Ovis orientalis), 22 wild goat (Capra aegagrus), nine Indian gazelle (Gazella bennettii) and eight Goitered gazelle (Gazella subgutturosa) during the national survey for wildlife diseases in Iran. Serum samples were evaluated using serologic antibody tests for Peste de petits ruminants virus (PPRV), Pestiviruses [Border Disease virus (BVD) and Bovine Viral Diarrhoea virus (BVDV)], Bluetongue virus (BTV), Bovine herpesvirus type 1 (BHV-1), and Parainfluenza type 3 (PI3). Sera were also ELISA tested for Pestivirus antigen. Tissue samples including spleen, liver, lung, tonsils, mesenteric and mediastinal lymph nodes and white blood cells (WBCs) were tested using polymerase chain reaction (PCR) for PPRV, Foot and Mouth Disease virus (FMDV), Pestivirus, BTV, Ovine herpesvirus type 2 (OvHV-2) and BHV-1. Serologic tests were positive for antibodies against PPRV (17%), Pestiviruses (2%) and BTV (2%). No antibodies were detected for BHV-1 or PI3, and no Pestivirus antigen was detected. PCR results were positive for PPRV (7.8%), FMDV (11%), BTV (3%), OvHV-2 (31%) and BHV-1 (1.5%). None of the samples were positive for Pestiviruses.A molecular and serological survey of selected viruses in free-ranging wild ruminants was conducted in 13 different districts in Iran. Samples were collected from 64 small wild ruminants belonging to four different species including 25 Mouflon ( Habitat fragmentation, hunting, and infectious diseases often threaten biodiversity and may contribute to significant declines in wildlife populations , 2. ThesWide climatic variations and untouched landscapes in Iran support a diverse mammalian fauna including 191 species from 93 genera and ten orders , 11. WilMost of the wild ruminants in Iran are on the International Union for the Conservation of Nature (IUCN) Red List as a direct result of infectious diseases, illegal hunting and drought .Goitered gazelle, listed by the International Union for Conservation of Nature (IUCN) red list as vulnerable, are found in the Zargros region of Iran. Indian gazelle, protected in Iran but a considerable population of Indian gazelle are found in the Kavir National Park . Both WiMorbillivirus (family Paramyxoviridae) closely related to the eradicated rinderpest virus, causes the disease Peste des petits ruminants (PPR) which is considered to be one of the most significant livestock diseases in the Middle East, Africa and Asia [The majority of pathogens that cause disease outbreaks in wild ruminants worldwide are viral . Peste dand Asia , 17. PPRand Asia . Multipland Asia , 20. PPRand Asia , 22.Foot and Mouth Disease (FMD) is caused by a single-stranded negative sense RNA virus that belongs to the genus Aphthovirus from the Picornaviridae family. The FMD virus (FMDV) contains seven different serotypes, O, A, C, Asia 1, Southern African Territories (SAT) 1, SAT 2 and SAT 3 . MoleculPestivirus infections in animals are caused by a group of viruses from the Flaviviridae family, genus pestivirus. Three different pestiviruses have been described, namely, bovine viral diarrhoea virus (BVDV), Border disease virus (BDV) and Classical Swine Fever Virus (CSFV) . BVDV anOrbivirus from the family Reoviridae. There are 24 antigenically distinct serotypes in the genus and differing levels of cross reactivity have been observed between different serotypes[Culicoides midges [Another important infection seen in domestic and wild ruminants is Bluetongue virus (BTV) an serotypes. Epidemiserotypes. Sero-epserotypes–44. Howeserotypes. BTV cirs midges , 46 withs midges .Malignant Catarrhal Fever (MCF) is caused by members of subfamily Gammaherpesvirinae, genus Macavirus . AlcelapInfectious bovine rhinotracheitis virus (IBRV) and parainfluenza type 3 virus (PI3V) are the main viral respiratory diseases in livestock worldwide , 57.Paramyxovirus family, is one of the most common viral respiratory infections in cattle. Although PI3V can cause primary respiratory disease in cattle, it usually leads to the development of secondary bacterial infections in the respiratory tract of large ruminants [Infectious bovine rhinotracheitis (IBRV), caused by Bovine herpesvirus 1 (BHV-1), is associated with several clinical features in cattle. BHV-1 can produce a lifelong latent infection with occasional reactivation of the virus followed by viral shedding for several days. A serological positive response in animals is evidence of a latent infection . Parainfuminants . Both seuminants .The exteuminants .The aim of this study was to investigate if there is evidence of these commonly seen livestock viruses in wild ruminants in the national parks and protected regions of Iran using serological and molecular techniques.Ovis orientalis), 22 wild goat (Capra aegagrus), nine Indian gazelle (Gazella bennettii) and eight Goitered gazelle (Gazella subgutturosa).The study animals comprised wild ruminants from four different species including 25 Mouflon   each indWhole blood was collected from the jugular vein, by cardiac aspiration or from the thoracic or abdominal cavities during fresh field necropsies by an authorized veterinarian from the Iran DOE. The collected blood was divided into two tubes, with and without anti-coagulant (EDTA) and labelled. Samples were transported, refrigerated, to the Virology laboratory at the Faculty of Veterinary Medicine at the University of Tehran within 24 hours.The names of the regions and their geographic information system coordinates for all sampled animals and the age and sex of wildlife species tested in this study are shown in Tables At the laboratory, serum was separated by centrifugation at 2000 RPM for 10 min. White blood cells from each blood sample was separated for antigen detection and RNA/ DNA isolation for molecular detection.WBCs were isolated using Ficoll-Hypaque density gradient separation method. In brief, five mL of blood was diluted two-fold in sterile phosphate buffered saline (PBS) then added into the 10 mL of Ficoll-Hypaque (Sigma Aldrich), centrifuged for 400 × g at room temperature for 30 min. The cells were isolated and washed twice with PBS . AliquotThe study was approved by the Research and Ethics Committee of Iran Department of Environment. Animals were shot by authorized hunters and autopsied by veterinarians who specialized in wildlife studies during the national programme for survey of wildlife diseases. The hunting licences were issued by the Iran Department of Environment for the DOE qualified hunters.Serological tests used to detect antibodies included the following: a) PPR cELISA kit , b) pestivirus blocking ELISA kit (BVDV) and (BDV) , c) Bluetongue competitive ELISA  and d) Bovine Herpesvirus 1 (BHV-1) ELISA kit and e) virus neutralization test (VNT) for parainfluenza type 3 . PestiviIn all serological ELISA tests positive and negative control sera were included in each plate. In addition to the control samples (provided within the ELISA kits), two more known positive and negative samples were included in each round of ELISA or VNT tests. The positive and negative-control serum samples were serum from animals who had been tested at the diagnostic virology unit at the Razi Vaccine and Serum Research Institute or Virology laboratory at the Faculty of Veterinary Medicine at the University of Tehran, Iran and kept as standard control samples. Each of these serum samples were previously evaluated in different assays. For virus neutralization test, each serum sample was tested in two independent assay runs and serum samples with a titre of 1/64 or higher were considered positive . All of White blood cells and the fresh/frozen tissues from all animals were subject to DNA and RNA isolation. RNA extraction was performed using QIAamp Viral RNA Mini kit  according to the manufacturer's instructions. The cDNA was synthesized from extracted RNA using Moloney Murine Leukemia Virus-Reverse Transcriptase (MMLV-RT) and random primers . Total DNA was isolated from the above mentioned samples using QIAamp DNA Mini Kit  according to the manufacturer's instructions. The synthesized cDNA and isolated DNA were saved as aliquots and stored at −80°C for further analysis.Previously published PCR protocols were used to detect; a) PPR , b) FMD Serological exposure of the 64 wild ruminants to PPR, BVD and BD, BTV, BHV-1, PI3 and pestivirus antigen is shown in The PCR results for PPRV, FMDV, Pestivirus, BTV, OvHV-2 and BHV-1 are shown in Four mouflon, two wild goats and one Indian gazelle were positive for FMDV by PCR on tonsil samples. The only positive Indian gazelle for FMDV was from Tooran and the positive Mouflon were from Tooran (n = 1), Khojir (n = 2) and Angooran (n = 1). Two FMDV positive wild goats were from Khojir National park and Mujen protected region. All positive PCR tests for PPRV, FMDV, BHV-1 and four of the OvHV-2 samples were confirmed by sequencing. Edited nucleotide sequences were compared to the nucleotide sequence database for similarity using BLASTn search tool and was shown to have significant homology to the available sequences for the listed viruses. –S5 FilesThis is the first survey of viral diseases in wild ruminants in Iran. It will provide valuable information for future research in disease ecology and risks associated with transmission of these viruses between wildlife and livestock.Studies have shown that PPRV is endemic in Iran and the other neighbouring countries with multiple devastating outbreaks occurring in domestic and wild ruminants , 22. MorBorder disease is considered as an endemic disease in domestic sheep and goats in Iran  and so MFoot and mouth disease has been reported in Mouflon, wild goats and Indian gazelles in several countries , 35. As Previous work in southeast Iran identified the serological existence of Bluetongue virus in domestic sheep with no reports of clinical signs , 44, 78.In this survey all of the seropositive and PCR positive BT samples were seen in Mouflon from Bamo National Park. The locality of this survey overlaps geographically with the district where a high prevalence of BTV is seen in livestock .There was no serological evidence for PI3 and IBR, the main viral infections of Iranian livestock, and there was just one Indian gazelle positive for BHV 1 using PCR. These viruses mainly cause disease in intensively managed livestock populations, so the lack of seroprevalence was expected in wildlife. In addition, BHV 1 infection is rarely seen in small ruminants and the clinical form is mainly associated with intensive feedlot or dairy production industries .PCR tests showed a high prevalence of OvHV-2 in Mouflon. The virus does not produce clinical signs in small ruminants and there have been no reports of clinical disease in wild and domestic sheep and goats , 81. In In conclusion, while there is clear evidence that transboundary disease viruses particularly FMDV, BTV and PPRV are circulating in wild ruminants. Further studies are required to investigate the transmission pathways and disease ecology between wildlife and livestock.S1 FileGenBank accession numbers are shown at the left side of the figure and Iranian isolates are identified with double asterisk marks.(PDF)Click here for additional data file.S2 FileGenBank accession numbers are shown at the left side of the figure and Iranian isolates are identified with double asterisk marks.(PDF)Click here for additional data file.S3 FileGenBank accession numbers are shown at the left side of the figure and Iranian isolates are identified with double asterisk marks.(PDF)Click here for additional data file.S4 FileGenBank accession numbers are shown at the left side of the figure and Iranian isolates are identified with double asterisk marks.(PDF)Click here for additional data file.S5 FileGenBank accession numbers are shown at the left side of the figure and Iranian isolates are identified with double asterisk marks.(PDF)Click here for additional data file.


