bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of efficiency of in vitro cloned sheep embryo production by conventional somatic cell nuclear transfer and handmade cloning technique
Authors: Khan S , Tali M , Khan A , Bhat S , Ashraf A , Bhat MH , Khan F , Shah RA
Published: January 2018

Contents

Conventional somatic cell nuclear transfer (SCNT) technique of in vitro production of cloned embryos involves use of costly and complicated micromanipulators. Handmade cloning (HMC) technique has been applied as efficient and cost‐effective alternative in many livestock species. The aim of the present study was to compare the efficiency of in vitro production and in vitro development of cloned sheep embryos by the two techniques. Cloned embryos were produced by conventional SCNT using micromanipulator apparatus and by HMC technique. Enucleation efficiency and efficiency of fusion with somatic cell (nucleus donor) were compared. Cleavage percentage was observed on day 2 of in vitro culture (IVC), and morula and blastocyst percentages were calculated on day 7 of IVC. Higher enucleation efficiency (96.98 ± 1.01 vs. 93.62 ± 1.03; p > .05) as well as fusion efficiency was obtained with HMC technique than with conventional SCNT (96.26 ± 1.34 vs. 92.63 ± 0.70, p < .05); 181 cloned sheep embryos were produced in vitro by conventional SCNT and 92 by HMC. Cleavage percentage observed on day 2 of in vitro culture was higher in HMC than SCNT (66.92 ± 3.72 vs. 55.97 ± 2.5, respectively, p < .05). Morula percentage obtained was higher in SCNT than HMC (44.12 ± 2.93 vs. 30.43 ± 6.79, respectively, p < .05), whereas blastocyst percentage obtained by HMC was higher (12.46 ± 4.96) than SCNT (5.31 ± 2.25; p > .05). It was inferred that HMC technique provides a cost‐effective and efficient method of in vitro production of cloned sheep embryos with a comparatively simpler technique with a possibility of automation. Efficiency of cloned embryo production could be improved further by propagating and standardizing this technique.
Quantifying soil displacement and tillage erosion rate by different tillage systems in dryland northwestern Iran
Authors: Kouselou M. , Hashemi S. , Eskandari I. , McKenzie B. M. , Karimi E. , Rezaei A. , Rahmati M.
Published: January 2018

Redistribution of soil particles by the force applied by tillage is a major factor in soil degradation of agricultural land. Decreasing tillage intensity can reduce the amount of soil displaced and the distance moved and hence may reduce rates of erosion. To understand the relative importance of erosion, we tabulated machine, soil and landform properties likely to be involved. We compared soil displacement and tillage erosion rates under different systems, including mouldboard ploughing (conventional tillage), chisel ploughing (reduced tillage), stubble cultivator (minimum tillage) and no‐tillage under dryland agriculture in northwestern Iran. The area was undulating and so all tillage took place along contours. Metallic tracers were buried in the soil at known locations and depths and their recovery after tillage provided a measure of soil displacement and tillage erosion. Conventional tillage along a contour line caused significantly greater soil displacement (≃57 cm) in the direction of tillage than reduced and minimum tillage systems (~20 and ~15 cm, respectively). Conventional tillage also caused more lateral soil displacement (downwards in the main direction of slope, the tillage erosion rate) than reduced or minimum tillage systems (48 cm or 152 kg/m vs. 5 and 4 cm or 16 and 7 kg/m, respectively). Although a range of factors contribute to the tillage systems used by farmers, our results suggest that under dryland conditions, similar to those found in our study area, adoption of noninversion, reduced tillage along the contour, for example by chisel ploughing can substantially limit tillage erosion relative to conventional tillage.
The late Messinian event: A worldwide tectonic revolution
Authors: Leroux Estelle , Aslanian Daniel , Rabineau Marina , Pellen Romain , Moulin Maryline
Published: January 2018

A review of geological and geophysical observations points towards a worldwide kinematic change at around 6 Ma. The synchronicity of many manifestations (tectonics, magmatism, kinematics, ecological events, among others) at ~6 Ma, similar to those recognized from time to time on the geological time‐scale, argues for a global geodynamic event that has led to many regional consequences on Earth's surface. In particular, we propose that this global event was the main trigger for the three fold increase in sediment deposits in the world ocean over the last ~5 Ma, but also for the onset of the Messinian Salinity Crisis in the Mediterranean area, one of the most severe ecological crises in the Earth's history. We suggest this Messinian revolution to be the last occurrence of cyclic successions of global events.
Association between the time after transplantation and different immunosuppressive medications with dental and periodontal treatment need in patients after solid organ transplantation
Authors: Schmalz Gerhard , Wendorff Horst , Berisha Lisa , Meisel Anja , Widmer Florian , Marcinkowski Anna , Teschler Helmut , Sommerwerck Urte , Haak Rainer , Kollmar Otto , Ziebolz Dirk
Published: January 2018

Objective

The aim of this study was to investigate the association of time after transplantation and different immunosuppressive medications with dental and periodontal treatment needs in patients after solid organ transplantation (SOT).

Methods

After lung, liver, or kidney transplantation, patients were included and divided into subgroups based on the time after SOT (0‐1, 1‐3, 3‐6, 6‐10, and >10 years) and immunosuppression (tacrolimus, cyclosporine, mycophenolate, glucocorticoids, sirolimus, and monotherapy vs combination). Dental treatment need was determined by the presence of carious lesions, while periodontal treatment need was diagnosed based on a Periodontal Screening index score of 3‐4. The overall treatment need included both the dental and/or periodontal treatment needs. Statistical analysis was performed using the Kruskal‐Wallis test and chi‐squared test (P < .05).

