Gastric tube after esophagectomy can be the site of local recurrence or the development of second primary tumor which implies poor prognosis. The study presents an extremely rare case of a patient after Ivor-Lewis esophagectomy for squamous cell carcinoma, in whom there was detected local recurrence in the anastomosis associated with metachronous primary tumor in gastric tube. Esophageal reresection with the upper part of the stomach was performed. Left colonic segment supplied by middle colic vessels transposed through retrosternal route was used as new esophageal substitute. Esophagectomy with lymph node dissections combined with adjuvant therapy remains the basic method of esophageal cancer surgical treatment. The stomach is the most commonly used esophageal substitute after esophagectomy. Owing to the availability of new, more precise diagnostic techniques and to the improvement of the late outcomes of esophageal cancer treatment, there increases the detectability of the cases of second primary malignancies in the stomach . The coeThe surgical treatment of a cancer diagnosed in the gastric tube after esophagectomy is a difficult and rarely undertaken surgical challenge associated with the risk for severe complications and high mortality.A female patient, MZ, aged 61 years, referred primarily for surgical treatment due to esophageal cancer detected in the epicardial region. Endoscopic examination revealed a primary tumor located 30 cm from the incisors. Endoscopic evaluation of the stomach lumen was impossible due to esophageal stricture at the tumor level. Histopathological examination of tumor specimens led to the diagnosis of squamous cell carcinoma. Preoperative thoracic computed tomography (CT) detected neither tumor infiltration on the surrounding tissues nor the enlargement of the regional lymph nodes in the thorax and abdominal cavity.Based on the results of diagnostic investigations which confirmed the possibility of local excision of the tumor, the patient was qualified for surgical treatment.3N0M0, Stage II A). Microscopically esophageal and gastric rings were free from neoplastic cells. The proximal resection margin was 4 cm. Eighteen lymph nodes of the following groups were removed: upper thoracic paraesophageal lymph nodes (2), right thoracic paratracheal lymph nodes (1), bifurcation lymph nodes (2), middle thoracic paraesophageal lymph nodes (2), right pulmonary hilar lymph nodes (1), lower thoracic paraesophageal lymph nodes (2), diaphragmatic lymph nodes (1), posterior mediastinal lymph nodes (2), lesser curvature lymph nodes (3), left gastric artery lymph nodes (1), and common hepatic artery lymph nodes (1). All lymph nodes were free of cancer cells. The patient’s postoperative course was uncomplicated and she was discharged from hospital after 17 days. After surgery, the patient did not report for further treatment at an oncological center.Partial Ivor-Lewis esophago-gastrectomy was performed with conventional two-field en block lymphadenectomy using a gastric tube as an esophageal substitute. The esophagogastroplasty was performed in the right pleural cavity. The postoperative histopathological examination showed squamous cell carcinoma keratodes G-2 invading the adventitia without lymph node metastasis (pTThe follow-up endoscopy performed 6 months after esophagectomy showed: (A) mucosa at the site of anastomosis: uneven, congested with superficial lesions covered with fibrin, unchanged esophageal mucous membrane proximal to anastomosis. About 3 cm below the anastomosis was a stocky polyp of 10 to 12 mm in diameter (B) surrounded by normal gastric mucosa. Congested mucosa without lesions were in the gastric tube distal segment. The results of histopathological examinations revealed: (A) gastric mucosa segment with the features of foveolar hyperplasia, (B) polyp from the gastric tube - squamous cell carcinoma keratodes . The result of CT after esophagectomy was . En bloc dissection of residual esophagus was performed to the level of the apex of the chest and then the esophageal stump was cut 8 cm above the anastomosis Figure . The esopT2N0Mx (primary focus), the tumor sounded by normal gastric mucosa; and (c) lymph nodes from the thoracic area (7) and neck (4) without tumor cells.The results of postoperative histopathological examination were (a) distal esophageal stump with anastomosis: recurrence of squamous cell carcinoma keratodes G-2, tumor emboli in lymphatic vessels, proximal esophageal stump free of tumor infiltration; (b) tumor located in gastric tube, squamous cell carcinoma Eight weeks after surgery, in the course of the second cycle of chemotherapy, dysphagia developed. Endoscopy revealed no anastomotic stenosis. Within a 3-week period an effective endoscopy-guided dilatation with a balloon was performed twice. Six months after reoperation no recurrence of the neoplastic process was observed in follow-up diagnostic tests.The use of the stomach as an esophageal substitute after esophagectomy is commonly accepted by the majority of surgeons ,5. The sIn the case of metachronous tumors located in the gastric tube the therapeutic management depends on the depth of tumor infiltration to the gastric wall -12. In tet al.[Gastric tube resection requires reconstruction of gastrointestinal tract with the useof colonic or small intestine conduit. In such situations colon graft is preferred owing to its proper length, reliable blood supply, and fewer complications ,12,15. Iet al.. The nonGastric tube resection with the creation of a new esophageal substitute from the colon is a difficult but possible treatment option in selected cases of recurrence or second primary cancer of the stomach detected in postoperative follow-up..Written informed consent was obtained from the patient for publication of this report and any accompanying imagesThe authors declare that they have no competing interests in this paper.SJ composed the case report, prepared and edited this manuscript, contributed it conception, collected the data and conducted a literature search. ŁP and MW participated in the data collection and gave final approval for this version of the manuscript. All authors read and approved the final manuscript.


