S-nitrosothiols (SNOs), a class of endogenous airway smooth muscle relaxants. This deficiency results from increased activity of an enzyme that both reduces SNOs to ammonia and oxidizes formaldehyde to formic acid, a volatile carboxylic acid that is more easily detected in exhaled breath condensate (EBC) than SNOs. We therefore hypothesize that depletion of airway SNOs is related to asthma pathology, and breath formate concentration may be a proxy measure of SNO catabolism.Children with severe asthma have poor symptom control and elevated markers of airway oxidative and nitrosative stress. Paradoxically, they have decreased airway levels of 20 , and positively correlated with the NO-derived ion nitrite  as well as with total serum IgE . Furthermore, formate was not significantly correlated with other volatile organic acids nor with inhaled corticosteroid dose.We collected EBC samples from children and adolescents, including 38 with severe asthma, 46 with mild-to-moderate asthma and 16 healthy adolescent controls, and the concentration of ionic constituents was quantified using ion chromatography. The concentrations of EBC components with volatile conjugates were log-normally distributed. Formate was the principal ion that displayed a significant difference between asthma status classifications. The mean EBC formate concentration was 40% higher in samples collected from all asthmatics than from healthy controls . EBC formate was higher in severe asthmatics than in mild-to-moderate asthmatics . In addition, formate concentration was negatively correlated with methacholine PCS-nitrosothiols.We conclude that EBC formate concentration is significantly higher in the breath of children with asthma than in those without asthma. In addition, amongst asthmatics, formate is elevated in the breath of those with severe asthma compared to those with mild-to-moderate asthma. We suggest that this difference is related to asthma pathology and may be a product of increased catabolism of endogenous  Collection of bronchoalveolar lavage fluid and tracheal aspirate has previously been used to determine the concentration of This observational cross-sectional study was conducted based on the characterization procedures developed by the NIH/NHLBI Severe Asthma Research Program (SARP). The protocol was approved by the Emory University Institutional Review Board, and all procedures were monitored by an independent Data Safety Monitoring Board. The legal guardians of enrollees provided written informed consent, and the enrollees themselves gave assent when appropriate. All SARP participants met criteria for asthma as developed by the American Thoracic Society Guidelines.To compare the ionic composition of EBC from asthmatic children to that of non-asthmatic children, we analyzed EBC samples collected from adolescent athletes enrolled in a previous study. The features of this control group and study methods used have been previously described 1<80% predicted at baseline, more than one emergency department visit for asthma in the previous year, more than three oral corticosteroid bursts in the previous year, deterioration following a reduction in corticosteroid dose, or history of an asthma event requiring intubation). Participants who did not meet this definition of severe asthma were classified as having mild-to-moderate asthma.SARP enrollees were designated as having either severe or mild-to-moderate asthma based on the definition developed by the American Thoracic Society Consensus Group (ATS Consensus on Severe Asthma). This characterization procedure has been previously described in detail EBC was collected using the R-tube® . EBC pH was measured both before and after deaeration with argon. A separate aliquot was analyzed by ion chromatography using a previously described method 1 percent predicted, lower FEV1/FVC, and air trapping as indicated by a higher RV/TLC ratio. Although children with severe asthma exhibited mild airflow obstruction at baseline, FEV1 percent predicted increased significantly following maximum bronchodilation, and post-bronchodilator spirometric variables in children with severe asthma were not significantly different from those of children with mild-to-moderate asthma. Children with severe asthma were more atopic, with higher serum IgE and a greater number of positive skin prick tests to aeroallergens.The study sample included 38 children with severe asthma, and 46 children with mild-to-moderate asthma as well as 16 healthy adolescents enrolled in a previous study Table 1.Nearly all EBC samples contained quantifiable concentrations of the anions lactate, acetate, propionate, formate, butyrate, pyruvate, chloride, nitrite, nitrate, sulfate, and oxalate as well as the cations sodium, ammonium, potassium, magnesium, and calcium. The concentration of ammonium was approximately an order of magnitude greater than other measured ions.Principal component analysis identified three clusters of EBC constituents that account for most of the variance in ionic composition Figure 120  and serum IgE .Examination of the bivariate correlations between ions revealed a similar pattern: many non-volatile species were correlated with one other as were several organic species with a volatile conjugate, namely acetate, propionate, and butyrate. Ammonium was inversely correlated with these acidic species but did not otherwise exhibit a relationship with any other constituent. In addition, formate was significantly correlated with two clinical variables (not measured in controls), namely the methacholine PCThe key differences in EBC ionic composition between asthma severity classifications were found to be formate and to a lesser extent, nitrite concentration. The mean EBC formate concentration was 40% higher in samples collected from all asthmatics than from healthy controls . In addition, EBC formate was significantly higher in severe asthmatics than in mild-to-moderate asthmatics Figure 220 and serum IgE. Given the strength of the significant difference in EBC formate between asthma severity classifications, we suggest that the higher formate seen in the breath condensate of severe asthmatics may be related to asthma pathology.Of the ionic components measured in EBC collected from asthmatic and healthy children and adolescents, formate displayed the most notable difference between asthma severity classifications. Principal component analysis suggests that the source of formate in EBC is distinct from most other measured ionic species. Despite its high volatility in the non-ionized state, formate is not highly correlated with other volatile organic acids commonly detected in EBC. In addition, EBC formate is associated with the NO-derived ion nitrite, which itself displays more modest stratification by asthma severity. Furthermore, EBC formate was significantly correlated with clinical variables associated with asthma severity, namely methacholine PCS-nitrosothiols (SNOs). SNOs are a novel class of endogenous bronchodilators 2 adrenoreceptor. The principal SNO in healthy human airways is S-nitrosoglutathione (GSNO) S-hydroxymethylglutathione to form S-formylglutathione S-formylglutathione is in turn hydrolyzed to glutathione and formate A potential mechanism that conceptually explains both elevated breath formate levels and their association with asthma severity involves the catabolism of et al.S-nitrosothiol content and methacholine PC20et al. S-nitrosylation of G-protein-coupled receptor kinase 2 and that GSNOR deficient mice are protected from tachyphylaxis to β2 agonists. We therefore hypothesize that SNO degradation perturbs airway tone in asthma and suggest that GSNO may play an important role in future therapeutic strategies for the treatment of asthma.A growing body of evidence suggests that GSNO dysregulation is intimately involved in the pathology of asthma. Analysis of fluids obtained from the airways of children with respiratory failure and severe asthma have been found to have lower levels of SNOs relative to healthy children S-nitrosothiol bond is notoriously labile, introducing the potential for false positives and negatives S-nitrosothiol catabolism and is easily detectable as formate in EBC with no sample preparation needed S-hydroxymethylglutathione substrate is related to formaldehyde exposure Unfortunately, the concentrations of SNOs and GSNOR in human airways have proven difficult to evaluate. The


Against a backdrop of increasing antibiotic resistance, and the emergence of new and evolving pathogens, clinicians are increasingly forced to consider alternative therapies - probiotics are one such alternative. With life-cycles measured in minutes as opposed to years, bacteria have an extraordinary ability to evolve and adapt rapidly to changes in their environment . Thus, ivia a specific mode of action; for example, penicillin interferes with the cross-linking of two linear polymers by inhibiting the transpeptidase reaction, bacteriocins on the other hand have quite diverse activities. Nisin and many other structurally related lantibiotics for example, use the cell wall precursor lipid II bound to the membrane as a docking molecule for pore formation and combine at least two modes of action, i.e., pore formation and inhibition of cell wall biosynthesis, for antibacterial activity at nanomolar concentrations [Faced with an emerging pandemic of antibiotic resistance, clinicians and scientists alike are now struggling to find viable therapeutic alternatives to our failing antibiotic wonder drugs . One suctrations . These mtrations .However, despite their potent anti-pathogenic effect, a significant limitation of this approach is that probiotic bacteria tend to be physiologically fragile; often not surviving to sufficiently high numbers during prolonged storage in delivery matrices such as foods (yogurt and probiotic drinks) or tablet formulations . FurtherListeria monocytogenes in the probiotic strain Bifidobacterium breve, not only improves gastrointestinal colonisation and persistence, but also significantly bolsters the clinical efficacy of the probiotic strain [One approach to improving the physiological robustness and stress tolerance of probiotic strains is patho-biotechnology ,14. Essec strain .In addition to improving their physiological stress tolerance, resulting in improved delivery and persistence within the gut, recent studies have led to the development of 'designer probiotics' which specifically target enteric infections by blocking crucial ligand-receptor interactions between the pathogen and its target host cell ,18,19. Min vivo sensitivity to gastric acidity associated with oral application of therapeutic or prophylactic compounds, they can also be produced cheaply; grown to high levels, dried and stored for years at ambient temperatures.In addition to infection control Fig. , probiotIn conclusion then, "designer probiotics" can be engineered to kill pathogens, neutralise toxins, and facilitate re-colonisation of the resident beneficial microflora while at the same time priming both the innate and adaptive immune system; strengthening the host against subsequent infection - an approach far beyond the reach of conventional antibiotic therapies. Thus, the war against the antibiotic resistant "super bugs" may eventually be won by recruiting engineered "good bugs" as our allies.Bioengineered Bugs http://www.landesbioscience.com/journals/biobugs.Sleator is Editor-in-Chief of the peer reviewed scientific journal


