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1.
Biochemistry ; 63(13): 1599-1607, 2024 Jul 02.
Article in English | MEDLINE | ID: mdl-38907702

ABSTRACT

Small-scale bioreactors that are affordable and accessible would be of major benefit to the research community. In previous work, an open-source, automated bioreactor system was designed to operate up to the 30 mL scale with online optical monitoring, stirring, and temperature control, and this system, dubbed Chi.Bio, is now commercially available at a cost that is typically 1-2 orders of magnitude less than commercial bioreactors. In this work, we further expand the capabilities of the Chi.Bio system by enabling continuous pH monitoring and control through hardware and software modifications. For hardware modifications, we sourced low-cost, commercial pH circuits and made straightforward modifications to the Chi.Bio head plate to enable continuous pH monitoring. For software integration, we introduced closed-loop feedback control of the pH measured inside the Chi.Bio reactors and integrated a pH-control module into the existing Chi.Bio user interface. We demonstrated the utility of pH control through the small-scale depolymerization of the synthetic polyester, poly(ethylene terephthalate) (PET), using a benchmark cutinase enzyme, and compared this to 250 mL bioreactor hydrolysis reactions. The results in terms of PET conversion and rate, measured both by base addition and product release profiles, are statistically equivalent, with the Chi.Bio system allowing for a 20-fold reduction of purified enzyme required relative to the 250 mL bioreactor setup. Through inexpensive modifications, the ability to conduct pH control in Chi.Bio reactors widens the potential slate of biochemical reactions and biological cultivations for study in this system, and may also be adapted for use in other bioreactor platforms.


Subject(s)
Bioreactors , Polyethylene Terephthalates , Polyethylene Terephthalates/chemistry , Polyethylene Terephthalates/metabolism , Hydrogen-Ion Concentration , Hydrolysis , Carboxylic Ester Hydrolases/metabolism , Carboxylic Ester Hydrolases/chemistry , Burkholderiales/enzymology , Burkholderiales/metabolism , Software
2.
Sci Rep ; 14(1): 14449, 2024 06 24.
Article in English | MEDLINE | ID: mdl-38914665

ABSTRACT

As genomic databases expand and artificial intelligence tools advance, there is a growing demand for efficient characterization of large numbers of proteins. To this end, here we describe a generalizable pipeline for high-throughput protein purification using small-scale expression in E. coli and an affordable liquid-handling robot. This low-cost platform enables the purification of 96 proteins in parallel with minimal waste and is scalable for processing hundreds of proteins weekly per user. We demonstrate the performance of this method with the expression and purification of the leading poly(ethylene terephthalate) hydrolases reported in the literature. Replicate experiments demonstrated reproducibility and enzyme purity and yields (up to 400 µg) sufficient for comprehensive analyses of both thermostability and activity, generating a standardized benchmark dataset for comparing these plastic-degrading enzymes. The cost-effectiveness and ease of implementation of this platform render it broadly applicable to diverse protein characterization challenges in the biological sciences.


Subject(s)
Escherichia coli , Robotics , Robotics/methods , Escherichia coli/genetics , Protein Engineering/methods , High-Throughput Screening Assays/methods , High-Throughput Screening Assays/economics , Hydrolases/metabolism , Hydrolases/chemistry , Hydrolases/genetics , Polyethylene Terephthalates/chemistry , Reproducibility of Results
3.
Ocul Immunol Inflamm ; 31(1): 142-148, 2023 Jan.
Article in English | MEDLINE | ID: mdl-34797735

ABSTRACT

BACKGROUND: In accordance with worldwide data, the Robert Koch Institute (RKI) has reported a constant increase of syphilis cases in Germany over the past decade. METHODS: We analysed the data of all patients, referred to a Department of Ophthalmology in a tertiary referral centre in Düsseldorf, Germany between 2008 and 2019, who were tested for syphilis. The epidemiologic, demographic, clinical, diagnostic and therapeutic data were retrieved from the records and evaluated in a retrospective, descriptive, non-comparative study. RESULTS: Syphilis serology was positive in 32/1840 (1.7%) patients, and was evenly distributed over this period. 26 (81.3%) were male, 19 (59.4%) belonged to a risk group. Ocular syphilis was the primary diagnosis for 29 patients (90.6%). The most frequent manifestation was uveitis (n = 20, 62.5%). By the end of therapy, 19 patients (59.4%) had an improved visual acuity. CONCLUSION: The incidence of ocular syphilis cases has remained stable over the last decade.