Results

A total of 169 patients were included after SOT. A dental treatment need of 44%, a periodontal treatment need of 71%, and an overall treatment need of 84% were detected in the total cohort. Only patients with >10 years after SOT had a lower dental treatment need compared to the other groups (P = .02). All other comparisons of dental, periodontal, and overall treatment needs were comparable between subgroups depending on time since SOT. Furthermore, no statistically significant differences were found in terms of the dental, periodontal, or overall treatment needs following the administration of different immunosuppressive medications.

Conclusion

The high treatment need of patients after SOT, irrespective of the time since transplantation, suggests insufficient dental and periodontal treatment before and maintenance after organ transplantation. Furthermore, immunosuppressive medication was not associated with the treatment need.
Early infections in the intensive care unit after liver transplantation‐etiology and risk factors: A single‐center experience
Authors: Laici Cristiana , Gamberini Lorenzo , Bardi Tommaso , Siniscalchi Antonio , Reggiani Maria Letizia Bacchi , Faenza Stefano
Published: January 2018

Background

Infectious complications represent one of the main causes of perioperative morbidity and mortality of liver transplant recipients. The primary objective of this retrospective observational study was to evaluate incidence and etiology of early (within 1 month from surgery and occurring in the intensive care unit [ICU]) postoperative infections as well as donor‐ and recipient‐related risk factors.

Methods

The data of 280 patients undergoing 299 consecutive liver transplant procedures from January 2012 to December 2015 were extracted from the Italian ICU registry database and hospital registries. Perioperative risk factors, etiology of infections, and antibiotic susceptibility of isolated microorganisms were taken into consideration.

Results

Global incidence of postoperative infections was 21%. Pneumonia was the most frequent infection and, globally, gram‐negative bacteria were the most common agents. Septic shock was present in 22% of infection cases and hospital mortality was higher in patients with postoperative infection. Preoperative chronic obstructive pulmonary disease, malnutrition, preoperative ascites, encephalopathy, and early re‐transplantation were significantly associated to post orthotopic LT infections.

Conclusion

Infections represent a major cause of early postoperative morbidity and mortality. The impact of single risk factors and the results of their preoperative management should be further investigated in order to reduce the incidence and evolution of postoperative infections.
Carbapenem‐resistant Klebsiella pneumoniae vertebral osteomyelitis in a renal transplant recipient treated with ceftazidime‐avibactam
Authors: Cani Eris , Moussavi Farzad , Ocheretyaner Eric , Sharma Roopali , Brown Clinton , Eilertson Brandon
Published: January 2018

Ceftazidime‐avibactam (CAZ‐AVI) is a novel cephalosporin beta lactamase inhibitor combination that has shown activity against carbapenem‐resistant Enterobactericeae. Data are limited on its utilization in the treatment of carbapenem‐resistant Klebsiella pneumoniae osteomyelitis in solid organ transplant patients. We describe a case report on the use of CAZ‐AVI in the treatment of vertebral osteomyelitis in a renal transplant recipient.
A successful aggressive surgical and medical approach to pulmonary artery obstruction due to Mycobacterium abscessus infection post lung transplantation
Authors: Kotecha Sakhee , Smibert Olivia C. , Doi Atsuo , Gooi Julian , Morrissey Orla , Paraskeva Miranda , Snell Greg I. , McGiffin David
Published: January 2018

Mycobacterium abscessus infection following lung transplantation has historically been associated with poor outcomes. We report a case of bilateral lung retransplantation complicated by obstruction of the right pulmonary artery secondary to M. abscessus mycotic aneurysm. Aggressive surgical management, including reconstruction of the right pulmonary artery, was undertaken with prolonged antimicrobial therapy. Thirty‐six months later, antibiotics have been discontinued and the patient has stable soft tissue chest wall disease with good graft function. Mortality and morbidity associated with M. abscessus infection is considerable but this case illustrates that with aggressive early management, outcomes may be favorable.
Successful resolution of hyperammonemia following hematopoietic cell transplantation with directed treatment of Ureaplasma parvum infection
Authors: Graetz Riley , Meyer Robyn , Shehab Kareem , Katsanis Emmanuel
Published: January 2018

Hyperammonemia following hematopoietic cell transplantation (HCT) has been characterized as idiopathic and is associated with a very high mortality. A causal relationship between Ureaplasma infection and hyperammonemia in immunocompromised lung transplant recipients has recently been described. We document the first case of hyperammonemia following HCT associated with Ureaplasma parvum. The initiation of appropriate antibiotics resulted in rapid resolution of hyperammonemic encephalopathy and eradication of the implicating organism.
Long‐term follow‐up of post renal transplantation Epstein‐Barr virus‐associated smooth muscle tumors: Report of two cases and review of the literature
Authors: El Hennawy Hany M. , Habhab Wael , Almutawa Abdulmonem , Shinawi Sumaih , Al Ayad Abdulrahman , Fahmy Ahmed
Published: January 2018