Syndromic diarrhea (SD), also known as phenotypic diarrhea (PD) or tricho-hepato-enteric syndrome (THE), is a congenital enteropathy presenting with early-onset of severe diarrhea requiring parenteral nutrition (PN). To date, no epidemiological data are available. The estimated prevalence is approximately 1/300,000–400,000 live births in Western Europe. Ethnic origin does not appear to be associated with SD. Infants are born small for gestational age and present with facial dysmorphism including prominent forehead and cheeks, broad nasal root and hypertelorism. Hairs are woolly, easily removed and poorly pigmented. Severe and persistent diarrhea starts within the first 6 months of life (≤ 1 month in most cases) and is accompanied by severe malabsorption leading to early and relentless protein energy malnutrition with failure to thrive. Liver disease affects about half of patients with extensive fibrosis or cirrhosis. There is currently no specific biochemical profile, though a functional T-cell immune deficiency with defective antibody production was reported. Microscopic analysis of the hair show twisted hair (pili torti), aniso- and poilkilotrichosis, and trichorrhexis nodosa. Histopathological analysis of small intestine biopsy shows non-specific villous atrophy with low or no mononuclear cell infiltration of the lamina propria, and no specific histological abnormalities involving the epithelium. The etiology remains unknown. The frequent association of the disorder with parental consanguinity and/or affected siblings suggests a genetic origin with an autosomal recessive mode of transmission. Early management consists of total PN. Some infants have a rather milder phenotype with partial PN dependency or require only enteral feeding. Prognosis of this syndrome is poor, but most patients now survive, and about half of the patients may be weaned from PN at adolescence, but experience failure to thrive and final short stature.Syndromic diarrhea – Phenotypic diarrhea – Tricho-hepato-enteric syndrome – Intractable diarrhea of infancy with facial dysmorphism – Trichorrhexis nodosa and cirrhosis – Neonatal hemochromatosis phenotype with intractable diarrhea and hair abnormalities – Intractable infant diarrhea associated with phenotypic abnormalities and immune deficiency. To date, several types of early onset intractable diarrhea of infancy (IDI) have been recognized -8. Some Syndromic diarrhea (SD), also known as Phenotypic diarrhea (PD) or Tricho-hepato-enteric syndrome (THE), is a congenital enteropathy presenting with early-onset severe intractable diarrhea in infants born Small for Gestational Age (SGA) and associated with non-specific villous atrophy with low or no mononuclear cell infiltration of the lamina propria nor specific histological abnormalities involving the epithelium. The diarrhea is associated with facial dysmorphism, immune disorders and, in some patients, early onset of severe liver cirrhosis.et al. as unexplained diarrhea and failure to thrive in two siblings with unusual facies and abnormal scalp hair shafts [et al. in 1994 [et al.,1994 presented with an early-onset severe cholestatic disease that rapidly progressed to cirrhosis and death [SD is a newly described clinicopathologic entity with intractable diarrhea in infants. Two cases have been reported by Stankler r shafts . To date in 1994 -15. SomeSD appears to be much less common than microvillous atrophy ,7 or intThe patients present with diarrhea starting within the first 6 months of life (≤ 1 month in most cases). Severe malabsorption leads to early and severe protein energy malnutrition with failure to thrive and patients require parenteral nutrition (PN). Diarrhea persists while on bowel rest on PN. All affected infants have several features in common [Additional file From the reported cases in the literature, biopsies were performed during gastrointestinal endoscopy, at the time of referral to institutions or later at intervals depending on the therapeutic schedule. Biopsy specimens were stained with hematoxylin and eosin. Small intestine biopsies of the patients with SD show moderate Figure  or severet al. [In patients presenting with liver disease, pathological analysis of the liver usually shows macronodular cirrhosis with normal extra-hepatic ducts. Light microscopy examination shows extensive fibrosis or cirrhosis. Perl's staining shows iron depositions involving the hepatocytes and, to a lesser extent, the Kupfer cells. This aspect is consistent with neonatal hemochromatosis as suggested by Verloes et al. .Candida albicans, tetanus toxoid) and allogenic cell-induced lymphocyte proliferation. Serum immunoglobulin levels and specific antibody titers to poliovirus, tetanus, Diphteria toxoid and Bordetella pertussis were also measured in most reports. Functional T-cell immune deficiency with defective antibody production was described in the original report [in vitro. Further reports, in which extensive immunologic work-ups have been done, confirmed immune dysfunction. Several cases presented with monoclonal hyper IgA and/or hypo IgG [Published profiles have been studied by performing a complete immunologic work-up, including analysis of the T- and B- cell populations, and mitogen , antigen  suggests either mutations within several genes, inherited together by linkage disequilibrium, or, more probably, interference with a higher level of control, such as a patterning gene as seen in the Netherton syndrome which is an autosomal recessive congenital ichthyosis featuring chronic inflammation of the skin, hair anomalies, epidermal hyperplasia with an impaired epidermal barrier function, failure to thrive and atopic manifestations. The disease is caused by mutations in the SPINK5 gene encoding the serine proteinase inhibitor lympho-epithelial Kazal-type inhibitor [Among the congenital forms of hair dysplasia, tricorrhexis nodosa (TN) is very common and can be present in several pathologic conditions -19. TN iaciduria , citrullaciduria ,25, Menkaciduria , Nethertaciduria ,28, and aciduria . SD patinhibitor ,31. The The frequent association of the disorder with parental consanguinity and/or affected siblings suggests a genetic origin with an autosomal recessive transmission [Additional file Diagnosis may be suspected early from the clinical presentation with the association of the following anomalies:-intra-uterine growth retardation-severe protracted diarrhea of early onset-abnormal face with prominent forehead and cheeksMost patients also have abnormal hair with tricorrhexis nodosa, immune deficiency, long-term growth failure and mental retardation in common. Liver disease is associated in about half of the patients and is variable in severity.etc.) with rapid course to end-stage liver disease and patient death.Early management consists of total parenteral nutrition (TPN) using central venous catheter in an experienced clinical setting. Patients usually have persistent diarrhea and, in enterally fed patients, malabsorption is severe. Its mechanisms are unknown, as villous atrophy is usually not as severe as the diarrhea is, and small bowel bacterial overgrowth or specific malabsorption have never been documented in these patients. Long-term PN is required for achieving growth even if catch up growth, in patients born SGA, cannot be achieved. Attempts at enteral feeding should be performed in all cases using semi-elemental diet or amino-acid formulas. Some patients will tolerate progressive increase of enteral feeding making them able to be weaned from PN. However, normal growth velocity is not always achieved on full enteral feeding. Course may vary according to the concomitant liver disease, the severity of the digestive disease and the occurrence of infections. It is well reflected by the reported cases [Additional file Finally, the prognosis of this type of intractable diarrhea of infancy is poor, with >25% of the currently reported patients who died between the ages of 2 and 5 years, some of them with early onset of cirrhosis. In the largest but oldest series, five of the eight children reported died within 5 years due to sepsis or cirrhosis, despite aggressive intervention . More reThis very rare and heterogeneous syndrome has a poor prognosis with early death, liver cirrhosis or failure to thrive, and short stature with intellectual deficiency in the survivors. The heterogeneity of SD as well as the very different associated symptoms makes it very difficult to find an approach for understanding the genetic origin of the disease.Prognosis has improved since the description of the first case with SD. Most patients now survive and about half of the patients may be weaned from PN at adolescence, but experience failure to thrive and final short stature.Clinical features in patients with Syndromic (phenotypic) diarrhea. This table includes data from the published cases in the literature . Patients 18 to 25 are currently under publication by the authors.Click here for file


Beilstein Journal of Organic Chemistry a flavor of the synthetic challenges of liquid crystal research, as well as interdisciplinary aspects from material science, physics, physical and theoretical chemistry. It is a great pleasure to serve as a guest editor for this thematic series, where leading experts from all over the world have contributed original research papers and accounts that deal with diverse topics such as phthalocyanine-fullerene hybrid materials for photovoltaic devices, functional metallomesogens, coaxial electrospinning of liquid crystal-containing microfibers, banana-discotic hybrid systems, ionic liquid crystals, relations between molecular length distribution and formation of smectic phases, nematic phase engineering by using V-shaped molecules, saddle-shaped columnar systems displaying anomalous odd-even effects, theoretical studies on the origin of chirality transfer in liquid crystalline host-guest systems, liquid crystalline carboranes and dyes and discotic phenanthrene quinones. Many new exciting discoveries in this diverse research area are to be expected in the near future and our aim with this Thematic Series is to attract a new audience to this topic.Since their discovery in 1888 by the botanist Friedrich Reinitzer, who studied the melting behavior of cholesteryl benzoates, liquid crystals have developed from a mere curiosity to a highly interdisciplinary research field, with LC displays being the most important application to date . HoweverSabine LaschatGuest Editor