Subject(s)
Eye Infections, Bacterial , Syphilis , Tertiary Care Centers , Female , Humans , Male , Eye Infections, Bacterial/diagnosis , Eye Infections, Bacterial/epidemiology , Germany/epidemiology , Ophthalmology , Retrospective Studies , Syphilis/diagnosis , Syphilis/epidemiology , Syphilis Serodiagnosis , Tertiary Care Centers/statistics & numerical data , Uveitis/epidemiology , Referral and Consultation
4.
Curr Eye Res ; 45(12): 1484-1489, 2020 12.
Article in English | MEDLINE | ID: mdl-32434387

ABSTRACT

BACKGROUND AND PURPOSE: In vivo confocal microscopy (IVCM) is a non-invasive imaging technique that allows morphological analysis as a diagnostic approach of the cornea in real time, thus providing a suspected diagnosis of fungal or amoebic keratitis immediately, whereas culture or PCR require several days or even weeks. Since these infections are rare, it is difficult for ophthalmologists to gain the experience necessary to differentiate infection from normal findings or artefacts. The purpose of this project was to establish a simulator, on which physicians could practice as well as acquiring a database of IVCM images of fungal or amoebic keratitis and respective analyses. PATIENTS AND METHODS: An IVCM simulator was set up with cadaver human corneas, infected with either acanthamoeba, candida or aspergillus. Twenty-one ophthalmologists were trained in IVC microscopy first in a Dry Lab, then practically on the simulator. For evaluation, the participants were asked to fill out a standardized questionnaire, with a pre- and post-course self-assessment. RESULTS: The self-assessed theoretical and practical skills in differentiating infectious from non-infectious keratitis in IVCM significantly increased (p = 0.0001, p = 0.0002, respectively). The barrier to use this technique decreased (p = 0.0474). CONCLUSION: A very simple protocol based on a model of ex vivo corneal mycotic and amoebic infections can be used to train novices in the structured approach and diagnostic use of IVCM for corneal infections.


Subject(s)
Acanthamoeba Keratitis/diagnosis , Aspergillosis/diagnosis , Candidiasis/diagnosis , Corneal Ulcer/diagnosis , Eye Infections, Fungal/diagnosis , Microscopy, Confocal/instrumentation , Simulation Training/methods , Aspergillosis/microbiology , Candidiasis/microbiology , Corneal Ulcer/microbiology , Equipment Design , Eye Infections, Fungal/microbiology , Female , Humans , Male , Surveys and Questionnaires
5.
Ophthalmologe ; 116(10): 957-966, 2019 Oct.
Article in German | MEDLINE | ID: mdl-30810837

