bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation, characterization and application of theophylline-degrading Aspergillus fungi
Authors: ZHOU Bin-Xing , Ma Cunqiang , Xia Tao , Li Xiaohong , Zheng Chengqin , Wu Tingting , Liu Xiaohui
Published: March 2020

Background: Caffeine, theobromine and theophylline are main purine alkaloid in tea. Theophylline is the downstream metabolite and it remains at a very low level in Camellia sinensis. In our previous study, Aspergillus sydowii could convert caffeine into theophylline in solid-state fermentation of pu-erh tea through N-demethylation. In this study, tea-derived fungi caused theophylline degradation in the solid-state fermentation. The purpose of this study is identify and isolate theophylline-degrading fungi and investigate their application in production of methylxanthines with theophylline as feedstock through microbial conversion. Results:Sseven tea-derived fungi were collected and identified by ITS, β-tubulin and calmodulin gene sequences, Aspergillus ustus, Aspergillus tamarii, Aspergillus niger and Aspergillus sydowii associated with solid-state fermentation of pu-erh tea have shown ability to degrade theophylline in liquid culture. Particularly, A. ustus and A. tamarii could degrade theophylline highly significantly (p < 0.01). 1,3-Dimethyluric acid, 3-methylxanthine, 3-methyluric acid, xanthine and uric acid were detected consecutively by HPLC in A. ustus and A. tamarii, respectively. The data from absolute quantification analysis suggested that 3-methylxanthine and xanthine were the main degraded metabolites in A. ustus and A. tamarii, respectively. 129.48 ± 5.81 mg/L of 3-methylxanthine and 159.11 ± 10.8 mg/L of xanthine were produced by A. ustus and A. tamarii in 300 mg/L of theophylline liquid medium, respectively. Conclusions: For the first time, we confirmed that isolated Aspergillus ustus, Aspergillus tamarii degrade theophylline through N-demethylation and oxidation. We were able to biologically produce 3-methylxanthine and xanthine efficiently from theophylline through a new microbial synthesis platform with A. ustus and A. tamarii as appropriate starter strains.
Changes in soil microbial communities at Jinsha earthen site are associated with earthen site deterioration
Authors: Li Jing , Zhang Xiaoyue , Xiao Lin , Liu Ke , Li Yue , Zhang Ziwei , Chen Qiang , Ao Xiaolin , Liao Decong , Gu Yunfu , Ma Menggen , Yu Xiumei , Xiang Quanju , Chen Ji , Zhang Xiaoping , Yang Tao , Penttinen Petri , Zhao Ke
Published: March 2020

Background: Earthen sites are immobile cultural relics and an important part of cultural heritage with historical, artistic and scientific values. The deterioration of features in earthen sites result in permanent loss of cultural information, causing immeasurable damage to the study of history and culture. Most research on the deterioration of earthen sites has concentrated on the physicochemical factors, and information on microbial communities in earthen sites and their relationship with the earthen site deterioration is scarce. We used high-throughput sequencing to analyze bacterial and fungal communities in soils from earthen walls with different degree of deterioration at Jinsha earthen site to characterize the communities and their correlation with environmental factors, and compare community structures and the relative abundances of individual taxa associated with different degree of deterioration for identifying possible marker taxa.Results: The relative abundances of Proteobacteria and Firmicutes were higher and that of Actinobacteria lower with higher degree of deterioration. At the genus level, the relative abundances of Rubrobacter were highest in all sample groups except in the most deteriorated samples where that of Bacteroides was highest. The relative abundance of the yeast genus Candida was highest in the severely deteriorated sample group. The bacterial phylum Bacteroidetes and genus Bacteroides, and fungal class Saccharomycetes that includes Candida sp. were specific for the most deteriorated samples. For both bacteria and fungi, the differences in community composition were associated with differences in EC, moisture, pH, and the concentrations of NH4+, K+, Mg2+, Ca2+ and SO42-. Conclusion: The microbial communities in soil with different degree of deterioration were distinctly different, and deterioration was accompanied with bigger changes in the bacterial than in the fungal community. In addition, the deteriorated soil contained higher concentrations of soluble salts. Potentially, the accumulation of Bacteroides and Candida plays an important role in the deterioration of earthen features. Further work is needed to conclude whether controlling the growth of the bacteria and fungi with high relative abundances in the deteriorated samples can be applied to alleviate deterioration.
First Isolation and Molecular Characterization of bla CTX-M-121 -producing Escherichia coli O157:H7 Strain Y4-A109 from Cattle in China
Authors: Su Zhanqiang , Author) Panpan Tong(Former Corresponding , Zhang Ling , Zhang Mengmeng , Wang Dong , Ma Kaiqi , Zhang Yi , Liu Yingyu , Xia Lining , Author) Jinxin Xie(New Corresponding
Published: March 2020