Nilaparvata lugens  and Laodelphax striatellus  are two of the most important pests of rice. Up to now, there was only one mitochondrial genome of rice planthopper has been sequenced and very few dependable information of mitochondria could be used for research on population genetics, phylogeographics and phylogenetic evolution of these pests. To get more valuable information from the mitochondria, we sequenced the complete mitochondrial genomes of BPH and SBPH. These two planthoppers were infected with two different functional Wolbachia (intracellular endosymbiont) strains (wLug and wStri). Since both mitochondria and Wolbachia are transmitted by cytoplasmic inheritance and it was difficult to separate them when purified the Wolbachia particles, concomitantly sequencing the genome of Wolbachia using next generation sequencing method, we also got nearly complete mitochondrial genome sequences of these two rice planthoppers. After gap closing, we present high quality and reliable complete mitochondrial genomes of these two planthoppers.N. lugens (BPH) and L. striatellus (SBPH) are 17, 619 bp and 16, 431 bp long with A + T contents of 76.95% and 77.17%, respectively. Both species have typical circular mitochondrial genomes that encode the complete set of 37 genes which are usually found in metazoans. However, the BPH mitogenome also possesses two additional copies of the trnC gene. In both mitochondrial genomes, the lengths of the atp8 gene were conspicuously shorter than that of all other known insect mitochondrial genomes . That two rearrangement regions (trnC-trnW and nad6-trnP-trnT) of mitochondrial genomes differing from other known insect were found in these two distantly related planthoppers revealed that the gene order of mitochondria might be conservative in Delphacidae. The large non-coding fragment (the A+T-rich region) putatively corresponding responsible for the control of replication and transcription of mitochondria contained a variable number of tandem repeats (VNTRs) block in different natural individuals of these two planthoppers. Comparison with a previously sequenced individual of SBPH revealed that the mitochondrial genetic variation within a species exists not only in the sequence and secondary structure of genes, but also in the gene order (the different location of trnH gene).The mitogenomes of The mitochondrial genome arrangement pattern found in planthoppers was involved in rearrangements of both tRNA genes and protein-coding genes (PCGs). Different species from different genera of Delphacidae possessing the same mitochondrial gene rearrangement suggests that gene rearrangements of mitochondrial genome probably occurred before the differentiation of this family. After comparatively analyzing the gene order of different species of Hemiptera, we propose that except for some specific taxonomical group (e.g. the whiteflies) the gene order might have diversified in family level of this order. The VNTRs detected in the control region might provide additional genetic markers for studying population genetics, individual difference and phylogeographics of planthoppers. The mitochondrial genome is the most commonly used molecular marker for phylogenetic studies, population genetics and dynamics, phylogeography  and evenThe order Hemiptera is the largest group of the hemimetabolous insects  and inclNilaparvata lugens and the small brown planthopper (SBPH), Laodelphax striatellus (Hemiptera: Delphacidae) are two of the most important pests in rice fields of many Asian countries. By transmitting virus diseases [Wolbachia[Wolbachia strains, we also obtained nearly the complete mitochondrial genomes of BPH and SBPH. The Wolbachia strain wStri infecting SBPH can induce complete Cytoplasmic Incompatibility (CI), a reproductive modification that typically results in embryonic lethality between crosses of infected males and uninfected females. On the other hand, the Wolbachia strain wLug infecting BPH cannot induce CI at all.The brown planthopper (BPH), diseases  and causdiseases . These tWolbachia which caWolbachia,24. WhiltrnH gene) were verified using natural populations. In addition, comparing mitochondrial genome of SBPH sequenced in this study with a previously sequenced individual revealed a remarkable variability between these two individuals.After gap-closing, the complete mitochondrial genomes of these two most important rice piercing and sucking type pests, BPH and SBPH were determined. We analyzed the nucleotide composition, codon usage and genomic rearrangement of these two mitochondrial genomes and compared them with mitochondrial genomes of other hemipteran species. Some special characteristics of mitochondria of these two planthoppers  in ‘A + T’ rich regions; location of N. lugens (BPH) mtDNA and another scaffold containing 3 contigs of L. striatellus (SBPH) were identified in the assemblage of the wLug and wStri genome  which were reported so far. The unusual length of the mitochondrial genome in N. lugens is partly due to a long putative control region and a long repeat region containing three repeats of trnC gene  of BPH, trnS2 (UCN) of SBPH which harbor a simple loop in the dihydrouridine (DHU) arm and trnH of SBPH could have a loop-stem structure like trnT of BPH  that forms a loop is found in many Hemiptera [trnS1 (AGN) has a completed loop-stem structure while that of trnS2 (UCN) has a loop. In contrast, the trnS1 (AGN) has a loop and trnS2 (UCN) has a completed loop-stem structure in previously reported SBPH [trnH gene of SBPH which formed a weak loop-stem structure of the T-arm (TΨC) in this study possessed a stable loop-stem structural of the T-arm (TΨC) in previous study [All of the tRNA genes have the typical cloverleaf secondary structure, except for emiptera ,18,27-30emiptera . For SBPted SBPH . Furtherus study .trnS2 (UCN) (SBPH) and 57 bp for trnS1 (AGN) (BPH) to 71 bp for trnK (BPH and SBPH) and trnV (BPH)  and rrnL . They are located between trnL1 (CUN) and an A+T-rich region, separated by trnV.The length of all tRNA genes ranged from 56 bp for trnC-trnW and nad6-trnP-trnT) associated with tRNAs and PCGs were found in both of BPH and SBPH . But the most striking feature was the different locations of the trnH gene in two individuals of SBPH. The trnH gene is located between nad5 and nad4 genes in this study just as in all other hemipteran species sequenced so far, but Song and Liang (2009) previously reported that this tRNA gene is located between nad4L and nad6 genes . This feature is common in hemipteran insects [The mtDNAs of BPH and SBPH contain the full set of PCGs usually presented in animal mtDNA. Except for the  Insects   were the shortest in insects known so far. In fact, the longest atp8 gene (228 bp) was found in Bemisia tabaci (Hemiptera: Aleyrodidae) [Pachypsylla venusta) [Lymantria dispar) [atp8 gene had a length of about 160 bp. Taking into account the length of atp8 gene of planthoppers discovered in this study, it seems that the length distribution of atp8 gene in Hemiptera was wider (From 99 bp to 228 bp) than that of all other orders of insects reported so far. The sequence alignment of atp8 gene with other insects revealed that nearly 60 bp length of sequence in the 3′end of this gene was lost in these two planthoppers. Does the short length of this gene change the structure of its encoded protein? Does length change of this gene alter the functions or efficiency of this gene? These aspects are worth further studying.It is worth mentioning that the lengths of rodidae) . In almovenusta)  to 183 b dispar) . In mostThe abundance of codon families and Relative Synonymous Codon Usage (RSCU)  in PCGs rrnS and trnI-trnQ-trnM gene cluster  in the A+T-rich region. This poly “T” structure might be conservative in Delphacidae and be responsible for the control of replication and transcription of mitochondria.Neuroctenus parus) to 86.3% (Aleurodicus dugesii). These two extreme A+T content species were inferring gene arrangement [The composition of the majority strand (J-strand) of SBPH mtDNA is A = 7099 (43.2%), T = 5581 33.97%), G = 1527 (9.29%) and C = 2224 (13.54%). The composition of the J strand of BPH mtDNA is A = 7393 (41.96%), T = 6165 (34.99%), G = 1661 (9.43%) and C = 2400 (13.62%). Similarly to most hemipteran insects, the nucleotide compositions of the two planthopper mitogenomes are significantly biased toward A and T . Comparing with 48 mitochondrial genomes of hemipteran species, we found that there is considerable variation in base composition among different hemipteran species: ranging from 68.9%  to 0.2765 (Lycorma delicatula) with the BPH and SBPH mtDNA exhibiting a slight different AT-skew (0.0909 for BPH and 0.119171 for SBPH). The GC-skews of hemipteran species vary from −0.2827 (Acyrthosiphon pisum) to 0.2086 (Neomaskellia andropogonis) with the BPH and SBPH mtDNAs exhibiting very similar GC-skew (−0.1826 for BPH and −0.1842 for SBPH). Similarly, the AT/GC-skew values exhibit different ranges among three suborders: AT-skew: Heteroptera, 0.0642 - 0.2181, Sternorrhyncha, -0.1812 - 0.0956, Auchenorrhyncha, 0.0623 -T 0.2765; GC-skew: Heteroptera, -0.2656 -(−0.0965), Sternorrhyncha, -0.2810 - 0.2086, Auchenorrhyncha, -0.2827 - (−0.0783).The hemipteran AT-skews vary from −0.1812  of Sternorrhyncha, all species of Aleyrodidae exhibit converse AT/GC-skew values Figure  as descrrrnL gene and rrnS gene with the entire mitochondrial genome, PCGs show more significant positive correlation (R2 = 0.9674) than rrnL gene (R2 = 0.8873) and rrnS gene (R2 = 0.7189) , complete trnH gene (62 bp), two intergenic nucleotides and a portion of nad4 gene (96 bp). All sequenced individuals  have an identical sequence of the trnH gene which was the same with that of the mitochondrial genome of SBPH sequenced in this study. Considering the low probability of many different sites in the trnH gene related sequences caused by PCR or sequencing, we still cannot exclude the possibility that some sporadic individuals of SBPH possessing the same location of trnH gene as previous study exist in some special regions in China. Anyway, according to the mutation sites  of all sequences of the region compassing trnH gene, 16 haplotypes of the sequence were identified  were observed in natural populations  and one ATP synthase gene (atp8). Because of the different locations of trnH gene, positions of the start codon of nad5 and nad6 genes and the end codon of nad4 and nad6 genes had been changed. Comparing the region (from nad5 to nad6) of mitochondrial genomes of SBPH with that of previous study, we found the differences were 57 bp deletion in the front of nad5 gene, 21 bp deletion at the end of nad4 gene and 84 bp insert in the front and 55 bp deletion at the end of nad6 gene. Furthermore, there also existed 21 bp absent in the nad2 gene and 60 bp absent at the end of atp8 gene  of mitochondrial genome of these two SBPH, the other eight PCGs and two ribosomal RNA genes represented significant difference in the evolution rates  showed the fastest mutation rate among these genes. The mutation rates of the other PCGs ranged from 0.0022 to 0.0076. The evolution rate of rrnS gene (0.8%) was five times faster than rrnL gene (0.16%). In general, except the trnH and two trnS genes, the other tRNA genes were conserved (only few of them have single variant site).Excluding five length distinct genes  detected in the control region might provide additional genetic markers for studying population genetics, individual difference and phylogeographics of planthoppers. The comparative analyses of two sequenced mitochondrial genomes of SBPH revealed that rearrangements of genes in the mitochondrial genome could have occurred in the same species. On account of the different locations of trnH gene, the start and end codon of its neighboring genes had been changed. Even the same tRNA gene (e.g. trnS1 (AGN) and trnS2 (UCN) genes) could form different secondary structures in these two different individuals of SBPH. All these characteristics reveal that great divergence of mitochondrial genome could occur in different individuals of a species.The rearrangement of mitochondrial genome of planthoppers reported in this study adds more examples of the rearranged pattern of genes in the mitochondrial genomes of hemipteran insects studied so far. The fact that the rearrangement was conserved in Delphacidae suggests that gene rearrangements of mitochondrial genome probably occurred before the differentiation of this family. Compared with other hemipterans and other insects, the extreme short lengths of Laodelphax striatellus  and Nilaparvata lugens  for mitochondrial genome sequence were collected in China. Other natural populations which were used to validating location of trnH of SBPH and detecting the variable number of tandem repeats (VNTRs) in control regions of BPH and SBPH were collected in different locations of China  generated from the PCR-free random shotgun-sequencing libraries of average insert size 500 bp, one scaffold composed of three contigs of mitochondrial genome of SBPH  and one scaffold composed of four contigs of mitochondrial genome of BPH  were generated  were determined based on alignments and secondary structures of rRNA sequences of other hemipteran species. The boundaries of the ribosomal rrnL gene were assumed to be delimited by the ends of the trnL1 (CUN) and trnV. The 3′ end of rrnS gene was assumed to be delimited by the start of trnV while the 5′ end was determined through comparison with orthologous genes of other hemipteran species sequenced so far. The base composition, the Relative Synonymous Codon Usage (RSCU) values of BPH and SBPH were calculated with MEGA 4 program [After finished the gap closing work, we annotated the mitochondrial genomes of BPH and SBPH. All PCGs were identified by ORF Finder implemented at the NCBI website with the invertebrate mitochondrial genetic codes. Finally pair-wise comparisons with orthologous proteins from other hemipteran insects were performed to better define the limits of PCGs. The transfer RNA (tRNA) genes were identified using both of the tRNAscan-SE  and ARWE program .trnH rearrangement types really exist in natural populations, we sequenced a region of mitochondrial genome encompassing the trnH gene from 309 individuals of SBPH which were collected from 15 natural populations and also checked the Wolbachia infection status of each individual  to calculate the frequency of each haplotype of the sequence of this region. The sequences of each haplotype are deposited in the GenBank database under the Accession No. KC006945 - KC006960.After downloaded other mitochondrioal genome sequence  were not calculated for the variant length of these genes in these two individual mitochondria.The mutation rates of each gene between mitochondrial genomes of two individuals of Click here for fileNilaparvata lugens and Laodelphax striatellus used in this study. Collection information for natural populations of Click here for file


S)-PD172938 and arylated analogues with hypnotic sedative activity, obtained in good overall total yield (50%) and high enantiomeric purity (95% ee). The synthetic routes developed herein are particularly convenient in comparison with the current methods available in literature and are particularly promising for large scale applications.New bifunctional chiral ammonium salts were investigated in an asymmetric cascade synthesis of a key building block for a variety of biologically relevant isoindolinones. With this chiral compound in hand, the development of further transformations allowed for the synthesis of diverse derivatives of high pharmaceutical value, such as the Belliotti ( The enantiomeric excess was determined by derivatization of the compound into methyl ester 12 or amide 16.File 11H NMR, 13C NMR and HPLC traces.Complete experimental details and procedures, spectroscopic data, copies of


