We as dentists and surgeons would like to bring to the notice of our medical colleagues periodontal disease as yet another co-risk factor of pancreatic cancer.In a recent editorial by Dr Gaëtan Romain Joliat entitled ‘Latest Advances and Future Challenges in Pancreatic Surgery’, the authors stress on how development and improvement in the fields of chemotherapy, targeted therapy and immunotherapy will benefit patients with pancreatic cancers.P < .001) as well as in patients between 50 and 70 . Periodontal disease, tooth loss and root canal infections showed a positive association with an increased risk of developing pancreatic cancer. Previously, Heikkilä et al. (2018), in their register-based cohort study of 68 273 adults, reported a higher pancreatic cancer mortality among individuals with periodontitis . An even stronger association was noted after adjustments . Fan et al. (2018) in their population-based nested case-control study have provided supportive evidence towards the role of oral periodontopathogenic microbiota in the aetiology of pancreatic cancer.Yu et al. (2022) also confirm the association between poor oral health and pancreatic cancer risk. They found a statistically significant association between periodontitis and an increased risk of pancreatic cancer in patients under the age of 50 , Fusobacterium nucleatum and Tannerella forsythia have a more potent role in the causation of cancers of the digestive tract than previously thought. They are able to evade the immune mechanisms and colonise the gastrointestinal tract, disrupting the epithelial integrity. Of these, Td is a notoriously invasive motile anaerobe with an equally harmful surface-bound chymotrypsin-like proteinase  virulence factor. The possible mechanisms by which it can promote carcinogenesis include (a) hydrolysis of bioactive peptides; (b) inhibition of apoptosis; (c) promotion of cellular invasion; (d) degradation of multiple host proteins; and (e) modulate immunity and inflammation.Evidence shows that periodontopathogens like  Females with periodontal disease were reported to be 3 times more likely to develop oesophageal cancer. Hence, periodontal disease could be a significant risk factor for cancer, especially in mature women.Additionally, research has found that certain potent periodontopathobionts are implicated in the tumorigenesis and progression of cancers affecting not only the oral cavity but also oesophageal, breast and gall bladder carcinomas. According to Nwizu et al. (2020), periodontal disease raises the general cancer risk by 14%.Patients with pre-existing gum disease are currently likely to have an increased risk for pancreatic cancer. As periodontal disease is a modifiable risk factor, it is of uppermost importance that regular and valid screening measures are employed for its early detection.9To further add to the knowledge, dentilisin acts against various immunomodulatory proteins critical for the regulation of tumour microenvironment and inflammation, thereby further contributing to tumour progression. It also has the ability to modulate immunomodulatory proteins, including matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs), both of which have been implicated in regulation of tumour microenvironment and metastasis of gastrointestinal cancers.MMPs are derived from host PMNs, endothelial, epithelial and smooth muscle cells in latent pro-MMP form. Of all MMPs, particularly activity of MMP-8 (collagenase) is elevated in patients suffering from gum disease as it causes destruction and digestion of type I collagen which is present in periodontal connective tissue.13 Elevated aMMP-8 levels in mouthrinse and patients’ periodontal disease can be conveniently detected by aMMP-8 POCT  kits readily available in the market.P-8 POCT . These aThe authors recommend utilising non-invasive biomarkers like aMMP-8 for early detection of periodontal disease since it is a risk factor for carcinomas.


Bone is a vital tissue as it carries out various metabolic functions: support of the body, protection of the internal organs, mineral deposit and hematopoietic functions. It has recently been observed that bone tissue also has endocrine functions, producing substances with hormonal activity such as osteocalcin and Fibroblast Growth Factor-23 (FGF-23) ,2.As is well known, bone tissue is continuously renewed through the bone remodelling process. The first phase involves bone resorption, mediated by osteoclasts, and the subsequent phase bone formation, mediated by osteoblasts. The continuous succession of these two phases keeps the skeleton young and allows the repair of microcracks .Bone metabolism, which is finely regulated by different biologic signals ,3, can bThe identification of mechanisms underlying the pathophysiology of metabolic bone diseases continues to be an area of significant research efforts, attracting scientists from different fields.For this Special Issue, we received different scientific contributions, spanning in vitro studies to clinical research articles and reviews of the scientific literature.The paper by Jansen et al., entitled “Increased bone resorption during lactation in pycnodysostosis” , exploreNa et al., in the paper entitled “Aesculetin inhibits osteoclastic bone resorption through blocking ruffled border formation and lysosomal trafficking” , showed Osteoporosis is generally considered more prominent among women than men. However, in the review by Rinonapoli et al., entitled “Osteoporosis in men: a review of an underestimated bone condition” , the autIn the review entitled “Mechanisms of bone fragility: from osteogenesis imperfecta to secondary osteoporosis” , El-GazzRecent research suggests that the microbiota is involved in the regulation of bone metabolism, and that its alteration may induce osteoporosis. This is the topic of the review by De Martinis et al. entitled “The osteoporosis/microbiota linkage: the role of miRNA” . In part


Natural killer (NK) cells are innate immune cells that demonstrate cytolytic activity against tumor cells, virus-infected cells and other physiologically stressed cells, such as senescent cells. The scientific community continues to discover novel biological aspects of NK cells in different organs, tumor microenvironments, and age groups. Recently, cancer immunotherapy has achieved exceptional success. Novel agents and combination therapies that modulate the immune system, including NK cells, provide promising routs for future treatment. International Journal of Molecular Sciences, entitled “Natural Killer Cells and Immunotherapy”, includes eight contributions: five original articles and three reviews that each provide new information on NK cell biology and immunotherapy. This Special Issue of the In their review “The Biological Role and Therapeutic Potential of NK Cells in Hematological and Solid Tumors”, Velichinskii et al.  reviewedIn the review entitled “Postoperative Natural Killer Cell Dysfunction: The Prime Suspect in the Case of Metastasis Following Curative Cancer Surgery”, Market et al.  introducSome immune cells exhibit unexplained features and functions in our immune system. Natural killer T (NKT) cells possess the common features of T and NK cells in terms of their immune responses. In particular, invariant NKT (iNKT) cells, expressing an invariant TCR α-chain, constitute a major population of NKT cells . In the Five original articles introduce the novel findings of NK biology and immunotherapy under different contexts.Cytokines are critical determinants of the phenotypic characteristics and functional activity of NK cells in immune response and development. In the article entitled “Pro- and Anti-Inflammatory Cytokines in the Context of NK Cell-Trophoblast Interactions”, Mikhailova et al.  introducIn the article “NK Cell-Dependent Antibody-Mediated Immunotherapy Is Improved In Vitro and In Vivo When Combined with Agonists for Toll-like Receptor 2 in Head and Neck Cancer Models”, Gruijs et al.  investigIn a short communication article entitled “A Chimeric IL-15/IL-15Rα Molecule Expressed on NFκB-Activated Dendritic Cells Supports Their Capability to Activate Natural Killer Cells”, Bosch et al.  reportedIn the article “Ex Vivo Expanded and Activated Natural Killer Cells Prolong the Overall Survival of Mice with Glioblastoma-like Cell-Derived Tumors”, Shida et al.  reportedIn the final article presented in this publication, “NLRP3 Deficiency in Hepatocellular Carcinoma Enhances Surveillance of NK-92 through a Modulation of MICA/B”, Lee et al. [As described in this Special Issue, NK cells are representative immune effector cells for the development of immunotherapies for hematological and solid cancer treatments. The possibility of using NK cells is growing rapidly, and will open up new opportunities and prospects for the development of immunotherapy in both hematological and solid cancer treatments; however, many questions remain to be elucidated in clinical trials and basic research. The results and discussions of these articles shed considerable light on NK biology and immunotherapy.


Hepatocellular carcinoma (HCC) and biliary tract cancers (BTC) represent the two major forms of primary liver cancers. Despite the growing efforts to translate the increasing knowledge on molecular alterations of these cancers into treatment options for patients, the actual clinical outcomes remain unsatisfying .BTCs are fatal gastrointestinal cancers with very poor 5-year survival rates . The incHCC is the most common form of liver cancer . This deGuo et al.; Shen et al.; Yang et al.) and seven original research papers .Due to the ineffectiveness and development of resistance to current therapies, the need to identify and provide alternative therapeutic approaches is of paramount importance to alleviate the suffering of BTC and HCC patients. Therefore, the current Research Topic provided a structural platform to identify mechanisms of resistance, novel relevant therapeutic targets and prognostic/predictive markers as well as to demonstrate promising innovative treatment options. The call for papers attracted an astonishing number of 10 highly interesting publications and over 65 authors contributed to this Research Topic in the form of three structured reviews , non-invasive preoperative prediction of angiogenesis related markers in BTC , optimizing strategies for immunotherapy and the current status of adjuvant therapies in HCC are discussed , characteristics of extrahepatic cholangiocarcinoma (eCCA) are analyzed , and novel therapeutic strategies for BTC are demonstrated. .This Research Topic covers a variety of subject areas: promising new survival markers after surgery for BTC patients .High levels of vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) correlate with tumor progression and poor prognosis in eCCA patients . HoweverJansson et al.'s study, three preoperative immunologic plasma markers were identified - CSF1, TIE2, and TRAIL - that predict survival after surgery in a cohort of 102 BTC patients utilizing high-throughput multiplex immunoassay. CSF1 and TIE2 were found to be negative prognostic factors in BTC, while TRAIL was demonstrated to be a positive prognostic factor .BTC patients typically experience cancer recurrence within 5 years of surgery, but prognostic factors such as lymph node metastasis and tumor grading can only be observed after tumor resection . In JansShen et al.).Immunotherapy is a therapeutic option for HCC patients; however, the immune microenvironment of many tumors suppresses the effectiveness of this treatment .To address this Research Topic, Liang et al. also identified an efficacy enhancement of immunotherapy in HCC by combining anti-PD-1 antibodies with Abrine, an inhibitor of indoleamine 2,3-dioxygenase 1 (IDO1). In their study, Liang et al. showed that IDO1 is upregulated in HCC cells and can lead to tumor immune escape. They found that using Abrine along with anti-PD-1 antibodies can inhibit immune escape and increase CD8+ T cell infiltration, leading to a stronger immune response and anti-tumor effect .Taken together, this Research Topic provides insight into the latest efforts to overcome resistance mechanisms of current therapies, discover novel prognostic and predictive markers, and identify alternative anti-BTC/HCC strategies.We sincerely thank all the authors for their valuable contributions to this Research Topic.