SH2 domain-containing tyrosine phosphatase-2 (PTPN11 or Shp2) is a ubiquitously expressed protein that plays a key regulatory role in cell proliferation, differentiation and growth factor (GF) signaling. This enzyme is well expressed in various retinal neurons and has emerged as an important player in regulating survival signaling networks in the neuronal tissues. The non-receptor phosphatase can translocate to lipid rafts in the membrane and has been implicated to regulate several signaling modules including PI3K/Akt, JAK-STAT and Mitogen Activated Protein Kinase (MAPK) pathways in a wide range of biochemical processes in healthy and diseased states. This review focuses on the roles of Shp2 phosphatase in regulating brain-derived neurotrophic factor (BDNF) neurotrophin signaling pathways and discusses its cross-talk with various GF and downstream signaling pathways in the retina. PTPN11 gene  is a 593 amino acid non-transmembrane protein tyrosine phosphatase (PTP) encoded by Shp2 plays prominent biological roles in regulating several signal transduction cascades affiliated with its function in the early development of vertebrates, cell proliferation, differentiation, transcription regulation and metabolic control  as well as multiple GF signaling networks by the Shp2 phosphatase and its implications in the retina.In the retina, Shp2 is well expressed in the ganglion cell layer (GCL) and inner nuclear layer (INL) and its reactivity has been detected in photoreceptors  are secreted proteins that regulate neural growth, survival and function by negatively affecting the induction of various cellular apoptotic pathways. Retinal ganglion cells (RGCs) express various neurotrophic factors and are also supported by neurotrophic factors obtained locally from the Muller cells and retrogradely from the brain through axonal flow , the prominent structural constituent of caveolae Figure . Shp2 waThe role of other significant caveolar proteins such as Cavin family members particularly polymerase I and transcript release factor (PTRF) which participate in caveolae formation through its interaction with Cav-1, have not been extensively investigated and their potential cross-talk with Shp2 remains to be explored  and in the RGCs In cerebellar granule neurons, Shp2 suppressing functions are reported to abolish axonal regeneration by paired immunoglobulin-like receptor PIR-B/Shp mediated TrkB inhibition , another member of the neurotrophin family, exerts its survival-promoting effects by stimulating neural development and differentiation through MAPK cascade  signaling in human glial cells in response to EGF treatment. Zhang et al.  demonstrThe EGF-inhibitory impact of Shp2 was also investigated in a glioma cell line (SNB19). Despite lack of proliferation ability of SNB19 in response to EGF stimulation, it was found that interfering Shp2 mutant could reverse the cell’s ability to proliferate following EGF induction  signaling cascade which has major functions in cellular processes such as proliferation, differentiation and apoptosis  downstream signaling including Ras, Raf, MEK and MAPK cascade. Insulin receptor substrate 1 (IRS-1), one of the major IR substrate is a multisite docking protein which interacts with SH2 domain-containing proteins such as Shp2. Milarski and Saltiel  showed tPTPN11 gene mutation (Tartaglia et al., PTPN11 gene in animal studies resulted in extensive retinal degeneration, cell death and optic neuropathy during various developmental stages highlighting its regulatory function in retinal neuroprotection and progenitor retinal cell fate (Cai et al., Shp2 involvement in regulating retinal survival signaling pathways links dysregulation of this phosphatase to various physiological and pathological conditions. Under glaucomatous stress Shp2 leads to preferential RGC degeneration by inhibiting BDNF/TrkB downstream signaling through dephosphorylation and deactivation of TrkB (Gupta et al., in vivo and in vitro studies implied that the major effect of the phosphatase is mediated through its regulatory effects on various GFs and their downstream effectors which activates multiple signaling pathways. In this review we outlined and analyzed the existing evidence regarding Shp2 involvement in BDNF and other GF-dependant signaling networks with a specific focus on the retinal neuronal cells.Shp2 plays an emerging and important role in the retinal development and its preservation. Preliminary Although significant breakthroughs in the functional characterization of Shp2 have provided more knowledge of the physiological importance of the phosphatase, other areas of future study including manipulating Shp2 expression in retinal and other neuronal cell lineage or development of specific phosphatase inhibitors will further define the mechanisms through which Shp2 positively or negatively mediates various retinal signaling pathways. Another fascinating aspect would be considering Shp2 as a potential molecular target to modulate other neuronal signaling pathways thereby serving as mechanism-based therapy. The involvement of Shp2 in broad range of biochemical actions may make it a challenge to develop any specific therapeutic strategy to isolate a particular neuroprotective role. However, prospective investigations probing Shp2 protein expression, post-translational modifications, sub-cellular localization and interactome changes will unravel the role of this protein in various neurodegenerative disease and retinal disorders.The review was conceptualized, written and edited by each of the authors. Supervisor: SLG.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.


To the Editor: Zika virus infection is an emerging arboviral disease linked to an increased risk for severe neurologic outcomes and devastating adverse fetal complications, including pregnancy loss and congenital microcephaly (A 51-year-old man (patient 1) who regularly traveled to Samoa, a Pacific island with known Zika virus transmission, returned to New Zealand in late January 2016. He reported having been sexually abstinent while overseas. After 1 week he experienced onset of fever, rash, arthralgia, and ankle edema. Aware of the symptoms of mosquito-borne illnesses prevalent in his region of travel, he assumed he had a mild case of chikungunya and sought no medical attention.A 53-year-old woman (patient 2) who was the sexual partner of patient 1 experienced onset of sore throat, fever, arthralgia, and rash 17 days after her partner’s return to New Zealand (10 days after he had onset of symptoms). She had had no recent overseas travel, had no blood transfusions, and was unaware of having any mosquito bites. Four days after symptom onset, she visited a general practitioner, whose investigations revealed mild neutropenia, normal liver and renal function, no evidence of streptococcal throat infection, and a demonstrated immunity to measles.The rash subsequently spread, and small joint arthralgia worsened. The patient returned to the general practitioner 9 days after symptom onset. Disclosing her partner’s travel history and symptoms and reporting having unprotected sexual intercourse with him after his return to New Zealand (before and during the time he was symptomatic), she enquired about the possibility of Zika virus infection. After discussing the matter with a clinical microbiologist and securing the consent of the woman and her partner, the general practitioner ordered tests for arboviruses common in the male partner’s region of travel. All laboratory tests for chikungunya and dengue viruses were negative for both patients. Testing for Zika virus RNA was performed by using real-time reverse transcription PCR (rRT-PCR) in accordance with published methods  (9). Sert] 25). Semen collected on days 35 and 76 also tested positive . Semen samples collected on days 99 and 117 tested negative for Zika virus RNA. Attempts at virus isolation from the semen sample collected on day 23 failed to cultivate infectious particles.Laboratory investigations for Zika virus RNA in patient 1 demonstrated negative rRT-PCR results for serum (first tested 19 days after symptom onset) and for urine (first tested 21 days after symptom onset). Serologic testing performed on day 21 detected Zika IgM and IgG (IgG titer 1:320). Semen collected on day 23 tested positive for Zika virus RNA (cycle threshold [CAedes species of mosquito capable of transmitting Zika virus infection is established in the country (It is very unlikely that transmission of Zika virus infection to patient 2 occurred through a mosquito bite. Although occasional interceptions of exotic mosquito species have occurred at international ports of entry into New Zealand, neither of the


The purpose of this study was twofold: (1) to compare whether children from polygamous family structures significantly differ from children from monogamous family structures with regard to the frequency of parent-child conflict, and (2) whether children from these two structures employ different patterns of family conflict resolution.To address these questions, a random sample of 212 high school students (60.8% monogamous) completed a self-administered survey. The results of MANOVA show no significant differences (p > 0.05) between these two structures with regard to the frequency of parent-child conflict. The results also show similar conflict management styles between these two family structures within each of the following five domains .This study is unique in that it is the first empirical research to be conducted in the field of conflict resolution among youth and adolescents in polygamous marital structures and therefore, further investigation is needed to replicate these results utilizing different cross-cultural populations practicing polygamy.