Epstein‐Barr virus (EBV)‐associated smooth muscle tumors (SMTs) following solid organ transplantation are very rare slow growing neoplasms. Most tumors present with non‐specific symptoms mainly related to tumor location. Post‐transplant EBV‐associated small muscle tumors have been reported in various anatomical locations. The tumors have a predilection to unusual sites for SMTs and tend to be multifocal. The histologic appearance of these tumors generally does not predict their clinical behavior. Surgery and reduction in immunosuppression are the main stays of management. We herein report two cases of post renal transplant EBV‐associated SMTs with over 6 years of follow‐up. A 33‐year‐old male patient presented with hepatic lesions and a 49‐year‐old female patient presented with multiple mesenteric and gluteal lesions. The tumors were diagnosed 6 and 10 years after renal transplantation, respectively. Surgical resection and reduction/discontinuation of immunosuppression were successful in delaying progression of the disease; however, in both cases, the allografts failed during the course of management.
Hepatitis A, cardiomyopathy, aplastic anemia, and acute liver failure: A devastating scenario
Authors: Botero Verónica , García Víctor H. , Aristizabal Ana M. , Gomez Catalina , Perez Paola , Caicedo Luis A. , Echeverri Gabriel J.
Published: January 2018

Hepatitis A virus (HAV) causes an acute infection and is usually asymptomatic in children. When clinical manifestations appear, these include choluria, jaundice, and abdominal pain. Although infrequent, extra‐hepatic manifestations related to HAV have been described, affecting the heart, bone marrow, blood vessels, and other tissues.A 10‐year‐old boy from a rural area presented with a 15‐day history of malaise, fever, and jaundice; laboratory examinations were compatible with HAV infection. The patient turned encephalopathic and was remitted to our center, where laboratory examinations showed a medullary aplasia and fulminant hepatitis requiring a liver transplant that was performed 72 hours after admission. At 24 hours post transplant, the patient developed a cardiomyopathy secondary to HAV, and intravenous immunoglobulin was administered. The patient is still alive and attending his medical check‐ups.Although rare, extra‐hepatic manifestations of HAV infection have been described in 14% of cases. The groups of patients affected are usually aged and present with high bilirubin levels. Acquired aplastic anemia and myocarditis caused by HAV are uncommon, and its pathophysiology has not yet been elucidated.HAV infection is usually asymptomatic in children, although extra‐hepatic manifestations can appear requiring early detection and management.
Acanthamoeba endophthalmitis during treatment for cutaneous disease in a renal transplant patient
Authors: Kutner Allison , Aldrich Margaret , Patel Sarju , Kang Joann J. , Amin Bijal , Mann Ranon , Ali Ibne Karim M. , Martines Roosecelis Brasil , Cope Jennifer Rittenhouse , De Boccardo Graciela O. , Puius Yoram A.
Published: January 2018

Acanthamoeba infections are difficult to diagnose and treat. We present a renal transplant patient who developed Acanthamoeba endophthalmitis on therapy with posaconazole and miltefosine for cutaneous acanthamobiasis. The patient was maintained on intracameral voriconazole injections, and oral azithromycin, fluconazole, and flucytosine. This case highlights novel presentations and treatments for acanthamoebic infection.
Comparative effectiveness of aerosolized versus oral ribavirin for the treatment of respiratory syncytial virus infections: A single‐center retrospective cohort study and review of the literature
Authors: Trang Tracy P. , Whalen Meghan , Hilts‐Horeczko Alexandra , Doernberg Sarah B. , Liu Catherine
Published: January 2018

Background

Respiratory syncytial virus (RSV) is a leading cause of viral infections in immunocompromised hosts and is associated with significant morbidity and mortality. In January 2015, our institution switched from aerosolized to oral ribavirin (RBV) for primary treatment of RSV infection among high‐risk immunocompromised adult patients. The objective of the study was to evaluate the clinical and economic outcomes associated with this switch.

Methods

Retrospective cohort analysis of adult patients diagnosed with RSV infection and treated with RBV between January 1, 2013, and May 31, 2016.

Results

Of 46 patients, 20 (43%) patients received oral RBV and 26 (57%) received aerosolized RBV. Underlying conditions included lung transplant (n = 22), hematopoietic cell transplant (n = 16), hematological malignancy (n = 5), and structural lung diseases (n = 4). At the time of RSV diagnosis, 42 (91%) were hospitalized, 36 (78%) had clinical and radiographic evidence of lower respiratory tract infection (RTI), 9 (20%) were admitted to the ICU, and 4 (9%) required intubation. There were no differences in clinical outcomes between the 2 groups with regard to adverse events, progression from upper to lower RTI, escalation of care, or 30‐day mortality. Three (15%) in the oral group and 1 (4%) in the aerosolized group died within 30 days (P = .33). The cost avoidance attributable to the switch in therapy over 1 year is $1.2 million.

Conclusion

Oral RBV appears to be a safe and cost‐effective alternative to aerosolized RBV for the management of RSV infection in immunocompromised patients. Larger studies are needed to validate the safety and efficacy of this approach.
Tuberculosis after liver transplantation in a large center in New York City: QuantiFERON®‐TB Gold‐based pre‐transplant screening performance and active tuberculosis post‐transplant
Authors: Hand Jonathan , Sigel Keith , Huprikar Shirish , Hamula Camille , Rana Meena
Published: January 2018

Introduction

Pre‐transplant screening for latent tuberculosis infection (LTBI) is a complex consideration that varies by institution. Inconsistent performance of interferon‐gamma release assay (IGRA) further complicates screening. Data regarding LTBI screening outcomes and test characteristics in a large, foreign‐born pre‐transplant population within the United States are limited.