Molecular biomarkers are emerging as the key indices for the management of patients with significant diseases. Motivated by the systematic effort to define the human genome, the creation of rapid analytic technologies for evaluating nucleic acids and proteins has provided the technological “boom” for the development of molecular biomarkers. Collaboration and cooperation between stakeholders involved in biomarker development, application, and regulation may be the most expeditious route toward the translation of laboratory discovery into patient management. In summary, intensive research has originated multiple factors or biomarkers that are likely to be helpful in diagnosis, characterization, and therapy selection of different patients. A thorough understanding of the relevance of each biomarker will be the key to efficiently diagnose a disease and direct the patients towards the drugs more likely to be of benefit based on their particular profile. The papers selected for this special issue represent an excellent panel for addressing the molecular biomarkers as the future tools of medicine. This special issue contains thirteen papers.Circulating microRNAs and kallikreins before and after radical prostatectomy: are they really prostate cancer markers?,” M. G. Egidi et al. presented 38 patients with prostate cancer. They suggested that two miRNAs (miR-21 and miR-141) could be involved in post surgical inflammatory processes. Postoperative serum kallikreins showed a significant decrease, highlighting the potential usefulness of kallikreins apart from PSA as potential prostate cancer markers.In the paper entitled “A novel differential predict model based on matrixx-assisted laser ionization time of flight mass spectrometry and serum ferritin for acute graft-versus-host disease,”. C.-Y. Zhang et al. investigated the possibility of pre warning the risk of an acute graft-versus-host disease (aGVHD) before and after allogeneic hematopoietic stem cell transplantation  by serum profiling combining with serum ferritinin. Their joint prewarning model could predict the risk of aGVHD, especially severe aGVHD before transplant which provide a reliable method to continuously monitor the condition of patients.In the paper entitled “The use of multidimensional data to identify the molecular biomarker for pancreatic ductal adenocarcinoma,” L. Zhuang et al. presented that they have adopted an integrative approach to simultaneously identify biomarker and generate testable hypothesis from multidimensional omics data. They have found that PER2 expression was highly associated with the survival data, thus representing a novel biomarker for earlier detection of pancreatic ductal adenocarcinoma (PDAC).In the paper entitled “Clinical evaluation and cost-effectiveness analysis of serum tumor markers in lung cancer,” R. Wang et al. showed that combinations of four tumor markers  improved the sensitivity for lung cancer and different combination panels had their own usefulness. NSE, CEA, and CYFRA21-1 were the optimal combination panel with highest Youden's index (0.64), higher sensitivity (75.76%), and specificity (88.57%), which can aid in clinical diagnosis of lung cancer.In the paper entitled “Immune parameters in the prognosis and therapy monitoring of cutaneous melanoma patients: experience, role, and limitations,” M. Neagu et al. reported the follow-up for 36 months of the immune parameters of patients diagnosed in stages I–IV, namely, pre- and postsurgery immune circulating peripheral cells and circulating intercommunicating cytokines.In the paper entitled “Comparative gene expression profiling in human cumulus cells according to ovarian gonadotropin treatments,” S. Assou et al. provided an exclusive study characterizing gene expression profiles in cumulus cells (CCs) of periovulatory follicles from patients undergoing HP-hMG and rFSH gonadotropin treatments during in vitro fertilization cycles. This project has characterized the expression of these genes as biomarkers of in vitro embryo quality.In the paper entitled “Aptamers: novel molecules as diagnostic markers in bacterial and viral infections?,” F. M. Zimbres et al. urged an urgent need to discover novel diagnostic as well as therapeutic tools against infectious agents. They viewed that the systematic evolution of ligands by exponential enrichment (SELEX) represents a powerful technology to target selective pathogenic factors as well as entire bacteria or viruses. SELEX uses a large combinatorial oligonucleic acid library (DNA or RNA) which is processed by a high-flux in vitro screen of iterative cycles.In the paper entitled “Distribution of ABO blood group and major cardiovascular risk factors with coronary heart disease,” S. Biswas et al. viewed that the AB blood group decreases the risk of CHD in healthy controls; it might be due to the higher concentration of high density lipoprotein cholesterol (HDL-c), while the O blood group increases the risk of CHD due to lower HDL-c levels in Bengali population of eastern part of India.In the paper entitled “Immunomodulatory effect of continuous veno-venous hemofiltration during sepsis: preliminary data,” G. Servillo et al. reported that severe sepsis and septic shock are the primary causes of multiple organ dysfunction syndrome (MODS), which is the most frequent cause of death in intensive care unit patients. Many pro- and anti-inflammatory mediators, such as interleukin-6 (IL-6), play a key role in septic syndrome. Continuous renal replacement therapy (CRRT) removes in a nonselective way pro- and anti-inflammatory mediators. The authors investigate the effects of continuous venovenous hemofiltration (CVVH) as immunomodulatory treatment of sepsis in a prospective clinical study.In the paper entitled “Acetylcholinesterase as biomarker in environmental and occupational medicine: new insights and future perspectives,” M. G. Lionetto et al. viewed that Acetylcholinesterase (AChE) is a key enzyme in the nervous system (NS), since it terminates nerve impulses by catalyzing the hydrolysis of acetylcholine. As a specific molecular target of organophosphate and carbamate pesticides, AChE activity and inhibition are a human biological marker of pesticide poisoning. Thus, it is used to study the effects of the exposure to organophosphate and carbamate pesticides on NS in occupational and environmental medicine. This paper reviews and discusses the recent findings about AChE, including its sensitivity to other pollutants and expression of different splice variants. These insights open new perspectives for the use of this biomarker in environmental and occupational human health monitoring.In the paper entitled “Polyisoprenylated methylated protein methyl esterase is both sensitive to curcumin and overexpressed in colorectal cancer: implications for chemoprevention and treatment,” F. Amissah et al. discussed polyisoprenylated methylated protein methyl esterase (PMPMEase) which cocatalyzes the polyisoprenylation pathway required to process various monomeric G proteins. Mutation of these G proteins is considered to be responsible for 50% of colorectal cancers. This interesting finding suggests that elevated PMPMEase activity and its overexpression can be one of the candidate markers for early diagnosis of colorectal cancer. Susceptibility of this enzyme to curcumin also suggests that PMPMEase can be a potential candidate for targeted anticancer therapy.In the paper entitled “Emerging therapeutic biomarkers in endometrial cancer,” P. Dong et al. reviewed the current status of molecular therapies tested in clinical trials and mainly discussed the potential therapeutic candidates that are possibly used to develop more effective and specific therapies against endometrial cancer progression and metastasis.In the paper, entitled “Mouse prostate epithelial luminal cells lineages originate in the basal layer where the primitive stem/early progenitor cells reside: implications for identifying prostate cancer stem cells” J. Zhou et al. have developed an in vivo cell fate tracing mouse model and an in vivo slow-cycling cell label mouse model to provide further insight into this question. Through genetic manipulation in the animals, their findings indicate that the basal cell lineage can produce more differentiated luminal cells; the putative mouse prostate stem cells (which are slow-cycling and responsible for tissue maintenance) likely reside in the basal layer.In the paper entitled “ Though the selected topics and papers are not an exhaustive representation of the entire area of molecular biomarkers and tools of medicine, yet they represent the rich and many-faceted knowledge that we have the privilege of sharing with the readers.


