Either metastatic or primary squamous cell carcinoma in the gastrointestinal tract is extremely rare, with very few cases reported in the literature. In this paper, we report a case in which the patient presented with dysphagia during the course of radiotherapy for recurrent lung cancer in a mediastinal lymph node. Although the dysphagia mimicked radiation esophagitis, the ultimate cause proved to be gastric and duodenal metastases from primary lung squamous cell carcinoma. Taking into account the value of identification of metastatic or primary SCC in the stomach and duodenum on the prognosis and treatment options, it is imperative that the correct diagnosis be established. This report is followed by a discussion of the differential diagnosis between metastatic and primary squamous cell carcinoma in the stomach and duodenum. Either metastatic or primary squamous cell carcinoma (SCC) in the gastrointestinal tract is extremely rare, with very few cases reported in the literature. The gastrointestinal tract has rarely been reported as a metastatic site of various tumors, such as lung cancer, breast cancer, hepatocellular carcinoma, melanoma, testicular seminoma, choriocarcinoma, Merkel cell carcinoma, malignant fibrous histiocytoma and others -4. Most The incidence of primary SCC of the stomach or duodenum is also very low. It is estimated that the worldwide incidence of primary SCC of the stomach is 0.04% to 0.07% . Fewer tA 54-year-old man was referred to our hospital because of a 1-month history of cough. He had smoked one pack of cigarettes per day for the past 20 years. A mass shadow in the hilum of the right lung with an enlarged subcarinal lymph node was found by contrast-enhanced computed tomography (CT) of the chest. A bronchoscopic biopsy yielded the diagnosis of SCC. After metastatic workup with abdominal ultrasonography, magnetic resonance imaging of the brain and bone scan, the patient was staged with T3N1M0 disease. Thereafter he underwent definitive right-middle lobectomy, revealing well- to moderately differentiated SCC and multiple lymph node metastases. He received four cycles of adjuvant chemotherapy comprising taxotere and cisplatin. Five months after his lobectomy, an enlarged mediastinal lymph node was discovered on a routine follow-up chest CT scan. Solitary lymph node recurrence was diagnosed by coincidence circuit single-photon emission CT assessment. The patient then underwent a course of conformal external beam radiation to the mediastinal lymph node at a dosage of 6,000 cGy in 30 fractions. He complained of dysphagia without melena or hematemesis during the fourth week of radiotherapy, which was attributed to radiation esophagitis. However, the patient’s symptoms worsened despite symptomatic treatment. A gastroduodenoscopy performed 10 days after the completion of radiotherapy revealed a giant gastric ulcer in the greater curvature of the stomach with mucosal edema and congestion as well as a huge cauliflower-like mass in the descending part of the duodenum (Figure Because of the absence of continuity between the gastric and duodenal metastatic lesions, our present case report is probably the first to describe gastric and duodenal metastases from primary lung SCC. Taking into account the value of identification of metastatic or primary SCC in the stomach and duodenum on the prognosis and treatment options, it is imperative that the correct diagnosis be established. Usually, the visual diagnosis followed by tissue diagnosis of the tumor is enough to establish the diagnosis of SCC. However, it is rarely possible to differentiate metastatic SCC from primary SCC involving the stomach and duodenum based on symptoms and imaging findings, because the major manifestations are the same. A documented history of SCC in other primary sites lends weight to the diagnosis of metastasis, especially in patients with diffuse metastases involving other organs. In this case, recurrence in the mediastinal lymph node and involvement of both stomach and duodenum were highly suggestive of metastatic rather than primary lesions. However, it could be argued that a history of malignancy does not necessarily imply that the new lesions are metastatic in nature. An early-stage primary malignancy followed by new ostensibly metastatic lesions after a relatively long interval will make the differential diagnosis of metastasis versus new primary tumor difficult. More importantly, the diagnosis of metastatic tumor to gastrointestinal tract is challenging on the rare occasions when it represents the first and only metastatic site. It would be even more challenging where the gastrointestinal involvement presents prior to the detection of the primary site.volcano-like ulcers[Histological examination is critical as the most valuable way to differentiate metastatic from primary tumors involving the gastrointestinal tract. The differences in the pathogenensis of metastatic or primary SCC in the gastrointestinal tract are also helpful to the differential diagnosis. The gastrointestinal tract may be metastatically involved by direct invasion, intraperitoneal dissemination and/or lymphatic or hematogenous cancer spread. The pathogenesis of SCC in the stomach and duodenum has not been well-elucidated. Several theories regarding the origin of SCC in the stomach and duodenum have been proposed, including nests of ectopic squamous cells, the proliferation of uncommitted mucosal basal cells into squamous cells, squamous metaplasia secondary to chronic mucosal damage, squamous differentiation in a preexisting adenocarcinonma and multipotent stem cells in the gastrointestinal mucosa ,10,11. Eke ulcers,12. The ke ulcers. UnfortuPatients with gastrointestinal metastases are often asymptomatic. Less frequently, these metastases can cause various symptoms such as gastrointestinal perforation, obstruction and/or hemorrhage. These nonspecific symptoms may be misinterpreted as indefinite complaints or as a side effect of treatment. For our patient, his complaint of dysphagia was transiently misdiagnosed as radiation-related esophagitis. Therefore, preemptive endoscopic examinations and careful biopsies are recommended in symptomatic patients. The presence of misplaced squamous cell nests or squamous metaplasia in the mucosa adjacent to the infiltrative tumor growth is highly suggestive of a primary SCC. However, in most of the reported cases of primary gastric SCC, precursor lesions were not demonstrated .et al. adopted CK5, p63 and p16 as immunohistochemical markers to confirm the diagnosis of SCC of the cervix metastatic to the duodenum [et al. studied the expression profiles of a series human epithelial cancers and their metastases by microarray technology and concluded that SCCs do not reflect their primary tissue expression profile [et al. performed immunohistochemical staining to compare the expression pattern of the epithelial-mesenchymal transition markers between primary SCC of the hypopharynx and a metastatic lesion of the duodenum. Impressively, dissociation of E-cadherin in cellular junctions and nuclear expression of Snail were detected in the primary tumor sample; in contrast, restoration of membranous E-cadherin and disappearance of nuclear Snail expression were demonstrated in the duodenal metastatic sample [Immunohistochemistry may also be useful in reaching the correct diagnosis. Over the past decade, expression levels of CK7, CK20 and TTF-1 have been widely used to distinguish pulmonary from gastrointestinal carcinomas, especially when evaluated as a panel of markers . Howeverduodenum . Strikinc sample . Their rThe prevalence of SCC of the stomach and/or duodenum is very low. The clinician should be aware of the possibilities of either metastatic or primary involvement of the stomach and duodenum with SCC. Distinguishing between the two requires extensive evaluation, including the patient’s clinical history, histological examination, immunohistochemical staining and possibly microarray data.Written informed consent was obtained from a son of the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.The authors declare that they have no competing interests.XS and JH designed the study. JH and YZ drafted the manuscript. All authors contributed to the intellectual content and approved the final version of the manuscript for publication. XS is the guarantor.


Lodoicea maldivica with the common name of coco de mer is used in Chinese medicine for treating cough. Native to Seychelles, Lodoicea maldivica seeds have commanded high prices for centuries due to its scarcity. This study aims to develop a molecular identification method for the authentication of Lodoicea maldivica seeds.The edible endosperm of Lodoicea maldivica respectively. DNA sequence of a sample was determined and compared with that of the Lodoicea maldivica reference material.DNA was extracted from the sample. Two polymerase chain reaction (PCR) systems were developed to amplify a region of the chloroplast DNA and the nuclear phosphoribulokinase (PRK) region specific to Lodoicea maldivica was successfully amplified and sequenced for identification.The PRK gene of Lodoicea maldivica seeds in fresh, frozen or dried forms was developed.A new molecular method for the identification of  Lodoicea maldivica (coco de mer), the sole member of the genus Lodoicea, is a fan-leaved palm native to Seychelles, bearing the largest and heaviest seed in the plant kingdom (weighing up to 30 kg). The seed is enclosed in a hard shell resembling a pair of coconuts joined in the middle. The seed is sometimes also referred to as the sea coconut, bum seed, double coconut, coco fesse or Seychelles nut. Lodoicea maldivica palms take between 25-50 years to reach maturity and bear fruit. The fruit may take one to three years to germinate . We compared the nucleotide sequence in pair between the reference material and each of the unknown samples using the bl2seq tool [Cocos nucifera with 100% homology in GenBank database [GenBank: HQ265608]  and reported by Lewis et al. in 2002 [Lodoicea maldivica reference material sequence obtained in this study showed a 99% homology to that of the [GenBank: AF453357] (Figure Lodoicea maldivica reference species used in this study was obtained from the country of origin of the species with official certificate. The present study provides an alternative source of nucleotide sequence data for GenBank for the identification of Lodoicea maldivica. The applicability and accuracy of the method have been demonstrated by the consistent results obtained from replicated analysis of the samples.All the amplified PRK genes were successfully sequenced. The nucleotide sequence of the seq tool . For tho] Figure . Current in 2002 . However in 2002  result oLodoicea maldivica seeds in fresh, frozen or dried forms was developed.A new molecular method for the identification of CITES: Convention on International Trade in Endangered Species of Wild Fauna and Flora; CTAB: hexadecyl trimethylammonium bromide; DMSO: Dimethyl sulfoxide; EDTA: Ethylenediaminetetraacetic acid; NCBI: National Center for Biotechnology Information; PCR: polymerase chain reaction; PRK: phosphoribulokinase; PVP: polyvinylpolypyrrolidone.The authors declare that they have no competing interests.CYM conceived and designed the study, collected the samples, performed the laboratory work, analyzed and interpreted the data, and drafted the manuscript. CSM supervised the project and finalized the manuscript. Both authors read and approved the final version of the manuscript.This article does not necessarily reflect the views of the Government of the HKSAR. The mentions of trade names or commercial products do not constitute any endorsement or recommendations.