ABSTRACT

BACKGROUND AND PURPOSE: Mycotic keratitis is a serious but relatively rare disease. No targeted data collection in Germany existed until the foundation of the German Pilz-Keratitis Register in 2015. PATIENTS AND METHODS: The inclusion of retrospective and prospective patients was carried out. INCLUSION CRITERIA: diagnosis confirmed by the polymerase chain reaction (PCR), culture, histology or confocal microscopy (IVCM). Collected parameters: date of symptom onset, date and method of diagnosis, risk factors, visual acuity and findings at admission and at follow-up, conservative and surgical treatment. RESULTS: By January 2018, a total of 102 eyes from the years 2000-2017 were reported from 16 centers (64.3% female, mean age 52 years, range 18-95 years). The initial diagnosis was made correctly in only 20.6% of cases. The mean time to correct diagnosis was 31.7 ±â€¯46.9 (0-296) days. The diagnosis was confirmed in cultures in 74.5%, histologically in 30.4%, by PCR in 38.2% and IVCM in 27.4%. Fungal species identified were: 36.7% Fusarium spp., 35.8% Candida spp., 6.4% Aspergillus spp. and 21.1% other. The most important risk factor was the use of contact lenses. The most commonly used antifungal agent was voriconazole (64.7%) followed by amphotericin B (37.2%). Penetrating keratoplasty was performed in 65.7% of the cases and 8.8% of the affected eyes had to be enucleated. The visual acuity of the entire study population increased from the initial 0.16 ±â€¯0.25 (0.001-1.0) decimal to 0.28 ±â€¯0.34 (0-1.0) decimal. CONCLUSION: The correct diagnosis of fungal keratitis is often significantly delayed. The treatment can be very difficult and keratoplasty is often necessary. In order to gain a better understanding of this disease, to recognize previously unknown risk factors and, if necessary, a change in the spectrum of pathogens and to identify approaches to treatment optimization, the fungal keratitis registry will be continued.


Subject(s)
Eye Infections, Fungal , Adolescent , Adult , Aged , Aged, 80 and over , Antifungal Agents , Female , Germany , Humans , Male , Middle Aged , Prospective Studies , Registries , Retrospective Studies , Surveys and Questionnaires , Young Adult
6.
J Antimicrob Chemother ; 73(8): 2047-2053, 2018 08 01.
Article in English | MEDLINE | ID: mdl-29684150

ABSTRACT

Objectives: Aspergillus fumigatus is the most prevalent filamentous fungus in the respiratory tract of patients with cystic fibrosis (CF). The aim of this prospective multicentre study was to investigate the prevalence of azole-resistant A. fumigatus (ARAF) in respiratory secretions from CF patients across Germany and to characterize ARAF isolates by phenotypic and molecular methods. Methods: Twelve tertiary care centres from Germany participated in the study. In total, 2888 A. fumigatus isolates from 961 CF patients were screened for ARAF by using azole-containing agar plates. Antifungal susceptibility testing of isolates was performed by broth microdilution according to EUCAST guidelines. Analysis of mutations mediating resistance was performed using PCR and sequencing of the cyp51A gene. Furthermore, genotyping by microsatellite PCR was performed. Results: Of a total of 2888 A. fumigatus isolates, 101 isolates from 51 CF patients were found to be azole resistant (prevalence per patient 5.3%). The Essen centre had the highest prevalence (9.1%) followed by Munich (7.8%), Münster (6.0%) and Hannover (5.2%). Most ARAF isolates (n = 89) carried the TR34/L98H mutation followed by eight G54E/R, one TR46/Y121F/T289A and one F219S mutation. In two isolates no mutation was found. Genotyping results showed no major clustering. Forty-five percent of CF patients with ARAF had previously received azole therapy. Conclusions: This is the first multicentre study analysing the prevalence of ARAF isolates in German CF patients. Because of a resistance rate of up to 9%, susceptibility testing of A. fumigatus isolates from CF patients receiving antifungal treatment should be part of standard diagnostic work-up.


Subject(s)
Antifungal Agents/pharmacology , Aspergillus fumigatus/drug effects , Azoles/pharmacology , Cystic Fibrosis/microbiology , Drug Resistance, Fungal , Adult , Aspergillus fumigatus/genetics , Aspergillus fumigatus/isolation & purification , Cytochrome P-450 Enzyme System/genetics , DNA Mutational Analysis , Female , Fungal Proteins/genetics , Genotype , Germany , Humans , Male , Microbial Sensitivity Tests , Microsatellite Repeats , Mycological Typing Techniques , Prevalence , Prospective Studies
7.
Klin Monbl Augenheilkd ; 234(1): 40-45, 2017 Jan.
Article in German | MEDLINE | ID: mdl-28135744

ABSTRACT

Peri-ocular necrotising fasciitis is a very rare ophthalmological clinical picture and is potentially fatal. This disease is caused by bacterial infection of the fasciae, which rapidly spreads. The present article reports a typical case of the disease and gives an overview of the typical clinical signs and symptoms of peri-ocular necrotising fasciitis.


