This case report highlights an unusual case of sudden sensorineural hearing loss related to superficial siderosis (SS). Our patient had a craniotomy for medulloblastoma 23 years earlier, and this may represent a delayed complication related to this procedure. Magnetic resonance imaging (MRI) remains the key diagnostic investigation to illustrate the imaging features of superficial siderosis and exclude other pathologies. Increased awareness of progressive and sudden hearing complications caused by SS is important in the otolaryngologic community to expedite management and better counsel patients during the consent process. Superficial siderosis (SS) classically affects the leptomeninges, brain, brainstem, cerebellum, cranial nerves and spinal cord secondary to hemorrhage and hemosiderin deposition . Hemosid It is an uncommon condition with only approximately 100 cases reported . Early c This case report provides an account of a young male who presented with sudden sensorineural hearing loss. A 30-year-old Caucasian male patient presented to our clinic with sudden-onset left-sided sensorineural hearing loss and nonpulsatile tinnitus. His medical background was remarkable for a medulloblastoma treated with a suboccipital craniotomy and tumour resection with adjuvant radiotherapy at the age of seven. The patient had no postoperative complications or evidence of tumour recurrence on long-term followup. Examination revealed bilateral sensorineural hearing loss on tuning fork testing with normal otoscopic findings. Dysdiadochokinesia, past pointing, and intention tremor were also noted affecting the patient's left upper limb. Audiometry showed bilateral down-sloping moderate-severe sensorineural hearing loss, which was slightly worse on the left side. No prior audiometry was available for comparison.  The pathogenesis of SS results from chronic hemorrhage into the subpial layers with a predilection for the superior vermis, crests of the cerebellar folia, basal frontal lobe, temporal cortex, brainstem, spinal cord, nerve roots, and cranial nerves one and eight. The sources of hemorrhage are cerebrospinal fluid (CSF) cavity lesions, tumors, and vascular abnormalities .  The exhaustion of ferritin biosynthesis by the microglial cells as a result of iron overload subsequently causes neuronal injury . Cerebel The patient's history of previous intradural cranial surgery for medulloblastoma is a risk factor for later development of SS  and may  T2-weighted fast imaging employing steady-state acquisition (FIESTA) magnetic resonance imaging (MRI) revealed hypointense thickened bands along the vestibulocochlear nerves bilaterally which represent classic diagnostic features of SS . FIESTA  SS is a well-described delayed complication following craniotomy. Patients and their carers must be counseled regarding its association with sensorineural hearing loss, both sudden and progressive, during the consent process. Increased awareness of SS amongst the otolaryngologic community is important so that appropriate audiometric testing, imaging, and management may be expedited.


Weight suppression has been found to negatively predict treatment completion of cognitive behavioural therapy (CBT). However, subsequent attempts to replicate these findings have failed. In light of this, the current study revisits the relationship between weight suppression and treatment outcomes in bulimia nervosa. We propose that moderator effects may assist in interpreting previous inconsistency. Moderators tested were parent history of overweight, chubby/overweight childhood body shape, higher pre-treatment body mass index, and highest ever adult weight discrepancy. Participants were 117 female outpatients aged 16-54 years with bulimic disorders treated with enhanced cognitive behavioural therapy at a specialist community clinic. Logistic regression indicated that pre-treatment weight suppression did not predict drop-out or poor treatment outcome. No moderator effects were observed when hypothesised moderator features were included in treatment completion or treatment outcome models. The current study calls into question the association between weight suppression and treatment outcome. Future research into moderator models, using a larger sample, could assist us to refine conceptualizations of why some patients who have a weight suppression history are vulnerable to poor treatment adherence and outcome and to establish clinical interventions that enhance prognosis.Adult Treatment and Services stream of the 2013 ANZAED Conference.This abstract was presented in the


Lobular capillary hemangioma (LCH) is a benign lesion of vascular origin. It rarely involves nasal cavity which most commonly manifests as progressive nasal obstruction and epistaxis. In this report we present a case of LCH of the nasal cavity which occurred approximately one month after delivery. There was no recurrence after complete endoscopic resection during one year follow up.  Lobular capillary hemangioma (LCH) is a benign lesion of vascular origin. The most common sites of lesion are the skin and oral mucosa. LCH rarely involves nasal cavity which most commonly manifests as progressive nasal obstruction and epistaxis. Trauma and hormonal changes are presumed as two major predisposing factors -4. In thA 32-year-old woman came to hospital clinic with complaint of progressive right-sided nasal obstruction associated with intermittent mild epistaxis, muco-purulant rhinorrhea and post nasal drip since 10 months ago began one month after her uncomplicated delivery of a normal baby. She was completely asymptomatic during her pregnancy.. She didn’t mention any history of nasal trauma or nasal intubation but she has had history of multiple nasal packing during recent months. Endoscopic examination of nasal cavity revealed a red large polypoid mass which bled easily and originated from posterior part of middle turbinate.A CT scan of the paranasal sinuses also revealed a right nasal cavity soft tissue mass without any evidence of bony destruction or extension of the mass into the adjacent paranasal sinuses .The patient was taken to the operating room where the soft tissue mass was completely excised using endoscopic techniques. The surgery involved partial resection of the middle turbinate .Histologic finding showed proliferation of small capillaries lined by plump endothelial cells in a loboular pattern .There was no recurrence during the one-year follow-up period.Hemangiomas are vascular neoplasms that morphologically classified in to capillary, cavernous, arteriovenouse and epitheliod type . LCH, whThe etiology of LCH remains unknown but trauma and hormonal changes, such as those that occur in pregnancy, are thought to be major etiologic factors. Viral oncogenes, microscopic arteriovenous malformations, cytogenetic abnormalities and production of angiogenic factors are also associated with LCH . In fourA giant pyogenic granuloma of the posterior part of the nasal septum that occurred after prolonged use (30 days) of a nasogastric feeding tube has also been recorded . In a st Clinical symptoms include nasal obstruction, Epistaxis, Epiphora, purulant or mucous rhinorrhea and nasal deformity . In a stThe treatment of choice for these lesions is complete excision using endoscopic approach even for large lesions. In our case, there was no recurrence after complete endoscopic resection during one year follow up.