Azadirachta indica L.) leaf has been widely used in ayurvedic system of medicine for fertility regulation for a long time. The molecular mechanism by which neem leaf regulates female fertility remains poorly understood. Animal studies suggest that aqueous neem leaf extract (NLE) induces reactive oxygen species (ROS) - mediated granulosa cell apoptosis. Granulosa cell apoptosis deprives oocytes from nutrients, survival factors and cell cycle proteins required for the achievement of meiotic competency of follicular oocytes prior to ovulation. Under this situation, follicular oocyte becomes more susceptible towards apoptosis after ovulation. The increased level of hydrogen peroxide (H2O2) inside the follicular fluid results in the transfer of H2O2 from follicular fluid to the oocyte. The increased level of H2O2 induces p53 activation and over expression of Bax protein that modulates mitochondrial membrane potential and trigger cytochrome c release. The increased cytosolic cytochrome c level induces caspase-9 and caspase-3 activities that trigger destruction of structural and specific proteins leading to DNA fragmentation and thereby oocyte apoptosis. Based on these animal studies, we propose that NLE induces generation of ROS and mitochondria-mediated apoptosis both in granulosa cells as well as in follicular oocyte. The induction of apoptosis deteriorates oocyte quality and thereby limits reproductive outcome in mammals.Neem ( Azadirachta indica L.) has been considered as one of the most important medicinal plants worldwide. The medicinal utility of this plant are listed in ancient documents ‘Charak-Samhita’ and ‘Susruta-Samhita’ that are considered as the foundation of the Indian system of natural treatment, Ayurveda  during final stages of folliculogenesis and ovulation, while their effects are neutralized by active enzymatic antioxidant system  in rat ovary  of NLE  leaf extract-induced egg apoptosis in rat” funded by Department of Science and Technology, Ministry of Science and Technology, Government of India, New Delhi.Author SKC is the Principal Investigator of a project entitled “Studies on neem (


Myocardial infarction was experimentally induced in rat hearts and harvested immediately, 7, 14 and 28 days after the infarction induction. Anterior wall infarct samples underwent biaxial tensile and uniaxial compressive testing. Orientation of collagen fibres was analysed following mechanical testing. In this paper, we present the tensile and compressive stress–strain raw data, the calculated tensile and compressive moduli and the measured angles of collagen orientation. The presented data is associated with the research article titled “Characterisation of the mechanical properties of infarcted myocardium in the rat under biaxial tension and uniaxial compression”  [1]. Specifications TableValue of the data•Full presentation of biaxial mechanical data and collagen fibre orientation for healing myocardial infarcts.•The presented data demonstrates the mechanical and structural anisotropy of the healing myocardial infarcts.•First report to characterize the compressive properties of the healing rat myocardial infarcts.•Presentation of collagen fibre orientation in healing myocardial infarcts.•The presented mechanical and structural data can be utilised in constitutive modelling of healing myocardial infarcts for rats.1The data being shared describe the mechanical and structural properties of healing rat myocardial infarcts from individual samples in several post-infarct time points: immediately (i.e. 0 day), 7, 14 and 28 days after the induction of myocardial infarction. The mechanical data are composed of biaxial tensile stress–strain relationships , Fig. 4,2The details of the experimental work and materials are provided in


To the Editor: Marburg virus (MARV) and Ravn virus (RAVV), collectively called marburgviruses, cause Marburg hemorrhagic fever (MHF) in humans. In July 2007, 4 cases of MHF  occurred in miners at Kitaka Mine in southern Uganda. Later, MHF occurred in 2 tourists who visited Python Cave, ≈50 km from Kitaka Mine. One of the tourists was from the United States (December 2007) and 1 was from the Netherlands (July 2008); 1 case was fatal (Rousettus aegyptiacus bats (Egyptian fruit bats).Longitudinal investigations of the outbreaks at both locations were initiated by the Viral Special Pathogens Branch of the Centers for Disease Control and Prevention  in collaboration with the Uganda Wildlife Authority (UWA) and the Uganda Virus Research Institute (UVRI). During these studies, genetically diverse MARVs and RAVVs were isolated directly from bat tissues, and infection levels of the 2 viruses were found to increase in juvenile bats on a predictable bi-annual basis  Figure. R. aegyptiacus colonies in the area. Although minor colonies of small insectivorous bats were found, the only identifiable colony of R. aegyptiacus bats was found inside the re-opened Kitaka Mine, albeit at much reduced size, perhaps 1%–5% of that found before depopulation efforts.In October 2012, the most recent known marburgvirus outbreak was detected in Ibanda, a town in southwest Uganda. Ibanda is ≈20 km from the Kitaka Mine and is the urban center that serves smaller communities in the Kitaka area. This MHF outbreak was the largest in Ugandan history: 15 laboratory-confirmed cases occurred  were positive for marburgvirus RNA by quantitative reverse transcription PCR (32/233 [13.7%] adults and 21/167 [12.6%] juveniles; Of the 400 Phylogenetic analysis of viral RNA genome fragment sequences in this study showed high marburgvirus genetic diversity, including the presence of RAVVs and MARVs. Sequences for isolates from 3 bats were nearly identical to those of the MARV isolates obtained from patients in the 2012 Ibanda outbreak (Photographs taken during August 2008–September 2009 of bat extermination efforts at Kitaka Mine, and table showing demographic characteristics of bats captured during a Marburg hemorrhagic fever outbreak investigation at the mine in November 2012, Uganda.


Although extensively studied and indeed emotionally discussed for more than two decades the role of C-reactive protein (CRP) in cardiovascular disease remains controversial. Three major questions are still not yet resolved. (1) Is CRP a clinically relevant marker of cardiovascular risk? (2) Is CRP even more than a risk marker, that is, a risk factor in cardiovascular disease? (3) Finally, is CRP a cardiovascular drug target?This special issue comprises four review articles and three research articles that reflect the ongoing controversy over the subject. The review articles discuss CRP as a cardiovascular risk marker, CRP in animal models, CRP in its native and nonnative conformation, and CRP in human arteriosclerosis. The research articles deal with CRP as a cardiovascular risk marker in non-ST elevation acute coronary syndrome and with CRP as a drug target. The discussion climaxes in two research articles describing the use of CRP specific antisense oligonucleotides (ASOs) for the treatment of cardiovascular disease in animal models. The conclusions are indeed contradictory. When looking at the two articles, three general points may have to be taken into consideration. (1) Is the statistical power of the animal experiments adequate to draw definitive conclusions? (2) Are there off-target effects of the ASOs that may confound the results? And (3) is it necessary to design studies incorporating multiple on-target ASOs?Specific CRP inhibition followed by use of CRP inhibitors in controlled clinical trials may be the only way to prove or disprove a causative role for CRP in cardiovascular disease.


To the Editor: The earliest clinical cases of Zika virus infection were reported from continental South America in 2015  tests specific for dengue . Of these 50 patients, 30 (60%) were female. Most of these patients reported illness onset during December 9–27, 2015  were female. A total of 22 (8%) samples were positive for dengue; 2 were positive for chikungunya. Of the remaining 252 patients, 50 (20%) had 27, 2015  Figure 1The most commonly reported signs and symptoms were fever (86%), exanthema (72%), and headache (62%). The clinical characteristics of these infections showed no statistically significant difference with those associated with dengue and chikungunya virus infections and with cases found to be negative for all 3 viruses, suggesting that the negative cases could represent Zika virus infections  Table. OBy using Vero E6 cells (American Type Culture Collection), we isolated Zika virus from 9 samples . Phylogenetic analysis of 5 Zika virus sequences  placed these isolated (GenBank accession nos. KU724096–100) within the Asian lineage, the lineage involved in the spread of Zika virus in the Americas  Figure 2By using molecular methods, we confirmed diagnoses in 27% of patients during this outbreak. The distribution of positive results suggests that Zika virus was the predominant etiologic virus in this cohort, but we cannot firmly conclude this because most specimens tested negative for Zika, dengue, and chikungunya viruses.Although results from patient sampling and laboratory testing are not comparable, an assessment in Puerto Rico was able to detect Zika virus RNA by rRT-PCR or IgM by ELISA in 19% of 155 patients with suspected Zika virus infection (Several reasons might exist for the high proportion of specimens testing negative for Zika virus. Viremia is often low and short-lived in persons infected with Zika virus (The Panama Ministry of Health is following up with known pregnant women of the Guna Yala region who report Zika virus infection symptoms and is testing urine samples by using Zika virus–specific rRT-PCR within 14 days of symptom onset. Pregnant women confirmed to have Zika virus infection will receive ultrasound monitoring; however, the test has relatively low positive predictive value for detecting microcephaly (Distribution of symptoms by patients with Zika, dengue, and chikungunya virus infections in the Guna Yala region of Panama, November 27, 2015–January 22, 2016; epidemiologic curve for the outbreak of Zika virus infection in this region; and maximum-likelihood tree of the Zika virus isolated during the outbreak.


Alcohol screening and brief interventions (SBI) have a history and good evidence of efficacy in primary care settings . EfficacThis study aimed to discuss, and present for debate, challenges and opportunities relating to alcohol SBI in new settings from published and previously unpublished studies of recent SBI implementation in Scotland and England.A narrative review was conducted of evidence from diverse studies including research into training and implementation of alcohol SBI outside of primary care .The challenges and opportunities can be conceptualised in terms of questions that should be asked before considering implementation of alcohol brief interventions in a new setting: (1) Is there a need for SBI delivery in this service? (2) How are practitioners currently addressing alcohol use - how when, where does SBI fit in? (3) Will SBI work/do harm in this setting? (4) Will SBI be perceived as legitimate by practitioners and acceptable to service-users? (5) Will practitioners have the ability to deliver SBI? (6) What support will be needed? (7) What will support routine implementation of SBI in this setting?Careful consideration should be given to every aspect of the design, purpose, context and evaluation of alcohol SBI in any given setting preferably by robust stepwise research prior to widespread implementation. Avoiding assumptions, including about screening methods and intervention goals, is likely to be important for effectiveness, implementation and avoiding unintended harms.


The body sodium imbalance (dysnatremias) may be associated with increased mortality of critically ill patients. Evidence suggests that changes in the serum sodium level on admission to the intensive care unit (ICU), may lead to a poor outcome.The objective of this study was to evaluate the ability of APACHE II and serum sodium levels to predict mortality of surgical critically ill patients.One hundred and ninety-five surgical patients  admitted to the ICU in the post-operative phase were retrospectively studied. The patients were divided into survivors (n=152) and nonsurvivors (n=43). APACHE II, and serum sodium levels at admission, 48-h and discharge were recorded. The capability of each index  to predict mortality of surgical patients was analysed by receiver-operator characteristic curves (ROC). Odds ratios (OR) and 95% confidence interval (CI) for hyponatremia (Na< 136 mmol/L) and hypernatremia (Na>144 mmol/L) were calculated.Comparison of data between survivors and nonsurvivors is summarized in Table Even though APACHE II was the most effective index to predict mortality in the surgical critically ill patients, the serum sodium levels on admission may also be used as predictor of outcome.


