally invasive hip surgery for conditions once thought treatable only by open sciatic nerve exploration, screw resection and/or revision hip arthroplasty. The endoscopic exploration and screw resection described herein enables less invasive surgery permitting early joint motion, accelerated rehabilitation, and potential outpatient management. Beyond iliopsoas tenotomy, endoscopic treatment may expand to other complication of THA including protruding screws causing adjacent neurovascular compromise.A protruding acetabular screw at the sciatic notch was decompressed by use of techniques gained from experience performing endoscopic sciatic nerve decompression. Expandable cannula was used to protect superior gluteal neurovascular bundle proximally and sciatic nerve distally, followed by exposure of screw using an arthroscopic shaver.Additional file 1:Endoscopic section of acetabular screw tip to decompress sciatic nerve. Via endoscopic and intermittent multiplanar fluoroscopic visualization, further dissection was performed proximally and distally to prevent inadvertent injuries to the sciatic nerve and the superior gluteal neurovascular bundle. Partial osteoplasty of the sciatic notch with a motorized burr performed under endoscopic guidance. Endoscopic resection of the offending screw tip was performed with a 5.5 mm motorized round burr. (MP4 4460 kb)


Sperm DNA fragmentation has been associated with reduced fertilization rates, embryo quality, pregnancy rates and increased miscarriage rates. Various methods exist to test sperm DNA fragmentation such as the sperm chromatin structure assay (SCSA), the sperm chromatin dispersion (SCD) test, the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labelling (TUNEL) assay and the single cell gel electrophoresis (Comet) assay. We performed a systematic review and meta-analysis to assess the value of measuring sperm DNA fragmentation in predicting chance of ongoing pregnancy with IVF or ICSI. Out of 658 unique studies, 30 had extractable data and were thus included in the meta-analysis. Overall, the sperm DNA fragmentation tests had a reasonable to good sensitivity. A wide variety of other factors may also affect the IVF/ICSI outcome, reflected by limited to very low specificity. The constructed hierarchical summary receiver operating characteristic (HSROC) curve indicated a fair discriminatory capacity of the TUNEL assay (area under the curve (AUC) of 0.71; 95% CI 0.66 to 0.74) and Comet assay . The SCSA and the SCD test had poor predictive capacity. Importantly, for the TUNEL assay, SCD test and Comet assay, meta-regression showed no differences in predictive value between IVF and ICSI. For the SCSA meta-regression indicated the predictive values for IVF and ICSI were different. The present review suggests that current sperm DNA fragmentation tests have limited capacity to predict the chance of pregnancy in the context of MAR. Furthermore, sperm DNA fragmentation tests have little or no difference in predictive value between IVF and ICSI. At this moment, there is insufficient evidence to recommend the routine use of sperm DNA fragmentation tests in couples undergoing MAR both for the prediction of pregnancy and for the choice of treatment. Given the significant limitations of the evidence and the methodological weakness and design of the included studies, we do urge for further research on the predictive value of sperm DNA fragmentation for the chance of pregnancy after MAR, also in comparison with other predictors of pregnancy after MAR. Traditionally, the diagnosis of male subfertility is based upon the analysis of semen volume and sperm concentration, motility and morphology. Although there is a direct relationship between semen quality and pregnancy rates both in natural conception and after medically assisted reproduction (MAR), there is no definite predictive threshold for success for conventional semen parameters –4. ConveThe integrity of our genome is continuously challenged by endogenous metabolic by-products and exogenous factors. Depending on variables like cell type, cell cycle stage and the type of DNA damage, a cell has several ways to repair damaged DNA and inaccurate repair can have different consequences ,7. WhileRecent studies have highlighted the significance of sperm DNA integrity as an important factor that affects functional competence of the sperm. Therefore the detection of sperm DNA fragmentation could be clinically useful as part of fertility workup . For thiThe SCSA bases its results on (1) the DNA fragmentation index (DFI), which is the percentage in the sample that have measurable increased red fluorescence due to acridine orange attaching to a single strand portion of DNA at sites of DNA strand breaks and then collapsing into a crystal that produces a metachromatic shift to red fluorescence under exposure to blue light and (2) the percentage of high DNA stainability (HDS), which is due to excess histones and proteins other than protamines that prevent full condensation of the sperm chromatin –32.The SCD test, also known as Halo Sperm assay, estimates the level of DNA fragmentation indirectly by quantification of the amount of nuclear dispersion/halo after sperm lysis and acid denaturation to remove excess nuclear proteins ,34.The principle of TUNEL involves labelling of the 3′-ends of single- and double-strand breaks with biotinylated dUTPs. The incorporated labelled nucleotides can be quantified by flow cytometry or (fluorescence) microscopy to determine the number of (apoptotic) sperm cells containing fragmented DNA . HoweverThe Comet assay quantifies the shape of the single cell nuclei after gel electrophoresis. Small fragmented DNA has a faster rate of migration towards the anode in an electrophoretic field (tail region) as compared to larger non-fragmented DNA (head region), leading to a typical comet shape ,31.Using these tests, the percentage of sperm with fragmented DNA was shown to be comparable in idiopathic subfertile men with normal sperm parameters and in subfertile men with abnormal sperm parameters, and significantly higher in both these groups in comparison to fertile controls –37. HoweThe electronic databases Pubmed, Embase, Cochrane and CINAHL were searched from inception (September 1967) to January 2016 for articles which described sperm DNA fragmentation tests and outcome after MAR. The Medical Subject Headings terms and/or text words that were used in our search can be found in the appendix. We also manually reviewed the bibliographies of retrieved original papers and review articles. We used the preferred reporting items for systematic review and meta-analysis checklist (PRISMA) while conducting this study .Titles and abstracts of all identified studies were screened and the full paper of the preselected articles was read by two researchers (S.M. and M.C.). Both researchers extracted the data from the article independently by using standardized data extraction forms. If 2x2 tables could be constructed the study was selected for final inclusion. In the 2x2 tables, the numbers of pregnant and non-pregnant women for different sperm DNA fragmentation cut-off values were recorded. Any disagreement between the two researchers was resolved through discussion or by consultation with a third researcher (I.S.).All studies investigating the effect of sperm DNA fragmentation detected by the SCSA, the SCD test, the TUNEL assay or the Comet assay on the outcome of IVF and/or ICSI were considered eligible for inclusion. The search was restricted to studies in humans. Studies were excluded if they had no original data available for retrieval and duplicate publications were also excluded. Studies that included cycles with donor oocytes and experiments that asses sperm DNA fragmentation in specific male factor pathologies e.g. azoospermia and co-intervention experiments e.g. antioxidant treatment were excluded from analysis.The primary study outcome was ongoing pregnancy  at the 12th week of gestation). Other study outcomes were clinical pregnancy  and live birth (defined as a live-born baby ≥ 24 weeks of gestation). All outcomes were reported per cycle.Each selected study was scored for their relevance and methodological quality by using the QUADAS 2  checklist . FurtherIn order to evaluate the overall accuracy, including the whole range of possible thresholds, we used hierarchical summary receiver operating characteristic (HSROC) plots to display the results of individual studies in a ROC space, each study being plotted as a single sensitivity-specificity point. Reported estimates for sensitivity and specificity from different studies may be based on different positivity thresholds . If there are multiple thresholds reported in one study, we chose the threshold that was most comparable to the others. Based on the binomial distributions of the true positives and true negatives we calculated a summary point, with a 95% confidence interval (CI) and predictive interval by using STATA version 14 . As recommended for meta-analysis of diagnostic accuracy studies , we usedAn area under the curve (AUC) of 1 implies perfect discrimination, whereas an AUC of 0.5 means that the test does not discriminate at all . For thiIn cases where insufficient data was available to perform HSROC analyses for clinical pregnancy, (ongoing) pregnancy or live birth independently, different pregnancy outcomes were combined when minimal differences in sensitivity and specificity were found. When minimal differences in sensitivity and specificity were found, studies with different timing of the sperm DNA fragmentation test (pre- and post-wash) were combined.2 statistic, which represents the percentage of total variability across the studies that is due to heterogeneity instead of chance. Moderate heterogeneity is defined as a value < 50%  OR ("dna"[All Fields] AND "damage"[All Fields]) OR "dna damage"[All Fields]) OR ("dna"[All Fields] AND "fragmentation"[All Fields] OR "dna fragmentation"[All Fields]) AND ("humans"[MeSH Terms] OR "humans"[All Fields] OR "human"[All Fields]) AND ("comet assay"[MeSH Terms] OR ("comet"[All Fields] AND "assay"[All Fields]) OR "comet assay"[All Fields] OR "comet"[All Fields]) OR  OR SCSA[All Fields] OR ("chromatin"[MeSH Terms] OR "chromatin"[All Fields]) OR (" acridine orange" [All Fields]) OR ("in situ nick-end labeling"[MeSH Terms] OR ("situ"[All Fields] AND "nick-end"[All Fields] AND "labeling"[All Fields]) OR "in situ nick-end labeling"[All Fields] OR "tunel"[All Fields]) OR ("in situ nick end labelling"[All Fields] OR "in situ nick-end labeling"[MeSH Terms] OR ("situ"[All Fields] AND "nick-end"[All Fields] AND "labeling"[All Fields]) OR "in situ nick-end labeling"[All Fields] OR ("situ"[All Fields] AND "nick"[All Fields] AND "end"[All Fields] AND "labeling"[All Fields]) OR "in situ nick end labeling"[All Fields]) AND ("spermatozoa"[MeSH Terms] OR "spermatozoa"[All Fields] OR "sperm"[All Fields]) AND ("pregnancy"[MeSH Terms] OR "pregnancy"[All Fields]).S1 Fig(DOCX)Click here for additional data file.