Methods

In this retrospective study, patients who received QuantiFERON®‐TB Gold (QFT) prior to liver transplantation (LT) were included. Characteristics of patients were compared by QFT result, and predictors of indeterminate results were evaluated. Similar comparisons were performed between patients who developed active TB and those who did not.

Results

Of 148 patients screened, the rate of positive, indeterminate, and negative testing was 13.5% (20/148), 27% (40/148), and 59% (88/148), respectively. An indeterminate QFT result was more than 16 times more likely in patients with a Model for End‐stage Liver Disease score >25 (odds ratio [OR] 16.7; 95% confidence interval [CI], 2.1‐132.0; P = .008) and more than 4 times when performed in our institution's lab compared with commercial lab (OR 4.1; 95% CI, 1.34‐12.44; P = .013). The overall TB incidence was 1102/100 000 transplant cases. No patient who developed active TB had a positive QFT. All were born outside of the United States (P = .06) and had pre‐transplantation chest imaging demonstrating granulomatous disease (P = .006).

Conclusion

Our experience further highlights the challenges of LTBI screening prior to LT and suggests that QFT may be a poor predictor of active TB in higher risk pre‐transplant populations. Candidates should be screened as early as possible to optimize QFT performance, and local epidemiological data should be used to create institution‐specific screening protocols in areas with large populations from TB‐endemic regions. Management should consider TB risk factors, QFT, and imaging instead of reliance on QFT testing alone.
Progressive multifocal leukoencephalopathy after T‐cell replete HLA‐haploidentical transplantation with post‐transplantation cyclophosphamide graft‐versus‐host disease prophylaxis
Authors: Ikegawa Shuntaro , Fujii Nobuharu , Tadokoro Koh , Sato Kota , Iwamoto Miki , Matsuda Masayuki , Inomata Tomoko , Sugiura Hiroyuki , Asano Takeru , Yoshida Shohei , Nishimori Hisakazu , Matsuoka Ken‐ichi , Maeda Yoshinobu
Published: January 2018

A 52‐year‐old man suffered from progressive multifocal leukoencephalopathy (PML) after human leukocyte antigen (HLA)‐haploidentical transplantation with post‐transplantation cyclophosphamide (PTCY). Mirtazapine, mefloquine, and cytarabine failed to improve his symptoms, and he finally died 4.5 months after PML onset. This is the first case report of a patient with PML after HLA‐haploidentical transplantation with PTCY. Although T‐cell replete HLA‐haploidentical transplantation with PTCY has enabled early immune reconstitution, PML should be considered if a patient's mental condition deteriorates.
Discrimination between the endoplasmic reticulum and mitochondria by spontaneously inserting tail‐anchored proteins
Authors: Figueiredo Costa Bruna , Cassella Patrizia , Colombo Sara F. , Borgese Nica
Published: January 2018

Tail‐anchored (TA) proteins insert into their target organelles by incompletely elucidated posttranslational pathways. Some TA proteins spontaneously insert into protein‐free liposomes, yet target a specific organelle in vivo. Two spontaneously inserting cytochrome b5 forms, b5‐ER and b5‐RR, which differ only in the charge of the C‐terminal region, target the endoplasmic reticulum (ER) or the mitochondrial outer membrane (MOM), respectively. To bridge the gap between the cell‐free and in cellula results, we analyzed targeting in digitonin‐permeabilized adherent HeLa cells. In the absence of cytosol, the MOM was the destination of both b5 forms, whereas in cytosol the C‐terminal negative charge of b5‐ER determined targeting to the ER. Inhibition of the transmembrane recognition complex (TRC) pathway only partially reduced b5 targeting, while strongly affecting the classical TRC substrate synaptobrevin 2 (Syb2). To identify additional pathways, we tested a number of small inhibitors, and found that Eeyarestatin I (ESI) reduced insertion of b5‐ER and of another spontaneously inserting TA protein, while not affecting Syb2. The effect was independent from the known targets of ESI, Sec61 and p97/VCP. Our results demonstrate that the MOM is the preferred destination of spontaneously inserting TA proteins, regardless of their C‐terminal charge, and reveal a novel, substrate‐specific ER‐targeting pathway.
Macroeconomic policy cooperation and the G20
Authors: Triggs Adam
Published: January 2018

Has the G20 achieved its goals in macroeconomic cooperation since 2008? The paper organises the G20's efforts under five themes: macroeconomic stimulus, fiscal consolidation, monetary policy, the global financial safety net and global imbalances. The G20 was initially successful in each of these areas, but this success was short‐lived. While the G20 met its goals on macroeconomic stimulus, it has been less successful in reducing deficits and debt. While it was successful in increasing its resources, the global financial safety net remains too small, too fragmented and institutional reform is incomplete. While the G20 succeeded in moving to more market‐determined exchange rates and avoiding competitive devaluations, it struggled to avoid negative spillovers. Despite years of effort, the G20 has made limited progress in reducing global imbalances. Current account imbalances are creeping back to pre‐crisis levels. Public debt remains high and most economies are moving in the wrong direction in correcting imbalances in household savings and debt. The paper concludes that the G20 has done better in some areas than others. But to suggest the G20 is a forum in decline ignores its shift from reactive crisis response to longer‐term structural challenges outside of the pressing need of an immediate crisis.
Capnocytophaga keratitis in dogs: clinical, histopathologic, and microbiologic features of seven cases
Authors: Ledbetter Eric C. , Franklin‐Guild Rebecca J. , Edelmann Michele L.
Published: January 2018

Objective

To describe the clinical, microbiologic, and histopathologic features of Capnocytophaga keratitis in dogs.