Chronic migraine (CM) affects 2% of the population and Botox is the only licensed treatment for prevention of adult patients with CM.In the UK, National Institute for Clinical Excellence (NICE) approved its use on the NHS in patients who failed three preventive medications.NICE recommends continuing treatment beyond cycle 2 in those with 30% reduction in headache days (negative stopping rule) and the treatment is stopped when migraine become episodic (positive stopping rule).However, the long-term outcome ie duration of required treatment remains uncertain.To ascertain the duration of treatment with Botox for the prevention of chronic migraine in responders as per NICE criteria.Adult patients with CM attending the Hull migraine clinic were offered Botox based on clinical needs and maintained a headache diary.Data were extracted for headache, migraine, and headache-free daysResponder rate was assessed applying NICE criteriaPatients (N=67) who commenced treatment between July 2010 and June 2012 were followed up for at least 18 months.30 Patients stopped treatment at Cycle 2 as per negative stopping rule by NICEOf 37 responders, only 12 were still on treatment at 18 months. Nearly half of the patients stopped treatment by the 4th Cycle (positive stopping rule).We continue to follow all patients who receive Botox for prevention of chronic migraine and aim to present 2 year follow up on a larger cohort at the EHMTIC September 2014.


Electronic medical records (EMR) form a rich repository of information that could benefit public health. We asked how structured and free-text narrative EMR data should be combined to improve epidemic surveillance for acute respiratory infections (ARI).Eight previously characterized ARI case detection algorithms (CDA) were applied to historical EMR entries to create authentic time series of daily ARI case counts (background). An epidemic model simulated influenza cases (injection). From the time of the injection, cluster-detection statistics were applied daily on paired background+injection (combined) and background-only time series. This cycle was then repeated with the injection shifted to each week of the evaluation year. We computed: a) the time from injection to the first statistical alarm uniquely found in the combined dataset (Detection Delay); b) how often alarms originated in the background-only dataset ; and c) the number of cases found within these false alarms (Caseload). For each CDA, we plotted the Detection Delay as a function of FAR or Caseload, over a broad range of alarm thresholds.CDAs that combined text analyses seeking ARI symptoms in clinical notes with provider-assigned diagnostic codes in order to maximize the precision rather than the sensitivity of case-detection lowered Detection Delay at any given FAR or Caseload.An empiric approach can guide the integration of EMR data into case-detection methods that improve both the timeliness and efficiency of epidemic detection. Epidemics of acute respiratory infections (ARI), whether due to influenza Electronic data offer the opportunity for more timely and complete gathering of health information compared to what has historically been achieved through manual, paper-based reporting To gain insight on the conduct of surveillance in an EMR environment, we previously evaluated how EMR entries should be assembled to discover individuals with ARI The Institutional Review Boards of the Veterans Administration (VA) Maryland Health Care System and the University of Maryland approved this study. The study was granted a waiver of consent as risks were limited to information confidentiality and the work would not have otherwise been feasible, given the large number of EMR records screened for possible ARI. All EMR information was anonymized and de-identified prior to simulations and analyses, which used only daily case counts.Historical EMR data were extracted from the Veterans Integrated Service Technology Architecture (VistA) repository using the MDE software  and transferred to a Structured Query Language (SQL) relational database .th Revision, Clinical Modification, ICD-9) either used by the original Centers for Disease Control and Prevention (CDC) “BioSense” surveillance system Outpatients with possible ARI were identified by applying previously developed ARI case-detection algorithms (CDAs) et al. To create a plausible ARI outbreak to be discovered by the surveillance system, we developed an epidemic model of influenza . The model included 30 contiguous ZIP codes centered on Baltimore, Maryland, and consisted of a coupled series of differential equations to describe the overall epidemic http://www.r-project.org). Starting on the day when the synthetic epidemic was injected into a CDA-specific authentic background time series, and then daily for a total of 80 days, a statistical outbreak detection method (see below) was applied in parallel to corresponding time series that included either: a) both background and epidemic cases  W2c We computed three performance benchmarks at any given statistical alert threshold: 1) the Detection Delay, the time from the injection of the synthetic outbreak to the first true positive alarm, averaged for the 52 surveillance cycles of the evaluation year; 2) the false alarm rate (FAR), the average daily number of unique alarms issued in the background-only time series during the evaluation year; 3) the Caseload, defined as the total yearly number of cases included in the above false alarms. Corresponding Detection Delays, FARs and Caseloads were obtained for a range of statistical alert thresholds adjusted iteratively to focus on a FAR range felt to be of practical use for surveillance i.e. 0–10%.The activity monitoring operating characteristic (AMOC) curves The surveillance advantages of adjusting the ICD-9 codes used to identify ARI cases can be visualized by comparing the AMOC curves obtained using a “respiratory” code set used by a surveillance system of national scope Case detection that relied solely on text analyses of clinical notes resulted in outbreak detection performance roughly on par with that of the VA-adapted set of ICD-9 codes  Time series of background casecounts. 8-year background time series are provided for each of the CDA outlined in the (TXT)Click here for additional data file.Datafile S2(CDA 2) Time series of background casecounts.(TXT)Click here for additional data file.Datafile S3(CDA 3) Time series of background casecounts.(TXT)Click here for additional data file.Datafile S4(CDA 4) Time series of background casecounts.(TXT)Click here for additional data file.Datafile S5(CDA 5) Time series of background casecounts.(TXT)Click here for additional data file.Datafile S6(CDA 6) Time series of background casecounts.(TXT)Click here for additional data file.Datafile S7(CDA 7) Time series of background casecounts.(TXT)Click here for additional data file.Datafile S8(CDA 8) Time series of background casecounts.(TXT)Click here for additional data file.


A subset of patients with autoimmune diseases including rheumatoid arthritis (RA) and lupus appear to be exposed continually to interferon (IFN) as evidenced by elevated expression of IFN induced genes in blood cells. In lupus, detection of endogenous chromatin complexes by the innate sensing machinery is the suspected driver for the IFN, but the actual mechanisms remain unknown in all of these diseases. We investigated in two randomized clinical trials the effects on RA patients of baminercept, a lymphotoxin-beta receptor-immunoglobulin fusion protein that blocks the lymphotoxin-αβ/LIGHT axis. Administration of baminercept led to a reduced RNA IFN signature in the blood of patients with elevated baseline signatures. Both RA and SLE patients with a high IFN signature were lymphopenic and lymphocyte counts increased following baminercept treatment of RA patients. These data demonstrate a coupling between the lymphotoxin-LIGHT system and IFN production in rheumatoid arthritis. IFN induced retention of lymphocytes within lymphoid tissues is a likely component of the lymphopenia observed in many autoimmune diseases.NCT00664716.ClinicalTrials.gov  Systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), Sjogren’s syndrome, systemic sclerosis, myositis and multiple sclerosis patients have circulating blood cells with elevated levels of RNA from IFN-induced genes, i.e. an ‘IFN signature’ We have investigated the effects of inhibition of the lymphotoxin-LIGHT system in RA using a soluble lymphotoxin-beta receptor  immunoglobulin fusion protein called baminercept. LTBR is a central component of a signaling system whereby lymphocytes instruct stromal cells to differentiate into specialized vasculature and certain reticular networks Unexpectedly, we found that baminercept reduced the IFN signature in RA patients. The reduced IFN signature in RA patients following baminercept treatment is the first time outside of high dose steroid therapy that an IFN signature was decreased by a pharmacological treatment not targeting IFN itself. Taken together with the known effects of LTBR inhibition, these studies not only link the LTBR axis to IFN production in man, but also provide potential insight into the nature of the IFN signature.Two randomized phase IIb controlled studies of the effects of baminercept in rheumatoid arthritis were conducted. One study enrolled patients with an inadequate response to disease-modifying antirheumatic drug therapy (DMARD-IR) and the other involved patients with an inadequate response to tumor necrosis factor inhibition (TNF-IR)  and an IFN score calculated  and type II IFN (IFNγ) and, furthermore, each IFN is capable in many contexts of inducing the expression of the other IFN class Elevated levels of type I IFN or an IFN-like inducing activity can be found in the sera of a subset of the SLE patients with a transcriptional IFN signature; however, in RA the results range from not detectable to low levels relative to SLE sera SLE patients with a high IFN signature tend to be lymphopenic In rodents, blockade of the LTBR system leads to lymphocytosis within several weeks most likely due to loss of high endothelial venule addressin expression and reduced entry into the lymph nodes and mucosal environments In mice, lymphocytosis following LTBR blockade results from elevated T and B cells counts Given the emphasis on plasmacytoid DC as the source of the IFN signature in SLE, we examined RNA levels in the blood of two known pDC markers, CLEC4C (BDCA2) and LILRA4 (ILT7). LILRA4 levels increased following baminercept treatment, yet CLEC4C levels were unchanged  scoring system. Overall, baminercept was well tolerated  was a multicenter, phase IIb, double-blinded, placebo-controlled study of RA patients who lacked an adequate response to TNF-blocking therapy and had discontinued TNF blocking treatment for at least 90 days. The study dosed 114 patients with subcutaneous injections q2w of either placebo (38) or baminercept 200 mg (76). This study was terminated early due to poor efficacy in the DMARD-IR study; however, 81 patients completed the 3 months of dosing and another 15 patients received at least 2 months of treatment. Study was conducted between March 2007 and October 2008 at 40 sites in the United States, Canada, Belgium and United Kingdom.Investigators for both baminercept studies are listed in the supplemental materials in SLE data were from registry  representing a collection of 292 SLE patients from the Hopkins Lupus Center in the US. Patients were eligible if they were aged 18–75 years and met the American College of Rheumatology Revised Criteria for Classification of Systemic Lupus Erythematous. Baseline data were used in this study and patients were under standard clinical practice. Normal controls for RNA analyses were composed of healthy volunteer donors from Biogen Idec. Control group had equal numbers of males and females and was not exactly gender balanced with the predominantly female composition of both the RA and SLE cohorts.The RA studies were approved by the appropriate institutional review boards or ethics committees and all patients provided written informed consent Click here for additional data file.Checklist S1CONSORT Checklist.(PDF)Click here for additional data file.Protocol S1Trial Protocol.(PDF)Click here for additional data file.Protocol S2Trial Protocol.(PDF)Click here for additional data file.