The existence of cancer stem cells would explain why only a small minority of cancer cells is capable of extensive proliferation of the tumor. In this review, we have discussed the studies on ovarian cancer stem cells along with the molecular pathways that could be involved in these cancer stem cells.Emerging evidence suggests that the capacity of a tumor to grow and propagate is dependent on a small subset of cells within a tumor, termed cancer stem cells. In fact, cancer cells, like stem cells, can proliferate indefinitely through a dysregulated cellular self-renewal capacity. Cancer stem cells may originate due to the distribution into self-renewal and differentiation pathways occurring in multi-potential stem cells, tissue-specific stem cells, progenitor cells and cancer cells. Recent studies have shown that ovarian cancer also contains stem cells or tumor-initiating cells. Moreover, ovarian serous adenocarcinomas were disaggregated and subjected to growth conditions to select for self-renewing, non-adherent spheroids previously shown to be derived from tissue stem cells. A recent study showed that epithelial ovarian cancer was derived from a sub population of CD44 It is the most lethal malignancy of the female reproductive system, at the initial stage the five-year survival rate is nearly 45%, which declines to 30% for patients with an advanced disease ,2. Great therapy . Cancer  therapy . The recStem cells, as classically defined, are cells with a capacity for self-renewal and generation of daughter cells that can differentiate into all the way down different cell lineages found in the mature tissue . Stem ceOf all the types of stem cell, hematopoietic stem cells (HSCs) are the best characterized adult stem cell . HSCs cain vivo tumor growth, whereas all other subsets  should not. Indeed, this assumption has now been repeatedly confirmed in several tumor systems. Three key observations classically define the existence of a CSC population: (i) Only the minority of cancer cells within each tumor are usually endowed with tumorigenic potential when transplanted into immunodeficient mice; (ii) Tumorigenic cancer cells are characterized by a distinctive profile of surface markers and can be differentially and reproducibly isolated from non-tumorigenic ones by flow cytometry or other immunoselection procedures; and (iii) Tumors grown from tumorigenic cells contain mixed populations of tumorigenic and non-tumorigenic cancer cells, thus recreating the full phenotypic heterogeneity of the parent tumor [Cancer stem cells may arise following transforming mutations that occur in untransformed stem cells, progenitor cells, mature cells, and cancer cells. The genetic program controlling self-renewal and differentiation plays a key role in the genesis of cancer stem cells Figure . Cancer nt tumor . Furthernt tumor ,30. In tThe identification of a reservoir of stem cells within many adult tissues raises the interesting possibility that all adult tissues have stem cells. Stem cell populations within normal tissues are defined by certain common characteristics: self-renewal to maintain the stem cell pool over time; regulation of stem cell number through a strict balance between cell proliferation, cell differentiation and cell death; and the ability to give rise to a broad range of differentiated cells ,32. It iin vivo. In this model, only a subset of cancer cells had the ability to form new tumors [low lineage cell population [et al. reported that a highly tumorigenic subpopulation of pancreatic cancer cells expresses the cell surface markers CD44, CD24 and epithelial-specific antigen (ESA) [To assay the cancer stem cells, a xenograft model for breast cancer was developed that allowed specific cancer tumors isolated directly from a patient to be passaged reliably w tumors . The canpulation . Furtherpulation ,7. In adpulation , only then (ESA) . Table 1en (ESA) .The field of stem cell research has given new hope for the treatment and even a cure for incurable diseases in human. Particularly, the identification of a rare population of adult stem cells in most tissues/organs in humans has emerged as an attractive source of multiple stem/progenitor cells for cell replacement-based therapies and tissue engineering in regenerative medicine. Our recent review discussed that cancer stem/progenitor cell research also offers the possibility of targeting these undifferentiated and malignant cells that provide critical function in cancer initiation and relapse for treating patients diagnosed with advanced and metastatic cancer ,35,36. VSince the cancer stem cells share common properties with normal stem cells, it is reasonable to think that they have overlapping regulatory mechanisms. Indeed, one of the most outstanding questions concerning the biology of stem cells is: how do multi-potent stem cells select a particular differentiation pathway and start to differentiate? Another question is how do stem cells decide to maintain self-renewal properties and continue to proliferate? Recent studies demonstrate that the presence of various genes and signaling pathways are involved in the regulation of the aforementioned processes. Among these, the Sonic Hedgehog (Shh), Notch and Wnt signaling transduction pathways play a major role in the self-renewal of stem cells -41. ReceThe ovaries contain three main types of cells germ cells, stromal cells and epithelial cells which give rise to germ cell, stromal and epithelial ovarian tumors, respectively. Epithelial ovarian cancers (EOC) were the most common type of ovarian cancers. Comprising nearly 90% of all ovarian cancers EOCs are derived from relatively pluripotent cells of the celomic epithelium or "modified mesothelium". These cells originate from the primitive mesoderm and can undergo metaplasia. Approximately 10% to 20% of epithelial ovarian neoplasms are borderline or low malignant potential tumors and are characterized by a high degree of cellular proliferation in the absence of stromal invasion. Of the invasive epithelial ovarian cancers, about 55–60% are serous, 15% endometrioid, 5–10% clear cell and <5% mucinous  Figure . The varIn general, tumor markers can be used for one of four purposes: (i) screening a healthy population or a high risk population for the presence of cancer; (ii) making a diagnosis of cancer or of a specific type of cancer; (iii) determining the prognosis of a patient; and (iv) monitoring the course in a patient in remission or while receiving surgery, radiation, or chemotherapy.et al reported that MUC1 molecule is not only expressed on mature cancer cells, but also on tumor cells that have multiple characteristics of stem and progenitor cells [+ tumors in vivo and that these tumors maintain a small population of MUC1+ cells with the stem/progenitor characteristics [Furthermore, recent studies have identified different prognostic and diagnostic surface markers for ovarian cancer  and thesor cells . This steristics . Our receristics . In addieristics . The futA recent study describes that ovarian cancer cell lines were shown to possess "side population" (SP) cells that have been described as cancer stem cells due to their stem-like characteristics including the ability to differentiate into tumors with different histologies. These putative cancer stem cells reflect the various histological subtypes observed in ovarian carcinoma. They also provide a model of cancer metastasis in which these cells are able to colonize, expand, and differentiate into heterogeneous tumor phenotypes similar to primary tumors. In such a model, both the primary tumors and metastasis would display similar genetic and expression profiles because both populations are supposedly derived from the same lineage of cancer stem cells . Ovarianin vivo. These cells are very organized self-renewing, anchorage-independent spheres and were reproducibly dividable using antibodies against both CD44 and CD117 [Furthermore, another study describes that two mouse ovarian cancer cell lines such as MOVCAR7 and 4306 contain candidate cancer stem cells . These tnd CD117 . These Ond CD117 , and thend CD117 ,60. One nd CD117 .The aforementioned studies showed that a so-called ovarian cancer stem cell, with high-proliferative capacity, self-renewal properties and multi-lineage potential, could be responsible for tumor development and the differentiation of more mature epithelial ovarian cells contributing to tumorigenesis. There are important consequences for cancer treatment if the growth of tumors is at least in part, dependent on a cancer stem cell population. The cancer stem cell hypothesis posits that cancer stem cells are a minor population of self-renewing cancer cells that fuel tumor growth and remain in patients after conventional therapy has been completed. The hypothesis predicts that effective tumor eradication will require obtaining agents that can target cancer stem cells while sparing normal stem cells. Experimental evidence suggests that ovarian cancer stem cells are relatively resistant to conventional chemotherapeutic agents. Current cancer therapies often engender severe toxicity because of their general effects on all rapidly dividing cells. Identification of candidate targets for more specific mechanism-based cancer therapy using techniques such as gene chips could reveal signature patterns of transcriptional output which are characteristic of activated self-renewal pathways.Emerging evidence suggests that these pathways also control patterning and growth in self-renewing adult tissues by regulating the stem-cell compartment. Thus, pharmacological inhibition of these pathways in the worst case might result in severe toxicity due to a loss of normal stem-cell compartments. Further research will be needed to determine whether continuous pathway activity is required in normal and tumor tissues, and whether these requirements differ sufficiently as to allow therapeutic intervention. Even if pathway inhibition is prohibited by normal physiological requirements, other mechanism-based approaches that exploit aberrant pathway activation might be feasible. It has been proposed that malignancy is determined in all tissues by mis-regulation of a common set of genes that control growth by affecting cell proliferation, apoptosis, invasion and angiogenesis. This hypothesis is supported by the demonstration that multiple types of normal human cells can be made tumorigenic by the expression of a defined set of viral and cellular proteins. Therapeutic agents for the treatment of such tumors might target not only self-renewal pathway components, but also other critical transcriptional targets of the self-renewal pathways, or proteins that co-operate with them to deregulate growth.It is important that agents directed against cancer stem cells discriminate between cancer stem cells and normal stem cells. This will require the identification of realistic drug targets unique to cancer stem cells. The identification of such targets and the development of anti-cancer agents will require a deeper understanding of normal stem cell biology as well as cancer biology. More importantly, identification of the ovarian cancer stem cell would provide a critical step in advancing the development of novel therapeutic strategies in the management of ovarian cancer. Furthermore, characterizations of such progenitor or cancer stem cells in drug resistant  manner for ovarian cancer will likely lead to a greater understanding of early events leading to the genesis of this elusive disease, in addition to providing new therapeutics targets aimed at the cells directly responsible for its propagation.CSC: Cancer Stem Cell; CNS: Central Nervous System; ESA: Epithelial-Specific Antigen; Shh: Sonic Hedgehog; Hh: Hedgehog; EOC: Epithelial ovarian Cancer; OCIC: Ovarian Cancer Initiating Cells; SP: Side PopulationThe authors declare that they have no competing interests.PPM participated in drafting the full manuscript and creating figures. SKB participated in substantial contribution to conception and revising it critically for important intellectual content.