Animals studied

Seven dogs with naturally acquired Capnocytophaga keratitis.

Procedures

Medical records of dogs with a clinical diagnosis of keratitis and corneal cultures positive for Capnocytophaga spp. were reviewed. Dog signalment, medical history, clinical findings, and diagnostic assay results were recorded.

Results

Breeds included Boston terrier (n = 3 dogs), Rat terrier (n = 2), and single cases of mixed breed and Pug. All dogs examined had expansive corneal ulceration involving the majority of the corneal surface. Marked corneal infiltrates, keratomalacia, and hypopyon were present. Progression of corneal disease was rapid with extensive dissolution of the corneal stroma. Corneal lesions progressed to catastrophic perforations within 24 h of the initial examination in three dogs, requiring enucleation. One globe was enucleated after failure to resolve with long‐term medical therapy. Globes and vision were retained in three dogs following aggressive medical therapy (two dogs) or 360° conjunctival graft surgery (one dog). Capnocytophaga cynodegmi, Capnocytophaga canimorsus, or unspeciated Capnocytophaga spp. were cultured from corneal samples of all dogs. Long, thin, gram‐negative rods were present during cytological evaluation of the cornea in some dogs. Histopathologic evaluation of enucleated globes revealed severe and diffuse neutrophilic and collagenolytic keratitis.

Conclusions

Capnocytophaga keratitis is a severe, rapidly progressive corneal infection in dogs that is associated with diffuse corneal involvement, extensive keratomalacia, and a relatively poor prognosis. Clinical features of canine Capnocytophaga keratitis are similar to human cases of this infection.
P‐ and E‐selectin receptor antagonism prevents human leukocyte adhesion to activated porcine endothelial monolayers and attenuates porcine endothelial damage
Authors: Laird Christopher T. , Hassanein Wessam , O'Neill Natalie A. , French Beth M. , Cheng Xiangfei , Fogler William E. , Magnani John L. , Parsell Dawn , Cimeno Arielle , Phelps Carol J. , Ayares David , Burdorf Lars , Azimzadeh Agnes M. , Pierson Richard N.
Published: January 2018

Background

Alongside the need to develop more effective and less toxic immunosuppression, the shortage of human organs available for organ transplantation is one of the major hurdles facing the field. Research into xenotransplantation, as an alternative source of organs, has unveiled formidable challenges. Porcine lungs perfused with human blood rapidly sequester the majority of circulating neutrophils and platelets, which leads to inflammation and organ failure within hours, and is not significantly attenuated by genetic modifications to the pig targeted to diminish antibody binding and complement and coagulation cascade activation.

Methods

Here, we model the interaction of freshly isolated human leukocytes with xenotransplanted vasculature under physiologic flow conditions using microfluidic channels coated with porcine endothelial cells. Both isolated human neutrophils and whole human blood were perfused over transgenic pig aortic endothelial cells that had been activated with rhTNF‐α or rhIL‐4 using the BioFlux system. Novel compounds GMI‐1271 and rPSGL1.Fc were tested as E‐ and P‐ selectin antagonists, respectively. Cellular adhesion and rolling events were tracked using FIJI (imageJ).

Results

Porcine endothelium activated with either rhTNF‐α or rhIL‐4 expressed high amounts of selectins, to which isolated human neutrophils readily rolled and tethered. Both E‐and P‐selectin antagonism significantly reduced the number of neutrophils rolling and rolling distance in a dose‐dependent manner, with near total inhibition at higher doses (P < .001). Similarly, with whole human blood, selectin blocking compounds exhibited dose‐dependent inhibition of prevalent leukocyte adhesion and severe endothelial injury (Untreated: 394 ± 97 PMNs/hpf, 57 ± 6% loss EC; GMI1271+rPSGL1.Fc: 23 ± 9 PMNs/hpf, 8 ± 6% loss EC P < .01).

Conclusions

Selectin blockade may be useful as part of an integrated strategy to prevent neutrophil‐mediated organ xenograft injury, especially during the early time points following reperfusion.
Strong xenoprotective function by single‐copy transgenes placed sequentially at a permissive locus
Authors: Rieblinger Beate , Fischer Konrad , Kind Alexander , Saller Benedikt S. , Baars Wiebke , Schuster Marion , Wolf‐van Buerck Lelia , Schäffler Andrea , Flisikowska Tatiana , Kurome Mayuko , Zakhartchenko Valeri , Kessler Barbara , Flisikowski Krzysztof , Wolf Eckhard , Seissler Jochen , Schwinzer Reinhard , Schnieke Angelika
Published: January 2018

Background

Multiple xenoprotective transgenes are best grouped at a single locus to avoid segregation during breeding and simplify production of donor animals.

Methods

We used transgene stacking to place a human CD55 transgene adjacent to a human heme oxygenase 1 construct at the porcine ROSA26 locus. A transgenic pig was analyzed by PCR, RT‐PCR, droplet digital PCR, immunohistochemistry, immunofluorescence, and flow cytometry. Resistance to complement‐mediated cell lysis and caspase 3/7 activation were determined in vitro.

Results

The ROSA26 locus was retargeted efficiently, and animals were generated by nuclear transfer. RNA and protein analyses revealed abundant expression in all organs analyzed, including pancreatic beta cells. Transgenic porcine kidney fibroblasts were almost completely protected against complement‐mediated lysis and showed reduced caspase 3/7 activation.