A key factor in determining the likely outcome for a patient with colorectal cancer is whether or not the tumour has metastasised to the lymph nodes—information which is also important in assessing any possibilities of lymph node resection so as to improve survival. In this review we perform a wide-range assessment of literature relating to recent developments in gene expression profiling (GEP) of the primary tumour, to determine their utility in assessing node status. A set of characteristic genes seems to be involved in the prediction of lymph node metastasis (LNM) in colorectal patients. Hence, GEP is applicable in personalised/individualised/tailored therapies and provides insights into developing novel therapeutic targets. Not only is GEP useful in prediction of LNM, but it also allows classification based on differences such as sample size, target gene expression, and examination method. Colorectal cancer (CRC) is one of the most common causes of cancer-related deaths worldwide . Currenthttps://www.verywell.com, Updated 9 April 2017). In another report, nearly 30% of CRC cases with no record of their lymph node metastasis (LNM) status were found during the five-year follow-up to have died because of tumour recurrence. The authors of the latter study concluded that the presence of the occult lymph node metastases (micrometastases) that were not detected by routine examinations might be the cause [Curative surgery improves the CRC patient’s status at least in early stage, but the healing process is not adequate by itself . For exahe cause .The prevalence of micrometastases in patients with early CRC has recently been widely investigated. Published data indicate that even patients with stage I, who seem to have localised disease, may harbor micrometastases that were missed during the clinicopathological tests . In line(1)(a)chromosomal instability (CIN) ,(b)CpG island methylator phenotype (CIMP) ,(c)microsatellite instability (MSI) .A model that consists of three molecular subtypes including:(2)(a)CMS1 (14%), microsatellite instability (MSI) and immune hyperactivation,(b)CMS2 (37%), epithelial involvement, wingless-type MMTV integration site family member (WNT) and MYC pathway interaction,(c)CMS3 (13%), epithelial and metabolic involvement,(d)CMS4 (23%), invasive and metastatic activation of transforming growth factor–β (TGF-β) .A model of four consensus molecular subtypes (CMSs). The members of this model show the following discriminating features:Like other cancer types, CRC occurs by multiple misregulation of oncoproteins or tumour suppressors that impair the intra- and extra-cellular signal balances. The two most applicable models in this field are as follows:These models facilitate the identification of germline mutations of the genes involved in DNA mismatch repair (MMR) including MutS protein homolog 2 and 6 , MutL protein homolog 1 (MLH1), and PMS1 homolog 2 (PMS2), which all cause MSI in patients with hereditary HNPCC/Lynch syndrome . As in MAs previously mentioned, DNA mutations have a huge impact on prognosis and survival rates. However, the impact of thousands of detected mutations on cancer progression has not yet been elucidated, and we are not sure whether some of these mutations cause the cancer (driver mutations) or emerge because of the cancer (passenger mutations). Currently, gene expression profiling of the primary tumours has received a great deal of attention due to its ability to create a detailed picture of the genetic and epigenetic alteration status of tumours.What is the actual value of the gene expression signatures for the prediction of key events such as recurrence or LNM in CRC patients? This question has been the subject of numerous previous studies . GeneralThe detection of high-risk patients with recurrent CRC is arguably the main challenge. In this regard, applying molecular assessment tools could be helpful in identifying cases of occult micrometastasis and potential suitability of adjuvant therapy ,41. AccoOver the last decade, gene expression profiling by microarray has been a pioneering method in the detection of disease-related molecules. Compared with other standard techniques such as RT-PCR, which is not able to evaluate a large number of targets at once, microarray screening provides fast and reliable data with high accuracy from various samples. With microarray, it is now possible to analyse the whole expression pattern of a human genome within 48–72 h, thus gathering precious details about related molecular subtypes .Using cDNA microarray technology, Kwon et al. (2004) examined and compared the expression profiling of 4608 genes in 12 CRC tumours versus in noncancerous tissues . AccordiWang et al. (2004) tried to identify new prognostic markers for cancer relapse by DNA chip technology. Their achievement was a 23-gene panel allowing prediction of recurrence in Dukes’ B patients. The panel also provided insight regarding the underlying biological mechanism of rapid metastases; some of these genes are involved with tumour development and cell proliferation. For example, Tyrosine 3 mono-oxygenase tryptophan 5-monooxygenase activation proteins (YWHAH) and Regulator of chromosome condensation 1 (RCC1) are the most important genes governing the G2 checkpoint of the cell cycle and chromosome condensation initiation, respectively . In the Arango et al. (2005) proved that the downregulation of Ras homolog gene, a small GTPase protein known to regulate the actin cytoskeleton, was correlated with shorter survival. This approach is not only useful in Dukes’ C patients’ recurrence predictions but also can be used in the design of clinical management algorithms .Meeh and colleagues used digital long serial analysis of gene expression to elucidate the differences between node-negative and node-positive colorectal tumours. They reported that the development of node-positive CRC occurs, in part, through elevated levels of epithelial Fibronectin 1 (FN1). They suggested that the progression of the CRC from node-negative to node-positive disease may be facilitated partially by FN1 deregulation and the subsequent enhancement of tumour cell migration .In 2009, Watanabe and colleagues determined that gene expression programming could be a useful tool in predicting recurrence in stage III colorectal cancer, and also identified calcineurin binding protein 1 (CABIN1) among discriminating genes that may play a key role in the development of recurrence . AdditioThe other family involved in cancer metastasis is the Forkhead box proteins (FOX). For example, FOXC2, also known as Forkhead Box C2 (FKHL14), has been found to be significantly elevated in patients with lymph node involvement and correlated with the degree of LNM . UpregulIn 2011, Salazar and colleagues developed a robust gene expression classifier (ColoPrint) that can predict relapses in patients with early-stage CRC. According to their study, this method can identify patients with stage II CRC who will experience a recurrence within five years after surgery . The tecBesides ColoPrint, Lenehan et al. (2012) have developed a molecular prognostic examination able to identify tumour recurrence within three years in CRC cases having curative surgery. From analysing the expression changes of 18 key genes involved in regulation of cell signal transduction, gene expression, invasion, growth, angiogenesis, apoptosis, and antioxidation, they identified five genes that could be used in the prediction of tumour recurrence in CRC patients: BMI-1 polycomb ring finger oncogene (BMI), Vascular endothelial growth factor A (VEGFA), Ribosomal protein S10 (RPS10), Ets variant 6 (ETV6), and H3 histone, family 3B (H3F3B). However, the test was validated in stage I and II patients and the authors concluded that ≥12 lymph node samples would be required for accurate prognostication .The results of another study on 196 genes in CRC patients determined that use of the expression pattern of main genes such as Annexin A3 (ANXA3), C-type lectin domain family 4 member M (CLEC4D), Lamin B (LMNB1), proline rich and Gla domain 4 (PRRG4), TNF alpha induced protein 6 (TNFAIP6), Lamin B1 (TNFAIP6), Vanin 1 (VNN1), and Interleukin 2 receptor subunit beta (IL2RB) that participate in tumour initiation and development could act as novel biomarkers for early detection of CRC . In lineGEP has been investigated for its potential to predict the outcome of patients in other cancers, too. Méndez et al. (2011) identified five genes (Receptor accessory protein (1REEP1), Ring finger protein 145 (RNF145), CTONG2002744, Myosin VA (MYO5A), and FBXO32) that were differentially expressed between node-positive and node-negative oral squamous cell carcinomas (OSCC), and suggested that this model is applicable for identification of occult metastasis in patients . SimilarAs already discussed, the GEP platform provides a unique opportunity to examine tens of thousands of different candidate genes at a given time. However, like other laboratory methods, GEP technology is limited. The data obtained from GEP are very simple and only consist of the candidate genes. GEP is not able to detect the interactions or signaling crosstalks. Therefore, other data analyses must be performed to elucidate the underlined network, for example, behind the antitumour drug resistance. Thus, different perceptions may be derived from a single raw data point. The other limitation of the GEP approach is the sample type and the method of preparation. The DNA of different types of cells and tissues of the body undergo multiple modifications and express various genes; hence, the quality of DNA extraction and isolation is directly related to the GEP output. This noise is usually resolved by increasing the sample size, but in particular cases, other alternatives such as RNA sequencing technology ,53 or siOverall, for prediction of tumour recurrence and metastasis, GEP analysis has marked advantages compared to routine clinical exams. Using microarray technology, various genes are identified that allow prediction of LNM in CRC cases. Based on this approach, a new classification of CRC has been introduced that reflects the different biological pathways and distinct prognostic features, allowing preselection of patients who would benefit from adjuvant therapies. However, more investigations are also needed to identify the genes associated with poor prognosis profiles, since these may actually prove interesting potential targets for rational development of new cancer drugs.


Arabidopsis thaliana is the most widely used model organism for research in plant biology. While significant advances in understanding plant growth and development have been made by focusing on the molecular genetics of Arabidopsis, extracting and understanding the functional framework of metabolism is challenging, both from a technical perspective due to losses and modification during extraction of metabolites from the leaves, and from the biological perspective, due to random variation obscuring how well the function is performed. The purpose of this work is to establish the in vivo metabolic profile directly from the Arabidopsis thaliana leaves without metabolite extraction, to reduce the complexity of the results by multivariate analysis, and to unravel the mitigation of cellular complexity by predominant functional periodicity. To achieve this, we use the circadian cycle that strongly influences metabolic and physiological processes and exerts control over the photosynthetic machinery. High resolution-magic angle spinning nuclear magnetic resonance (HR-MAS NMR) was applied to obtain the metabolic profile directly from intact Arabidopsis leaves. Combining one- and two-dimensional 1H HR-MAS NMR allowed the identification of several metabolites including sugars and amino acids in intact leaves. Multivariate analysis on HR-MAS NMR spectra of leaves throughout the circadian cycle revealed modules of primary metabolites with significant and consistent variations of their molecular components at different time points of the circadian cycle. Since robust photosynthetic performance in plants relies on the functional periodicity of the circadian rhythm, our results show that HR-MAS NMR promises to be an important non-invasive method that can be used for metabolomics of the Arabidopsis thaliana mutants with altered physiology and photosynthetic efficiency. Arabidopsis thaliana play a central role in understanding biological functions across plant species and in characterizing phenotypes associated with genetic mutations  have been overlaid for assignment purpose.(TIF)Click here for additional data file.S3 FigReference spectra of glucose (purple), sucrose (green) and fructose (red)  have been overlaid for assignment purpose.(TIF)Click here for additional data file.S4 Figα-glucose and β-glucose has been overlaid on experimental data with purple colour.Reference spectra of (TIF)Click here for additional data file.S5 FigLeaves were collected at different time points throughout 24h light/dark period (Time points are same as mentioned in (TIF)Click here for additional data file.S1 TableHR-MAS NMR conditions used in earlier studies for obtaining metabolic profile in various plant materials.(DOCX)Click here for additional data file.


