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1. Independent Lec1A CHO Glycosylation Mutants Arise from Point Mutations in N-Acetylglucosaminyltransferase I That Reduce Affinity for Both Substrates. Molecular Consequences Based on the Crystal Structure of GlcNAc-TI

Author

Wei Chen, Ulug M. Ünligil, James M. Rini, and Pamela Stanley

Subjects
Glycosylation, Transcription, Genetic, Arginine, Molecular Sequence Data, Mutant, CHO Cells, Biology, Crystallography, X-Ray, N-Acetylglucosaminyltransferases, Biochemistry, Substrate Specificity, chemistry.chemical_compound, Cricetulus, Cricetinae, Aspartic acid, Animals, Point Mutation, Asparagine, Conserved Sequence, chemistry.chemical_classification, Aspartic Acid, Chinese hamster ovary cell, Point mutation, Amino acid, carbohydrates (lipids), Phenotype, chemistry, Mutagenesis, Site-Directed, Rabbits, Protein Binding
Abstract

A key enzyme in regulating the maturation of N-linked glycans is UDP-N-acetylglucosamine:alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I (GlcNAc-TI, EC 2.4.1.101). Lec1 CHO cells lack GlcNAc-TI activity and synthesize only the oligomannosyl class of N-glycans. By contrast, Lec1A CHO mutants have weak GlcNAc-TI activity due to the reduced affinity of GlcNAc-TI for both the UDP-GlcNAc and Man(5)GlcNAc(2)Asn substrates. Lec1A CHO mutants synthesize hybrid and complex N-glycans, albeit in reduced amounts compared to parental CHO cells. In this paper, we identify two point mutations that gave rise to the Lec1A phenotype in three independent Lec1A CHO mutants. The G634A mutation in Lec1A.2C converts an aspartic acid to an asparagine at amino acid 212, disrupting a conserved DXD motif (E(211)DD(213) in all GlcNAc-TIs) that makes critical interactions with bound UDP-GlcNAc and Mn(2+) ion in rabbit GlcNAc-TI. The C907T mutation in Lec1A.3E and Lec1A.5J converts an arginine conserved in all GlcNAc-TIs to a tryptophan at amino acid 303, altering interactions that are important in stabilizing a critical structural element in rabbit GlcNAc-TI. Correction of each mutation by site-directed mutagenesis restored their GlcNAc-TI activity and lectin binding properties to parental levels. The effect of the two amino acid changes on GlcNAc-TI catalysis is discussed in relation to the crystal structure of rabbit GlcNAc-TI complexed with manganese and UDP-GlcNAc.

Published
2001

2. Treatment of ventilator-associated pneumonia (VAP) caused by Acinetobacter: results of prospective and multicenter ID-IRI study

Author

Jordi Rello, Alper Şener, Meliha Meric-Koc, Yasemin Cag, Mustafa Dogan, Fazilet Duygu, Esmeray Mutlu-Yilmaz, Tumer Guven, Rodrigo Hasbun, Selma Güler, Hakan Erdem, Serkan Oncu, Yasemin Akkoyunlu, Arzu Dogru, Ozgur Dagli, Zuhal Karakurt, Serhat Uysal, Serap Gencer, Oguz Karabay, Hale Turan, Güven Çelebi, Gül Durmuş, Rezan Harman, Ayse Batirel, Mehmet Ulug, Asuman Inan, Emel Aslan, Yesim Uygun, Selma Tosun, Gülden Ersöz, Erdem, H, Cag, Y, Gencer, S, Uysal, S, Karakurt, Z, Harman, R, Aslan, E, Mutlu-Yilmaz, E, Karabay, O, Uygun, Y, Ulug, M, Tosun, S, Dogru, A, Sener, A, Dogan, M, Hasbun, R, Durmus, G, Turan, H, Batirel, A, Duygu, F, Inan, A, Akkoyunlu, Y, Celebi, G, Ersoz, G, Guven, T, Dagli, O, Guler, S, Meric-Koc, M, Oncu, S, Rello, J, Sakarya Üniversitesi/Tıp Fakültesi/Dahili Tıp Bilimleri Bölümü, Karabay, Oğuz, AKKOYUNLU, YASEMİN, and Zonguldak Bülent Ecevit Üniversitesi