Genomics has changed the way we diagnose and treat cancer . Our abiKDM3A  [However, there is more to a genome map than a linear DNA sequence and yet we might know less than we think. A recent study has published discoveries about an epigenetic factor named  JHDM2A) .The Jumonji family of histone demethylases regulates transcription by removing methyllysine modifications from histone tails. Therefore, direct DNA interaction may not be required for the biochemical demethylase function. Amore efficient way to accomplish gene target specificity is by cooperating with transcription factors that recognize promoter and enhancer motifs via their DNA-binding domains Figure . Since tA predominant signature of androgen-dependent signaling plays a key role in prostate cancer progression and its resistance to androgen-deprivation therapy, which is the primary treatment for metastatic or locally advanced disease . TranscrMany of the factors controlled by the epigenomic regulator KDM3A are well-known biomarkers such as the prostate-specific antigen (PSA). Why can we not create a simple test that tells us if we have good genes but an unfavorable epigenome? Our epigenome is highly dynamic. Epigenomic regulators, including Jumonji family members of histone demethylases, remove or add chemical marks allowing for transient gene read-outs while blocking it in the next minute . PersonaAn additional complication is that gene activity can respond to the environment. Epigenetic silencing can, for example, arise as diet-induced consequence of inflammation . Typical


The first of two studies showed FDG activity in the left deltoid and ipsilateral axillary lymph nodes explained by influenza vaccination the day prior. The second 18F FDG-PET/CT showed multiple FDG-avid lymph nodes on both sides of the diaphragm without tracer accumulation at the vaccination site. Three months later the CT was negative for lymphadenopathy within the chest or abdominal region. Although influenza vaccination is a potential source of false positive results in FDG PET studies, generalised lymph node activation post vaccination is a rare finding with only one prior published report in individuals infected with HIV-1. This case emphasizes the necessity of taking a history of vaccination prior to a FDG PET study, and consideration of a vaccine-related immune response even without evidence of tracer activity at the vaccination site when generalised FDG-avid lymphadenopathy is encountered.We report on a 59-year-old female patient with an infected vascular graft investigated with  Influenza vaccination is a potential source of false positive results in FDG PET studies -7 as 80%A 59 year old female patient presenting with two weeks of lower back pain, nausea, fatigue and chills was referred for investigation of an infected aorto-bifemoral graft. The CT component of the PET/CT study showed fat stranding and collections around the graft. The FDG component demonstrated diffuse metabolic activity surrounding the aortic graft and multiple FDG-avid foci indicating an infected graft with multiple abscesses. The FDG PET study also showed activity in the left deltoid and left axillary lymph nodes. (A) On questioning, the patient gave a history of having had the influenza vaccination the day prior.Intravenous antibiotics were started and 26 days later the patient was referred for a repeat FDG PET/CT study to assess treatment response. The PET/CT study showed reduced but persistent FDG activity around the aortic graft indicating an incomplete metabolic response to antibiotic therapy. In addition, the FDG PET/CT study demonstrated enlarged lymph nodes with FDG accumulation on both sides of the diaphragm involving the bilateral cervical and axillary, porta hepatis, coeliac and left femoral nodal stations. Interestingly, the previously documented FDG uptake in the left deltoid muscle related to the influenza vaccination had completely resolved (B). Three months after removal of the infected graft and femoral vein reconstruction, the patient was assessed by CT angiogram revealing resolution of the lymphadenopathy above or below diaphragm (C).Generalized lymph node activation following influenza vaccination is a rare and unexpected finding with only one prior published report in HIV-1-infected individuals  while abThe authors declare no conflict of interest.