This setup provides a possibility to address in depth the selected mechanisms that affect neuronal growth. The collected structural data (through staining and microscopy) and electrophysiological data (using microelectrode arrays) facilitate validation of computational models. Neuronal growth in dissociated cultures has been examined in several studies in order to access the role of activity in network development  or t or tin velopment . In addielopment ,9]. The. Thein velopment .The analyzed model consists of two types on neurons, most commonly observed in the neocortical cultures, the pyramidal cells and the nonpyramidal GABAergic cells, placed in a dish-like space with the density of cells corresponding to the experimental values. The phenomenological model that takes into account growth of every neurite is constructed using the description from the literature ,8]. It . It 8].


We explore the large spatial variation in the relationship between population density and burned area, using continental-scale Geographically Weighted Regression (GWR) based on 13 years of satellite-derived burned area maps from the global fire emissions database (GFED) and the human population density from the gridded population of the world (GPW 2005). Significant relationships are observed over 51.5% of the global land area, and the area affected varies from continent to continent: population density has a significant impact on fire over most of Asia and Africa but is important in explaining fire over < 22% of Europe and Australia. Increasing population density is associated with both increased and decreased in fire. The nature of the relationship depends on land-use: increasing population density is associated with increased burned are in rangelands but with decreased burned area in croplands. Overall, the relationship between population density and burned area is non-monotonic: burned area initially increases with population density and then decreases when population density exceeds a threshold. These thresholds vary regionally. Our study contributes to improved understanding of how human activities relate to burned area, and should contribute to a better estimate of atmospheric emissions from biomass burning. Fire is a natural process that has played a key role in the maintenance of natural ecosystems for millions of years, and regulates plant and animal population dynamics -3. HowevGuyette et al. (2002)  identifiThe objective of this study is to investigate the influence of population density on burnt area by exploring its spatial variability using Geographically Weighted Regression, and try to detect existence of critical thresholds in population density for fire behaviour using quantile regression. We then interpret the findings in the light of differences in major land use management classes.http://www.globalfiredata.org/. This spatial resolution can reveal first-order global and continental-scale patterns in burnt area .,20.19,202I)  sUnderstanding the complexity of the relationships between people and fire is important in a modelling context. Fire-enabled dynamic vegetation models can be used to predict the consequences of projected changes in climate on fire regimes see e.g. -46. Howe


Prostatic stones are a common condition in older men in industrialized countries. However, aging appears not to be the unique pathogenesis of these calcifications. Our morpho-constitutional investigation of 23 stone samples suggested that infection has a significant role in the lithogenic process of prostate calcifications, even without detection of infection by clinical investigation. Most stones (83%) showed bacterial imprints and/or chemical composition, suggestive of a long-term infection process. Chronic infection may induce persistent inflammation of the tissue and secondarily, a cancerization process within a few years. Thus, the discovery of prostate calcifications by computerized tomodensitometry, for example, might warrant further investigation and management to search for chronic infection of the prostate gland. Prostate cancer is the second most frequent cause of mortality due to cancer in males in the United States We noted the large chemical diversity of these prostatic stones. Carbonated calcium phosphate apatite  seems to be the major component, but several investigations show the presence of other mineral phases such as calcium oxalate monohydrate and dihydrate A previous investigation involving stone culture revealed that infected calculi in the prostate were implicated in relapsing urinary tract infection We investigated 23 prostatic stones obtained from the Saint-Louis and Tenon hospitals in Paris. The calcifications were collected from the prostate after radical prostatectomy or transuretral resection. The mean age of the patients was 71 years (range 35 to 87 years). All participants gave their verbal consent for use of the material. Samples were examined without knowledge of the name of the patient or other identifying data. Ethical approval for the study was obtained from the ethics committee of Tenon Hospital.−1, with resolution 4 cm−1 and time acquisition 1 sec/spectrum. The analysis was as previously described The FTIR spectroscopy was performed at Tenon Hospital. Each sample was analysed in absorbance mode on a Bruker Vector 22 spectrometer by accumulation of 32 spectra between 4000 and 400 cmEach prostatic stone was observed by Field-effect SEM (Zeiss SUPRA55-VP with an Everhart-Thornley secondary electron detector). To maintain sample integrity, each measurement was taken at low voltage (≤2 keV). Stones were imaged at similar magnification for comparison.Representative SEM images of normal stones and those with bacterial imprints (size about 500 nm) on the surface and in the core of stones are in Our SEM study revealed a high occurrence of bacterial imprints (78%) in 23 prostatic stones, which reveals a past or present infection of the prostate tissue; however, urinary tract infection was detected in only 6 (26%) cases. The large difference between number of reported infections and markers of infection within stones implies that aging may not be the only cause of prostatic calcifications. Infection and a lithogenic process induced by infection may play a role in most of the 99% of surgically removed prostate-containing stones.Staphylococcus aureus. Nevertheless, these imprints were not seen on all stone surfaces. Similarly to kidney stones, the bacteria can imprint on a particle surface such as CA or ACCP but not other crystal types such as whitlockite, octacalcium phosphate or brushite. Indeed, the size of CA and ACCP crystals is smaller than that of other phases, such as struvite The bacterial imprints had a specific spherical shape, which suggests infection by cocci germs. More precisely, the grape-like clustering, shape and size are common with staphylococci infection We discuss only the major phases. As previously reported, the main compounds of prostatic stones are calcium phosphate The deleterious consequences of chronic infection and inflammation have been well described in the cellular model The clinical interest of this paper is to draw attention to the high occurrence of asymptomatic infection of the prostate.History of urinary tract infection and risk of renal cell carcinoma have been found to be related. As reported by Parker et al. Prostatic stones are often considered to have no clinical significance, but the use of SEM showed for the first time the high frequency of bacterial imprints in these stones. Moreover, our data underline the specific chemistry of calcium phosphate phases, particularly the preponderance of whitlockite and ACCP in these calcifications. These results demonstrate the high occurence of bacterial infections in the prostate, often without any clinical symptoms.Inflammation induced by an infection may lead to cancerization of the tissue. Early detection of prostatic calcifications or stones could suggest a search for asymptomatic chronic infection. If an infection is detected, medical management and antibiotic treatment could avoid chronic inflammation of the tissue and further deleterious consequences. Thus, we suggest that discovery of prostate calcifications by imaging such as computerized tomodensitometry might warrant further investigations and management to search for chronic infection of the prostate gland.


We describe the saliva microbiome diversity in Batwa Pygmies, a former hunter-gatherer group from Uganda, using next-generation sequencing of partial 16S rRNA sequences. Microbial community diversity in the Batwa is significantly higher than in agricultural groups from Sierra Leone and the Democratic Republic of Congo. We found 40 microbial genera in the Batwa, which have previously not been described in the human oral cavity. The distinctive composition of the salvia microbiome of the Batwa may have been influenced by their recent different lifestyle and diet. The Batwa Pygmies, also known as Twa, are believed to be the original inhabitants of the equatorial forests of the Great Lakes region of Central Africa Traditionally, the Batwa have been semi-nomadic hunter-gatherers. However, relatively recent human activities, such as clearing of the forests for agriculture and creation of conservation areas, have pushed the Batwa from their traditional homeland and as a consequence their lifstyle has changed dramatically. Most Batwa now live on the borders of forest and agricultural areas but still use the forest on a daily basis. Batwa groups are small, rarely exceeding 50 people, and are often based around members of a particular clan.An interersting feature of the Batwa is that they differ significantly from neighboring Bantu agriculturalists in having fewer caries lesions and reduced tooth loss It is also possible that the oral microbiome of the Batwa may either influence, or be influenced by, the lower prevalence of caries.To investigate this further, we analyze here the saliva microbiome diversity of the Batwa in comparison with agricultural groups from similar enviroments in Africa, in order to address the following questions: 1) how different is the Batwa saliva microbiome from that of African agriculturalists; and 2) is the low level of dental caries in the Batwa associated with particular microbial taxa?All participants gave written informed consent. The protocol was in accordance with the Helsinki Declaration, and was approved by the Ethics Commission of the University of Leipzig Medical Faculty.Saliva samples were collected from: 39 Batwa from the Mpungo, Mukongoro, Kitariro, Nyakatare, Bikuto commuinities of Buhoma, Uganda: 20 individuals from Kinshasa, Democratic Republic of Congo (DRC); and 13 individuals from Freetown, Sierra Leone (SL). . DNA wasWe amplified a region of the microbial 16S rRNA gene containing variable segments V1 and V2, which were previously shown to be more informative than other parts of the 16S rRNA gene in terms of the number of phylotypes detected The PCR products were processed for parallel-tagged sequencing on the Genome Sequencer FLX platform, as described previously Ribosomal Database Project II (RDPII) database http://rdp.cme.msu.edu/seqmatch/seqmatch_intro.jsp) and a threshold setting of 90%, to assign a genus to each sequence. Diversity statistics and apportionment of variation based on the frequency distribution of genera within and between individuals were calculated with Arlequin 3.1 http://www.uga.edu/~strata/software/index.html ). UniFrac analysis The initial sequence reads were filtered to remove artifactual sequence reads . The filtered sequences were then searched against the Comparative data on the salivia microbiome diversity in two agricultural groups, from the Democratic Republic of Congo (DRC) and Sierra Leone (SL), came from another study .A total of 29728 sequence reads were obtained from the Genome Sequencer FLX. After filtering and removing sequence reads less than 200 bp, 24358 sequences remained . SequencThe results of comparing these sequences to the RDPII database are provided in A list of the top fifteen most frequent microbial genera found in these groups and their distribution is shown in Given the overall differences in the genera detected among the three groups, how strongly correlated are the saliva microbiome compositions of different individuals within and between groups? To address this question, we calculated correlation coefficients for the distribution of genera detected between each pair of individuals, both within and between groups . In geneWe also carried out an analysis of molecular variance (AMOVA) in order to investigate how much of the total variation in the saliva microbiome is due to differences within vs. among individuals from each group. The results indicate that the SL and DRC groups have very similar apportionments of variation, but the Batwa have much less differentiation between individuals, with 16% of the variation between individuals, vs. 30–32% for the other two groups . Thus, tIn order to test whether the patterns observed in the PCA plot are also seen at the sequence level, and not just from inferred genera, we also carried out UniFrac analysis and multidimenional scaling (MDS) based on UniFrac distances. The resulting MDS plot is highly consistent with the patterns observed in the PCA plot .To gain further insights into the influence of lifestyle and diet on the human saliva microbiome, we compared the saliva microbiomes of Batwa Pygmies  with two farming groups, from Sierra Leone and the Democratic Republic of the Congo.We found significantly higher diversity in the saliva microbiome of the Batwa (127 different genera) compared with the groups from DRC (54 different genera) and SL (71 different genera). Rarefaction analysis  indicateCloacibacterium, found at low frequency in several Batwa, was discovered recently but in a different enviroment, namely in municipal wastewater We found 59 microbial genera that are unique to the Batwa . AlthougWe compared the list of microbial genera found in Batwa with the Human Oral Microbiome Database Although groups from SL and DRC are geographically distant , they sh2 by bacterial ureases which are produced by a particular subset of oral bacteria, including Actinomyces naeslundii and oral HaemophilusActinomyces does not vary significantly between the Batwa and the other two human groups, but for Haemophilus, the frequency of reads is significantly higher in the Batwa (0.034) than in the other two groups . Moreover, Haemophilus was identified in almost every Batwa individual Click here for additional data file.Table S1Count of detected microbial genera in Batwa Pygmy individuals.(XLS)Click here for additional data file.Table S2Count of sequence reads assigned to each detected microbial genus in Batwa and groups from the Democratic Republic of the Congo (DRC) and Sierra Leone (SL).(XLS)Click here for additional data file.