Background: To study the antibiotic resistance, the molecular epidemiology of bovine Escherichia coli ( E.coli ) O157:H7, and exploring the intrinsic relationship among different isolates, we have collected 27 bovine E. coli O157:H7 strains in Xinjiang from 2012 to 2017 and evaluated virulence genes, antibiotic resistance, and pulsed-field gel electrophoresis (PFGE) molecular typing. Results: Of all the 27 bovine E. coli O157:H7 strains analyzed, 21 strains contained at least one virulence gene, 19 strains carried eae gene (70.4%) and 8 of them carrying stx1 + stx2 + eae + hly + tccP . Most strains were sensitive to all the antibiotics tested. However, 4 of which were antibiotic-resistant, and 2 of which possessed multi-drug resistance, including one ESBL-producing strain. This is the first report of the bla CTX-M-121 gene in bovine E. coli O157:H7. Moreover, the bla CTX-M-121 gene can be transmitted horizontally through plasmid between strains. The similarity of PFGE spectra of 27 strains was between 65.8% and 100%. Two types of PFGE were obtained through cluster analysis, including clusters Ⅰ and Ⅱ. Conclusions: E. coli O157:H7 may have undergone clonal propagation in cattle farms as well as cross-regional transmission and horizontal transmission in different regions in Xinjiang China.
Characterization of Cystoisospora suis sexual stages in vitro
Authors: Feix Anna Sophia , Cruz-Bustos Teresa , Ruttkowski Bärbel , Joachim Anja
Published: March 2020

Background: The porcine coccidium Cystoisospora suis is characterized by a complex life-cycle during which asexual multiplication is followed by sexual development with two morphologically distinct cell types, the micro- and macrogametes. Genes related to the sexual stages and cell cycle progression were previously identified in related Apicomplexa. Dynein light chain type 1 and male gamete fusion factor HAP2 are restricted to microgametes. Tyrosin-rich proteins and oocyst wall proteins are a part of the oocyst wall. The Rad51/Dmc1-like protein and Nima-related protein kinases are associated with the cell cycle and fertilization process.Here, the sexual stages of C. suis were characterized in vitro morphologically and for temporal expression changes of the mentioned genes to gain insight into this poorly known phase of coccidian development.Methods: Sexual stages of C. suis developing in vitro in porcine intestinal epithelial cells were examined by light and electron microscopy. The transcriptional levels of genes related to merozoite multiplication and sexual development were evaluated by quantitative real-time PCR at different time points of cultivation. Transcription levels were compared for parasites in culture supernatants at 6–9 days of cultivation (doc) and intracellular parasites at 6–15 doc.Results: Sexual stage of C. suis was deteceted during 8–11 doc in vitro. Microgamonts (16.8 ± 0.9 µm) and macrogamonts (16.6 ± 1.1 µm) are very similar in shape and size. Microgametes had a round body (3.5 ± 0.5 µm) and two flagella (11.2 ± 0.5 µm). Macrogametes were spherical with a diameter of 12.1 ± 0.5 µm. Merozoite gene transcription peaked on 10 doc and then declined. Genes related to the sexual stages and cell cycle showed an upregulation with a peak on 13 doc, after which they declined.Conclusions: The present study linked gene expression changes to the detailed morphological description of C. suis sexual development in vitro, including fertilization, meiosis and oocyst formation in this unique model for coccidian parasites. Following this process at the cellular and molecular level will elucidate details on potential bottlenecks of C. suis development (applicable for coccidian parasites in general) which could be exploited as a novel target for control.
Bacteria drug resistance profile affects knee and hip periprosthetic joint infection outcome with debridement, antibiotics and implant retention
Authors: Rudelli Bruno Alves , Giglio Pedro Nogueira , Carvalho Vladimir Cordeiro , Pecora Jose Ricardo , Gurgel Henrique Melo Campos , Gobbi Riccardo Gomes , Vicente Jose Ricardo Negreiros , Lima Ana Lucia Lei Munhoz , Helito Camilo Partezani
Published: March 2020

BACKGROUND: evaluate the effect of bacteria drug resistance profile on the success rates of debridement, antibiotics and implant retention. METHODS: All early acute periprosthetic infections in hip and knee arthroplasties treated with DAIR at our institution over the period from 2011 to 2015 were retrospectively analyzed. The success rate was evaluated according to the type of organism identified in culture: multidrug-sensitive (MSB), methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant Gram-negative bacteria (MRB) and according to other risk factors for treatment failure. The data were analyzed using univariate and multivariate statistics. RESULTS: Fifty-seven patients were analyzed; there were 37 in the multidrug-sensitive bacteria (MSB) group, 11 in the methicillin-resistant Staphylococcus aureus (MRSA) group and 9 in the other multidrug-resistant Gram-negative bacteria (MRB) group. There was a statistically significant difference (p<0.05) in the treatment failure rate among the three groups: 8.3% for the MSB group, 18.2% for the MRSA group and 55.6% for the MRB group (p=0.005). Among the other risk factors for treatment failure, the presence of inflammatory arthritis presented a failure rate of 45.1 (p<0.05). CONCLUSION: DAIR showed a good success rate in cases of early acute infection by multidrug-sensitive bacteria. In the presence of infection by multidrug-resistant bacteria or association with rheumatic diseases the treatment failure rate was higher and other surgical options should be considered in this specific population. The MRSA group showed intermediate results between MSB and MRB and should be carefully evaluated.
Low doses of Bisphenol S affect post-translational modifications of sperm proteins in male mice
Authors: Řimnáčová Hedvika , Štiavnická Miriam , Moravec Jiří , Chemek Marouane , Kolinko Yaroslav , García-Álvarez Olga , Mouton Peter R. , Trejo Azalia Mariel Carranza , Fenclová Tereza , Eretová Nikola , Hošek Petr , Klein Pavel , Králíčková Milena , Petr Jaroslav , Nevoral Jan
Published: March 2020