The broad aim of this study was to assess the contribution of job strain to mental health inequalities by (a) estimating the proportion of depression attributable to job strain (low control and high demand jobs), (b) assessing variation in attributable risk by occupational skill level, and (c) comparing numbers of job strain–attributable depression cases to numbers of compensated 'mental stress' claims.Standard population attributable risk (PAR) methods were used to estimate the proportion of depression attributable to job strain. An adjusted Odds Ratio (OR) of 1.82 for job strain in relation to depression was obtained from a recently published meta-analysis and combined with exposure prevalence data from the Australian state of Victoria. Job strain exposure prevalence was determined from a 2003 population-based telephone survey of working Victorians  using validated measures of job control  and psychological demands . Estimates of absolute numbers of prevalent cases of depression and successful stress-related workers' compensation claims were obtained from publicly available Australian government sources.Overall job strain-population attributable risk (PAR) for depression was 13.2% for males  and 17.2%  for females. There was a clear gradient of increasing PAR with decreasing occupational skill level. Estimation of job strain–attributable cases  versus "mental stress" compensation claims (696) suggest that claims statistics underestimate job strain–attributable depression by roughly 30-fold.Job strain and associated depression risks represent a substantial, preventable, and inequitably distributed public health problem. The social patterning of job strain-attributable depression parallels the social patterning of mental illness, suggesting that job strain is an important contributor to mental health inequalities. The numbers of compensated 'mental stress' claims compared to job strain-attributable depression cases suggest that there is substantial under-recognition and under-compensation of job strain-attributable depression. Primary, secondary, and tertiary intervention efforts should be substantially expanded, with intervention priorities based on hazard and associated health outcome data as an essential complement to claims statistics. Data anJob strain prevalence was significantly higher among women than men (25.5% versus 18.6%), including after adjustment for occupational skill level  and corThe PAR estimates presented are conservative in several ways. The analyses are based on a conservative effect size estimate that is lower than those published for most individual studies. Some cross-sectional studies have found strong associations between job strain and prevalent cases of depression, such as a US study that presented adjusted ORs for job strain and major depressive episode (OR = 7.0), depressive episode (OR = 4.1), and dysphoria (OR = 2.9) among women, with no significant associations among men . Longituall psychosocial working conditions on depression would be higher than the estimates we have presented, and corresponding estimates for all affected mental health outcomes would be higher still.The PAR estimates presented are also conservative in that the steepness of the gradient observed may be underestimated. It is postulated that job stress has a greater impact on mental health among people in lower socioeconomic positions ; as has We also acknowledge certain limitations of this study. Although the survey was designed to be representative of the working population, the sample was taken from telephone listings. This may disproportionately exclude those workers who are in less secure employment, in lower status groups, shift workers and those working longer hours. This bias would lead to underestimation of differentials in exposure and disease distributions across occupational levels. Further, job strain has been studied in relation to various measures of depression, including the CES-D scale ,33, GoldWe also acknowledge the need for further study of job stress in general and job strain in particular in relation to mental health outcomes. Though the majority of studies of job strain or its component scales of job control and job demands show positive associations, studies of job strain and depression show inconsistent effect sizes, with some studies finding a null association. Substantial differences in the effect size estimates for the contributing studies to the meta-analysis used in the PAR calculations led to its wide confidence intervals , and in turn to wide confidence intervals in our PAR estimates. But we also argue from a precautionary principle perspective that the evidence presented is adequate to justify expanded policy and practice responses, particularly given the scale of the problem and the availability of intervention strategies to address it . SeveralFinally, comparison of job strain-attributable depression estimates to compensated "mental stress" claims shows that the scale of the problem is grossly underestimated by WC statistics. There are a number of contributing factors for this discrepancy. WC statistics are based on accepted claims. Information regarding the numbers of claims submitted is unavailable. However, given the adversarial nature of the WC system, it is likely that many workers with stress-related illnesses, including depression, have their claims rejected, or may be deterred from filing a claim. Further, when a worker presents to a medical practitioner with depression, the practitioner may or may not identify an underlying occupational causation or contribution. Even if job stress is medically recognized as a contributory factor, there is a documented reluctance amongst Australian general practitioners to initiate WC claims for patients presenting with job stress-related conditions ,48. FurtThis discrepancy between epidemiologic versus insurance-based estimates of job strain–attributable depression demonstrates an urgent need for population-based exposure and health data to complement claims statistics as a basis for guiding policy and practice strategies and priorities in this area. Further, the social patterning of job strain-attributable depression indicates that concerted action by occupational health & safety regulatory agencies and other workplace stakeholders, particularly in lower-skilled work contexts, has the potential to reduce mental health inequities on a population level. Applying a health inequalities intervention framework consistent with the systems approach described above, Whitehead recently described how stressful psychosocial working conditions can be addressed by a combination of strengthening individuals, strengthening communities, improving living and working conditions, and promoting healthy macro-policies . There iGiven the availability of feasible and effective intervention strategies, our findings add to the growing evidence base in support of expanded public health policy and practice interventions to address the adverse effects of job stress. A substantial and inequitable disease burden could be addressed by applying a systems approach to job stress in Victoria and elsewhere. In addition to being a concern for workers, unions, employers, occupational health and safety, and workers' compensation systems, job stress should be a concern for physical and mental health promotion agencies, government public health authorities , medical practitioners, community advocacy groups, and others. An optimal public health response to job stress would encompass participation by the full range of stakeholders.The authors declare that they have no competing interests.ADLaM conceived of the study, directed analyses, and led the writing of the paper. TK and DV assisted with the conceptualisation, analysis, and writing. AO and RW assisted with conceptualisation and analyses. All authors contributed to the interpretation of results, refinement of analyses, and review of successive drafts of the manuscript.The pre-publication history for this paper can be accessed here:


Recent evidence has demonstrated that endothelial-to-mesenchymal transition (EndMT) may have a significant role in a number of diseases. Although EndMT has been previously studied as a critical process in heart development, it is now clear that EndMT can also occur postnatally in various pathologic settings, including cancer and cardiac fibrosis. During EndMT, resident endothelial cells delaminate from an organised cell layer and acquire a mesenchymal phenotype characterised by loss of cell–cell junctions, loss of endothelial markers, gain of mesenchymal markers, and acquisition of invasive and migratory properties. Endothelial-to-mesenchymal transition -derived cells are believed to function as fibroblasts in damaged tissue, and may therefore have an important role in tissue remodelling and fibrosis. In tumours, EndMT is an important source of cancer-associated fibroblasts (CAFs), which are known to facilitate tumour progression in several ways. These new findings suggest that targeting EndMT may be a novel therapeutic strategy, which is broadly applicable not only to cancer but also to various other disease states. As an integral component of the circulatory system, the endothelium can be defined as the single-cell layer of mostly squamous epithelium that provides the inner cell lining of blood vessels and lymphatics . Endotheα-smooth muscle actin , and are a principle source of host-derived vascular endothelial growth factor (VEGF), which promotes angiogenesis . Cancer-associated fibroblasts play an important role in tumour progression and can alter the microenvironment in several ways. In particular, CAFs deposit various extracellular matrix molecules and secrete paracrine factors that can directly affect the behaviour of many different cell types within the tumour. Furthermore, CAFs release potentially oncogenic signals, such as transforming growth factor-ogenesis . Here, wEndothelial-to-mesenchymal transition is often categorised as a specialised form of epithelial-to-mesenchymal transition (EMT). Epithelial-to-mesenchymal transition can occur in many epithelial cell types and is a critical process in embryogenesis . In the Epithelial-to-mesenchymal transition has been extensively studied and has provided a useful framework for guiding research on EndMT. Both EMT and EndMT give rise to cells that have a similar mesenchymal phenotype, and current evidence suggests that both utilise common signalling pathways. However, further studies are needed to validate this notion, as there exist some key differences between endothelial cells and other types of epithelial cells. In particular, endothelial cells express distinct cell–cell junctional proteins, different cytoskeletal proteins, different signalling machinery, and different surface markers . The impβ, bone morphogenic protein (BMP), and Notch pathways  exhibited a significant reduction both in fibrosis as well as EndMT (Another recent study has further validated the notion that EndMT can occur postnatally in the context of disease. This study addressed EndMT during cardiac fibrosis, a common feature of most forms of heart failure ( in vivo . Interesas EndMT .In addition to the studies described above, there is evidence to suggest that EndMT may occur in many other disease settings, such as chronic pulmonary hypertension , and in in vivo studies described above, a number of reports have also demonstrated the induction of EndMT in vitro. To date, many different endothelial cell types, including both human and mouse, have demonstrated EndMT when exposed to TGF-β or Notch ligands in vitro (In vivo studies have further substantiated that EndMT can be modulated in response to manipulations of the TGF-β or Notch pathways (β, or a combination of both pathways provides the initiating signal under physiologic conditions in vivo. It is also likely that other signalling pathways interact with TGF-β and Notch to mediate EndMT. For example, VEGF, NFAT, BMP, Wnt/β-catenin, ErbB, and NF1/Ras have been implicated in EndMT during cardiac development (In addition to the in vitro . These selopment , but havSnail family of transcriptional repressors (Snail repressors are believed to downregulate VE-cadherin, thereby disrupting adherens junctions and allowing endothelial cells to delaminate and undergo EndMT. It is unknown whether other effectors of EMT, such as Twist (Downstream of these signalling events, the transcriptional networks mediating EndMT also remain largely unidentified. In one series of experiments, cells that undergo EndMT exhibited an increased expression of the pressors . In the as Twist , CArG boStudies of EndMT have revealed a novel mechanism of fibroblast and mural cell recruitment that is likely to be involved in many different disease settings. Fibroblasts are known to have an important role in tissue remodelling and fibrosis , althougβ and/or BMP signalling pathways. The mouse studies described above have demonstrated that systemic administration of rhBMP-7 significantly reduced EndMT during cardiac fibrosis (Perhaps most importantly, the recent discoveries of EndMT in different diseases suggest that modulating EndMT may represent a promising new treatment modality. The endothelium itself is an attractive target for drug delivery because it lies in direct contact with the bloodstream. We hypothesise that therapies directed at inhibiting EndMT may delay tumour progression, perhaps as a result of impaired angiogenesis or CAF recruitment. Possible treatment strategies may target the TGF-Furthermore, inhibiting EndMT may be broadly applicable to various disease states. For example, preventing EndMT during chronic organ fibrosis may significantly delay disease progression and allow patients to maintain adequate organ function for a longer period of time. Nevertheless, additional studies are needed to identify the precise molecular mechanisms of EndMT in disease and to determine which signalling components might be viable therapeutic targets. A promising place to start may be to further examine the developmental defects in EndMT. Just as normal developmental mechanisms are often recapitulated in certain disease states (