Subject(s)
Blepharitis/therapy , Eye Infections, Bacterial/therapy , Fasciitis, Necrotizing/therapy , Orbital Diseases/therapy , Aged, 80 and over , Anti-Bacterial Agents/therapeutic use , Blepharitis/diagnosis , Combined Modality Therapy/methods , Debridement/methods , Eye Infections, Bacterial/diagnosis , Fasciitis, Necrotizing/diagnosis , Female , Humans , Orbital Diseases/diagnosis , Treatment Outcome
9.
BMC Infect Dis ; 16: 314, 2016 06 30.
Article in English | MEDLINE | ID: mdl-27364885

ABSTRACT

BACKGROUND: Rapid diagnosis and appropriate antimicrobial therapy are of major importance to decrease morbidity and mortality in patients with blood stream infections (BSI). Blood culture, the current gold standard for detecting bacteria in blood, requires at least 24-48 hours and has limited sensitivity if obtained during antibiotic treatment of the patient. The aim of this prospective multicenter study was to clinically evaluate the application of a commercial universal 16S/18S rDNA PCR, SepsiTest™ (PCR-ST), directly on whole blood. METHODS: In total 236 samples from 166 patients with suspected sepsis were included in the study. PCR-ST results were compared to blood culture, the current gold standard for detecting BSI. Because blood cultures can give false-negative results, we performed an additional analysis to interpret the likelihood of bloodstream infection by using an evaluation based on clinical diagnosis, other diagnostic tests and laboratory parameters. RESULTS: Clinical interpretation of results defined the detected organism to be contaminants in 22 of 43 positive blood cultures (51.2 %) and 21 of 47 positive PCR-ST results (44.7 %). Excluding these contaminants resulted in an overall sensitivity and specificity of the PCR-ST of 66.7 and 94.4 % respectively. Of the 36 clinically relevant samples, 11 BSI were detected with both techniques, 15 BSI were detected with PCR-ST only and 10 with blood culture only. Therefore, in this study, SepsiTest™ detected an additional 71 % BSI compared to blood culture alone. CONCLUSIONS: More clinically relevant BSI were diagnosed by molecular detection, which might influence patient treatment. An improved SepsiTest™ assay suited for routine use can have additional value to blood culture in diagnosing bacteremia in septic patients.


Subject(s)
Bacteremia/diagnosis , Bacteria/genetics , DNA, Ribosomal/genetics , Polymerase Chain Reaction/methods , Adult , Aged , Aged, 80 and over , Bacteremia/microbiology , Blood Culture , Communicable Diseases , Female , Humans , Male , Middle Aged , Prospective Studies , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 18S/genetics , Sensitivity and Specificity , Sepsis/diagnosis , Sepsis/microbiology
10.
Haemophilia ; 17(5): 777-82, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21699628

ABSTRACT

Severe factor XI (sFXI) deficiency is a rare bleeding disorder (RBD). FXI replacement is most often required for surgical hemostasis. Plasma, the sole US treatment option, is often complicated by life-threatening allergic reactions. In such circumstances, the FDA offers a mechanism for institution-industry collaboration to facilitate limited use of replacement products licensed abroad. A 58 years old man with sFXI deficiency, required hip replacement. In the past, he received prophylactic plasma for thyroidectomy and experienced a severe allergic reaction. A single use institutional IND FDA application was initiated in collaboration with LFB (Les Ulis, France) to access Hemoleven®, a plasma-derived FXI concentrate. The application required an investigator-initiated IRB-approved protocol for treatment and safety/efficacy monitoring that included: preoperative thrombophilia, FXI inhibitor and pharmacokinetic (PK) evaluations; peri- postoperative administration of ≤ 4 doses of 10-15 U/kg Hemoleven® ; DIC monitoring; postoperative thromboprophylaxis; observation for product efficacy and potential complications. PK study demonstrated the expected 1.8% FXI recovery per U/kg with half-life of 62 hours. Mild D-Dimer elevation was noted 6-9 hours post-infusion. The initial dose (15 U/kg) was administered 15 hours before surgery; subsequently, 3 doses (10 U/kg) were infused every 72 hours. Hemostasis was excellent. No complications were observed. Collaboration allowed for successful patient access to Hemoleven® with excellent PK, safety, and efficacy. This case underscores the need for additional efforts to ensure safe and effective licensed replacement therapies for RBD patients.