The growing concern about cannabis use, the most commonly used illicit drug worldwide, has led to a significant increase in the number of human studies using neuroimaging techniques to determine the effect of cannabis on brain structure and function. We conducted a systematic review to assess the evidence of the impact of chronic cannabis use on brain structure and function in adults and adolescents.Papers published until August 2012 were included from EMBASE, Medline, PubMed and LILACS databases following a comprehensive search strategy and pre-determined set of criteria for article selection. Only neuroimaging studies involving chronic cannabis users with a matched control group were considered.One hundred and forty-two studies were identified, of which 43 met the established criteria. Eight studies were in adolescent population. Neuroimaging studies provide evidence of morphological brain alterations in both population groups, particularly in the medial temporal and frontal cortices, as well as the cerebellum. These effects may be related to the amount of cannabis exposure. Functional neuroimaging studies suggest different patterns of resting global and brain activity during the performance of several cognitive tasks both in adolescents and adults, which may indicate compensatory effects in response to chronic cannabis exposure.However, the results pointed out methodological limitations of the work conducted to date and considerable heterogeneity in the findings.Chronic cannabis use may alter brain structure and function in adult and adolescent population. Further studies should consider the use of convergent methodology, prospective large samples involving adolescent to adulthood subjects, and data-sharing initiatives. Cannabis is the illicit drug most widely available and used worldwide Animal studies have consistently demonstrated that delta-9-tetrahydrocannabinol (THC), the main psychoactive component of cannabis It is remarkable to note that although the onset of cannabis use is typically during adolescence, a few imaging studies have been conducted with adolescent users The growing concern about cannabis use has led to a significant increase in the number of human studies using neuroimaging techniques to determine the effect of the substance on brain structure and function, as well as to several recent reviews examining this topic Data for this systematic review was collected with an advanced document protocol in accordance with the PRISMA guidelines Electronic searches were performed using EMBASE (1980-August 2012), Medline (1966-August 2012), PubMed (1966-August 2012) and LILACS (1982-August 2012) databases. The following key words were used: cannabis; marijuana; marihuana; delta-9-tetrahydrocannabinol; THC; cannabidiol, CBD; neuroimaging; brain imaging; computerized tomography, CT; magnetic resonance, MRI; single photon emission tomography, SPECT; functional magnetic resonance, fMRI; positron emission tomography, PET; diffusion tensor MRI, DTI-MRI; spectroscopy, MRS. All the studies published up to August 2012 were included without language restriction.A general review of all neuroimaging studies investigating brain structure or function was initially performed. We obtained a total of 142 published papers . StudiesWe defined chronic cannabis users as persons who used cannabis several times a week and who had done so for at least two years. Recreational  cannabis users were defined as persons who had used cannabis sporadically (less than four times a month), and naïve users or healthy controls were persons who had used cannabis less than 15 times in their lifetime, according to standardized strict criteria Any publication that reported data using two different neuroimaging techniques from the same subjects  or a study examining the same subjects with two different cognitive tasks  was considered as two studies in this review.Data was independently extracted by two reviewers. In case of disagreement, opinion from a third senior researcher was sought to assess whether study criteria were fulfilled. From the articles included we recorded names of authors, year of publication, socio-demographic  and cannabis use characteristics , imaging type and design, exclusion criteria , confirmation of abstinence from other drugs (whether checked by urine test), rest/active condition , type of cognitive task performed during functional imaging and psychopathological variables assessed . With regard to alcohol use, we assessed if subjects met criteria for alcohol abuse or for excessive alcohol consumption  based on the reported data. For structural and functional imaging data, the primary measures of interest were global and regional volume, and global and regional activity . The secondary outcome was its correlation with clinical variables. We collected the statistically significant results of each outcome variable, and recorded whether a multiple comparison correction was done to prevent bias towards false positives.a priori selection criteria Of the 142 studies identified, thirty-six did not meet the We identified 11 structural MRI studies that examined adult chronic cannabis users and met our selection criteria . StructuOf the seven studies comparing global brain volume measures between chronic cannabis users and healthy controls, there was only one study reporting significant differences et al. (2005) et al. (2011) et al. (2005) Among the six studies employing a whole-brain analysis approach With regard to the three studies that focused on specific regions of interest, all studies reported bilateral volumetric reductions in the hippocampus et al. (2011) et al. (2005) With respect to other brain regions, Cousijn et al. (2008) et al. (2011) et al. (2011) Four studies have used DTI to examine the integrity of white matter tracts in chronic cannabis users et al.  et al. (2011) Three volumetric studies in adolescent chronic cannabis users were included, two of which consist of the same sample et al. (2010) et al. (2011) et al. (2009) In terms of correlations, Medina 15O-PET 133Xe-SPECT 18F-FDG-PET 11C]- raclopride-PET 18F]FMPEP-d2 11C]-raclopride-PET studies et al. (2000) et al. (2001) et al. (2008) 11C]-raclopride-PET binding potential and glucose metabolism 11C]- raclopride-PET studies et al. (2012) et al. (2012) et al. (2011) We included eight case-control studies comparing resting rCBF in adult chronic cannabis users and non cannabis using healthy controls . The imaWe identified 16 studies in adult chronic cannabis users that compared regional activation during the performance of a cognitive task with healthy controls , four wiet al. (2006) et al. (2010) et al. (2012) Chang 15O-PET study, Block et al. (2002) et al. (2007) et al. (2006) In a H2et al. (2004) et al. (2005) et al. (2005) et al. (2004) 15O-PET during the performance of a modified Stroop test. However, they also reported a reduced dorsolateral prefrontal cortex activation and a greater activation in the hippocampus bilaterally et al. (2009) Eldreth et al. (2005) et al. (2011) 15O-PET, and Wesley et al. (2011) et al. (2005) et al. (2011) et al. (2011) et al. (2011) et al. (2010) et al. (2010) et al. (2010) et al. (2010) et al. (2010) Bolla et al. (2011) King et al. (2009) Gruber We included five case-control fMRI studies in adolescent cannabis users comparing brain activity with healthy controls during a cognitive task performance. As an exception, two of them et al. (2007) et al.  et al. (2007) et al. (2008) et al. (2010) et al. (2011) Padula et al. (2007) In a group of abstinent cannabis users, Tapert 1-rich areas implicated in several cognitive functions, which may be related to the amount of cannabis use; (2) Altered neural activity during resting state and under several different types of cognitive paradigms, that may reflect a different recruitment of brain areas during the tasks, particularly within the prefrontal cortex; and (3) The few studies conducted in adolescents suggest that both structural and functional alterations may appear soon after starting the drug use and may be related to gender.In this systematic review, we identified 43 studies suitable for inclusion regarding the impact of chronic cannabis use on brain structure and functioning, of which eight (19%) were in the adolescent population. Despite the high degree of heterogeneity among the studies reviewed herein, several relatively consistent findings emerged from this review. These findings, discussed in detail below, include: (1) Structural brain abnormalities, mainly in CBIn terms of structural findings, specific regional brain analyses demonstrated evidence of structural abnormalities when adult chronic cannabis users were compared with healthy controls. The most consistently reported brain alteration was reduced hippocampal volume Functional neuroimaging studies that have evaluated the resting state in active and abstinent adult chronic cannabis users suggest that resting global Functional imaging studies comparing activation in both adult and adolescent chronic cannabis users with healthy controls during the performance of different cognitive tasks indicated that chronic cannabis users would use similar brain areas that engage these cognitive processes but often demonstrating an altered pattern of brain activity A further important issue emerging out of this review is that few studies have investigated the effects of chronic cannabis use on the brain in adolescence subjects. In light of the popularity of cannabis among teenagers Results presented here have pointed out some important methodological differences that limit the generalisation of results and comparison between studies and have doubtless contributed to the slightly disparate array of findings. Despite the use of a strict definition of chronic cannabis user and robust application of inclusion and exclusion criteria in an attempt to avoid excessive heterogeneity between samples, studies often diverged on certain socio-demographic characteristics and cannabis use parameters, such as gender-bias, age of onset, lifetime use and abstinence period before the acquisition of imaging data. Moreover, it is well known that the THC content of smoked cannabis varies markedly between sources and preparations, with potency reported to have increased substantially over the past ten years With regard to other methodological limitations, some studies have reported modest sample sizes, sometimes below the threshold that would be currently regarded as acceptable  Table S1Click here for additional data file.