The mouse model of oxygen-induced retinopathy (OIR) has been widely used for studies of retinopathy of prematurity (ROP). This disorder, characterized by abnormal vascularization of the retina, tends to occur in low birth weight neonates after exposure to high supplemental oxygen. Currently, the incidence of ROP is increasing because of increased survival of these infants due to medical progress. However, little is known about changes in the chronic phase after ROP. Therefore, in this study, we examined morphological and functional changes in the retina using a chronic OIR model. Both the a- and b-waves in the OIR model recovered in a time-dependent manner at 4 weeks (w), 6 w, and 8 w, but the oscillatory potential (OP) amplitudes remained depressed following a return to normoxic conditions. Furthermore, decrease in the thicknesses of the inner plexiform layer (IPL) and inner nuclear layer (INL) at postnatal day (P) 17, 4 w, and 8 w and hyperpermeability of blood vessels were observed in conjunction with the decrease in the expression of claudin-5 and occludin at 8 w. The chronic OIR model revealed the following: (1) a decrease in OP amplitudes, (2) morphological abnormalities in the retinal cells (limited to the IPL and INL) and blood vessels, and (3) an increase in retinal vascular permeability via the impairment of the tight junction proteins. These findings suggest that the experimental animal model used in this study is suitable for elucidating the pathogenesis of ROP and may lead to the development of potential therapeutic agents for ROP treatment. Retinopathy of prematurity (ROP) is a disease that affects the retinas of premature infants. The key pathological change, namely, retinal neovascularization, is associated with local ischemia followed by the subsequent development of neovascularization. In the more severe forms of the disease, the abnormal vascular changes may progress to retinal detachment. Once retinal detachment occurs, the prognosis for recovery of good visual acuity is poor. As a result, ROP is the leading cause of preventable childhood retinal dysfunction. The World Health Organization's Vision 2020 programme targets ROP as an avoidable disease requiring early detection and treatment to prevent blindness The severity of ROP is graded in stages from 1 to 5. Blindness from ROP is caused by retinal detachment due to tractional retinal angiogenesis (stage 5). Recently, it was reported that even mild ROP (stages 1 or 2) can cause permanent functional sequelae The oxygen-induced retinopathy (OIR) model is widely used for studies of retinal neovascular diseases such as ROP and diabetic retinopathy The blood-retinal barrier (BRB) is a selective barrier of the eye and is composed of capillary blood vessels. The BRB plays an essential role in protecting the neural tissues from toxic materials and maintaining the visual and neural functions of the retina. BRB breakdown is one of the most important pathophysiological changes in ischemic retinal diseases such as ROP and diabetic retinopathy To create an effective therapeutic strategy for avoiding the blindness caused by ROP, it is essential to understand the relationship between retinal dysfunction and structure degeneration. Although there is evidence that abnormalities occur in both the retinal blood vessels and retinal cells in ROP The purpose of the present study is to elucidate the interaction of the retinal blood vessels and retinal function using a chronic OIR mouse model.To determine visual function after exposure to high oxygen, we performed ERG analyses. Similar to previous reports To investigate the damage in each retinal layer after exposure to high levels of oxygen during infancy, we evaluated the number of cells in the the ganglion cell layer (GCL) and the thicknesses of the inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), and outer nuclear layer (ONL). In the OIR mouse model retinas, the thicknesses of the IPL, OPL, and INL decreased compared to those in normal mouse retinas at 4 and 8 w . HoweverWe also examined retinal blood vessel morphology in OIR mice. As shown in The permeability of retinal blood vessels was evaluated by tracer experiments using FITC-dextran (2000 kDa) conjugated with Hoechst 33342 (616 Da). This tracer has previously been used to successfully evaluate the permeability of tight junctions of epithelial cells The main results of the present ROP study can be summarized as follows: (1) there were deficits in OPs, which are useful indices of IPL dysfunction; (2) retinal damage was found primarily in both the IPL and INL where many retinal blood vessels exist; (3) retinal vascular permeability increased via breakdown of the barrier properties in retinal blood vessels. These findings indicate that retinal hyperpermeability can cause retinal dysfunction and that the murine chronic OIR model is suitable for evaluating the pathogenesis of BRB disruption. Furthermore, these results demonstrate that the present model is useful as a chronic OIR model to clarify the relationship between visual function and retinal vasculature.In this study, ERG readings reflected the loss of retinal function. To assess the retinal function in normal and OIR mice, the amplitudes of a- and b-waves were measured as they are indicative of photoreceptor function and the functional integrity of the INL, respectively In addition to compromised ERGs, we observed significant decreases in the thicknesses of the IPL and INL in the OIR mice compared to those of normal mice with the recovery of a- and b-wave amplitudes  and 3. ONext, to verify retinal neurovascular units in the chronic OIR mice, we evaluated both retinal permeability and expression of retinal microvascular tight junction molecules. We found that retinal hyperpermeability was observed with BRB disruption in chronic OIR mice, and we evaluated claudin-5 and occludin expression, which are involved in the assembly of tight junctions between neural microvascular endothelial cells In conclusion, these findings in the murine chronic OIR model indicate that deficits in OPs may be a sign of BRB disruption, which may be indicative of the early symptoms of retinal vascular abnormalities. The chronic OIR model revealed vascular impairment dysfunction without neovascular tufts; therefore, it may be appropriate to investigate the pathogenesis of ROP that leads to blindness caused by neovascularization without retinal detachment. The abnormalities of the OPs seen on ERG can be used to diagnose BRB disruption, and appropriate treatments, such as protection of endothelial cells in the early phase, may be able to prevent the loss of vision observed in ROP.The experimental designs and all procedures were approved by the Animal Experimental Committee of Gifu Pharmaceutical University . All experiments were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research, and monitored by the Institutional Animal Care and Use Committee of Gifu Pharmaceutical University. C57BL/6 mice  were used in this study. The mice were kept under controlled lighting conditions (12∶12-h light/dark).The OIR mouse model was created as described previously ERG measurements in mice were performed as described in our previous report 2)]. According to a previous report 2)]. In the present study, OPs amplitudes were measured by using ERG with all frequency (0.3–500 Hz).To demonstrate the oscillatory potentials (OPs), the OP amplitudes were measured in the time between the a- and b-wave peaks; the factors were OP number , age , and flash intensity . A P value of <0.05 was considered statistically.Data are presented as the means ±standard error of the mean (S.E.M). The control mice participated in all experiments, and some OIR mice were used in multiple experiments. Statistical comparisons were made using Student's


Problems experienced within the first year of an infant's life can be precursors of later mental health conditions. The purpose of this study was to examine the frequency and continuity of difficult behaviors in infants at 3 and 6 months of age and the associations of these difficulties with biomedical and psychosocial factors.This study was a part of an ongoing prospective birth-cohort study. Study participants were 189 uniparous mothers and their full-term newborns. The index of infant difficult behavior was constructed. This index was then associated with the following factors: delivery mode, newborn function after birth, maternal emotional well-being, risk behavior, subjective evaluation of the quality of the relationship of the couple, and attitudes toward infant-rearing.Common difficult behaviors, including crying, sleeping and eating problems, were characteristic for 30.2% of 3 month old and for 22.2% of 6 month old full-term infants. The expression of infant difficult behaviors at the age of 3 months increased the likelihood of the expression of these difficulties at 6 months by more than 5 times. Factors including younger maternal age, poor prenatal and postnatal emotional well-being, prenatal alcohol consumption, low satisfaction with the couple's relationship before pregnancy, and deficiency of infant-centered maternal attitudes towards infant-rearing increased the likelihood of difficult behaviors in infants at the age of 3 months. Low maternal satisfaction with the relationship of the couple before pregnancy, negative emotional reactions of both parents toward pregnancy (as reported by the mother) and the deficiency of an infant-centered maternal attitude towards infant-rearing increased the likelihood of infant difficult behaviors continuing between the ages of 3 to 6 months. Perinatal biomedical conditions were not related to the difficult behaviors in infants.Our study suggests that early onset of difficult behavior highly increases the risk for the continuation of difficult behavior during infancy. In general, the impact of prenatal psychosocial environment on infant behavior decreases from the ages of 3 to 6 months; however, some prenatal and preconceptional psychosocial factors have direct associations with the continuity of difficult behaviors through the first half-year of an infant's life. Studies of mental health condition precursors underscore the importance of the infancy period for a child's long-term adjustment ,2. InfanSome studies indicate that early biomedical circumstances surrounding the birth of a child affect parental behavior, infant behavior, and later postnatal development. Indicators of a more stressful delivery are linked to more frequent crying and fussing in the infant, and to more difficulty regulating the infant's behavior . The modUnlike subjects in other research that focuses on infants of various risk groups, the subjects of this study were full-term newborns. To carry out an in-depth investigation of the roots of difficult behavior it is also important to clarify the mechanisms of such behavior, as well as the reasons behind it within normal development and conditions. The aims of our study are to examine the frequency and continuity of infant difficult behaviors at 3 and 6 months of age, and to determine the biomedical and psychosocial factors linked with early infant difficulties. In this paper, we focus on the common difficult behaviors manifested by infants, which include crying, sleeping, and eating problems.This study is the initial part of an ongoing prospective birth-cohort study started in 2009. The research was performed with the approval of the Kaunas Regional Biomedical Research Ethics Committee (No. P1-143/2007). The study participants were uniparous mothers who gave birth to full term newborns (≥ 37 weeks of gestation) in the Hospital of the Lithuanian University of Health Sciences, Kaunas Clinics. This is a Baby Friendly Hospital, where the medical staff strictly follows the principles of early breastfeeding initiation and rooming-in. After vaginal delivery, newborns stay on close skin-to-skin contact with their mothers. After caesarean section delivery, newborns stay in the nursery room until their mothers are transported to the intensive care ward, and within two hours, the newborn is transported to the mother for breastfeeding.This study consisted of three stages. The first stage was performed in the clinics, and included the collection of medical data about delivery and newborn function, as well as the collection of data from questionnaire surveys for mothers during their stay. Written informed consent was obtained from women who participated in the study together with their infants. The questionnaires were given to mothers during the second or third day after childbirth; the new mothers were asked to answer them at their own convenience. The first stage involved data received from 546 mothers. The second stage was performed three months postpartum, during which women were asked to complete mail-in questionnaires. A total of 242 participants responded to the request and completed the questionnaires. The third stage of research was carried out six months postpartum, during which women were once again asked to complete the mail-in questionnaires. Completed questionnaires were received from a total of 261 respondents. We selected the data of those women who participated in all three stages of the study, a total 189 participants, for the analysis. Their mean age was 28.38 (SD = ± 5.70).