With the rapid development of computer science and technologies in recent years, there has been dramatic advance in anterior segment imaging.As we have mentioned in the call for papers for this special issue, manuscripts are covering the topics of anterior segment imaging techniques including anterior segment optical coherence tomography (OCT), specular microscopy, corneal topography, confocal microscopy, and ultrasound biomicroscopy (UBM). These techniques have enabled precise visualization and objective assessments of anterior segment structures; thus, these devices have become essential tools for better diagnosis and treatment of anterior segment diseases including corneal disorders, cataract, glaucoma, and even disorders of the lacrimal system. Future development of novel modalities, that is, en face OCT or ultrahigh-resolution OCT, is expected to allow more detailed visualization at a microscopic level, which would provide even more understanding of anterior segment pathology at a cellular level. In this special issue, the authors contributed 15 original articles and 4 review papers regarding technologies of the anterior segment imaging and clinical application of the imaging devices.The authors have contributed the results of their original researches on various topics on anterior segment imaging: (1) agreement between gonioscopic examination and swept source OCT imaging; (2) preliminary outcomes of accelerated corneal collagen cross-linking using topography-guided ultraviolet-A energy emission; (3) anterior segment changes before and following laser peripheral iridotomy of iris bombe in patients with uveitic secondary glaucoma; (4) anterior segment measurements with swept source OCT before and after ab interno trabeculotomy; (5) effects of a new type implantable collamer lens implantation on vision-related daily activities; (6) corneal epithelial remodeling after transepithelial photorefractive keratectomy for myopia; (7) measurement of anterior chamber volume in cataract patients using swept-source OCT; (8) biomechanical findings in contact lens-induced corneal warpage; (9) quantitative analysis of lens nuclear density using OCT with a liquid optics interface; (10) rotating Scheimpflug imaging indices in different grades of keratoconus; (11) evaluation of outflow structures in vivo after the phacocanaloplasty; (12) evaluation of corneal epithelial thickness in asymmetric keratoconic eyes and normal eyes using Fourier domain OCT; (13) central corneal thickness measured using spectral-domain OCT and associations with ocular and systemic parameters; (14) anterior segment changes in pseudophakic eyes after pars plana vitrectomy with silicone oil or gas tamponade using UBM imaging; and (15) UBM comparison of ab interno and ab externo intraocular lens scleral fixation.This special issue also includes review articles on the following topics: (1) application of Scheimpflug imaging in glaucoma management; (2) anterior segment imaging in ocular surface squamous neoplasia; (3) applications of anterior segment OCT in cornea and ocular surface diseases; and (4) the role of anterior segment OCT and UBM in corneal and conjunctival tumors.We hope these articles will provide readers with useful information on anterior segment imaging and new ideas for researches on related topics.


