Exposure to lead (Pb) has been associated with endocrine, hematological, gastrointestinal, renal and neurological problems in humans. However, effects on the thyroid gland are controversial.The aim of the present study was to assess thyroid function in foundry workers occupationally exposed to Pb and the mechanism of oxidative-antioxidant imbalance.Thyroid function parameters and markers of oxidative stress were examined in 59 adult males who had been occupationally exposed to Pb. The results were then compared to those of 28 male subjects who had no history of Pb exposure or thyroid abnormalities and served as a control group.Mean blood lead levels (16.5±1.74 μg/dl) were significantly higher among the exposed workers compared to those of the control group ). The exposed group had significantly increased free triiodothyronine (FT3), free thyroxine (FT4) and significantly decreased thyroid stimulating hormone (TSH) (1.77±0.44 μIU/ml), whereas the control group had a TSH level of 2.61±0.94 μIU/ml (p< 0.0001). A state of oxidative stress was indicated by the significant increase in mean levels of malondialdehyde (MDA) and significant decrease in glutathione (GSH) (p < 0.0001). There was a significant positive correlation  between blood lead levels (BLL) and duration of employment, while BLL showed a significant negative correlation with TSH , and GSH . Of the occupationally exposed workers, 32.76% had elevated thyroid hormones. The results showed a significant positive relationship between GSH and TSH , MDA with FT3  and FT4  among exposed workers.Workers exposed to Pb dust proved to be at risk for hyperthyroidism, which was found to have a significant role in oxidative–antioxidant imbalance present among workers with increasing duration of exposure.ObtainedThis study was approved by the Ethical Committee of the National Research Centre in Egypt (NRC) under the registration number 15225.The authors declare no competing financial interests. Foundry processes involve pouring molten metal into a mold made to the external shape of the article to be cast. The mold may contain a refractory core which determines the dimensions of any internal cavity or hollow. Molten metal is then introduced into the mold. After cooling occurs, the mold is subjected to a ‘shake out' procedure which releases the casting and removes the core. The casting is then cleaned, and any extraneous metal is removed. Many changes have occurred in foundry technology and materials, but the basic processes and associated hazards have remained much the same in many foundries.1During these processes, foundry workers may be exposed to hazards such as particulate matter and metals, silica, polycyclic aromatic hydrocarbons (PAH), high temperatures and machinery.5In these environments, particulate matter is typically formed by metallic vapor condensation followed by oxidation reactions. Lead (Pb), mercury (Hg), cadmium (Cd) and zinc (Zn) are the main pollutants. Thus, foundry workers are exposed to different types of metals. Exposure to these metals results in pro-oxidant/antioxidant imbalance and can act as an intermediate in the formation of an oxidative stress state; as levels of the markers of lipid peroxidation malondialdehyde (MDA) increase and the activity of glutathione (GSH) enzyme decrease, the degree of oxidative stress could be affected.94), free thyroxine (FT4), triiodothyronine (T3), free triiodothyronine (FT3), or thyroid stimulating hormone (TSH).Occupational and chemical exposure might interfere with the hypothalamic pituitary thyroid axis at different levels and through different mechanisms of action.The present work was a cross sectional comparative study conducted in a non-ferrous foundry plant in Helwan, Cairo, Egypt. The foundry plant manufactures aluminum, Pb, Zn, copper and precious metal products.BLLBlood lead level3FTFree triiodothyronine4FTFree thyroxineGSHGlutathioneMDAMalondialdehydeROSReactive oxygen species3TTriiodothyronine4TThyroxineTSHThyroid stimulating hormoneThe present study was conducted from April 2016 to May 2017.Supplemental Material.The study population was comprised of the work force in the Pb casting departments. After applying the exclusion criteria, which included workers who have undergone thyroid surgery or receiving any form of thyroid treatment, 61 workers were eligible for inclusion and only two workers did not agree to participate in the study. Another group of referents (n=28) were randomly selected from men employed at administrative jobs who lived in residential areas away from the factory and were never occupationally exposed to metals. Both groups were matched for age, social economic status and smoking habits. All subjects were interviewed using a questionnaire involving occupational history, and clinical examination, including thyroid gland inspection and palpation, was performed by a specialized physician. The questionnaire can be found in This study was approved by the Ethics Committee of the National Research Centre in Egypt (NRC) under the registration number 15225.Blood samples were collected from all subjects using a dry plastic disposable syringe and divided into two parts: the first part (3 ml) was collected into K-EDTA tubes for Pb and GSH evaluation, the second part (3 ml) was collected into serum vacationer tubes for thyroid hormones and MDA measurements.et al.17Lead level evaluation was performed using the simultaneous inductively coupled plasma emission spectrometer (Agilent 720 ICP-OES) and the method described by Momen 3, FT4 and TSH were carried out using an enzyme immunoassay kit purchased from International Immuno-Diagnostics Co., USA (Gamma Trade Company), and the methods described by Melmed et al., Tarnoky, and Synder et al.20Quantitative measurements of serum FTet al.et al.22Quantitative determination of serum MDA was carried out calorimetrically using a kit purchased from Biodiagnostic Co., Egypt, according to the method described by Ohkawa P-values <0.05 were considered statistically significant. Multiple linear regression analysis was used to estimate the influence of independent variables such as GSH and MDA on the markers studied (dependent variables).Statistical evaluation of all results was conducted using the Statistical Package for the Social Sciences software (SPSS) version 16. The mean values, SDs and ranges were estimated for quantitative variables. The correlations between individual variables were calculated using Pearson correlation coefficient. p <0.0001). Their mean duration of employment was 22.35±10.7 years. The exposed group showed a significant increase in thyroid hormones  compared to the control group (p <0.0001). In addition, TSH mean value (1.77±0.44 μIU/ml) was significantly decreased compared the control group (2.61±0.94 μIU/ml). The exposed group also showed a state of oxidative stress represented by a significant increase in mean levels of MDA and a significant decrease in mean levels of GSH (p <0.0001) .The exposed group had an age range of 28–59 years with a mean value of 44.9±10.36 years, which did not differ significantly compared to the control group. Smoking habits showed no significant difference between the two study groups. Mean blood lead levels (BLL) were significantly higher among the exposed workers (16.5±1.74 μg/dl) compared to the control group (12.8±1.16 μg/dl) (p <0.05) and a negative correlation with TSH , and GSH  .In the correlation of BLL with duration of employment, there was a statistically significant positive correlation . Exposed workers with elevated thyroid hormones represented 32.76% of exposed workers .Using the independent t-test to compare the mean value of oxidative stress biomarkers between exposed workers with normal FT3 and FT4 as independent variables. The results showed a significant positive relationship between GSH and TSH, and between MDA and FT3 and FT4 among exposed workers .Multiple linear regression analysis was applied with MDA or GSH as dependent variables, using TSH, FTOccupational exposure to heavy metals can cause many harmful health effects, depending on the intensity and duration of exposure. Many studies have suggested that occupational and environmental exposure to heavy metals such as Hg, Cd, chromium, arsenic, nickel and Pb cause oxidative damage and are capable of disrupting the activity of several proteins in the reproductive and endocrine system.27Oxidative stress is a well-documented mechanism of metal toxicity and carcinogenicity. It is the result of imbalance between radical oxygen species production and the antioxidant defense system. Redox-inactive toxic metals such as Pb deplete cells of antioxidant reserves, especially GSH, which plays a pivotal role in its overall toxic manifestations.28p <0.05), which indicates that work activity had a direct relationship with the risk of occupational exposure to Pb and indicates an increased body burden of Pb among the exposed workers due to their occupational setting.The characteristics of the study population are summarized in Some metals have the ability to produce reactive oxygen species (ROS) in biological systems, which may lead to a state of oxidative stress. It is a state where increased formation of ROS overwhelms the body's antioxidant production and subsequently induces lipid peroxidation, DNA damage, protein modification and other effects. The underlying mechanism of these toxic metals involves the production of MDA and depletion of GSH. Lead may induce oxidative stress that may deteriorate biological macromolecules either by increased ROS or depletion of a cell's major antioxidants.30et al. and Sciskalska et al.3 and FT4 mean levels compared to the control group. Lead is known to have adverse neurological, hematological, renal and gastrointestinal effects. However, associations with thyroid hormones have been inconsistent.14The present study found that the exposed group suffered from a state of oxidative stress, indicated by significantly higher serum MDA and a significant lower GSH in the exposed group compared to the controls. The higher MDA level is consistent with the results of Liu et al. found that FT3 levels were significantly higher in subjects with Pb exposure compared to the control group (p <0.01), TSH levels were lower (p < 0.001), but no significant difference was found for FT4 between the two groups.4 and non-significant reduction of TSH in Pb-exposed workers compared with controls.3 and T4.3 and FT4.15Yilmaz Lead is a redox-inactive metal, it depletes cells' major antioxidant reserves, especially GSH, which plays a pivotal role in its overall toxic manifestations.37p <0.05). Our results are in agreement with those of many studies which suggests that Pb is a redox inactive metal that induces oxidative stress in cells and can be partially responsible for its toxicity.40The disulphide bond is found in the active site of glutathione reductase, the disulphide bond interferes with Pb and inhibits enzymes. The inhibition prevents glutathione disulfide from being reduced to GSH, thus Pb deplete cells' major antioxidants, enhanced generation of ROS and results in an oxidative stress state.p <0.001) and no association with FT3 and FT4, although they were significantly higher in the exposed group compared to the control. Previous studies have suggested the same association among males and females, while others found negative associations with FT3 and FT4, however, associations were not evident in other similar studies.46The present study reported a negative association between BLL and TSH .3 and FT4 as independent variables. A positive association between GSH and TSH , and MDA with FT3  and FT4  was observed among exposed workers.Paint workers exposed to Pb and solvents were at risk for hyperthyroidism, as TThe results of the present study suggest that occupational exposure to Pb dust and fumes has a stimulatory effect on thyroid function as manifested by a significant increase in thyroid hormone levels, even if this increase was not associated with clinical manifestations of hyperthyroidism. Foundry workers exhibited an increase in MDA levels and decrease in GSH levels, which represents evidence for oxidative stress imbalance, in which increased thyroid hormones play a significant role in ROS production through stimulation of metabolism and the increase of MDA levels. In addition, exposure to Pb, a redox inactive metal, depletes cells' major antioxidants reserves of GSH. Mounting evidence indicates that multiple mechanisms may be responsible for the oxidative stress imbalance caused by exposure to toxic metals such as Pb, including thyroid stimulation.Click here for additional data file.