Consumer directed health care proposes that patients will engage as informed consumers of health care services by sharing in more of their medical costs, often through deductibles. We examined knowledge of deductible plan details among new enrollees, as well as anticipated care-seeking changes in response to the deductible.In a large integrated delivery system with a range of deductible-based health plans which varied in services included or exempted from deductible, we conducted a mixed-method, cross-sectional telephone interview study.Among 458 adults newly enrolled in a deductible plan (71% response rate), 51% knew they had a deductible, 26% knew the deductible amount, and 6% knew which medical services were included or exempted from their deductible. After adjusting for respondent characteristics, those with more deductible-applicable services and those with lower self-reported health status were significantly more likely to know they had a deductible. Among those who knew of their deductible, half anticipated that it would cause them to delay or avoid medical care, including avoiding doctor's office visits and medical tests, even services that they believed were medically necessary. Many expressed concern about their costs, anticipating the inability to afford care and expressing the desire to change plans.Early in their experience with a deductible, patients had limited awareness of the deductible and little knowledge of the details. Many who knew of the deductible reported that it would cause them to delay or avoid seeking care and were concerned about their healthcare costs. An increasing number of patients are facing higher health insurance deductibles . Often, For the incentives to work, it is important for patients to understand their cost-sharing plan and be able to anticipate how much they will have to pay for various services when they need them. Currently, little is known about how well patients understand their deductible plans, and how they adapt their care-seeking behavior under these plans ,9. SinceIn this study we examine deductible plan enrollees' knowledge of their plan at the beginning of the coverage year, and describe qualitative reports of anticipated changes in care-seeking in response to the deductible.Kaiser Permanente Northern California (KPNC) is an integrated delivery system (IDS) which newly began offering deductible health plans to enrollees of large employer groups in 2005. Cost-sharing plans varied by employer, but enrollees could not self-select their KPNC cost-sharing level. All study respondents had one of two types of deductible plans: the more generous plan had a deductible for hospital care and emergency department (ED) visits only; the less generous plan additionally included medical tests in the deductible. In both plans, patients paid a copayment for all office visits and generic drugs; some also faced a separate drug deductible for brand-name drugs only. Family plan deductible amounts were twice the individual deductible amount.Our source population included all adult (age ≥18) health plan members enrolled in a deductible plan through a large-group employer in January 2005 who had also been members in January 2004. We obtained a stratified random sample of 1,100 members: 550 from the population without a chronic disease and 550 from patients in the health plan's chronic disease registries for asthma, hypertension or diabetes. Since we did not find a statistically significant difference between the chronic and non-chronic disease populations in any of the analyses for this paper, we pooled these two groups. Telephone study eligibility criteria excluded those who did not have valid contact information (n = 139), were unreachable (n = 187), or with language barriers, cognitive impairment, or severe illness (n = 47); we also excluded those who died (n = 1) or left the health system or deductible plan (n = 72).In early 2005, we mailed a study introduction letter with a pre-paid reply postcard to our study sample. Research assistants called all who did not refuse participation (up to 15 times during various times of the day and week) obtaining verbal consent before beginning the interview. The Kaiser Foundation Research Institute Institutional Review Board approved the study protocol. Overall, 71% (n = 458) of all eligible subjects participated in the study: 74% from the chronic disease sample and 67% from the non-chronic disease population. Respondents were similar to non-respondents with respect to age, gender, plan characteristics, and chronic disease status (p > 0.05).To assess deductible knowledge, we asked if respondents had any medical deductible, the deductible amount, and if they had to pay the full price before reaching their deductible for hospital care, ED visits, medical tests, preventive office visits or non-preventive office visits. If respondents were unsure, the interviewer encouraged them to make an educated guess. For respondents who reported a deductible, we asked an open-ended question about how it would affect their care-seeking .We also collected respondents' race/ethnicity, education level, marital status, annual household income, and health status. We obtained information on respondents' age, gender, and actual cost-sharing from health system administrative databases.To assess cost-sharing knowledge, we calculated the percent of respondents who correctly reported having a deductible, and which services were included in the deductible. We considered the respondent correct if their self-reported deductible matched either their individual or family deductible amount. Since the study over-sampled patients with chronic disease, all analyses were either stratified or weighted. All multivariate analyses also adjust for this stratification variable. To examine the association between deductible knowledge and respondent characteristics, we used multiple logistic regression. We included the missing responses as a separate category for each variable.We coded respondents' open-ended descriptions of the anticipated effect of their deductible on care-seeking into 13 specific categories, with responses coded into as many categories as applied (from one to four categories per respondent). We then condensed these categories into four general themes . Of all respondents, 26% did know their deductible amount and 6% knew which specific services were included or excluded from the deductible. Altogether, 5% of all respondents correctly knew that they had a deductible, knew the deductible amount and knew which services applied to the deductible. Figure While hospital care and emergency department visits were included in all respondents' deductibles, 81% did not know that they had to pay the full price before reaching the deductible. Among patients' whose deductible included medical tests (less generous plan), 72% did not know that they had to pay the full price; among those whose deductible excluded medical tests (more generous plan), 38% mistakenly thought that they had to pay the full price for medical tests. Office visits were excluded from all deductibles in this population, but 11% of all respondents mistakenly thought that they had to pay the full price for office visits.In multivariate analyses . Table Overall, 51% of respondents reported that the deductible would cause them to decrease or delay their use of medical care, including going to the doctor less, getting fewer medical tests, going to the emergency room less, and thinking twice about or delaying care. Often these respondents described that they would reduce care-seeking even for types of care that they believed were clinically necessary  knew exactly which services were included in or exempt from their deductible, which raises concerns about patients' ability to successfully navigate the health-care system while facing the complex benefit structures of most deductible plans. For example, patients who are unaware of plan exemptions may unnecessarily avoid preventive care. Our previous studies showed similarly that patients have limited knowledge of their cost-sharing details in non-deductible plans ,9. EmploAlmost half of respondents reported that the deductible would cause them to decrease their use of health care. One in ten reported the deductible would cause them to reduce physician visits, even though office visits were excluded from the deductible. Over 25 percent reported having concerns about costs under the deductible. Still, almost one out of four respondents did report that deductible costs would not affect their care-seeking behavior. Despite frequent reports of planned changes in care-seeking, patients' actual care-seeking decisions may differ. It is important to continue monitoring the ongoing effects of deductibles and other consumer directed health plans on patient care-seeking behavior to understand which types of patients change their behavior, and whether patients respond by reducing discretionary services or avoiding clinically urgent medical care.While some early studies of deductible plan configurations did not find that patients were deterred form seeking preventive-care ,12, patiPatients newly enrolled in a deductible plan had limited knowledge of having any deductible, including rarely being aware of which services are deductible-exempt. The limited knowledge raises concerns about patients' ability to engage as informed consumers of healthcare, especially in an environment of consumer-directed health plan designs with additional complex features such as healthcare savings accounts. At minimum, more effective efforts to educate consumers about the working of their plan are needed.Half the patients who knew of their deductible reported that they planned to delay or avoid care. It is important to continue to monitor patient perceptions of their benefit plan and to determine if poor knowledge of deductible health plans combined with decreased health care utilization adversely affects health care outcomes.Newhouse is a director of and holds equity in Aetna, which sells high deductible health plans. The other authors declare that they have no competing interestsMR made substantial contributions to the study's conception and design; data collection, analysis and interpretation; drafted and revised the manuscript critically for important intellectual content. NB made substantial contributions to the analysis and interpretation of the data, and revised the manuscript critically for important intellectual content. RB made substantial contributions to the study's design, analysis and interpretation of the data, and revised the manuscript critically for important intellectual content. JPN made substantial contributions to the study's design, analysis and interpretation of the data, and revised the manuscript critically for important intellectual content. JH made substantial contributions to the study's conception and design; data collection, analysis and interpretation; and revised the manuscript critically for important intellectual content. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6963/9/244/prepubRespondent characteristics associated with knowledge of having a medical deductible. Table displays the adjusted odds ratio for correctly reporting having a medical deductible or prescription drug deductible from multiple logistic regression weighted for sampling proportions.Click here for file


During its earliest stage, before cell loss and independent of amyloid plaques and neurofibrillary tangles, Alzheimer's disease (AD) causes synaptic loss affecting the basal functional properties of neurons. In principle, synaptic loss can be detected by measuring AD-induced changes in basal function, or by measuring stimulus-evoked responses on top of basal changes. Functional magnetic resonance imaging (fMRI) is sensitive to both basal changes and evoked-responses, and there are therefore two experimental approaches in which fMRI can be used to pinpoint synaptic loss in AD. In this review, we will compare and contrast both approaches for pinpointing when and where synaptic loss in AD begins and for monitoring therapeutic efficacy.