Multicellular organisms rely on the precise and consistent regulation of gene expression to direct their development in tissue- and cell-type specific patterns. This regulatory activity involves arrays of DNA-binding transcription factors and epigenetic factors that modify chromatin structure. Among the chromatin modifiers, trithorax (trxG) and Polycomb (PcG) group proteins play important roles in orchestrating the stable activation and repression of gene expression, respectively. These proteins have generally antagonistic functions in maintaining cell and tissue homeostasis as well as in mediating widespread transcriptional reprogramming during developmental transitions. Plants utilize multiple trxG factors to regulate gene transcription as they modulate their development in response to both endogenous and environmental cues. Here, I will discuss the roles of trxG factors and their associated proteins in post-embryonic plant development. The development of multicellular organisms is driven by precise patterns of gene transcription that are tightly regulated in a spatial and temporal manner. Establishing and sustaining specific transcription states at gene loci are complex, multi-step processes. They require repertoires of sequence-specific DNA-binding transcription factors as well as epigenetic factors that alter chromatin structure and thereby affect accessibility by the transcriptional machinery. Epigenetic regulators classified as trithorax group (trxG) and Polycomb group (PcG) factors are critical for maintaining the stable transcription patterns at developmental regulatory loci by organizing chromatin in an active or inactive state, respectively . trxG anDrosophila as positive regulators of PcG developmental target genes (Table 1). The chromatin remodeling proteins include members of the SWI/SNF, ISWI, and CHD families that utilize ATP to alter nucleosome assembly and distribution  AP2 domain transcription factor (TF) genes, which are essential for root stem cell niche maintenance  maintenance. The RAM generates the entire underground root system and has a stereotypical organization. Four rarely dividing cells known as the quiescent center (QC) act as a niche  that maintenance .Figure 1A). The SET domain protein SET DOMAIN GROUP 2 (SDG2) is the major H3K4 trimethyltransferase in Arabidopsis and is necessary for genome-wide H3K4me3 deposition  and BRAHMA (BRM) also regulate RAM activity in Arabidopsis. PKL acts antagonistically to the PRC2 PcG factor CURLY LEAF (CLF) to maintain RAM stem cell activity -CLAVATA (CLV) signal transduction pathway. The homeobox TF gene WUS is expressed in the core of the meristem and confers stem cell identity on the overlying cells . In contrast, the SAND domain-containing proteins ULTRAPETALA1 (ULT1) and ULT2 restrict shoot and floral stem cell activity by limiting the size of the WUS expression domain , FLOWERING LOCUS D (FD) and SOC1, which promote the transition to flowering  and APETALA1 (AP1), which confer floral meristem (FM) identity on the primordia that form on the flanks of the primary SAM  to reproductive organs (flowers). The timing of this dynamic meristem cell fate switch is critical for plant reproductive success and occurs in response to endogenous pathways such as the age, GA and autonomous pathways (APs) as well as environmental cues including photoperiod, vernalization and temperature . The MADlowering . FT protmary SAM .FLC, SOC1 and FT are crucial to the timing of the floral transition, and a number of trxG factors are involved in these processes . Because the role of epigenetic factors in FLC regulation during vernalization has been extensively reviewed  couple removal of repressive histone marks with deposition of active marks and transcription initiation/elongation to sustain high levels of FLC expression. ATXR7/SDG25 also represses the floral transition by binding to the FLC promoter and augmenting both H3K36me3 and H3K4me3 accumulation , a core component of the Arabidopsis COMPASS-like complex (AtCOMPASS) that also binds the FLC locus and promotes its expression by elevating H3K4me3 levels   . SVP forSE (SVP) . SVP expcription . BRM repactivity . Thus thFLC transcription is crucial for the transition from the vegetative to the reproductive state. The JmjC domain-containing H3K27 demethylase REF6/JMJ12 promotes flowering independently of photoperiod by repressing FLC transcription  that produces floral meristems (FMs) instead of leaves. A small suite of floral homeotic transcription factor genes then specifies the identity of each floral organ – sepals, petals, stamens, and carpels – from the outside to the inside of the flower . The actAPETALA3 (AP3) and AGAMOUS (AG) that specify petal, stamen, and carpel identity in the developing flower , which recruit SYD to the AP3 and AG loci  by activating AP3 and AG expression, antagonizing CLF activity at the two loci by reducing H3K27me3 deposition and enhancing H3K4me3 deposition (The plant specific TF LFY and the MADS domain TF SEPALLATA3 (SEP3) play crucial roles in activating the expression of MADS box-containing floral homeotic genes such as g flower . SYD and AG loci . At the  AG loci   enhance ult1 FM phenotypes, and ALP1 promotes floral organ identity gene expression in the absence of LFY . It is becoming clear from recent research that members of all of these categories of proteins play important roles in regulating landmark post-embryonic developmental processes such as meristem maintenance and floral induction. Moreover some trxG factors, such as BRM and ATX1, mediate multiple developmental processes during plant growth whereas others appear to have more restricted roles. The coupling of a core set of reiteratively used trxG components together with stage-, tissue- or process-specific trxG components may provide a flexible mechanism for tailoring the basic process of transcription activation to discrete gene networks in response to changing endogenous and environmental signals during the life cycle.A variety of trxG factors exist in plants that carry out diverse biochemical activities to promote active gene expression states: chromatin-remodeling ATPases, histone methyltransferases, AtCOMPASS core components, histone demethylases, as well as DNA-binding and accessory proteins (Although significant progress has been made in determining the roles of trxG factors in plant development, many gaps in our understanding remain. It still remains to be determined how many trxG complexes exist in plants, not to mention their full composition and whether that composition is static or changes depending on the developmental stage or tissue. The DNA binding proteins that recruit trxG factors to developmental regulatory loci are only beginning to be identified, while elucidating the chromatin signatures of plant stem cell populations can provide a valuable starting point for determining how tissue- and stage-specific epigenetic states are ultimately achieved during development. Finally, much work remains to decipher how developmental switches between trxG and PcG activities are implemented at individual loci as well as broadly across the genome to coordinate widespread transcriptional reprogramming. Further investigation in these areas will provide a more complete picture of how plants are able to maintain and as necessary adjust their gene expression programs during development in response to a wealth of endogenous and environmental cues.The author confirms being the sole contributor of this work and approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Introduction: Considering importance of fatty acids in developing coronary artery disease (CAD) and lack of information about saphenous vein which is commonly used as coronary arterial bypass, in this study we investigated differences in fatty acids composition between saphenous vein and aorta tissues in patients with CAD.Methods: Biopsy samples of aortic tissues and saphenous veins as well as blood samples were obtained form 42 patients with CAD. Fatty acids composition of the tissues was determined using gas chromatography and also serum lipid profile was evaluated by commercial kits.Results: Levels of palmitic acid (16:0) were significantly higher in aorta in compared with saphenous (P < 0.001). Also levels of most unsaturated fatty acids  were statistically higher in saphenous tissue than aorta tissue (P < 0.05). Mean levels of linoleic acid (18:2 n-6) was higher in aorta tissue in comparison with saphenous tissue (P = 0.01). We observed positive correlations between serum levels of LDL-C with elaidic acid and linoleic acid levels in saphenous. Evaluation of aorta tissue fatty acids revealed that palmitoleic acid (16:1) had positive and arachidonic and linoleic acids had negative correlations with serum HDL-C levels.Conclusion: Our results revealed difference between fatty acids composition of aorta and saphenous vein tissues and existence of correlations between the fatty acids levels with serum lipid profile. The saphenous vein had higher poly-unsaturated fatty acids in compared to aorta tissue and thus this vein is not at risk of atherosclerosis and can be used as coronary arterial bypass. Although many studies have evaluated effects of dietary fatty acids as well as plasma levels of them on coronary atherosclerosis,12 but there is no enough information about fatty acids content and their possible roles in the tissues which are involved in the atherosclerosis.Many studies have been conducted to investigate lipid accumulation in the arterial wall of patients with atherosclerosis but many questions about mechanisms of dyslipidemia and progression of atherosclerotic lesions have been remained unanswered.14 However existence of higher free cholesterols than cholesterol esters in the plaques15 which is in contrary with the finding about high ratio of cholesterol esters/free cholesterol in plasma,16 led to arise theory of coming the lipids from vessels growing into the plaque.17 On the other hand higher levels of fatty acid synthesis in atherosclerotic arteries in compared with intact arteries highlights possible role of fatty acid metabolism in tissues which are involved in atherosclerosis.18It has been reported that cholesterol as one of the major lipids in atherosclerotic plaques which possesses fatty acids, enters from the blood rather than in situ synthesis.19 Also Bahrami et al20 have demonstrated lower levels of unsaturated fatty acids and higher amounts of saturated fatty acids in atherosclerotic aorta in compared with the internal mammary arteries. Furthermore different fatty acids composition of aorta artery and adipose tissue between patients with and without CAD has been shown previously.21 These finding emphasize on importance of local fatty acids composition in developing atherosclerosis and CAD. However there is no information about fatty acids composition of saphenous vein which is commonly used as coronary arterial bypass and also has structural differences with arteries.22 The aim of this study was to investigate differences in fatty acids composition between saphenous vein and aorta tissues, as well as associations between the fatty acids levels with serum lipid profile in patients undergoing CABG.In a study it has been reported that the fatty acids composition of coronary arteries was significantly different between sudden cardiac death and traffic accident victims.2 and the patients with class II and III obesity (BMI ≥39 kg/m2) were not involved. The exclusion criteria were: having acute inflammation, auto-immune diseases, kidney and liver diseases, diabetes, anemia or malignant tumors. Also patients with prior CABG as well as aortic or mitral valve replacement were excluded from the study.Frothy-two individuals (mean age: 53.2±7.5 years), documented as CAD patients were included in this study. All patients had at least two vessels coronary artery disease (≥50% stenosis) based on the angiographic data and were scheduled for CABG at Shahid-Madani heart hospital in Tabriz, a northwest city of Iran. Also we recruited patients with body mass index (BMI) ≤34.9 kg/mBiopsy samples (1-2 g) were obtained during the surgery from aortic tissues and saphenous veins and then were dissolved in hexane and kept frozen at - 70°C in glass vials for fatty acid measurement. Also in order to lipid profile analysis, approximately 5 mL of venous blood samples in the fasting state at 8:00 am were taken from all participants and the serums were separated using centrifugation.23 and the percentage of each fatty acid was determined using gas chromatography. In brief, the solution of fat extract in hexane was evaporated under a stream of nitrogen to near dryness, and the lipids were esterified with methanol during catalysis with acetyl chloride. Then fatty acid methyl ester derivatives were separated on a 60- x 0.25-mm Teknokroma TR-CN100 column using gas chromatograph system  equipped with a split injector and a flame ionization detector. The oven temperature program was 170-210°C, 1°C/min, and then isothermal for 45 minutes. The combined inter- and intra assay of variation was 2.1% for palmitic acid (16:0), 2.7 for linoleic acid (18:2n-6), 18.6 for eicosapentaenoic acid (EPA), and 11.2 for docosahexaennoic acid (DHA). The known fatty acids from Sigma chemicals was employed as standards to peak retention times identification. The levels of fatty acids were calculated as the percentage of each fatty acid of the total.Fatty acids were extracted using the Bligh and Dyer methodThe levels of triglycerides (TG), total cholesterol (TC) and high density lipoprotein-cholesterol (HDL-C) determined by enzymatic colorimetric method  with an automated chemical analyzer . Low-density lipoprotein-cholesterol was calculated using the Friedewald formula.t test was used to compare fatty acids levels between saphenous vein and aorta tissues in each patient. Also the Pearson correlation test was applied to investigate possible associations between the fatty acids levels with serum lipid parameters. P values <0.05 were considered significant. SPSS V.16 software was used for the statistical analysis.The data are reported as mean ± standard deviation (SD). Normal distribution of the data was confirmed by Kolmogorov-Smirnov test. The paired Demographic and clinical characteristics of the studied population are shown in In studied patients mean levels of total cholesterol was higher than normal range (214 ± 45 mg/dL) and mean levels of triacylglycerol, 178 ± 50 mg/dL, was borderline high . Also thP < 0.001). Levels of most unsaturated fatty acids  were statistically higher in saphenous tissue than aorta tissue (P < 0.05). On the other hand, mean levels of linoleic acid (18:2 n‐6) was higher in aorta tissue in comparison with saphenous tissue . Comparing the mono-unsaturated fatty acids levels between saphenous and aorta demonstrated that levels of palmitoleic acid and oleic acid as well as its trans isomer (elaidic acid) were significantly lower in aorta than saphenous vein. The poly-unsaturated fatty acids levels were also different between the two vessels as we observed significantly lower proportions of the trans isomer of linoleic acid (18:2 t), linolenic acid (18:3 n-9) and docosatrienoate acid (22:3 n-3) in aorta in compared to the saphenous. Based on our knowledge, there is no report on differences of fatty acids composition between saphenous and aortic tissues of CAD patients. However, in accordance with our results, in previous studies higher levels of palmitic acid and also lower levels of poly-unsaturated fatty acids in phospholipids of aorta and coronary arteries of patients with CAD in compared with control individuals have been reported.21 Moreover in a study on fatty acids composition of aorta and internal mammary of patients with CAD same as our results, lower levels of unsaturated fatty acids  and higher percentage of palmitic acid in aorta samples than internal mammary have been reported.20 Moreover they also found statistically different levels of stearic acid (18:0), arachidonic acid (20:4n-6) and eicosapentaenoic acid (20:5-n3) between aorta and internal mammary arteries.20 We did not find such differences between aorta and saphenous vein maybe due to structural differences between saphenous vein and internal mammary artery.The assessment of fatty acids contents in saphenous vein and aorta of CAD patients indicated that the fatty acids composition between these vessels was different. We found higher concentration of palmitic acid as saturated fatty acid in aorta in compared with saphenous vein  were found in aorta tissue as compared to the saphenous vein. Higher levels of linoleic acid in aorta and coronary arteries of individuals with CAD compare to non-CAD subjects and also in aorta in compared with internal mammary artery in patients with CAD have been shown previously.20 However there is no enough information about pattern of fatty acids modification and its role in atherosclerosis pathogenesis.Such differences between fatty acids composition of aorta artery and saphenous vein could be partly due to structural differences between arteries and veins but some part of these differences could also be because of atherosclerosis and cardiovascular complications that the aorta was involved. Reports about different fatty acids composition between aorta and internal mammary of patient with CAD confirm this possibility.33 and thromboxane A2 production.34 In our previous study we also found negative correlations between levels of linoleic acid in epicardial and subcutaneous adipose tissues with serum levels of HDL-C and a positive correlation between this fatty acid content in subcutaneous adipose tissue with serum LDL-C in patients with CAD.35 There are wide evidences that the HDL-C and LDL-C are protective and risk factors for atherosclerosis and its complications, respectively37 and on the other hand linoleic and arachidonic acids have been documented as atherogenic fatty acids which cause inflammation and platelet aggregation.32 So finding the associations between these fatty acids with LDL-C and HDL-C seems reasonable. Moreover, we observed a positive correlation between the HDL value and palmitoleic acid in aorta which confirms the protective effect of palmitoleic acid as MUFAs against CAD.39In this study we detected inverse correlations between serum HDL-C with linoleic acid levels in saphenous and aorta tissues and also with arachidonic acid levels of aortic tissue. Besides, linoleic acid levels in saphenous vein were positively correlated with LDL-C in serum. The negative correlation between HDL-C and arachidonic acid could be explained by results of previous study reported that in CAD patients, HDL-hypocholesterolemia decreases ability of platelet to produce nitric oxide (NO) and consequently reduction in NO levels itself inhibits arachidonic acid metabolism. NO induces the inhibitory influence through various direct and indirect ways such as inhibition of cyclooxygenase activities40 Moreover results of a 25-year follow up study conducted in seven countries, have been showed positive correlation between dietary intake of elaidic acid and serum cholesterol.41 These reports confirm our finding about existence of positive correlation between elaidic acid in saphenous and serum total cholesterol.It has been demonstrated that elaidic acid (18:1t) is cholesterol elevating factor.43 exerts its effects partly via changing serum lipid profile and consequently vessel wall fatty acids composition. Although observed differences in fatty acids composition between aorta and saphenous vein tissues and also between aorta tissues of CAD and NON-CAD patients21 as well as aorta and internal mammary arteries of CAD patients20 emphasize that in addition to diet, local lipid metabolism and existence of atherosclerosis can be determinative factors for fatty acid composition of vessel walls. However the ways that different dietary intake or lipid metabolism could influence tissue fatty acids composition still remain to be elucidated. However, in our study we did not have information about dietary habits and life-style which could possibly affect the fatty acids composition of the tissues. Although it should be noted that in the present study the comparisons were done between two tissues of each patient, so any confounding factor such as dietary habits which could affect the evaluated parameters, affects the parameters of both saphenous and aorta tissues in same patient and consequently could not affect the overall result of our study. Another limitation of our study was lack of proper control without CVD because of impossibility of taking such samples from healthy individuals. However further studies could be conducted on immediately dead persons without CVD or animal models.Considering the association between fatty acids composition of aorta and saphenous tissues with serum lipid profile that was found in the present study, it can be concluded that composition of dietary fatty acids which has been suggested as important factor for CAD developmentIn this study due to the unavailability of healthy individuals saphenous and aorta tissues we could not compare the results with control. Also due to difficulty in finding patients our study was conducted with almost small sample size. Another limitation of our study was lack of information about diet of patients as it can has effect on fatty acids composition of saphenous and aorta tissues.In conclusion our results revealed difference between fatty acids composition of aorta and saphenous vein tissues in CAD patients and existence of correlations between the fatty acids levels and serum lipid profile. Owing our results about higher levels of PUFAs in saphenous vein compared to aorta tissue, it is likely that the vein has almost low risk for atherosclerosis and may be able to be used as coronary arterial bypass.All authors declare no competing financial interests exist.Approval for this study was granted by the ethical committee of Tabriz University of Medical Sciences and the informed written consent was obtained from all participants The work was supported by a grant from the Cardiovascular Research Center, Tabriz University of Medical Sciences. This article was extracted from the M.Sc. thesis of Mirhamid Paytakhti Oskouei.