Pseudomonas aeruginosa clinical isolates and identified single-nucleotide polymorphisms that are the basis for branching of the phylogenetic tree. We also identified single-nucleotide polymorphisms that were acquired independently, in separate lineages, and not through inheritance from a common ancestor. Although our results demonstrate that the Pseudomonas aeruginosa core genome is highly conserved and in general, not subject to adaptive evolution, instances of parallel evolution will provide an opportunity to uncover genetic changes that underlie phenotypic diversity.Extensive use of next-generation sequencing has the potential to transform our knowledge on how genomic variation within bacterial species impacts phenotypic versatility. Because different environments have unique selection pressures, they drive divergent evolution. However, there is also parallel or convergent evolution of traits in independent bacterial isolates inhabiting similar environments. The application of tools to describe population-wide genomic diversity provides an opportunity to measure the predictability of genetic changes underlying adaptation. Here, we describe patterns of sequence variations in the core genome among 99 individual  A major challenge in biological and environmental research is to understand the intricate link between genotypes and phenotypes and how the environment influences that link. Genetic variants such as mutations or the presence or absence of a gene among individuals can produce different phenotypes that undergo positive or negative selection. Thus, evolutionary change critically relies on genetic variations. The genetic variation within a species is often called genetic diversity and typically corresponds to phenotypic variation. In a genetically diverse population, it is more likely that at least some individuals survive in changing environments .Pseudomonas aeruginosa represents an excellent model for understanding the molecular mechanisms of the adaptation of a bacterial species to a variety of challenging habitats. The species P. aeruginosa inhabits a stable source habitat in the natural environment, which serves as a reservoir to cause infectious diseases in humans occasionally. If not eradicated during acute infection, P. aeruginosa populations can cause chronic infections that exist for a long enough time for evolution to take place . Our analysis also uncovered convergent adaptation signatures that evolved independently of the phylogenetic background of the clinical isolates. Analysis of the strict core genes of a collection of 99 clinical isolates furthermore revealed that the P. aeruginosa core genome is highly conserved across clinical isolates and is generally not subject to adaptive evolution.Based on annotated genes across a set of 99 genotypic diverse clinical P. aeruginosa that were sampled from various infection sites at different hospitals across Germany. Only one isolate per patient was included in this study. Genomic DNA was prepared from P. aeruginosa isolates using the NEBNext Ultra Kit and sequenced on an Illumina MiSeq System, generating 2× 300-bp paired-end reads. Using a multiplexed protocol, an average of 1,037,171 reads  for each of the genomic libraries were obtained. On average, the isolates were sequenced with an estimated genomic coverage of 68-fold   ; Karect 4, 2020) . The pro4, 2020) . Its bui4, 2020) , was use4, 2020) .P. aeruginosa PA14 and PAO1 strain types were downloaded from GenBank ], to the data  to the rate of synonymous substitutions (dS) was calculated based on the modified Nei and Gojobori method  under accession number SRP160898.All short-read data are available at the National Center for Biotechnology Information Sequence Read Archive  per individual isolate (details in lasR exhibited (incomplete) gene losses in more than 5% of our clinical isolates, and therefore was not considered as soft core, but as an accessory gene in our analysis. Thus, our total of 1,257 soft-core genes is only an estimate that could be further curated as more isolates are sequenced.Whole-genome sequencing of a multitude of bacterial isolates from one species gives detailed information on the available gene pool. This approach provides information on the number of genes that are common to all isolates (core genome), the number of genes that are found in a subset of strains (accessory genome), and those that only occur in a single strain (singletons) . However Germany  online. P. aeruginosa pan-genome were comparable to those previously published. We found a large number of strict  and soft  core genes. This was in line with previous reports on a high degree of conservation between P. aeruginosa isolates  isolates. It was estimated that the pan-genome is saturated with 25,946 genes and 940 genomes would provide a 95% saturation. Saturation was also estimated with 3,050 strict core genes (683 genomes for 95% saturation), 7,076 singleton genes and 18,475 soft singletons . Furthermore, a saturated accessory genome size of 8,061 genes was estimated   B. The fik-mers of the 3,814 core genes. The core genome, which includes the seven standard multilocus sequence type (MLST) genes of P. aeruginosa   , represeectively  fig. 3)k-mers of strains  online.P. aeruginosa core genes is provided in the Bactome database    and exemP. aeruginosa genomes amounting to overall 159,609 single-nucleotide polymorphisms (SNPs). This corresponds to a mean sequence diversity at the single-nucleotide level of 0.04 , indicating that the core genes are highly conserved within these 99 strains  were scanned for sequence variations across the 99  strains . Seven gAs expected, the sequence variation among the isolates was not random, but isolates that belonged to the same phylogroup shared patterns of SNP profiles . The PA1In addition to the 75,765 phylogroup/divergent SNPs and the 49,722 single SNPs (which occurred only in one isolate), a total of 34,122 SNPs were identified across many isolates, independent of the phylogenetic background . Of these 34,122 SNPs, 27,590 SNPs (∼80%) were identified as synonymous, whereas 6,532 (∼ 20%) were identified as nonsynonymous. Those SNPs were found in 3,497 and 2,252 genes, respectively . A list We used the 75,765 SNPs that are divergent in the population to construct a phylogenetic tree. In comparison to the original tree based on all core genome SNPs, the tree shows a slight variation in distances across and within phylogroups and depicts clear sorting within phylogroup nodes, especially in PA14 group  online.N) to synonymous substitutions rates (dS), dN/dS, remains one of the most common measures used to describe stabilizing selection. A ratio of dN/dS > 1 indicates positive selection, a dN/dS ratio of 1 corresponds to neutral selection, and dN/dS < 1 denotes a purifying (or negative) selection  exhibited a dN/dS ratio between 1 and 2. In an extended analysis, we determined the dN/dS ratio for the genes present in 2–5 isolates . The dN/dS ratio of 0.06 indicates that there is no positive selection on these genes . Recently, two additional taxonomy outliers with the PA7 phylogroup were identified  and aligned the orthologous sequences of all individual core genes. Any sequence variation at any position within the aligned genes gave information on SNPs at high accuracy.Here, to first gain more information on the structure of the global gene pool within our studied -genomes . Of noteentified . We thenIn the last decade, MLST has been used to study the molecular epidemiology of bacterial pathogens . MLST meP. aeruginosa isolates paved the way for categorizing SNPs that define the branches of a phylogenetic tree (divergent SNPs), and SNPs that have been acquired independently in separate lineages, and not through inheritance from a common ancestor (convergent SNPs). We identified nearly 47.47% of the total SNPs as phylogroup dependent (divergent), whereas 21.38% were commonly found independently of the strain background (convergent). Approximately, 31.15% of the total SNPs were singleton SNPs with an average of 492 singletons per isolate. A large fraction of these singletons were identified in hypermutator strains, which exhibited a high mutation frequency due to mutations in DNA repair systems. For example, CH3570, CH2922, MHH15286, CH2824, CH2804, and CH2677 harbored >2,000 singleton SNPs, which is more than 4-fold than the average number of singletons per isolate. However, also both PA14 and PAO1 contained an above average numbers of singletons—811 and 1,118, respectively. This indicates that the two lab strains have deviated quite substantially from natural P. aeruginosa isolates. The categorization of SNPs that define the branches of a phylogenetic tree (divergent SNPs) in the 99 clinical P. aeruginosa isolates across the entirety of the core genes provided the basis for unprecedented detail into the phylogenetic relatedness between the individual isolates of the P. aeruginosa population.Detailed knowledge on the full pattern of sequence variations among the core genes of the 99 clinical N) to the synonymous substitutions rate (dS), dN/dS for the 3,814 core genes to be 0.14. The application of dN/dS methods can be challenging when analyzing within-species data, which represent mutations not yet fixed in the population (P. aeruginosa core genome is, for the most part, not subject to adaptive evolution.It has been shown previously that the core genes are highly conserved . In accopulation . NeverthP. aeruginosa (P. aeruginosa determines environmental adaptability such as niche expansion (P. aeruginosa (Previous comparative genomic studies have demonstrated that the core genes of bacteria play important roles in niche adaptation and virulence, especially in ruginosa . Althougruginosa , and it xpansion . A recenruginosa . Furtherruginosa . BecauseP. aeruginosa isolates analyzed in this study is conserved and is not subject to positive selection. The future analysis of full genomes that include soft-core, accessory, and intergenic regions will allow for more detailed information on the structure and dynamics of the P. aeruginosa SNP profile and how this contributes to bacterial phenotypes.In conclusion, we have shown that the core genome of the 99 clinical Genome Biology and Evolution online.evaa059_Supplementary_DataClick here for additional data file.