Subject(s)
Anticoagulants/therapeutic use , Arthroplasty, Replacement, Hip , Factor XI Deficiency/drug therapy , Factor XI/therapeutic use , Arthroplasty, Replacement, Hip/methods , Blood Loss, Surgical/prevention & control , Hemostasis, Surgical/methods , Humans , Male , Middle Aged , Treatment Outcome
11.
Klin Padiatr ; 222(2): 73-8, 2010 Mar.
Article in English | MEDLINE | ID: mdl-19790029

ABSTRACT

BACKGROUND: Cat-scratch disease (CSD) is common in children, however the wide spectrum of the clinical presentation of CSD may lead to delayed diagnosis. An atypical presentation of CSD includes in its differential diagnosis diseases such as tuberculosis, other mycobacterioses, Epstein-Barr-Virus infection (EBV) or malignant disease. Since, in a small number of cases, these diseases may be present concurrently with an active CSD, it is important to consider CSD early in the differential diagnosis and order the appropriate tests. These tests include serology and, where possible, histology including molecular diagnostic methods on tissue specimens. PATIENTS AND METHOD: We performed a case series of five patients treated in our hospital with a clinical diagnosis of cat-scratch disease, confirmed by serology. An analysis of the history and clinical symptoms associated specifically with an atypical presentation of CSD was performed. RESULTS: The clinical presentation of CSD no longer encompasses the original typical description from 1950, but rather presents with a wide spectrum of signs and symptoms, including the absence of a documented cat scratch, fever, primary lesions or peripheral lymphadenopathy. Low density lesions in spleen, liver and lymph nodes are typical findings in ultrasound, MRI, or CT. Ignoring CSD as a possibility in investigating possible malignancy or tuberculosis could lead to unnecessary hospitalisation and delay in the proper treatment. CONCLUSION: CSD should also be considered in differential diagnosis of any patient with intraabdominal lymphadenopathy, abdominal pain and fever of unknown origin. A careful history is important, however, often patients with CSD have no history of contact with cats. Therefore in atypical cases of CSD the finding of other clinical symptoms and performance of specific diagnostic tests is important. Our experience suggests that early serological testing for Bartonella henselae should be performed and may avoid invasive diagnostic procedures.


Subject(s)
Bartonella henselae , Cat-Scratch Disease/diagnosis , Adolescent , Animals , Anti-Bacterial Agents/therapeutic use , Biopsy , Bites and Stings/complications , Blood Sedimentation , C-Reactive Protein/metabolism , Cat-Scratch Disease/drug therapy , Cats , Child , Child, Preschool , Diagnosis, Differential , Drug Therapy, Combination , Female , Humans , Laparoscopy , Liver/pathology , Lymph Nodes/pathology , Magnetic Resonance Imaging , Male , Spleen/pathology , Tomography, X-Ray Computed , Ultrasonography
12.
HSS J ; 4(1): 74-5, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18751867
13.
Bone Marrow Transplant ; 41(3): 253-9, 2008 Feb.
Article in English | MEDLINE | ID: mdl-17982498