Grass pollen allergens are a major cause of allergic respiratory disease but traditionally prescribing practice for grass pollen allergen-specific immunotherapy has favoured pollen extracts of temperate grasses. Here we aim to compare allergy to subtropical and temperate grass pollens in patients with allergic rhinitis from a subtropical region of Australia.Paspalum notatum), Johnson grass  and Bermuda grass (Cynodon dactylon) as well as the temperate Ryegrass (Lolium perenne) were measured by skin prick in 233 subjects from Brisbane. Grass pollen-specific IgE reactivity was tested by ELISA and cross-inhibition ELISA.Sensitization to pollen extracts of the subtropical Bahia grass (Patients with grass pollen allergy from a subtropical region showed higher skin prick diameters with subtropical Bahia grass and Bermuda grass pollens than with Johnson grass and Ryegrass pollens. IgE reactivity was higher with pollen of Bahia grass than Bermuda grass, Johnson grass and Ryegrass. Patients showed asymmetric cross-inhibition of IgE reactivity with subtropical grass pollens that was not blocked by temperate grass pollen allergens indicating the presence of species-specific IgE binding sites of subtropical grass pollen allergens that are not represented in temperate grass pollens.Subtropical grass pollens are more important allergen sources than temperate grass pollens for patients from a subtropical region. Targeting allergen-specific immunotherapy to subtropical grass pollen allergens in patients with allergic rhinitis in subtropical regions could improve treatment efficacy thereby reducing the burden of allergic rhinitis and asthma. Grass pollens are amongst the most frequently recognised aeroallergens worldwide ,2. AllerLolium perenne) which is the major clinically relevant allergenic grass pollen in temperate regions of Australia [Paspalum notatum), Johnson grass  and Bermuda grass (Cynodon dactylon) were among the most abundantly observed grasses [Despite the clinical importance of grass pollen allergy in allergic respiratory diseases, it has not been established which of the grass pollens are most relevant to Australian environments, with the exception of Ryegrass  grass pollen extracts, house dust mites , cat hair , moulds , and Ragweed pollen .Skin prick testing were performed according to the guidelines of the Australian Society for Clinical Immunology and Allergy . BrieflyArachis hypogaea extract) was prepared as a control allergen [Proteins from non-defatted pollen grains of Ryegrass, Bahia, Timothy, Johnson and Bermuda grasses  were extracted in phosphate buffered saline with complete protease inhibitor cocktail  for three hours on a rotating wheel at 4°C and clarified as previously described . Raw peaallergen .Plasma samples diluted 1/10 were tested by ELISA with microtiter plate wells coated with 5 μg/ml of whole grass pollen extract as described previously . InhibitData was assessed for normality by Kolmogorov-Smirnov test and assessed for differences by Friedman's ANOVA with Dunn's multiple comparison tests. P values less than 0.05 (*), < 0.01 (**), or < 0.001 (***) were considered significant. Correlations of SPT and IgE reactivity between different grass pollens were determined by Spearman's rank test for paired data.Participants were recruited from amongst patients presenting to an Immunology or Respiratory clinic at a major public hospital in Brisbane, as well as amongst healthcare and laboratory workers. Seventy seven participants were skin prick test (SPT) negative to all aeroallergens tested. Eighty subjects were SPT positive to allergens other than grass pollens with house dust mite being the most frequently recognized allergen. Seventy six of the 233 participants assessed by skin prick tests showed positive reactions to at least one grass pollen extract . There were 48 grass pollen-allergic subjects for whom SPT data was available for each of Ryegrass, Bahia, Johnson and Bermuda grass pollens. Of these 48 subjects there were eight (16%) who only showed positive SPT to subtropical grasses, whereas three (6%) showed a positive SPT to Ryegrass pollen only.SPT wheals were significantly larger with Bahia and Bermuda grass pollens than with Ryegrass pollen Figure . InteresThere were significant correlations in allergic sensitivity between each of the grass pollens Table . The strIgE reactivity with each of the grass pollens was tested by ELISA in 175 subjects for whom plasma was available. The IgE reactivity of the 70 SPT-negative, non-atopic donors was used to define the normal range. Concentrations greater than three standard deviations above the mean IgE reactivity of normal subjects were considered positive; these cut off values were as follows - Bahia grass , Bermuda grass (0.129 OD) and Ryegrass (0.187 OD). Using this definition, of 50 subjects with allergies other than grass pollen, there were one, two, two and four subjects who tested positive for IgE reactivity with Bermuda, Johnson, Bahia and Ryegrass pollen, respectively. Forty five of 55 (81%) of grass pollen-allergic subjects with positive SPT to any grass pollen showed IgE reactivity with at least one of the four grass pollens by ELISA. There were ten subjects (18%) who only showed IgE reactivity with subtropical grass pollens whereas only four (7%) subjects showed IgE reactivity with Ryegrass pollen only. Mostly, subjects showed IgE reactivity with more than one grass pollen but there were differences in the levels of IgE reactivity.The IgE reactivity of the grass pollen-allergic donors was higher with Bahia grass pollens than with Johnson and Ryegrass pollens Figure . IgE reaPlasma IgE reactivity with each of the grass pollens were well correlated amongst the grass pollens Table . HoweverReciprocal cross-inhibition assays were performed in a subset of subjects known to have grown up in the Brisbane area and who would represent patients primarily exposed to subtropical grass pollens, in order to compare the strength of IgE reactivity with subtropical versus temperate grass pollen allergens Figure . The relHitherto grass pollen allergy has been primarily studied in subjects living in regions with temperate climates such as parts of Europe and North America where Timothy grass pollen is the predominant clinically important grass pollen. Ryegrass pollen has been considered to be the most clinically important grass pollen allergen in Australia ,13. HoweHere we show marked differences in the relative avidity of IgE binding between different grass pollens in dose responsive four-way reciprocal cross-inhibition experiments. Other studies of patients from temperate regions focused on inhibition of IgE reactivity with Timothy grass pollen extract or its allergenic components with an excess of pollen inhibitor and reported maximal inhibition capacity but not relative avidity as revealed by decreasing doses of pollen inhibitor ,27. PoolThere were differences in the degree of correlation of SPT sensitivity Table  and plasThere are known to be differences in allergen composition amongst grass pollens. Pollens of different species of temperate grasses all contain several major allergen families including group 1, 2, 4 and 5 whereas the subtropical grass pollens lack the group 2 and 5 allergen families . There aThe difference in allergic sensitivity and allergen recognition depends not only on biological differences between the pollens but also upon the origin of the patients investigated. Our observations with patients from a subtropical region is in contrast to IgE reactivity of patients from the temperate region city of Melbourne, who showed substantially higher IgE reactivity with pollens of Ryegrass compared with Bahia and Bermuda grasses . SimilarWe also sought to compare the allergic sensitivity and IgE reactivity amongst subtropical grass pollens. The same heterogeneity evident within temperate grass pollen allergens discussed earlier might be expected for pollen allergens of the subfamilies of subtropical grasses. Indeed we observed Johnson grass pollen elicited lower SPT and plasma IgE reactivity than Bahia grass pollen as well as lower capacity to inhibit serum IgE reactivity with other grass pollens, including Bahia grass pollen suggesting that there may be allergenic differences between these Panicoideae grass pollens. It is worth considering that although both these grasses belong to the one subfamily they align to different subtribes, Paniceae for Bahia grass and Andropogoneae for Johnson grass, which may explain why these grass pollens were not interchangeable. A difference between Bahia and Bermuda grass pollens was not evident by SPT but Bahia grass pollen showed higher levels of IgE reactivity than Bermuda grass pollen. We noted that Bermuda grass pollen appeared to be more allergenically distinct from Ryegrass than Bahia grass pollen since in three of five donors IgE reactivity with Bermuda was only substantially inhibited by itself and it had only limited capacity to block IgE reactivity with Ryegrass, Johnson or Bahia in some subjects. Also, SPT and plasma IgE reactivity with Bermuda showed the least correlation with Ryegrass suggesting greater allergenic differences between Bermuda and Ryegrass pollens than between Bahia and Ryegrass. However, further studies are warranted to confirm this observation with a wider representation of patients from subtropical regions.The outcomes of this study are likely to impact upon the clinical efficacy of grass pollen allergen-specific immunotherapy in patients from a subtropical region. Whilst for many treatment for allergic rhinitis can be managed by allergen avoidance and pharmacotherapy, for others with more severe disease, specific immunotherapy is indicated . Grass pHere we demonstrate higher SPT and IgE reactivity with subtropical grass pollens than Ryegrass in patients from a subtropical region. We also observed incomplete and asymmetric cross-inhibition of serum IgE between subtropical grass pollens and the Ryegrass pollen allergen in patients with grass pollen allergy in Brisbane. Further studies are warranted to assess difference in allergic sensitization in other parts of Australia, including arid and wet tropical regions where there are hotter climates and subtropical grass pollens are likely to contribute to grass pollen sensitization. It will be necessary in the longer term to investigate whether differences observed here in IgE recognition of subtropical and temperate grass pollen affect the efficacy of immunotherapy for patients primarily sensitized to subtropical grass pollen allergens in Australia and other regions with similar climatesELISA: Enzyme-linked immunosorbant assay; IC50: Inhibitor concentration giving 50% inhibition; IgE: Immunoglobulin E; OD: Optical density units; SPT: Skin prick test.JMD has received grants from the Asthma Foundation of Australia, Australian Society for Clinical Immunology and Allergy, The University of Queensland and the National Health and Medical Research Council of Australia, has collaborations with Sullivan Nicolaides Pathology (Queensland) and Phadia Pty Ltd , and is an inventor on an Australian and US patent on "Novel immunogenic molecules and uses thereof (Bahia grass)". JMD has also received consulting fees from Stallergenes Pty Ltd  and has a collaborative research agreement with this company. MT has received Honoraria from Novartis for asthma nurse education presentations. JWU has been paid Honoraria from Astra Zeneca to present updates on asthma at approved medical education events.JD was the senior author of this manuscript. JD designed the study, analysed data, prepared and edited the manuscript. HZL performed the ELISA experiments. MG and MT assisted with patient recruitment and skin prick testing. JWU was involved with subject recruitment and provided intellectual input to the study. All authors read and approved the final manuscript.


Monilethrix is an autosomal dominant hair shaft disorder characterized by intermittent constrictions result in short and fragility hair. We present here two afghan siblings girl, 5 and 3 years, born of consanguineous marriage, come to our department of dermatology with complains of hair loss and inability to growth long hair of the scalp since birth. When the hairs reached a certain length, they were broking. There was no relevant familial history. On examination, we found diffuse thinning of scalp hair with broken, short and spars hairs. The length of hairs was less than 2 centimeter. Multiple keratotic papules were present on entire the scalp. Hairs in eyebrows also were involved. Diffuse hair loss seen in trunk and extremity. Nails and dental were normal appearance. Other systemic examination of hair showed beaded appearance with certain interval. Histopathologic examination of scalp showed misdirection with abnormal and tortuous appearance and bulb shaped shaft in some foci considering for monilethrix. Its differential diagnosis include monilethrix-like congenital hypothericosis, alopecia areata with variable activity in the course of disease, primary cicatricial alopecia (border of fibrotic area), chemotherapy induced alopecia, monilethrix-like effect from hair styling gel and pseudo-monilethrix. There is no specific treatment of monilethrix. Improvement by hormonal treatment reported in one study that hair growths increased after first menstrual period, so it suggested hormonal influence may improve hair growths. In literatures many different options for treatment of monilethrix proposed such as oral retinoid Griseofulvin and recently topical minoxidil 2%. One study showed improvement molinethrix after treatment by iron supplementation in iron deficiency anemia.


Eliciting effective antibody responses are key to the design of HIV vaccines. While the HIV envelope protein is highly immunogenic and provokes a high-titer antibody response during viral infection and experimental immunization, affinity-matured antibodies capable of neutralizing diverse HIV isolates are rarely elicited. While recent vaccine regimens have focused on DNA, viral vector, VLP, or attenuated virus vaccines, of increasing interest are improved recombinant envelope protein immunogens. Immunizations with trimeric gp140 proteins induce higher-titer neutralizing antibody responses and have structural benefits over monomeric gp120 immunizations. Novel approaches being taken for recombinant trimeric gp140 immunogens include the use of consensus and multi-clade gp140 trimers. A consideration for these synthetic proteins that is key to vaccine efficacy: do these proteins structurally represent a native trimeric envelope?To address potential differences in the functional conformation of gp140 trimers, we evaluated the conformational characteristics of trimeric gp140 proteins from varying HIV-1 strains as well as engineered consensus gp140s via binding studies using surface plasmon resonance with a panel of well-characterized monoclonal antibodies (MAbs).Consensus trimers were recognized by more monoclonal antibodies than the primary strain gp140s, suggestive of the benefits of engineered trimers. However, analysis revealed a low trimer concentration that is competent to bind the quaternary-recognizing MAb PG9. Considering the binding to other conformational MAbs, this suggests that while the consensus gp140s contain appropriately conformational monomers and are trimeric, the vast majority of the protein is not in a proper quaternary structure.These data suggest that it is important to fully assess structural differences of immunogens even though obvious phenotypic differences may not be present. Taken together, these observations demonstrate the need to evaluate immunogens in a manner that allows the measurement of functional epitope exposure and solution conformation to assess the potential to elicit a potent, broadly-neutralizing antibody response.