Preoperative quantitative assessment of coronary angiography (CAG) and real-time graft size and graft flow assessment using epigraftic ultrasonography in the operative field have been performed to ascertain the patency in the right coronary artery bypass grafting (CABG).This study aims to evaluate the efficacy of the quantitative coronary artery and graft assessment to increase the patency of the right CABG.CABG was performed in 200 patients from January 2010 to December 2014. Ninety-three patients underwent all of preoperative CAG, real-time graft assessment and postoperative CAG with a total of 93 grafts anastomosed to the right coronary artery. Severity of coronary artery stenosis was evaluated by CAG. The size of target right coronary artery was measured by ultrasonography in the operative field. Parameters about graft flow were obtained from flow velocity curve in all the graft. The grafts were divided into two groups: patent grafts  and failing grafts . All factors were compared in these two groups and evaluated by logistic analysis and receiver operating characteristic (ROC) curve analysis.The overall patency as measured by postoperative CAG was 94.6% (88/93). There were 7 failing grafts including 5 occlusion and 2 competitive slow flow. Logistic regression analysis revealed that the percentage of graft size measured by ultrasonography divided by the size of the right coronary artery (graft-RCA size mismatch) and the pulsatility index (PI) were independent predictors of early graft failure . ROC curve analysis revealed that graft-RCA size mismatch > 2.08 and PI > 4.55 were predictors of graft failure.In this series, epigraftic ultrasonography depicted graft flow clearly. Combination of preoperative quantitative coronary artery assessment and real time graft assessment was essential to predict graft failure during the CABG. This technique may increase the patency of the right coronary artery bypass grafting.


The condition accounts for almost half of all nail issues yet continues to be largely under-reported and untreated. This study will investigate the antifungal efficacy of eucalyptus oil A longitudinal prospective study was used to monitor the changes in toenail mycotic infections over a four-month period, using undiluted eucalyptus oil as a topical antifungal agent. Data were collected from 22 participants (14 men and 8 women) aged between 40 and 84 with a total of 70 toenails, 35 with nail matrix infection and 35 without. To review the effect of eucalyptus oil on the clinical appearance of the fungal infected nail plates, toenails were monitored at four-weekly intervals for a period of four months. The participants’ satisfaction with the therapy was assessed using the OnyCOE-t questionnaire.Of the 70 nails analysed, the patterns of infection were classified as proximal subungual onychomycosis (49%), distal lateral subungual onychomycosis (47%), white superficial onychomycosis (3%), and total dystrophic (1%). Almost half of all participants (45%) had onychomycosis affecting only one toenail. Only 23% of participants had previously treated the infection and the mean duration of infection was 10 years and 3 months.Nails with superficial onychomycosis (n=2) were found to have 86% clearance of infection after four months. One third of all nails (n=11) with distal lateral subungual onychomycosis and 50% (n=17) of nails with proximal subungual onychomycosis demonstrated a zone of clearance at the proximal nail plate tissue, suggesting a fungistatic effect of the eucalyptus oil.Topical eucalyptus oil is more effective as an antifungal treatment for fungal infected toenails without nail matrix infection. Eucalyptus oil may provide an acceptable and cheaper alternative to prescription topical antifungal agents, for people with white superficial onychomycosis or distal lateral subungual fungal nail infections.


UGT1A1*28 was not questioned in our study.There is an error in the second sentence in the third paragraph of the “Genetic differentiation and haplotype diversity for DMET Plus markers” section of the Results. The correct sentence is: Although, the array captures many of the functional variants described for these genes, unfortunately The S5 TableMinor Allele Frequencies (MAF) for 1,647 genetic markers. The markers are those included in the DMET Plus array and for all populations: Europeans (Hapmap CEU), Africans (Hapmap YRI and LWK), Native Americans (Zapotecas), Brazilians and Mexican mestizos (Guanajuato (GUA); Guerrero (GUE); Sonora (SON); Veracruz (VER) and Yucatan (YUC)).(XLSX)Click here for additional data file.


Systematic review and meta-analysis of published studies of alcohol use among young people (age 15–24 years) in eastern Africa to estimate prevalence of alcohol use and determine the extent of use of standardised screening questionnaires in alcohol studies.2 statistic (DerSimonian-Laird).Five databases  were searched for publications until 30th June 2013. Results were summarised using the guidelines on preferred reporting items for systematic reviews and meta-analyses (PRISMA) and on quality assessment using the modified quality assessment tool for systematic reviews of observational studies (QATSO). Heterogeneity was assessed using the IWe identified 2785 potentially relevant studies, of which 56 were eligible for inclusion. Only two studies (4%) used the standardised Alcohol Use Disorder Identification Test (AUDIT) questionnaire, and six studies (13%) used the Cut down, Annoyed, Guilt, Eye opener (CAGE) questionnaire. The reported median prevalence of alcohol use was ever-use 52% [interquartile range (IQR): 20–58%], use in the last month 28% (IQR: 17–37%), use in the last year 26% (IQR: 22–32%), and problem drinking as defined by CAGE or AUDIT 15% (IQR: 3–36%). We observed high heterogeneity between studies, with the highest prevalence of ever use of alcohol among university students  and female sex workers . Current use was most prevalent among male sex workers .Reported alcohol use and problem drinking were common among diverse groups of young people in eastern Africa, indicating the urgent need for alcohol-focused interventions in this population. Few studies have used standardised alcohol screening questionnaires. Epidemiological research to investigate alcohol-focused interventions in young people should aim to apply such questionnaires that should be validated for use in this population. Harmful alcohol use is a significant public health problem that often begins early in adult life. Globally, an estimated 2 billion people drink alcohol and 76 million have alcohol use disorders (AUD) WHO . The meaet al. et al. et al. et al. et al. et al. Factors associated with alcohol use include religion, personal income, education level, peer influence, having older sexual partners, stress and relatives and friends using alcohol  estimate the prevalence of alcohol use among specific groups of young people (15–24 years) in eastern Africa; (ii) determine the extent of use of standardised alcohol screening questionnaires [Alcohol Use Disorder Identification Test (AUDIT), Cut down, Annoyed, Guilt, Eye opener (CAGE) in identifying alcohol use and AUD in this region; (iii) assess the quality of research papers included in the review; and (iv) describe factors associated with initiation and persistence of alcohol use among young people in eastern Africa.).Five databases  were searched for publications to 30th June 2013. We used the following key terms:  AND (young people OR adolescent OR teenage OR youth) AND . , and consensus on potential eligibility reached. Studies were eligible if they were conducted in eastern Africa ; and included prevalence of alcohol use for young people aged 15–24 years.et al. et al. Guidelines on preferred reporting items for systematic reviews and meta-analyses (PRISMA) were used (Moher We used a data extraction form to collect the following information from each eligible article: (i) country; (ii) year the study was conducted; (iii) year of publication; (iv) study population ; (v) sample size; (vi) definition of alcohol use ; (vii) prevalence of alcohol use and AUD (problem drinking as classified by CAGE and AUDIT); (viii) factors associated with the initiation and persistence of alcohol use; (ix) alcohol use screening questionnaires applied; and (x) complications associated with alcohol use.et al. A descriptive quality assessment of the final papers included in the meta-analysis was conducted using the modified quality assessment tool for systematic reviews of observational studies (QATSO) (Wong 2 statistic (DerSimonian-Laird) and reported the prevalence for studies in four groups: (i) ever use of alcohol; (ii) alcohol use in the last year, (iii) alcohol use in the last month (current use) and (iv) problem drinking as defined by CAGE and AUDIT .We identified 4013 published study citations from five databases, of which 1228 were duplicates. Thus, 2785 abstracts were screened for initial eligibility to identify studies conducted in eastern Africa. We identified 696 relevant abstracts of studies conducted in eastern Africa. We conducted further screening for studies reporting on alcohol use and identified 285 abstracts for full article review. Of these 285 abstracts for full article assessment, we could not access six full articles, 11 were review articles and five were conference posters.Thus, we reviewed 263 full-text papers and identified 56 eligible for inclusion in the review. The main reason for exclusion was that the paper did not report information on alcohol use from the target population, that is, young people aged 15–24 years, or that young people were included but we could not separate the prevalence in this age group from that in older people , and two were case–control studies. Almost all studies  were conducted in four countries: Ethiopia (n = 19), Kenya (n = 15), Tanzania (n = 10) and Uganda (n = 8); the remaining four studies were conducted in Rwanda (n = 2), Seychelles (n = 1) and Eritrea (n = 1). Most studies  reported current alcohol use, 17 studies reported ever use of alcohol, four studies reported alcohol use in the last year, and five studies reported problem drinking. Only two studies used the AUDIT alcohol use screening questionnaire  used probability-based sampling and had a response rate above 80% . However, 17 studies did not report the response rate  and street children (21% vs 11%) and high in male university students (53% vs 50%). There was significant heterogeneity based on I2 statistics in all subgroups, and therefore, we do not report pooled prevalence.Figure vs 17%).Four studies reported the prevalence of alcohol use in the last 12 months. These showed similar levels as found for reported current alcohol use. Three studies were from the general population  with a pooled prevalence of 30% (95%CI: 27–33%) and one from university students . One study reported gender-specific prevalence; it was high among males (34% vs. 9%(IQR: 9–20%) in the general population, 28% (IQR: 13–44%) vs 19%(IQR: 7–32%) among healthcare service attenders, 60% (IQR: 56–63%) vs. 41% (IQR: 24–58%) in secondary schools, and 43% vs. 28% in a university. There was no significant heterogeneity in studies conducted among university students except one study among female university students  among female bar workers and 47% (95%CI: 40–53%) among male sex workers. Median prevalence in the general population was 3% (IQR: 1–15%); two of the studies in the general population were from Ethiopia and showed low levels of problem drinking (1–3%), but a study from Tanzania reported a rather high median prevalence of 15% (95%CI: 10–20%).et al. et al. et al. In this review, eight studies reported problematic drinking, two studies applied AUDIT, and six (13%) used CAGE to screen for problem drinking; two of the studies that applied CAGE and one study that applied AUDIT did not report scores according to age groups  and female sex workers (66%) and lower among the general population and primary school students. Few studies reported alcohol use in the last year, and median prevalence in the general population was 29%. Reported current alcohol use was highest among male sex workers (69%), followed by the university students (33%), and was lowest in the general population and secondary school students with the exception of one study in the Seychelles that reported a high prevalence of 61%. Problem drinking was highest among groups known to engage in high-risk behaviours (such as bar workers and sex workers). Generally, reported alcohol use across all definitions of use was highest among groups known to engage in high-risk behaviours, followed by university students. Individuals attending healthcare services and general populations and secondary school students reported the lowest prevalence. Studies included in this review were of good quality; however, about two-thirds employed face-to-face interviewing approaches, an approach prone to social desirability bias that could lead to underreporting of alcohol use.vs. students vs. sex workers). We also attribute these variations to social influence and peer pressure ; it is, however, not widely used for the assessment of AUD among young people in general populations. In our review, only two studies used the internationally recommended AUDIT alcohol screening questionnaire (Mbatia et al. et al. The use of effective and validated instruments for the screening and assessment of alcohol use is essential to guide research and is important for the design and evaluation of interventions. AUDIT is validated and recommended by WHO for use at the primary healthcare settings and for the assessment of AUD in developing countries (Saunders Reported alcohol use among young people in eastern Africa is common and varies between different populations. The prevalence of AUD was highest among populations known to engage in high-risk sexual behaviours, but was also high among students in some of the studies. The studies reviewed lacked data about initiation and persistence of alcohol use, and little information was available about risk factors associated with alcohol use, and AUD. Notably, only few of the studies reviewed used internationally recommended and validated screening questionnaires such as AUDIT. Future epidemiological studies on alcohol use among young people should apply these questionnaires to facilitate comparison. However, such questionnaires have not been evaluated among young people in Africa, and studies closing this knowledge gap are therefore also required. Future studies should also determine factors responsible for initiation, persistence, and patterns of use in preparation for potential interventions. There is an urgent need of targeted interventions for groups of young people with a particularly high risk of alcohol use and AUD such as college students and young sex workers.