Subjects
0301 basic medicine, Male, Intensive-Care Units, Antibiotics, Resistance, results of prospective and multicenter ID-IRI study.-, European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology, 2019 [Erdem H., Cag Y., Gencer S., Uysal S., Karakurt Z., Harman R., Aslan E., Mutlu-Yilmaz E., Karabay O., Uygun Y., et al., -Treatment of ventilator-associated pneumonia (VAP) caused by Acinetobacter], law.invention, chemistry.chemical_compound, 0302 clinical medicine, Medical microbiology, law, Risk Factors, 030212 general & internal medicine, Prospective Studies, Lung, biology, Acinetobacter, Ventilator-associated pneumonia, Pneumonia, Ventilator-Associated, General Medicine, Middle Aged, Antimicrobial, Baumannii, Intensive care unit, Management, Anti-Bacterial Agents, Infectious-Diseases Society, Intensive Care Units, Infectious Diseases, Female, Original Article, Acinetobacter Infections, Microbiology (medical), medicine.medical_specialty, medicine.drug_class, 030106 microbiology, America, Guidelines, Microbiology, 03 medical and health sciences, Internal medicine, medicine, Humans, Mortality, Aged, Creatinine, business.industry, Pneumonia, medicine.disease, biology.organism_classification, Treatment, chemistry, VAP, business
Abstract

Ventilator-associated pneumonia (VAP) due to Acinetobacter spp. is one of the most common infections in the intensive care unit. Hence, we performed this prospective-observational multicenter study, and described the course and outcome of the disease. This study was performed in 24 centers between January 06, 2014, and December 02, 2016. The patients were evaluated at time of pneumonia diagnosis, when culture results were available, and at 72 h, at the 7th day, and finally at the 28th day of follow-up. Patients with coexistent infections were excluded and only those with a first VAP episode were enrolled. Logistic regression analysis was performed. A total of 177 patients were included; empiric antimicrobial therapy was appropriate (when the patient received at least one antibiotic that the infecting strain was ultimately shown to be susceptible) in only 69 (39%) patients. During the 28-day period, antibiotics were modified for side effects in 27 (15.2%) patients and renal dose adjustment was made in 38 (21.5%). Ultimately, 89 (50.3%) patients died. Predictors of mortality were creatinine level (OR, 1.84 (95% CI 1.279–2.657); p = 0.001), fever (OR, 0.663 (95% CI 0.454–0.967); p = 0.033), malignancy (OR, 7.095 (95% CI 2.142–23.500); p = 0.001), congestive heart failure (OR, 2.341 (95% CI 1.046–5.239); p = 0.038), appropriate empiric antimicrobial treatment (OR, 0.445 (95% CI 0.216–0.914); p = 0.027), and surgery in the last month (OR, 0.137 (95% CI 0.037–0.499); p = 0.003). Appropriate empiric antimicrobial treatment in VAP due to Acinetobacter spp. was associated with survival while renal injury and comorbid conditions increased mortality. Hence, early diagnosis and appropriate antibiotic therapy remain crucial to improve outcomes. © 2019, Springer-Verlag GmbH Germany, part of Springer Nature.