FUS is a DNA/RNA binding protein found to be mutated in some cases of both sporadic and familial forms of ALS. It is still not clear how ALS-causing mutations in FUS leads to motor neuron degeneration. Here, we exploited a Drosophila model and mammalian neuronal cell lines to elucidate the role of the RNA-binding ability of FUS in regulating FUS-mediated toxicity. To determine the role of the RNA binding ability of FUS in ALS, we mutated FUS RNA binding sites  to leucines and generated RNA binding-incompetent mutants (4F-L) with and without ALS causing mutations R518K or R521C. We found that mutating 4F to L residues makes FUS RNA binding-incompetent. We observed that ectopic expression of RNA binding-incompetent FUS in fly brain, eyes and motor neurons strongly blocks neurodegenerative phenotypes as compared to RNA-binding-competent FUS carrying ALS causing mutations. Interestingly, RNA-binding deficient FUS strongly localized to the nucleus of Drosophila motor neurons and mammalian neuronal cells whereas FUS carrying ALS linked mutations was distributed to the nucleus and cytoplasm.in vivo unbiased genetic screen that led to the discovery of several dominant modifiers of mutant FUS toxicity including proteins involved in regulating SG dynamics, mitochondrial functions and RNA splicing. These genetic modifiers would help in identifying potential therapeutic targets for ALS.Importantly, we found that incorporation of mutant FUS into stress granules is dependent on the RNA-binding ability of FUS. SGs are dynamic aggregates composed of proteins and RNA that are formed when cells are under a variety of stresses. We observed that normally cytoplasmic SGs rapidly disassemble when stress conditions end, whereas cytoplasmic SGs formed in ALS patient cells having a FUS mutation fail to disassemble. This suggests that mutant FUS sequesters proteins and RNAs important for cellular homeostasis and the defect in disassembly of cytoplasmic SGs contributes to ALS. Finally, following up these observations we performed an


Pompe disease (glycogen storage disease type II) is an autosomal recessive lysosomal storage disorder caused by a deficiency of the enzyme acid alpha-glucosidase. Enzyme replacement therapy (ERT) with alglucosidase alfa has resulted in a clinical benefit in a subset of patients. Cross-reactive Immunological Material (CRIM)-negative status is associated with poor prognosis. Patients with CRIM-negative infantile Pompe disease mount a strong immune response against alglucosidase alfa ERT, resulting in a clinical decline and death despite therapy. Most develop high sustained antibody titers. Based on our clinical and laboratory experience, about 40 percent of patients with infantile Pompe disease develop HSAT. Early identification of patients at risk is needed to allow for treatment intervention with immune tolerance induction (ITI) protocols. These protocols have included the use of agents such as rituximab, methotrexate, IVIG, and agents that target the plasma cells. Different treatment approaches are needed for patients with Pompe disease treated with ITI in the naïve setting as compared to patients with high sustained antibody titers. Without ITI, the CRIM-negative patients do poorly on ERT alone. We will present a successful global experience in 15 cases and discuss the safety, efficacy and feasibility of a clinical algorithm developed at our institution to identify CRIM-negative status and allow timely initiation of ERT and ITI in this vulnerable population. Our data show that the clinical algorithm of rapidly diagnosing and initiating ITI coincident with the start of ERT in CRIM-negative patients is feasible and can be achieved in a timely manner.