Conclusion

Step‐by‐step placement enables highly expressed single‐copy xenoprotective transgenes to be grouped at porcine ROSA26.
The role of hypoxia‐inducible factors in carotid body (patho) physiology
Authors: Semenza Gregg L. , Prabhakar Nanduri R.
Published: January 2018

Hypoxia‐inducible factors mediate adaptive responses to reduced O2 availability. In patients with obstructive sleep apnoea, repeated episodes of hypoxaemia and reoxygenation (intermittent hypoxia) are sensed by the carotid body (CB). The ensuing CB chemosensory reflex activates the sympathetic nervous system and increased secretion of catecholamines by the adrenal medulla, resulting in hypertension and breathing abnormalities. In the CB, intermittent hypoxia induces the formation of reactive oxygen species (ROS) and increased intracellular Ca2+ levels, which drive increased expression of hypoxia‐inducible factor (HIF) 1α and a decrease in the levels of HIF‐2α.  Intermittent hypoxia increases HIF‐1α‐dependent expression of Nox2, encoding the pro‐oxidant enzyme NADPH oxidase 2, and decreased HIF‐2α‐dependent expression of Sod2, encoding the anti‐oxidant enzyme superoxide dismutase 2. These changes in gene expression drive persistently elevated ROS levels in the CB, brainstem, and adrenal medulla that are required for the development of hypertension and breathing abnormalities. The ROS generated by dysregulated HIF activity in the CB results in oxidation and inhibition of haem oxygenase 2, and the resulting reduction in the levels of carbon monoxide leads to increased hydrogen sulfide production, triggering glomus cell depolarization. Thus, the pathophysiology of obstructive sleep apnoea involves the dysregulation of O2‐regulated transcription factors, gasotransmitters, and sympathetic outflow that affects blood pressure and breathing.

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Susceptibility of sweet and sour cherry cultivars/genotypes to feeding damage caused by Bryobia rubrioculus (Acari: Tetranychidae)
Authors: Honarparvar Nazila , Khanjani Mohammad , Zemek Rostislav , Bouzari Naser
Published: January 2018

The sweet cherry Prunus avium L. and sour cherry Prunus cerasus L. are considered important fruit trees, providing valuable and delicious fruit worldwide. We compared the susceptibility of 10 cultivars of sweet cherry and 5 genotypes of sour cherry to Bryobia rubrioculus Scheuten feeding by manually counting brown spots and by measuring damaged leaf area using a computerized image analysis technique. Damaged leaves were individually scanned on a common flatbed color scanner on a white background. The cultivars and genotypes of sweet and sour cherries, based on the mean area percentage having feeding symptoms on the leaves, were classified into three groups: (1) "susceptible", i.e., Hamedan, BN 5150, BT 5148, Zard-90, KB 9 and KB 10; (2) "semi-resistant", i.e., KB 21, Sabima, BT 5124, KB 25 and Lambert; and (3) "resistant", i.e., Haj Yousefi, BO 5187, BT 5154 and Siah Mashhad. A corresponding increase in the mean value of the symptom area was observed as the number of adult brown mites increased per disc. The results of both methods were compared, and the advantages of each method were discussed.
A new species of Neobonzia (Acariformes: Cunaxidae, Coleoscirinae) with a key to species of China
Authors: Chen Jian-Xin , Guo Jian-Jun , Yi Tian-Ci , Jin Dao-Chao
Published: January 2018

This paper presents a new species from China, Neobonzia neomalookensis sp. nov. which is described and illustrated in detail based on adult females and protonymphs. This new species resembles N. malookensis Bashir & Afzal, 2009, but can be easily distinguished by the following features: Palp tibiotarsus with 1 spine-like seta, 1 tubercle and 4 simple setae. Genu III with 1 asl and genu IV with 1 asl. Tibia I with 1 bsl, tarsus I with 3 bsl. A key to females of China is given.
Use of time-of-flight secondary ion mass spectrometry for the investigation of hydrogen-induced effects in austenitic steel AISI 304L
Authors: Röhsler Andreas , Sobol Oded , Nolze Gert , Unger Wolfgang E. S. , Böllinghaus Thomas
Published: January 2018

During the energy transformation from fossil fuels to renewable energy sources, the use of hydrogen as fuel and energy storage can play a key role. This presents new challenges to industry and the scientific community alike. The storage and transport of hydrogen, which is nowadays mainly realized by austenitic stainless steels, remains problematic [L. Zhang et al., Int. J. Hydrogen Energy 39, 20578 (2014)], which is due to the degradation of mechanical properties and the possibility of phase transformation by hydrogen diffusion and accumulation [P. Rozenak, Metall. Mater. Trans. A 45, 162 (2014)]. The development of materials and technologies requires a fundamental understanding of these degradation processes. Therefore, studying the behavior of hydrogen in austenitic steel contributes to an understanding of the damage processes, which is crucial for both life assessment and safe use of components in industry and transportation. As one of the few tools that is capable of depicting the distribution of hydrogen in steels, time-of-flight secondary ion mass spectrometry was conducted after electrochemical charging [O. Sobol et al., Surf. Interface Anal. 48, 474 (2016)]. To obtain further information about the structural composition and cracking behavior, electron-backscattered diffraction and scanning electron microscopy were performed. Gathered data of chemical composition and topography were treated employing data fusion, thus creating a comprehensive portrait of hydrogen-induced effects in the austenite grade AISI 304L. Specimens were electrochemically charged with deuterium instead of hydrogen. This arises from the difficulties to distinguish between artificially charged hydrogen and traces existing in the material or the rest gas in the analysis chamber. Similar diffusion and permeation behavior, as well as solubility, allow nonetheless to draw conclusions from the experiments [Y. Fukai and H. Sugimoto, Adv. Phys. 34, 263 (1985)].
A constitutive model for simple shear of dense frictional suspensions
Authors: Singh Abhinendra , Mari Romain , Denn Morton M. , Morris Jeffrey F.
Published: January 2018