Calcium and fluoride ion releases were evaluated. Biofilms of human saliva were assessed. (3) Results: nCaF2+DMAHDM+MPC composite had the lowest biofilm colony forming units (CFU) and the greatest ion release; however, its mechanical properties were lower than commercial control composite (p < 0.05). nCaF2+DMAHDM composite had similarly potent biofilm reduction, with mechanical properties matching commercial control composite (p > 0.05). Fluoride and calcium ion releases from nCaF2+DMAHDM were much more than commercial composite. Biofilm CFU on composite was reduced by 4 logs . Biofilm metabolic activity and lactic acid were also substantially reduced by nCaF2+DMAHDM, compared to commercial control composite (p < 0.05). (4) Conclusions: The novel nanocomposite nCaF2+DMAHDM achieved strong antibacterial and ion release capabilities, without compromising the mechanical properties. This bioactive nanocomposite is promising to reduce biofilm acid production, inhibit recurrent caries, and increase restoration longevity.(1) Background: The objective of this study was to develop a novel dental nanocomposite containing dimethylaminohexadecyl methacrylate (DMAHDM), 2-methacryloyloxyethyl phosphorylcholine (MPC), and nanoparticles of calcium fluoride (nCaF Dental resin composites are an excellent material for direct restorations of anterior teeth and in many cases posterior teeth due to their esthetics and ease of placement . Neverth2) into composites has the potential to reduce demineralization [5(PO4)3F] to protect the tooth surface ), where load P was divided by displacement d which is the slope in the linear elastic region. Six specimens were tested for each group (n = 6).Each composite paste was mixed in a disposable plastic container using a speed mixer  at a speed of 2800 rpm for 1 min, and then thoroughly mixed by hand on a plastic slab for 5 min. The paste was then placed in a rectangular mold of 2 × 2 × 25 mmor 1 min . After dor 1 min ,37. Flex2 were tested. A solution of sodium chloride (NaCl) (133 mmol/L) was buffered with 50 mmol/L HEPES to pH 7   . FurtherF2 (23%) . Groups 2+DMAHDM substantially lowered the biofilm development and production of lactic acid, reducing the CFU of biofilm by 4 logs.Recurrent caries is considered a major drawback of composite restorations ,52,53. S2 and MPC exhibited a greater lactic acid production when compared with the other groups. DMAHDM has been shown in previous work to have a strong antibacterial effect, as illustrated by the CFU data in S mutans biofilm accumulates, the secretion of lactic acid occurs. In the absence of a viable biofilm, the concentration of lactic acid is expected to be very low, as is the case in the DMAHDM-containing groups. Interestingly, the nCaF2 and nCaF2+MPC groups did not have a significant reduction in CFU  influenced the hydrogen bond network and the diffusion through it [− ions would have a similar result, which may affect F ion diffusion and disrupt the creation of a protein-repellant coating, leading to biofilm formation.It can be seen in n in CFU , but theoduction . It has oduction . Howeverrough it . It is p2, and nCaF2+MPC groups exhibited the formation of microcolonies and early biofilm multilayers. The biofilms on these groups at 96 h were fully mature, substantial, and dense. However, in DMAHDM-containing groups there was minimal attachment of biofilm observed. At 96 h the DMAHDM-containing groups showed a reduction in biofilm attachment compared to earlier time points. It is possible that the antimicrobial effect of DMAHDM as a contact-killing agent had eliminated the bacteria left on the samples over time. These results correlate very well with the CFU results.SEM images in It is possible that the salivary proteins adsorption on composite resin surfaces might reduce the effectiveness of this “contact-killing” mechanism. Therefore, a protein repellent agent was added. MPC is a methacrylate that contains phospholipid polar groups. MPC is highly hydrophilic and has been shown to decrease the adsorption of protein and reduce bacterial attachments . Previou2 and was further substantially improved with the use of both nCaF2 and DMAHDM. However, the nCaF2+MPC group showed no reduction in the CFU counts. The composite that had nCaF2+DMAHDM+MPC displayed the most potent antibacterial effect with CFU 6 log biofilm reduction. Therefore, these results confirmed that the addition of DMAHDM is vital to improving the antibacterial effect. It is thought that DMAHDM serves to reduce the accumulation of biofilm and, as a result, improves the efficacy of MPC in resisting protein adsorption. However, in the absence of DMAHDM, the bacterial biofilm will accumulate and render MPC less effective.In the current study, the antibacterial capability of the experimental composite was improved with nCaF2+DMAHDM promoted remineralization and had a significant antibacterial effect. Another potential benefit of using nCaF2 is that the release of F ions could act as a transmembrane proton carrier and an inhibitor to the glycolytic enzyme, thus preventing oral microorganisms by stimulating acidification of the cytoplasm and providing antibacterial effect at a long-distance to inhibit caries [2+DMAHDM composite exhibits excellent mechanical properties which make it suitable in a variety of restoration applications. In comparison, the composite consisting of nCaF2+DMAHDM+MPC exhibited lower mechanical properties. However, the strength and modulus achieved by this group was still high enough to be used for restorations in low load areas such as class V restorations [2+DMAHDM combination in dental cements, bonding agents, fissure sealants, and composites to remineralize tooth lesions and suppress biofilm and plaque buildup, especially in patients with a high caries risk.The combination of nCaFt caries ,63. Thesorations . This is2+DMAHDM for the first time. Release of F and Ca ions from the composite was significantly achieved through the incorporation of nCaF2. The new composite exhibited strong antibacterial and ion release capabilities, prevented biofilm production of lactic acid, and decreased biofilm CFU by 4 log. This bioactive nanocomposite is promising to protect tooth structures, inhibit demineralization, and provide a reservoir for the release of calcium and fluoride ions. The combined effect of nCaF2+DMAHDM shows promise in a variety of dental applications where remineralization and prevention of recurrent caries is a priority.A novel composite with remineralization and antibacterial properties was established by combining nCaF


To evaluate the role of viral infections in the pathogenesis of primary acquired nasolacrimal duct obstruction.The study included 48 patients diagnosed with primary acquired nasolacrimal duct obstruction undergoing dacryocystorhinostomy surgery. Prior to dacryocystorhinostomy surgery, nasal swab sample was taken from the inferior meatus at the same side. During dacryocystorhinostomy, tissue biopsy sample (2 × 2 mm) was taken from the junction area of the lacrimal sac and nasolacrimal duct. Following nucleic acid extraction, polymerase chain reaction was performed.p = 0.001).The patients consisted of 9 (18.8%) men and 39 (81.2%) women with a mean age of 51.0 ± 14.3 years. Qualitative polymerase chain reaction showed viral genome in the nasal swabs of 10 (20.8%) patients, including coronavirus 229E (three cases), coronavirus HKU1 (two cases), respiratory syncytial virus (two cases), coronavirus OC43 (one case), coronavirus NL63 (one case), and adenovirus (one case). In the dacryocystorhinostomy samples, viral genomes were detected in four (8.3%) cases, including respiratory syncytial virus (two cases), coronavirus HKU1 (one case), and adenovirus (one case). There was a statistically significant agreement between nasal mucosal swab and dacryocystorhinostomy biopsy samples in terms of respiratory syncytial virus positivity (kappa = 1.000, Although the viral genome was detected in the samples, a direct relationship between viruses and pathogenesis of primary acquired nasolacrimal duct obstruction could not be revealed because of the low number of positive results. However, considering the profibrotic characteristics of specific viruses such as respiratory syncytial virus and adenovirus, viral infections may be one of the many predisposing factors of primary acquired nasolacrimal duct obstruction. Previous histopathological studies have revealed chronic inflammation of the nasolacrimal duct and the development of secondary fibrosis on the ground of this inflammation.24Primary acquired nasolacrimal duct obstruction (PANDO) is the most common cause of epiphora in adults.5 In one experimental model of dacryocystitis, the number of collagen fibers and fibrocytes in the lamina propria had increased 3 months after a Staphylococcus aureus inoculation.6 Viruses, as superior to bacteria, have capability to extend beyond the epithelium to involve the elastic tissue of the lamina propria, causing inflammation and cicatricial changes in the mucosa.5PANDO has been reported to be able to occur secondary to unrecognized low-grade dacryocystitis.710 A similar mechanism may be possible for the pathogenesis of PANDO which has common histopathological characteristics with these diseases.Recently, viral infections have been reported as the possible trigger factors in the pathogenesis of several diseases, such as idiopathic pulmonary fibrosis, dilated cardiomyopathy, and arrhythmogenic right ventricular dysplasia, characterized by inflammation and secondary fibrosis in their histopathological examinations.11 In addition, respiratory mucosa and nasolacrimal duct has the same sialic acid sequences in the end of the glycoproteins that serve almost as a receptor for infectious agents.12 These close anatomical and immunological relationships may cause an expansion of the virus inoculation to the nasolacrimal duct during upper respiratory tract infections. Similarly, viral conjunctivitis may easily extend into the lacrimal sac and nasolacrimal duct mucosa in the direction of tear drainage. A higher frequency of histories of infectious  conjunctivitis in cases with PANDO was previously reported.13 The aim of the present study was to evaluate the role of respiratory and ocular viruses in the PANDO pathogenesis.The respiratory mucosa and nasolacrimal duct epithelium has similar immunological and histological features. First, the nasolacrimal duct is a direct extension of the nasal mucosa and has been shown to include ectopic nasal epithelial cells.This prospective study included 48 patients diagnosed with symptomatic PANDO who underwent dacryocystorhinostomy (DCR) surgeries. Approval from the ethics committee was obtained, and the study was performed in adherence to the tenets of the Declaration of Helsinki. Informed consent was obtained from each of the participants prior to enrollment.All the patients underwent complete ocular and periocular examinations. The PANDO diagnosis was confirmed via lacrimal system probing and irrigation. All factors that could cause the secondary acquired nasolacrimal duct obstruction were questioned and evaluated in detail. Also, all patients underwent otorhinolaryngology consultation to rule out intranasal mass lesions. Any cases with suspected or positive history and/or findings in favor of other causes of nasolacrimal duct obstruction and cases with intranasal mass lesions were excluded. Those patients with histories of viral conjunctivitis or upper respiratory tract infections within the previous 3 months were excluded. In addition, those patients with ocular diseases that could have affected the lacrimal drainage system were also excluded.Prior to the DCR surgery, a nasal mucosal swab sample was taken from the same side of the inferior meatus of each patient. During the DCR surgery, tissue biopsy samples (2 × 2 mm) were taken from the junction area of the lacrimal sac and the proximal nasolacrimal duct. The samples were immediately transferred to the Department of Medical Microbiology under cold chain.The viral genome extraction was performed with an EZ1 Virus Mini Kit v2.0  using an EZ1 Advanced XL Robotic Workstation (Qiagen) according to the manufacturer’s instructions. After performing the extraction, the samples were frozen at −80°C.