Xenograft mouse models represent helpful tools for preclinical studies on human tumors. For modeling the complexity of the human disease, primary tumor cells are by far superior to established cell lines. As qualified exemplary model, patients’ acute lymphoblastic leukemia cells reliably engraft in mice inducing orthotopic disseminated leukemia closely resembling the disease in men. Unfortunately, disease monitoring of acute lymphoblastic leukemia in mice is hampered by lack of a suitable readout parameter.Patients’ acute lymphoblastic leukemia cells were lentivirally transduced to express the membrane-bound form of Gaussia luciferase. In vivo imaging was established in individual patients’ leukemias and extensively validated.Bioluminescence in vivo imaging enabled reliable and continuous follow-up of individual mice. Light emission strictly correlated to post mortem quantification of leukemic burden and revealed a logarithmic, time and cell number dependent growth pattern. Imaging conveniently quantified frequencies of leukemia initiating cells in limiting dilution transplantation assays. Upon detecting a single leukemia cell within more than 10,000 bone marrow cells, imaging enabled monitoring minimal residual disease, time to tumor re-growth and relapse. Imaging quantified therapy effects precisely and with low variances, discriminating treatment failure from partial and complete responses.For the first time, we characterized in detail how in vivo imaging reforms preclinical studies on patient-derived tumors upon increasing monitoring resolution. In the future, in vivo imaging will enable performing precise preclinical studies on a broad range of highly demanding clinical challenges, such as treatment failure, resistance in leukemia initiating cells, minimal residual disease and relapse. Preclinical mouse models are helpful tools for studying biology and therapy of diseases. Novel therapeutic approaches undergo detailed preclinical evaluation before translation into clinical trials In cancer research, a variety of different mouse models exist including xenotransplantation models and syngeneic models At best, xenotransplanted tumor cells generate a disease in mice which highly resembles the disease in men Nevertheless, sensitive follow up of leukemia progression in mice remains a limitation of the model. Invasive bone marrow aspirations in mice require prolonged periods of recuperation; blood sampling is hampered by late and heterogeneous presence of tumor cells into the peripheral blood In vivo imaging based on molecular cell marking represents a sensitive readout parameter to monitor xenotransplanted tumors in mice, e.g., using bioluminescence Here, we established the molecular labeling of patient-derived ALL cells and characterized in detail, how bioluminescence in vivo imaging enables a novel level of precision for future preclinical studies. In vivo imaging enabled quantification of treatment effects and monitoring of minimal residual disease in mice. The improved mouse model will allow performing decisive and complex preclinical studies on individual leukemias in the future.Ethikkommission@med.uni-muenchen.de, April 15/2008, number 068-08) and with the Helsinki Declaration of 1975, as revised in 2000.Written informed consent was obtained from all patients and from parents/carers in the cases where patients were minors. The study was performed in accordance with the ethical standards of the responsible committee on human experimentation .All animal trials were performed in accordance with the current ethical standards of the official committee on animal experimentation , 1 mM sodium pyruvate and 50 µM 1-thioglycerole  in the absence of further cytokines as described The IVIS Lumina II Imaging System was used . Mice were anesthetized using isoflurane, placed into the imaging chamber in a supine position and fixed at the lower limbs and by the inhalation tube. Coelenterazine  was dissolved in acidified methanol (HPLC grade) at concentration 10 mg/ml and diluted shortly before injection in sterile HBG buffer . Immediately after intravenous tail vein injection of 100 µg of native Coelenterazine, mice were imaged for 15 seconds using a field of view of 12,5 cm with binning 8, f/stop 1 and open filter setting. To monitor tumor growth, mice were imaged once weekly; after therapy, mice were imaged every other day.4 photons per second per cm2 per solid angle of one steradian (sr). Different regions of interest (ROI) were defined and signals were considered positive, when light emission exceeded background in each ROI; background was measured in 15 mice harboring GLuc negative leukemias; a ROI covering the entire animal was used (background 4×106 photons per second); as an exception and to determine early engraftment = minimal disease, a small ROI  was set at femurs at the location, where and when first light emission became visible; depending on the expression level of the transgenes, overt leukemia was considered above 109–1010 photons per second using the ROI covering the entire animal; overt leukemia served as criterion for ending experiments, as it shortly preceded onset of clinical signs of disease in mice.The Living Image software 4.x  was used for data acquisition and quantification of light emission using a scale with a minimum of 1,8×10Control animals received physiological salt solution intraperitoneally; treatment group mice were injected i.p. with a single dose of either Etoposid  or Cyclophosphamide  diluted in 0.9% NaCl.http://bioinf.wehi.edu.au/software/elda).Mean and standard error of the mean (SEM) were calculated using the Microsoft Excel 2010 software . To determine significance of treatment effects in vivo, Mann-Whitney Rank Sum Test and the Sigma Plot 12 software  was used. CSC frequencies were calculated according to Poisson statistics using the ELDA software application  was chosen in its membrane-bound form Leukemia cells are notoriously difficult to transfect and patient-derived leukemia cells do not allow antibiotics-based selection in vitro. Therefore, lentiviral transduction was chosen, although transgene integration into unsuitable genomic sites might alter cell function. GLuc was cloned into a lentiviral vector harboring additionally copepod green fluorescence protein (GFP) . PrimaryAll 9 samples from children with ALL  were successfully transduced, although transduction efficiencies and levels of transgene expression varied widely between samples . While hFor imaging, the convenient IVIS Lumina II Imaging System  was used together with an optimized protocol (for details see suppl. Results). Kinetics of GLuc-emitted light from leukemia cells was similar to published kinetics on GLuc-expressing T-cells Bioluminescence in vivo imaging using GLuc visualized engraftment of patient-derived ALL cells in mice first in the lower extremities where bones and bone marrow are located directly under the fur . Over tiLikewise, light emission strongly depended on the number of cells injected, indicating that both kinetic and dose response were clearly represented by in vivo imaging in this model . The groTo reassure that imaging reliably visualized the leukemic disease, in vivo imaging data were correlated to conventional post mortem readouts. Single mice engrafted with sample ALL-50 were sacrificed weekly and bone marrow and spleen were analyzed by flow cytometry and immunohistochemistry to quantify leukemic cells. Imaging was more sensitive in detecting leukemic infiltration in bone marrow than in spleen arguing towards the highest sensitivity of imaging in visualizing ALL in bone marrow located directly under the fur , but wasTo quantify light emission, regions of interest were defined and analyzed using the Living Image software 4.0 . In close correlation to the visual impression, quantification revealed first engraftment in a region covering the lower extremities, while inner organs became visible at late time points followed by extremely rapid signal increase . For furQuantification of light emission revealed a strictly logarithmic growth of patient-derived leukemia cells in mice covering up to 4 orders of magnitude . SimilarIndividual leukemias were obtained from patients with very different clinical parameters, including primary disease and relapse, and contained completely different cytogenetic and molecular alterations . Light emission quantification of ALL samples showed that all samples grew in a logarithmic pattern in mice . Thus, logarithmic growth appears as general principle how human leukemia cells behave in mice. Nevertheless and according to published data To control quality parameters of the model, assay variances were estimated. Imaging showed a highly reproducible growth of leukemia in mice with surprisingly low intra- and inter-assay variances considering that an in vivo model was studied  represents a major clinical threat as MRD is difficult to treat and often followed by relapse. Appropriate preclinical models to study MRD are required 4 photons per second using defined criteria, see Methods for details), most mice were sacrificed and bone marrow and peripheral blood analyzed by flow cytometry, quantitative real time PCR and immunohistochemistry. All remaining mice showed constant light increase similar to the kinetic shown in To test, whether GLuc-based in vivo imaging could visualize MRD, its sensitivity was measured. Groups of mice were engrafted with equal cell numbers of the same sample and were imaged three times weekly. Upon crossing a clearly defined detection threshold  are able to maintain tumor growth and therefore represent the most important targets for anti-cancer therapy Bioluminescence in vivo imaging allowed observing each group of mice within the LDTA repetitively over time . TherebyTNF-related, apoptosis-inducing ligand) prior to transplantation significantly reduced engraftment of cells in mice . Imaging allowed starting treatment in mice with equal tumor burdens, although rarely mice had to be excluded, as leukemia grew highly homogenously in mice . Mice were treated with conventional cytotoxic drugs by systemic bolus injections in concentrations modeling drug doses typically applied in patients Individual leukemias were grown until advanced disease . Treatment nearly eliminated tumor cells from peripheral blood, while imaging visualized the remaining tumor burden post treatment . Due to limited sensitivity, conventional measurement in peripheral blood appears overestimating therapeutic effects, although this readout was most frequently used for disease monitoring until today.In our model, each mouse harbors the leukemia of an individual patient. Imaging-guided preclinical treatment trials in mice revealed that individual ALL samples retained individual sensitivities towards conventional cytotoxic drugs. While the T-ALL sample ALL-4S was sensitive towards treatment with Cyclophosphamide, it was resistant towards Etoposid; in contrast, the B-ALL sample ALL-199 was resistant towards Cyclophosphamide, but sensitive towards Etoposid . AccordiMost importantly, imaging was able to quantify therapeutic responses and to visualize tumor regrowth. Due to low assay variances, imaging allowed distinguishing treatment failure/drug resistance/progressive disease from partial or complete response . As addiTaken together, in vivo imaging allowed the rapid, precise and individual quantification of treatment responses.The sensitivity of tumor cells towards treatment depends on the disease stage at which the drug is given After treatment, imaging reliably quantified residual disease. Short-interval imaging revealed short “lag phase” of no-growth after a single application for certain, but not all drugs and samples . In the Taken together, we have introduced and validated in detail GLuc-based bioluminescence in vivo imaging in the xenograft mouse model of individual patient’s ALL. Imaging was easy to perform and gave rise to highly sensitive and reliable results which enable the non-invasive accurate and detailed monitoring of disease progression and treatment responses.GLuc-based bioluminescence in vivo imaging was introduced and intensively validated as novel readout parameter for the preclinical model of patient-derived ALL growing in mice. In vivo imaging allowed performing preclinical trials on a novel level of accuracy and precision including stage-specific therapy and quantification of treatment responses. In the future, improvement of the individualized ALL mouse model by in vivo imaging will allow performing preclinical trials more exactly and in more detail.Imaging allowed modeling disease stages in mice which represent current challenges in the clinics such as minimal residual disease and tumor regrowth. Imaging was highly sensitive and continuous and correlated well with post mortem results regarding tumor distribution. Assay variances were minimal which will reduce the number of animals required per experiment. In addition, tumor growth was orthotopic and homogenous and tumor cells were derived from individual patients with genetically defined tumors; limiting dilution assays were easily visualized to study drug sensitivity of leukemia initiating cells. Taken together, GLuc-based imaging will allow performing high quality and convenient preclinical treatment trials in the individual mouse model of ALL in the future.The most important challenges in cancer treatment represent treatment failure and relapse. Using in vivo imaging, treatment failure is easily detectable in the first days after treatment as light emission continues increasing despite of therapy; short-period treatment quantification will allow speeding up preclinical treatment trials. Tumor regrowth is visualized by quantification of post-therapeutic residual disease followed by monitoring increase in light emission. Thus, our model is able to exactly map both important clinical challenges for preclinical trials.In addition to bioluminescence imaging, fluorescence imaging represents an interesting alternative with a better anatomical resolution, especially using near- infrared fluorochromes Recently, two groups published the use of firefly luciferase for in vivo imaging of transplanted individual ALLs The major advantage of using GLuc instead of firefly luciferase for in vivo imaging is the markedly increased light emission from superficial organs such as bone marrow in the lower extremities of mice Leukemic disease serves as a suitable model disease for cancer in general since leukemia cells are easier in handling compared to solid tumor cells. Many important research discoveries in cancer biology were first described in leukemia; for example, oncogenic mutations are best characterized in acute myeloid leukemia Taken together, GLuc-based in vivo imaging in the individualized preclinical model of ALL enables performing treatment trials on a novel level of accuracy and precision. It enables quantifying therapy effects and remaining disease burdens as well as exact modeling of distinct disease stages. The model facilitates the detailed preclinical analysis of novel therapies for preparing their translation into the clinics. The model allows preclinical trials addressing the most demanding current clinical challenges, such as treatment failure, resistance in leukemia initiating cells, minimal residual disease and relapse.Supporting Information S1(PDF)Click here for additional data file.


The link between high maternal blood pressure and poor pregnancy outcome is well established. Similarly the causal relationship between acute maternal hypotension and acute fetal distress is well recognised. The link between poor pregnancy outcome and persistent maternal hypotension is less well known.McClure Brown  was the It is problematic that previously reported studies have defined maternal hypotension differently. However, Warland  attempteIn summary, there is a small body of research which has consistently demonstrated the negative effect of persistent maternal hypotension on poor pregnancy outcome including stillbirth. These studies have been conducted using a range of approaches including prospective cohort, retrospective case-control and population based data bank analysis. Many questions remain unanswered, including the definition of hypotension, the level at which hypotension becomes problematic, and how best to manage maternal hypotension in pregnancy.


BCR-ABL translates into an abnormal tyrosine kinase that accelerates development of chronic myelogenous leukemia [Gene fusions are hybrid genes formed when two discrete genes are incorrectly joined together. Gene fusions are found to play roles in tumorigenesis. For example, the fusion gene leukemia . A netwoleukemia  performeWe mined three public databases for cancer-related gene fusion sequences, and one database for fusions records from cancer studies, transcriptome analysis, and genetic disorders. Specifically, we processed each dataset by removing incomplete entries and then extracted gene-fusion pairs. Genes serve as the nodes in the network and each fusion pair is joined by an edge. Repeating pairs were represented once. We used Cytoscape to build five networks: one for each dataset and the fifth that encompasses all datasets. We graphed the occurrence of degree in each single dataset network to determine an empirical definition for hub genes.MLL and MALAT1, both of which have roles in tumorigenesis. The network also highlights genes such as WDR74 and COL1A1, which are not much studied.The comprehensive network includes 9852 genes, displays 12,791 relationships, and highlights 1248 hub genes. The network highlights genes such as This preliminary network analysis provides interesting features of tumorigenesis-related fusions. Further systematic analysis of gene fusion networks may aid researchers to better understand cancer gene fusions and test novel fusions in specific types of cancer.