BMC Developmental Biology reports that a mitochondrial iron importer is required for Drosophila male fertility and normal mitochondrial shaping in spermatids. This suggests that mitochondrial morphogenesis during insect spermatogenesis may be a useful new context in which to study iron metabolism.A recent paper in http://www.biomedcentral.com/1471-213X/10/68See research article  Mechanisms of iron homeostasis have been characterized primarily in vertebrate blood, liver, and intestinal tissue, as well as in budding yeast. In vertebrates, iron transport and trafficking in and out of cells requires such proteins as transferrin, transferrin receptors, ferroportin, hepcidin and ferritin . Once inBMC Developmental Biology, Metzendorf and Lind [Drosophila spermatogenesis as an emerging context for the study of mitochondrial iron metabolism through their analysis of the Drosophila gene mitoferrin (dmfrn). During spermatid development in flies, mitochondria undergo dramatic events of movement and shaping, including aggregation, fusion and elongation beside the growing flagellar axoneme. Disruptions of mitochondrial function or dynamics typically affect male fertility, a phenotype that is easily screened for and characterized at the subcellular level. The dmfrn gene product and other iron metabolism proteins show enriched expression in the testes of mammals and insects, suggesting special roles for mitochondrial iron metabolism in spermatogenesis. Analysis of these requirements in insect spermatogenesis could lead to new insights into human genetic disorders such as Friedreich's ataxia and sideroblastic anemia, both associated with defects in iron homeostasis within mitochondrial compartments.In a recent study published in and Lind  establisD. melanogaster mitoferrin (dmfrn) mutants and showed that the Dmfrn mitochondrial iron importer is required for spermatid mitochondrial morphogenesis and thus for development of mature motile sperm [Drosophila genome includes a single homolog, dmfrn, of two human paralogs, mitoferrin-1 and mitoferrin-2, and two Saccharomyces cerevisiae paralogs, mrs3 and mrs4; all tests of cross-species complementation so far have indicated strong functional conservation. Studies in vertebrates and yeast demonstrate a role for mitoferrin family members in import of iron from the cytosol into mitochondria [dmfrn. All alleles were associated with male sterility, and the deficiency alleles were associated with only a mild decrease in viability. The testis phenotype was enhanced and suppressed, respectively, by low and high iron levels in food.Metzendorf and Lind characterized le sperm . As in odmfrn testes was variable in severity, with some flies showing more robust testes and more frequent elongated cysts of spermatids than others. Failure of mitochondrial elongation accompanying axoneme elongation was common, as was failure of spermatid cyst individualization. In wild-type early round spermatids, the large, layered, spherical mitochondrial structure called the Nebenkern disentangles and elongates beside the nascent spermatid flagellar axoneme. In dmfrn mutants, the Nebenkern often remained aberrantly round while the axoneme microtubules grew , l Figure . This padmfrn mutants. Individualization is the process at the end of spermatogenesis during which the 64 interconnected spermatids in a cyst become surrounded by their own plasma membranes. Failure of individualization is a typical secondary phenotype in mutant testes with an earlier structural defect.Additional TEM data, combined with staining of actin and other structures, indicated that sperm individualization typically failed in mitoferrin showed enriched expression in testis tissue. A dmfrn-lacZ reporter construct indicated protein expression in post-meiotic spermatids. A tagged and functional dmfrn was associated with mitochondria in early round spermatids and later stages, with much of the protein sent to the waste bag during individualization. At the RNA level, the authors found similar testis enrichment of two additional iron-metabolism genes, fh and Fer3HCH. Expression patterns of these genes plus another, Abcb10, together suggest that Drosophila spermatogenesis could be a fruitful context for further analysis of iron metabolism.The lack of dramatic phenotypes in tissues other than the testis led the authors to confirm by RT-PCR that Drosophila fh gene is the ortholog of S. cerevisiae YFH1 and of human FXN, a gene associated with the neurodegenerative disorder Friedreich's ataxia. FXN encodes frataxin, a mitochondrial matrix protein whose role appears to be the delivery of iron to the scaffolding proteins on which Fe-S clusters are formed [Drosophila frataxin is expressed in multiple tissues besides the testis and, unlike mitoferrin, appears essential for normal viability. Studies of Drosophila frataxin are now possible through RNA interference [The e formed . These Frference ; perhapsDrosophila is evident through sequence homology searches of the Drosophila genome [Drosophila [Abcb10 may enable exploration of genetic interactions with dmfrn and the physical interactions of their gene products. Ferritin is an iron-sequestration protein that has been extensively studied in its cytosolic and secreted forms. Recent years have seen the discovery of a new ferritin in mammals and insects that is targeted to mitochondria. Drosophila mitochondrial ferritin, encoded by Fer3HCH, is highly expressed in the testis, and the gene product localizes to mitochondria when transfected into human cells [mitoferrin, fh, and Abcb10. Mammalian mitochondrial ferritin is similarly expressed highly in testes [In mammals, the ABC-family transporter protein Abcb10 forms a complex with mitoferrin-1 and increases iron import into mitochondria, as recently characterized in erythroid tissue . A closea genome ; the Flyosophila . Analysian cells . This geDrosophila sperm development, and their confirmation of testis expression of other proteins involved in iron homeostasis, set the stage for future exploration of iron metabolism as an important factor in mitochondrial morphogenesis and fertility in both vertebrates and invertebrates.Metzendorf and Lind's identification of a role for the mitochondrial iron importer dmfrn in


By studying genome-wide expression patterns in healthy and diseased tissues across a wide range of pathophysiological conditions, DNA microarrays have revealed unique insights into complex diseases. However, the high-dimensionality of microarray data makes interpretation of heterogeneous gene expression studies inherently difficult.Using a large-scale analysis of more than 40 microarray studies encompassing ~2400 mammalian tissue samples, we identified a common theme across heterogeneous microarray studies evident by a robust genome-wide inverse regulation of metabolic and cell signaling pathways: We found that upregulation of cell signaling pathways was invariably accompanied by downregulation of cell metabolic transcriptional activity (and vice versa). Several findings suggest that this characteristic gene expression pattern represents a new principle of mammalian transcriptional regulation. First, this coordinated transcriptional pattern occurred in a wide variety of physiological and pathophysiological conditions and was identified across all 20 human and animal tissue types examined. Second, the differences in metabolic gene expression predicted the magnitude of differences for signaling and all other pathways, i.e. tissue samples with similar expression levels of metabolic transcripts did not show any differences in gene expression for all other pathways. Third, this transcriptional pattern predicted a profound effect on the proteome, evident by differences in structure, stability and post-translational modifications of proteins belonging to signaling and metabolic pathways, respectively.Our data suggest that in a wide range of physiological and pathophysiological conditions, gene expression changes exhibit a recurring pattern along a transcriptional axis, characterized by an inverse regulation of major metabolic and cell signaling pathways. Given its widespread occurrence and its predicted effects on protein structure, protein stability and post-translational modifications, we propose a new principle for transcriptional regulation in mammalian biology. Transcriptional profiling by DNA microarrays allows the simultaneous quantitative analysis of tens of thousands of transcripts in a single experiment. By applying transcriptional profiling technology to healthy and diseased tissues across a wide range of pathophysiological conditions, DNA microarrays have revealed unique insights into complex disease patterns. However, the high-dimensionality of microarray data makes interpretation of heterogeneous gene expression studies inherently difficult. One of the main challenges in the analysis of microarray data is to identify common underlying biological themes by integrating multiple similar experiments. A frequent approach to this problem is to extract common genes from these gene lists and then subject these genes to enrichment analysis by grouping them into pathways.Kyoto Encyclopedia of Genes and Genomes [In a previous study examining failing and non-diseased dog hearts, we observed an intriguing reciprocal transcriptional regulation of selected cell signaling and metabolic processes . To exte Genomes ) in a laWe examined gene expression patterns across 20 large microarray datasets of different human tissues by comparing, in each tissue type, the 10 samples with the highest vs. the lowest gene expression of transcripts belonging to the KEGG pathway of oxidative phosphorylation (OXPHOS) using Significance Analysis of Microarrays . The difall human and animal microarray studies, defined as the number of up- minus down-regulated genes of these KEGG pathways expressed as percentage of the total number of regulated genes within a study, a negative correlation was found  and 212 kinase phosphorylation sites and found that these post-translational modification sites were significantly enriched in signaling compared to metabolic pathways  lack a rigid 3D structure and possess an increased exposed surface area, facilitating interaction with multiple targets ,5. These Figures . Of note Figures . OverallCells react to changes in their environment by a coordinated transcriptional response. Using a meta-analysis of more than 40 diverse microarray studies which included different microarray platforms  and different methods of normalizations , we demonstrate a robust interaction between gene expression in signaling and metabolic pathways. While metabolic pathways were positively correlated to each other, they were negatively correlated to signal transduction pathways. Several findings suggest that this characteristic gene expression pattern represents a novel paradigm for mammalian transcriptional regulation. First, this coordinated transcriptional pattern occurred in a wide variety of physiological and pathophysiological conditions and was identified in all 20 different tissue types examined. Importantly, it occurred independently of the proliferative potential of the underlying tissue, as the inverse regulation of metabolism and signal transduction was observed in terminally differentiated organs like brain and heart, but also in more rapidly dividing malignant tumors. Second, and most strikingly, these changes in steady-state mRNA levels predict a profound effect on the proteome, as KEGG cell signaling pathways are characterized by an increased magnitude of IUPs as compared to metabolic and biosynthetic pathways. The lack of a rigid 3D structure in IUPs is thought to provide several functional advantages, including conformational flexibility to interact with multiple targets, increased interaction surface area, and accessible post-translational modification sites . An incrIn summary, proteins in signaling and metabolic pathways have fundamentally different properties ranging from inversely regulated transcriptional patterns Figures  and 3, aIn the present study, we noted a striking and robust reciprocal correlation of transcriptional changes between metabolic and signaling pathways. Importantly, correlations do not prove cause and effect. Therefore, we can not determine whether transcriptional changes in metabolic activity anticipate changes in signaling pathways or vice versa. While this study was centered on pathway analysis, future studies will need to identify individual genes or hub nodes that connect metabolic and signaling pathways. In addition, the role of up- and down-stream regulatory events, e.g. transcription factors, miRNAs, splicing, 3' end termination and/or stability of mRNAs need to be examined.Future studies will need to address the role of this transcriptional pattern in various disease processes. While the association of IUPs with various disease processes might suggest that down-regulation of metabolism and up-regulation of signaling pathways is a common theme in a wide range of disease processes, we found this generalization is not universal. This could be related to a different baseline level of OXPHOS activity in various tissues and cancer specimens and/or differences in tissue handling. Clearly, future studies need to address whether this transcriptional pattern will help in refining the distinction between diseased and non-diseased tissue samples.Public datasets were obtained from the GEO database . A detaiminus down-regulated transcripts of a KEGG pathway expressed as percentage of the total number of regulated genes within a study. Clustering of the expression of KEGG pathways and phosphorylation sites was done using Genesis [To determine differentially expressed genes, unpaired two-class Significance Analysis of Microarrays (SAM) was used . Differe Genesis .http://www.cbs.dtu.dk/services/NetNGlyc, NetOGlyc 3.1 [Batch prediction of long disordered regions was carried out using the IUPforest-L software, based on the Moreau-Broto autocorrelation function of amino acid indices (AAIs) and other physicochemical features of the primary sequences . Non-parGlyc 3.1 , SUMOsp Glyc 3.1 , and GPSGlyc 3.1 , respectIUP: Intrinsically Unstructured Proteins; KEGG: Kyoto Encyclopedia of Genes and Genomes; OXPHOS: Oxidative Phosphorylation; SAM: Significance Analysis of Microarrays; DAVID: Database for Annotation, Visualization and Integrated Discovery; GEO: Gene Expression Omnibus.ASB conceived the study, carried out the experiments and drafted the manuscript. AK and CC provided assistance with the bioinformatic and statistical analysis, respectively; KBM and TPC participated in study design. GFT conceived the study and drafted the manuscript. All authors read and approved the final manuscript.Graphical representation of 200 KEGG pathways sorted based on their similarity to OXPHOS expression. For 20 different human tissues, KEGG pathways were compared between the ten samples displaying the highest and the lowest values of OXPHOS gene expression  was color-coded with yellow and blue representing low and high expression of the pathways, respectively. KEGG pathways were then sorted according to their similarity to "oxidative phosphorylation" which is represented by the top row in Additional File Click here for fileList of gene expression datasets used in the present study. The study-ID, tissue type, Gene Expression Omnibus (GEO) accession number, species, sample characteristics, comparison, microarray type and methods of normalization are given for each dataset.Click here for fileList of human tissues samples with high vs. low OXPHOS gene activity. The tissue type, study-ID (Gene Expression Omnibus (GEO) accession number), sample-ID and clinical characteristics are given for samples with high and low OXPHOS gene activity.Click here for file