® Q ” device in accordance with the manufacturer’s recommendations. FTD Respiratory 21 kit includes influenza A virus, influenza B virus, influenza A (H1N1) virus, human rhinovirus, human coronavirus NL63, human coronavirus 229E, human coronavirus OC43, human coronavirus HKU1, human parainfluenza 1, human parainfluenza 2, human parainfluenza 3, human parainfluenza 4, human metapneumoviruses A/B, human bocavirus, Mycoplasma pneumoniae, human respiratory syncytial viruses (RSVs) A/B, human adenovirus, enterovirus, human parechovirus, and internal control. FTD Eye kit includes herpes simplex virus 1, herpes simplex virus 2, varicella zoster virus, Chlamydia trachomatis, human adenovirus, and internal control. Reactions which were in the desired cycle threshold and had exponential phase were regarded as positive.Polymerase chain reaction (PCR) using real-time technology for qualitative analysis of the pathogens was performed with 5-tube multiplex PCR kit “FTD Respiratory 21 ” and 2-tube multiplex PCR kit “FTD Eye” on the “Rotor-Genep < 0.05 was considered statistically significant.The statistical analysis was performed using SPSS software for Windows version 15.0 . Descriptive statistics were expressed as mean ± standard deviation or median (minimum–maximum) for continuous variables and as the number of observations and percentage for categorical variables. Differences in categorical variables were assessed by Pearson’s chi-square test or Fisher’s exact test. Statistical agreement between virus positivity in DCR biopsy specimens and nasal swab specimens was tested by the kappa coefficient (κ). A value of p = 0.002). The patients’ demographic data are summarized in p = 0.008).The study subjects consisted of 9 (18.8%) males and 39 (81.2%) females with a mean age of 51.0 ± 14.3 years. The female-to-male ratio was 4.3. The mean age of the females was 48.0 ± 13.4 years, and the mean age of the males was 63.7 ± 11.2 years. The mean age difference between the two genders was statistically significant  of the cases. With regard to the unilateral cases, 18 (37.5%) patients had right-sided involvement and 13 (27.1%) had left-sided involvement. Epiphora was the most frequent clinical finding that was seen in all the cases. The other examination findings related to the lacrimal drainage system obstruction were punctual regurgitation with sac compression in 31 cases (64.6%) and localized swelling in 13 cases (27.1%).p = 0.187) . The RSV was discovered in both the nasal mucosa and DCR biopsy samples in two patients (4.2%), and there was statistically significant agreement between the nasal mucosa and DCR biopsy samples in terms of the RSV positivity . In the other two patients in whom a virus (human coronavirus HKU1 in one and a human adenovirus in the other) was detected in the DCR biopsy sample, the virus was not detected in the nasal mucosa.The PCR results showed the presence of viral genome in the nasal swab samples of 10 (20.8%) of the patients. The viruses that were detected included human coronavirus 229E in three cases, human coronavirus HKU1 in two cases, human RSV in two cases, human coronavirus OC43 in one case, human coronavirus NL63 in one case, and a human adenovirus in one case. In the DCR biopsy samples, viral genomes were isolated in four (8.3%) of the cases, including human RSV in two cases, human coronavirus HKU1 in one case, and a human adenovirus in one case. The demographic characteristics of the cases in which viral genome was detected and the side with the viral genome are summarized in = 0.187) . In addi3 The exact mechanisms leading to chronic inflammation and secondary fibrosis, which are the predominant findings of the PANDO histopathology,24 have not yet been elucidated. The only previous study that evaluated the viral presence in PANDO was performed by Kashkouli et al.14 They examined the biopsy specimens of 51 PANDO cases for the presence of herpes simplex virus 1, herpes simplex virus 2, and human papillomavirus. However, the immunohistochemistry results were negative for the viruses in all the cases. To the best of our knowledge, the present study was the first to report the presence of respiratory viral genomes directly in the proximal nasolacrimal duct mucosa of four patients with PANDO.PANDO is the most common cause of acquired nasolacrimal duct obstruction in the adult.11 speculated that ascending infection of ectopic nasal epithelial cells in the nasolacrimal duct mucosa during nasal inflammation could be the starting point of dacryostenosis. Then, this inflammation may initiate dysfunction of the cavernous body with reactive hyperemia, edema of the mucous membrane, and temporary occlusion of the lacrimal passage. Repeated episodes of dacryocystitis may end up with the structural epithelial and subepithelial changes. Considering the fact that acute viral respiratory tract infections are the most common illnesses in all individuals, repetitive inflammation caused by the nasal infection may be direct or indirect stimuli for the nasolacrimal duct obstruction, especially in predisposed individuals.Paulsen et al.15 It has the capability of affecting the cytokine production profile of adaptive immune system cells by shifting the T helper (Th) 1 / Th 2 balance toward Th 2 cytokines and immunoglobulin E.16 It has also been shown to alter the biological activity of macrophages.17 Furthermore, matrix metalloproteinase 9 (MMP-9), a profibrotic enzyme has been shown to increase in the epithelium after an RSV infection.18 When all these mechanisms are considered together, RSV has the capability to trigger inflammation and fibrosis development by both shifting the Th balance toward the production of the profibrotic cytokines, interleukin (IL)-4 and IL-13, and increasing the synthesis of connective tissue enzymes, such as MMP-9.19In the present study, two of the patients exhibited the RSV genome in both the nasal mucosal swab and the DCR biopsy specimens. RSV is a segmental, negatively polarized RNA virus.2022 Early region 1A, which is a viral gene product of adenoviruses, has been reported to promote the production of profibrotic mediators, such as connective tissue growth factor and transforming growth factor-β mRNA on the bronchial epithelium.23 In the case of a latent adenovirus infection, the continuous release of these profibrotic mediators may trigger fibrosis development. In addition, the nasolacrimal duct is surrounded by a mucosa-associated lymphoid tissue, which is believed to provide immunomodulation, and damage to this structure has been associated with the development of PANDO.25 Because adenoviruses have been proven to cause latent infections in lymphoid tissues,22 a persistent viral infection in this special lymphoid tissue may lead to a chronic inflammatory process resulting in injury and scar formation.Adenovirus DNA was detected in the DCR biopsy specimen from one case and in the nasal swab sample from one case. Adenoviruses, which are common respiratory tract viruses with more than 50 serotypes, have been proven to cause latent infections, especially in lymphoid tissues.26 the human coronavirus HKU1 detected in the DCR biopsy material may be a case of an asymptomatic infection through the ocular surface. In our study, the seven patients (14.6%) with coronaviruses in their respiratory tracts were believed to be asymptomatically infected cases.One case exhibited human coronavirus HKU1 RNA in the DCR biopsy sample. In this case, no virus was detected in the nasal swab. When considering the fact that the ocular mucosa is one of the coronavirus transmission routes,27 Therefore, further studies including large number of cases with equal gender and age distribution and usage of quantitative real-time PCR analysis for suspected viruses to decide the clinical stage of the infection are needed. Animal models in which virus inoculation is performed directly into the lacrimal sac may also help to assess the mucosal changes that viruses can reveal in this region. Moreover, population-based studies with a long follow-up time that evaluate the time period prior to the disease development may help to elucidate the precise pathogenesis of PANDO.This was a pilot study, and the major study limitations were the small sample size and the low number of viral positives in the DCR samples, which prevented us from establishing a definitive association between PANDO and viral infections. Sexual differences in cytokine responses to infection may also have a role in determining susceptibility to viruses.In conclusion, the predominant findings in the PANDO histopathology are inflammation and secondary fibrosis, but the factors that may trigger the critical process have not yet been elucidated. Many inflammatory, profibrotic, and immunological pathways may have roles in the pathogenesis of PANDO. In the present study, although the viral genome was detected in the DCR samples, due to the low number of positive results, a direct relationship between the presence of viruses and the PANDO pathogenesis could not be established. However, considering the profibrotic characteristics of specific viruses such as RSV and adenovirus, viral infections may be one of the many predisposing factors of PANDO.