Thirty-four countries worldwide have abnormally high sex ratios (>102 men per 100 women), resulting in over 100 million missing women. Widespread sex selective abortion, neglect of young girls leading to premature mortality, and gendered migration have contributed to these persistent and increasing distortions. Abnormally high adult sex ratios in communities may drive sexually transmitted disease (STD) spread where women are missing and men cannot find stable partners. We systematically reviewed evidence on the association between high community sex ratios and individual sexual behaviors.Seven databases  were searched without restrictions on time or location. We followed PRISMA guidelines and evaluated quality according to STROBE criteria. 1093 citations were identified and six studies describing 57,054 individuals were included for review. All six studies showed an association between high community sex ratios and individual sexual risk behaviors. In high sex ratio communities, women were more likely to have multiple sex partners and men were more likely to delay first sexual intercourse and purchase sex. Only two studies included STD outcomes.High community sex ratios were associated with increased individual sexual risk behavior among both men and women. However, none of the studies examined unprotected sex or appropriately adjusted for gendered migration. Further studies are needed to understand the effect of community sex ratios on sexual health and to inform comprehensive STD control interventions. Sex selective abortions have become so widespread that the global sex ratio at birth has increased from 105 to 107 High sex ratios establish communities where surplus men cannot form stable partnerships with women, potentially driving risky sexual behaviors that accelerate transmission of sexually transmitted diseases (STDs). Increased STD burden in high sex ratio communities may be due to increased unsafe commercial sex The potential for high adult sex ratios to drive STD transmission has been previously hypothesized surplus men, forced bachelor, sex ratio, male-female ratio, or gender ratio, and HIV/AIDS, std, sexual behavior, sexually transmitted disease, risk behavior, sex workers, or commercial sex. Our PubMed search terms are available in We conducted a literature search through 30 August 2012 of articles that addressed the association between adult community sex ratios and individual sexual risk behaviors and STD biomarkers. We searched abstracts using combinations of the key words Abstracts were checked for potential relevance, and had to meet the following criteria: only quantitative, English-language population-based research studies; sex ratios measured across multiple partner markets (a community unit defined by each study); and reported individual sexual risk outcomes (behaviors or biomarkers). In order to conduct a comprehensive global systematic review, no limits were placed on study date, location, race or ethnicity of study participants, or the definition of sex ratio. Partner markets were defined as discrete geographic units in which individuals were more likely to find a stable partner. Behavioral outcomes included in the study were multiple sex partners, sex with commercial sex workers, forced sex, premarital sex, recent sex, and self-reported sexually transmitted disease. Biomarker outcomes included a positive test for any sexually transmitted disease. We excluded studies conducted in settings with primarily low sex ratios  and studies that were not population-based. Outcomes related to health seeking behaviors, mental health, or unrelated to sexual behavior were excluded. Case studies, qualitative studies, and ecological studies were excluded.We used PRISMA guidelines to identify and exclude studies (Text S2). Two independent reviewers (CB and JT) analyzed full-text articles for inclusion in the review. The reference sections of the remaining articles were then searched to identify other studies that met our inclusion criteria. Source data were reviewed (by JT and CY) and data were abstracted into tables. We contacted study authors to retrieve missing study data. Effect size was measured according to parent data, as either the coefficient or odds ratio. In some cases, statistical significance was converted to a p-value. We used adjusted odds ratios in studies with both crude and adjusted data.We graded the quality of each study using the STROBE reporting criteria for cross-sectional studies  [30]. WeOur search yielded a total of 1093 studies . Most inSix studies were included in this review describing 57,054 individuals. Three studies were conducted in high-income countries Definitions of adult sex ratios, migration, and partner markets substantially differed between studies. Although the adult sex ratio is typically reported as the number of men per 100 women of a certain age, two studies reported the number of women per 100 men. These measures were not converted into standard measures of sex ratios in order to preserve apparent effect sizes from the studies. None of the studies included individual-level data for migration, although two of the studies included migrants in community-level census data but excluded migrants in the study sample All six studies found an association between high sex ratios and increased sexual risk outcomes . Among tTwo studies found a concurrent decrease in some sexual risk outcomes and increase in other sexual risk outcomes Few of these studies examined mechanisms linking the connection between high community sex ratio and individual behavior. Two studies examined purchasing of sex among men High adult sex ratios are increasingly common in many parts of the world, but the impact of these global demographic changes on individual sexual behaviors is poorly understood. The proportion of men older than 25 years old who fail to marry in China will nearly triple in the next twenty years, even if sex ratio trends reverse now Our review found limited evidence that high sex ratios were associated with increased sexual risk behavior among women. One study in China Our review suggests increased sexual risk behaviors among men in high sex ratio communities as well. Two studies found that men in high sex ratio communities delay first sexual intercourse, but have an increased risk of purchasing sex, compared to men in normal sex ratio communities Understanding the mechanisms that link community sex ratios and individual risk can guide risk-reduction strategies, from community-level to individual-level interventions. However, we found little evidence that linked the impact of community sex ratios and individual sexual risk behaviors . The onlMigration of surplus men away from high sex ratios is a critical issue because it may inadvertently increase sex ratios in destination communities. However, none of the studies examined gendered migration as either a potential driver of increased sex ratios or a potential cause of individual risk behavior. One study of migrant communities in North Carolina with high sex ratios found an increase in commercial sex use among in communities with fewer women Despite our comprehensive search strategy, our review highlights the relatively limited literature on community sex ratios and sexual health. The reporting quality of some of the studies was poor. One study presented only data that did not differentiate male and female sexual behaviors Our review has several limitations. First, meta-analysis was not possible due to substantial variation in study methodology. Although sex ratios are commonly reported as number of males per 100 females, two studies reported sex ratios as the number of females per 100 males. Variations in study definition of sex ratio and outcomes measures did not permit comparisons of effect size. In order to broaden the scope of the review, we did not place any restrictions on time or location of included studies. Nevertheless, our study included data from only four countries, introducing the possibility of publication and location bias. Three of the included studies were conducted in countries with high sex ratios, but other nations, particularly those in Asia, also have abnormally high sex ratios. Future studies in these settings are warranted. Increased understanding of the mechanisms that drive sexual risk behavior in high sex ratio communities will strengthen comparability between study settings. We did not include studies where sex ratios are predominately low, a demographic phenomenon noted in African-American communities, communities that experience net out-migration, and nations in Eastern Europe Profound demographic changes will fundamentally shift population structures across the globe. Sex ratios will continue to rise in many regions of the world, but their effect on human health, particularly sexual health, remains poorly described. Our review found evidence that high community sex ratios may be associated with increased sexual risk behaviors among both men and women. More epidemiology research, especially among men, is needed to understand the mechanisms driving this association. In addition, prospective studies that include individual STD and migration data are warranted to further assess the findings in this review. Understanding the impact of community sex ratios on sexual health will inform structural STD control interventions, which can better target populations at increased STD risk.Table S1STROBE reporting criteria for cross-sectional studies (full-text).(DOCX)Click here for additional data file.Text S1PubMed Search Strategy.(DOCX)Click here for additional data file.Checklist S1PRISMA Checklist.(DOC)Click here for additional data file.


Arabidopsis veins provide an experimental system to identify such switches. The extent of ingrowth deposition responds to various abiotic and applied stresses, enabling bioinformatics to identify candidate regulatory genes. Furthermore, simple fluorescence staining of PP TCs in leaves enables phenotypic analysis of relevant mutants. Combining these approaches resulted in the identification of GIGANTEA as a regulatory component in the pathway controlling wall ingrowth development in PP TCs. Further utilization of this approach has identified two NAC -domain and two MYB-related genes as putative transcriptional switches regulating wall ingrowth deposition in these cells.In species performing apoplasmic loading, phloem cells adjacent to sieve elements often develop into transfer cells (TCs) with wall ingrowths. The highly invaginated wall ingrowths serve to amplify plasma membrane surface area to achieve increased rates of apoplasmic transport, and may also serve as physical barriers to deter pathogen invasion. Wall ingrowth formation in TCs therefore plays an important role in phloem biology, however, the transcriptional switches regulating the deposition of this unique example of highly localized wall building remain unknown. Phloem parenchyma (PP) TCs in  These cells inuities . The incArabidopsis  often develop extensive wall ingrowths. Well-known examples include companion cells (CCs) in pea , phloem bidopsis , and botvulgaris . In pea,vulgaris , and in e PP TCs . TCs aree PP TCs , particue PP TCs . Wall inVicia faba cotyledons have established that auxin (V. faba cotyledons (Arabidopsis that enabled a combined bioinformatics and reverse genetics approach to be undertaken to discover that GIGANTEA (GI)is a component of a pathway regulating wall ingrowth deposition in PP TCs. Further, we describe preliminary results using this approach to identify previously uncharacterized members of the NAC -domain and MYB-related gene families as putative transcriptional regulators of wall ingrowth deposition in PP TCs.Transfer cell development occurs across normal developmental windows but also in response to biotic and abiotic stress . Recent at auxin , ethylenat auxin , and reaat auxin  functiontyledons  and endotyledons , 2012 inArabidopsis are known to occur in PP of the minor vein network in both leaves . Higher magnification views revealed that the Calcofluor White staining showed a distinctive mottled appearance, a characteristic consistent with staining the patchy and tangled wall ingrowths seen in leaf PP TCs by scanning electron microscopy . Double labeling experiments have shown that Aniline Blue gives the same mottled patterns of staining for PP TCs as does Calcofluor White . The levels of reduced abundance in each line, while significant, were not comparable to that seen for the gi-2 mutant (Table 1), indicating the possibility that these transcription factors may be acting redundantly with unidentified orthologs in controlling wall ingrowth deposition. In silico expression data (eFP and Genevestigator) shows that all four genes are expressed at very low levels in leaves, and qPCR confirmed this observation directly for both expanding and fully expanded leaves . Low expression might be expected for genes operating as putative regulators of wall ingrowth deposition specifically in PP TCs, since the number of PP TCs relative to most other cell types in the leaf is exceedingly low (AtSWEET11 promoter) over-expression to test the role of these transcription factors as regulators of wall ingrowth deposition in Arabidopsis.Based on the successful approach used by ngly low , and manngly low . Given ttrans-differentiation of non-vascular cells into metaxylem- and protoxylem-like vessel elements, respectively  or VND7, both NAC-domain transcription factors, causes ectively , a proceectively  and AtMYectively  also cauectively . Buildinectively , thus ouectively , 2009.Arabidopsis has proven useful to identify candidate genes operating as putative transcriptional regulators of wall ingrowth deposition in TCs. The discovery of GI as a component in the pathway regulating wall ingrowth deposition, and identification of NAC-domain and MYB-related genes as putative “master switches” involved in controlling this process, provides new lines of investigation to understand the genetic control of TC development and the cell biology of localized wall ingrowth deposition. Ultimately, identifying master switches which respond to various inductive signals to coordinate wall ingrowth deposition in TCs may provide new opportunities for improving crop yield by manipulating this process.The formation of wall ingrowths in TCs impacts on phloem loading and post-phloem unloading processes in many species, with corresponding impacts on plant development and reproduction. Development of an experimental system to investigate PP TCs in 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.