A patient aged 58 years, presented with 2 months history of an erythematous, painful and bleeding nodule, after labial herpes (A). The dermoscopic examination confirms the diagnosis of a pyogenic granuloma (B). The dermoscopic criteria are: the reddish homogeneous area (green line) whose color varied from completely red to red with whitish zones which is corresponded to proliferating vessels. A white collarette (yellow line) corresponds to the hyperplastic adnexal epithelium that embraces the lesion at the periphery. The white lines (blue line) like a double rail which is corresponded to a fibrous septa that criss-cross the lobules. And finally ulceration or hemorrhagic crusts (black line) which are frequently observed in pyogenic granuloma.


Many alien plants use animal vectors for dispersal of their diaspores (zoochory). If alien plants interact with native disperser animals, this can interfere with animal-mediated dispersal of native diaspores. Interference by alien species is known for frugivorous animals dispersing fruits of terrestrial plants by ingestion, transport and egestion (endozoochory). However, less attention has been paid to possible interference of alien plants with dispersal of diaspores via external attachment , interference in aquatic ecosystems, or positive effects of alien plants on dispersal of native plants. This literature study addresses the following hypotheses: (1) alien plants may interfere with both internal and external animal-mediated dispersal of native diaspores; (2) interference also occurs in aquatic ecosystems; (3) interference of alien plants can have both negative and positive effects on native plants. The studied literature revealed that alien species can comprise large proportions of both internally and externally transported diaspores. Because animals have limited space for ingested and adhering diaspores, alien species affect both internal and external transport of native diaspores. Alien plant species also form large proportions of all dispersed diaspores in aquatic systems and interfere with dispersal of native aquatic plants. Alien interference can be either negative  or positive . I propose many future research directions, because understanding whether alien plant species disrupt or facilitate animal-mediated dispersal of native plants is crucial for targeted conservation of invaded (aquatic) plant communities. Dispersal of plant diaspores by animals (zoochory) is a globally important mechanism regulating species diversity of plant communities. Key disperser species include birds, mammals, and fish–of which many forage on plant diaspores, transport them internally during digestion, and defecate viable diaspores in new locations  alien plant species may interfere with both internal and external animal-mediated dispersal of native diaspores; (2) interference by alien plants also commonly occurs in aquatic ecosystems; and (3) interference by alien plants can have both negative and positive effects on native plants. I focus on zoochory of plant diaspores  by vertebrate animals. “Alien species” is used as the terminology throughout this study to refer to all non-native, invasive, introduced or exotic species that established a self-sustaining reproductive population outside their native geographic area  feed selectively on Eurasian grasses in Canada  and mule deers (Odocoileus hemionus) remove fruits of an alien succulent faster than those of a native species in the same genus  studying interference of alien plants with ectozoochory by itself, and simultaneously with interference of endozoochory in single model systems; and (2) comparing the relative effectiveness of alien and native diaspores for endo- and ectozoochory in choice and attachment experiments  AND TS = (seed OR diaspore* OR fruit OR nut); combined with either TS = (frugivor* OR endozoochor*) or with TS = (ectozoochor* OR epizoochor* OR exozoochor*). This resulted in respectively 376 and 36 studies that addressed dispersal of alien plants for endo- and ectozoochory. Addition of the term TS = (aquatic OR wetland OR freshwater OR riparian) reduced the publication counts to respectively 16 and 6, indicating that a low percentage of all studies involved aquatic habitats.To compare study effort on alien interference between habitats and among disperser animals, I performed a standardized search for publications reporting the presence of alien and native aquatic diaspores in single samples of feces or attached to native vertebrate animals in ISI Web of Science. On the 11th of January 2018 I entered the following search string: TS =  and 55% for ectozoochory . In 5 of the 19 reports on endozoochory and 8 of the 15 reports of ectozoochory more alien than native diaspores were dispersed. However, these numbers should be interpreted with caution because variation among geographic locations and studied species was large, and studies reporting only dispersal of native diaspores were not included.All aquatic studies jointly reported 34 cases of potential interference by alien plants with dispersal of native diaspores—by native disperser animals—and in aquatic ecosystems Tables , 2. The The presented cases illustrate that patterns of alien interference thus far primarily described in terrestrial ecosystems may also apply to aquatic ecosystems. Future research should focus on under-examined disperser animals like fish, bats and reptiles  although the phenomenon has been primarily studied for endozoochory by frugivorous birds and mammals in terrestrial ecosystems, alien species may also interfere with ectozoochory and this warrants further studying; (2) interference of alien species with zoochory can similarly be found in aquatic ecosystems; (3) alien plant species can also provide resources such as food or nesting habitat to animals, which can increase densities of native disperser animals or attract new disperser animal species. Through these mechanisms alien plants can also positively affect dispersal of native plants, which warrants further studying. This study illustrates that the impacts of alien plant species on native plant species, whether positive or negative, can vary among native plant species relying on different dispersal mechanisms. Understanding species interactions is crucial for effective conservation, especially in invaded ecosystems.CvL designed the study, collected and analyzed the data, and wrote the manuscript.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