ABSTRACT

Neurological complications are a relevant cause of morbidity and mortality after haematopoietic stem cell transplantation (SCT). We retrospectively analysed neurological complications of 165 paediatric patients who underwent SCT between 1996 and 2003. In all, 111 (67%) transplantations were allogeneic and 54 (33%) transplantations were autologous. Post-SCT neurological complications were seen in 24% of patients. They were seen in six children after autologous SCT and in 11 and 23 cases after allogeneic-related and -unrelated SCT. Neurological symptoms occurred between day +22 and +912 after transplantation and were classified into two groups. The first group (n=21) offered non-repetitive symptoms lasting less than 24 h without any cerebral imaging and cerebrospinal fluid(CSF) abnormalities. The second group (n=19) was characterized by progressive neurological symptoms, pathological MRI findings and/or abnormal results in CSF. Those with a progressive clinical course resulted from infections (n=10), drug toxicity (n=5), cerebrovascular events (n=2) and the central nervous system (CNS) relapse of the underlying disease (n=2). In particular, cerebral aspergillosis and toxoplasmosis after allogeneic unrelated SCT are a major challenge and are associated with a high mortality. In conclusion, our data suggest that patients presenting with progressive neurological symptoms after SCT require prompt diagnostic procedures and initiation in antimicrobial therapy in case of any findings suggestive of CNS infection.


Subject(s)
Hematopoietic Stem Cell Transplantation/adverse effects , Meningitis/etiology , Neoplasm Recurrence, Local , Neurotoxicity Syndromes/etiology , Seizures/etiology , Adolescent , Adult , Child , Child, Preschool , Cohort Studies , Female , Hospitals, Pediatric , Humans , Infant , Male , Meningitis/diagnosis , Meningitis/therapy , Neoplasm Recurrence, Local/complications , Neurotoxicity Syndromes/diagnosis , Neurotoxicity Syndromes/therapy , Retrospective Studies , Seizures/diagnosis , Seizures/therapy , Survival Analysis
14.
Klin Padiatr ; 219(6): 355-60, 2007.
Article in English | MEDLINE | ID: mdl-18050047

ABSTRACT

BACKGROUND: Children and adolescents after acute lymphoblastic leukemia are at risk for a prolonged period of immunodeficiency. Normally within 6 to 9 months after the end of maintenance treatment an adequate immune recovery is present. Factors such as immunity against specific antigens prior to disease (applied baseline vaccination), intensity of treatment and age can play a role in the appearance of antibodies in serum. Diphtheria (D) and Tetanus (T) antibodies are known to appear within 3 to 6 months after end of treatment as a sign of immune recovery and the reinstatement of immunological memory. A number of different questions are of interest: What differences are seen in the antibodies to D and T in children of different ages after treatment with a standardized protocol? What is the influence of post-treatment revaccination with Diphtheria/Tetanus (D/T) and treatment group on the production of D/T antibodies? PATIENTS AND METHODS: Out of 142 children and adolescents until the age of 16, treated according to the Co-ALL 05/92 protocol, 59 patients were eligible for evaluation: 31 Low-Risk (LR)- and 28 High-Risk (HR) patients. Antibodies against Diphtheria (D) and Tetanus (T) were measured 3-12 months after the end of treatment and after revaccination in case of low antibody levels against D and/or T. In patients without adequate response after repeated revaccination the cellular immunity was examined with a skin test. RESULTS: After the end of treatment, children in the low-risk (LR)-group showed more frequently adequate antibody titres against D and T than children of the high-risk (HR)-group. Antibodies against T were present in 50% of all patients. After revaccination antibodies against T were found in nearly all patients whereas for D this is only the case in some children. Patients without sufficient antibody levels mainly showed an adequate cellular immunity. CONCLUSION: In children and adolescents with ALL after therapy antibody levels of D and T are dependent on treatment intensity. Revaccination leads to an adequate immunological answer against T in most patients , which is not the case for the diphtheria vaccination. Prospective multicenter trials starting together with the ALL-treatment should be able to gain more information about the behavior of antibody levels and the risk of infection from vaccine-preventable disease in immunocompromised patients and thus lead to standardized vaccination guidelines such as immunization with conjugate vaccines already during maintenance treatment.