Discrete particle simulations are used to study the shear rheology of dense, stabilized, frictional particulate suspensions in a viscous liquid, toward development of a constitutive model for steady shear flows at arbitrary stress. These suspensions undergo increasingly strong continuous shear thickening (CST) as the solid volume fraction [Formula: see text] increases above a critical volume fraction, and discontinuous shear thickening (DST) is observed in a range of [Formula: see text]. When studied at controlled stress, the DST behavior is associated with nonmonotonic flow curves of the steady-state shear rate as a function of stress. Recent studies have related shear thickening to a transition from mostly lubricated to predominantly frictional contacts with the increase in stress. In this study, the behavior is simulated over wide ranges of concentration, dimensionless shear stress, and coefficient of interparticle friction. The simulation data have been used to populate the lubricated-to-frictional rheology model of Wyart and Cates [Phys. Rev. Lett. 112, 098302 (2014)], which is based on the concept of two viscosity divergences or “jamming” points at volume fraction [Formula: see text] (random close packing) for the low-stress lubricated state, and at [Formula: see text] for any nonzero μ in the frictional state; a generalization provides the normal stress response as well as the shear stress, allowing a description of the full stress tensor. A flow state map of this material is developed based on the simulation results. At low stress and/or intermediate [Formula: see text], the system exhibits CST, and DST appear at volume fractions below but approaching the frictional jamming point. For [Formula: see text], DST is associated with a material transition from one stress-independent rheology to another, while for [Formula: see text], the system exhibits DST to shear jamming as the stress increases.
Characterization of the
In Vitro
Chlamydia pecorum Response to Gamma Interferon
Authors: Islam M. Mominul , Jelocnik Martina , Huston Wilhelmina M. , Timms Peter , Polkinghorne Adam
Published: January 2018

Chlamydia pecorum
is an important intracellular bacterium that causes a range of diseases in animals, including a native Australian marsupial, the koala. In humans and animals, a gamma interferon (IFN-γ)-mediated immune response is important for the control of intracellular bacteria. The present study tested the hypotheses that
C. pecorum
can escape IFN-γ-mediated depletion of host cell tryptophan pools. In doing so, we demonstrated that, unlike
Chlamydia trachomatis
,
C. pecorum
is completely resistant to IFN-γ in human epithelial cells. While the growth of
C. pecorum
was inhibited in tryptophan-deficient medium, it could be restored by the addition of kynurenine, anthranilic acid, and indole, metabolites that could be exploited by the gene products of the
C. pecorum
tryptophan biosynthesis operon. We also found that expression of
trp
genes was detectable only when
C. pecorum
was grown in tryptophan-free medium, with gene repression occurring in response to the addition of kynurenine, anthranilic acid, and indole. When grown in bovine kidney epithelial cells, bovine IFN-γ also failed to restrict the growth of
C. pecorum
, while
C. trachomatis
was inhibited, suggesting that
C. pecorum
could use the same mechanisms to evade the immune response
in vivo
in its natural host. Highlighting the different mechanisms triggered by IFN-γ, however, both species failed to grow in murine McCoy cells treated with murine IFN-γ. This work confirms previous hypotheses about the potential survival of
C. pecorum
after IFN-γ-mediated host cell tryptophan depletion and raises questions about the immune pathways used by the natural hosts of
C. pecorum
to control the widespread pathogen.
Purification of Highly Active Alphavirus Replication Complexes Demonstrates Altered Fractionation of Multiple Cellular Membranes
Authors: Pietilä Maija K. , van Hemert Martijn J. , Ahola Tero
Published: January 2018

Positive-strand RNA viruses replicate their genomes in membrane-associated structures; alphaviruses and many other groups induce membrane invaginations called spherules. Here, we established a protocol to purify these membranous replication complexes (RCs) from cells infected with Semliki Forest virus (SFV). We isolated SFV spherules located on the plasma membrane and further purified them using two consecutive density gradients. This revealed that SFV infection strongly modifies cellular membranes. We removed soluble proteins, the Golgi membranes, and most of the mitochondria, but plasma membrane, endoplasmic reticulum (ER), and late endosome markers were retained in the membrane fraction that contained viral RNA synthesizing activity, replicase proteins, and minus- and plus-strand RNA. Electron microscopy revealed that the purified membranes displayed spherule-like structures with a narrow neck. This membrane enrichment was specific to viral replication, as such a distribution of membrane markers was only observed after infection. Besides the plasma membrane, SFV infection remodeled the ER, and the cofractionation of the RC-carrying plasma membrane and ER suggests that SFV recruits ER proteins or membrane to the site of replication. The purified RCs were highly active in synthesizing both genomic and subgenomic RNA. Detergent solubilization destroyed the replication activity, demonstrating that the membrane association of the complex is essential. Most of the newly made RNA was in double-stranded replicative molecules, but the purified complexes also produced single-stranded RNA as well as released newly made RNA. This indicates that the purification established here maintained the functionality of RCs and thus enables further structural and functional studies of active RCs.