To respond to the World Health Assembly call for dissemination of lessons learnt from countries that have begun implementing the International Health Regulations, 2005 revision; IHR (2005).In November 2015, we conducted a systematic search of the following online databases and sources: PubMed®, Embase®, Global Health, Scopus, World Health Organization (WHO) Global Index Medicus, WHO Bulletin on IHR Implementation and the International Society for Disease Surveillance. We included identified studies and reports summarizing national experience in implementing any of the IHR (2005) core capacities or their components. We excluded studies that were theoretical or referred to IHR (1969). Qualitative systematic review methodology, including meta-ethnography, was used for qualitative synthesis.We analysed 51 articles from 77 countries representing all WHO Regions. The meta-syntheses identified a total of 44 lessons learnt across the eight core capacities of IHR (2005). Major themes included the need to mobilize and sustain political commitment; to adapt global requirements based on local sociocultural, epidemiological, health system and economic contexts; and to conduct baseline and follow-up assessments to monitor the status of IHR (2005) implementation.Although experiences of IHR (2005) implementation covered a wide global range, more documentation from Africa and Eastern Europe is needed. We did not find specific areas of weakness in monitoring IHR (2005); sustained monitoring of all core capacities is required to ensure effective systems. These lessons learnt could be adapted by countries in the process of meeting IHR (2005) requirements. First introduced in 1969, IHR is global legislation requiring countries to link and coordinate specific actions.,,IHR (2005) also requires all countries to develop, strengthen and maintain eight core public health capacities.Although IHR (2005) describes what must be achieved by countries, there is limited knowledge on how countries should proceed in achieving the core capacities. To fill this gap and accelerate implementation of IHR (2005), the World Health Assembly in 2015 identified a need to evaluate and share the lessons learnt from countries that have implemented IHR (2005).,,This systematic review was conducted in accordance with Enhancing Transparency in Reporting the Synthesis of Qualitative Research guidelines, using a predefined protocol .Box 1Component 1A: National legislation and policyComponent 1B: FinancingComponent 2A: IHR coordination, communication and advocacyComponent 3A: Indicator-based surveillanceComponent 3B: Event-based surveillanceComponent 4A: Rapid response capacityComponent 4B: Case managementComponent 4C: Infection controlComponent 4D: Disinfection, decontamination and vector controlComponent 5A: Public health emergency preparedness and responseComponent 5B: Risk and resource management for IHR preparednessComponent 6A: Policy and procedures for public communicationsComponent 7A: Human resource capacityComponent 8A: Policy and coordination of laboratory servicesComponent 8B: Laboratory diagnostic and confirmation capacityComponent 8C: Laboratory biosafety and laboratory biosecurity (biorisk management)Component 8D: Laboratory-based surveillanceIHR: International Health Regulations.Source: International Health Regulations (2005).Two authors independently screened all abstracts, articles and reports and then matched the full texts selected during screening against the inclusion criteria. The reference lists of relevant articles and reviews were also searched for additional studies and reports. Articles meeting the inclusion criteria were included in the review. Three authors completed the data extraction using standardized extraction tables.,Two authors independently read and assessed the quality of included studies using the Critical Appraisal Skills Programme qualitative research checklist.,–,We used the Cochrane qualitative systematic review method, including meta-ethnography, to synthesize qualitative data.–http://www.who.int/bulletin/volumes/96/2/16-189100). A more detailed summary of the included articles and themes identified is available from the corresponding author. Some articles reported national experience in IHR (2005) implementation in multiple countries. In total, 77 countries were represented from all WHO Regions: 23 from Western Pacific, 16 from Europe, 14 from Africa, 11 from the Americas, eight from Eastern Mediterranean and five from South-East Asia  from central government, subnational levels of government, civil society and the scientific community.,,2. Developing appropriate policies and allocating responsibilities for the workforce is needed at both central and subnational levels of government.,,3. Legislation and policies should strive to incorporate and balance global, regional, national and subnational priorities for health security.,,4. For sustainable financing, detailed long-term national strategic plans with budget allocations should be developed in partnership between government, national stakeholders, and non-state actors .,5. The national health system and sociocultural context of a country will determine priorities for funding and other resource needs for IHR (2005) core capacity development, emergency response and the health workforce.,,6. Creating and empowering a national secretariat responsible for mobilizing and securing resources from state and non-state actors can help ensure sustainable and continuous financing.,,1. Existing regional and global intergovernmental organizations can be recruited to collaborate and co-manage international epidemics.,,2. A strong programmatic structure at the national and subnational levels, including intersectoral national technical working groups, can assist with comprehensive and effective coordination.,,3. Establishing roles, responsibilities and terms of reference of national focal points within national legislation, and disseminating information inside government, to national stakeholders, and to WHO, improves coordination and communication.,,,4. Adequately resourced communication channels and standard operating procedures, from the subnational to the inter-governmental level, facilitate the timely transmission and retrieval of information.,1. It is important to create and empower a national government entity to coordinate disease surveillance and control efforts.,2. Strategic plans for disease surveillance at both the national and subnational levels can enable long-term sustainability of these efforts.,,3. Key communicable diseases need to be categorized based on epidemiological context .,4. Surveillance at the subnational level can be sustained by developing legislation that outlines the requirements, mechanisms  can develop capacity for collection, analysis and use of data at subnational levels.,,6. A unified and dynamic surveillance system to track alerts, updates and early warnings and response, and using technologies appropriate to the national context , encourages appropriate information use and accurate reporting.,,,7. Standardizing passive data collection, management and validation procedures across all reporting sites, and actively searching for potential surveillance intelligence through local communication channels , improves the usability and timeliness of data.,1. Defining criteria for an emergency public health event  assists with the initiation of an appropriate and timely response.,,,2. Establishing roles, responsibilities and terms of reference for a multidisciplinary response unit supports coordination of public health emergency responses at the national and subnational levels.,,3. Including national and international stakeholders in the development of guidelines for rapid and emergency response facilitates wide technical ownership and financial support.,4. Designing and resourcing health facilities to minimize nosocomial infection transmission improves the efficacy of response without encouraging further spread .Box 3,1. Comprehensive national and subnational budgeted plans for preparedness of emergency public health events are needed to support timely action.,2. Exercises, drills, assessments and evaluations can test capacity to implement the response plans and ensure they are adequate and operational.,,3. A national reserve of medicines, vaccines and laboratory supplies and reagents helps with management of public health emergencies.,,1. An intersectoral system for risk communication of public health emergencies, functioning at all levels of government, ensures timely action and response.,,2. Risk communication systems should be bidirectional, with linkages to national and international public health partners, to collect and disseminate information about threats.,,3. Regular and emergency information to national and international stakeholders assists in building and maintaining effective communication channels and relationships.,,4. Standard operating procedures ensure personnel are ready to investigate and communicate emerging epidemics and support preparedness for activation of early warning systems.,,,5. Using a combination of media to send and receive information ensures wide access and use.,,1. Identifying competency gaps in the workforce ensures that training and resources are targeting the highest priority needs .,,2. Selecting appropriate media for developing competencies based on health system context and social and cultural preferences can increase the efficacy of training.,,3. Establishing workforce oversight mechanisms for long-term maintenance of current and future needs assists with sustainability of individual training efforts.,,4. Nationally agreed strategies to recruit, train and retain health workforce can help support IHR (2005) competency development at all levels of the health system.,,,,1. It is important to create a governmental entity at the national level to coordinate national strategic plans for laboratory response and develop national standards, guidelines and operating procedures for laboratory surveillance at national and subnational levels.,,2. Regional and global reference laboratories can be helpful when developing laboratory surveillance networks.,,3. Validated criteria and mechanisms to accredit laboratories for core surveillance tasks help ensure high-quality testing.,4. Laboratory training, routine assessments and feedback, and quality assurance systems can help improve surveillance capacity and function of national and subnational laboratories.,5. Bidirectional flows of data, specimens and communication between national and subnational levels of government improve laboratory system effectiveness and adequate public health investigation.,,6. Specific policies, strategies, regulations and standard operating procedures for high-containment biosafety laboratories help ensure the safety and efficacy of laboratory testing.,,1. Conducting a baseline needs assessment can help to identify the status of a country’s health system and actions needed to meet IHR (2005) requirements.,,2. Ensuring flexible systems can improve response to known epidemics and detection of new threats.,,3. A country’s’ sociocultural, epidemiological, health system and economic context and priorities will guide national implementation of IHR (2005) core capacities.,4. Although early warning systems are part of core capacity in surveillance, they also affect coordination, risk communication and response for new epidemics.,,,5. Having a skilled workforce in place, from frontline staff to senior management, is important for ensuring sustainable expertise, implementation and commitment.IHR (2005): International Health Regulations, 2005 revision: WHO: World Health Organization. Notes: Framework for the analysis was based on the eight core capacities of the International Health Regulations (2005).,–,–,,,–,,,–,,,,–A total of 33 articles from 60 countries reported experience implementing the national legislation, policy and financing core capacity.–,,,–,–,–,,,,,,,,,,,,,Experience implementing the coordination and communications capacity was reported in 33 articles from 51 countries.–,–,,,–,,,,,,,,,,–,,,Forty articles from 63 countries reported experience implementing the surveillance core capacity.–,,,,,,–,,,,,,,Experience implementing the response core capacity was reported by 27 articles from 47 countries.,,,,–,,,,,,,,,Thirteen articles from 37 countries reported experience implementing the preparedness core capacity.–,,,,,,,–,,,,,,,,A total of 23 articles from 36 countries reported on implementing the risk communication core capacity.,,–,–,,,–,–,–,–,,–,,,,,,,,A total of 37 articles from 49 countries reported experience implementing the human resources core capacity.–,,,–,,,,–,,,,,,,,,Experience implementing the laboratory core capacity was reported in 26 articles from 44 countries.We identified five global lessons learnt that related to multiple IHR (2005) core capacities . Some maWe found substantial documentation of lessons learnt from implementing IHR (2005) core capacities globally. Although many factors affected implementation in a country, the structure of the health system, sociocultural factors, economic status and type of governance, we were able to extract some common themes. We found few gaps in implementation in direct relation to the IHR (2005) core capacities. Nonetheless, additional research could be useful in evaluating countries’ core capacity performance against actual disease epidemics. Specifically, we did not identify any documentation evaluating IHR (2005) core capacity development with national performance in preventing, detecting and responding to disease epidemics. This type of evaluation could provide useful data for future revisions of the IHR. Moreover, defining fundamental data needs for new treatments and vaccines could help develop national research capacity to shorten the development window during future disease epidemics. Finally, additional documentation of experience from Eastern Europe and Africa could help fill knowledge gaps.,Countries had different national health plans outlining how to organize their systems and strategies for managing threats and hazards.Most countries reported some experience in implementing communicable disease surveillance as part of IHR (2005).,,Although complementary in nature, the preparedness and response core capacities were often implemented independently of one another. For example, areas requiring coordination across both of these core capacities, such as case management or infection control, were not well-documented. Taking a holistic approach to IHR (2005) core capacity development could avoid these issues.,,,Risk communication focuses primarily on effective communication strategies during disease epidemics. Some of the articles we identified primarily discussed communication systems.Global strategy on human resources for health: workforce 2030.,,The World Health Assembly recently approved the –Laboratory networks are an essential part of national and regional health systems. Global laboratory networks are in place for polio, immunization programmes and influenza.–,–Given the many priorities in development, and limited national budgets to achieve them, external funding opportunities often help mobilize political commitment and national action. Experience from South Africa suggested that while external funding sources can be inflexible and unsustainable within national governance frameworks, such funding could create momentum in generating domestic financing and accelerate progress in implementing IHR (2005).,Along with financial and technical support, additional international measures could help accelerate IHR (2005) core capacity development. For example, since past disease epidemics affected regional and global economic growth, economic issues are relevant.This study has several limitations. We focused only on IHR (2005) core capacities and not on the core capabilities of points of entry, zoonotic events, food safety, chemical events and radiation emergencies.Despite considerable progress, countries that are yet to implement IHR (2005) core capacities may have insufficient human and financial resources to meet their obligations in the near future. Recent global epidemics have galvanized high-level political commitment towards ensuring global health security. We can leverage this commitment by mobilizing resources and securing wider collaboration to apply the lessons outlined here and accelerate the development of IHR (2005) core capacities globally.


Sholl analysis confirmed regular pattern of dendritic branching. The total length of dendrites was around 2100 μm with the average length of individual branching (intermediate) segment around 22 μm and for the terminal segment around 100 μm. Even though each experimental group underwent the same strictly defined protocol in tissue preparation and Golgi staining, we found inconsistency in dendritic volume and soma surface. These changes could be methodologically influenced during the Golgi procedure, although without affecting the dendritic length and tree complexity. Since the neuronal activity affects the dendritic thickness, it could not be excluded that observed volume inconsistency was related with functional states of neurons prior to animal sacrifice. Comprehensive analyses of tree complexity and dendritic length provided here could serve as an additional tool for understanding morphological variability in the most numerous neuronal population of the striatum. As reference values they could provide basic ground for comparisons with the results obtained in studies that use various models of genetically modified mice in explaining different pathological conditions that involve MSNs.In this study we have provided a detailed quantitative morphological analysis of medium spiny neurons (MSNs) in the mice dorsal striatum and determined the consistency of values among three groups of animals obtained in different set of experiments. Dendritic trees of 162 Golgi Cox (FD Rapid GolgiStain Kit) impregnated MSNs from 15 adult C57BL/6 mice were 3-dimensionally reconstructed using Neurolucida software, and parameters of dendritic morphology have been compared among experimental groups. The parameters of length and branching pattern did not show statistically significant difference and were highly consistent among groups. The average neuronal soma surface was between 160 μm The medium spiny neurons (MSNs) are the most numerous striatal neurons. Data about their morphology are needed to understand basal ganglia circuitry structure and function in normal conditions and in related disorders  kindly provided by the Max Planck Institute for Evolutionary Anthropology in Leipzig, Germany , group 2 (n = 4) and group 3 (n = 5). All work with the animals used in this study was performed in accordance with the governmental and institutional ethical guidelines and with the approval of Ethics Committee from Max Planck Institute for Evolutionary Anthropology in Leipzig and School of Medicine University of Zagreb.Brain tissue was processed separately in three sets of experiments performed at different times and defined as group 1  before euthanizing. Brains were not perfused and were removed quickly from the skull to avoid any damage to the tissue. After rinsing, the tissue was sliced in approximately 10 mm thick blocks. The blocks were stained with the FD Rapid GolgiStain™ kit . They were first immersed in the impregnation solution (A and B) which was replaced after 6–12 h and then kept in dark for 15–16 days. Afterwards, the blocks were put in Solution C which was replaced after 24 h and kept in dark for the next 48–60 h. Cryomicrotome  was used to cut 200 μm thick slices. Slices were mounted on a gelatin-coated microscope slides, stained, and dehydrated and coverslipped with Permount. The tissue obtained from group 3 was additionally treated with Toluidine blue stain before dehydration.Tissue preparation and staining were all done by the same person (U.B.) following the FD Rapid GolgiStain™ kit manufacturer’s protocol.z-axis motorized Olympus BX61 microscope equipped with x-y motorized stage guided by MAC5000 stage controller .In total 162 MSNs were three-dimensionally reconstructed using motorized microscope-computer based system and the Neurolucida software version 10 . System was composed of Only neurons with cell bodies located in the dorsal part of striatum within the middle third of the section thickness were included in the analysis. Figure By using a Neurolucida software extension NeurolucidaExplorer, from each reconstructed neuron the following parameters of somato-dendritic morphology have been extrapolated to numeric values: (1) cell surface; (2) number of primary dendrites; (3) total number of segments; (4) total dendritic length; (5) average length per dendrite; (6) average number of segments per dendrite; (7) average length and volume of individual intermediate (branch); (8) terminal; and (9) incomplete segment.For the analysis of segment length and volume, each segment was defined as branch (intermediate), terminal, or incomplete  per group using average values per neuron as raw data. We applied the STATISTICA software for statistical analysis. The dendritic variables were tested separately using one-way analysis of variance .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.