Subject(s)
Antibodies, Bacterial/blood , Diphtheria Toxoid/immunology , Immunologic Memory , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Precursor Cell Lymphoblastic Leukemia-Lymphoma/immunology , Tetanus Toxoid/immunology , Adolescent , Age Factors , Antineoplastic Combined Chemotherapy Protocols , Chi-Square Distribution , Child , Child, Preschool , Diphtheria/immunology , Diphtheria/prevention & control , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunity, Cellular , Male , Risk , Skin Tests , Tetanus/immunology , Tetanus/prevention & control , Time Factors , Vaccination
15.
Curr Drug Metab ; 8(3): 237-44, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17430112

ABSTRACT

Tryptophan metabolism occurs via the protohemoprotein enzymes tryptophan 2,3-dioxygenase (TDO) and indoleamine 2,3-dioxygenase (IDO), the latter action of which has a number of effects in the body including both antimicrobial defence and immune regulation. Whilst the antimicrobial action of IDO is largely due to depletion of the essential amino acid tryptophan, the immune regulatory function of IDO is still unclear and controversial. The list of pathogens that are "sensitive" to IDO-mediated tryptophan degradation covers intra-cellular parasites such as toxoplasma and possibly plasmodia, viruses (herpes viruses) to intra-cellular bacteria (chlamydia and rickettsia) and extra-cellular bacteria such as streptococci, enterococci and staphylococci. Immune regulation may be a consequence of tryptophan depletion, the accumulation of immune-active or toxic metabolites or due to other signalling events. This review covers the latest data and controversy pertaining to the antimicrobial and immune regulatory effects of tryptophan metabolism.


Subject(s)
Indoleamine-Pyrrole 2,3,-Dioxygenase/immunology , Animals , Anti-Infective Agents/immunology , Humans , Kynurenine/immunology , Kynurenine/metabolism , Kynurenine/toxicity , Tryptophan/immunology , Tryptophan/metabolism
16.
Histopathology ; 49(5): 515-22, 2006 Nov.
Article in English | MEDLINE | ID: mdl-17064298

ABSTRACT

AIMS: To characterize a single domain antibody (sdAb), AFAI, obtained by panning a naive phage display library of single domain antibodies against the non-small cell lung carcinoma cell line A549. AFAI recognizes a variant form of carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6 or CEA6). METHODS AND RESULTS: Various normal tissues and 139 neoplastic lesions from lung and other organs were immunostained with ES1 antibody, a pentameric and highly avid form of AFAI. ES1 was immunoreactive with 34 of 35 non-squamous large cell lung carcinomas with staining intensities ranging from focal to extensive and from moderate to strong. Importantly, ES1 stained poorly differentiated lung adenocarcinomas and many undifferentiated large cell lung carcinomas that typically show negative immunoreactivity with an antibody specific for thyroid transcription factor-1. Non-lung adenocarcinomas showed more focal and weaker immunoreactivity than for lung adenocarcinoma. Other carcinomas showed negative or weak immunoreactivity and normal tissues showed no immunoreactivity. CONCLUSIONS: Compared with other antibodies used in the clinical evaluation of lung adenocarcinomas, ES1 is more lung carcinoma sensitive, more sensitive in immunostaining of poorly differentiated adenocarcinomas that are usually associated with distant metastasis and less immunoreactive with normal tissues.


Subject(s)
Antibodies, Neoplasm/immunology , Antigens, CD/immunology , Antigens, Neoplasm/immunology , Carcinoma, Non-Small-Cell Lung/immunology , Cell Adhesion Molecules/immunology , Lung Neoplasms/immunology , Biomarkers, Tumor/immunology , Cell Line, Tumor , Female , GPI-Linked Proteins , Humans , Immunoenzyme Techniques , Male , Tissue Array Analysis
18.
J Clin Microbiol ; 43(1): 520-2, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15635034

ABSTRACT

Chlamydophila pneumoniae is mainly responsible for respiratory tract infections but has also been associated with endocarditis and myocarditis. We report a case of pneumonia in a child with hemorrhagic pericardial effusion with a positive result by a new C. pneumoniae TaqMan PCR, suggesting a pericardial inflammation directly induced by C. pneumoniae. C. pneumoniae should be suspected in patients with community-acquired pneumonia and concurrent pericarditis. Empirical treatment with azithromycin seems feasible.