χ2 = 8.95, p = 0.011).We also checked whether the demographic data differed between women who participated in all three study stages, and were included in analysis (participants), and those who were excluded from analysis . The analysis indicated no differences according to mean age, place of residence, or marital status. However, a marked difference was discovered with regards to higher education (college or university degree) among participants: 138 participants (70.4%) had received higher education, and only 199 of the non-participants (57.8%) had received higher education , small for gestational age (SGA) neonates  and large for gestational age (LGA) neonates  according to Lithuanian national birth weight standards .Apgar scores 1 and 5 minutes after birth were used to evaluate newborn functioning.The neonatal neurological and adaptive capacity was assessed at 2 hours, and at 24 hours, after birth by a neonatologist using the Neurological and Adaptive Capacity Scale (NACS) [e (NACS) . The NACnd-3rd day postpartum, psychosocial data about prenatal and short postpartum period were collected. The participants completed the Prenatal environment questionnaire, whose questions covered demographics, pregnancy planning, emotional reactions toward the conception, the relationship with the husband or partner, tobacco use and alcohol consumption, and emotional experiences during pregnancy and postpartum. All prenatal variables were scored retrospectively during this stage.During the first study stage, on the 2demographic characteristics of the participants were highlighted using questions about maternal age, education, and family structure.The pregnancy planning. Next, the emotional reactions toward pregnancy were evaluated by the questions 'Which statement best reflects your reaction toward pregnancy?' and 'Which statement best reflects your husband or partner's reaction toward your pregnancy?' with possible answers 'happy', 'conflicting feelings', 'upset', or 'other'. In the analysis, the answers were divided into two groups: the first group as the positive reaction, and the second group as the ambivalent or negative reactions, including conflicting feelings, upset feelings, or other negative emotional reactions.The participants were asked whether the pregnancy was planned or not-planned in order to evaluate The quality of the couple's relationship was subjectively evaluated by mothers using the Likert type scale, rating from 'very bad relationship' (1 point) to 'very good relationship' (5 points). Mothers were asked to evaluate the relationship with husband or partner before and during pregnancy.Tobacco and alcohol use during pregnancy was evaluated by the questions 'Did you smoke cigarettes during pregnancy?' and 'Did you consume alcohol during pregnancy?' The possible answers were as follows: 'not at all,' 'several times during the whole pregnancy,' 'once or several times a month,' 'once or several times a week,' and 'everyday.' The women who reported not smoking during pregnancy were categorized as 'nonsmokers', those who reported smoking several times during whole pregnancy, or several times a week or a month were defined as 'non-regular smokers'; those who had smoked everyday were categorized as 'regular smokers'. According to alcohol consumption, women were divided into two groups: those who reported prenatal alcohol consumption and those who did not report alcohol consumption during pregnancy.stressful and traumatic experience during pregnancy, women were asked whether or not they had experienced any stressful and traumatic events during pregnancy.To evaluate Negative emotional experiences during pregnancy were evaluated by questions on how often during pregnancy they were experienced emotions such as irritability/bad temper, feeling low and feeling nervous; mothers were given the possibility to choose one of five statements for each emotion: 'almost daily,' 'more often than once a week,' 'almost every week,' 'almost every month' and 'rare or never,' evaluated from 1 to 5 points. The total sum score was used to evaluate negative emotional experience, where lower sum score indicates more often experienced negative emotional state.Depressiveness 2 to 3 days postpartum was measured by the Edinburgh postnatal depression scale [on scale . The Liton scale . Cronbacnd stage of the study, the participants completed the Infant development and social environment questionnaire. This survey asked questions regarding parental social support, the relationship of the couple, maternal emotional state, breastfeeding, postpartum maternal attachment, and maternal attitudes toward child rearing. Women were also asked if the father of the infant is living with her and the child, and if he helps with childcare.Three months postpartum, during the 2The quality of the couple's relationship, stressful and traumatic experiences, as well as negative emotional experience during three months postpartum, and depressiveness 3 months postpartum were measured using the same questions as during the 1st stage.Breastfeeding the baby was also included in the analysis. Women were asked if they are breastfeeding their infants or not.Maternal attitudes toward caring for an infant were assessed using the Infant-rearing attitudes and beliefs scale [very strong agreement to very strong disagreement, how much they endorse the viewpoint expressed in each of the eight statements. The statistical analysis showed that Cronbach's alpha for this measure was 0.74.fs scale . The meaWomen's perception of infant's difficult behaviors scale [Infant difficult behaviors were selected as a dependent psychosocial variable in this study. It was assessed at the ages of 3 and 6 months using rs scale . The oriχ2 test and Z test, where appropriate, to assess the differences in the prevalence of characteristics between different groups of respondents. A measure of agreement (kappa) was used to assess the consistency of responses on an infant's problem behavior between the ages of 3 and 6 months. In testing statistical hypotheses, a p-value of less than 0.05 was considered significant.Basic and advanced statistics procedures of the Statistical Package for Social Sciences (SPSS) for Windows version 15.0 software package were used to conduct data analysis. The first analyses included descriptive statistics and primarily frequencies; this provided an understanding of the distributions of the respondents' demographic, social, psychological and medical characteristics during the pregnancy and childbirth period, as well as infant's health, mental development, and behavioral problems. We applied the Women's perception of infant's difficult behaviors scale were combined into one derivative variable, labeled "Index of Infant's Difficult Behaviors" (IIDB). It was not the simple sum of scores in responses to each of 7 questions of the scale, but rather a linear combination of them with different weights. The values of weights in this linear combination were estimated from the total data set by the SPSS Factor Analysis procedure requesting a single factor, and then that single factor (IIDB) for each record  was calculated and saved. The method of factors extraction was based on principal component analysis [In order to assess infant difficult behavior in general, 7 items of the analysis . The weiThe last analyses included binary univariate logistic regression analysis procedures. Odds ratios (OR) and 95% confidence intervals (CI) by adjusting data for infant's gender were calculated in order to analyze the associations between different demographic, social, psychological and medical variables and IIDB. For this purpose, the sample was split into two groups according to low or high rates of infant's difficult behaviors (negative or positive IIDB) at the infant's ages of 3 months, 6 months and at both 3 and 6 months. Calculations were performed separately for each of the three sample groupings.The statistical analysis showed that the different items of Index of Infant Difficult Behaviors at 3 and 6 months varied according their weights. The strongest weights at 3 and 6 months included items such as infant crying Table . We thusThe Index of Infant Difficult Behaviors illustrated that a high rate of difficult behaviors was characteristic for 30.2% of 3 month old and for 22.2% of 6 month old full-term infants. 13.2% of infants had a high rate of difficult behaviors at ages of 3 and 6 months. According to the data presented in Table Logistic regression analysis proved that a strong association exists between difficult behaviors at 3 and 6 months: the likelihood of difficult behaviors at the age of 6 months increased more than five times  if such problems were reported at the age of 3 months. The prevalence of a high problem rate among 6-month-old infants was greater if these infants were characterized as having a greater amount of problems at the age of 3 months, compared with 3-month-old infants with a low problem rate (43.9% and 12.9% respectively). However, a remarkable decrease of problem behavior after 3 months was also observed; greater than half (56.1%) of the infants with problem behavior at 3 months stopped by 6 months of age  toward conception, low satisfaction with the couple's relationship before pregnancy, and rigid parent-centered maternal attitudes towards infant rearing. This suggests a strong significance of parent-related and family-related characteristics for an infant's development. Young families, especially those with poor relationships, low readiness for parenthood, and deficient child-oriented attitudes should therefore receive early prevention programs, which focus on teaching infant-rearing skills.In general, it can be stated that the direct significance of some aspects of prenatal history  decrease during the first half year of infancy, while aspects of the postnatal child environment  become more significant. This could be explained by the complex interplay between a child and his or her environment over time, and the confounding effect of risk factors. Bidirectional effects of the child and of the environment are highlighted in transactional models  and are The results of this study also document the importance of maternal prenatal stress and emotional well-being in the emergence of infant difficult behaviors at 3 months of age, although it was not related with later infant problems. The effects of prenatal anxiety/stress on a child's difficulties developing in cognitive, behavioral, and emotional ways are documented in numerous studies . The preOur study endorsed the importance of the paternal role in a child's development from the very beginning of pregnancy. Based on the study data, the quality of a couple's relationship before pregnancy and paternal emotional reactions toward conception could have long-lasting effects on infant behaviors and may be of greater significance than the quality of the couple's relationship later in time. Despite growing research interest and recognition of the importance of the father in a child's development, data on the influence of the fathers' role during the early stages of pregnancy and infancy are still lacking. Little attention is given to the relationships of couples before pregnancy; numerous studies exist about the effect of a couple's relationship on offspring development after childbirth or during pregnancy. It is proposed that marital conflict has an impact on the emotional arousal of children and the regulation of emotion and behavior within children . Data haThe results revealed the significance of both maternal and paternal emotional response towards conception in respect of infant difficult behaviors. Negative emotional response to the fact that mother became pregnant increases the likelihood of continuation of infant difficult behaviors by about 4 