Ciliopathies is an emerging class of genetic disorders due to altered cilia assembly, maintenance or function. Syndromic ciliopathies affecting bone development have been classified as skeletal ciliopathies. Mutations in genes encoding components of the intraflagellar transport (IFT) complex A, that drives retrograde ciliary transport, are a major cause of skeletal ciliopathies. Mainzer-Saldino syndrome (MSS) is a rare disorder characterized by phalangeal cone-shaped epiphyses, chronic renal failure and early-onset severe retinal dystrophy.IFT140 mutations in seven MSS families. The effect of the mutations on IFT140 localization was assessed using flagged-IFT140 mutant proteins which showed a partial to nearly complete loss of basal body localization associated with an increase of cytoplasm staining while the wild-type Flagged-IFT140 protein predominantly localized to the basal bodies in RPE1 cells. To assess the impact of IFT140 mutations on ciliogenesis, abundance and morphology of primary cilia were studied in cultured fibroblasts of patients and detected absent cilia in a high proportion of patient cells compared to controls. Ciliary localization of anterograde IFTs were altered in MSS patient fibroblasts supporting the pivotal role of IFT140 in proper development and function of ciliated cells.We collected 16 families presenting three diagnostic criteria of MSS. Through ciliome re-sequencing combined to Sanger sequencing, we identified IFT140 mutations in seven MSS families. After Sensenbrenner and Jeune syndromes, MSS is the ultimate skeletal ciliopathy ascribed to IFT disorganization.Here we report on compound heterozygosity or homozygosity for


Obsessive-compulsive disorder (OCD) is characterized by an excessive focus on upsetting or disturbing thoughts, feelings, and images that are internally-generated. Internally-focused thought processes are subserved by the “default mode network" (DMN), which has been found to be hyperactive in OCD during cognitive tasks. In healthy individuals, disengagement from internally-focused thought processes may rely on interactions between DMN and a fronto-parietal network (FPN) associated with external attention and task execution. Altered connectivity between FPN and DMN may contribute to the dysfunctional behavior and brain activity found in OCD.The current study examined interactions between FPN and DMN during rest in 30 patients with OCD (17 unmedicated) and 32 control subjects (17 unmedicated). Timecourses from seven fronto-parietal seeds were correlated across the whole brain and compared between groups.OCD patients exhibited altered connectivity between FPN seeds (primarily anterior insula) and several regions of DMN including posterior cingulate cortex, medial frontal cortex, posterior inferior parietal lobule, and parahippocampus. These differences were driven largely by a reduction of negative correlations among patients compared to controls. Patients also showed greater positive connectivity between FPN and regions outside DMN, including thalamus, lateral frontal cortex, and somatosensory/motor regions.OCD is associated with abnormal intrinsic functional connectivity between large-scale brain networks. Alteration of interactions between FPN and DMN at rest may contribute to aspects of the OCD phenotype, such as patients' inability to disengage from internally-generated scenarios and thoughts when performing everyday tasks requiring external attention. Obsessive-compulsive disorder (OCD) is characterized by intrusive thoughts, feelings, or images (obsessions) and repetitive behaviors (compulsions) aimed at reducing anxiety associated with obsessions. Neuroimaging studies examining brain activation in OCD at rest, during symptom provocation, and in response to cognitive tasks have made critical advances in elucidating the neurobiological substrates of the disorder, pointing to dysfunction in several cortical and subcortical regions. While dysfunction in orbital, medial frontal, and striatal areas composing fronto-striatal circuits A growing body of literature in neuroscience has begun to emphasize the importance of interactions between brain regions, due to the realization that a typical brain area is likely to support multiple cognitive functions and that unique functionality is most likely to emerge through inter-regional connectivity Recent investigations of rs-fcMRI in OCD focusing on the striatum The investigation of competitive interactions between FPN and DMN at rest is particularly relevant for the study of OCD. Not only are nodes of these networks found to be abnormal during task-based studies of OCD, but the phenomenology of the disorder is consistent with the idea of a disrupted relationship between ongoing internal thought and external information, in that patients often excessively focus on internally-generated fears that are inconsistent with evidence present in the external environment This research was approved by the Institutional Review Board of the University of Michigan Medical School, following the principles set forth by the Declaration of Helsinki. All subjects provided written informed consent. Resting-state functional connectivity data were acquired for a total of 69 subjects. Seven subjects were excluded due to technical problems , leaving a total of 62 participants including 30 OCD patients and 32 control subjects for further analysis. Seventeen OCD patients were unmedicated for a minimum of 6 months prior to study participation (uOCD) and 13 were medicated (mOCD), primarily with serotonin-reuptake inhibitors  without psychiatric diagnoses and 15 medicated patient controls (mPC). Subjects with any history of OCD were excluded from both control groups. Subjects in the mPC group were patients with remitted major depression who were on SRI medication  and had Subjects were evaluated by a trained clinician using the Structured Clinical Interview for DSM-IV . A T1-weighted image was acquired in the same prescription as functional images to facilitate co-registration. Functional images were acquired with a T2*-weighted, reverse spiral acquisition sequence  sensitive to signal in ventral frontal regions 3, normalized to the template MNI152 brain , and smoothed with a 5 mm isotropic Gaussian smoothing kernel using the Statistical Parametric Mapping (SPM) 2 package .Physiologic signals (heart rate and respiration) were removed from the data using RETROICOR www.nitrc.org/projects/conn, see ref. Analysis of functional connectivity of LFBFs was carried out with the “conn" toolbox  controlled for correlations due to movement. Data were filtered between .01 and .10 Hz. We examined connectivity patterns separately for seven different seed regions-of-interest (ROIs) located in FPN. Given abundant evidence of positive correlations among FPN nodes, the timecourses for these separate seeds are not likely to be completely orthogonal; thus we cannot make strong claims about distinct patterns of connectivity between the different seeds. Nevertheless, given recent evidence identifying dissociable cingulo-opercular and dorsolateral prefrontal and parietal systems within FPN Coordinates (in MNI format) for these seeds were taken from prior studies, which have identified a “core" task-set network that includes bilateral dorsal anterior insula  and posterior medial frontal cortex  Although not our main focus of analysis, we also sought to investigate within-DMN patterns of connectivity, due to two prior reports of reduced connectivity in OCD Relationships with symptom severity were examined by extracting connectivity values from regions showing group differences and correlating these with Y-BOCS scores  in the OCD group.Several post-hoc analyses were performed to examine the impact of other variables on results  and adjacent regions of posterior temporal cortex, and dorsomedial prefrontal cortex (DMPFC) . PatientHyper-connectivity in OCD was also found with dorsolateral and anterior inferior parietal seeds of FPN, although differences with DMN were less robust than with anterior insula seeds. For the left dorsolateral prefrontal (DLPFC) seed, patients had greater connectivity with right anterior insula/frontal operculum – also a part of FPN – as well as ventral occipital lobe/cerebellum. For the right DLPFC seed, patients not only had greater connectivity with the posterior cingulate cortex in the DMN, but also showed increased coupling with right lateral frontal regions of FPN.Fewer group differences emerged for connectivity with anterior inferior parietal seeds, with the left hemisphere exhibiting more connectivity with left pre-postcentral gyrus and the right hemisphere exhibiting more connectivity with PCC/precuneus for OCD patients as compared to controls.There were no regions within DMN that exhibited group differences in connectivity with anterior medial frontal (aMFC) or PCC seeds at the current threshold, which was corrected for multiple comparisons. However, as two prior studies have reported altered resting-state connectivity within DMN in OCD Connectivity between the right anterior insula seed and right thalamus was significantly related to Y-BOCS scores, with greater severity of symptoms associated with reduced connectivity . No other connectivity values were related to OC symptom severity.As is shown in the Supporting information , group dwithin DMN itself, although these effects only emerged at lower levels of significance. Negative connectivity among controls was evident despite the fact that global normalization was not used in the analysis Primary results from the current study revealed reduced negative connectivity between the fronto-parietal network and default mode network in OCD. Patients also showed more positive connectivity between fronto-parietal seeds and several areas outside DMN, including somato-motor areas  and other FPN regions . However, not all group differences reflected increased connectivity in OCD, as patients showed less positive connectivity than control subjects Research on DMN has garnered much interest since the initial observation of reduced blood flow within several anatomically-widespread brain regions during cognitive tasks versus passive viewing Recent evidence has identified dissociable sub-networks within FPN, with dorsal regions of anterior insula and medial frontal cortex forming a “core" network involved in implementing and maintaining attention to external task demands and detecting salient events Connectivity differences with FPN occurred across a number of DMN regions, including PCC, pIPL, DMPFC/aMFC, and parahippocampus. Despite the consistent activation of these brain regions across several different tasks involving internally-directed cognition OCD patients also showed greater positive interactions between FPN seeds and several areas outside of DMN, including pre-postcentral gyrus and posterior insula, which are part of a somatosensory/interoceptive and motor network Although the general absence of correlations with Y-BOCS scores might seem surprising, this may indicate that group differences reflect stable biomarkers of OCD not sensitive to symptom severity differences, similar to the mechanism suggested for the error-related negativity Unlike group differences found with FPN seeds, OCD patients showed less positive connectivity than controls subjects within DMN, a finding that was not due to differential motion. Although this effect was found only at a lowered threshold, it is consistent with two prior reports To our knowledge, this is the first report of altered intrinsic connectivity between distributed regions of fronto-parietal and default mode networks in OCD. However, there are several limitations of the current study, many of which could be addressed by future research. Given that results were not corrected for multiple seed comparisons, replications using a larger sample size are necessary. In addition, OCD and controls groups were not matched on generalized depression/anxiety and education levels. Although post-hoc inclusion of these factors in multiple regressions indicated that these effects were not driving the reported group differences in connectivity, future studies would benefit from investigating the effects of these variables on brain connectivity. Of particular interest for the study of psychopathology, hyperactivity in DMN has been identified in other psychiatric disorders Click here for additional data file.Table S1Medications taken by OCD patients (mOCD) and medicated control subjects (mPC). SSRIs = selective-serotonin reuptake inhibitors; SNRIs = serotonin-norepinephrine reuptake inhibitors; TCAs = tricycle antidepressants. All subjects except 1 mOCD patient were taking a serotonin reuptake inhibitor (SSRI or SNRI). Two mOCD and 3 mPC subjects were taking more than one medication .(DOCX)Click here for additional data file.Table S2Significance of diagnosis in predicting connectivity separately for unmedicated and medicated participants. Results obtained from multiple regressions also including generalized anxiety/depression and education as predictors. PCC = posterior cingulate cortex; pIPL = posterior inferior parietal lobule; DMPFC = dorsomedial prefrontal cortex; aMFC = anterior medial frontal cortex; aI/fO = anterior insula/frontal operculum.(DOCX)Click here for additional data file.