Vibrio parahaemolyticus (Vp) is found naturally in coastal saltwater. In the United States, Vp causes an estimated 35,000 domestically acquired foodborne infections annually (Vibrio Surveillance system. Vp isolates are distinguished by serotyping (>90 serotypes have been described) and by pulsed-field gel electrophoresis (PFGE).annually , of whicVp serotypes O4:K12 and O4:K(unknown) comprise the Pacific Northwest (PNW) strain and, within the United States, had not been associated with shellfish outside the Pacific Northwest before 2012. During May–July 2012, Vp of the PNW strain associated with shellfish from Oyster Bay Harbor in New York caused an outbreak of 28 illnesses in nine states. Simultaneously, Vp of the PNW strain caused an outbreak of illnesses on a cruise ship docked on the Atlantic Coast of Spain; illness was associated with cooked seafood cooled with ice made from untreated local seawater. All Vp isolates from ill persons in the U.S. and Spanish outbreaks that were further subtyped were indistinguishable by PFGE  facilitates identification of Vibrio species. Consumers can reduce their risk for Vp infection by avoiding eating raw or undercooked shellfish, especially oysters and clams.†This PNW strain is possibly becoming endemic in an expanding area of the Atlantic Ocean. The mechanisms for this introduction are not known. During the 2014


Myriad roles of mucins in normal tissues have been well documented, including lubrication of the epithelial surfaces; protection from physical damage; facilitation in cell-cell signaling and suppression of inflammatory activity. Pathological expression of mucins has been noted in cancer development and progression. This study sought to identify and quantify the types of mucins produced during various histological grades of colon cancer and to assess the diagnostic significance.Formalin fixed, paraffin-embedded tissue blocks, comprising three (3) normal colon and twenty-two (22) colon cancer tissues, were retrieved from the archives of the histopathology department of the Komfo Anokye Teaching Hospital. They were stained with Haematoxylin and eosin (H&E) for diagnosis and grading of tumours. Tissues were pre-digested with diastase and stained with Alcian blue (pH 2.5)/Periodic Acid Schiff to characterize the mucin variants present.Our findings indicated that normal colonic tissues expressed exceptionally high amount of acid mucin and low amount of neutral mucin. However, there was a general decrease in mucin expression in colon cancers compared to normal colon tissues. Additional findings suggested that as cancer progresses from low grade to high grade of adenocarcinoma of the colon, there was generally a considerable decrease in the acid mucin production and an increase in the neutral mucin expression. In contrast, a sizeable subpopulation of high-grade adenocarcinomas of colon showed a rather opposite mucin expression pattern- increase in acid mucin and a decrease in neutral mucin.As colonic cancer progresses, there are corresponding changes in the mucin types and content such that there are decrease in acid mucin and increase in neutral mucin expressions. Initiation and development of cancers, particularly colonic cancer is a very complex one with several variables playing crucial roles in sustaining the viability of the cancer. Even though initiations of colonic cancers are well attributed to genetic imbalances, which could be inherited  or somatTwenty-five formalin fixed, paraffin embedded (FFPE) colonic tissues, which were retrieved from the archives of Histopathology Unit of the Department of Pathology, Komfo Anokye Teaching Hospital, Kumasi-Ghana, following ethical approval from both the hospital and Committee for Human Research, Publication and Ethics of School of Medical Sciences, KNUST. Two tissue sections at 3μm from each FFPE tissue block were produced. One batch of the sectioned FFPE colonic tissues was stained with Haematoxylin and Eosin (H&E) and were histologically diagnosed and graded by three independent pathologists. The second batch of the sectioned FFPE colonic tissues was dually stained with Alcian blue, pH 2.5 to demonstrate acidic mucins (sulfated and carboxylated) and Periodic Acid Schiff (PAS) to demonstrate glycogens and mucins. In order to exclusively demonstrate neutral mucins, dewaxed colonic sections were pretreated with diastase for 60 minutes to digest at existing glycogen in the tissue . PositivHaematoxylin and eosin (H&E) based diagnosis and grading colonic cancer tissues: Normal colon and colonic cancer tissues were H&E stained to confirm the diagnosis and grades of the tumours. Five of the 22 colonic cancers were diagnosed and graded as Grade 2, Well Differentiated Adenocarcinoma (G2WDA); 13 of the 22 colonic cancers as Grade 2, Moderately Differentiated Adenocarcinoma (G2MDA); 3 of the 22 colonic cancers as Grade 2 Adenocarcinomas (G2A) and only 1 case as Grade 3 Poorly Differentiated Adenocarcinoma (G3PDA) ( (G3PDA) . The colDifferential mucins staining with Alcian Blue/Diastase-Periodic Acid Schiff staining: in order to correlate the mucin content/amount and types with different grading of colonic cancers, we performed Alcian blue (pH 2.5)/Diastase-Periodic Acid Schiff staining. The sensitivity and specificity of the staining mucin staining were tested using appropriate controls. This was also part of the quality assurance procedure. Quality controls for mucin demonstration For a positive control, normal colon stained blue for acid mucins using Alcian Blue (pH 2.5), Diastase Periodic Acid Schiff (DPAS) staining on prostate, and DPAS/Alcian blue (pH 2.5) staining on colon (results not shown). For a negative control, liver tissues stained negative for Alcian Blue (pH 2.5), indicating absence of acid mucins, but stained magenta in DPAS, indicating the presence of neutral mucins. Further, an omission of the periodic acid oxidation step in DPAS resulted in a negative staining on human liver (results not shown).Mucin content/amount and types in colonic cancers: in this study, the total mucin content in 22 colonic cancer cases was semi-quantified after double Alcian blue (pH 2.5) and Diastase-PAS staining. The mucin types  were also determined in each case. Both mucin content and types were estimated and correlated with tumour grade  mucins but scanty neutral mucins;It is widely postulated that sulfated mucins play a role in the control of cell division;Production of mucins, in general, is remarkably reduced in adenocarcinomas of colorectal tissues.We, in this study, demonstrated that there is a progressive decline of acid mucin from low-graded to high graded adenocarcinoma of colorectal with corresponding increase in neutral mucins;We also demonstrated for the first time that a subgroup of moderately differentiated adenocarcinomas had rather an increase in acid mucins coupled with decrease neutral mucins in humans.The authors declare no competing interest.


Based on a two-end-member mixing model, all target consumers relied on energy sources from coupled benthic and planktonic pathways, but the predominant energy source for most species was highly variable across seasons, showing seasonal trophic shift of littoral consumers. Seasonality in energy mobilization of consumers focused on two aspects: (1) the species number of consumers that relied mainly on planktonic carbon showed the lowest values in the fall and the highest during spring/summer, and (2) most consumer species showed seasonal variation in the percentages of planktonic reliance. We concluded that seasonal trophic shifts of fishes and invertebrates were driven by phytoplankton production, but benthic resources were also important seasonally in supporting littoral consumers in Meiliang Bay. Energy mobilization of carnivorous fishes was more subject to the impact of resource availability than omnivorous species.We evaluated the seasonal variation in the contributions of planktonic and benthic resources to 11 littoral predators in eutrophic Lake Taihu (China) from 2004 to 2005. Seasonal fluctuations in consumer σ


GABAergic and glutamatergic systems play an important role in the neurobiology of schizophrenia, and changes in their markers are reported in both postmortem human brain and in animal models. Recent studies have demonstrated that abnormalities in DNA methylation may underlie the alterations in various indicators of GABAergic and glutamatergic functions in schizophrenia. As our group previously found decreased NR2 protein plasma levels and downregulation of parvalbumin (PVALB) mRNA in first episode of psychosis (FEP) patients, we hypothesised that changes in DNA methylation may be responsible for these indicators of glutamatergic and GABAergic deficits in FEP patients.Blood samples were collected from patients in FEP (n = 35) after their first contact with the mental health assistance, siblings (n = 21) and population-based controls (n = 35). Bisulfite conversion and pyrosequencing were used to determine methylation levels in 4 CpG sites in promoter sequence of PVALB and 5 CpG sites at GRIN2B (gene which encodes NR2).We found hypermethylation at a CpG site within the PVALB promoter sequence in patients and their siblings compared to population-based control group (p< 0.001) while overall hypomethylation was found in the 5 CpGs analysed within GRIN2B promoter sequence (p < 0.01).Our PVALB findings are consistent with our previous studies showing that PVALB promoter methylation is elevated in schizophrenia and, additionally this is the first evidence showing changes in GRIN2B promoter methylation in psychosis. These results together suggest that these epigenetic findings may relate to the reduction of protein expression of indicators of glutamate and GABA systems seen in this disease.