Published
2019

3. Comparison of brucellar and tuberculous spondylodiscitis patients: results of the multicenter 'Backbone-1 Study'

Author

Tumer Guven, Hakan Erdem, Rehab H. El-Sokkary, Dilara Inan, Serda Gulsun, Fatma Yilmaz-Karadag, Gunay Tuncer-Ertem, Elif Sahin Horasan, Şafak Kaya, Ali İrfan Baran, Recep Tekin, Archontoula Fragou, Seniha Senbayrak, Esra Arslanalp, Fatma Sirmatel, Selçuk Kaya, Ayşe Willke, Salih Cesur, Sani H. Aliyu, Ergys Ramosaco, Meliha Meric-Koc, Meltem Taşbakan, Sukran Kose, Gönül Aslan, Efthymia Giannitsioti, Gonul Sengoz, Haluk Vahaboglu, Nazif Elaldi, Selma Ates-Guler, Birgul Mete, Sesin Kocagöz, Hulya Kusoglu, Ertugrul Guclu, Dilek Yildiz Sevgi, Hava Yilmaz, Gulfem Akengin Ocal, Ayse Batirel, Behiye Dede, Birsen Cetin, Tunc Oktenoglu, D. Ashraf ALGallad, Mehmet Ulug, Sebnem Senol, Banu Karaca, Ilker Inanc Balkan, Aygul Dogan-Celik, Nural Bekiroglu, Gonul Cicek-Senturk, Filiz Pehlivanoglu, Asuman Inan, MERİÇ KOÇ, MELİHA, Erdem, H, Elaldi, N, Batirel, A, Aliyu, S, Sengoz, G, Pehlivanoglu, F, Ramosaco, E, Gulsun, S, Tekin, R, Mete, B, Balkan, II, Sevgi, DY, Giannitsioti, E, Fragou, A, Kaya, S, Cetin, B, Oktenoglu, T, DoganCelik, A, Karaca, B, Horasan, ES, Ulug, M, Man, A, Arslanalp, E, Ates-Guler, S, Willke, A, Senol, S, Inan, D, Guclu, E, Tuncer-Ertem, G, Meric-Koc, M, Tasbakan, M, Senbayrak, S, Cicek-Senturk, G, Sirmatel, F, Ocal, G, Kocagoz, S, Kusoglu, H, Guven, T, Baran, AI, Dede, B, Yilmaz-Karadag, F, Kose, S, Yilmaz, H, Asian, G, Algallad, DA, Cesur, S, El-Sokkary, R, Bekiroglu, N, Vahaboglu, H, Sakarya Üniversitesi/Tıp Fakültesi/Dahili Tıp Bilimleri Bölümü, Güçlü, Ertuğrul, [Erdem, Hakan] Culhane Med Acad, Dept Infect Dis & Clin Microbiol, Ankara, Turkey -- [Elaldi, Nazif] Cumhuriyet Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Sivas, Turkey -- [Batirel, Ayse -- Ocal, Gulfem] Dr Lao Kirdar Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Aliyu, Sani] Cambridge Univ Hosp Fdn Trust, Addenbrookes Hosp, Clin Microbiol & Publ Hlth Lab, Cambridge, England -- [Sengoz, Gonul -- Pehlivanoglu, Filiz] Haseki Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Ramosaco, Ergys] Univ Hosp Ctr Mother Teresa, Infect Dis Hosp, Tirana, Albania -- [Gulsun, Serda -- Kaya, Safak] Diyarbakir Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Diyarbakir, Turkey -- [Tekin, Recep] Dicle Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Diyarbakir, Turkey -- [Mete, Birgul -- Balkan, Ilker Inanc] Istanbul Univ, Cerrahpasa Sch Med, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Sevgi, Dilek Yildiz] Sisli Etfal Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Giannitsioti, Efthymia -- Fragou, Archontoula] Attikon Univ Gen Hosp, Athens Univ Med Sch, Dept Internal Med, Athens, Greece -- [Kaya, Selcuk] Karadeniz Tech Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Trabzon, Turkey -- [Cetin, Birsen] Koc Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Oktenoglu, Tune] Koc Univ, Sch Med, Dept Neurosurg, Istanbul, Turkey -- [DoganCelik, Aygul] Trakya Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Edirne, Turkey -- [Karaca, Banu] Izmir Bozyaka Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Izmir, Turkey -- [Horasan, Elif Sahin] Mersin Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Mersin, Turkey -- [Ulug, Mehmet] Private Umit Hosp, Dept Infect Dis & Clin Microbiol, Eskisehir, Turkey -- [Man, Asuman -- Senbayrak, Seniha] Haydarpasa Numune Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Arslanalp, Esra -- Willke, Ayse -- Meric-Koc, Meliha] Kocaeli Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Kocaeli, Turkey -- [Ates-Guler, Selma] Kahramanmaras Sutcu Imam Univ, Sch Med, Dept Infect Dis & Clin Microbiol, TR-46050 Kahramanmaras, Turkey -- [Senol, Sebnem] Celal Bayar Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Manisa, Turkey -- [Inan, Dilara] Akdeniz Univ, Sch Med, Dept Infect Dis & Clin Microbiol, TR-07058 Antalya, Turkey -- [Guclu, Ertugrul] Sakarya Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Sakarya, Turkey -- [Tuncer-Ertem, Gunay -- Cesur, Salih] Ankara Numune Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Ankara, Turkey -- [Tasbakan, Meitem] Ege Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Izmir, Turkey -- [Cicek-Senturk, Gonul -- Asian, Gonul] Diskapi Yildirim Beyazit Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Ankara, Turkey -- [Sirmatel, Fatma] Abant Izzet Baysal Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Bolu, Turkey -- [Kocagoz, Sesin -- Kusoglu, Hulya] Acibadem Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Guven, Turner] Ankara Ataturk Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Ankara, Turkey -- [Baran, Ali Irfan] Yuzuncu Yil Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Van, Turkey -- [Dede, Behiye] Umraniye Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Yilmaz-Karadag, Fatma -- Vahaboglu, Haluk] Medeniyet Univ, Goztepe Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Istanbul, Turkey -- [Kose, Sukran] Tepecik Training & Res Hosp, Dept Infect Dis & Clin Microbiol, Izmir, Turkey -- [Yilmaz, Hava] Ondokuz Mayis Univ, Sch Med, Dept Infect Dis & Clin Microbiol, Samsun, Turkey -- [Algallad, D. Ashraf -- El-Sokkary, Rehab] Zagazig Univ Hosp, Infect Control Unit, Az Zagazig, Egypt -- [Bekiroglu, Nural] Marmara Univ, Sch Med, Dept Biostat, Istanbul, Turkey, VAHABOGLU, Haluk -- 0000-0001-8217-1767, and Ondokuz Mayıs Üniversitesi