Background: Bisphenol S (BPS) is increasingly used as a replacement for bisphenol A in the manufacture of products containing polycarbonates and epoxy resins. However, further studies of BPS exposure are needed for the assessment of health risks to humans. In this study we assessed the potential harmfulness of low-dose BPS on reproduction in male mice.Methods: To simulate human exposure under experimental conditions, 8-week-old outbred ICR male mice received 8 weeks of drinking water containing a broad range of BPS doses [0.001, 1.0, or 100 µg/kg body weight (bw)/day, BPS1-3] or vehicle control. Mice were sacrificed and testicular tissue taken for histological analysis and protein identification by nano-liquid chromatography/mass spectrometry (MS) and sperm collected for immunodetection of acetylated lysine and phosphorylated tyrosine followed by protein characterisation using matrix-assisted laser desorption ionisation time-of-flight MS (MALDI-TOF MS).Results: The results indicate that compared to vehicle, 100 µg/kg/day exposure (BPS3) leads to 1) significant histopathology in testicular tissue; and, 2) higher levels of the histone protein γH2AX, a reliable marker of DNA damage. There were fewer mature spermatozoa in the germ layer in the experimental group treated with 1 µg/kg bw (BPS2). Finally, western blot and MALDI-TOF MS studies showed significant alterations in the sperm acetylome and phosphorylome in mice treated with the lowest exposure (0.001 µg/kg/day; BPS1), although the dose is several times lower than what has been published so far. Conclusions: In summary, this range of qualitative and quantitative findings in young male mice raise the possibility that very low doses of BPS may impair mammalian reproduction through epigenetic modifications of sperm proteins.
‘They are inconveniencing us’ - Exploring how gaps in patient education and patient centred approaches interfere with TB treatment adherence: Perspectives from patients and clinicians in the Free State Province, South Africa.
Authors: Moodley Nishila , Saimen Amashnee , Zakhura Noor Mahomed , Motau David , Setswe Geoff , Charalambous Salome , Chetty-Makkan Candice Maylene
Published: March 2020

Background: Tuberculosis (TB) treatment loss to follow up (LTFU) plays an important contributory role to the staggering TB epidemic in South Africa. Reasons for treatment interruption are poorly understood. Treatment interruption appears to be the culmination of poor health literacy of patients and limited health education provided by clinicians. We explored clinician and patient perspectives of the gaps in TB messaging that influence TB treatment LTFU. Methods: We conducted semi-structured in-depth interviews between January and May 2018 with a sample of 15 clinicians managing TB and 7 patients identified as LTFU in public clinics in the Free State Province, South Africa. We used thematic analysis with deductive and inductive approaches. Results: Limited occupational opportunities, fear of disclosure and stigmatization all contributed to treatment LTFU. Patients felt that the TB messaging received was inadequate. Many of the clinicians interviewed felt that improving patient’s TB knowledge would reinforce adherence to treatment and thus focused on sharing information on treatment completion, side effects and infection control. However, the inability of clinicians to establish rapport with patients or to identify social support challenged TB treatment adherence by patients. Clinicians perceived this as patients did not follow their instructions despite what they considered lengthy TB education. Having said this, clinicians concurred that their medical management of TB lacked the psycho-social dimension to treat a social disease of this magnitude. Conclusions: Limited occupational opportunities, fear of disclosure and stigmatization all contributed to treatment LTFU. Clinicians concurred that poor patient understanding of TB and that biomedical management lacking a psycho-social dimension further exacerbated the poor treatment outcome. TB remains a social disease, the successful management of which hinges on patient-centred care. Keywords: tuberculosis TB, adherence, messaging, loss to follow up, knowledge, health literacy, patient-centered care
Protective effect of hydrogen gas inhalation on radiation-induced bone marrow damage in cancer patients: a retrospective observational study
Authors: Hirano Shin-ichi , Aoki Yukimasa , Kurokawa Ryosuke , Li Xiao-Kang , Ichimaru Naotsugu , Takahara Shiro , Takefuji Yoshiyasu
Published: March 2020

Background: Although intensity-modulated radiation therapy (IMRT) has been developed as an alternative to conventional radiotherapy, but reducing bone marrow damage is limited in the patients with multiple tumor lesions and large irradiation volume. Thus, novel technology is needed to mitigate further IMRT-induced bone marrow damage. Molecular hydrogen (H2) was recently reported as a preventive and therapeutic antioxidant that selectively scavenges hydroxyl radical (·OH) and peroxynitrite (ONOO-). This observational study aims to examine whether H2 gas treatment improves IMRT-induced bone marrow damage in cancer patients. Methods: The patients with end stage of cancer were received IMRT once per day for 1 to 4 weeks except Saturday and Sunday. After each course of IMRT, the patients of control group (n = 7) were housed in health care chamber (HCC, mild hyperbaric oxygen chamber) for 30 minutes, and the patients of H2 group (n = 16) were also housed in HCC and received 5% H2 gas for 30 minutes once per day. Radiation-induced bone marrow damage was evaluated by hematological examination of peripheral blood obtained before and after IMRT, and the data was expressed the ratio of after treatment to before treatment.Results: The total number of radiation courses and total exposure doses of radiation were similar between the control and H2 groups. IMRT with HCC therapy significantly reduced white blood cells (WBC) and platelets (PLT), but not red blood cells (RBC), hemoglobin (HGB) and hematocrit (HT). In contrast, H2 gas treatment significantly alleviates reducing effects of WBC and PLT (p=0.0011 and p=0.0275, respectively). Bone marrow damage index calculated by total exposure dose and WBC ratio also demonstrated the radioprotective effects of H2 gas (p=0.0231). Tumor responses to IMRT were similar between the two groups, and 4 of 7 (57%) patients in the control group and 7 of 16 (44%) patients in the H2 group achieved a complete response (CR) or partial response (PR). In addition, 3 of 16 (19%) patients in the H2 group achieved stable disease (SD).Conclusion: The results obtained demonstrated that H2 gas inhalation therapy alleviated IMRT-induced bone marrow damage without compromising the anti-tumor effects of IMRT. The present study suggests that this novel approach of H2 gas inhalation therapy may be applicable to IMRT-induced bone marrow damage in cancer patients.
Methods of Radiology Reports Generation and Patterns of Associated Musculoskeletal Symptoms: A Cross-Sectional Multi-Center Study
Authors: Dandan Omran Al , Hassan Ali , Alsaif Hind , Shammari Malak Al , Gari Danya , Krupinski Elizabeth
Published: March 2020