In children compliance with breath-hold instructions and motion control can be significantly reduced and there is limited data on CMR reproducibility.In adults CMR is the gold standard technique for evaluation of LV function, with reportedly excellent interstudy reproducibilityTo determine the interstudy reproducibility of cardiovascular magnetic resonance (CMR) measurements of left ventricular (LV) end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), ejection fraction (EF), cardiac output (CO), end-systolic mass (ESM) and end-diastolic mass (EDM) in children and examine the influence of analysis technique.CMR was performed in 10 healthy children aged 9 years as part of a study of developmental influences on cardiovascular structure and function. Contiguous short axis steady state free precession LV cine images were acquired. Scans were repeated following a short interval. Interstudy variability was assessed on datasets analysed using a manual post processing technique (Osirix). Each dataset was analysed with the papillary muscles and trabeculae included or excluded from the blood pool. Intra- and interobserver reproducibility was assessed on 10 datasets using both techniques.Inter-study coefficients of variation (CVs) were similar for measurements including or excluding the papillary muscles and trabeculae, though marginally smaller for the latter. CVs for measurements excluding the papillary muscles and trabeculae were generally higher than those reported from studies of adults . Nonetheless, for all parameters the variability between subjects was greater than the interstudy and interobserver variability combined (e.g. for CO standard deviations were 1.83 for between subject variability and 0.42 for interstudy variability respectively).CMR measurements of cardiac structure tend to be less reproducible in children than in adults, probably due to difficulty in following breath hold instructions. Nonetheless, between subjects variability is greater than interstudy and interobserver variability, indicating that useful measurements can be obtained for research studies.


Although social workers encounter many clients with substance use problems, curricula rarely require education on addictions. The screening, brief intervention, and referral to treatment (SBIRT) model was initially directed to practicing physicians. Recently, training has evolved to include medical students. This study describes the first known application of SBIRT training to social work students. The goal was to assess students’ knowledge of and attitudes toward alcohol misuse before and after SBIRT training. Students were given a questionnaire assessing attitudes, knowledge and perceived skills with regard to substance misuse. A computerized training session focused on symptoms of at-risk drinking and implementing SBIRT. Descriptive statistics explained overall knowledge, attitudes, and perceived screening and intervention skills. T-tests compared changes pre- and post-test. Seventy-four social work students (33 undergraduate and 41 graduate) completed the training modules and pre- and post-tests. Significant differences were found in seven of the 13 questions. Students reported more confidence in their ability to assess for alcohol misuse and successfully intervene with clients who have substance use behaviors. They reported feeling more strongly that routine screening and brief intervention were crucial to clinical practice. Incorporating alcohol screening and brief intervention techniques into social work practice is an important aspect of effective treatment. Training students to screen and intervene is critical to improving treatment skills. Teaching SBIRT is a simple and effective way to implement addictions education into social work curricula. Such training appears to increase students’ perceptions of their ability to change client behaviors and reduce client alcohol misuse.


This survey determined the extent to which the herb Moringa oleifera commonly used for medicinal and nutritional purposes is being consumed among HIV positive patients.The study was a cross-sectional survey carried out at Parirenyatwa Hospital Opportunistic Infections Clinic. A convenience sample of 263 HIV-infected adults was taken from the Zimbabwe National Antiretroviral Roll-out Program. Using a previously piloted researcher administered questionnaire; patients who reported to the clinic over six months were interviewed about their use of herbal medicines. The focus was on Moringa oleifera use, and included plant part, dosage, prescribers and the associated medical conditions.Sixty-eight percent (68%) of the study participants consumed Moringa oleifera. Of these, 81% had already commenced antiretroviral drugs. Friends or relatives were the most common source of a recommendation for use of the herb (69%). Most (80%) consumed Moringa oleifera to boost the immune system. The leaf powder was mainly used, either alone (41%) or in combination with the root and/or bark (37%).Moringa oleifera supplementation is common among HIV positive people. Because it is frequently prescribed by non-professionals and taken concomitantly with conventional medicine, it poses a potential risk for herb-drug interactions. Patient medication history taking should probe for herbal supplementation and appropriate counselling done. Further experimental investigations into its effect on drug metabolism and transport would be useful in improving the clinical outcome of HIV positive patients on HAART.


Stroke trial outcomes are usually classified in ordered functional categories, e.g. the modified Rankin Scale (mRS). Using a single cut-off to distinguish "good" and "poor" outcome is statistically inefficient and could miss effects in subgroups where few patients can be expected to achieve good outcome. "Ordinal shift analysis", using ordinal logistic regression to estimate a common odds ratio for all possible outcome category thresholds, is more efficient but could still obscure clinically important differences in response among prognostic subgroups. An alternative "prognosis based outcome" (PBO) approach defines separate, clinically meaningful good and poor outcome thresholds (GOTs and POTs) within each prognostic stratum. The strata are then combined in an overall trichotomous comparison of "good"/"intermediate"/"poor" outcomes.We defined prognostic categories, based on initial neurological scores, and derived suitable GOTs and POTs for the mRS, using data from a hospital stroke register. We then validated these thresholds using baseline and outcome data from the Scandinavian Candesartan Acute Stroke Trial (SCAST). Finally we performed trichotomous PBO analysis on the SCAST data.PBO analysis confirmed the overall neutral results of SCAST but did identify a notable negative treatment effect within the good prognosis group.This subgroup effect was not predicted, so requires independent confirmation. The PBO approach is clinically intuitive, can be used to analyse treatment response within different prognostic groups (as in cancer staging systems), and is statistically efficient. Also, it does not require multivariable modelling, making it easier to combine data from different studies in individual patient data meta-analysis.