Among bacteria and archaea, amino acid usage is correlated with habitat temperatures. In particular, protein surfaces in species thriving at higher temperatures appear to be enriched in amino acids that stabilize protein structure and depleted in amino acids that decrease thermostability. Does this observation reflect a causal relationship, or could the apparent trend be caused by phylogenetic relatedness among sampled organisms living at different temperatures? And do proteins from endothermic and exothermic vertebrates show similar differences?We find that the observed correlations between the frequencies of individual amino acids and prokaryotic habitat temperature are strongly influenced by evolutionary relatedness between the species analysed; however, a proteome-wide bias towards increased thermostability remains after controlling for phylogeny. Do eukaryotes show similar effects of thermal adaptation? A small shift of amino acid usage in the expected direction is observed in endothermic ('warm-blooded') mammals and chicken compared to ectothermic ('cold-blooded') vertebrates with lower body temperatures; this shift is not simply explained by nucleotide usage biases.Protein homologs operating at different temperatures have different amino acid composition, both in prokaryotes and in vertebrates. Thus, during the transition from ectothermic to endothermic life styles, the ancestors of mammals and of birds may have experienced weak genome-wide positive selection to increase the thermostability of their proteins. Evolutionary molecular biology is mostly concerned with the forces affecting individual genes. However, observations of variable proportions of guanine and cytosine (GC) in different species and in different genomic regions of vertebrates  have A clear picture of selection at work emerges only in the study of structured RNAs. The ribosomal RNAs and transfer RNAs of prokaryotes living at high temperatures contain a much larger GC-fraction in their stem regions compared to homologs from prokaryotes living at more moderate temperatures ,8, likelHowever, a higher GC-content in structural RNAs of thermophiles and hyperthermophiles may also have arisen through reasons unrelated to environmental temperatures, e.g., random genetic drift or mutational biases. Closely related species often have similar nucleotide composition and similar habitats simply due to their descent from a common ancestor; a statistically significant relationship between GC content and temperature across species might thus reflect nothing more than a close phylogenetic relationship of these species. This is not the case: even after controlling for phylogenetic relationships, the GC content of structural RNA remains strongly correlated with optimal growth temperature . Thus, gJust like structural RNAs, proteins need to retain their three-dimensional structure in the presence of thermal fluctuations. It hence appears likely that the proteins of thermophilic organisms show corresponding signs of thermal adaptation. Several studies indeed report a correlation between amino acid usage and optimal growth temperature of bacteria -14; howeet al. . As biaDanio rerio orthologs to those codons with the same purine content, we still observe a significantly higher ERK value in the human sequences (Wilcoxon rank sum test: p < 10-15). Thus, the biased amino acid composition of proteins from endothermic vertebrates cannot be attributed to purine loading alone.To exclude purine loading as a confounding factor, we employed an analogous strategy as for GC content. When we restrict the alignments of the 6227 human - ERK or CvP) correlates strongly with the optimal growth temperature of bacteria. That ERK and CvP measures are derived directly from physicochemical considerations [ERK and CvP are robust.Building upon earlier results on aligned structures of prokaryotic protein pairs ,16, we serations ,16 strenERK and CvP-bias compared to ectothermic fish, amphibia and reptilia. This increase cannot simply be explained by biases in nucleotide composition, and remains statistically significant when controlling for phylogenetic non-independence. While the examined dataset of genome sequences is necessarily small and not evenly sampled across vertebrates, we thus have strong evidence for a direct relationship between amino acid bias and the temperature at which vertebrate proteins operate. Analogous to the situation in prokaryotes, our findings are most parsimoniously explained by selection for increased stability against thermal fluctuations in endothermic vertebrates. Why then do we not see a correlation of amino acid usage bias with environmental temperature when considering only ectothermic vertebrates ? Apart from an issue of small sample size, this lack of a correlation may be due to the fact that ectothermic vertebrates can rapidly switch between habitats of different temperatures during evolution. This is evident, e.g., from the two Xenopus species in our study, which thrive at 18-22 and 23-28°C, respectively . For these 15 species, we performed an all-against-all protein blast search, identifying pair-wise orthologs through reciprocal best blast hits. We then excluded all proteins that had no orthologs outside their group . All remaining proteins had at least one ortholog outside their life style group, and where hence retained for our comparison , we constructed 508 protein segment alignments between Alligator mississippiensis and chicken, 429 segment alignments between Chrysemys picta (a turtle) and chicken, and 138 segment alignments between Anolis smaragdinus (another lizard) and chicken. As before, ERK and CvP-bias were calculated for each aligned segment.Based on well-aligning best blast hits of protein segments against the chicken genome (protein blast e-value < 10ERK values in endothermic compared to ectothermic vertebrates, we performed a detailed analysis of orthologs between human and Danio rerio. Orthologs were identified from reciprocal best blast hits. We aligned the translated orthologous protein sequences using MUSCLE [To check if nucleotide content variation could explain the higher g MUSCLE  with defERK from only those aligned codons that had the same G+C content in both species. Similarly, to exclude the influence of differences in purine content, we re-calculated ERK from only those codons that had the same A+G content in both species.To exclude the influence of differences in GC content between human and Danio on amino acid usage, we then re-calculated GW conceived of the study and performed the analyses. GW and MJL designed the study, interpreted the results, and drafted the manuscript. Both authors have read and approved the final manuscript.Supplemental Figures and Tables. Additional results that support the conclusions of the main text, presented in the form of figures and tables.Click here for file


Sir,We thank the correspondent for his interest in our report. Dr Tirelli has raised important questions regarding the specific subgroup of urological cancer patients included in our previous reported series (Dr Tirelli made the point that 66% of the castration-resistant prostate cancer (CRPC) patients included in the analysis were chemotherapy-naive. In 2004, two phase-III studies . Furthermore, many patients received abiraterone acetate, a novel CYP17-specific inhibitor, within this analysis or later; this agent has already shown preliminary promising activity  and the results from such analysis have to be treated with extreme caution. Prospective, randomised trials would be necessary to address the question of the optimal timing of docetaxel administration, and therefore the best timing for phase-I consideration relative to conventional chemotherapy in CRPC.Nonetheless, overall, these retrospective data provide preliminary clinical and ethical support for the exploration of novel agents both before and after docetaxel within early clinical trials, especially in CRPC patients where advances over the last two decades have been small.


Gonadotrophins, oestradiol, androstenedione, testosterone and dehydroepiandrosterone sulphate (DHAS) were measured sequentially in 72 patients with advanced breast cancer receiving endocrine therapy of various types. Tamoxifen significantly reduced gonadotrophins but did not effect other hormones. Danazol also reduced gonadotrophins. Aminoglutethimide (AGT) reduced oestradiol and DHAS but had not effect on gonadotrophins. The effects of administering tamoxifen, AGT and danazol together (TAD) together were therefore examined. This combination reduced gonadotrophins, oestradiol and DHAS, but no further than tamoxifen and AGT alone. The degree and duration of hormone suppression were similar in both responders and non-responders to tamoxifen, AGT or TAD, though patients responding to AGT showed more complete suppression at the end of the course of treatment, perhaps because they were treated longer. On relapse, adequate gonadotrophin and steroid suppression was demonstrated in patients receiving tamoxifen and AGT respectively. We conclude that (a) response to endocrine therapy is unlikely to be related to the degree of endocrine suppression produced by the therapy; (b) combination endocrine therapy does not further reduce serum-hormone concentrations and (c) relapse is unlikely to be due to escape from the hormone-inhibitory effects of endocrine agents.