Objectives This study aimed to investigate the association between the methods used to generate radiology reports and the prevalence of musculoskeletal symptoms among radiologists. Additionally, we investigated the factors associated with the use of such methods. Methods An online survey was sent to radiologists practicing in hospitals in the Eastern Province of Saudi Arabia. The survey addressed demographic characteristics, work environment, and methods used to generate radiology reports and included an evaluation of musculoskeletal symptoms using the Nordic Musculoskeletal Questionnaire. Results were analyzed descriptively using the Chi-square test and logistic regression analysis. Results The survey was completed by 198 radiologists (111 men and 87 women), including residents (40.9%), specialists (27.3%), and consultants (31.8%). Most participants (71.2%) were aged below 40 years. In total, 140 (70.7%) participants had experienced musculoskeletal symptoms in the week preceding the survey. The most popular method used to generate radiology reports was typing using a keyboard, with 64.1% of participants using this method, followed by a handheld dictation device (49.5%). Radiologists with limited years of practice more frequently generated reports via keyboard typing, particularly those with <1-year of experience (35.8%). A multivariable logistic regression analysis revealed that radiologists who used handheld dictation devices were at higher risk of developing musculoskeletal symptoms. Conclusions Musculoskeletal symptoms are common among radiologists. Radiologists who use handheld dictation devices were particularly likely to report experiencing musculoskeletal symptoms. These devices were less commonly used among radiologists with fewer years of experience.
The miniaturization ligament performance probe (MLPP) system for analysis of the ligament strain pattern in three-dimensional motion of the ankle
Authors: Takeuchi Yoshitaka , Inokuchi Ryota , Takao Masato , Glazebrook Mark , Oliva Xavier , Yamazaki Takayuki , Kubo Maya , Lowe Danielle , Matsui Kentaro , Katakura Mai , Ozeki Satoru
Published: March 2020

Background Measuring strain patterns of joint ligaments in various positions helps in understanding their function. There are few studies on biomechanical properties of ankle ligaments, and the tensile properties of each ligament during motion have not been described because the existing biomechanical sensors are too big to insert into the ankle. Hypothesis/Purpose: This study aimed to verify the validity of a novel miniaturized ligament performance probe (MLPP) system in measuring the strain pattern of the anterior talofibular ligament (ATFL) during ankle motion. Methods The system is composed of a strain gauge (force probe), an amplifier unit, a display unit, and a logger, which are widely used industrially. Ten fresh-frozen through-the-knee lower extremity cadaveric specimens were used. The MLPP was sutured into the midsubstance of ATFL fibers. To measure the tensile force, a round metal disk (clock) (diameter 150 mm), with a 6-mm-diameter hole every 30°, was fixed on the plantar aspect of the foot. With a 1.2-Nm load applied to the ankle and subtalar joint complex, the ankle was manually moved from 15° dorsiflexion to 30° plantar flexion. The clock was rotated every 30° to measure the strain of the ATFL at each end point detected by the miniature force probe. During the motion from 15° dorsiflexion to 30° plantar flexion, the ATFL tensed near 20° plantar flexion, and the strain increased as the plantar flexion angle increased. The ATFL was maximally tensioned at 3 and 4 o’clock in the inversion position. Results In the elastic range where the ATFL can return to its original shape and length, tensile force was proportional to strain in all cases. All specimens showed almost similar strain patterns, with coefficients of variation ranging from zero to one in axial motion and from 0.05 to one in clock motion. Conclusion The MLPP system was effective in establishing the relationship between the limb position and ligament strain pattern of the small ligaments of the ankle.
Live Poultry Market Closure and Avian Influenza A (H7N9) infection in cities of China, 2013-2017: An ecological study
Authors: Chen Ying , Cheng Jian , Xu Zhiwei , Hu Wenbiao , Lu Jiahai
Published: March 2020