The seventh annual Midsouth Computational Biology and Bioinformatics (MCBIOS) conference took place February 19 and 20, 2010 at Arkansas State University in Jonesboro, AR, presided over by Daniel Berleant, this year’s President of MCBIOS.  Keynote speakers were Elaine Ostrander of NIH, renowned for her work on dog genomics, Clayton Naeve, CIO of St. Jude Children's Research Hospital, and Robert Cottingham, Leader of the Computational Biology and Bioinformatics Group, Oak Ridge National Laboratory. The record-breaking attendance exceeded 200 participants which necessitated parallel talk sessions for the first time. Student oral presentation award winners were Heidi Pagan (first place) of Mississippi State University (MSU), Juliet Tang (second place) of MSU, and Aleksandra Markovets (third) of Mississippi Valley State University. In addition, a record number of posters were also presented. Due to the number of posters, student poster awards were given in two categories, Biological Focus and Computational Focus. Winners were N. Platt and V. Chaitankar (1st place), G. Cooper and L. Pillai (2nd), and M. Ammari and C. Gearheart (3rd). Reflecting the integration of these foci in the field, winners in one category frequently scored high in the other category as well. MCBIOS is also pleased to have this year acquired legal status as a non-profit organization.A total of twenty eight papers describing research presented at the 2010 conference were accepted for publication in these proceedings out of a total of forty three papers submitted for consideration (~65%). The number of papers submitted for consideration was the largest submitted since the inception of MCBIOS with a 34% increase over last year’s proceedings , and theet al. describe an expansion of the FDA-developed, ArrayTrackTM genomics tool, into a platform that supports microbial data to enable the rapid detection of food borne pathogenic bacteria during outbreak scenarios [et al. describe new SNP (single nucleotide polymorphism) and QTL (quantitative trait locus) libraries for the ArrayTrack system which provide users with extensive data including links to other resources, and motivated in part by the need to augment gene association study results with biological context [et al. report on the new web-based tool, IView, which graphically displays and makes searchable introgression data by marker name, chromosome number, or map position [Bioinformatics was largely born as a field through the need to analyze and understand sequence data. Partly as a result of ultra-high-throughput sequencing technology, the field has seen a strong resurgence recently with new computational methods being developed and applied to downstream applications such as the identification of human mutations -25 as wecenarios .  Xu et  context . Christoposition .Quest and colleagues working in the Cottingham laboratory address the problem of brittle genome annotation systems with an OWL-based approach that provides several significant advantages . Work reAn examination of substitution rates over evolutionary time by Ulrich Melcher compared viral and non-viral sequences to reveal that the rate of nucleotide interchanges in plant viruses is not constant, impacting phylogeny studies.  The evidence suggests that different evolutionary rules may apply to recent divergences than to those linked to distant speciation events .  et al. using a proteomics approach to evaluate the host protein expression differences. Two gene sets were found, each specific to one disease form which showed significant functional differences [et al. address the need for additional genomic information for Northern bobwhite, an avian species critical to ecosystem maintenance. Using next generation sequencing of cDNAs from a multi-tissue library, they generated ESTs which were pipelined into a unigene database and made publicly available at a searchable website [An in depth analysis of the two forms of Bovine Viral Diarrhea, a worldwide cattle disease, was completed by Mais Ammari ferences .  Arun R website .  et al [et al [Arabidopsis. The conserved gene evolution indicated that the study in the model species can be translational to human disease studies. Rowena Kelly et al [Aspergillus flavus resistance in Maize such as gene expression, proteomic, QTL genetic mapping studies and sequence data from the literature to help identify and select candidate A. flavus resistance genes.Cyriac Kandoth et al  mined thl [et al  carried ly et al  have conBiological information may be encoded in a one-dimensional sequence, but its manifestation takes place in three-dimensional space in the form of proteins, cells and tissues among other structures. Computational analysis and prediction of these structures is challenging, to say the least, and an active area of recent bioinformatics interest -48. Compet al. report on a method for modeling interactions between 8R-lipoxygenase and its substrate [et al., which can serve as an important training and practice tool for surgeons [et al.[To better understand and improve of the creation of anti-inflammatory drugs through inhibition of lipoxygenase, an essential enzyme in the inflammatory pathway, Shuju Bai ubstrate . A mixedsurgeons .  The pos [et al..  They ret al [et al [et al  compared density based clustering and Fuzzy C-Means clustering algorithms for border detection in dermoscopy images finding that density based clustering performed best with a low border error, high precision and recall; however, the Fuzzy C-Means clustering algorithm had poor performance especially in border detection.  Mete et al  proposed two new methods, boundary driven density based clustering based algorithms which performed better delineation with noisy images and an active contour model that gave the best detection with optimum parameters. Denise Koessler et al [Sinan Kockara et al  and Mutll [et al  examineder et al  developeThere are many areas within biology that are amenable to computational analysis, each of which is usually approached separately. Somewhat recently, the term “systems biology” has emerged to describe interconnected analyses, particularly those that help reduce complexity in these systems, as defined by the number of interconnections between component parts, into a more functional understanding . The term “systems biology” is often used broadly and not always consistently, but is an emergent area of high interest in bioinformatics -57. et al.[et al. report on the development of a Web-based gene-discovery bioinformatics tool, FAUN (Feature Annotation Using Nonnegative matrix factorization), which identifies implied gene relationships from biomedical literature, enabling hypotheses on functional characterization [A novel method for identifying subnetwork markers in a human protein-protein interaction network is described by Junjie Su et al..  They rrization .  et al., explores the important problem of inferring gene regulatory networks from time sliced microarray data. They claim that mutual information (MI) based approaches have inherent limits to their capabilities in this context which they were able to improve upon with algorithms based on modifications to the mutual information metric [ A paper by Vijender Chaitankar, n metric .Kumar and Nanduri present a downloadable database, HPIDB, which serves as a resource of host-pathogen protein-protein interactions integrated non-redundantly from public databases. The authors report on the flexibility of the database for querying and an output format which allows portage to downstream applications .Microarrays have long been a subject of bioinformatics analysis not only for better understanding transcription, a process affected by both genetic and epigenetic factors ,63, but A new non-stationary Dynamic Bayesian Network with a flexible lag choosing mechanism that detects potential genetic regulators and has improved structure prediction is reported by Yi Jia and Jun Huan .et al. address two interesting QC questions of whether expired Affymetrix GeneChip microarrays, an expensive resource, are still useful up to four years after expiration and if RNA stored at -80°C for the same period was suitable as template source.  In spite of a decrease in sensitivity, the authors found that these arrays generated data consistent with that from unexpired arrays and report favorably on the stability of the RNA [Zhining Wen  the RNA .There are a few other papers published in the proceedings but, for space, are not summarized here: -69.http://www.ISCB.org). For information regarding MCBIOS and our future meetings, see http://www.MCBIOS.org.MCBIOS 2011 will be held in College Station, Texas.  The 2010-2011 President of MCBIOS is Ulisses Braga-Neto of the University of Texas A&M, and Susan Bridges of Mississippi State University is now the President-elect. MCBIOS is a regional affiliate of the International Society for Computational Biology (The authors have no competing interests to declare.All authors served as editors for these proceedings, with JDW serving as Senior Editor. All authors helped write this editorial.


Transcranial direct current stimulation (tDCS) is a noninvasive neuromodulatory technique with putative cognitive enhancing and therapeutic applications. Since the year 2000, almost 1000 papers have been published on tDCS, reflecting the possible significance of a cheap, safe, and easily applied neuromodulatory technology. Whether or not this potential is tapped depends on understanding how tDCS affects brain functioning, a question explored in a recent publication by Stagg et al. ; presenttDCS is thought to decrease neuronal resting membrane potential beneath the anodal electrode by pumping in positive ions; vice versa for cathodal stimulation  underwent two tDCS sessions  in a counterbalanced order, separated by a week. tDCS electrodes were applied to left DLPFC and the contralateral supraorbit. In each session, participants underwent a 40-min resting-state ASL functional MRI scan in which there was a 10-min baseline, 20-min of concurrent tDCS-ASL and a 10-min post-stimulation period. Separately, but using the same protocol, a second group of participants (N = 12) were scanned once under sham stimulation only. Stagg et al. compared resting-state CBF between these three periods and examined changes in functional connectivity between the regions beneath each electrode and the rest of the brain. The same analyses were conducted using the sham stimulation group to confirm that the changes were not driven by nonspecific temporal drifts.Stagg et al. studied the effect of tDCS on cerebral perfusion using ASL. Subjects  to group 2 (stimulation—baseline) should be conducted; finding a statistically significant effect in one sample, but not another is not a valid approach to establish the between-group effect (Nieuwenhuis et al., The electrical current used in tDCS studies is insufficient to generate neuronal action potentials. Instead, spontaneous firing changes resulting from alterations in neuronal resting membrane potential are thought to underlie the neuromodulatory effects of tDCS (Bikson et al., While the clinical potential for tDCS is high, it is obscured by our lack of insight into its neural effects. Studies such as Stagg and colleagues' build on previous findings and pave the way to an improved understanding. Only with this comprehension can the full clinical possibility of tDCS be utilized and directed toward improving treatment.Camilla L. Nord, Níall Lally, and Caroline J. Charpentier wrote the manuscript together.


Frontiers in Fungi and their Interactions details the exciting progress in developing vaccines and immunotherapy for fungi.Invasive fungal diseases have increased many fold over the past 50 years. Current treatment regimens typically require prolonged administration of antifungal medications that can have significant toxicity. Moreover, our present potent antifungal armamentarium fails to eradicate fungal pathogens from certain compromised hosts. Additionally, invasive fungal diseases continue to have unacceptably high mortality rates. A growing body of work has focused on the utility of vaccines and/or immunotherapy as a powerful tool in combating mycoses, either for the active treatment, as an adjuvant, or in the prevention of specific fungal pathogens. This Research Topic “Vaccines and Immunotherapy against fungi: a new frontier” in Candida albicans (Vecchiarelli et al., Aspergillus fumigatus (Diaz-Arevalo et al., Cryptococcus neoformans (Hole and Wormley, Paracoccidioides brasiliensis (Travassos and Taborda, P. brasiliensis is also presented (Mayorga et al., Artocarpus heterophyllus, which modulates immunity against P. brasiliensis (Ruas et al., Histoplasma capsulatum (Nosanchuk et al., The critical requirement for understanding the degrees of engagement of host defense pathways in responding to fungal invasion has led to an increased focus on host-pathogen interactions. In this Research Topic, Carvalho et al.  review oIn sum, these articles broadly paint the current spectrum of investigations on host-pathogen interactions and provide a review of the state-of-the-art in vaccinology and immunotherapy against fungi. The information presented also underscores the rich areas for future study, all promising improved therapeutics against fungal invaders.


APP are linked to early onset of familial Alzheimer’s disease (FAD); APP processing generates β-amyloid (Aβ) peptides, which are the principal components of the amyloid pathology. Therefore, understanding the role of APP in neuronal function and dysfunction will provide crucial insights to AD pathogenesis.Genetic and biochemical studies establish a central role of the amyloid precursor protein (APP) in Alzheimer’s disease (AD): Genetic mutations and gene amplification of We have a long-standing interest in studying the physiological functions of APP in neurons and synapses. Analysis of various loss-of-function mutants of APP and combining with a mixed-culture system allowed us to identify APP as a synaptic adhesion protein. We recently uncovered a potent role of APP in mediating adult neurogenesis in dentate gyrus: Loss of APP results in an aberrant increase in progenitor proliferation but impaired newborn neuron differentation, maturation and integration. Intriguingly, we found that APP is highly expressed in GABAergic interneurons and that specific deletion of APP in these neurons, but not in excitatory neurons, leads to similar neurogenesis defect. Our mechanistic and functional studies indicate that this activity is mediated by a general role of APP in regulating GABA release through its synaptic adhesion property.AD is an age-related disease. Adult neurogenesis declines sharply with aging. Increasing evidence supports a role of hippocampal adult neurogenesis in brain function and its impairment in AD. Therefore, perturbation of APP-mediated adult neurogenesis may contribute to neuronal dysfunction and AD pathogenesis.