Background Exposure to high levels of Aluminium (Al) leads to a neurodegenerative disorders , which may be mediated through over generation of free radicals. So in the present study we investigated the ability of both Quercetin and Omega 3 to ameliorate Al adverse effect on brain antioxidant through monitoring the main brain antioxidant enzymes on molecular and cellular levels.Forty male albino rats were used, they were divided into 4 groups; Control, Aluminum Chloride (AlCl3) supplemented group that orally supplemented with 100mg of AlCl3  per Kg b.w. for two months. Quercetin group treated as ALCl3 group and orally supplemented with 100 mg/kg b.w. Quercetin for two months according to Hui et al. [Our results indicate a significant increase in superoxide dismutase (SOD) activity and MDA level and a significant decrease in the activities of catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPX) and levels of reduced glutathione (GSH) in brain tissues in AlCl3 supplemented group when compared with control or Quercetin and Omega 3 supplemented groups Table . At the Both Quercetin and Omega 3 has the ability to overcome the Al induced oxidative stress in brain, manifested by the significant reduction in free radicals concentration and induction of the activity and gene expression of the brain antioxidant enzymes.


Patients with inflammatory bowel disease (IBD) demonstrate an inflammatory response which bears some similarities to that seen in ischaemic heart disease (IHD). The nature of the association of IBD with IHD is uncertain. We aimed to define the extent and direction of that association.This retrospective cohort study examined records from patients aged ≥ 15 years with IBD from 1987–2009 (n = 19163) who were age and gender matched with patients without IBD (n = 75735) using the General Practice Research Database. The primary outcome was the hazard ratio for IHD.A higher proportion of IBD patients had a recorded diagnosis of IHD ever, 2220 (11.6%) compared with 6504 (8.6%) of controls. However, the majority  developed IHD prior to IBD diagnosis (1404 (63.2%) of IBD cases and 3090 (47.5%) of controls). There was increased IHD incidence in the first year after IBD diagnosis. Mean age at IHD diagnosis was statistically similar across all IBD groups apart from for those with Ulcerative Colitis (UC) who were slightly younger at diagnosis of angina compared to controls  and coronary heart disease . Of those developing IHD following IBD diagnosis, UC patients were at higher risk of IHD (unadjusted HR 1.3 (95% CI 1.1–1.5), p<0.001) or MI (unadjusted HR 1.4 (95% CI 1.1–1.6), p = 0.004).Although IHD prevalence was higher in IBD patients, most IHD diagnoses predated the diagnosis of IBD. This implies a more complex relationship than previously proposed between the inflammatory responses associated with IHD and IBD, and alternative models should be considered. The inflammatory bowel diseases (IBD) ulcerative colitis (UC) and Crohn’s Disease (CD) are disorders of uncertain origin. In both UC and CD, an exaggerated immune response to unknown antigens is evident  and a nuThere is a well-described association between rheumatoid arthritis (RA) and increased cardiovascular mortality, mainly as a result of ischaemic heart disease (IHD)., 10 AtheThere are sufficient shared characteristics to the inflammatory responses seen in IBD and RA to hypothesise that the recognized association between RA and IHD might also exist for IBD and IHD. Previous cohort studies have examined the risk of IHD in patients with IBD and have variously found no effect, or a slihttp://www.cprd.com/home/) between 1987 and 2009. For each subject with IBD, a further 4 subjects were identified and matched for age and sex but without IBD. All cohort members were followed from the start date (i.e. the date their GPRD record began) until the earliest occurrence of one of the following endpoints: recorded code of IHD, death, date of exit from GPRD registered GP practice, or end of study period (31.12.2009). Patients with less than (1) 2 years up-to-standard enrolment with the GP or (2) 1 year of computerized prescription history or (3) no health contacts during their complete period of follow-up were excluded from the cohort. Patients whose date of death or date of transfer out of a GPRD-registered practice preceded date of IBD diagnosis were excluded from the cohort. Patients <15 years of age at entry were excluded.The study population for this retrospective cohort study consisted of around 2.1 million male and female patients with an active registration at practices contributing to the General Practice Research Database , or Crohn’s Disease (CD), or inflammatory bowel disease (not otherwise specified) (IBD (nos)) recorded on the database since 1987; patients with ICD-10 Read Codes (K50-K51) records of both UC and IBD were coded as UC; CD plus IBD were coded as CD; individual records which included both UC and CD were included only in analyses of IBD (nos). Prevalent IBD cases on 01.01.1987 were excluded. The index date was defined as the date of first recording of IBD. Lifetime IBD severity was determined using an algorithm based on IBD drug-use  and surgical intervention (colectomy/stomata) developed by team members with clinical and research expertise in IBD.Age, sex and practice matched controls were allocated the same index date as their matched case. Because IHD has not been subjected to the same specific validation studies as IBD, our operational definition was developed using advice from clinicians with special interest in cardiovascular disease to include the following: Read Codes for IHD  Coronary Heart Disease (CHD) was considered for the purpose of this study to be synonymous with IHD and Read Codes for coronary heart disease (including coronary artery disease) were included.Data was accessed within limits set out by the Medical Research Council licence agreement for academic access with Medical Research Ethics Committee ethical approval. The proposal was approved by the Independent Scientific Advisory Committee of the GPRD (protocol number 10_003).a priori hypothesis, that IHD followed IBD, was conducted on a subset which included cases of IHD following IBD and their associated matched controls (COHORT B: n = 77540).Descriptive analysis of baseline characteristics and exploration of the direction of association between IBD and IHD was conducted using the whole cohort of patients aged over 15 years . Subsequent analysis of the The relative risks of ischaemic heart disease (IHD) and its sub-entities myocardial infarction (MI) and angina), as well as CHD were estimated as hazard ratios (HRs) for patients with IBD compared to non-IBD patients, using Cox proportional hazard models. Models were subsequently adjusted for age, gender, socio-economic status , comorbidity and concomitant drug use, IBD disease severity, and smoking status. We carried out both forward and backward stepwise covariate selection within the cox proportional hazard model. All covariates were initially included with entry testing based on the significance of the score statistic, and removal testing based on the probability of a likelihood-ratio statistic based on conditional parameter estimates. Models were tested for interactions between fitted variables. Precision of estimates is reported using 95% confidence intervals. Data management was performed using Stata 10IC. Cox regression proportional hazard computations and Kaplan-Meier survival analysis were conducted using SPSS 21.Data were examined from electronic primary care records from 221 general practices across the UK from patients with a first diagnosis of IBD recorded from 1st January 1987 to 31st December 2009 . FollowiOf 19163 IBD cases, a total of 2220 (11.6%) had an IHD diagnosis ever, compared with 6504 (8.6%) of those without IBD. Of these IHD diagnoses, 4494 (51.5%) occurred prior to IBD diagnosis (1404 (63.2%) of IBD cases and 3090 (47.5%) of controls) . AccordiA total of 17358 patients were excluded from subgroup analysis where IHD preceded IBD or, for controls, the matched IBD date  . Subset In order to examine the subset in a way that was consistent with previous research, the inciThis is the first study to fully examine both the strength and direction of association between development of ischaemic heart disease (IHD) and IBD. An increased incidence of IHD, angina, MI, and CHD diagnoses was seen in the first year after IBD diagnosis but the mean age of diagnosis comparing IBD cases with non IBD cases was not statistically different, apart from those with Ulcerative Colitis (UC) who were slightly younger at diagnosis of angina and coronary heart disease. Of those developing IHD following an IBD diagnosis, patients with UC were at slightly higher risk of a diagnosis of IHD or MI although results were not significant after adjustment for the role of covariates. Most notably, we found that although overall prevalence of IHD was higher in those with IBD, in the majority of cases the diagnosis of IHD actually predated that of IBD.This is the largest British cohort study to examine this issue. It used the General Practice Research Database which is an appropriate and reliable resource for IBD research, having been used for at least 19 studies published in peer-reviewed journals since 2000. The diagnostic reliability of IBD diagnosis and the year of IBD diagnosis in the GPRD has been confirmed in a recent validation study. Other GPAs others have highlighted , there iThe temporal relationship between IBD and IHD may be weakened by differences with which the diagnosis of each is recorded in the GPRD. IHD is a diagnosis made in the majority of cases fairly soon after the onset of symptoms, because of an event requiring admission to hospital . Nevertheless, among those with stable angina, nearly half have symptoms for 6 months or more before diagnosis. For IBD It is possible that IHD cases may have been missed or incorrectly coded within GPRD. Unrecorded diagnoses are increasingly unlikely within UK general practice due to policy and fiscal incentives to assess, monitor and treat cardiovascular disease, althoughThis is the first known study to examine the temporal relationship between IHD and IBD and we found that the majority of IHD cases predated IBD diagnosis. Previous research has predominantly focused attention on incident cases of IHD occurring at or after diagnosis of IBD. It is notable that the overall risk of prevalent (i.e. whole cohort) IHD was almost identical to a recent Danish study  at 2.1 IBy identifying and quantifying both the strength and direction of association between IBD and IHD, this study provides important insights into the burden of IHD in the IBD population. This is relevant for basic scientists exploring the temporal causal pathway, and clinicians who should be aware of the risks of IHD in patients with IBD and vice versa.S1 FigTotal number of patients showing time from IBD diagnosis (T0) to the first cardiovascular event  according to IBD type and age categories.(TIF)Click here for additional data file.S2 FigTotal number of patients showing time from IBD diagnosis (T0) to the first cardiovascular event  according to IBD type and gender categories.(TIF)Click here for additional data file.S3 FigTotal number of patients showing time from IBD diagnosis (T0) to the first Coronary Heart Disease (sub-group of CVD) event according to IBD type.(TIF)Click here for additional data file.S4 FigTotal number of patients showing time from IBD diagnosis (T0) to the first Ischaemic Heart Disease (sub-group of CVD) event according to IBD type.(TIF)Click here for additional data file.S5 FigTotal number of patients showing time from IBD diagnosis (T0) to the first Angina (sub-group of CVD) event according to IBD type.(TIF)Click here for additional data file.S6 FigTotal number of patients showing time from IBD diagnosis (T0) to the first Myocardial Infarction (sub-group of CVD) event according to IBD type.(TIF)Click here for additional data file.S7 FigTotal number of patients showing age at onset of Cardiovascular Disease  according to IBD type.(TIF)Click here for additional data file.