Background Previous studies have proven that the closure of live poultry markets (LPMs) was an effective intervention to reduce human risk of avian influenza A (H7N9) infection, but evidence is limited on the impact of scale and duration of LPMs closure on the transmission of H7N9. Method Five cities (i.e., Shanghai, Suzhou, Shenzhen, Guangzhou and Hangzhou) with the largest number of H7N9 cases in mainland China from 2013-2017 were selected in this study. Data on laboratory-confirmed H7N9 human cases in those five cities were obtained from the Chinese National Influenza Centre. The detailed information of LPMs closure (i.e., area and duration) was obtained from the Ministry of Agriculture. We used a generalized linear model with a Poisson link to estimate the effect of LPMs closure, reported as relative risk reduction (RRR). We used classification and regression trees (CARTs) to select and quantify the dominant factor of H7N9 infection. Results All five cities implemented the LPMs closure, and the risk of H7N9 infection decreased significantly after LPMs closure with RRR ranging from 0.80-0.93. Respectively, a long-term LPMs closure for 10-13 weeks elicited a sustained and highly significant risk reduction of H7N9 infection (RRR = 0.98). Short-time LPMs closure with 2 weeks in every epidemic did not reduce the risk of H7N9 infection (p>0.05). Partially closed LPMs in some suburbs contributed only 35% for reduction rate (RRR=0.35). Shenzhen implemented partial closure for first 3 epidemics (p>0.05) and all closure in the latest 2 epidemic waves (RRR=0.64). Conclusion Our findings suggest that LPMs all closure in whole city can be a highly effective measure comparing with partial closure (i.e. only urban closure, suburb and country remain open). Extend the duration of closure and consider permanently closing the LPMs will help improve the control effect. The effect of LPMs closure is greater than that of meteorology on H7N9 transmission.
“Side effects-part of the package” – A mixed methods approach to adverse events among patients with DR-TB under RNTCP Gujarat, India
Authors: Jakasania Arjunkumar Hardas , shringarpure Kalpita , Kapadia Dixit , Sharma Radhika , Mehta Kedar , Prajapati Arpit , Soundappan Kathirvel
Published: March 2020

Background Unsuccessful treatment outcomes among patients with drug resistant tuberculosis (DR-TB) are associated with poor treatment adherence due to high rates of adverse events (AEs). However, AEs are not systematically collected and managed under programmatic setting. The present study aims to assess the a) incidence and pattern of adverse events in first three months of DR-TB treatment; b) treatment seeking behaviour for AE management; and c) explore the challenges perceived by patients and healthcare providers in seeking treatment/reporting AEs. Methods This mixed methods study included all patients diagnosed and initiated on treatment during July-September 2018 at Ahmedabad DR-TB centre under RNTCP. The patients were interviewed telephonically and assessed for all AEs experienced by them during first three months of DR-TB treatment initiation. In-depth interviews and key-informant interviews were conducted among patients and DOTS supervisors, and programme staff (treatment supervisors, Medical Officer and District Program Manager). Results Total 207 AEs were reported by the 74 DR-TB patients who were interviewed. All patients experienced at least one AE during treatment. Incidence rate of AEs (experienced) was 3.11 (1 st month-4.6, 2 nd month-2.7, 3 rd month-2.02) per 100 person days of the 207 AEs, gastro-intestinal (59, 28.3%), ophthalmic (32, 15.4%) and otolaryngology (25, 11.9%) system related AEs were commonly experienced. Treatment was not sought in two-fifths of the AEs. Themes and subthemes related to challenges in treatment seeking or reporting of AEs were 1) Patient related-Misconceptions, accessibility and affordability of management, lack of counselling support, stigma and discrimination, past treatment experience; 2) Health system related- lack of guideline and training for AE management, poor coordination between hospital and tuberculosis centre.Conclusion The incidence of AEs was high among patients with DR-TB in the first three months of treatment and treatment seeking/reporting was low. Adequate health education and counselling of the patient should be done at initiation of treatment. An efficient real-time reporting and management of AE can be developed and tested for effective DR-TB control.
Haplotyping by linked-read sequencing (HLRS) of the genetic disease carriers for preimplantation genetic testing without a proband or relatives
Authors: Li Qing , Mao Yan , Li Shaoying , Du Hongzi , He Wenzhi , He Jianchun , Kong Lingyin , Zhang Jun , Liang Bo , Liu Jianqiao
Published: March 2020

Background: In order to mitigate the risk of allele dropout (ADO) and ensure the accuracy of preimplantation genetic testing for monogenic disease (PGT-M), it is necessary to construct parental haplotypes.. Typically, haplotype resolution is obtained by genotyping multiple polymorphic markers in both parents and a proband or a relative. Sometimes, single sperm typing, or tests on the polar bodies may also be useful. Nevertheless, this process is time-consuming. At present, there was no simple linkage analysis strategy for patients without affected relatives.Method: To solve this problem, we established a haplotyping by linked-read sequencing (HLRS) method without the requirement for additional relatives. First, the haplotype of the genetic disease carriers in the family was constructed by linked-read sequencing, and then the informative single nucleotide polymorphisms (SNPs) in upstream and downstream mutation region were selected to construct the embryo haplotype and to determine whether the embryo was carrying the mutation. Two families were selected to validate this method; one with alpha thalassemia and the other with NDP gene disorder.Results: The haplotyping by linked-read sequencing (HLRS) method was successfully applied to construct parental haplotypes without recruiting additional family members; the method was also validated for PGT-M. The mutation carriers in these families were sequenced by linked-read sequencing, and their haplotypes were successfully phased. Adjacent SNPs of the mutation gene were identified. The informative SNPs were chosen for linkage analyses to identify the carrier embryos. For the alpha thalassemia family, a normal blastocyst was transferred to the uterus and the accuracy of PGT-M was confirmed by amniocentesis at 16 weeks of gestation. Conclusions: Our results suggest that HLRS can be applied for PGT-M of monogenic disorders or de novo mutations where the mutations haplotype cannot be determined due to absence of affected relatives. Keywords: Preimplantation Genetic Testing for monogenic disease, Linked-read sequencing, Linkage analyses, Haplotype
Validation of the French version of the McLean Screening Instrument for Borderline Personality Disorder (MSI-BPD)
Authors: Pham-Scottez Alexandra , Mirkovic Bojan , Cailhol Lionel , Guelfi Julien-Daniel , Perez-Diaz Fernando , Speranza Mario , Corcos Maurice , Zanarini Mary
Published: March 2020