Bacillus species 062011 msu is a harmful pathogenic strain responsible for causing abscessation in sheep and goat population studied by Mariappan et al. (2012) Bacillus cereus and exhibits 99% sequence homo;logy with the genomes of B. cereus ATCC 10987 and B. cereus FRI-35. Hence, we have renamed the organism as Bacillus cereus 062011msu. The Whole Genome Shotgun (WGS) project has been deposited at DDBJ/ENA/GenBank under the accession NTMF00000000 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA404036(SAMN07629099)). Specifications TableValue of the data•Bacillus cereus 062011msu is a deadly pathogenic bacterium known for causing abscess mainly in the female sheep and goat population. Hence, the genome sequence resource and their annotation details can be effectively utilized to understand the pathogenicity of the bacterium for the benefit of the farmers who rear the sheep and goat.The •Bacillus cereus 062011msu provided a broad overview regarding the subsystem features, metabolic pathways, orthologous groups, virulence factors and antibiotic resistant genes associated with the genome of the species. Most of the unique genes of the species were found to be clustered in ten genetic islands. In this study we provided a detailed analysis of the genes clustered on the genetic islands.The genome annotation data of •Bacillus species provided significant information regarding the identification and taxonomic classification of this new bacterial strain. Although according to the previous study using the partial 16S RNA sequences the pathogen was reported to be genetically similar to Bacillus anthracisBacillus cereus and phylogenetically related with B. cereus ATCC 10987 and B. cereus FRI-35.The data obtained from 16S rRNA analysis and genome sequence comparison with other •The entire genome dataset can be utilized further for determining the genes and biochemical pathways related to the pathogenicity (abscess) of the strain and developing new antimicrobial drugs for the pathogen.1Bacillus cereus 062011msu. B. cereus 062011msu. The data describing the length and Phred quality score distribution of the raw and filtered reads are illustrated in the B. cereus 062011msu genome based on RAST genome annotation. The complete list of the RAST annotated genes is given in SBacillus cereus 062011msu based on RAST annotation are listed in Bacillus cereus 062011msu resulted from BLAST genome alignment. B. cereus 062011msu obtained from DNAPlotter.The overall data represents the genome sequencing, assembly, annotation and comparative analysis of pathogenic bacteria 22.1Bacillus cereus 062011msu was isolated from the abscess tissue of the affected female sheep and goats in Maruthamputhur village near Alangulam Region, Tirunelveli District, Tamil Nadu, India https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) The 2.2Bacillus cereus 062011 msu were annotated by using the NCBI Prokaryotic Genomes Automatic Annotation Pipeline (PGAAP) http://rast.nmpdr.org/) http://www.pathogenomics.sfu.ca/islandviewer/) The draft genome contigs of Bacillus cereus 062011msu were identified by annotating the protein coding sequences against the KEGG pathway database using the BLAST2GO program Bacillus cereus 062011msu were performed by using the EggNog database  embedded within the BLAST2GO software Simultaneously the data obtained from the RAST annotation server revealed that the draft genome contains 8721 coding sequences and 472 subsystems with “Amino Acids and Derivatives” and “Carbohydrates” were the most represented subsystem features. In addition the annotated subsystem features denoted 257 genes associated with “Virulence, Disease and Defense” including 154 genes associated with antibiotics and toxicity resistance, 52 genes associated with the synthesis of antibacterial peptides, Bacteriocins, 50 genes associated with invasion and intracellular resistance and one gene associated with adhesion. The KEGG (Kyoto Encyclopedia of Genes and Genomes) biological pathways associated with the genome of Bacillus cereus 062011msu were further screened by annotating the coding sequences against the Virulence Factor Database (VFDB) B. cereus 062011msu, indicating that the flagellar proteins might play regulatory role in the pathogenicity of the bacterium. The previous in vitro experiments by Mariappan et al., 2012 reported that the pathogen was sensitive to tetracycline (TET) and ciprofloxacin (CPFX) Bacillus cereus 062011msu were screened by using the curated database, Antibiotic Resistance Genes Database (ARDB) (http://ardb.cbcb.umd.edu/) Emphasizing the pathogenic nature of the strain, the virulence factors and toxic genes residing in the genome of 2.3Bacillus cereus 062011msu based on the genome sequence comparison using RAST server were identified as Bacillus cereus AND1407 (score 544), Bacillus cereus MSX-D12 (score 406) and Bacillus cereus BAG3O-2 (score 387). Based on the local similarity of the aligned nucleotide sequences using the rapid sequence comparison tool BLAST Bacillus cereus 062011msu exhibited 99% sequence homology with the genomes of Bacillus cereus ATCC 10987, Bacillus cereus strain M3, Bacillus cereus FRI-35, Bacillus thuringiensis serovar finitimus YBT-020, Bacillus cereus strain CC-1, Bacillus cereus NC7401, Bacillus cereus AH187 respectively. In microbial genomics research the comparison of 16S rRNA gene sequence has emerged as a reliable technique to identify new bacterial strains associated with pathogenicity and infections Bacillus cereus 062011msu genome was aligned to its nearby homologs using the Clustal W multiple sequence alignment and the phylogenetic analysis was performed through the maximum likelihood method with 100 bootstrap replicates using the MEGA7 software (www.megasoftware.net/) Bacillus cereus and exhibits close evolutionary relationship with B. cereus ATCC 10987 and B. cereus FRI-35 as they were clustered together as a monophyletic clade. Simultaneously we have also performed the phylogenetic analysis based on 23S rRNA gene sequence comparison using the MEGA7 software. The 16S rRNA gene derived phylogenetic tree was found to be concordant with the 23S rRNA gene tree as it also identified B. cereus ATCC 10987 as the closest evolutionary homolog of the pathogen. The complete genomic map of Bacillus cereus 062011msu representing the GC content, GC skew graphs, coordinates and coding sequence features on both forward and reverse strands obtained from RAST annotation was generated by DNAPlotter (http://www.sanger.ac.uk/science/tools/dnaplotter) The closest neighboring strains for


We present a new and simple laser-based process to porosify thin film silicon using a pulsed laser. During deposition, we incorporate gas atoms or molecules into the Si thin film. Pulsed laser radiation of wavelength  The research on porous silicon (Si) is of current interest due to its various applications . Sensors2O2 by a less than 10 nanometers thin metal layer on the surface. Depending on the metal and the doping of Si, different porosities using different etching rates are produced due to local redox reactions [3 or CrO3) to produce porous films on top of the Si wafer. The thickness of the porous films produced by stain etching is limited to 100 nm in thickness [Porous Si is usually produced by wet chemical etching of crystalline Si wafers. The most common method and well established process is anodic etching ,6. A Si eactions . The stahickness . Bao et hickness , which CThe present contribution presents a new method using pulsed laser radiation to porosify Si thin films. Successful porosification requires incorporated gas atoms within the Si film. We show that hydrogen in plasma enhanced chemical vapor deposited (PECVD) Si and Argon (Ar) in sputtered Si fulfill this requirement and allow the porosification with pulsed laser radiation. The gas atoms serve as a source for explosively expanding gas bubbles to create porous thin film Si.a = σ is the standard deviation. Amorphous Si deposited on stainless steel substrate is fixed to an x with a constant velocity x. A laser pulse repetition frequency The rolled stainless steel (X5CrNi18-10) foil substrates of thickness As a reference, we use evaporated Si for thin film Si containing no gas atoms. Evacuation of the recipient to base pressure Winters and Kay showed the relation between sputter pressure and argon (Ar) concentration in sputtered nickel films . They mep of the sputter recipient is in the range of We deposit sputtered Si layers with a Leybold Z 550 radio-frequency (RF) sputtering system operating at a frequency + ions dissociate SiH4 molecules, resulting in thin film Si growth. Knights showed the relation of decreasing hydrogen content with increasing substrate temperature during deposition [Plasma enhanced chemical vapor deposited thin film Si contains chemically bonded hydrogen, which is usually used to saturate dangling bonds in amorphous Si to improve electrical properties of the thin film. During deposition, the plasma containing Arposition . SubstraThe Raman spectra of the thin films for different fluence Renishaw plc. Evaporation of silicon in vacuum with a base pressure e formed . The founcreases . TherefoAnnealing of the effusion . A compaent heat . ExplosiIn contrast to annealed PECVD samples, tempering of sputtered Si for The comparison of the porosity of PECVD Si with sputtered Si shows a difference in the surface structure. Only sputtered Si shows pellet formation before pore formation occurs. Apparently the type of gas and its reactivity influence the porosification process. Nevertheless, both kinds of Si contain gas atoms and show an increase in pore diameter with increasing fluence. Evaporated Si containing no gas as reference shows no pore formation after laser irradiation and both PECVD Si and sputtered Si—which contain gas atoms—show pore formation, which supports our theory for the mechanism of pore formation.We present a new method to produce porous Si thin films with pulsed laser radiation of wavelength