Cow variability present in nutrient digestibility studies differs for different diets and nutrients. It is a major factor determining adequate sample size so that studies are not under-powered or over-powered. The objective of the current study was to develop cow variability estimates that can be used to determine the optimal sample size for digestibility trials having randomized block designs using mid-lactation dairy cows when fed corn-based diets having different neutral detergent fiber:starch ratio . Cow variability is greater for digestibility of fiber and dry matter and less for starch. Estimated cow variability as standard deviations for digestibility of dry matter, neutral detergent fiber and starch were 3.8 g/kg, 5.1 g/kg and 3.3 g/kg, respectively. A major implication of this study is that cow variability is greatest for fiber digestibility and the use of a minimum of 12 cows per dietary treatment is adequate to reliably detect treatment effects on the digestibility of fiber, starch and dry matter using lactating dairy cows fed in groups with randomized block design under current experimental conditions. p > 0.05). Estimated cow variability (as standard deviation) for digestibility of dry matter, NDF and starch were 3.8 g/kg, 5.1 g/kg and 3.3 g/kg, respectively. A major implication of this study is that cow variability is greatest for NDF digestibility and the use of a minimum of 12 cows per dietary treatment is adequate to reliably detect treatment effects on the digestibility of NDF, starch and dry matter using cows fed in groups with randomized block design under these experimental conditions. The objective of this study was to estimate cow variability that can be used to determine the optimal sample size for digestibility trials using lactating dairy cows. Experimental design was randomized complete block design having three blocks and three dietary treatments. Three similarly managed nearby intensive farms were considered as blocks, and three diets were formulated to have 0.7, 1.0, and 1.3 neutral detergent fiber (NDF): starch ratio. In each farm, 18 cows were assigned for each dietary treatment and five sample sizes per each treatment group were simulated by simple random sampling of data from 18, 15, 12, 9 and 6 cows respectively. Intake was not affected by diet or sample size ( The gold standard for determining nutrient digestibility in dairy cows is direct measurement under production conditions and is not completely replaceable by metrics such as in vitro or in situ procedures . HoweverPower analysis is the most scientifically favored approach to determine the appropriate sample size . The sizFor dairy cow nutrient digestibility experiments with completely randomized designs, few between-cow variability estimates for dry matter digestibility (DMD), NDF digestibility (NDFD), and starch digestibility (STRD) are reported in the literature. In one study, between-cow variation for DMD was higher SD = 4.24 g/kg) than NDFD (SD = 2.82 g/kg) when cows fed diets with or without yeast (Saccharomyces cerevisiae) fermented-cassava chips when the experiment was designed in agreement with randomized complete block design (RCBD) . In anot4 g/kg thAll experimental procedures and animal use protocols were approved by the Animal Care and Use Committee of the Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China (No. IAS20180115). The experimental design was a randomized complete block design. A digestibility trial was composed of three blocks and three dietary treatments. Three similarly managed intensive dairy farms (500 to 5000 cows) within close proximity of each other in northern China were used and considered as blocks . Three eThe sample size for the feed trial was projected at the planning stage using PROC MIXED procedure of SAS  using the PARMS statement. Anticipated mean NDF digestibility for three treatment groups , α = 0.05 and power = 0.80 were used to determine the appropriate sample size. Cow within a block, pen and treatment was considered as the experimental unit and referring to the prior studies, an SD of 3.1 was used as the between-cow variability. The projected sample size was nine cows per treatment. The resulted sample size was doubled and the experiment was conducted assigning 18 cows per treatment and it was assumed that the digestibility measured with 18 cows was adequate to detect treatment differences reliably. In each farm, 54 multiparous Holstein lactating dairy cows  were randomly assigned to three pens in one enclosed barn having individual feeding compartments to have 18 cows in each pen. Three dietary treatments were assigned randomly across pens on each farm. The length of each feeding trial was 28 d and during the experiment, cows were fed twice per day at 0600 and 1600 h for 5% refusals. Milking was done twice daily at 0500 and 1400 h and individual daily milk yield (kg/d) of cows was recorded electronically. Daily individual DMI was determined by measuring TMR provided (DM basis) and subtracting the orts remaining (DM basis). Cow weights were recorded on d 1 and 28 in each trial, and initial body weight (BW) of cows was averaged 572 kg (±23 kg). During the feeding trial, TMR samples (500 g each) and orts (200 g) were collected daily (frozen at −20 °C) and composited weekly. Six fecal grab samples (~200 g of feces each) from each cow were collected twice daily to cover 0400, 0800, 1200, 1600, 2000, and 2400 h time points over final 3 days as previously described ,15 and cComposited samples were dried at 60 °C for 48 h in a forced-air oven and were ground through a 1–mm screen . Ground samples were analyzed in duplicate for DM by oven-drying at 105 °C for three hours , ash , CP , ether extract   and NDF Indigestible NDF (iNDF) of diets, feces and orts was obtained as previously described  by weighp ≤ 0.05 significance level). Five sample sizes per each treatment group; N18 , N15 (data from 15 cows), N12 (data from 12 cows) N9 (data from nine cows) and N6 (data from six cows) were simulated by simple random sampling of data from18 cows. Then, 1000 replicates for the five sample sizes were simulated by the bootstrapping method using PROC SURVEYSELECT procedure of SAS with strata specified as each treatment group [p ≤ 0.05 and a trend when p > 0.05 to p ≤ 0.10, respectively. Initial assignment of 18 cows per treatment was used as the reference and based on the initial statistical power calculation, it was assumed as adequate to reliably detect treatment differences  when using data from all 18 cows. Mean body weight and DMI of three diets did not changed with reducing sample size from 18 to 6 per treatment (p > 0.05).The effect of the experimental diet on body weight of cows and DMI are reported in p = 0.01) while STRD increased linearly (p = 0.04). The digestibility of NDF was not affected by diet (p = 0.07) when using data from all 18 cows. Mean DMD, NDFD, and STRD were not different when reducing sample size from 18 to 12. However, further reduction of sample size from 12 to 9 and 6 altered mean digestibility of three digestion variables (p ≤ 0.05).The effect of the experimental diet on digestibility of DM, NDF and starch in lactating dairy cows are reported in p ≤ 0.05). Production of milk fat and milk protein were not different among dietary treatments (p > 0.05). The mean actual daily milk yield was different among five sample size groups (p ≤ 0.05) and according to the results, the mean actual milk yield in N18, N15 and N12 were not different in all three dietary treatment groups. However, reducing the sample size from 12 to 9 and further, milk yields changed. 4% FCM yield and the milk composition were not different among five sample size groups (p > 0.05). The effect of the experimental diet on milk yield, contents and production of milk protein and fat in lactating dairy cows are reported in p ≤ 0.05). Observed SD values for DMD, NDFD, and STRD ranged from 3.8 to 6.3, 5.1 to 9.3 and 3.3 to 5.6 respectively among different sample size groups  in nutrient digestibility between and within-farms are presented in The accuracy of five sample sizes was tested using 1000 bootstrap replicates, and according to our results, the amount of uncertainty associated with an estimate was less regardless of random sampling events as mean digestion variables of original five sample sizes fell within 95% confidence interval estimates of bootstrap replicates. Bootstrapping is a statistical procedure that re-samples a single original dataset to create many simulated samples ,21. WhenIn lactating dairy cows, DMI is driven by milk production, yet physical fill of the reticulorumen and metabolic-feedback factors could limit the intake ,23. Dry The decrease in DMD with increasing NDF:starch ratio observed in the current experiment was mostly caused by the replacement of highly digestible ground corn and cornmeal with low digestible soy hulls. However, NDFD was not different across experimental diets. This could be mostly caused by associative effects of NDF digestibility of corn silage and concentrates on three diets. Apart from that, the diet includes compensatory NDF digestion in the hindgut, and lack of differences among diets in carbohydrate fractions could be other contributing factors for the observed similar DMI. Comparable to our results, one study reported that decreasing NDF:starch ratios from 1.24 to 0.74 did not affect the NDF digestibility of corn silage-based diets when confounding factors are reduced . In anotAlthough actual milk yield and 4% FCM yield were not affected by dietary treatments in the current experiment, we observed an increase in milk fat content from D1 to D3. Increments in NDF concentrations of diets tend to surge the proportion of acetate in the rumen which serves as the main precursor for milk fat synthesis ,32. ThusDecrease of milk protein content with increasing NDF:starch ratio in the current experiment could be partially described within the experimental conditions by low dietary rumen fermentable carbohydrate content resulting from the replacement of highly digestible ground corn and cornmeal with low digestible soy hulls. This is well documented in many previous studies that milk protein content could increase as grain replaced forage or fibrous byproducts to result in low NDF:starch ratio ,34,35. IEstimated statistical power at five sample sizes contradicted initial statistical power projections at the research planning stage and was mostly caused by observed high between-cow variability. The statistical power calculation is the most scientifically favored method used in animal studies in determining an appropriate sample size before the study, which ensures high reliability of the conclusions ,37. HoweDiet was the greatest contributor to the observed total variability farm-to-farm as imposed by treatments. As a general pattern, the effect of between-cow variability was less for highly digestible nutrients  and more for lower digestible nutrients among farms . The farm was a substantial source of variation for the observed between-cow variability for all three nutrient digestion variables and this indicates that the effect of between-cow variation in nutrient digestibility of corn silage-based diets is farm specific. Differences among between-cow variability of nutrient digestion farm to farm could cause by genetic differences among cow groups, farm-specific housing and management factors related to cow nutrition such as accessibility to water and water quality, feeding management and likely several other unidentified factors ,40. ThusObserved between-cow variability in nutrient digestibility within-farm could be due to differences in DM intake, alteration of digestion and mean retention time in the rumen, changes in the rumen microbial population, anatomical differences, sorting behavior during intake and likely several other unidentified factors ,42. Thou2 = 0.30), and according to the author, the passage rate of NDF from the rumen increased with increasing NDF intake due to the substantial variation in NDFD between cows, irrespective of intake [The highest between-cow variability observed for NDFD could be partially explained by diet characteristics and changes of passage rate of NDF from the rumen. A previous study reported a slight negative relationship between NDF intake and NDFD , sampling frequency and variations in the iNDF method could contribute to the over-estimation of the between-cow variability. Although the total collection of feces is the most accurate procedure in quantifying nutrient digestibility , we adapSubstantial between-cow variation in nutrient digestibility exists for diets with different NDF:starch ratio when fed to groups of cows in different farms. However, for a specific dietary treatment arrangement within-farm, between-cow variation in digestibility is not necessarily partitioned the same for different diets and dietary nutrients. Between-cow variation within-farm was highest for NDFD, followed by DMD and STRD, respectively. Thus, evaluating a given dietary treatment effect on NDFD is a primary objective, the sample size will need to be greater  than if the digestibility of other components is the primary objective when fed lactating dairy cows in groups with randomized block designs under the conditions of this experiment.