Background The study examines the psychometric properties of the French version of the McLean Screening Instrument for Borderline Personality Disorder (MSI-BPD) created by M. Zanarini to screen borderline personality disorder in clinical and non-clinical populations. Method In this multicentric longitudinal study from the European Network on Borderline Personality Disorder, a sample of 85 adolescent patients from five psychiatric centres and 85 matched controls without psychiatric comorbidity completed the MSI-BPD, French version, and were interviewed with the Structured Interview for DSM-IV Personality (SIDP-IV), in order to assess the presence or absence of borderline personality disorder. Results The MSI-BPD showed excellent internal consistency (α = 0.87 [0.84;0.90]). Compared to the semi-structured reference interview (SIDP-IV), the MSI-BPD showed substantial congruent validity (AUC = 0.93, CI 95%: 0.90 - 0.97). The optimal cut-off point in the present study was 5 or more as it had relatively high sensitivity (0.87) and specificity (0.85). In our sample, the cut-off point (7 or more) proposed by the original developers of the MSI-BPD showed high specificity (0.95) but low sensitivity (0.63). Conclusions The French version of the MSI-BPD is now available, and its psychometric properties are satisfactory. The French version of the MSI-PBD can be used as a screening tool for borderline personality disorder, for clinical purposes or in research studies.
High level of contamination of antimicrobial drug resistant non-typhoidal Salmonella enterica in commercial poultry and its surroundings in Chitwan, Nepal
Authors: Nelson Amy , Manandhar Sulochana , Ruzante Juliana , Gyawali Arrogya , Dhakal Bimala , Dulal Santosh , Chaulagai Rupendra , Dixit Sameer M
Published: March 2020

BackgroundAntimicrobial resistance (AMR) among bacterial pathogens is a fast-growing public health concern. AMR in non-typhoidal Salmonella species among food animals is of special concern as this may transmit resistant pathogens to humans during handling or consumption of animal products. In Nepal, the possibility of AMR Salmonella species among food animals is an important area of research, particularly in light of the rapidly growing poultry industry, lack of surveillance, and paucity of studies that have been conducted. MethodsTaking one health approach, a cross-sectional study was carried out in Chitwan district of Nepal between May and October 2017. Various environmental samples viz. farm litter, feed, water, poultry feces, vehicle swabs, farm swabs from 12 broiler poultry farms and various sections of poultry carcasses from 21 slaughter houses were aseptically collected. These were microbiologically assessed for the presence of non-typhoidal Salmonella and their phenotypic and genotypic indicators of antimicrobial resistance. ResultsOverall, Of 62 environmental samples collected, 31(50%) tested positive for Salmonella enterica serovars with environmental swabs (70%, 8/12) being the most culture positive sample types. Similarly, of 159 tissue samples collected from 24 carcasses, 79% (126/159) were culture positive for Salmonella enterica. Nearly 97% (153/157), 11% (17/157), 5% (8/157) and none of isolates showed resistance to tetracycline, ciprofloxacin, azithromycin and meropenem respectively. Some 83% (131/157), 40% (64/157), 8% (13/157) and 0.6% (1/157) of isolates tested positive for tetA, QrnS, mefA and VIM-1 AMR genes corresponding to the above antimicrobials respectively.ConclusionsThis study revealed gross contamination of farms and subsequent poultry meat samples with Salmonella enterica serovars that were resistant to several clinically applicable antimicrobials. This reinforces an urgent need to implement proper biosecurity approaches from farms to slaughter houses and strengthen policies to cease the rampant use of clinically important antimicrobials in poultry.
Transport of Goods Powered by Cereal Straw: Temporal and Spatial Biomass Availability for Future Bio-LNG Production in Germany
Authors: Brosowski André , Bill Ralf , Thrän Daniela
Published: March 2020

Background: Half of the UN climate target for 2030 has been achieved and further progress requires swiftly implementable solutions. In this context, the fermentation of cereal straw is a promising option. Returning the digestate to the farmland can close agricultural cycles while simultaneously producing biomethane for the transport sector. The world's first large-scale, mono-digestion plant for straw is operational since 2014. The temporal and spatial biomass availability is a key issue when replicating this concept. No detailed calculations on this subject are available, and the strategic relevance of biomethane from straw in the transport sector cannot be sufficiently evaluated.Methods: To assess the balance of straw supply and use, a total of 30 data sets are combined, taking into account the cultivation of the five most important cereal types and the straw required for ten animal species, two special crops and twelve industrial uses. The data are managed at district level and presented for the years 2010 to 2018. In combination with high-resolution geodata, the results are linked to actual arable fields, and the availability of straw throughout the country is evaluated using a GIS.Results: During the analysis period, the mobilisable potential for future biomethane production is between 13.9–21.5 Tg fm a-1; this is up to 62 % higher than the previously known level. The annual potential fluctuates considerably due to weather anomalies. The all-time maximum in 2014 and the minimum for the last 26 years in 2018 are separated by just four years and a difference of 7.6 Tg fm. However, large parts of the potential are concentrated only in a few regions and liquefied biomethane could fully cover the fuel required for vessels, and up to a quarter of that for heavy goods vehicles. Up to 11.3 Tg CO2-eq. could be saved, reducing the difference to achieve the UN climate target by 3.7 %.Conclusion: Despite the strong fluctuations, the potential is sufficient to supply numerous plants and to produce relevant quantities of liquefied biomethane even in weak years. To unlock the potential, the outcomes should be discussed further with stakeholders in the identified priority regions.
Nomograms Predict Overall Survival And Cancer-Speciﬁc Survival In Patients With Fibrosarcoma: A SEER-Based Study
Authors: Xiang Guangheng , Zhu Juan-Juan , Ke Chen-Rong , Weng Yi-Min , Fang Ming-Qiao , Zhu Si-Pin , Li Yu-An , Xiao Jian , Xu Lei
Published: March 2020