Aortic rupture is a rare but possible complication during spine surgery. It may manifest as severe intraoperative hemorrhage or present in a delayed manner after the formation of an aneurysm or an arteriovenous fistula. Though it is commonly encountered during anterior surgeries involving the surgical field close to the thoracic or abdominal aorta, it can also occur during a posterior surgery. Aortic injury could be associated with surgeries ranging from the commonly performed pedicle screw instrumentation to a complex three-column osteotomy. It can also occur, as in the reported case, while performing complex procedures in the presence of a pre-existing aneurysm or aortic adhesions due to coexisting infectious or inflammatory pathologies. The treatment options for such aortic ruptures range from open repair to endovascular stenting techniques. We discuss a case of an aortic rupture that occurred during a posterior vertebral column resection (PVCR) procedure performed on a 58-year-old female with spastic paraparesis secondary to tuberculous spondylodiscitis and the lessons learnt. Aortic rupture during a spine surgery is a known yet disastrous complication. It can be fatal and lead to the death of a patient on the operative table. It can occur due to pre-existing aortic aneurysms or aortic adhesions due to coexisting infectious or inflammatory pathologies, during anterior surgery while working around big vessels and rarely during posterior spinal surgery. The incidence depends on the anatomical region, surgical approach, primary pathology, implant-related issues, and the surgical expertise . Injury HistoryA 58-year-old female presented with a gradually progressive, painful kyphotic deformity of the mid back of five months duration associated with progressive spastic paraparesis, with onset two weeks before presenting to our clinic. On examination there was a tender gibbus deformity in the thoracolumbar junction with spastic paraparesis and exaggerated deep tendon reflexes. Her hematological parameters showed leukocytosis  with an elevated erythrocytic sedimentation rate (70 mm/hr). X-rays showed a focal kyphotic deformity at the thoracolumbar junction. Magnetic resonance imaging (MRI) was suggestive of spondylodiscitis at T12-L1 with epidural soft tissue components  due to massive blood transfusion. Spinal infections have been reported to be associated with spontaneous aortic rupture with or without associated aneurysmal formation . Cage miAortic rupture while performing a PVCR for spondylodiscitis has not been described before. Doing PVCR in spondylodiscitis can be a problem since long standing infection can be associated with the adherence of great vessels to the vertebral body. Our case highlights the significance of carefully evaluating the surrounding structures on the MRI preoperatively to look for adherent fragments to the aorta and leaving such fragments in situ during surgical resection to prevent surgical catastrophes.


Suspected for a while to shape the phenotype of monocytes and peripheral macrophages, HCK activity indeed regulates the evolutionarily conserved contributions of these cells to host defence, wound healing and tissue repair [According to populist dictionaries, “Hick” describes a poorly educated person of provincial existence. As much as one can draw parallels between individual characters of e repair . Recent e repair ,3.HCK ablation impaired phagocytic activity of macrophages in vitro and LPS-induced endotoxemia in vivo and reduced host-defence against Listeria parasites [de novo oncogenic HCK fusion protein in hematopoietic cells can result in acute myeloid and other leukemias, while excessive HCK activity in the “right cells at the right time” promotes macrophage and neutrophil migration, rather than causing cellular transformation [HCK gene expression or catalytic activity correlates with poorer survival for colon cancer patients.HCK is a myeloid and B-cell specific member of the SRC family of tyrosine kinases (SFKs) where the prototypical member c-SRC confers neoplastic transformation when constitutively activated. The latter not only occurred following the Rous sarcoma virus' hijacking of a carboxyl terminally truncated version of c-SRC or of viral proteins corrupting the functions of SFKs, but also as a consequence of SFKs acquiring carboxyl terminal non-sense mutations. Meanwhile, the cellular homologues of SFKs play important roles in signal transduction from growth factor and immune cell receptors, as well as of various immune-cell integrins. Accordingly, genetic arasites . In contormation . Howeverormation . AlthougMyeloid HCK activity promotes solid cancer growth by regulating polarization of macrophages and most likely neutrophils: the more HCK activity the stronger the expression of genes associated with alternative activation characterized by an anti-inflammatory M2 endotype . Indeed,While adoptive transfer experiments show that HCK affects bone marrow-derived macrophages and possibly their monocytic Ly6G precursors, it remains unclear whether HCK also regulates polarization of tissue-resident macrophages derived from a MYB-positive yolk sac precursor. However our unpublished observations suggest that HCK activity also hinders elimination of (transplanted) cancer cells by the immune system. This is likely to involve the adaptive immune system as suppression of HCK appears to augment the anti-tumor immune response conferred by immune checkpoint inhibitors on immunogenic cancer cells as well.There remains the question as to where HCK fits in to the signalling cascades that may sense a need for polarization, mediate it and ultimately execute the effect on neoplastic epithelium. HCK is physically associated with the receptors for CSF-1, the IL6-family and other cytokines implicated in regulating macrophage endotypes. Whether the physical association of HCK with leukocyte adhesion receptors may prime or reinforce macrophages for polarization by cytokines remains unknown. Strikingly, excessive HCK activity still endows an M2 endotype in the absence of STAT6 and therefore limits responsiveness to IL4/13 as the classical inducers of alternative macrophage activation [To return to the “Hick” analogy, macrophages remain highly susceptible to the poor education signals provided by adjacent cancer cells and designed to support tumor progression through enhanced nutrient supply and establishing an immune tolerant shield. As a regulator of macrophage polarization, HCK may stand up as an alternative to PI3Kγ and CSF-1R, which are pursued as clinical targets for re-educating tumorigenic M2 macrophages to adopt a tumoricidal M1 endotype. Whether medicinal chemistry can overcome the challenges of identifying small molecules inhibitors with sufficient specificity to overcome the extensive molecular similarity within the 10 members of the SFK remains to be seen.