The development of scleritis in the setting of autoimmune conditions has been well documented. Prior series have assessed the relationship between systemic autoimmune disorders and scleritis only in patients referred for rheumatologic or ocular inflammation. This can lead to a referral bias. We reviewed all charts within the electronic medical record (EMR) of a health system for patients with systemic autoimmune and scleritis diagnoses to determine the prevalence of both and which disorders had the highest relative risk of developing scleritis.The EMR was searched for scleritis and systemic inflammatory diagnoses in the past medical history and diagnosis tabs, and for associated disease specific laboratory values. The intersection of scleritis and systemic inflammatory conditions was assessed through searching both SNOMED Clinical Terminology and ICD-10 codes for diagnoses. The prevalence of each autoimmune disorder, scleritis prevalence, the percentage of patients with an autoimmune condition having scleritis, the percentage of patients with scleritis having an autoimmune condition; the relative risk (RR) of scleritis patients having a specific autoimmune disorder were calculated.n = 26,680; prevalence 0.45%); rheumatoid arthritis (RA). Conversely, 2702 patients were identified with scleritis , of which 31.4% had an associated autoimmune condition. Patients with RA represented the highest percentage of patients with an autoimmune condition having scleritis. Granulomatosis with polyangiitis (GPA) represented the highest the percentage of patients with scleritis having an autoimmune condition. Sjogrens was the third most common condition associated with scleritis- making up 4.5% of cases. An association with juvenile idiopathic arthritis (JIA) was seen in 0.3% of patients.A total of 5.9 million charts were searched with autoimmune conditions identified in 148,993 patients. The most common autoimmune conditions overall were HLA-B27-associated diseases (While this is the largest retrospective review examining the association between autoimmune disease and scleritis, the findings are similar to prior studies with nearly a third of scleritis patients having an underlying autoimmune diagnosis. Limitations of the study included accurate chart coding; having laboratory results within the searchable EMR. Future research is needed to delineate associations of systemic disease with the anatomic location of scleritis using EMR. The development of scleritis in the setting of autoimmune conditions has been well documented –10. An uMany of these series have assessed the relationship between systemic autoimmune disorders and scleritis in patients referred for rheumatologic evaluation and to rule out ocular inflammation –16. LimiThus, we reviewed all of the electronic medical records within the Johns Hopkins Health System to extract patients with both systemic, autoimmune, and scleritis diagnoses. From these, we determined the prevalence of both the autoimmune disorders in addition to scleritis. We further examined which of these systemic disorders had the highest relative risk of developing scleritis.The electronic medical record (EMR) at Johns Hopkins Medicine Health System was queried using the SlicerDicer application which allowed a search of de-identified medical records from 1/1/2013, when the hospital switched to an EMR, through 8/16/2018. Medical records were searched for scleritis; systemic inflammatory conditions in the past medical history or diagnosis tabs, and associated specific laboratory values;  and the intersection of scleritis and systemic inflammatory conditions. Diagnoses were searched by both SNOMED Clinical Terminology  and ICD-10 codes. The query terms are shown in Table p-values. E-values were also calculated for the RR and lower limit of the 95% CI to estimate how strong a confounder would have to be to reduce the association to chance [After totaling the number of individual charts for each condition, the intersection of each systemic autoimmune condition and scleritis was then queried and tallied. The statistics calculated from the data included: the prevalence of systemic autoimmune conditions, the prevalence of scleritis, the percentage of patients with an autoimmune condition having scleritis, the percentage of patients with scleritis having an autoimmune condition, the relative risk (RR) of patients with scleritis also having a specific autoimmune condition with an associated 95% confidence interval (95% CI), and chi-square o chance .n = 47,712, prevalence 0.81%), HLA-B27-associated diseases  and RA . The least common was Behcet disease (n = 317), PAN (n = 365), and GPA (n = 859). Conversely, 2702 patients were identified with scleritis , of which 30.8% had an associated autoimmune condition. The raw data for all diagnoses, number of scleritis cases, percentage of total scleritis associated with each systemic condition, and prevalence of scleritis within each condition are listed in Table A total of 5.9 million charts were searched, and of these, 148,993 patients were identified with an autoimmune condition. The most common autoimmune condition was gout (The most frequent cause of scleritis was RA making up 6.8%, followed by HLA-B27 with 5.7% of cases, and Sjogren / Sicca with 4.5% of cases. The rarest causes were JIA and Behcet  disease at 0.3% of cases followed by PAN at 0.4%.p-values < 0.1 × 10− 10. Full results for associations with 95% confidence intervals, p-values, e-values, and e-values for the 95% CI lower limit are available in Table Many systemic conditions had high association with scleritis. GPA was highest, followed by PAN, Behcet Disease, Sjogren, and RA. RR ranged from 117–20 with all To our knowledge, this is the largest retrospective review examining the association between rheumatologic disease and scleritis. The previous literature is comprised of retrospective analyses of patients seen only within rheumatology or ocular immunology clinics, potentially leading to a referral bias –16. In cThe most common systemic condition associated with the development of scleritis in previous studies has been RA –13, 15. The most interesting findings of the study were the association of Sjogren syndrome and JIA with scleritis in 4.5 and 0.3% of patients, respectively. There have been no prior reported cases of scleritis and JIA, only one case report of peripheral ulcerative keratitis . SimilarThis leads to an important point in regards to Sjogren syndrome, which is labelled as primary if another rheumatologic disease is not diagnosed through a laboratory work-up and physical examination. Sjogren syndrome can also be secondary if another unifying diagnosis is made and also included as a diagnosis code, which highlights the inability to discern the difference using ICD-10 and SNOMED terms. This may partially explain the strong association with Sjogren syndrome with scleritis, as previously only case reports and the minority of patients in longitudinal series described the occurrence of scleritis is patients with Sjogren syndrome –25.Given potential pitfalls and limitations with the use large data analyses, future research is needed to delineate associations of systemic disease with the anatomic location and necrotizing subset of scleritis using electronic medical records. However, the results of this study can assist clinicians in broadening their differential diagnoses and work-up during the evaluation of patients with autoimmune conditions and scleritis.


Obesity is considered a top public health concern, and its prevalence is growing every day. Thus, interventions to address this problem should be encouraged and further studied. In this regard, the aim of this review was to summarize the evidence of martial arts interventions to evaluate their effectiveness on the anthropometric and body composition parameters of overweight and obese subjects.A systematic literature search was conducted on January 26, 2020 using the PubMed, Medline, Lilacs, Cochrane, and Scielo databases. Reference lists of eligible articles and relevant reviews have also been examined. All randomized clinical trials on martial arts that evaluated the anthropometric and body composition parameters of overweight and obese subjects were included, and a narrative synthesis of eligible studies was conducted in accordance with PRISMA guidelines. The Downs & Black checklist was used to assess the quality of the studies. This review was registered in the International Prospective Register of Systematic Reviews (PROSPERO) (identifier CRD42018086116).2; 95% CI: − 2.72, 0.05), waist circumference  and percentage of body fat .A total of 82 articles were identified from the initial search strategy. A further 2 articles were identified from the review of relevant bibliographies. Six studies encompassing 258 participants who were overweight or obese were included. Four studies reported Tai Chi practice, one study reported Kung Fu exercise, and another study reported martial arts exercise. The examined meta-analyses did not reveal significant benefits from martial arts practice over control groups after the experiment period for body mass index (− 1.34 kg/mThe scarcity, heterogeneity, short intervention time, small sample size, and significant methodological limitations of the available studies do not allow to conclude whether martial arts are effective in the anthropometric and body composition parameters of overweight and obese individuals. This study highlights the need for more research to assess the benefits of martial arts for overweight and obese individuals. Obesity is a chronic disorder of multifactorial origin, defined as an accumulation of abnormal or excessive fat that can be detrimental to health . CurrentLifestyle interventions that provide a 5–10% reduction in body weight are recommended as a strategy to combat obesity , 4. HoweInterventions for treating overweight and obese people include nutritional education and changes in lifestyle, which require increasing levels of physical activity and the reduction of sedentary behaviors . The incAn organized activity that emerges as an alternative form of physical activity and that is gaining many supporters is the practice of martial arts –21. MartDespite the studies reporting and analyzing the benefits of practicing martial arts in the general population, scientific evidence regarding their use for the treatment of overweight and obese subjects is limited. Therefore, before recommending martial arts as a treatment for weight loss, it is important to analyze the scientific evidence for potential benefits and risks. For that reason, the purpose of this systematic review was to evaluate the effectiveness of martial arts exercise on the anthropometric