It is unique in its strict selection of a single member for activation, a process termed monoallelic expression. The conceptual advances that have emerged from studying var genes show striking common epigenetic features with many other clonally variant gene families or even single-copy genes that show a variegated expression in parasite populations. However, major mechanistic questions, such as the existence of a potential expression site and the identity of transcription factors or genetic elements driving singular gene choice, are still unanswered. In this review we discuss the recent findings in the molecular processes essential for clonal variation, namely silencing, activation, poising and switching. Integrating findings about all clonally variant gene families and other mutually exclusive expression systems will hopefully drive mechanistic understanding of antigenic variation.Phenotypic variation in genetically identical malaria parasites is an emerging topic. Although antigenic variation is only part of a more global parasite strategy to create adaptation through epigenetically controlled transcriptional variability, it is the central mechanism enabling immune evasion and promoting pathogenesis. The  The sites of host–parasite interaction are constantly exposed to recognition by the host immune system. Protozoan pathogens have developed a wide range of sophisticated immune evasion strategies such as antigenic variation . Most var genes are in subtelomeric regions, whereas others are arranged in more chromosome central positions. Var genes consist of an Exon1, coding polymorphic sequences forming the extracellular domain, an Exon2, coding the semi-conserved intracellular domain, connected by a single highly conserved intron.The protozoan pathogen d stream A. Duringvar gene expression studies have wide implications for other plasmodial gene families and single-copy genes controlled by similar epigenetic mechanisms  A and B subtypes, as well as some members of the rifins, another clonally variant gene family. Inactivation of a second member of the Sir2 family, called Sir2B, showed some complementary de-repression effect on other, mostly upsC, var gene members , which is enriched in promoter regions of repressed var genes  to silent but not active P. falciparum than was anticipated   and Histone 3 lysine 9 acetylation (H3K9ac), which renders it permissive for transcription   in var gene is active within a single cell. The HB3 P. falciparum strain contains two nearly identical copies of the var2csa gene. Despite the fact that one copy lies on chromosome 12 and the other on chromosome 1, RNA FISH analysis showed that simultaneous expression of both copies occurs at the same site in the nucleus . Interestingly, an episomal promoter of the rifin gene families also colocalizes with the active var gene expression site when activated through drug selection . An alternative view is that an activation complex is recruited to the var gene in situ. This initiates the transcriptional activation cascade including its relocation away from the repressive zone. This scenario predicts that any perinuclear site outside the repressive centres may form an expression site. Depending on which model turns out to be accurate, var gene activation would occur before segregation from the repressive cluster or segregation would be a precondition for activation. In either case, the recent discovery of nuclear actin associated to var gene intron regions opens a new exciting avenue. Nuclear actin could provide a mechanical framework for spatial organization of active and silent genes .A different hypothesis for a mutual exclusive expression mechanism postulates the existence of a unique Var genes are only transcribed during the early stages of the blood stage cycle and intrinsic switching rates determined from in vitro cultured parasites can reach up to 2% per cycle (Roberts et al., var genes will be re-activated after the next round of invasion (var promoter in the late blood cell stages, while H3K9me3 is prevented from spreading into the promoter region (Lopez-Rubio et al., var gene locus in post-ring stages, whereas repressing marks are excluded from this site, presumably contributing to the poised state (Volz et al., P. falciparum awaits further investigation.invasion A. ChIP aSwitching between the clonally variant genes is central to antigenic variation and must be adapted to the host so the variant gene repertoire is not depleted too fast whilst effective immune evasion is still possible. It is important to keep in mind that the parasite needs to control its own proliferation so the host is not severely harmed by high parasitemia and effective transmission is guaranteed.var members (Horrocks et al., var genes during the non-transcribed phase can be experimentally erased by promoter titration (Dzikowski and Deitsch, et al., var genes is independent of previous stages. Another recent study on switch pathways used experimental evidence combined with mathematical modelling to conclude that var switching is non-random and necessitates a balanced process of parasite-intrinsic switching and immune-mediated selection (Recker et al., var gene switching patterns using silencing with drug-selectable markers in multiple isolates suggests that a var gene switching pattern is conserved throughout different genetic backgrounds (Enderes et al., et al., To this day, no genes modifying switch rates have been identified and no functional data is available on the molecular mechanism of switching. Nevertheless, descriptive studies demonstrated that transition rates can vary between var gene silencing may be modulated by external factors. For example, Sir2 depends on NAD levels for its activity. Low nutrition states of patients may influence Sir2 activity and hence alter the switch rate of distinct var subtypes. Experimental data from in vitro cultured parasites using peroxide or starvation stress can lead to a slight up-regulation of central var genes (Rosenberg et al., var gene expression pattern in patients (Merrick et al., Another critical question is whether the observed intrinsic switch rate is influenced by external factors, i.e. does the parasite react to its environment, or is the switching pattern hard-wired into the parasite's genome? Although no longitudinal data from human patients are available to support this concept directly, the epigenetic machinery controlling var expression studies from malaria patients, however, support the idea that host factors contribute to var gene transcriptional control. Unlike in vitro cultured parasites, ups A-type var genes were frequently expressed in P. falciparum-infected patients (Bachmann et al., et al., var genes mostly of the ups B and C-type. It will be crucial to investigate whether host factors modify the intrinsic switch rate of ups A-type var genes.The relative paucity of patient data related to this topic makes it difficult to investigate switching rates in the human host. Two recently published var genes cooperate with chromatin modifying enzymes in a complex interplay. On top of this, spatial regulation creates specific nuclear sub-compartments most likely critical for default silencing and monoallelic expression. Other factors such as non-coding RNA produced in subtelomeric regions adjacent or within var genes may contribute as well to antigenic variation (Epp et al., et al., Mutually exclusive expression relies on distinct layers of genetic and epigenetic control factors. Genetic elements, such as the promoter and the intron of var gene expression site remain to be explored. The biggest puzzle, however, in understanding mutual exclusive expression, is the absence of a putative activation factor. The concept of a limiting factor driving monoallelic var transcription has been postulated on many occasions but remains to be demonstrated. The notion that a unique enhancer element is the activating factor or expression site body would seem plausible, since there is exactly one copy in the nucleus of this haploid parasite. A transcription factor, whose expression is restricted to extremely low levels, may be considered but it seems difficult to really make sure that exactly one transcription factor is present. Considering that under special conditions more than one promoter can be activated we favour a model where potential positive feedback loops within the activating complex could ensure aggregation around the expressed var gene. In the absence of candidate genes for a monoallelic activating factor, identification of mutant parasites that have lost var gene expression remains the biggest challenge in the field. Traditional biochemical and mass spectrometric analysis of factors interacting with key genetic elements (Zhang et al., et al., et al., et al., Properties and composition of the hypothetical It remains enigmatic how one gene is active while others are silenced, when and how a specific gene is poised rather switched, but many exciting research avenues are left to explore in the field of mutually exclusive expression.


Dear Editor,In your June 2011 issue, we found an interesting study by Farzanegan et al. entitled, “Effects of lumbar discectomy on disability and depression in patients with chronic low-back pain.” , 4. DeprThe association between depression and pain after low-back surgery is not consistent across studies. Depression and disability have been found to be highly influential in patients undergoing lumbar spine surgery. The psychological profile of depression has predicted poorer outcomes in patients with chronic sciatica pain and disability . The stu


In contrast to the current therapy, which antagonises the formation of proinflammatory mediators, the “proresolving” therapy promotes natural antiinflammatory processes. It is likely that several drugs and phytochemicals would act in this way, but this point has not been investigated and thus might be totally overlooked. In this paper, effects of curcumin (diferuloylmethane) were analysed, considering the ability of this natural compound to affect resolution of inflammation through modulation of its important inputs – activity and apoptosis of neutrophils. The presented data indicate that, besides its well-known ability to suppress mechanisms engaged at the onset and progression of inflammation, curcumin could support resolution of inflammation through decreased activity and enhanced apoptosis of neutrophils. This substance decreased the formation of oxidants in neutrophils, both under in vitro conditions and after oral administration to arthritic rats. Moreover, curcumin accelerated spontaneous apoptosis of neutrophils, as indicated by increased externalisation of phosphatidylserine, by intercalation of propidium iodide and by enhanced activity of the executioner caspase-3.Novel strategies of antiinflammatory therapy are based upon pharmacological agents capable to enhance the resolution –  During the inflammatory process, different cytokines, growth, transforming and chemotactic factors are synthesised and several types of cells become activated. Neutrophils (polymorphonuclear leukocytes) are considered to be central cells of acute inflammation. These cells most rapidly reach the site of injury or infection and liberate antimicrobial proteins, proteases and produce reactive oxygen species. Prolonged or excessive liberation of these very effective and toxic substances could intensify the inflammatory process and enhance tissue damage. Therefore activity of neutrophils declines consecutively by the effect of antiinflammatory mediators  and programmed death of these cells (apoptosis) is initiated. These processes are essential for the termination (resolution) of the beneficial inflammation , luminol, isoluminol and PMA (phorbol-myristate-acetate) were obtained from Sigma  and methotrexate was from Pliva-Lachema . Human Annexin V/FITC Kit was purchased from Bender MedSystems , Caspase-Glo 3/7 Assay from Promega  and human purified caspase-3 was from Enzo Life Sciences .in vitro conditions, isolated human neutrophils were stimulated with 0.05 µM PMA and chemiluminescence was enhanced either with isoluminol (extracellular) or with luminol in the presence of extracellular scavengers – superoxide dismutase and catalase  and propidium iodide. Only Annexin positive cells were considered pre-apoptotic cells and double positive cells (Annexin+/propidium+) were considered late-apoptotic or dead cells.et al., Effect of curcumin on caspase-3 activity was evaluated in cell-free system by the luminescence method, using Caspase-Glo 3/7 Assay kit and human purified caspase-3  elevated the percentage of preapoptotic and apoptotic neutrophils A and incet al., 2, myeloperoxidase, collagenase, as well as to its ability to modulate activities of T lymphocytes and macrophages  were reduced more effectively than oxidants formed within neutrophils (involved in elimination of phagocytosed pathogens and fulfilling a regulatory role). Compared to previous studies (Limasset in vivo conditions – in arthritic rats. Neutrophils modified by inflammation produced several times more oxidants than did healthy controls and this hyperreactivity was suppressed by orally administered curcumin. Since the excessive formation of radicals was ascribed to the enhanced activity of NADPH oxidase in the presence of cytokines (Fairhurst et al., et al., et al., et al., et al., The decreasing effect of curcumin on neutrophil activity was further observed under et al., Curcumin-related inhibition of neutrophil oxidative burst was comparable with the activity of methotrexate – a drug widely used in the therapy of arthritic patients, which triggers synthesis of the endogenous anti-inflammatory mediator adenosine (Cronstein, The presented data indicate that, besides its ability to suppress mechanisms engaged at the onset and progression of inflammation, curcumin could support resolution of inflammation through decreased activity and enhanced apoptosis of neutrophils.