This dataset presents the mitochondrial genome variants associated with oncocytic tumors. These data were obtained by Sanger sequencing of the whole mitochondrial genomes of oncocytic tumors and the adjacent normal tissues from 32 patients. The mtDNA variants are identified after compared with the revised Cambridge sequence, excluding those defining haplogroups of our patients. The pathogenic prediction for the novel missense variants found in this study was performed with the Mitimpact 2 program. Specifications tableValue of the data•The data identified inherited mtDNA variants associated with patients with oncocytic tumors.•The data showed some identified mtDNA variants could have functional consequences.•The data might help to detect new genetic predisposition markers for oncocytomas.1The data were presented as tables, where positions, detail changes , and the2NC_012920). The heteroplasmy were defined if a double peaks of two residues were verified at the same position in the electro-chromatograms. Pathogenic prediction were analyzed using PolyPhen2 (http://genetics.bwh.harvard.edu/pph2/) We collected 32 cases of the parafin-bedded tissues with oncocytic tumor and matched adjacent normal tissues, mtDNA were amplified by PCR using 24 previously reported pairs of mtDNA primers to cover the whole mtDNA genome as our previous work


To study if difference between the tricuspid and mitral valve opening time evaluated using cardiac magnetic resonance imaging (CMR) correlates with echocardiographic measurements of left atrial pressure (LAP).The ratio of the transmitral early inflow velocity (E wave) and the tissue Doppler mitral annular velocity (e' wave) can be used to measure left atrial pressure on the echocardiographic examination. An elevated E/e' (>15) on echo Doppler is an established measure of elevated LAP.Unlike echo, cardiac MRI (CMR) has no non-invasive measure of LAP. Normally both the atrioventricular valves open simultaneously or the tricuspid valve opens earlier than the mitral valve due to a very short right-sided isovolumic relaxation time (IVRT). Elevated LAP shortens the left sided IVRT and therefore causes the mitral valve to open earlier than expected. Also, elevated LAP indirectly leads to elevation of right heart pressures causing lengthening of the tricuspid valve isovolumic relaxation time resulting in delayed tricuspid valve opening. We investigate the role of using the time differential of mitral and tricuspid opening as novel parameter of LAP measurement on CMR.We hypothesize that elevated LAP should lead to an increased positive time differential of the mitral and tricuspid valve opening times.CMR and echo Doppler studies in 32 patients were reviewed. Echo Doppler data were reviewed to measure the mitral E wave and septal e' velocities. The LAP was calculated using the following formula:LAP = (E/e' x 1.25) + 1.912 out of 30 patients met the criteria for an elevated left atrial pressure based on the E/e' ratios and the remaining 20 patients had normal LAP.On CMR, the mitral and tricuspid valve opening times (in milliseconds) were calculated using the 2D short axis cine phase contrast images obtained at the level of the AV valves. The velocity versus time graphs and cine images were used to calculate and differences in valve opening times.The correlation between the CMR derived mitral and tricuspid valve time differential and echocardiographic E/e' derived LAP values was calculated using Spearman's rank correlation.Our study showed that there is significant correlation between the echo derived LAP using E/e' and mitral and tricuspid valve opening time differential. Spearman's rank correlation was 0.39 (p=0.028). A positive time differential  had high specificity in detecting elevated LAP (85%). However, the sensitivity was poor (50%). The positive and negative predictive values were 67% and 74%, respectively.Time difference between the openings of the atrioventricular valves is a highly specific marker of elevated left atrial pressure. This marker merits further testing to determine if it can provide quantitative information on the degree of left atrial pressure elevation.


Specifications TableValue of the data•The raw data files will allow future scientists to re-analyze the data in single- or meta-analyses.•The raw dataset will allow future scientists to extend statistical analyses.•The data fills a major gap in knowledge regarding changes that underlie bald scalp-specific manifestations in AGA.•The data is linked to transcriptional profiling and functional and pathway based analyses to clarify underlying mechanisms of disease.1The raw data files (CEL files) obtained from microarray analysis, that were used in the analysis and interpretation of our related report are available in Sample IDs are:AG5011DAG5011NAG7003DAG7003NAG7007DAG7007NAbbreviations:AG=Androgenetic alopeciaN: haired scalpD: bald scalpRNA was extracted from frozen biopsies of haired and bald scalp samples from 3 AGA-patients. Genome-wide gene expression analysis was performed using the microarray platform: Human Genome U95Av2 GeneChip  according to manufacturer׳s protocols. Biotinylated probes were fragmented and hybridized to the microarray chips . Default parameters were used to import the gene expression data to Partek Genomics Suite v6.6 . Raw data preprocessing included log2 transformation, quantile normalization, RMA background subtraction and median polish and was used for probeset summarization to scale mean expression values of all 6 arrays. The preprocessed dataset was subsequently processed to establish differentially expressed genes between bald and haired scalp using the “Gene Expression” tool within Partek Genomic Suite


The axial periodicity of rat arachnoid and dura mater collagen fibrils exposed to 910 MHz for 2 h/day for 30 consecutive days was measured by means of image analysis of electron-optical data. Such measurements were compared with those from sham-exposed animals. These measurements reveal that on exposure, the intermolecular interactions during collagen fibril assembly are affected.


Bacteroidales gene markers with human sewage and animal fecal samples  from Tha Chin and Chao Phraya watersheds. Annealing locations of PCR primers were illustrated by maps of 16s rRNA Bacteroidales genes. We also summarized previously published work on the performance of the PCR assays. For further discussion of the data presented here, please refer to Somnark et al., Performance evaluation of Bacteroidales genetic markers for human and animal microbial source tracking in tropical agricultural watersheds, Environ. Pollut. 236 (2018) 100–110.We reported modified endpoint PCR results analyzed by universal and human-, swine-, and cattle-specific  Specifications TableValue of the data–Bacteroidales-modified endpoint PCR markers could be compared with microbial source tracking (MST) studies in other geographic areas for further development of region-specific MST methods.PCR results of –Bacteroidales PCR primer maps could offer an insight into annealing regions of primers for further design of new primers or evaluating currently available primers with their performance.–A summary of PCR assays that are originally designed and adopted to other regions could serve as a database for comparing the MST method performance in different geographical areas.1Bacteroidales genetic markers from both studies that originally designed the assays and studies that adopted the designed assays to be used in another geographic location  after 50–100 mL human sewage filtration was performed with a ZR Fecal DNA MiniPrep kit . DNA concentrations were measured using a NanoDrop spectrophotometer .2.2PCR primers targeting universal and human-, swine-, and cattle-specific fecal markers were selected from both endpoint and quantitative PCR platforms . A 10-μL


Pure Sertoli cell tumours (SCTs) represent less than 1% of testicular neoplasms and malignant forms are rare. We present a unique case of a 69-year-old man who initially underwent inguinal orchidectomy for a malignant SCT. He then subsequently developed a paraumbilical cutaneous lesion which was histologically identical to the primary tumour. SCTs rarely metastasise. This is the first case of SCT with cutaneous metastasis described in the literature.