times. Some earlier studies indicate that infants whose conception was unintended by their father are at an elevated risk for adverse health events . FathersThe study indicated that the continuation of infant difficult behaviors between the ages of 3 and 6 months is highly related to postpartum maternal attitudes toward infant rearing. Infants of mothers expressing more parent-centered and rigid attitudes towards infant-rearing had nearly three times of a greater risk for continual behavioural difficulties in comparison with mothers who expressed more flexible and infant-centered attitudes. Other studies provide evidence that mothers' self-reported attitudes correspond with their child rearing behaviors . The parHowever, we are considering that factors such as the emotional reactions towards conception and attitudes towards infant rearing are interrelated; perhaps they can be seen as a reflection of the parental acceptance of the child and preparedness to integrate him into the family system. Lack of emotional acceptance and preparedness at this stage might have a later effect on attachment to the child. Studies inspired by attachment theory highlight that the feelings and attitudes individuals have about parent-child relationships, even before they become parents, are predictive of the subsequent infant-mother attachment pattern and the emotional quality of their future parenting behaviors . HoweverWomen's perception of infant's difficult behaviors scale should also be noted as another methodological weakness of the present study. Thus, further empirical evaluations are needed.Several limitations need to be addressed regarding the present study. Our decision to follow the development of full-term newborns (≥ 37 gestation weeks) could be mentioned not only as an advantage, but also a limitations of our study. Since our study was conducted only among low risk infants, the sample size was limited. The second limitation of the study was that all prenatal measures were collected post-natally during the first days after childbirth. The special emotional state of a recently delivered woman could influence responses to the questionnaire. In our study, all information about independent variables  and the dependent variable (infant difficult behaviors) were collected from mothers' reports. It was assumed that mothers acted as primary caregivers of infants, predisposing them to greater access to information about infant behavior than other family members. However, the fact that mothers were the only informants in our study may poorly reflect the real circumstances regarding infant behavior and family life; this is another important limitation of this study. Factors such as maternal personality and emotional state after childbirth could significantly influence the reports. This limited the possibility of obtaining more objective data. It is also important to note that information about the fathers' emotional reactions towards pregnancy, as well as the evaluation of the couple's relationship, was also obtained from mothers and could reflect more of the woman's evaluations about her husband/partner than the real feelings and emotions of the fathers. The results of our study, indicating important role of fathers' emotions and the parents' relationship quality on infant difficult behaviors, shows that these issues should be analyzed more comprehensively in further research, with an emphasis on collecting reports from fathers as well. Relatively low reliability scores using the Lithuanian version of the Our research revealed that early manifestation of infant difficult behaviors increases the risk for the continuity of these problems during the first half-year of infancy. In general, the impact of prenatal psychosocial environment on infant difficult behaviors decreases between the ages of 3 and 6 months. Although, some prenatal and preconception factors, such as a poor relationship between the couple and negative emotional reactions towards conception, are directly associated with the continuity of infant difficult behaviors throughout the infant's first half year of life. The established importance of attitudes towards child rearing suggests that parental education could be effective in various prevention or intervention programs.The authors declare that they have no competing interests.EM and RJ initiated the study. RJ, EM, NZ, GS participated in the design of the study. GS, NZ and AZ participated in the construction of the current data analyses, data management and interpretations: AZ performed the statistical analysis of the data; GS prepared the draft of the current manuscript, and NZ provided important contributions to the manuscript as well. All of the authors were involved in the revision of the manuscript and contributed to the interpretation of the data. All authors read and approved the final manuscriptThe pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2431/12/44/prepub


The aim of this study was to determine the diagnostic accuracy of routine Papanicolaou stain (PAP) and Silver stained Nucleolar Organizer Regions (AgNOR) staining in brush biopsies taken from suspected oral lesions for early detection of oral cancer.Brush biopsies were collected from macroscopically suspicious lesions of the oral cavity of 34 patients and 10 normal-aged and sex-matched controls. The numbers of AgNORs were counted in 100 squamous epithelial cell nuclei per slide after silver staining of the smears (Ploton’s one-step method).Sensitivity and specificity of PAP analysis in the oral smears for detection of oral cancer and normal cells was 91.176% and 100%. The positive and negative prediction values were 100% and 76.92%, respectively. Sensitivity and specificity of AgNOR analysis in the oral smears for detection of oral cancer and normal cells was 100%. The positive and negative prediction values were 100% each.Based on the above facts, we conclude that brush biopsy in conjunction with AgNOR staining is an easily practicable, non-invasive, safe and accurate screening method for the detection of macroscopically suspicious oral cancerous lesions. Because of its simple technique and high reliability for cellular proliferation, AgNOR staining in brush smears can be used as an adjunct to other routine cytological diagnoses for the early detection of oral cancer. However, further investigations with more number of study samples will be needed to establish this correlation beyond doubt. Oral squamous cell carcinomas (OSCCs) currently hold the sixth position in the worldwide cancer statistics, with a dExfoliative cytology is an easy, non-invasive procedure and hence could be carried out even on slightest suspicion regarding the nature of the given lesion. AlthoughIn the last few years, AgNOR analysis is being frequently used to determine the prognosis of many malignant lesions. NORs canThe purpose of this study was to determine the diagnostic accuracy of routinely performed PAP staining as compared with AgNOR staining in brush biopsies of suspected oral lesions.This study was carried out in the Department of Oral Pathology and Microbiology, Government Dental College and Hospital, Aurangabad. The study population consisted of 44 subjects (including control), from which 88 smears (two smears per subject) were obtained. After thorough evaluation, 10 subjects for the control group were selected from age- and sex-matched subjects with prior consent and these subjects comprised the group I category of this study.The 34 subjects having clinically diagnosed or suspicious of cancerous lesions (excluding recurrent lesions or those who had taken some sort of treatment) were grouped separately and comprised the group II category of this study. The brush biopsies were obtained and diagnosed before scalpel biopsies clarified the nature of the oral lesions histologically. The quantification of AgNOR counts was performed blindly without the knowledge of the cytological or histopathological report.After thorough clinical examination and consent, the subjects were subjected to 5-min gargling and the lesional areas were wiped off of excessive saliva and surface debris using a moistened gauze piece. Lesional areas with erythematous patches were usually preferred as collection sites. In case of highly keratotic or exophytic lesions, fissured or ulcerative areas were preferred for collecting the cells.Two cytologic smears were obtained from the pathologic area in question using a cytobrush plus GT . The head of the cytobrush cell collector was moistened with water and was then firmly held against the mucosa of the lesional area. Then, gentle pressure was applied to the brush until the bristles curled or tiny bleeding spots were evident. In this position, the brush was rolled over the lesional site and was rotated for 10 full turns. The cytobrush cell collector was then rolled on glass slides by applying a continuous motion from one end of the slide to the other.The spray-fixed smears were stained by a commercially available RAPID-PAP Papanicolaou stain kit .Evaluation of Papanicolaou the-stained smears was carried out according to the standardized procedure at a magnification of ×450. The cells suspected to be abnormal were evaluated at higher magnifications and the location of abnormal cells was marked on the cover slip by an ink dot .et al. T28 T2628]27et al.et al. stained the smears only for 20 min while in the present study, the staining time used was 55 min. Because of an increase in the staining time in the present study, the small NORs present in the given nucleus may have fused with each other due to continued deposition of silver for a longer time. As a result, the small-sized dot-like precipitations merge and the discrimination between small individual dots may not be possible.Remmerbach In the present study, the diagnostic accuracy of routine PAP method and AgNOR staining were compared for the diagnosis of OSCCs. The routine PAP-stained smears were graded into five grades based on Papanicolaou’s criteria. Thus, thet al. reported the false-negative rate of exfoliative cytology to be ranging from 0% to 29% and, in his own study, the rate was 31%.[The false-negative rate of PAP staining for diagnosing OSCCs in the present study was 23.08%. Folsom  was 31%. The falset al.[Scuibba and Remmet al. have repFurthermore, epithelial dysplasias and borderline lesions represent morphological alterations that are suspicious for malignancy, but do not provide sufficient evidence for its definitive diagnosis. This mayOn the other hand, the latter situation offers the opportunity for adjuvant methods to identify the malignancy earlier than subjective interpretation of histological or cytological images. Here lies the importance of AgNOR analysis, which, due to its simplicity, has an edge over immunohistochemical methods for the similar purpose. When AgNOR staining was employed in the present study, the sensitivity and specificity of AgNOR staining was 100%. The negative and predictive values were also 100%. Here lies the advantage of AgNOR analysis because, unlike the PAP method, this analysis was able to not only diagnose all cases of verrucous carcinomas but also differentiate them from OSCCs. Hence, NOR analysis may be useful as a quantitative marker of incipient cellular alterations even before the histological hallmarks of changes can be detected.Based on the above facts, AgNOR analysis appears to be far superior to routine PAP staining when used for the detection of OSCCs in exfoliated cells.All smears contained more than sufficient number of cells required for microscopic analysis and from all cell layers. This proves the efficiency of the cytobrush cell collector. The mAgNOR per nucleus is a reliable marker of neoplastic squamous cells in oral smears. This method is able to increase the sensitivity for the detection of malignant and specificity for benign cells in oral smears and, thus, decreases the rate of cytologically false-negative or positive diagnoses. The AgNOR technique in exfoliative cytology can be used as an adjuvant diagnostic aid to routine cyto- and histopathology for differentiating between benign and malignant lesions of the oral cavity.Although there seems to be a generally positive correlation between AgNOR counts and degree of malignancy, further investigations with more number of study samples will be needed to establish this correlation beyond doubt. However, their ease of demonstration and high specificity to cellular proliferation makes them the best available cytopathological marker in the arsenal of the oral pathologist.