Subject(s)
Chlamydophila Infections/complications , Chlamydophila pneumoniae/isolation & purification , Pericarditis/etiology , Pericarditis/microbiology , Pneumonia, Bacterial/complications , Acute Disease , Adolescent , Chlamydophila Infections/microbiology , Chlamydophila pneumoniae/classification , Chlamydophila pneumoniae/genetics , Female , Hemorrhage , Humans , Pericardial Effusion/microbiology , Pneumonia, Bacterial/microbiology
19.
Microbes Infect ; 6(9): 806-12, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15374002

ABSTRACT

Genital herpes simplex virus type 2 (HSV-2) is a significant clinical problem. Infection in pregnancy may result in disseminated infection of the newborn with encephalitis. We analyzed the antiviral effects induced by interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) in cervix carcinoma cells (HeLa) and astrocytoma cells (86HG39). We found that replication of HSV-2 in HeLa cells and in 86HG39 cells is inhibited after stimulation of the cells by IFN-gamma and TNF-alpha. The antiviral effect of IFN-gamma is enhanced in the presence of TNF-alpha, while stimulation by TNF-alpha alone did not induce antiviral activity. We found that IFN-gamma induces a strong activation of the tryptophan-degrading enzyme indoleamine 2,3-dioxygenase (IDO) and in addition, that the IFN-gamma-induced IDO activity was enhanced in the presence of TNF-alpha. Furthermore, we found that the induction of IDO activity is responsible for the inhibition of herpes simplex virus replication, since the presence of excess amounts of l-tryptophan abrogates the antiviral effect induced by IFN-gamma and the combination of IFN-gamma and TNF-alpha. We therefore conclude that the antiviral effect against HSV-2 mediated by type II interferon and TNF-alpha are dependent on IDO activation.


Subject(s)
Antiviral Agents/pharmacology , Herpesvirus 2, Human/drug effects , Interferon-gamma/pharmacology , Tryptophan Oxygenase/biosynthesis , Tumor Necrosis Factor-alpha/pharmacology , Cell Line, Tumor , Enzyme Activation , HeLa Cells , Herpesvirus 2, Human/pathogenicity , Herpesvirus 2, Human/physiology , Humans , Indoleamine-Pyrrole 2,3,-Dioxygenase , Recombinant Proteins , Tryptophan/metabolism , Virus Replication/drug effects
20.
Infection ; 32(2): 78-81, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15057571

ABSTRACT

BACKGROUND: Recent pediatric surveillance studies suggest the incidence of pneumococcal bacteremia, but not meningitis, is lower in Germany than in most developed countries. Suboptimal case assessment in routine clinical practice has been suspected of contributing to this apparent discrepancy. METHODS: We aimed to assess the blood culture sampling rate at a German pediatric university hospital and the disease burden associated with pneumococcal bacteremia in children under 5 years of age. The study design was retrospective, based on data-linkage and chart review. RESULTS: Blood cultures were frequently obtained in sepsis (96%; CI 78-99%) and meningitis (95%; CI 77-99%), but less commonly in pneumonia (49%; CI 43-54%) and fever without focus (48%; CI 38-59%). Pneumococci were the most common source of clinically significant bacteremia in previously healthy children. CONCLUSION: These blood culture sampling rates may be insufficient for the sensitive detection of pneumococcal bacteremia. Epidemiological surveillance based on poorly standardized diagnostic practices is prone to under-assessment.


Subject(s)
Bacteremia/epidemiology , Blood/microbiology , Pneumococcal Infections/epidemiology , Age Distribution , Bacteremia/diagnosis , Child , Child, Preschool , Female , Follow-Up Studies , Germany/epidemiology , Gram-Negative Bacteria/isolation & purification , Gram-Positive Bacteria/isolation & purification , Hospitals, Pediatric , Hospitals, University , Humans , Incidence , Infant , Male , Pneumococcal Infections/diagnosis , Retrospective Studies , Risk Factors , Sampling Studies , Sex Distribution , Survival Rate
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