Our data provides first evidence that the genetic factors associated with pigmentation traits are risk loci of UM susceptibility.While the role of genetic risk factors in the etiology of uveal melanoma (UM) has been strongly suggested, the genetic susceptibility to UM is currently vastly unexplored. Due to shared epidemiological risk factors between cutaneous melanoma (CM) and UM, in this study we have selected 28 SNPs identified as risk variants in previous genome-wide association studies on CM or CM-related host phenotypes (such as pigmentation and eye color) and tested them for association with UM risk. By logistic regression analysis of 272 UM cases and 1782 controls using an additive model, we identified five variants significantly associated with UM risk, all passing adjustment for multiple testing. The three most significantly associated variants rs12913832 , rs1129038  and rs916977  are correlated (r BAP1, CDKN2A, or BRCA2 explain only about 3% of UM population-specific risk678310Uveal melanoma (UM) is the most common primary adult intraocular cancer142 > 0.5), which were still significant following adjustment for multiple testing  established in recent GWASs for association with CM or related host phenotypes (including skin and eye pigmentation) , in a po testing : rs12913esting 12. As showOCA2, which codes for P protein involved in melanin synthesisHERC2, is a key pigmentation “regulator allele” that impacts the expression of OCA2 via a long range enhancer mechanismOCA2 expression resulting in darkly pigmented melanocytes. The C-allele conversely reduces expression of OCA2 producing lightly colored melanocytes3The locus at 15q13.1 determines pigmentation of eyes and skin by regulating the expression of 2 = 0.48)  , that waIt is important to note that our study has limited power and hence may reduce the ability to detect associations exerting smaller risk effects. It is possible that there are other associations among the selected variants that are likely missed in this study due to relatively small size of our UM case population. The most significant associations at HERC2/OCA2 locus , as wellGNAQ mutation signatures were observed in posterior (choroid) versus anterior UM tumors , likely reflecting divergent UM pathways related to toxic pheomelanin synthesisAlso, the relatively small size of our case population did not allow for stratifying the associations by the primary location of tumors, and this will need to be explored in subsequent large consortia analyses. The molecular effect of pigmentation on different UM subtypes has recently been suggested, as different A limitation in this study is an inability to assess the effect of pigmentation phenotype information in our sample populations as the HERC2/OCA2 locus exerts a purifying selection effect in European ancestries21The identification of novel germline genetic loci involved in UM susceptibility in our study provides the first evidence of a link between the inherited genetics of pigmentation and UM risk. It has been established that lighter pigmentation and chronic sun exposure impact the development of choroid nevi, which occur in ~7% of the US population and are a known precursor for UMFor cases, the study originally employed 286 UM patients. Fourteen of these failed the genotyping or quality control, resulting in 272 UM cases available for association analysis. The vast majority of patients were ascertained from the ophthalmology, medical oncology or clinical cancer genetics clinics at Ohio State University. Four patients were referred to our research program from an outside institute. UM patients with family history of UM were excluded. The study was approved by The Ohio State University cancer institutional review board. DNA for patients and controls were extracted from peripheral blood lymphocytes by a simple salting out procedureOut of available specimens from 3500 participants currently ascertained by OSUMC Human Genetic Sample bank, for our analysis we selected 760 controls that were individually matched as closely as possible to cases by gender and age . The DNATo test the association of common genetic variants (MAF > 0.05) with the risk of UM, a total of 29 SNPs were selected through the comprehensive search of published GWAS data on melanoma risk, nevi-driven phenotypes, pigmentation, hair color, skin color and other melanoma risk etiologies. The selection criteria focused on variants with the most significant associations (in particular those surpassing GWAS level of significance) reported from each of these published GWAS. The complete list of selected variants with association information from prior studies is in For the genotyping of 29 selected variants, the highly multiplexed Sequenom MassARRAY system  was used according to the manufacturer’s protocol. Quality control (QC) measures included duplicates (8 per each 384-well plate) and non-template controls (2 per plate) resulting in >99% observed concordance with no evidence of cross-contamination. Post-genotyping filtering criteria were as follows: exclusion of SNPs with minor allele frequency (MAF) <5%, exclusion of SNPs with a call rate <95%, exclusion of samples with a call rate <95%, and exclusion of SNPs with significant departure from Hardy-Weinberg equilibrium (p < 0.001). The filtering steps excluded one variant with MAF < 0.05, resulting in genotype information on 28 variants for 272 UM patients and 735 matched controls.2 tests (1 degree of freedom). All analyses were performed in PLINK2 = 1) determined from European ancestries in the recent data release of 1000 Genomes Project Phase 3 (October 2014), as indicated in The association of UM risk with the selected 29 common genetic variants in the analysis of 272 cases and 1782 controls was estimated as odds ratios (OR) with 95% confidence intervals (95% CI), using unconditional logistic regression in an additive model where two copies of the minor allele have twice the effect of one copy. The genotype model (separate indicators for heterozygotes and rare homozygotes), dominant model (indicator for heterozygotes and rare homozygotes combined) and recessive model (indicator for homozygotes) were also assessed but not reported as they presented no significant additional information not captured by the additive model. All models were adjusted for continuous age at diagnosis (cases) or at the time of inclusion in the study (controls) and gender. Deviations of the genotype frequencies in the controls from those expected under Hardy-Weinberg equilibrium were evaluated by χIMPUTE2 was used for imputationThe imputed variants were tested for UM association using logistic regression implemented by the SNPTEST software under an additive model. Genotype uncertainties from imputation were accounted for in the model using genotype dosages. Notably, there were a number of variants comparably significant to our top genotyped associations . To asseHow to cite this article: Ferguson, R. et al. Genetic markers of pigmentation are novel risk loci for uveal melanoma. Sci. Rep.6, 31191; doi: 10.1038/srep31191 (2016).