Marine microorganisms are rich source for natural products which play important roles in pharmaceutical industry. Over the past decade, genome-based studies of marine microorganisms have unveiled the tremendous diversity of the producers of natural products and also contributed to the efficiency of harness the strain diversity and chemical diversity, as well as the genetic diversity of marine microorganisms for the rapid discovery and generation of new natural products. In the meantime, genomic information retrieved from marine symbiotic microorganisms can also be employed for the discovery of new medical molecules from yet-unculturable microorganisms. In this paper, the recent progress in the genomic research of marine microorganisms is reviewed; new tools of genome mining as well as the advance in the activation of orphan pathways and metagenomic studies are summarized. Genome-based research of marine microorganisms will maximize the biodiscovery process and solve the problems of supply and sustainability of drug molecules for medical treatments. Symploca sp. VP642 from Palau, and the synthesis of plinabulin (NPI-2358) was inspired by halimide and phenylahistin, the former of which was isolated from a marine fungus Aspergillus sp. CNC-139. The structures of other interesting compounds with novel structures or potent activities can be retrieved from the recent literatures and review papers [The marine environment covers more than 70% of the Earth's surface and has been proven to be a rich source for both biological and chemical diversity. Marine natural products have become fascinating targets for biologists and chemists for discovery of lead compounds for clinical development for the past five decades –10. Marin NPI-235 was inspw papers . Although natural products embrace a wide range of entities such as biomaterials and biocatalysts as well as biocontrol agents other than drug molecules, due to space limitation, in this paper, most of the cited natural products focused upon are compounds with the potential to be developed as drugs and used in medical treatments. Taking the advantage of established microbial cultivation technology, the promising natural products identified from marine microorganisms provide great potential to solve the supply and sustainability problem of useful drugs to cure human diseases. High-throughput screening (HTS) of drug molecules demands abundant diversity of the compound libraries to get more hits for drug discovery. To combat the emergence of drug-resistant pathogens, as well as to improve the efficacy and safety of medical treatment, new drug leads are urgently needed. The key point of improving the success of natural product screening and development is to improve the diversity and the size of natural product library for new drug discovery. In this paper, the recent progress in the genomic research for natural compound discovery is summarized, with the focus on the important roles of genetic diversity deduced from genome mining on new drug discovery. http://www.genomesonline.org), of which 1674 complete genomes are registered. And in the mean time, 204 archaeal, 6087 bacterial, and 2003 eukaryal genome projects are ongoing, which are double of those reported in 2009 [Nocardiopsis sp. strain TFS65-07 and marine-derived fungus strain Aspergillus sp. MF297-2 were reported to be sequenced, and studies on the biosynthesis of thiopeptide antibiotic TP-1161 and prenylated indole alkaloids, stephacidin and notoamide metabolites, were subsequently guided by the bioinformatics analysis of gene clusters located in these genomes [Genomic sequencing of microorganisms in the recent years has unveiled unprecedented insights into the biosynthetic potential of microbial cells, and thus the discovery of natural products has entered into the new postgenomic era. The genomic sequencing data of various living organisms have been accumulating rapidly in recent years. According to the Genomes On Line Database (GOLD), there are 5831 genome sequencing projects registered in GOLD, while according to the last update , 10298 genome projects are documented currently in GOLD  dedicated to polyketide synthase (PKS) and/or nonribosomal peptide synthetase (NRPS) biosynthesis, which are megasynthases that are responsible for many active natural product biosynthesis and are especially popular tools for combinational biosynthesis [ S. tropica has great potential to produce novel natural products and that the biosynthetic potential of this species, like that of many other actinomycetes that were genomically sequenced, is considerably greater than that has been observed by cultivation and chemical analysis [The genome sequence of the first seawater-requiring marine actinomycete, reported . Bioinfoectively . The genynthesis –35. The analysis , which pAlthough natural products from marine microorganisms have already widened the spectrum of chemical diversity, increasing the diversity of natural compounds is still the prerequisite for HTS to get enough new hits for drug discovery. And in the meantime, the classical activity-based screening is increasingly challenged by repeated discovery of known compounds, as well as the limitation of available assay methods. Therefore, new techniques are needed to maximize the diversity of natural products for the discovery of new therapeutic agents, not only to increase the efficacy and decrease the toxicity, but also to combat the emergence of drug resistance. The development of genomic sequencing techniques as well as the bioinformatics tools in recent years has profound influence on the discovery of new drugs. Nowadays it is much easier to have the genome of the interested organisms sequenced, especially for relatively small genomes of microorganisms. The availability of genomic sequences and the subsequent genome mining of microorganisms have provided new insights in the biosynthetic potential of the cells, and thus the studies of marine natural products have entered into the new postgenomic era. There is an increasing trend to expand the studies on the chemical diversity of natural products from bioassay-based screening to genetic diversity studies, from the studies of natural diversity to creation and utilization of recombinant diversity , and in the meantime, it is well accepted that the chemical diversity and genetic diversity not only exist in different microbial species, but also exist in the different strains in one species . TherefoIt has been observed that the genetic elements responsible for the biosynthesis of many, if not all, secondary metabolites are clustered in the bacterial or fungal genomes to form gene clusters, which include the genes encoding biosynthetic enzymes, as well as genes responsible for regulation and resistance . This geStreptomyces sp. S187 was obtained, and bioinformatics analysis guided the identification of the gene cluster responsible for a new glycopeptide antibiotic (unpublished data). Genome scanning is another successful approach, in which the random genome sequence tags (GSTs) prepared from the genomic DNA library are screened using degenerate primers to identify the gene cluster, and the products of the gene cluster can be subsequently searched using various genomics-guided strategies [In the recent years, more and more studies have focused on the “from genetics-to-chemistry” route for natural product research, that is, to use gene-based screening strategy to study the biosynthetic potential of the microbial strains, followed by molecular cloning experiments and chemical purifications , 39. A grategies , 41, whiAlthough the cost of genome sequencing is declining, it is still not affordable for every lab for natural product research. Two questions may arise concerning the whole genomic sequencing. Why do we need to know all the sequences on the genome? What can we do next after obtaining the genomic sequence? S. coelicolor. To answer the first question, it is well accepted that the genetic loci in the genome can be roughly grouped by genes involved in primary metabolism and genes involved in secondary metabolism. Because not all genes are expressed in all the conditions, the chemical analysis of the cultivation broth under certain conditions cannot fully explore the biosynthetic potential of the cells. Furthermore, not all genes can be fished out by gene-based screening using degenerate PCR; therefore, sequencing the genome can fully access the genetic loci responsible for natural product biosynthesis. At the same time, genetic loci involved in primary metabolism are also tightly linked to the biosynthesis of natural product by providing precursors and cofactors ; therefoThe answer to the second question relies on the rapid development of tools for genome mining, which were presented in several excellent reviews elsewhere –51. SeveS. tropica strain CNB-440 is one good example [slm gene cluster codes for polyene macrolactam polyketide, and investigation of the fermentation broth using characteristic UV chromophores of polyene units led to the identification of structural fragments of salinilactam A series compounds. Inspection of the structure fragments suggested that salinilactam A was derived from a PKS with at least 10 extension modules. This information was useful to help resolve and properly assemble the repetitive DNA sequences associated with the slm PKS, which was used in combination of partial NMR-based structural fragments to accurately predict the gross chemical structure of salinilactam A. What is more, bioinformatics analysis also assisted in the correction of the initial assignment of the C-28 methyl group complicated by overlapping olefinic NMR signals. The salinilactam A structure was later verified by comprehensive NMR analyses of the purified natural product to confirm the bioinformatics-based total structure assignment.As indicated above, the polyketides and nonribosomal peptides are synthesized by modular PKS or NRPS system, and extensive knowledge has been accumulated on the relationship between the structures of these compounds and the organization of the multienzymes. Successful mining of ribosomally synthesized peptides has also been reported and summarized . Bioinfo example . InitialS. pacifica” with that of S. tropica [S. pacifica”, which is related to the 41 kb gene cluster in S. tropica for salinosporamide A biosynthesis, but the genes coding for the enzymes in the chloroethylmalonyl-CoA pathway are absent in “S. pacifica.” This information guided the isolation of salinosporamide K, which structurally resembles salinosporamide A [In another recent report, salinosporamide K was discovered by comparative genomic analysis of “ tropica . A truncramide A .The structures of salinilactam A and salinosporamide K described above were presented in S. coelicolor is the early example in which this approach was employed [Pseudoalteromonas tunicata D2 was overexpressed in