A growing body of health and policy research suggests residential neighborhood density and street connectivity affect walking and total physical activity, both of which are important risk factors for obesity and related chronic diseases. The authors report results from their methodologically novel Twin Cities Walking Study; a multilevel study which examined the relationship between built environments, walking behavior and total physical activity. In order to maximize neighborhood-level variation while maintaining the exchangeability of resident-subjects, investigators sampled 716 adult persons nested in 36 randomly selected neighborhoods across four strata defined on density and street-connectivity – a matched sampling design. Outcome measures include two types of self-reported walking (from surveys and diaries) and so-called objective 7-day accelerometry measures. While crude differences are evident across all outcomes, adjusted effects show increased odds of travel walking in higher-density areas and increased odds of leisure walking in low-connectivity areas, but neither density nor street connectivity are meaningfully related to overall mean miles walked per day or increased total physical activity. Contrary to prior research, the authors conclude that the effects of density and block size on total walking and physical activity are modest to non-existent, if not contrapositive to hypotheses. Divergent findings are attributed to this study's sampling design, which tends to mitigate residual confounding by socioeconomic status. While both energy intake  and energy expenditure  are implicated in the current obesity epidemic, US national surveillance data regarding changes in individual-level energy intake and expenditure do not appear sufficient to explain or prevent obesity . AccordiThere is a strong conceptual and practical case for investigations into walking and "active transportation", and contexts in which they occur ,3. DespiA decrease in walking for transportation may relate to an increase in the proportion of the population residing in the suburbs as well as decreasing population densities in a large number of US metropolitan areas . Census New Urbanists, proponents of transit oriented development, and others interested in sustainability and smart growth, suggest that it is time to re-develop built environments that discourage active transport and carefully develop new areas to allow them to be more pedestrian friendly [It is hypothesized that environmental design makes a difference for physical activity, particularly when it comes to integrating walking into daily life -11. Whilfriendly . They hyfriendly .What are the key built environment factors thought to affect walking behavior? Research in transportation and physical activity has identified: (1) density, (2) street pattern or connectivity, (3) mixed land uses or the presence of destinations, and (4) pedestrian infrastructure and design related to the issues of comfort, safety, and interest. Of these, density and street pattern are considered very important. Density is thought to be important because higher densities tend to create a critical mass of people – more people to walk, to see others walking, to feel safer. Traffic congestion also increases with population and employment density so that at a certain threshold it is more convenient to walk  For examAt issue is the appropriate analytic methods for identifying the hypothesized multilevel effects in observational designs. In the past 6–7 years there has been a flurry of research that relies on regression adjustment, vis-à-vis the multilevel model. Oakes criticized this approach to this particular problem due to its unsupported interpolation thru design-space  -21. AlteThis paper reports the principal outcomes from the Twin Cities Walking Study (TCWS), an investigation that relies on matched sampling to test basic hypotheses about the relationship between density, street connectivity, and walking behavior.The TCWS is a cross-sectional observational study specifically designed to examine the influences of the built environment on walking and physical activity. All research activities were reviewed and approved by the University of Minnesota's IRB.Sampling is especially important in observational studies on the effects of the built environment. Researchers must appreciate that "like people" do not reside in diverse neighborhoods: the rich reside in one type of neighborhood, the poor another; whites in one, blacks in another; urbanites in one, farmers in another. In fact, it is such social stratification that defines neighborhoods in the first place -24. It fTCWS residential areas were selected from the environmentally diverse but demographically homogenous northern sector  of the Minneapolis-St. Paul metropolitan area, stretching from the urban core to the urban edge, for which especially rich geographic information system (GIS) data are available . Mean miles walked per day was calculated from 7-day travel and walking diaries each participant completed. Our diaries were modified versions of the National Household Travel Survey travel diary. Finally, total physical activity was measured by accelerometry. Total activity is among the most important outcome measures since it is related to many health risks and outcomes [The outcome measures for this analysis are limited to (1) travel walking, (2) leisure walking, (3) mean miles walked per day, and (4) total physical activity per day. Travel walking and leisure walking are self-report measures captured by the psychometrically established International Physical Activity Questionnaire – Long Form (IPAQ-LF) . The meaoutcomes . Particioutcomes  as well outcomes , when wooutcomes . Yet accFollowing best-practice insights, our accelerometer data were processed such that a "valid day" was any day with more than two hours of recorded movement above an analyst defined threshold of 3 activity counts: mean total activity counts per 24-hour day were calculated by summing counts within all valid days and then dividing by the number of valid days . Alternaet al. [Two environmental-level measures are central to this paper: density and street connectivity. Though there is debate regarding the best way to operationalize the construct, density is simply a number of items per unit land area. In this study, density is operationalized as total persons in housing units, as measured in the 2000 US Census at the block level per unit land area. Since the advent of widespread use of GIS in the 1990s, street pattern has been increasingly used as a measure of walkability. Recent work by Dill  and Steiet al.  list numet al. .We collected a vast array of subject- and area-level measures for use as covariates and secondary analyses. Included are age, self-reported overall health, race/ethnicity, educational attainment, and household income. Body mass index (BMI) was objectively measured during an in-person interview.Analytic results include descriptive statistics and odds-ratios (OR) from ordinal logistic regression models, sometimes called proportional odds models . OrdinalBasic information about our sample is presented in Table Table Table a priori defined regression models are presented in Table opposite direction to that proposed by new urbanists and others who believe dense highly-connected built environments promote physical activity. Overall, Table Adjusted odds ratios from Our study was not powered for subgroup analysis but results (not shown) seem hypothesis generating. Regression models akin to those described above show high density areas are marginally associated with an increase in total walking and, in some cases, total physical activity for racial minorities, those without a college degree, the less healthy (by self-report), and the obese. In short, members of these subgroups appear to benefit from high-density areas, at least with respect to walking and physical activity.A growing body of research suggests that various aspects of the built environment effect walking, physical activity, and health, especially BMI ,11,13,14This paper reports results of an observational study specifically designed to test associations between residential density, street-connectivity and walking behavior as measured through self-report and "objective" accelerometry. Although unadjusted differences in travel, leisure and total walking, and total physical activity, are evident, regression adjusted effects suggest dense areas promote travel walking while large-block  areas promote leisure walking. But the two effects appear to counterbalance one another such that total walking and total physical activity is not affected, as theory would suggest.Why do our results contradict results of even the most recent previous studies ,11,41? PSelection bias and other issues related to SES have clouded research in the subdiscipline. A few haThe author(s) declare that they have no competing interests.


Sir,Transfusion-related acute lung injury (TRALI) is an uncommon, probably underrecognized complication of transfusion of plasma-containing blood components, which is characterized by acute respiratory distress. It is a clinical diagnosis and may be diagnosed when acute lung injury (ALI) occurs during or within six hours of transfusion in a patient without pre-existing ALI, in the absence of temporally related alternative risk factors for ALI . This com3 with normal blood biochemistry. After receiving approximately 200 ml of blood for over two hours, she complained of breathlessness, which progressively increased over the next 20 minutes. On examination she was febrile, tachypneic (respiratory rate 32/minute), slightly cyanotic, with a pulse rate of 120/minute, blood pressure 90/60 mmHg, and SpO2 68%. The jugular venous pressure (JVP) was normal and chest auscultation revealed bilateral basal fine inspiratory crackles with normal breath sounds. Sinus tachycardia was noted on an electrocardiogram (ECG). An urgent X-ray chest showed bilateral patchy infiltrates in the mid- and lower zones with normal heart size, suggestive of noncardiogenic pulmonary edema. Echocardiography also ruled out cardiogenic dysfunction. Hematological investigations at this stage showed a total leukocyte count of 3200/mm3, with no evidence of any hemolytic process. In the absence of other mechanisms to explain ALI, a diagnosis of TRALI was made.A 52-year-old female patient with a diagnosis of inoperable carcinoma of the uterine cervix, received whole blood transfusion, in view of severe anemia. Before blood transfusion, she was alert, oriented, with no cardiorespiratory abnormality on clinicoradiological evaluation. Her vital parameters were normal and total leukocyte count was 8700/mmThe blood transfusion was stopped immediately and she was managed with oxygen therapy and intravenous fluids that resulted in a gradual improvement of dyspnea and oxygen saturation. She became completely chest asymptomatic after three days, with a normal chest X-ray.The reported incidence of TRALI in medical literature is one in 5000 transfusions and this may actually be much higher as TRALI is usually an underrecognized and underreported entity. TRALI haA mild form of TRALI can present with dyspnea and fever, while more severe forms can have severe respiratory distress that can quickly progress to respiratory failure. Other features include hypotension, frothy endotracheal secretions, and so on. Transient leucopenia, as noted in our patient, is occasionally observed and is believed to be due to the sequestration of leukocytes in pulmonary circulation. It is important to distinguish TRALI from volume overload, as the treatment of the two conditions is markedly different. Diuretics are contraindicated in TRALI and there is no role for steroids also. Severe forms of TRALI may require mechanical ventilation.Suspected cases of TRALI should be reported to the blood bank, to permit evaluation and deferral of high-risk donors, especially multiparous females. TRALI can be best prevented by avoiding unnecessary blood-product transfusions in selected populations.