Subjects
Adult, Male, Spondylodiscitis, endocrine system, medicine.medical_specialty, Discitis, Constitutional symptoms, Context (language use), Brucellosis, Sequelae, Internal medicine, medicine, Back pain, Humans, Tuberculosis, Orthopedics and Sports Medicine, Abscess, Outcome, Aged, medicine.diagnostic_test, business.industry, Middle Aged, medicine.disease, Surgery, Orthopedics, Erythrocyte sedimentation rate, Female, Neurology (clinical), medicine.symptom, Complication, business
Abstract

WOS: 000366655100045, PubMed ID: 26386176, BACKGROUND CONTEXT: No direct comparison between brucellar spondylodiscitis (BSD) and tuberculous spondylodiscitis (TSD) exists in the literature. PURPOSE: This study aimed to compare directly the clinical features, laboratory and radiological aspects, treatment, and outcome data of patients diagnosed as BSD and TSD. STUDY DESIGN: A retrospective, multinational, and multicenter study was used. PATIENT SAMPLE: A total of 641 (TSD, 314 and BSD, 327) spondylodiscitis patients from 35 different centers in four countries (Turkey, Egypt, Albania, and Greece) were included. OUTCOME MEASURES: The pre- and peri- or post-treatment spinal deformity and neurologic deficit parameters, and mortality were carried out. METHODS: Brucellar spondylodiscitis and TSD groups were compared for demographics, clinical, laboratory, radiological, surgical interventions, treatment, and outcome data. The Student t test and Mann-Whitney U test were used for group comparisons. Significance was analyzed as two sided and inferred at 0.05 levels. RESULTS: The median baseline laboratory parameters including white blood cell count, C-reactive protein, and erythrocyte sedimentation rate were higher in TSD than BSD (p

Published
2015

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