E. coli, which led to the isolation of two new metabolites, 3-formyl-L-tyrosine-L-threonine dipeptide and 3-formyl-L-tyrosine [Biosynthetic gene clusters that are related to the production of yet-unknown metabolites are often referred to as “orphan” . The bioemployed . Alternaemployed . In a retyrosine . This woAspergillus nidulans. Overexpression of the putative pathway-specific regulatory gene located in the silent gene cluster resulted in the identification of the novel hybrid PKS-NRPS product aspyridone [The failure of producing certain natural products can be attributed to the poor expression or silence of the biosynthetic gene cluster. Therefore, overexpression of the regulatory genes, either pathway-specific regulatory genes or global regulatory genes, can lead to the production of novel compounds. One example is the discovery of aspyridones in pyridone . AlternaS. coelicolor after deleting a putative pathway-specific regulatory gene (scbR2) that encodes gamma-butyrolactone receptor protein [On the other hand, some regulatory genes can also exert inhibitory effect on the production of certain compounds; in this case, the deletion of such regulatory genes readily results in the activation of silent pathways and production of novel compounds. In a recent report, a new yellow-pigmented secondary metabolite was observed in  protein . This stOther strategies for genome mining including genomisotopic studies and in vitro reconstitution of biosynthetic enzymes were described in reviews elsewhere , 59. It E. coli and Streptomyces lividans. Clone libraries are produced independently of the host cultivation or fermentation and then subjected to sequence-based screening or function-based screening. This cultivation-independent approach is termed metagenomics. During the past decade, many successful metagenomic studies have been reported. Onnamides and theopederins are antitumor polyketides produced by an uncultured Pseudomonas sp. symbiont of marine sponge Theonella swinhoei. Biosynthesis genes of onnamides and theopederins were isolated from the metagenome of the marine sponge, and bioinformatics analysis of the biosynthetic genes strongly indicated a prokaryotic origin, suggesting that these potent antitumor agents may be produced by the symbiotic bacteria [Candidatus Endobugula sertula from the marine bryozoan [Psammocinia aff. Bulbosa [Marine symbiotic microorganisms are rich source of active natural products and are thought in some cases to be the true producers of potent drug candidates instead of the marine invertebrate including sponges, tunicates, and bryozoans . Shotgunbacteria . An entibryozoan . In anot Bulbosa . Combini Bulbosa , active The structures of some compounds studied by metagenomic analysis were presented in Exploration of the diversity of marine natural products is greatly promoted by various genome-based strategies described in this paper, and new chemical structures and novel bioactivities unveiled by these methods are complementing the traditional bioassay-guided studies. However, as indicated by Glöckner and Joint , the sucBacillus strain NK2018 [Poribacteria [On the other hands new tools in the detection of natural products are continuously being developed to explore the chemical diversity of microbial natural products. The imaging mass spectrometry (IMS) tools were used to visualize the spatial distribution of secondary metabolites produced by marine cyanobacteria , providin NK2018 . Single-bacteria , and thiThe exploration of the diversity of marine microorganisms using the combined classical activity-based screening with the genome mining in both the culturable and unculturable microorganisms is presented in Recent years have witnessed the exponentially increasing genome sequencing information, and genome mining analyses have revealed tremendous capability of marine microorganisms to produce active natural products as therapeutic agents. Studies on the biosynthetic machinery and regulatory mechanisms of natural product biosynthesis will greatly maximize the diversity exploration of marine natural products. Nowadays the diversity studies of natural products have been shift from pure chemical analysis to the combination of genetic manipulations and chemical synthesis, and the diversity of natural products can be further enlarged by genetic engineering of biosynthetic genes. Genomic research of marine microorganisms will have great impact on the discovery of novel natural products for medical treatments and will contribute to efficient drug discovery from marine environment.


Enterobacteriaceae (ESBL) in neonatal unit associated with an alarming mortality rate.A surveillance of multiresistant bacteria was conducted from April to October 2012 in 2 pilot hospitals in Dakar. Results revealed higher infection rates caused by extended-spectrum beta-lactamase producing The objective of this study was to evaluate the management of infectious risk in the neonatal unit.An extensive investigation was carried out. Following indicators were evaluated: existence and functionality of the infection control committee (ICC), availability of resources for hand hygiene, knowledge and perception of healthcare workers (HCW) on hand hygiene , invasive procedures care and presence of ESBL in the patient environment (by swabbing).The ICC was set up but was not functional . Alcohol-based handrub and disposable towels were rarely found in the unit. More than 80% of HCW didn’t know the hand hygiene indications and thought that handwashing was more effective against germs than handrubbing. Mechanical ventilation and parental nutrition were used in more than 80% of neonates with ESBL infection. The environmental analysis revealed the presence of ESBL strains on surfaces, beds and antiseptic vials.Our investigations indicated the potential factors that could explain the high rates of ESBL infection observed in the neonatal unit. The ICC should be functional because of its central role in the management of infectious risk. Material resources  should be provided in the unit to ensure good hand hygiene practices. Educational programmes on hand hygiene indications and technics will be implemented for all HCW. Guidelines for the cleaning, disinfection and decontamination must be produced. Moreover, an evaluation of practices should be planned regularly to ensure adequate practices.None declared


Employing a neuron-specific retrograde gene-transfer vector (NeuRet vector), we have recently developed a novel technique that achieves pathway-selective ablation in the primate brain. This technique is mediated by immunotoxin (IT) and eliminates a neuronal population that constitutes a particular pathway, leaving other pathways intact. By means of this technique, we have made an attempt to remove the hyperdirect pathway selectively from basal ganglia circuitry. The hyperdirect pathway links the motor cortex to the subthalamic nucleus (STN) directly and plays a crucial role in motor control. After electrical stimulation in the motor cortex, triphasic responses consisting of an early excitation, an inhibition, and a late excitation are usually elicited in the internal pallidal segment (GPi). Several pieces of pharmacophysiological evidence imply that the early excitation may be derived from the hyperdirect pathway. In our experiments, the NeuRet vector expressing human interleukin-2 receptor α-subunit was injected into the STN of macaque monkeys. Then, IT injections were performed into the supplementary motor area (SMA). When single neuron activity in the GPi was recorded in response to the SMA stimulation, it was found that the early excitation was significantly reduced with neither the inhibition nor the late excitation affected. The spontaneous firing rate and pattern of GPi neurons remained to be altered. This clearly indicates that IT-mediated tract targeting successfully eliminated the hyperdirect pathway with spontaneous activity of STN neurons unaffected. The electrophysiological findings were histologically confirmed by retrograde and anterograde neuronal labeling. The overall data define that the motor cortically driven early excitation in GPi neurons is conveyed through the hyperdirect pathway. The IT-mediated pathway-selective ablation technique will provide a powerful tool for elucidating information processing in various neural networks. To know about a variety of higher brain functions systematically, it is essential to elucidate the architecture of complex and elaborate neural networks. For clarifying the functional role of a given pathway, it is an effective way to explore behavioral and physiological changes due to ablation of a neuronal population that constitutes the target pathway. Neuronal targeting mediated by immunotoxin (IT) has been established in mice as a genetic method for eliminating a specific neuron group from a certain neural network . RecentlFigure 1; Figure 1; In our recent work, we have applied the IT-mediated pathway-selective elimination technique to the primate brain, because the use of non-human primates as animal models is critical for investigating higher brain functions. Employing the nigrostriatal dopamine pathway as a test system, we have first established the basic methodology with a neuron-specific retrograde gene-transfer vector (NeuRet vector) that has newly been developed with improved neuron specificity . Next, aFigure 2A). To verify the capability of the NeuRet vector for efficient retrograde gene transfer into the nigrostriatal pathway, we injected the vector encoding the green fluorescent protein (GFP) transgene into the striatum (caudate nucleus and putamen) of crab-eating monkeys. Intrastriatal injection of the NeuRet vector produced a large number of GFP-positive neurons in the nigra . These neurons were immunostained for tyrosine hydroxylase, a key enzyme for dopamine biosynthesis , indicating the transgene expression in the nigrostriatal dopamine neurons. Moreover, we assessed the extent of gene transfer with the NeuRet vector around the injection sites in the monkey striatum. The vector displayed a low level of gene transfer into neuronal cells, and the level of vector transfer into glial cells was also quite low in the striatum . Therefore, the NeuRet vector mediates enhanced retrograde gene transfer into neuronal cells, whereas it reduces the efficiency of gene transfer into glial cells around the injection sites.We have developed a new vector system that permits NeuRet by pseudotyping the HIV-1-based lentiviral vector with fusion glycoprotein C type (FuG-C) consisting of a hybrid of rabies virus glycoprotein (RV-G) and vesicular stomatitis virus glycoprotein VSV-G; . InteresFigure 1; The NeuRet vector expressing IL-2Rα was injected into electrophysiologically identified sites in the STN of Japanese monkeys. Following the vector injections into the STN, activity of GPi neurons was recorded in response to electrical stimulation