Background Due to the rarity, it is difficult to predict the survival of patients with fibrosarcoma. This study aimed to apply a nomogram to predict survival outcomes in patients with fibrosarcoma. Methods A total of 2235 patients with diagnoses of fibrosarcoma were registered in the Surveillance, Epidemiology, and End Results database, of whom 663 patients were eventually enrolled. Univariate and multivariate Cox analyses were used to identify independent prognostic factors. Nomograms were constructed to predict 3‐ and 5‐year overall survival and cancer‐specific survival of patients with fibrosarcoma. Results In univariate and multivariate analyses of OS, age, sex, race, tumor stage, pathologic grade, use of surgery and tumor size identiﬁed as independent prognostic factors. Age, sex, tumor stage, pathologic grade, use of surgery and tumor size were significantly associated with CSS. These characteristics were further included to establish the nomogram for predicting 3- and 5- year OS and CSS. For the internal validation of the nomogram predictions of OS and CSS, the C-indices were 0.784 and 0.801. Conclusion We developed and validated the nomograms that estimated 3- and 5-year OS and CSS. These nomograms not only have good discrimination performance and calibration, but also provide patients with better clinical benefits.
Primary tumor location affects recurrence-free survival for patients with colorectal liver metastases after hepatectomy: A propensity score matched analysis
Authors: Zhang Yuanping , Wang Yongjin , Yuan Yichuan , Qiu Jiliang , Qiu Yuxiong , He Wei , Zheng Yun , Wang Zhiqiang , Gu Yangkui , Lu Zhenhai , Chen Gong , Ding Peirong , Wu Xiaojun , Pan Zhizhong , Wan Desen , Li Yuhong , Xu Ruihua , Yuan Yunfei , Li Binkui
Published: March 2020

Background Whether primary tumor location of colorectal cancer (CRC) affects survival of patients after resection of liver metastases remains controversial. This study was conducted to investigate the differences in clinicopathological characteristics and prognosis between right-sided CRC and left-sided CRC patients with liver metastases after hepatectomy. Methods From 2002 to 2018, 611 patients with colorectal liver metastases (CRLM) who underwent hepatectomy at our center were reviewed. Primary tumors located from cecum to transverse colon were defined as right-sided group (n = 141); tumors located from splenic flexure to rectum were defined as left-sided group (n = 470). Patients were compared between two groups before and after a 1:1 propensity score analysis (PSM). Results Before PSM, median survival time and 5-year overall survival (OS) rate in right-sided group were 77 months and 56.3%, and those in left-sided group were 64 months and 51.1%, respectively. After PSM, median survival time and 5-year OS rate in right-sided group were 77 months and 55.9%, and those in left-sided group were 58.8 months and 47.3%, respectively. The OS rates did not differ between two groups before and after PSM ( P = 0.575; P = 0.453). However, significant different recurrence free survival (RFS) rate was found before and after PSM between right-sided and left-sided group ( P = 0.028, P = 0.003). Conclusions Primary tumor location of CRC impacts RFS for patients with liver metastases after resection. A more frequent follow up to detect early recurrence might be justified for CRLM patients with a right-sided CRC.
Systemic treatment with human amnion epithelial cells after experimental traumatic brain injury
Authors: Kim Hyun Ah , Semple Bridgette D , Dill Larissa K , Pham Louise , Dworkin Sebastian , Zhang Shenpeng R , Lim Rebecca , Sobey Christopher G , McDonald Stuart J
Published: March 2020

Background: Systemic administration of human amnion epithelial cells (hAECs) was recently shown to reduce neuropathology and improve functional recovery following ischemic stroke in both mice and marmosets. Given the significant neuropathological overlap between ischemic stroke and traumatic brain injury (TBI), we hypothesized that a similar hAEC treatment regime would also improve TBI outcomes. Methods: Male mice (12 weeks old, n=40) were given a sham injury or moderate severity TBI by controlled cortical impact. At 60 minutes post-injury, mice were given a single tail vein injection of either saline (vehicle) or 1x10 6 hAECs suspended in saline. At 24 h post-injury, mice were assessed for locomotion and anxiety using an open field, and sensorimotor ability using a rotarod. At 48 h post-injury, brains were collected for analysis of immune cells via flow cytometry, or histological evaluation of lesion volume and hAEC penetration. To assess the impact of TBI and hAECs on lymphoid organs, spleen and thymus weights were determined. Results: Treatment with hAECs did not prevent TBI-induced sensorimotor deficits at 24 h post-injury. hAECs were detected in the injured brain parenchyma; however, lesion volume was not altered by hAEC treatment. Robust increases in several leukocyte populations in the ipsilateral hemisphere of TBI mice were found when compared to sham mice at 48 h post-injury; however, hAEC treatment did not alter brain immune cell numbers. Both TBI and hAEC treatment were found to increase spleen weight. Conclusions: Taken together, these findings indicate that — unlike in ischemic stroke — treatment with hAEC was unable to prevent immune cell infiltration and sensorimotor deficits in the acute stages following controlled cortical impact in mice. Although further investigations are required, our data suggests that the lack of hAEC-induced neuroprotection in the current study may be explained by the differential splenic contributions to neuropathology between these brain injury models.
MAGE-A4, NY-ESO-1 and SAGE expression rates and co-expression relationships in solid tumours
Authors: Ishihara Mikiya , Kageyama Shinichi , Miyahara Yoshihiro , Ishikawa Takeshi , Ueda Shugo , Soga Norihito , Naota Hiroaki , Mukai Katsumi , Harada Naozumi , Ikeda Hiroaki , Shiku Hiroshi
Published: March 2020