Induction of premature (stress induced) cellular senescence of normal cells provides the mechanism eliminating proliferative capability of potentially carcinogenic cells and thereby is considered to be a barrier preventing neoplastic tranformation . On the Additional evidence on the role of perturbed dNTP pools in induction of DNA damage and replication stress was provided by Bester et al.,. Their sG12V, a classical model of oncogene-induced senescence. Among the enzymes down-regulated by activated HRAS were TS, ribonucleotide reductase subunits M1 and M2 (RRM1 and RRM2). Ectopic coexpression of TS, RRM1 and RRM2 or addition of deoxyribonucleosides to a large extent prevented DNA damage, decreased expression of senescence-associated phenotypes as well as attenuated the extent of proliferation arrest. Interestingly, overexpression of TS, RRM1 and RRM2 in quiescent NHFs had no effect on their proliferation arrest suggesting that unlike the OIS, quiescence is not induced by perturbation of dNTP pools. Moreover, in a separate study, Mannava et al.,[In the subsequent studies Mannava et al., provideda et al.,demonstraMost of the above findings were reproduced in a study of Aird et al., on a rolThe seminal findings on different cellular and oncogene models that perturbation of dNTP pools leads to DNA damage, replication stress, irreversible senescence -8,may ha


Cardiac hypertrophy is a distinguished feature of several physiological and pathological remodeling (Frey et al., Distinct underlying signaling pathways for physiological and pathological hypertrophy have been identified (Maillet et al., Unlike pathological hypertrophy, only a little studies described how miRNAs response to physiological hypertrophy (Soci et al., Traditionally, the adult mammalian heart is recognized as a post-mitotic organ with no regenerative capacity for cardiomyogenesis (Rosenzweig, With the strategies for checking myocyte hypertrophy and new cardiomyocyte formation, the miRNA basis of physiological hypertrophy will be revealed, which will help develop a miRNA-based therapy for heart failure.


MAOA, COMT, and 5HTTLPR each have predictable effects on the availability of the monoamine neurotransmitters dopamine, noradrenaline, and serotonin. We hypothesized that 5HTTLPR genotype would show little association with prefrontal cognitive performance, but that COMT and MAOA would have interacting effects on cognition through their shared influence on prefrontal catecholamine availability. We assessed the individual and epistatic effects of functional polymorphisms in COMT, MAOA, and 5HTTLPR on children's prefrontal cognitive function in nearly 6,000 children from the population-based Avon Longitudinal Study of Parents and Children (ALSPAC). Neither MAOA nor 5HTTLPR polymorphisms showed significant effects on cognitive function. In boys but not girls, there was a modest but statistically significant interaction between MAOA and COMT genotypes such that increased prefrontal catecholamine availability was associated with better working memory. These results suggest that assessment of multiple genes within functionally related systems may improve our understanding of the genetic basis of cognition. © 2010 Wiley-Liss, Inc.Individual differences in cognitive function are highly heritable and most likely driven by multiple genes of small effect. Well-characterized common functional polymorphisms in the genes  Experimental manipulation of human and primate neurotransmission has provided insight into the distinct roles that the monoamine neurotransmitters serotonin (5-HT), dopamine (DA), and noradrenaline (NA) play in prefrontal cognition [Clarke et al., O-methyltransferase (COMT), one such enzyme [Karoum et al., COMT knockout mice show normal prefrontal NA levels and administration of the COMT-inhibitor tolcapone increases the release of DA, but not NA, in rat PFC [Gogos et al., Two major routes for the deactivation of monoamine neurotransmitters are reuptake by transporter molecules in the presynaptic cell membrane, and enzymatic degradation within the neuron or synapse. Both serotonin and NA transporters are abundant in PFC [Mantere et al., COMT, MAOA, and the serotonin transporter-linked polymorphic region (5HTTLPR) affect the level and/or activity of their products in defined manners, summarized in COMT Val allele was associated with worse performance on a measure of executive function, the Wisconsin Card Sort Test (WCST). While meta-analysis of similar studies shows little consistent cognitive effect [Barnett et al., MAOA and 5HTTLPR polymorphisms on cognition. MAOA genotype has been reported to significantly affect IQ among healthy Chinese women [Yu et al., 5HTTLPR polymorphism have been predominantly negative for standard neuropsychological measures such as IQ, response inhibition, and sustained attention [Fallgatter et al., Individual differences in cognitive function are highly heritable [Devlin et al., 158Met on cognitive performance at ages 8 and 10 in a population-based sample of more than 5,000 children from the Avon Longitudinal Study of Parents and Children (ALSPAC) [Barnett et al., MAOA and 5HTTLPR. We also assess evidence for epistasis between the MAOA, 5HTTLPR, and COMT polymorphisms, in an attempt to broaden the candidate gene approach to include assessment of functionally interactive gene systems, here represented by the overlapping effects of these SNPs on synaptic neurotransmitter availability. We hypothesized that genes that exclusively affect serotonergic function (5HTTLPR) would show little effect on these prefrontally driven cognitive tasks, but that main genetic effects, and potential epistasis, would be demonstrated for variants affecting dopaminergic or noradrenergic availability , with 5 µM of each primer and 25 mM dNTPs in a total reaction volume of 25 µl. Amplifications were performed on a Perkin Elmer 9700 thermocycler  with one cycle at 96°C for 10 min followed by 30 cycles of 94°C for 15 sec, 60°C for 15 sec, 72°C for 30 sec, and a final 3-min extension at 72°C. The forward primer was labeled with the fluorescent dye 6-FAM, and amplicons were visualized on an ABI 3100 capillary sequencer. Allele sizes  were determined using Genotyper 2.5 (Applied Biosystems). MAOA 3, 3.5, and 4-repeat allele frequencies in girls were consistent with Hardy–Weinberg equilibrium .DNA, obtained from blood and mouthwash samples, was extracted and processed [Jones et al., 5HTTLPR, low activity individuals were those with SS, SLG, or LGLG genotypes, medium activity were SLA or LGLA genotypes, and LALA genotypes were denoted high activity [Hu et al., COMT, Met/Met individuals were considered low, Val/Met individuals medium, and Val/Val individuals high activity, because Val allele carriers show higher COMT enzyme activity, and hence faster degradation of catecholamines at body temperature [Lachman et al., MAOA we excluded genotypes involving the rare 2- or 5-repeat alleles, whose functional effects are not yet clear [Sabol et al., MAOA is located on the X chromosome, boys carry only one allele. Boys with the 3-repeat allele were denoted low MAOA activity and those with 3.5- or 4-repeat alleles as high activity, since they show higher transcriptional efficiency and hence result in higher levels of synaptic MAOA [Sabol et al., Individuals were classified into high, medium, or low activity groups with respect to the functional effects of the polymorphism in each of the three genes. In each case, “high activity” groups comprised the alleles leading to fastest clearance of the synaptic neurotransmitter and “low activity” the alleles leading to slowest clearance. For 5HTTLPR, and for MAOA in girls only. For boys, we compared cognitive scores in high versus low activity groups using t-tests. Main effects of COMT in this sample have been previously reported [Barnett et al., Main effects of genes on cognition were assessed by comparing cognitive scores between high, medium, and low activity groups using one-way ANOVA for the 5HTTLPR and MAOA genotypes, and a catecholaminergic one between COMT and MAOA genotypes. We assessed evidence for these interactions using general linear models with each cognitive measure as an outcome variables predicted by two genetic main effects and a gene–gene interaction term. As before, all genotypes were coded as low, medium, or high activity. These analyses were conducted separately in boys and girls because MAOA is located on the X chromosome, and because COMT is known to have sexually dimorphic effects on a range of functional and neuropsychiatric phenotypes [Harrison and Tunbridge, Two possible sources of epistatic effects on cognition were considered: a serotonergic interaction between 5HTTLPR genotype. There were no cognitive differences between children for whom 5HTTLPR, MAOA, or COMT genotypes were or were not available . Allele frequencies were, for 5HTTLPR alleles S = 41.4%, LA = 51.5%, LG = 7.1%, and for MAOA, 2-repeat = 0.2%, 3-repeat = 34.0%, 3.5-repeat = 2.0%, 4-repeat = 62.4%, 5-repeat = 1.5%. COMT allele frequencies, as previously reported, were Val = 48.6%, Met = 51.4%. The number of children considered as low, medium, and high activity genotypes for whom at least one cognitive measure was available are shown in The number of children included in each analysis varied by the availability of cognitive data and genotypes: cognitive data were available on around two-thirds of those for whom DNA is available. As previously reported [Araya et al., MAOA on cognitive measures were assessed separately in boys and girls. t-Tests between high  and low (minimum n = 854) activity MAOA groups in boys showed no effects on any cognitive measure. Similarly in girls, one-way ANOVA between low, medium, and high activity MAOA groups  revealed no cognitive differences  such that better performance was seen in the high than medium activity group (Tukey's post hoc P = 0.03). One-way ANOVA revealed no cognitive differences between the three genotypes  within the low activity group.Groups defined by low, medium, and high serotonin transporter promoter activity were compared on all cognitive measures using one-way ANOVA. There were no differences between groups on measures of IQ or verbal inhibition see . NominalCOMT genotype and MAOA genotype, and a COMT × MAOA interaction term, were fitted separately for girls and boys. In boys, there was evidence of interaction between COMT and MAOA only for working memory score . Nonetheless, this sample remains the largest so far examined with respect to MAOA and cognition, and the complete lack of difference in cognitive score between genotypes in either sex is again a decisive negative result.Although the lack of cognitive effects of MAOA, there was a modest but significant interactive effect of MAOA genotype and COMT genotype on working memory performance in boys but not girls. With few exceptions, cognitive genetic studies have thus far had inadequately sized samples to realistically assess possible gene–gene (epistatic) interactions. The COMT × MAOA interaction observed here for working memory is plausible given the dependence of working memory on prefrontal dopaminergic availability. That no similar effect was found for IQ is surprising: COMT genotype has a strong, male-specific effect on verbal IQ in this sample [Barnett et al., MAOA on IQ but not measures of executive function [Yu et al., In contrast to the absence of main effects of Separate analyses were carried out for boys and girls and for multiple cognitive phenotypes. Since scores on the four cognitive measures were correlated, strict correction for multiple comparisons using, for example, a Bonferroni correction, was not suitable. Nonetheless, while the large sample used in the study reduces the chances of Type II error, the possibility of Type I error remains present.MAOA has a widespread distribution in the brain [Grimsby et al., MAOA expression may be predominantly determined by genetic and epigenetic factors other than this VNTR, including a currently unknown locus with which it shows strong linkage disequilibrium [Balciuniene et al., COMT expression appears greater in PFC than other regions [Matsumoto et al., COMT is complex, with tissue-specific monoallelic expression [Gimelbrant et al., COMT SNPs that act in concert with the Val158Met polymorphism in influencing cognitive function in this sample [Barnett et al., COMT haplotype groups produced no meaningful change in results from those seen using just the Val158Met genotype .While both tasks that have a high working memory content [Owen, MAOA genotype affects working memory only in the context of high prefrontal catecholamine availability. There is, of course, a distinction between demonstrating a statistical interaction, and understanding the biological mechanism of interaction between two genes [Clayton and McKeigue, COMT and/or MAOA expression affect cognitive function. One such possibility is NA, which, like DA, is catabolized to some extent by both COMT and MAOA. At present, dopaminergic pathways remain the more obvious candidate, because COMT appears to play a relatively minor role in regulating NA in the PFC [Gogos et al., The gene–gene interaction found here suggests that COMT × MAOA effect on working memory was in accordance with our previous study [Barnett et al., COMT genotype affected a wide range of cognitive functions in boys but had no effect in girls. There is additional wide-ranging evidence to suggest that the effects of COMT may be different between the sexes [Harrison and Tunbridge, COMT knockout mice, which show sex-specific behavioral phenotypes [Gogos et al., COMT in human PFC [Chen et al., COMT are the bilateral relationships between COMT and estrogen-related compounds: estrogens mediate COMT expression [Xie et al., COMT metabolizes catechol estrogens, a process regulated by Val158Met variation [Worda et al., COMT, the location of the MAOA gene on the X chromosome complicates association studies, with the result that little is known about the relative expression and effects of MAOA in the two sexes.The sex-specific nature of COMT and MAOA, two genes involved in the functional deactivation of DA and other neurotransmitters, show interactive effects on working memory performance in normal children from a large, homogeneous, and representative sample. While neither gene has been unquestionably linked to psychiatric disorder, normal variation in brain function is likely to be partly determined by genetic variation in neurotransmitter pathways. Understanding genetic effects on normal brain function is a necessary stage in understanding the basis of abnormal function, but large samples are required to tease out relatively subtle effects on cognitive development at the population level.In conclusion, we present evidence that functional polymorphisms in