in the supplementary motor area (SMA). In most of the GPi neurons in which certain responses were induced, we observed a triphasic response pattern consisting of an early excitation, a subsequent inhibition, and a late excitation, as previously reported in normal monkeys . Compared with a control condition (before the IT injections), the amplitude of the early excitation was largely  reduced after the IT injections. On the other hand, the amplitude of the inhibition and the late excitation remained relatively unchanged, although the late excitation slightly decreased with no significant change . In addition, virtually no alterations were found in the latency of the inhibition or the late excitation, or the duration of the inhibition or the late excitation. In the control monkey in which no vector injections were made into the STN, the response pattern of GPi neurons on SMA stimulation was essentially the same before and after IT injections into the SMA . Thus, the IT injections into the SMA combined with the injections of the NeuRet vector expressing IL-2Rα into the STN abolished the cortically evoked early excitation in the GPi without affecting either the inhibition or the late excitation.After IT injections into the SMA, especially its arm region, neuronal activity in the GPi was recorded in response to the SMA stimulation. Many of the recorded GPi neurons exhibited a biphasic pattern, an inhibition followed by a late excitation without an early excitation . Neurons in the GPi fired randomly at high frequency before the IT injections, and no apparent changes were observed after the IT injections . These results suggested that the firing rate and pattern of GPi neurons remained unchanged even after the removal of the cortico-STN projection.Furthermore, the spontaneous firing rate and pattern were compared before and after the IT injections into the SMA. The spontaneous firing rate was left intact . Moreover, immunostaining for NeuN revealed that the IT injections into the SMA caused no marked tissue damage . After the BDA injections into the SMA forelimb region, anterogradely labeled axon terminals were so largely decreased in the STN, as compared to the control case (data not shown). In remarkable contrast, dense terminal labeling from the SMA was seen in the striatum, especially the putamen, as in the control case (data not shown). These anatomical data clearly indicated that cortico-STN projection originating from the SMA arm region was selectively eliminated without affecting either the corticostriatal projection or the cortico-STN projections from SMA regions with other representations than the arm.In the monkeys subjected to the disappearance of the early excitation responding to the SMA stimulation, we performed retrograde and anterograde neuronal labeling by injecting Fluoro-ruby (FR) into the STN and biotinylated dextran amine (BDA) into the SMA. After the FR injection into the STN, retrogradely labeled neurons in the SMA were much fewer in the arm region where the IT injections were primarily aimed than in the face and leg regions  Blo neurons .For selective removal of the hyperdirect pathway, the NeuRet vector expressing IL-2Rα was injected into the STN, and, subsequently, IT was injected into the SMA in our experimental protocol. Our histological examination clearly indicated that cortical neurons in the arm region of the SMA projecting to the STN were selectively ablated. In such model monkeys, GPi neuron activity was recorded in response to electrical stimulation in the SMA. The SMA stimulation yields selective activation of SMA-recipient zones in the basal ganglia . It was According to the cortically driven triphasic response pattern elicited in GPi neurons, the hyperdirect pathway conveys excitatory signals from the motor cortex toward the GPi, bypassing the striatum, with shorter conduction time than signals via the striatum that arise from both the direct and the indirect pathways see , 2002a. Since no explicit motor abnormalities were seen in our experimental conditions, changes in the activity of GPi neurons in response to the cortical stimulation following a limited amount of elimination of the hyperdirect pathway may not be enough to elicit behavioral alterations. Accordingly, there is a need to determine the relationship between the extent of the elimination of the selected pathway and the expression of altered behaviors. The IT-mediated tract targeting achieves selective ablation of a given pathway in primates. This novel technique will provide a potent strategy to explore not only specific functional roles of individual pathways constituting a particular neural network, but also large-scale operative mechanisms underlying the entire network.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.


Ileal adenomas associated with familial adenomatous polyposis are a common finding. Many recent studies following panproctocolectomy for familial adenomatous polyposis have confirmed the presence of multiple ileal adenomas and an increase in ileal mucosal proliferation. In this study, we present a case of invasive adenocarcinoma arising in a severely dysplastic tubulovillous adenoma in the ileostomy of a patient with familial adenomatous polyposis; also, we present a literature review. To the best of our knowledge, only very few cases have been reported in the literature.A 59-year-old Caucasian woman developed a primary adenocarcinoma in her ileostomy, complicating the stoma 31 years after its formation.Primary adenocarcinoma following panproctocolectomy for familial adenomatous polyposis is a very rare clinical entity. The risk of developing adenocarcinoma in those patients increases with time. Patient education and medical examination of the stoma are of paramount importance and should be implemented early with the need of designing a surveillance protocol for early detection and management of ileal adenomas, especially in longstanding stomas. Ileal adenomas associated with familial adenomatous polyposis (FAP) are a common finding. Many recent studies following panproctocolectomy for FAP have confirmed the presence of multiple ileal adenomas and an increase in ileal mucosal proliferation. The management protocol for FAP is prophylactic colectomy with either restorative proctocolectomy with formation of ileal pouch reservoir or ileorectostomy. Panproctocolectomy and terminal ileostomy were the first-line management option in 1950s. Currently, this procedure is performed only for cases with recurrent rectal or ileal pouch adenocarcinoma. In this study, we present a case of adenocarcinoma in the ileostomy of a patient with FAP; in this case, invasive adenocarcinoma arising in a severely dysplastic tubulovillous adenoma was found. Also, we present a literature review. To the best of our knowledge, only 11 cases in English and one in German [A 59-year-old Caucasian woman with a previously diagnosed FAP had total colectomy with ileorectal anastomosis 34 years ago followed three years later by resection of the rectal stump and fashioning of an end ileostomy in her left iliac fossa; neither the operative details nor the pathology reports were available. Her medical history shows that she has multiple medical comorbidities, including severe sero-negative rheumatoid arthritis, lumbar disc herniation, total abdominal hysterectomy and bilateral salpingo-oophorectomy., osteoarthritis in both hips, eczema, chronic obstructive pulmonary disease, renal calculi, and chronic renal disease stage 3.Three years ago, she was referred to the colorectal clinic because of abnormal fungating growth at the stoma Figure . She wasen bloc small bowel resection with the ileostomy and safety margin of the skin around the stoma. About 45 cm of adherent small bowel was removed alone with the ileostomy specimen. The rest of the small bowel was examined and did not show any gross pathology. A new stoma was created in her right iliac fossa. She went home seven days after a smooth recovery from the operation.A computed tomography scan of her chest, abdomen, and pelvis after oral and intravenous contrast showed no evidence of metastatic disease. Carcinoembryonic antigen was within normal range. After a discussion in the colorectal multi-disciplinary team meeting, a decision was made to keep the duodenal polyps under surveillance and to carry out a barium meal and follow through study. The results of the contrast study were clear; therefore, our patient underwent a laparotomy with Pathology showed an invasive, moderately differentiated adenocarcinoma arising in a severely dysplastic tubulovillous adenoma with foci of high-grade dysplasia at the mucocutaneous border Figure . The tumAs discovered at a follow-up one year ago, she had developed a polypoid mass at the ileostomy site Figure . The resFAP is an autosomal dominant inherited disease characterized by the presence of hundreds of adenomatous polyps in the colon and rectum. The annual reported incidence ranges from one in 5000 to one in 17,000 live births . The incMany factors have been implicated in the malignant transformation of the ileostomy mucosa: colonic metaplasia and dysplasia of ileal mucosa, chronic irritation due to either physical or chemical trauma, ileitis, and development of chronic inflammatory polyps and malignant transformation in a pre-existing benign adenoma. In this case, we believe that the invasive adenocarcinoma was secondary to malignant transformation in a pre-existing adenoma as evident by the pathology report.en bloc resection of the terminal ileum with its mesentery, surrounding stoma skin, and relocation of a new ileostomy. Our patient was followed up to 18 months without evidence of malignant recurrence. However, she developed a benign adenomatous polyp, which was locally excised with refashioning of the stoma.Table en bloc resection of the ileum with a generous margin of the surrounding skin and relocation of the stoma.The number of reported cases for primary adenocarcinoma following panproctocolectomy for FAP is small. Nevertheless, it is expected that the numbers will increase in longstanding ileostomies in parallel with the duration of the stoma. Patient education and medical examination of the stoma should be implemented early with the need of designing a surveillance protocol for early detection and management of ileal adenomas, especially in longstanding stomas. Surgery should include an EGD: esophagogastroduodenoscopy; FAP: familial adenomatous polyposis.Written informed consent was obtained from the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.The authors declare that they have no competing interests.AH performed literature searches, wrote the paper, and assisted in the operation. RT contributed in writing the amendments and in revising the manuscript. PJM and SG performed the operation and supervised the final revision of the paper. All authors read and approved the final manuscript.