Background

Cancer testis (CT) antigens are promising targets for cancer immunotherapies such as cancer vaccines and genetically modified adoptive T cell therapy. In this study, we evaluated the expression of three CT antigens, melanoma-associated antigen A4 (MAGE-A4), New York oesophageal squamous cell carcinoma 1 (NY-ESO-1) and sarcoma antigen gene (SAGE).

Methods

MAGE-A4, NY-ESO-1 and/or SAGE antigen expression in tumour samples was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Informed consent was obtained from individuals prior to study enrolment.

Results

In total, 585 samples in 21 tumour types were evaluated between June 2009 and March 2018. The positive expression rates of these CT antigens were as follows: MAGE-A4, 34.6% (range, 30.7-38.7); NY-ESO-1, 21.0% (range, 17.2-25.1); and SAGE, 21.8% (range, 18.5-25.4). The MAGE-A4 antigen was expressed in 54.9% of oesophageal cancers, 37.5% of head and neck cancers, 35.0% of gastric cancers and 34.2% of ovarian cancers; the NY-ESO-1 antigen was expressed in 28.6% of lung cancers, 25.3% of oesophageal cancers and 22.6% of ovarian cancers; and the SAGE antigen was expressed in 35.3% of prostate cancers, 32.9% of oesophageal cancers and 26.3% of ovarian cancers. The most common tumour type was oesophageal cancer in this study. MAGE-A4, NY-ESO-1 and SAGE antigen expression were assessed in 214 oesophageal cancer samples, among which 24 (11.2%) were triple-positive, 58 (27.1%) were positive for any two, 59 (27.6%) were positive for any one, and 73 (34.1%) were triple-negative.

Conclusions

Oesophageal cancer exhibited a relatively high rate of CT antigen positivity. Our data indicate that oesophageal cancer is a promising candidate tumour for CT antigen-targeting immunotherapy and immune monitoring.
Identification of a novel methylation gene sites signature to predict prognosis in kidney renal papillary cell carcinoma by comprehensive profiling
Authors: wang yinglei , Zhang YingYing
Published: March 2020

Most kidney renal papillary cell carcinoma (KIRP) patients are diagnosed at the advanced stage and have a poor prognosis. DNA methylation plays an important role in prognosis prediction of cancers. Identification of novel DNA methylation site biomarkers could be beneficial for the prognosis of KIRP patients. In this study, we aimed to find an efficient methylated sites model for predicting survival in KIRP.
The safety and effectiveness of MIDLF during one surgeon’s initial phase: accuracy, clinical outcome, complications and learning curve
Authors: Ding Hongtao , Hai Yong , Liu Yuzeng , Guan Li , Pan Aixing , Han Bo
Published: March 2020

ObjectiveTo characterize the safety and effectiveness during one surgeon’s initial phases of performing a Midline lumbar fusion by freehand technique based on perioperative parameters.MethodsThe first 90 consecutive patients underwent single level MIDLF, utilized a consistent technique, performed by one surgeon were retrospectively studied. A piecewise regression analysis was performed to evaluate the operation duration based on case number, and to estimate the breakpoint. The early learning period and the late steady phase was divided according to the case number of the breakpoint. Perioperative parameters and clinical data of 2 groups were compared then to elucidate if improvement in operative duration was accompanied by better patient prognosis.ResultsThe inflectional asymptote of the surgeon’s learning curve for MIDLF was achieved at the 40th case. The demographics parameters were similar between the early group of 40 patients and the latter 50. Average surgical time, intraoperative blood loss, length of incision, duration of drainage, postoperative drainage and post-operative hospitalization were significantly less in the late cohort (p<.05). Medial cortical breach was observed in 6/360 cases (4 vs. 2, P>.05), and lateral cortical breach was observed in 35/360 cases (24 vs.11, P<.05). One case of cerebrospinal fluid (CSF) leak, two cases of durotomy and one case of nerve root injury related to the technique was found in the early group, and only one case of durotomy was found in the late group. Post-surgical wound infection was seen in 2.22% (N = 2/90) of patients (2 vs. 0, P>.05). The overall complication incidence decreased alone with the learning curve, and statistical difference was achieved. Furthermore, there was no difference in clinical improvement between the two groups.ConclusionsOur result indicated that technical proficiency in MIDLF was achieved after 40 surgeries. And it is effective to significantly improve the patients’ functional status and relief the pain during one surgeon’s learning process of MIDLF in the treatment of lumbar degenerative disease, even in the initial phase. As the surgeon gains surgical proficiency and familiarity with MIDLF procedure, potential complications could be minimized. Furthermore, our results provided evidence to the excellent performance as an alternative option to traditional pedicle screw fixation.