Impulsivity can be defined as a predisposition toward rapid, unplanned reactions to internal or external stimuli regardless of negative consequences of these reactions for the impulsive individual or for others , lack of premeditation (difficulty in thinking and reflecting on consequences of an act), lack of perseverance (inability to remain focused on a task), and sensation seeking (tendency and openness to try and enjoy exciting or dangerous activities). The BIS-11 assesses impulsivity on the subscales attentional impulsivity (inability to focus attention or concentrate), motor impulsivity (acting without thinking), and non-planning impulsivity (lack of future orientation or forethought). Both questionnaires are highly correlated with each other (r = 0.67), but correlations between their subscales are only weak and inconsistent, supporting the notion that both measures cover different aspects of impulsivity (Meule et al., Beyond the fact that self-report and behavioral measures seem to capture different aspects of impulsivity, conceptualizations also vary between the different self-report instruments. For instance, two of the most widely used impulsivity questionnaires are the Only a few studies have done this as yet. For instance, differential relationships between subscales of the BIS-11 and eating disorder symptomatology have been found in clinical samples. Patients with binge eating disorder had higher scores on the motor impulsivity subscale compared to healthy controls, but did not differ on the other two subscales (Nasser et al., Eating Inventory (formerly Three-Factor Eating Questionnaire, Stunkard and Messick, hunger subscale and attentional impulsivity. Furthermore, both attentional and motor impulsivity were correlated with disinhibition (Lyke and Spinella, Studies investigating non-clinical samples also revealed differential associations between BIS-11 subscales and various measures of eating behavior. For example, Lyke and Spinella  examinedconcern for dieting subscale of the Restraint Scale (Herman and Mack, eating concern subscale of the Eating Disorder Examination—Questionnaire (Fairburn and Beglin, In a recent study by van Koningsbruggen and colleagues , only thr = 0.94, Spinella, While the BIS-11 contains 30 items, Spinella  developeFood Cravings Questionnaire—Trait, Cepeda-Benito et al., Mood Eating Scale, Jackson and Hawkins, Night Eating Questionnaire, Allison et al., Perceived Self-Regulatory Success in Dieting Scale, Meule et al., Restraint Scale, Herman and Mack, Yale Food Addiction Scale, Gearhardt et al., In a range of studies, positive correlations could be found between the BIS-15 and various constructs that are related to overeating (Table external eating subscale of the Dutch Eating Behavior Questionnaire (van Strien et al., Up to now, only one other research group has examined relations between eating behavior measures and BIS-15 subscales. In this study (Hou et al., In sum, it appears that attentional impulsivity is most consistently related to various measures that are associated with overeating. Positive, but less consistent, relationships can also be found with motor impulsivity, particularly in patients with binge eating behavior. Non-planning impulsivity seems to be only weakly related to overeating. Neither subscale seems to be consistently correlated with BMI, which may be due to the fact that BMI is influenced by many factors other than eating behavior. Future research needs to address the question why attentional impulsivity in particular has such a prominent role in overeating. First evidence suggests that attentional impulsivity may increase the susceptibility that highly palatable food-cues attract attention and trigger eating behavior (Hou et al.,


Evidence-based medicine is supposed to be a conclusive summary of available empirical knowledge on certain medical issues and as such serving as the basis of guidelines and treatment recommendations. Meant to be self-explaining, the term actually appears to be crucial. It seems inconsistent and inconclusive since recommendations of single countries/institutions are based on different evidence measures—some derive evidence from meta-analyses, whereas others exclusively follow randomised controlled studies. In a philosophical side trip, Möller  criticalIn mild major, minor or subsyndromal depression, the total score of the Hamilton Depression Rating Scale (HAMD) so far is regarded as ‘gold standard’ in evaluating efficacy of antidepressant treatment. Nevertheless, in comparing the full scales HAMD17 and the Inventory of Depressive Symptomatology IDS-C28) plus diverse unidimensional subcales in a randomised, placebo-controlled trial in a representative patient sample, Helmreich et al. [8 plus diGRIA1, GRIA2 and GRIA4, Chiesa et al. [In the pathophysiology of depression, glutamate plays an important role. Exploring the association of major depressive disorder with some single nucleotide polymorphisms within the glutamatergic AMPA receptor subunits a et al.  in a relAlthough an association between affective disorders and the metabolic syndrome has been suspected, past studies did not bear clear results in this respect. Hence, Kahl et al.  have exaCognitive deficits play a major role in psychiatric disorders like depression and schizophrenia, but also in neurodegenerative diseases. In an fMRI study in healthy probands, Voss et al.  investigDTI also was used by Konrad et al.  to invesIn all, cognitive deficits are core symptoms of psychiatric disorders and responsible for an unfavourable outcome. The investigation of the neurobiological basis of these symptoms is needed to develop new regenerative treatment strategies in psychiatric and neurodegenerative diseases.