In silico functional analysis revealed that many genes in these families might be associated with the regulation of responses to environmental stimuli via transcriptional regulation of the response genes. Moreover, these genes had diverse endogenous expression patterns in switchgrass during seed germination, vegetative growth, flower development, and seed formation. Interestingly, several members of the ERF and DREB families were found to be highly expressed in plant tissues where active lignification occurs. These results provide vital resources to select candidate genes to potentially impart tolerance to environmental stress as well as reduced recalcitrance. Overexpression of one of the ERF genes (PvERF001) in switchgrass was associated with increased biomass yield and sugar release efficiency in transgenic lines, exemplifying the potential of these TFs in the development of lignocellulosic feedstocks with improved biomass characteristics for biofuels.The APETALA2/ethylene response factor (AP2/ERF) superfamily of transcription factors (TFs) plays essential roles in the regulation of various growth and developmental programs including stress responses. Members of these TFs in other plant species have been implicated to play a role in the regulation of cell wall biosynthesis. Here, we identified a total of 207 AP2/ERF TF genes in the switchgrass genome and grouped into four gene families comprised of 25 AP2-, 121 ERF-, 55 DREB (dehydration responsive element binding)-, and 5 RAV (related to API3/VP) genes, as well as a singleton gene not fitting any of the above families. The ERF and DREB subfamilies comprised seven and four distinct groups, respectively. Analysis of exon/intron structures of switchgrass AP2/ERF genes showed high diversity in the distribution of introns in AP2 genes versus a single or no intron in most genes in the ERF and RAV families. The majority of the subfamilies or groups within it were characterized by the presence of one or more specific conserved protein motifs.  Panicum virgatum) is an outcrossing perennial C4 grass known for its vigorous growth and wide adaptability and, hence, is being developed as a candidate lignocellulosic biofuel feedstock  is a large group of regulatory protein families in plants that are characterized by the presence of one or two conserved AP2 DNA binding domains. AP2/ERF TFs are involved in the transcriptional regulation of various growth and developmental processes and responses to environmental stressors. The AP2 domain is a stretch of 60–70 conserved amino acid sequences that is essential for the activity of AP2/ERF TFs  have been suggested to be associated with the regulation of cell wall biosynthesis in some tissues Arabidopsis AP2/ERF TFs were excluded from our subsequent analysis.We used representative genes from appropriate rice gene families as the basis to search for orthologs in switchgrass. The amino acid sequences of AP2 domain-containing rice genes represented three families: AP2 (Os02g40070), ERF (Os06g40150), and RAV (Os01g04800). These proteins were used to query the derived amino acid sequences of all switchgrass AP2/ERF TFs using tblastn against the switchgrass EST database . The exon–intron organizations in these genes were visualized by the gene structure display serverThe genomic and coding DNA sequences of the identified AP2/ERF TFs were retrieved from the Phytozome (The transcript data for the AP2/ERF superfamily TFs were extracted from the publicly available switchgrass gene expression atlas (PviGEA)Arabidopsis AtSHN2 (At5g11190) and rice OsSHN (Os06g40150) was identified by tblastn or blastp against the switchgrass EST database or draft genome (Phytozome v1.1 DOE-JGI) followed by cluster and multiple sequence alignment analysis to discriminate the most closely related gene for cloning. For construction of overexpression cassette, the open reading frame (ORF) of PvERF001 was isolated from cDNA obtained from ST1 clonal genotype of ‘Alamo’ switchgrass using gene-specific primers flanking the ORF of the gene and cloned into pANIC-10A expression vector by GATEWAY recombination  and watered three times per week, including weekly nutrient supplements with 100 mg/L Peter’s 20-20-20 fertilizer. Transgenic and non-transgenic control lines were propagated from a single tiller to produce three clonal replicates for measuring growth parameters  and quantitative [pyrolysis molecular beam mass spectrometry (py-MBMS)] analysis of lignin content was performed as previously described  particle size and sugar release efficiency was determined via NREL high-throughput sugar release assays on extractives- and starch-free samples  with least significant difference (LSD) procedure was used while PROC TTEST procedure was used to examine the statistical difference between the expression of target genes in transgenic vs non-transgenic lines using SAS version 9.3 . Pearson’s correlation coefficient to determine the relationship between relative transcript levels and growth parameters was calculated by SAS.Arabidopsis along with the presence of conserved B3 domain suggest that these proteins might be categorized as putative AP2/ERF TFs. The characteristic features of these genes are summarized in Table S2 in Supplementary Material. The amino acid sequences of AP2/ERF TFs showed wide variation in size (ranging from 119 to 666 amino acids) and sequence composition. Twenty-two of these TFs contained two AP2 DNA-binding domains and hence were classified under AP2 family. Five of the AP2/ERF proteins had a B3 conserved domain at the C-terminus in addition to the common AP2 domain, and these genes were grouped into the RAV family. Three of the remaining 180 proteins, namely PvERF049, PvERF160, and PvERF177 with a single AP2 domain, which is more similar to the amino acid sequences of AP2 domains in the AP2 family TFs, were also grouped under the AP2 family. Moreover, one AP2/ERF protein showed a distinct AP2 domain different from all other switchgrass AP2/ERF proteins but with higher shared sequence similarity with the previously identified genes in rice and Arabidopsis. The remaining 176 proteins were grouped into ERF family, which was further subdivided into either one of two subfamilies (ERF and DREB) based on sequence similarity in the AP2 domain. The ERF subfamily members included 121 proteins while DREB had only 55 proteins .To confirm the classification and evaluate the sequence similarities between the switchgrass AP2/ERF TFs, a dendrogram was constructed by NJ method using the whole amino acid sequences of the proteins. The analysis showed distinct clustering of the proteins into specific groups and families as previously described in other species Figure . SpecifiTo complement the cluster analysis-based classification, the exon–intron structures of AP2/ERF genes were evaluated. The schematic representations of protein and gene structures of switchgrass AP2/ERF superfamily are presented in Figure Analysis of amino acid sequence conservation in the whole proteins of AP2/ERF superfamily showed the presence of unique conserved motifs shared between proteins within families, subfamilies, or groups Figures . MoreoveGene ontology analysis of switchgrass AP2/ERF TFs, based on rice reference sequences, predicted candidate genes’ molecular functions, putative roles in the regulation of diverse biological processes, and their cellular localization  containing expression data for about 78,000 unique transcripts in various tissues was recently developed  displayed increased expression level at radicle emergence whereas the other two (PvERF049 and PvERF203) showed increased expression at coleoptile emergence. The expression of the RAV genes and the singleton gene were apparently relatively less variable throughout the seed germination process  revealed apparent differential expression pattern between the organs and different stages of vegetative development . Moreover, the expression pattern of AP2/ERF genes during reproductive development also showed differential expression between the reproductive tissues from the initiation of inflorescence meristem to the maturation of the seeds .PvERF95, PvERF98, PvERF101, and PvERF102) and one gene in group II (PvERF148) of the DREB subfamily showed highest expression in vascular bundles and internode tissues followed by internode portions where active lignification is expected  and X  showed the highest expression in the vascular bundles followed by youngest internode sections , one gene in group VI-L (PvERF088), and three genes in group VII  had high expression in vascular bundles. Contrastingly, some genes in the ERF subfamily belonging to group V (PvERF001 and PvERF002) and VI (PvERF068) showed the highest expression in the basal fragments of the fourth internodes (E4) that is under less active lignification. Other genes including PvERF178 (VI); PvERF110 (VII), PvERF115 (VII), and PvERF164 (VII); and PvERF038 (IX) had notably high relative expression in roots than in other tissues. Compared to the ERF family genes, the expression of AP2 genes was highly diverse with some genes having high specificity to roots and vascular bundles. The expression of the two RAV genes analyzed was uniformly low throughout whereas the singleton gene was highly expressed in the LBs, LSH as well as the vascular bundles, and young internode sections  and rice OsERF057 (OsSHN) in ERF subfamily group V, for overexpression analysis in switchgrass. This gene was selected since the expression of its Arabidopsis homolog in transgenic rice resulted in modified cell wall composition  and the C-terminal motif (cm) specific to the Arabidopsis SHINE clade of TFs  and OsERF012 and OsERF057  and tiller height (R = 0.73 at P = 0.06)] . However, no tangible difference was observed in the rate of leaf chlorophyll leaching between transgenic and the control lines (data not shown). Subsequently, we analyzed whether the changes in leaf morphology might be accompanied by changes in the expression level of genes in the cutin and wax biosynthesis pathway, in which none were observed . Moreover, overexpression of PvERF001 in transgenic switchgrass showed relatively reduced expression of some lignin (PvC4H and PvPAL), hemicellulose (PvCSLS2), and cellulose (PvCESA4) biosynthetic genes, as well as some of the transcriptional regulators (PvMYB48/59 and PvNST1) of cell wall biosynthesis . The total lignin content in R1 tillers determined by Py-MBMS of cell wall residues and in leaves determined by phloroglucinol–HCl staining did not show sizeable difference between the transgenic and non-transgenic control lines . Similarly, analysis of the S/G lignin monomer ratio in transgenic lines did not significantly change as compared to that of the non-transgenic control line . However, significant improvement in glucose release efficiency was observed in lines 7 (10%) and 8 (16%) relative to the non-transgenic control line  , and Populus (66). All three species along with switchgrass have a singleton in their genome. Consistent with the previous report in rice , also known as the cytokinin responsive factor (CRF) domain in Arabidopsis that is also shared by rice ERF genes belonging to same group in rice ERF subfamily  displayed LWSY conserved motif (M21) at the C-terminal and thus may play similar roles. No information is available in the literature on some of the conserved motifs identified here including M8, M13, M14, M15, M17, and M24 , which might potentially be specific to switchgrass.AP2/ERF TFs that function as repressors or activators of specific target genes are distinguished by the presence of conserved motifs called repression domains (RD) that are highly conserved, or by the presence of activation domains which are generally less conserved , DREB , and AP2 genes  during the initiation of seed germination as well as in vascular bundles and internode sections. Moreover, the observed robust expression of 14 DREB, 17 ERF, and 3 AP2 genes in tissues or organs undergoing active lignification  but less robust expression in less lignified tissues (leaves) also supports this assertion , reduced lignification, and enhanced carbohydrate content (cellulose) in the wood of transgenic lines hinting that these TFs may indeed interact with the transcriptional machinery regulating cell wall biosynthesis  (EjAP2-1) is an indirect transcriptional repressor of lignin biosynthesis via interaction with EjMYB1 TFs  gene resulted in improved enzymatic saccharification efficiency via increased level of starch  was expressed mainly in the abscission zone and functionally associated with the regulation of the pedicel abscission zone-specific transcription of genes including cell wall-hydrolytic enzymes  required for abscission  was proposed to have a native association with cell wall regulatory and biosynthetic pathways, yet this was not experimentally verified   exhibiting reduction in plant height but increase in the number of tillers (Zhou et al., PvERF001 showing increased plant height but no difference in number of tillers. This suggests that ERF genes might functionally be highly diversified and PvERF001 may be part of a different pathway than we anticipated such as regulation of responses to biotic stress or other abiotic stress or regulation of cell elongation or division in coordination with the cytokinin pathway, with the latter perhaps explaining the observed increase in biomass and vegetative growth in transgenic lines.A recent study involving overexpression of rice homolog of PvERF001 highlight the potential of these TFs for improvement of bioenergy feedstocks.In summary, the expression profiling of the switchgrass AP2/ERF genes provides baseline information as to the putative roles of these genes and thus a useful resource for future reverse genetic studies to characterize genes for economically important bioenergy crops. With the current advancements in switchgrass research and establishment of efficient transformation system, this inventory of genes along with the information provided here could facilitate our understanding regarding the functional roles of AP2/ERF TFs in plant growth and development. Furthermore, it would aid in the identification of potential target genes that may be used to improve stress adaptation, plant productivity, and sugar release efficiency in bioenergy feedstocks such as switchgrass. The increased biomass yield and sugar release efficiency from overexpressing WW designed and performed the experiments, analyzed the data, and wrote the manuscript. MM participated in experimental design and data analysis, assisted with revisions to the manuscript and coordination of the study. GT, RS, SD, and MD assisted with performing lignin and sugar release assays and contributed in revision of the manuscript. CS conceived the study and its design and coordination, and assisted with revisions to the manuscript. All authors read and consented to the final version of the manuscript.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.http://journal.frontiersin.org/article/10.3389/fbioe.2015.00101The Supplementary Material for this article can be found online at Click here for additional data file.Click here for additional data file.Click here for additional data file.Click here for additional data file.Click here for additional data file.Click here for additional data file.Click here for additional data file.Click here for additional data file.