and body composition parameters of overweight and obese subjects.The PRISMA Statement   was usedThe inclusion criteria were the following: (1) randomized controlled trials study design; (2) published in English, Spanish or Portuguese; (3) martial arts exercise, isolated or combined with any other practices; (4) overweight or obese participants ; (5) use of a quantitative method to determine at least one of the relevant measurements of anthropometric and body composition parameters .To ensure the inclusion of relevant literature, the electronic search was conducted in different databases, namely PubMed, Medline, Lilacs, Cochrane, and Scielo in January 26, 2020. The adopted search strategy included a combination of the following Medical Subject Headings (MeSH) terms: obesity AND martial arts. Keyword iterations related to these terms were also used. Example of the search strategy in PubMed: Obesity [all fields]; Obes* [all fields]; Overweight [all fields]; #1 OR #2 OR #3; Martial arts [all fields]; Martial Fitness [all fields]; Martial exerc* [all fields]; #5 OR #6 OR #7; #4 AND #8 . Each researcher selected the studies that met the inclusion criteria. Full texts were read by 2 authors  and the final list of eligible studies was compiled. If discrepancies between the 2 researchers occurred those items were discussed with a third researcher (DJT) to achieve a consensus. After selecting the articles, the following data were extracted: author, date of publication, language, sample size, demographic characteristics , BMI, body composition, intervention and control, and outcomes. The researchers were not blinded to the journal titles and paper authors for the data extraction process.The methodological quality of the studies was evaluated independently by two researchers (FS and FNL), using the Downs & Black checklist . This asThe systematic review data were analyzed and presented employing a narrative synthesis approach. We examined the year of publication, country of origin, study design, sample characteristics, types of martial arts selected for intervention, outcome measures and treatments applied to the control groups. The overall effect sizes for each outcome were determined using meta-analytic approaches if at least 2 studies assessed the specific outcome. The Review Manager 5 software  was used, and random effects models were applied. Meta-analyses were conducted to examine the effects of martial arts compared with control groups. Mean differences (MD) with 95% confidence intervals (CI) were reported. Attempts were made to obtain missing data from the studies’ authors by email.2 statistics, and the magnitude of heterogeneity was categorized as I2 = 0 to 24%: low; I2 = 25 to 49%: moderate; I2 = 50 to 74%: substantial; and I2 = 75 to 100%: considerable heterogeneity [2 test was used to assess the statistical significance of heterogeneity between trials. In the face of the low power of this test in small samples, a P value ≤0.10 was regarded to indicate significant heterogeneity [Statistical heterogeneity between the studies was determined using Iogeneity , 28; andogeneity .The electronic search initially retrieved 82 titles. Two additional articles were found in the reference section of the review articles. After duplicate articles were removed, 66 articles were screened by reading the titles and abstracts. Of them, 52 articles were excluded because they did not meet the inclusion criteria. The remaining 14 articles were selected for further review. After full reading, 8 studies were excluded . The study selection process is shown in the PRISMA  flow dian = 6) [n = 187), with a mean age of 46.5 years (SD = 18.66). BMI was taken as a criterion in all studies for identifying the overweight or obese sample 100% (n = 6), but different criteria were adopted in various studies to define overweight and obesity. Most studies (66.66%) defined a BMI ≥ 30 (n = 4) as a criterion for inclusion, one study (16.66%) defined a BMI ≥ 25, and one study (16.66%) defined a BMI ≥ 85th percentile.Six randomized clinical trials (n = 6) –35 were n = 6) , 33, 35,n = 6) , one in n = 6) , and onen = 6) . The anaRegarding the type of intervention, four studies (66.66%) reported Tai Chi practice as an intervention technique , 34, 35,n = 4). The Yang style 24-posture Form was the most widely studied 75% (n = 3) [Tai Chi was the most common martial arts training applied as intervention in the treatment group 66.66% ( (n = 3) , 34, 35, (n = 3)  reported (n = 3) , 34, 35  (n = 3)  adopted Different interventions were performed in the control groups applying Tai Chi as the main intervention in the treatment group , 34, 35.Two studies 33.33%) , 33 deve.33% , 33Three studies have not reported the participation rate in their interventions , 32, 34.Two studies , 35, whin = 4), and 33.33% were rated low quality (n = 2); none was rated high quality. Assessment of study quality in this systematic review revealed the following negative factors: no identification of the main confounding factors, lack of treatment control and details of adverse events, poor analysis of the participants lost in the follow-up, absence of random sampling to represent the entire population from where the participants were recruited, no blinding for those responsible for assessing the outcomes, lack of adequate adjustment for the confounding variables in data analysis from which the findings were drawn, and inadequate sample size.A quality index was assigned to each of the studies included in this systematic review. The score ranged from 11 to 21, with a mean of 17.66 (SD = 3.98) , as showEffectiveness assessment of martial arts practice in overweight and obese subjects are presented as follows.2; SD = 4.2) and control groups  after the intervention period, with a lower BMI in the intervention group. However, four studies (66.66%) [All studies of this systematic review assessed BMI (100%), but only Chen and colleagues (16.66%)  reported(66.66%) , 35 repo2 = 0%), with p = 0.06 and low level of heterogeneity.Five studies (83.33%) –34 in thp < 0.005). Nonetheless, two studies (33.33%) [Three studies (50%) assessed the percentage of body fat , 34, 35 2 = 82%), with p = 0.76 and high level of heterogeneity.Three studies (50%) , 34, 35 Three studies (50%) assessed waist circumference , 31, 35 2 = 0%), with p = 0.19 and low level of heterogeneity.Three studies (50%) , 31, 35 This systematic review evaluated the efficacy of martial arts training on the anthropometric and body composition parameters of overweight and obese subjects. It can be inferred that martial arts are a promising form of intervention for the treatment of overweight and obese subjects, since the meta-analysis showed a tendency to decrease BMI in those groups that received martial arts interventions Fig. . HoweverThe results observed in this systematic review regarding the anthropometric and body composition parameters were incongruent, since some studies verified significant differences after the intervention period, whereas others did not. One factor that may have influenced the results was related to the age of the participants, since most of the studies –35 were A recent systematic review has investigated the benefits provided by the practice of hard martial arts in adults, and found they were safe and could provide several benefits, even for the elderly population . HoweverAnother study by Chyu and colleagues  adopted The studies showing positive outcomes regarding changes in body composition used Tai Chi intervention followed by nutrition education and dietary intervention , 35. ThiOne of the challenges encountered in this review was the fact that all the studies adopted different methods, with different frequency and duration of training sessions, treatment lengths of time, and additional treatment interventions in the control groups, making it difficult to compare the studies. Another difficulty in pursuing the objectives of this review was the fact that most of the studies focused on Tai Chi as the martial art intervention, which is a low-intensity martial art.This systematic review did not identify any major adverse events in any of the studies that composed it. However, 33.33% of the studies did not mention whether there were adverse events in their interventions, which could lead to misinterpretation of the safety of the interventions.The results of the quality assessment of the studies that composed this review show that most studies obtained a medium-quality rating and none of them obtained a high score, which indicates more rigorous research studies are needed to determine the efficacy of the interventions Table . Two of Given that this was the first systematic review to examine the effects of martial arts interventions on overweight and obese subjects, all randomized clinical trials published up to January 2020 that met the inclusion criteria were encompassed in the review, regardless of their quality assessment.Despite the limitations in this study, it should be highlighted that this was the first systematic review to assess martial arts exercise interventions for overweight and obese subjects.This systematic review presented important outcomes from the studies that composed it. However, although the interventions have been relatively safe, it is important to conduct further research involving other types of martial arts, with more rigorous study designs, involving nutritional intervention with control of energy intake, assessment and control of daily activities, detailed description of the interventions and adverse events, larger sample sizes, and different research populations. These measures would allow us to obtain more conclusive results on the effects of martial arts as a treatment method for overweight and obese subjects. They would also help obtain results that can be generalized with confidence.The scarcity, heterogeneity and significant methodological limitations of the available studies do not allow to conclude whether martial arts are effective in the anthropometric and body composition parameters of overweight and obese individuals. Martial arts can be an important tool for the treatment of overweight and obese people, as the results of this systematic review indicate that martial arts are probably safe and may offer benefits related to anthropometric and body composition parameters. However, the results are limited and should be used with caution. Because of this, further research is needed before we can generalize the findings regarding the efficacy of martial arts for maintaining healthy anthropometric parameters among overweight and obese subjects.Additional file 1. Search strategy.Additional file 2. Checklist for measuring study quality (Downs & Black).