A commentary onNeural correlates of moral sensitivity in obsessive compulsive disorderby Harrison, B. J., Pujol, J., Soriano-Mas, C., Hernández-Ribas, R., López-Solà, M., Ortiz, H., et al. (2012). Arch. Gen. Psychiatry 69, 741–749.Archives of General Psychiatry, Harrison et al. (In a recent issue of n et al.  providedThese brain areas were found to be active on healthy humans experiencing moral emotions such as indignation or moral disgust (see Moll et al., The results are striking in so far as they show that patients with OCD demonstrated significantly increased activation of the ventral frontal cortex, particularly of the medial orbitofrontal cortex. Moreover, significant positive associations were documented between the patients' DY-BOCS ratings of total symptom severity and the activation of the anterior insula (Harrison et al., The evidence from this study offers the opportunity to discuss two emerging related issues.First, it adds further support on a clinical model to the view that visceral and moral disgust share, at least in part, common neural and cognitive mechanisms Jones, . In factThus, their heightened moral sensitivity might reflect an enhanced disgust for immoral outcomes.This hypothesis is supported by previous evidence coming from neuroimaging studies. For instance, an overlapped recruitment of ventromedial prefrontal and orbitofrontal cortices have already been reported during the processing of both sensorial and moral disgust (Moll et al., A similar result was announced for the insula. For example, Wicker et al.  have disAll these findings suggest that, the heightened moral sensitivity in OCD can be grounded on the same neural mechanisms responsible for their altered sensitivity to outcomes which can induce sensory and emotional disgust (Shapira et al., Another remarkable aspect deserving some discussion is that OCD participants were not different from healthy control subjects in the subjective rating of the moral dilemmas, despite the difference reported in neural activation. This result, which probably argues some distinction between the patients' increased neural responses and their perceived emotional experience during a moral dilemma (Harrison et al., This suggestion is supported by the recent evidence of a negative correlation between interoception and anxiety (Pollatos et al.,


To the Editor:In our manuscript entitled “Single-neuron RNA-Seq: technical feasibility and reproducibility,” published online on July 6th, 2012, we have made a few serious mistakes which need to be corrected:J. Neurosci. 31, 6939–6943.”We referenced several points and quoted statements from a recent study by Okaty et al., but we neither cited the paper nor placed quotation marks on the statements. The full citation should be “Okaty, B. W., Sugino, K., and Nelson, S. B. (2011). Cell type-specific transcriptomics in the brain. We request to add a citation after “The analysis of each single-cell transcriptome consists of several independent steps” in the first paragraph of Results, as “The analysis of each single-cell transcriptome consists of several independent steps ,” given that these points were previously raised by Okaty et al. We request to add a citation to the first paragraph of Introduction, as “studies of gene expression and function in the brain were restricted to a relatively small number of genes (Luo and Geschwind, Moreover, gene expression may be regulated in opposing directions in different cell types, thereby appearing static in composite data.ToMoreover, as previously discussed by Okaty et al., “gene expression may be regulated in opposing directions in different cell types, thereby appearing static in composite data (Okaty et al., We apologize to the readers of Frontiers in Genetics and to the authors of the Okaty et al., manuscript, for our failure to correct these mistakes during the review process of our manuscript.


Frontiers in Pharmacology brings together a group of review and original articles focusing on IBS.Irritable bowel syndrome (IBS) is a very common functional disorder of the digestive tract. Despite the prevalence and the social impact of IBS, the exact etiopathogenesis is incomplete and the pharmacological treatment is unsatisfactory. In this research topic, Fabrizio De Ponti brilliantly outlined the drugs currently used in—or in development for—IBS, within a scenario in which the specific armamentarium of medications is unsatisfactory  activation on the production of kynurenine  in IBS  and selective  5-HT4 receptor agonist in vitro with strong in vivo gastrointestinal activity in guinea pigs, rats, dogs, and in healthy humans. TD-8954 may have value in the treatment in C-IBS sufferers.Beattie et al. investigated the Interleukin-6 (IL-6) is elevated in the plasma of D-IBS patients (Rana et al., 1 and/or CB2 receptor activation—exert pharmacological actions which are potentially beneficial in IBS (Izzo and Coutts, 1 and CB2 receptor agonists in normalizing the accelerated transit (Kimball et al., 1 and CB2 responsiveness is maintained long after an initial inflammatory period and suggest a role of cannabinoid receptors in the underlying pathophysiology of PI-IBS.Cannabinoids—via CBThe differentiated Caco-2 cells intestinal cell line, derived from a human colon carcinoma has been used for drug absorption studies as well as to evaluate the epithelial barrier integrity in relation to IBS as well as to investigate the intestinal permeability of IBS drugs (Catalioto et al., In summary, this research topic should provide a useful resource for IBS researchers, both basic and clinical. The pharmacological studies, together with new strategies for drug discovery, highlight that more research is urgently required to translate novel and innovative basic concepts into prescribing options for healthcare professionals. It is hoped that this collection of articles will inspire further research into IBS.


The fundamental role of D-serine as a co-agonist at the N-methyl-D-aspartate receptor (NMDAR), mediating both physiological actions of glutamate in long term potentiation and nociception and also pathological effects mediated by excitotoxicty, are well-established. More recently, a direct link to a chronic neurodegenerative disease, amyotrophic lateral sclerosis/motor neuron disease (ALS) has been suggested by findings that D-serine levels are elevated in sporadic ALS and the G93A SOD1 model of ALS (Sasabe et al.,  It has only recently been recognized that D-amino acids play an important physiological role in the central nervous system. The evidence is most convincingly demonstrated for D-serine through its action as a co-agonist at the N-methyl-D-aspartate receptor (NMDAR), a major glutamate receptor subtype, involved in synaptic plasticity , TAR DNA binding protein 43 (TARDBP) and fused in sarcoma (FUS). Arginine199 is highly conserved across species from bacteria to man and lies within the active site close to the FAD and substrate binding sites of the enzyme. The effect of the R199W-DAO mutation is very profound abolishing enzyme activity , the major biosynthetic for D-serine, in mammalian brain has been characterized extensively  was backcrossed with C57BL/6J mice and maintained as homozygotes (DAO−/−) in order to study the motor phenotype in more detail  has been identified in a mouse line, which shows abnormal locomotor activity  and in two overlapping ALS-like conditions, progressive bulbar palsy and primary lateral sclerosis , compared to normal controls and a control neurological group consisting of conditions associated with similar muscle loss   compared to controls, the greatest change being an increase of 57% in glycine which suggested a potential impairment in glycine transport (de Belleroche et al., An impaired clearance of both glycine, as indicated from these results (Lane et al., −/−, where D-serine synthesis is reduced. On the other hand, glycine facilitates NMDAR-mediated currents after afferent stimulation, diminished by GO but not by DAO (Li et al., −/− knockdown or depletion of glycine by GO reduce LTP (Li et al., In hippocampus and amygdala (Li et al., The interplay between glycine site co-agonists has not been extensively characterized in brain stem and spinal cord. However, studies in rat hypoglossal motor neurons suggest that an increase in synaptically released glycine in response to metabolic stress may play a role in exacerbating NMDA receptor-mediated excitotoxicity in motor neurons (Kono et al., Experimental evidence obtained to date highlights the selective localization of DAO in spinal cord motor neurons, brain stem motor nuclei, cerebellum and other CNS centers involved in motor control. DAO is also found in glial cells in spinal cord and cerebellum. This distribution of enzyme activity is consistent with the lower levels of D-serine found in posterior brain compared to forebrain. The primary findings that implicate D-serine in ALS and potentially in neurodegeneration in general are that elevated levels of D-serine in spinal cord are found in ALS and the SOD1 mouse model of ALS (Sasabe et al., In order to further substantiate this concept, we have explored the potential toxic properties of the FALS-associated R199W-DAO mutation in cell culture and have shown that whether expressed in astrocytes or motor neurons the mutation promotes apoptosis but most interesting when expressed in glial cells the mutant protein is able to initiate apoptosis in neighboring neurons in co-culture that lack the mutation. The way in which this effect is transmitted between cells appears to be crucially dependent on the presence of an NMDAR co-agonist as the effect is blocked by the selective antagonist DCKA. In the situation where DAO is inhibited due to the presence of a mutation, D-serine levels are elevated (Sasabe et al., It is of considerable importance to note that increased synthesis of D-serine by neuronal and glial SR is known to be activated by cellular stressors such as AMPA, amyloid-beta and lipopolysaccharide (Wu and Barger, The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Dear Sir,Upsala Journal of Medical Sciences. This article provides an interesting analysis of estimating renal function in critically ill patients, but I believe there are several aspects that may warrant further discussion:I read with great interest the article entitled ‘Significant differences when using creatinine, modification of diet in renal disease, or cystatin C for estimating glomerular filtration rate in ICU patients’ by Lipcsey et al.  in the MFirst of all, the equations estimating glomerular filtration rate (GFR) are all notoriously flawed in the setting of rapidly changing renal function, that is, acute kidney injury. The creatinine-based formula for creatinine clearance is shown to over-estimate renal function when actual GFR is low, due to its multiple interfering factors and tubular secretion nature, but the cystatin C-based formula is also affected by muscle mass and adiposity . To compSecond, the duration of illnesses and timing of tests are also not taken into consideration in Lipcsey's article. Nejat et al. in their multicenter observation study have indicated that cystatin C rises earlier than creatinine in patients with acute kidney injury by 4–5 hours from a general ICU population .Finally, is there any better way of determining renal function in the critically ill with convenient point-of-care feasibility? Tidman et al. in their study of GFR determination in chronic kidney disease patients have provided a new thought: combining creatinine-based and cystatin C-based results for estimation . Though