IMPORTANCE
Similar to all positive-strand RNA viruses, the arthropod-borne alphaviruses induce membranous genome factories, but little is known about the arrangement of viral replicase proteins and the presence of host proteins in these replication complexes. To improve our knowledge of alphavirus RNA-synthesizing complexes, we isolated and purified them from infected mammalian cells. Detection of viral RNA and
in vitro
replication assays revealed that these complexes are abundant and highly active when located on the plasma membrane. After multiple purification steps, they remain functional in synthesizing and releasing viral RNA. Besides the plasma membrane, markers for the endoplasmic reticulum and late endosomes were enriched with the replication complexes, demonstrating that alphavirus infection modified cellular membranes beyond inducing replication spherules on the plasma membrane. We have developed here a gentle purification method to obtain large quantities of highly active replication complexes, and similar methods can be applied to other positive-strand RNA viruses.
Retinal vascular alterations in reticular pseudodrusen with and without outer retinal atrophy assessed by optical coherence tomography angiography
Authors: Cicinelli Maria Vittoria , Rabiolo Alessandro , Sacconi Riccardo , Lamanna Francesca , Querques Lea , Bandello Francesco , Querques Giuseppe
Published: January 2018

Purpose

To investigate the intraretinal structural and vascular alterations in patients featuring reticular pseudodrusen (RPD), RPD with outer retinal atrophy (ORA), and drusen.

Design

Observational cross-sectional study.

Methods

Clinical practice study including 68 eyes of 57 patients (22 eyes of 17 patients with RPD; 24 eyes of 21 patients with RPD+ORA; 22 eyes of 19 patients with drusen). Each patient underwent spectral-domain optical coherence tomography (OCT) and OCT angiography (OCT-A). Measurement of retinal layers’ thickness was obtained by the automated segmentation protocol of the Spectralis OCT (Heidelberg Eye Explorer V.1.9.10.0). The superficial capillary plexus (SCP) and the deep capillary plexus (DCP) vessel density, as well as the size of the foveal avascular zone were calculated on 3×3 OCT-A. Main outcome was to compare vessel density at the SCP and DCP among the groups and controls.

Results

At the SCP, the vessel density was lower in RPD and RPD+ORA patients with respect to controls (P=0.02 and P=0.003, respectively). At the DCP, meaningful disparity was found between the study groups and the healthy subjects in the vessel density (P<0.001, P=0.04 and P=0.001 for RPD, RDP+ORA and drusen, respectively). The ganglion cell layer (GCL) was thinner in all patients affected either by RPD, RPD+ORA or drusen compared with healthy subjects (P=0.02, P=0.03 and P=0.004, respectively).

Conclusion

Significant retinal vascular loss is a common feature of patients with non-exudative age-related macular degeneration, more pronounced in those featuring RPD and RPD+ORA. It is associated with retinal thinning, localised particularly at the GCL, compared with controls.
Cell of origin affects tumour development and phenotype in pancreatic ductal adenocarcinoma
Authors: Lee Alex Y L , Dubois Claire L , Sarai Karnjit , Zarei Soheila , Schaeffer David F , Sander Maike , Kopp Janel L
Published: January 2018

Objective

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive tumour thought to arise from ductal cells via pancreatic intraepithelial neoplasia (PanIN) precursor lesions. Modelling of different genetic events in mice suggests both ductal and acinar cells can give rise to PDAC. However, the impact of cellular context alone on tumour development and phenotype is unknown.

Design

We examined the contribution of cellular origin to PDAC development by inducing PDAC-associated mutations, KrasG12D expression and Trp53 loss, specifically in ductal cells (Sox9CreER;KrasLSL-G12D;Trp53flox/flox (‘Duct:KPcKO’)) or acinar cells (Ptf1aCreER;KrasLSL-G12D;Trp53flox/flox (‘Acinar:KPcKO’)) in mice. We then performed a thorough analysis of the resulting histopathological changes.

Results

Both mouse models developed PDAC, but Duct:KPcKO mice developed PDAC earlier than Acinar:KPcKO mice. Tumour development was more rapid and associated with high-grade murine PanIN (mPanIN) lesions in Duct:KPcKO mice. In contrast, Acinar:KPcKO mice exhibited widespread metaplasia and low-grade as well as high-grade mPanINs with delayed progression to PDAC. Acinar-cell-derived tumours also had a higher prevalence of mucinous glandular features reminiscent of early mPanIN lesions.

Conclusion

These findings indicate that ductal cells are primed to form carcinoma in situ that become invasive PDAC in the presence of oncogenic Kras and Trp53 deletion, while acinar cells with the same mutations appear to require a prolonged period of transition or reprogramming to initiate PDAC. Our findings illustrate that PDAC can develop in multiple ways and the cellular context in which mutations are acquired has significant impact on precursor lesion initiation, disease progression and tumour phenotype.
Consent and the ethical duty to participate in health data research
Authors: Ballantyne Angela , Schaefer G Owen
Published: January 2018

The predominant view is that a study using health data is observational research and should require individual consent unless it can be shown that gaining consent is impractical. But recent arguments have been made that citizens have an ethical obligation to share their health information for research purposes. In our view, this obligation is sufficient ground to expand the circumstances where secondary use research with identifiable health information is permitted without explicit subject consent. As such, for some studies the Institutional Review Board/Research Ethics Committee review process should not assess the practicality of gaining consent for data use. Instead the review process should focus on assessing the public good of the research, public engagement and transparency.