The usefulness of pulse oximetry for the management of obstructive sleep apnea is controversial. The aim of this study was to assess the accuracy for indication of Continuous Positive Airway Pressure (CPAP) treatment in patients with suspected obstructive sleep apnea (OSA) based on clinical and oximetry data as compared to polysomnography (PSG). This multicenter observational study involved seven sleep laboratories. Patients with suspicion of OSA who completed a standardized sleep questionnaire and a diagnostic PSG were enrolled. Eight observers logged on to a website independently and blindly. Seven observers only accessed the clinical data, curve and pulse oximetry results (Os-SO2-test method), while the eighth observer had full access to all indicators of PSG (O-PSG-reference method). Once observers assessed the information available on the website, they had to choose between three CPAP treatment options (yes/no/do not know) based on their knowledge and criteria.p 0.028); Sp 93.8-100% versus 68.5-96.6%, p 0.004; PLR > 10 versus 2.9-17, p<0.01). 411 subjects (228 men), median age 54 years, were available for evaluation. Os-SO2 had lower sensitivity (S), greater specificity (Sp) and positive likelihood ratio (PLR) to prescribe CPAP in patients more symptomatic  than those with fewer symptoms (ESS < 11 without comorbidities) (S 45-75% versus 45-91%,  Due to its low false positive rate, a strategy based on pulse oximetry and clinical data was a consistent tool to indicate CPAP treatment in most symptomatic patients with a suspicion of OSA. Lázaro Langer), Mendoza (Instituto Médico Insares), Salta  and Paraná (Centro Privado de Medicina Respiratoria de Paraná). It was approved by the Ethics Committee of Hospital Alemán.On a database of 761 patients (304 women) studied in the Hospital Aleman’s sleep laboratory between 2013 and 2015, an intentional sampling was carried out in order to have a similar proportion of men and women with a hypopnea apnea index (AHI) less than 5 and with mild, moderate and severe OSA. Patients over 18 years of age who had undergone PSG and filled out a standardized sleep questionnaire (SQ) were preselected. Exclusion criteria were: patients with a total sleep time of less than 180 minutes, PSG with CPAP titration, oximetry with artifacts by disconnections or finger probe displacement more than 5% of total recording time, subjects with suspected narcolepsy or central sleep apnea by PSG2 readings. Software on PSG performed automatic SO2 analysis and excluded areas with artifacts (artifact time). Then, the program calculated the oxygen desaturation index (ODI) ≥ 3%  using the valid total recording time . ODI3 was selected to assess pulse oximetry as it has shown the best intraclass correlation coefficient with AHI and therefore could be used as surrogate for AHIThe patients selected for this study had a PSG with a computerized polysomnographic system  which included: electroencephalogram , electrooculogram, chin electromyogram, leg electromyogram, electrocardiogram, airflow by nasal pressure and an oral thermistor, thoracic and abdominal movements (piezoelectric sensors), snoring, SO2 and body position. PSG interpretation was performed according to published criteria,Upon arrival to the sleep lab, the patients completed a standardized sleep questionnaire (SQ) that included the Spanish versions of the Epworth Sleepiness Scale (ESS)21Comorbidities such as hypertension, coronary disease, ischemic or hemorrhagic stroke or diabetes type II, were considered when patients reported them, or if they received any medication for their condition. Restless leg syndrome was assumed when patients complained of unpleasant sensations in the lower limbs before sleeping and reported behaviors to alleviate them. Tiredness was regarded as a symptom when it was described on a daily basis. Insomnia was defined both as difficulty falling asleep or waking up while sleeping and delay in returning to sleep more than 5 times a month.The database with all the information necessary for decision making was available on a website specifically designed for this research. Researchers could access it through username and password to see clinical data, questionnaires, oximetry or PSG data and treatment decision options.2), while the eighth observer, who also had full access to all indicators of PSG, was the reference method (O-PSG). Once observers assessed the information available on the website, they had to choose between three CPAP treatment options . The term “do not know” could be selected when the observers had doubts in the prescription of CPAP or could not make the decision with the information available. Observers 1 to 7 could indicate CPAP treatment if the patients met clinical criteria and their oximetry was compatible with OSA. O-PSG and Os-SO2 supported CPAP treatment decision on their experience and current published guidelines23,Three to six months before starting the study, all researchers received written instructions to access and use the website tools so that they could train with twenty clinical cases to understand work dynamics. To help decide treatment with CPAP, all observers used a summary sleep questionnaire (SSQ). Eight physicians with at least 10 year experience in the management of patients with sleep-related breathing disorders independently logged onto a website and were blinded to any patient identification data or indication of CPAP. Seven observers (one per center) who could only access the clinical data (SSQ), curve and pulse oximetry results were collectively referred to as the test method  for the indication of CPAP was evaluated by calculating the sensitivity (S), specificity (Sp), positive and negative likelihood ratio  in each observer for both the total group and the more or less symptomatic patients. The more symptomatic patients were defined as the presence of Epworth greater than 10 and / or at least one comorbidity, while the less symptomatic subjects were those with an Epworth less than 11 and without comorbiditiesThe accuracy of the Os-SOMultiple logistic regression analysis was used to assess the association between CPAP indication most frequently made by Os-SO2 in each patient  and age, gender , BMI, sleepiness , comorbidities , insomnia , restless leg syndrome  and ODI3 ≥ 15  (independent variables). ODI3 ≥ 15 was chosen because this cut-off point has shown good performance to diagnose moderate to severe OSAp 0.8). ESS > 10 was present in 38.2% of patients and tiredness was reported by 54%.Of 761 potentially eligible patients, 350 were excluded for different reasons see . Thus, tO-PSG indicated CPAP to 225 subjects (54.7%) of which 85% presented moderate to severe OSA  and 80% had either sleepiness or at least one comorbidity.2 are shown in 2 had average S, Sp and PLR of 61.7%, 93.4% and 19.3, respectively. Os-SO2 had lower S, greater Sp and PLR for CPAP indication in more symptomatic patients than those with fewer symptoms .S, Sp and PLR / NLR for Os-SO2, reported the highest number of false positive (FP) and negative (FN) cases. Two Os-SOp<0.0001). Also, being male , an ESS > 10  and some comorbidities  were independently associated with CPAP indication.2, which had 80 false-negative subjects and no false positive case, and the average costs in the local market. Based on these data, we could have saved US$ 24200  and experience of physicians to indicate CPAP treatment in more symptomatic patients. For this, we considered the decision of treatment with CPAP most frequently made by Os-SOS$ 24200 .2, both in the total group (45 - 79%) and subgroups of more (45 - 75%) or less symptomatic (45 - 87%) patients. This interobserver heterogeneity possibly reflects the complexity of the CPAP treatment decision-making procedure, which involves a wide range of variables, such as symptoms (sleepiness or tiredness being the most important), associated comorbidities and physician´s experience with OSA patients and the interpretation of pulse oximetry. In our patient population, the pretest probability, (i.e. CPAP recommendation by PSG) was 55% in the total group, 65% in the more symptomatic patients and 34% in those with fewer symptoms. Depending on the PLR, the posttest probability of obtaining a true positive diagnosis ranged from 60 to 90% in oligosymptomatic subjects, 80 to 97% in the total group and 95 to 99% in the more symptomatic patients2 observers, Sp was similar  and PLR was greater than 10 in more symptomatic patients. The low proportion of false positive subjects makes this approach a consistent tool. In our cohort, observer 4 showed the highest number of FP cases. However, when analyzing these 34 cases, 30 patients could have had CPAP indication according to the published guidelines2 did not). In this way, patients suspected of obstructive sleep apnea but with inconclusive oximetry would require a second diagnostic sleep study with the consequent potential increase in costs and treatment delay. Despite these disadvantages, if this strategy had been adopted only in more symptomatic patients, the health costs could have been reduced by almost 20% compared to the traditional polysomnography-based approach , the number of observers who participated in both studies (seven versus one) and the fact that observers´ decisions were based exclusively on CPAP treatment guidelines without resorting to their experience.We recently publishedOur results are in line with previous clinical validation studies of oximetry. Mulgrew et al.This study has several strengths. It is a multicenter study of a large sample of patients and a similar number of men and women. Also, the proportion of subjects without or with mild, moderate or severe OSA was similar. All patients had a standardized sleep questionnaire and a PSG. Finally, the observers who participated were blind and the therapeutic decision to or not to treat with CPAP was based on each observer´s knowledge and experience so as to simulate a real-life scenario. However, this clinical research presents some limitations. First, its retrospective design and selection bias limits its generalization. Second, this simple strategy requires training in the management of patients with sleep apnea and analysis of oximetry, therefore it cannot be extrapolated to general practitioners or primary care physicians.In conclusion, a tool based on clinical data and pulse oximetry allowed to indicate CPAP treatment reliably in most symptomatic patients with suspicion of OSA. This approach would be useful in the initial management of OSA when access to specialized sleep medicine centers is limited for different reasons. Future studies are necessary to confirm these results in a real life scenario.


Siphonosoma cumanense is economic important species in the fishery of southeast China. However, the current classification and the phylogeny of genus Siphonosoma had not been verified yet. Here, we report the complete mitochondrial genome sequence of S. Siphonosoma. The mitogenome has 15,917 base pairs and made up of total of 38 genes , and a putative control region. This study was the first available complete mitogenomes of Siphonosoma and will provide useful genetic information for future phylogenetic and taxonomic classification of Sipuncula. Siphonosoma cumanense is an important fisheries resource for its valuable nutrition and valuable ingredients of chines traditional medicines. However, the current classification and the phylogeny of genus Siphonosoma has not yet been determined. Mitochondrial genome is useful molecular techniques for phylogenetic relationships and taxonomy identification, especially when there is small number of unambiguous taxonomic characters available for taxonomy determination. Here, we report the first complete mitochondrial genome sequence of Siphonosoma, which will provide a better insight into phylogenetic assessment and taxonomic classification of Sipuncula.Sipuncula (known as peanut worms) is an ancient group of exclusively marine worms which contain about 150 exclusively species with a global distribution , and the whole body specimens (#GG0241) were deposited at Marine biological Herbarium, Guangxi Institute of Oceanology, Beihai, China. The total genomic DNA was extracted from the muscle of the specimens using an SQ Tissue DNA Kit  following the manufacturer’s protocol. DNA libraries (350 bp insert) were constructed with the TruSeq NanoTM kit  and were sequenced (2 × 150 bp paired-end) using HiSeq platform at Novogene Company, China. Mitogenome assembly was performed by MITObim  was chosen as the initial reference sequence for MITObim assembly. Gene annotation was performed by MITOS (http://mitos2.bioinf.uni-leipzig.de/).The tissue samples of S. Siphonosoma from Beibu Bay was found to be 15,917 bp in length (GenBank accession number: MN813483), including the usual set of gene for mitogenome except for extra threonine tRNA which consisting of 13 protein-coding, 23 tRNA and 2 rRNA genes, and a putative control region. The overall base composition of the mitogenome was estimated to be A 27.5%, T 33.2%, C 25.4% and G 13.9%, with a high A + T content of 60.8%, which is slightly higher than Sipunculus nudus (57.5%)  (Zhao et al. S. Siphonosoma was first clustered with genus Sipunculus and then clustered with genus Phascolosoma (Siphonosoma had been revised within the new family Siphonosomatidae (Kawauchi et al. Siphonosoma need further study. Our study suggested closely related taxa between genus Siphonosoma and Sipunculus, which will contribute to further phylogenetic and taxonomic study of sipunculan worms.The complete mitogenome of colosoma , which icolosoma . In the