The tumor suppressor p53 is a well-characterized transcription factor that can bind gene promoters and regulate target gene transcription in response to DNA damage. Recent studies, however, have revealed that p53 binding events occur predominantly within regulatory enhancer elements. The effect of p53 binding on enhancer function has not been systematically evaluated. Here, we perform a genome-scale analysis of enhancer activity from p53-bound sequences using a series of massively parallel reporter assays (MPRAs) coupled with the assay for transposase-accessible chromatin (ATAC-Seq). We find that the majority of sequences examined display p53-dependent enhancer activity during the DNA damage response. Furthermore, we observe that p53 is bound to enhancer elements in healthy fibroblasts and poised for rapid activation in response to DNA damage. Surprisingly, our analyses revealed that most p53-bound enhancers are located within regions of inaccessible chromatin. A large subset of these enhancers become accessible following DNA damage indicating that p53 regulates their activity, in part, by modulating chromatin accessibility. The recognition and activation of enhancer elements located within inaccessible chromatin may contribute to the ability of the p53 network to function across the diverse chromatin landscapes of different tissues and cell types. The tumor suppressor p53 is a master regulator of the DNA damage response and a central line of defense against genomic instability –6. FolloDrosophila uncovered an additional intergenic p53 binding site that functions as an enhancer element  and quantified the regulatory capacity of p53-bound sequences during the DNA damage response. Moreover, we have incorporated the use of the assay for transposase-accessible chromatin (ATAC-Seq) and profiled the regulatory activity of p53 binding sites in their endogenous genomic contexts. Collectively, these data demonstrate that p53 modulates the activity of hundreds of enhancer elements throughout the genome.In addition to confirming that many p53 binding sites function as enhancer elements during the DNA damage response, our analyses uncovered several previously unappreciated aspects of genome regulation by p53. We observe that p53 is bound to enhancers in healthy fibroblasts and is poised for rapid activation in response to DNA damage. Intriguingly, we find that most p53-bound enhancers reside within regions of inaccessible chromatin. The chromatin surrounding many of these enhancers becomes accessible in response to DNA damage indicating that p53 regulates their activity, in part, by modulating chromatin accessibility. Altogether, our results provide strong evidence that the regulation of enhancer accessibility and activity by p53 is an integral component of the p53 network.Human fetal fibroblasts GM06170 (Coriell Cell Repositories) were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 15% fetal bovine serum (Life Technologies). Fibroblasts were cultured with 500 nM doxorubicin for 6 h in MPRA experiments and 12 h in ATAC-Seq experiments to induce DNA damage.Lipofectamine 3000 (Life Technologies) was used to deliver MPRA plasmid pools into fibroblasts as per the manufacturer's instructions. Fibroblasts were plated in six-well dishes at a density of 50K cells/well. Fibroblasts were transfected with MPRA plasmid pools (1 μg/well) 24 h after plating. Culture medium was replaced 24 h after transfection with fresh medium containing 500 nM doxorubicin (Sigma) to induce DNA damage. Fibroblasts were harvested 6 h post-treatment for RNA and protein isolation.Cell pellets were lysed and protein concentrations were quantified by BCA assay (Pierce). Western blots were performed on protein lysates (30 μg/well). Primary antibodies used were α-p53  and α-GAPDH (Santa Cruz). Protein was visualized with horseradish peroxidase-conjugated protein A (Life Technologies) and Supersignal developing solution (Pierce).RNA from treated fibroblasts was isolated using TRIzol (Life Technologies) as per the manufacturer's instructions. For each sample, 2 μg of RNA was reverse transcribed using SuperScript III Reverse Transcriptase (Life Technologies). RNA was treated with DNase I (Worthington) prior to reverse transcription. qPCR was performed on an ABI7900HT real-time PCR (Applied Biosystems) using FastStart Universal SYBR Green Master-Rox (Roche). Primers for TBP mRNA were supplied by Applied Biosystems. Primers for p21 were designed using Primer3. Only those primer sets that showed linear amplification over several orders of magnitude were used for quantification. Primers and PCR conditions are listed in ChIP experiments were performed as previously described . α-p53 (MPRA targeted sequencing libraries were generated directly from 50% of each cDNA reaction using PfuUltra II Fusion HS DNA polymerase (Agilent). Libraries were size-selected using Agencourt AMPure XP beads (Beckman Coulter). Prior to sequencing, the quality and concentration of each library was assessed using the Bioanalyzer (Agilent). Primers and PCR conditions are listed in MPRA targeted sequencing libraries were generated such that the first 10 bases of each read corresponded to the 10-base tag used to uniquely identify individual oligonucleotides in the MPRA library. Only those sequencing reads in which the first 10 bases matched perfectly to a tag in the MPRA library and the following 26 bases matched the expected MPRA reporter sequence were used for quantification. Expression driven by each variable sequence in the library was defined by the sum of the reads mapping to each of the 10 distinct tags corresponding to the respective variable sequence. Differential expression between untreated and doxorubicin-treated fibroblasts was assessed using DESeq2 . For anaN appeared at position i was normalized by the frequency at which base N appeared at position i. Activity contributions were rescaled such that the maximum calculated activity contribution was equal to 1.To determine the activity contribution of each nucleotide at each position within the variable region of the MPRA library, the sum of the activity (absolute fold-change in response to doxorubicin treatment) for each element in which base °C for 45 min. Transposed DNA was isolated using the MinElute PCR Purification Kit (Qiagen). ATAC-Seq libraries were generated from transposed DNA using NEBNext High-Fidelity 2X PCR Master Mix (NEB). Libraries were size-selected using Agencourt AMPure XP beads (Beckman Coulter). Prior to sequencing, the quality and concentration of each library was assessed using the Bioanalyzer (Agilent). Primers and PCR conditions are listed in Fibroblasts were treated with 500 nM doxorubicin for 12 h to induce DNA damage. Following treatment, ∼100K cells were resuspended in transposition reaction mix  and incubated at 37ATAC-Seq reads were aligned to the human hg19) genome using Bowtie2 . Aligned genome uRNA sequencing reads were mapped to UCSC known genes and our previously described catalog of noncoding RNAs using TopHat2 with default options ,21. DiffP-values were corrected for multiple hypothesis testing using the Bonferroni method. Significance analysis of transcription factor binding site co-occurrence within pioneering sites was calculated using the hypergeometric distribution given the number of pioneering sites overlapped by each factor under comparison and the total number of pioneering sites in the genome. Resulting P-values were corrected for multiple hypothesis testing using the Bonferroni method.Enrichment of transcription factor (TF) binding within pioneering sites was calculated using the formula: enrichment = (s/S)/(g/G); where s = the number of TF peaks overlapping a pioneering site, S = the number of nucleotides covered by pioneering sites/the number of nucleotides covered by TF peaks, g = the number of TF peaks in the genome, and G = the number of nucleotides in the genome/the number of nucleotides covered by TF peaks. To calculate statistical significance of enrichment the labels of all TFs were randomly shuffled and the analysis was repeated 100 times. The resulting Human transcription factor binding sites were obtained through the transcription factor ChIP-Seq Clusters Version 3 track from the UCSC Genome Browser.To systematically evaluate the capacity of p53-bound sequences to function as regulatory enhancer elements during the DNA damage response we designed and performed a series of massively parallel reporter assays (MPRAs). The MPRA is a high-throughput sequencing-based method for simultaneously characterizing the regulatory potential for thousands of defined DNA sequences in a single experiment ,23. For For the MPRA oligonucleotide pool we designed 5,700 oligonucleotides with variable regions corresponding to the genomic sequences of 570 p53 binding sites throughout the human genome Figure . In paraTo determine which of the variable regions in the MPRA oligonucleotide pool can function as enhancer elements we designed an expression vector system in which the variable regions are placed immediately upstream of a reporter gene promoter . In this two-step cloning approach the oligonucleotide pool is first cloned into an empty (no reporter gene) plasmid backbone Figure . The resTo measure the regulatory activity of variable regions within the MPRA oligonucleotide pool during the DNA damage response the Promoter–Proximal MPRA vector pool was transfected into GM06170 primary human fibroblasts. Transfected fibroblasts were then cultured in the presence of 500 nM doxorubicin for 6 hours to activate the p53 response to DNA damage. Treatment with doxorubicin resulted in a robust increase in the expression of p53 protein Figure . FurtherIn order to monitor expression levels from the Promoter-Proximal MPRA vector pool we next performed targeted RNA-Seq. We generated sequencing libraries by amplifying the region of the GFP reporter transcripts containing the unique barcode sequences. The representation of each barcode within the sequencing library provides a digital readout for the regulatory activity of the upstream genomic element. As a positive control we evaluated the regulatory activity of sequences within the MPRA pool corresponding to the p53 binding site within the p21 promoter. Treatment with doxorubicin resulted in a significant increase in expression from Promoter-Proximal MPRA vectors containing the endogenous sequence of this p53 binding site Figure . In contInterestingly, we observed that expression from Promoter-Proximal MPRA vectors containing the endogenous p21 promoter sequence was higher than their scrambled counterparts even in untreated fibroblasts Figure . This obWe next evaluated the expression from all of the sequences represented within the Promoter-Proximal MPRA vector pool. We found that the vast majority of sequences corresponding to p53 binding sites display increased regulatory activity in response to DNA damage Figure . This reTo further understand the sequence properties of p53-bound regions that drive regulatory activity we calculated the sequence activity contribution of each nucleotide in each position of the MPRA oligonucleotide pool. Our analysis identified the p53 recognition motif as the primary determinant of regulatory activity Figure . We searEnhancer elements can be separated from the genes that they regulate by varying genomic distances. To evaluate the ability of p53-bound sequences to function as enhancers when located more distant from transcription start sites we designed a