Spinal lesions substantially impair ambulation, occur generally in young and otherwise healthy individuals, and result in devastating effects on quality of life. Restoration of locomotion after damage to the spinal cord is challenging because axons of the damaged neurons do not regenerate spontaneously. Body-weight-supported treadmill training (BWSTT) is a therapeutic approach in which a person with a spinal cord injury (SCI) steps on a motorized treadmill while some body weight is removed through an upper body harness. BWSTT improves temporal gait parameters, muscle activation patterns, and clinical outcome measures in persons with SCI. These changes are likely the result of reorganization that occurs simultaneously in supraspinal and spinal cord neural circuits. This paper will focus on the cortical control of human locomotion and motor output, spinal reflex circuits, and spinal interneuronal circuits and how corticospinal control is reorganized after locomotor training in people with SCI. Based on neurophysiological studies, it is apparent that corticospinal plasticity is involved in restoration of locomotion after training. However, the neural mechanisms underlying restoration of lost voluntary motor function are not well understood and translational neuroscience research is needed so patient-orientated rehabilitation protocols to be developed. Spinal cord injuries (SCIs) cause substantial social, economic, and health burdens. In the majority of cases, the spinal cord is not completely severed and thus some fiber tracts and segmental spinal cord circuits remain intact , which dIn addition to spontaneous reorganization of the brain after SCI, spinal cord circuitries have the capacity to alter their structure and function with motor training , as suppIn humans, BWSTT improves lower extremity motor scores, increases the amplitude of muscle activity in the ankle extensors during the stance phase of walking, and improves walking ability and clinical outcome measures –31. A reBased on the aforementioned findings, it is apparent that BWSTT contributes to restoration of locomotion. Because remodeling of neuronal circuits as a result of plasticity occurs at multiple sites of the central nervous system , 32 restThe corticospinal tract is the most direct pathway between the cerebral cortex and spinal cord with corticospinal axons monosynaptically synapsing onto spinal motor neurons. Even though neurons of the motor cortex are not required for simple locomotion, they exhibit a profound step-related frequency modulation in the cat –35. ThisThe involvement of supraspinal neural control in human walking can be assessed by a variety of techniques utilized in isolation or in combination, including electroencephalography (EEG), electromyography (EMG), transcranial magnetic and electric stimulation (TMS and TES), and neuroimaging , 46. SinA single stimulus of TMS produces a synchronized discharge of cortical interneurons and pyramidal neurons that travel down the corticospinal tract. Epidural electrodes in the spinal cord detect several waves following TMS, termed direct (D) and indirect (I) waves. I waves originate in the motor cortex most likely through activation of corticocortical projections onto corticospinal neurons , while Dα-motor neurons and interneurons [However, the MEP amplitude is not a reliable measure of corticospinal excitability. This is because TMS-induced action potentials in cortical axons spread transynaptically to many other neurons  that actrneurons .The aforementioned limitations can be counteracted by reducing the TMS intensity below the MEP threshold. Direct recordings in awake human subjects have shown that TMS at subthreshold MEP intensities, which does not evoke any descending corticospinal volleys, depresses the MEP evoked by a subsequent suprathreshold TMS  and the Various training protocols in uninjured subjects induce reorganization of corticospinal actions on lumbosacral motor neurons. For example, balance training decreased the TA and soleus MEP amplitudes , while 3In motor incomplete SCI subjects at rest, MEPs are either absent or very small in amplitude with prolonged latencies, which are considered signs of impaired transmission of the fastest conducting corticospinal neurons –77. FurtfMRI showed a greater activation in sensorimotor cortical and cerebellar regions following 36 BWSTT sessions [Reorganization of corticospinal actions with training in neurological disorders has been shown in few studies. In 4 male SCI subjects with tetraparesis, sessions  consistesessions . Three-tsessions . Furthersessions . The lowsessions .One person  with an American Spinal Injury Association (ASIA) Impairment Scale (AIS) D at Thoracic 5–7 received 60 BWSTT sessions  with a robotic exoskeleton device (Lokomat). Before training, the patient stepped at 0.5 m/s with 50% body weight support (BWS), and after training the patient stepped at 0.89 m/s with 20% BWS. Electrophysiological tests, illustrated as a schema in The TA MEPs evoked at 1.3 TA MEP threshold during assisted stepping before and after training are shown in The spinal cord constitutes the final common pathway for segmental and supraspinal pathways underlying motor behavior. Electrical stimulation of a mixed peripheral nerve at low intensities activates primary (Ia) afferent axons which synapse in the spinal cord. Alpha motor neurons activated monosynaptically by Ia afferent volleys induce a synchronized reflex response known as the Hoffmann-(H-) reflex , which iCortical control of spinal reflex circuits has been extensively investigated in awake humans by means of TMS. Subthreshold TMS produces a short-latency inhibition on the soleus H-reflex followed by a period of facilitation , 87–89 wIn addition to the H-reflex, the TA long-latency (or M3) ankle stretch reflex was facilitated when the MEP arrived in the spinal cord at the same time . HoweverPersistent changes in H- or stretch reflex amplitudes may be regarded as signs of learning and plasticity as a result of training, which have been shown after various training protocols. For example, 30 min ankle cocontraction training decreased the ratio of maximal H-reflex versus maximal M wave (Hmax/Mmax) and improved motor performance defined as the difference between the maximum and minimum torque displacements within 1 min . The solNonetheless, the aforementioned changes in H-reflex amplitude can result from modifications of interneuronal circuits interposed in the spinal pathway or by changes on the strength of descending pathways, since the latter is potent regulator of spinal reflex circuits behavior –107. ThiLimited evidence exists on plastic changes of the cortical control of spinal reflexes after locomotor training in neurological disorders. Forty BWSTT sessions in 29 patients with incomplete SCI reestablished the TMS-induced long-latency soleus H-reflex facilitation with subjects at rest . It shouIn One of the spinal interneuronal circuits with paramount contribution to the neural control of movement is that of disynaptic reciprocal Ia inhibition. Reciprocal inhibition refers to an automatic antagonist motor neuron inhibition when an agonist muscle contracts. Following an SCI, the reciprocal inhibition is either reduced or replaced by reciprocal facilitation –124 leadRegulation of locomotion by reflexly mediated spinal circuits that integrate sensory inputs is well established. The contribution of muscle afferents mediating information about the amplitude and rate of muscle stretch is easily recognized by the phase-dependent modulation of short-latency spinal reflexes during walking. The short-latency soleus and quadriceps extensor reflexes  in humans are modulated in a way that promotes bipedal gait. The ankle stretch and soleus H-reflexes increase progressively from mid- to late stance in parallel with the soleus EMG activity and are significantly depressed or abolished during the swing phase of gait –115, 125α motor neurons coincided with activity of Ia inhibitory interneurons [The soleus H-reflex depression during the swing phase in humans has been partly ascribed to reciprocal Ia inhibition exerted from common peroneal nerve group I afferents on soleus motor neurons, which is regulated in a similar manner to that reported in animals and corresponds largely to absent reciprocal inhibition in the stance phase and maximal in the swing phase , 128. Durneurons , 130. Iarneurons .Animal studies through intracellular recordings provided a detailed knowledge of the pathway and integration of segmental and supraspinal convergence at the interneuronal level –135 withDescending control of reciprocal inhibition has clearly been postulated in humans. In particular, the reciprocal inhibition exerted from common peroneal nerve group I afferents on soleus motor neurons was observed 50 ms before the onset of TA EMG activity . FurtherFindings on the reorganization of reciprocal inhibition as a result of motor training in health and disease are limited. Stimulation of the common peroneal nerve with a train of 10 pulses at 100 Hz with and without motor cortex stimulation potentiated reciprocal inhibition in control subjects . ReciproIn D-C)-(B-A) whereas A is the test soleus H-reflex (baseline soleus H-reflex modulation pattern during stepping), B is the soleus H-reflex conditioned by subthreshold TMS, C is the soleus H-reflex conditioned by common peroneal nerve stimulation , and D is the reciprocal inhibition conditioned by subthreshold TMS. Positive values indicate potentiation of reciprocal inhibition and negative values indicate attenuation of reciprocal inhibition. Locomotor training contributed significantly to attenuation of reciprocal inhibition exerted from ankle flexor afferents to extensor motor neurons during the stance phase. Most importantly, potentiation of reciprocal inhibition at swing phase initiation (i.e., bin 9 in The net effects of subthreshold TMS on the reciprocal inhibition during BWS-assisted stepping before and after 60 BWSTT sessions are indicated in SCI changes the human body homeostasis leading to myriad changes of multiple systems. In most cases, the spinal cord is not completely severed and thus some fiber tracts and segmental spinal cord circuits remain intact. Based on the plastic capabilities of the central nervous system, it is apparent that the adult lesioned motor system reorganization occurs spontaneously after an injury and after training. Electrophysiological studies have shown that BWSTT increases the MEP amplitude, changes the common drive of antagonist muscles from corticospinal inputs with subjects seated, and alters the TA MEP modulation pattern during BWS assisted stepping. Further, BWSTT reestablished the TMS-induced long-latency soleus H-reflex facilitation and potentiated the short-latency soleus H-reflex depression following subthreshold TMS with subjects at rest, while cortical modulation of the soleus H-reflex during stepping changed significantly. Lastly, BWSTT changed the cortical control of reciprocal inhibition during BWS assisted stepping in a manner that promotes bipedal gait. These findings support the notion that improvements in locomotor function from treadmill training are mediated, in part, by changes in the corticospinal drive of spinal reflex circuits, spinal interneuronal circuits, and output of leg muscles during walking.Plasticity in the brain and spinal cord underlying restoration of lost function can be driven by appropriately designed interventions , 151. De


These DNA modifications occur only during nuclear development and programmed genome rearrangement. We describe these modifications in three classes of eliminated DNA: germline-limited transposons and satellite repeats, aberrant DNA rearrangements, and DNA from the parental genome undergoing degradation. Methylation and hydroxymethylation generally occur on the same sequence elements, modifying cytosines in all sequence contexts. We show that the DNA methyltransferase-inhibiting drugs azacitidine and decitabine induce demethylation of both somatic and germline sequence elements during genome rearrangements, with consequent elevated levels of germline-limited repetitive elements in exconjugant cells. These data strongly support a functional link between cytosine DNA methylation/hydroxymethylation and genome stability, highlighting the ability of the cells to distinguish and specifically methylate aberrant chromosomes but not their properly structured isoforms. In addition, we identify a motif strongly enriched in some methylated/hydroxymethylated regions of parental MAC chromosomes, and we propose that this motif recruits DNA methylation machinery to specific chromosomal regions in the parental macronucleus. No recognizable methyltransferase enzyme has yet been described in O. trifallax, raising the possibility that it might employ a novel cytosine methylation machinery to mark DNA sequences for elimination during genome rearrangements.Cytosine methylation of DNA is conserved across eukaryotes and plays important functional roles regulating gene expression during differentiation and development in animals, plants and fungi. Hydroxymethylation was recently identified as another epi-genetic modification marking genes important for pluripotency in embryonic stem cells. Here we describe