Promoter–Distal MPRA vector pool. In this design the MPRA oligonucleotide pool is cloned ∼800 nt downstream of the transcription start site of a GFP reporter gene Figure . As a reThe Promoter-Distal MPRA vector pool was transfected into GM06170 primary human fibroblasts, which were subsequently cultured in the presence of 500 nM doxorubicin for 6 h to activate the p53 response to DNA damage. We then performed targeted RNA-Seq to monitor expression from the Promoter-Distal MPRA vectors. As a positive control for enhancer activity we again focused on sequences within the MPRA pool corresponding to the p53 binding site within the p21 promoter. Expression from Promoter–Distal MPRA vectors containing the endogenous sequence of this p53 binding site was significantly increased in response to doxorubicin treatment, although not as robustly as with the Promoter–Proximal MPRA Figure . PromoteWe next monitored the regulatory activity from all of the sequences represented within the Promoter–Distal MPRA vector pool. We observed that most of the sequences corresponding to p53 binding sites display increased enhancer activity in response to DNA damage Figure . This inTo further understand how the distance between a p53-bound sequence and a reporter gene promoter affects enhancer activity we compared the results from the Promoter–Proximal and Promoter–Distal MPRAs. In general, we found that the magnitude of enhancer activation in response to doxorubicin treatment was less pronounced using the Promoter–Distal MPRA approach Figure . HoweverHaving established that p53-bound sequences have the capacity to function as enhancer elements using MPRA enhancer screens, a transient reporter approach, we next characterized the regulatory potential of endogenous p53 binding sites using the assay for transposase-accessible chromatin using sequencing (ATAC-Seq). Briefly, ATAC-Seq utilizes transposon integration in live cells as a method for assaying chromatin accessibility . FollowiWe cultured GM06170 fibroblasts in the presence of 500 nM doxorubicin for 12 h to activate the p53 response to DNA damage and performed ATAC-Seq. Across three biological replicates we achieved an average sequencing depth of 42 million mapped reads per sample in untreated fibroblasts and 62 million mapped reads per sample in doxorubicin-treated fibroblasts. In addition, <10% of the mapped reads in each sample aligned to mitochondrial DNA, a common contaminant in many ATAC-Seq experiments. Our analyses identified three distinct classes of chromatin accessibility profiles surrounding p53 binding sites Figure . We founSurprisingly, we found that 62%  of p53 binding sites occur within regions of the genome that are inaccessible in both untreated and doxorubicin-treated fibroblasts Figure  and B. TTo further understand how pioneering activity at p53 binding sites relates to pioneering activity across the genome we performed a global analysis of DNA damage-induced pioneering sites. We identified 8,219 regions in the genome that displayed clear signs of pioneering activity, 275 of which are bound by p53 Figure . BecauseTo characterize potential interactions between transcription factors bound to pioneering sites we next evaluated transcription factor binding site co-occurrence between all factors that are significantly enriched within pioneering sites. For 36 of the 37 factors analyzed we observed significant co-occurrence with two or more additional factors at pioneering sites Figure . AlthougTo confirm that p53 has the capacity to function as a pioneering factor we used existing ATAC-Seq datasets to profile changes in chromatin accessibility at p53 binding sites in IMR90 fibroblasts treated with nutlin . ImportaTo characterize the relationship between chromatin accessibility and regulatory activity in response to DNA damage we integrated our MPRA enhancer screens with ATAC-Seq profiles. Of the 570 genomic regions represented in our MPRA library 34 corresponded to regions of constitutively accessible chromatin, 445 corresponded to regions of constitutively inaccessible chromatin, and 91 corresponded to pioneering sites. An example of three proximal pioneering sites bound by p53 is shown in Figure We then compared chromatin accessibility profiles with the activity of endogenous p53 binding sites by evaluating expression changes for nearby genes in response to DNA damage. Specifically, we incorporated our previously described analysis of differential gene expression in response to treatment with doxorubicin and compared expression changes for genes with transcription start sites that occur within 2 kb (upstream or downstream) of p53 binding sites  13). Co. Co13). As an alternative method for monitoring endogenous enhancer activity we used existing datasets to profile enhancer RNA (eRNA) production at p53 binding sites. Briefly, eRNAs are noncoding RNA transcripts derived from enhancer elements and their expression is positively correlated with enhancer activity . A genomTo evaluate basal enhancer activity we compared eRNA expression levels in DMSO-treated cells at regions corresponding to p53 binding sites profiled in our study . We founOur previous analyses of both Promoter-Proximal and Promoter-Distal MPRA approaches, and more specifically at the p21 promoter, revealed that p53 is bound to response elements in untreated cells . To deteFinally, we used our MPRA enhancer screen data to compare the regulatory potential of p53 binding sites based on their endogenous chromatin accessibility profiles. We found that Promoter-Proximal MPRA vectors containing sequences located at pioneering sites display significantly greater induction in response to DNA damage than those containing sequences located within regions of constitutively accessible chromatin Figure . InteresThe regulation of gene expression by p53 has been studied extensively in the context of binding events that occur within gene promoters. Several recent studies, however, have found that p53 binding sites occur predominantly within regulatory enhancer elements ,13,14. TIn our MPRA enhancer screens we found that the vast majority of the genomic sequences that are bound by p53 display some level of enhancer induction in response to DNA damage. This enhancer activity is preserved regardless of the distance between the p53-bound sequence and the transcription start site of the reporter gene being regulated. While we did observe differences in the degree of enhancer activity between the many sequences we investigated, we were unable to identify any specific sequence features that explain these variations. In fact, the only distinguishable sequence property associated with enhancer activity during the DNA damage response was the presence of a p53 recognition motif. In the absence of a p53 recognition motif responsiveness to DNA damage was completely ablated, further demonstrating that p53 is the sole factor that modulates enhancer activity. Supporting our observations, an independent report published during the course of our study described the use of multiplexed reporter assays to profile enhancer activity from p53-bound regions and identified a large class of enhancers that are characterized by single p53 binding sites .Interestingly, our enhancer screen revealed that p53 is poised on response elements prior to the induction of DNA damage. We confirmed at two independent endogenous loci (one promoter and one enhancer) that p53 is indeed bound prior to the induction of DNA damage. Previous reports have shown that p53 binds to a specific subset of gene promoters and recruits components of the transcription initiation complex in healthy cells ,30,31. OWhile our MPRA enhancer screens succeeded in uncovering novel aspects of p53 biology, there were also some clear distinctions between the enhancer activities from MPRA expression vectors and endogenous p53-bound sequences. Specifically, we observed that many sequences in our MPRA oligonucleotide pool that exhibited significant enhancer induction in response to DNA damage actually corresponded to genomic regions located within constitutively inaccessible, thus inactive, chromatin. These differences demonstrate that the MPRA is a powerful tool for evaluating the functional potential of DNA sequences, but the actual regulatory capacity of an endogenous genomic sequence is ultimately dictated by the surrounding chromatin landscape. These are important considerations to take into account during the design and interpretation of MPRA-based approaches.Perhaps the most striking discovery in our study was that p53 appears to function as a pioneering factor, binding to response elements located within inaccessible chromatin and modulating changes in chromatin accessibility in response to DNA damage. In contrast to previous reports demonstrating that p53 can bind to genomic regions with high nucleosome occupancy in response to DNA damage, we observe that p53 is stably bound within inaccessible chromatin in healthy cells ,32. WhetWe propose that p53 is poised within inaccessible chromatin in an inactive state and that p53 activation  in response to DNA damage promotes chromatin remodeling Figure . In thisFinally, we present several lines of evidence demonstrating that p53 binding sites associated with pioneering activity comprise a highly potent class of p53-regulated enhancers. We find that genes located near p53-bound pioneering sites display significantly higher levels of induction in response to DNA damage as compared to genes near constitutively accessible or constitutively inaccessible binding sites. Furthermore, we observe significant increases in eRNA expression from p53-bound pioneering sites following p53 stabilization. Lastly, our MPRA screens revealed that the sequences of p53-bound pioneering sites display significantly greater enhancer activity during the DNA damage response as compared to the sequences of constitutively accessible p53 binding sites.The potent enhancer activity from p53-bound pioneering sites could not be attributed to the presence of binding sites for any specific regulatory co-factors or other sequence properties. Rather, we hypothesize that constitutively accessible genomic regions contain binding sites for a large array of regulatory factors and that p53 must compete with all of these factors to modulate enhancer activity. Conversely, sequences located within inaccessible chromatin are less likely to contain response elements for as many regulatory factors. Therefore, the elevated enhancer activity from sequences located within pioneering sites may result from the lack of regulatory competition with other factors as opposed to the presence of any single co-factor. This regulatory competition would also explain the overall variation in the enhancer activity we observed in our MPRA enhancer screens.In conclusion, our findings have shown that p53 binding sites throughout the genome can function as DNA damage-responsive enhancer elements. Moreover, we have uncovered a previously unappreciated role for p53 in the modulation of chromatin accessibility at enhancers during the DNA damage response. The ability to bind and activate regulatory elements located within inaccessible chromatin may explain how p53 can effectively mediate cellular stress responses across a wide variety of tissues and cell types that harbor diverse chromatin landscapes.The Gene Expression Omnibus accession number for the MPRA and ATAC-Seq data reported in this paper is GSE83780.Supplementary DataClick here for additional data file.


A 38-year-old woman with a history of recurrent deep venous thromboses (DVTs) on chronic anticoagulation presented with acute left leg swelling. The patient was diagnosed with an acute left lower extremity (LLE) DVT in the setting of May-Thurner syndrome for which treatment with unfractionated heparin was started. Her hospital course was complicated by a new diagnosis of heparin-induced thrombocytopenia (HIT), with an incidental discovery of a large tricuspid valve mobile mass on a transthoracic echocardiogram (TTE). Subsequent imaging confirmed multiple right atrial thrombi along with LLE venous stent thrombosis and a new right LE acute DVT. Anticoagulation with argatroban for HIT thrombosis was started. She underwent a right atrial percutaneous thrombectomy and bilateral lower extremity thrombectomy with directed angioplasty and stent placement. This presentation is a rare manifestation of HIT with extensive intracardiac and deep venous thrombi, with successful staged interventions. HIT is an adverse immune reaction to heparin characterized by systemic platelet consumption and paradoxical thrombotic complications. HIT is mediated by immunoglobulins directed against platelet factor 4 (PF4) bound to heparin, which also activate platelets leading to thrombosis . The incA 38-year-old woman with a history of recurrent LLE DVTs in the setting of pregnancy and travel presented with new left leg swelling. She had a negative hypercoagulable workup and was previously treated with rivaroxaban. She had previous exposure to unfractionated heparin with no complications. Heparin infusion was started for her new LLE DVT. CT imaging revealed effacement of the left common iliac vein by the right common iliac artery suggestive of May-Thurner syndrome. Subsequent venogram revealed complete occlusion of the left external iliac vein and left common femoral vein for which thrombolysis with tissue plasminogen activator (tPA) was administered, followed by angiography with stent placement.μL to 133K/μL). Platelets continued to decline on heparin to a nadir of 52K/μL on hospital day 9  against PF4/heparin complexes is the pathophysiologic mechanism of greatest clinical concern . PlateleμL when apixaban was started for lifelong anticoagulation given her risk for recurrent DVT. She received a loading dose of 10 mg twice daily for 7 days, transitioned as an outpatient to a maintenance dose of 5 mg twice daily. She remained without evidence of recurrent thrombosis at her 2-week and 3-month follow-ups.The use of novel oral anticoagulants (NOAC) in the management of HIT is an emerging subject of study. Clinical experience, though as yet limited, has shown good platelet count responses and tolerability in patients with HIT transitioned to NOAC, both directly after heparin discontinuation and after argatroban therapy , 9. Our This patient was incidentally found to have an anatomic predisposition to venous thrombi when CT revealed evidence of left common iliac vein compression against her fifth vertebral body by her right common iliac artery. This finding is suggestive of May-Thurner syndrome, a pathologic condition characterized by LLE venous insufficiency secondary to left iliac vein compression. As the compression is usually physiologic, May-Thurner syndrome is rarely considered during the workup of recurrent DVTs; indeed the syndrome presents as DVT in only 2-3% of cases . HoweverStudies suggest that endovascular correction of pathologic insufficiency via venous stenting may improve symptoms , 13. PreHIT is an important consideration in patients hospitalized for thrombotic events with the delayed onset of thrombocytopenia while on heparin therapy. Despite the onerous potential consequences of HIT in this patient with extensive thrombosis in the setting of an anatomical predisposition to DVT, her relatively young age and excellent baseline functional capacity facilitated successful staged endovascular management of both her intracardiac thrombi and her acute DVTs.


In this video the Wright et al.  demonstrOn the flipside evidence from studying changes in the operated kidney using MAG3 renal scan also supports the notion that not every minute of ischemia counts and renal parenchyma preservation plays a more important role in renal function preservation if the warm ischemia time can be limited to less than 25-30 minutes . FurtherHomayoun Zargar, MDDepartment of UrologyRoyal Melbourne Hospital300 Grattan St, ParkvilleVIC 3050, Australiahomi.zargar@gmail.comE-mail: