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1.
Cancer Immunol Immunother ; 72(11): 3813-3824, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37742286

ABSTRACT

We investigated whether T cell-recruiting bispecific anti-CD3/GD2 antibody NG-CU might be an alternative to therapeutic anti-GD2 monoclonal antibody (mAb) ch14.18, mediating complement-dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) through natural killer (NK) cells for immunotherapy in high-risk/relapsed neuroblastoma after autologous/allogeneic stem cell transplantation (auto/alloSCT). Different antibody concentrations and effector-to-target ratios (E:T) were evaluated using xCELLigence RTCA system, peripheral blood mononuclear cells (PBMCs) (healthy donors and patients after alloSCT), and neuroblastoma cell lines (LS/LAN-1). Mean specific lysis of LS cells utilizing PBMCs from healthy donors and ch14.18 (1 µg/ml) was 40/66/75% after 12/24/48 h compared to 66/93/100% in the presence of NG-CU (100 ng/ml). NG-CU showed enhanced cytotoxicity compared to ch14.18, even at lower concentrations and E:T ratios, and completely eradicated LS cells after 72 h. To decipher the influence of effector cell subsets on lysis, different ratios of T and NK cells were tested. At a ratio of 1:1, ch14.18 was more effective than NG-CU. Using patient PBMCs taken at different time points posttransplant, significant lysis with both constructs was detectable depending on percentages and total numbers of T and NK cells; in the early posttransplant phase, NK cells were predominant and ch14.18 was superior, whereas later on, T cells represented the majority of immune cells and NG-CU was more effective. Our study highlights the importance of analyzing effector cell subsets in patients before initiating antibody-based therapy. Consequently, we propose an adjusted administration of both antibody constructs, considering the state of posttransplant immune recovery, to optimize anti-tumor activity.


Subject(s)
Antibodies, Bispecific , Antineoplastic Agents , Neuroblastoma , Humans , Leukocytes, Mononuclear/metabolism , Antibodies, Monoclonal/therapeutic use , Antineoplastic Agents/therapeutic use , Antibody-Dependent Cell Cytotoxicity , Neuroblastoma/drug therapy , Antibodies, Bispecific/pharmacology , Antibodies, Bispecific/therapeutic use , Gangliosides
2.
Leukemia ; 37(6): 1298-1310, 2023 06.
Article in English | MEDLINE | ID: mdl-37106163

ABSTRACT

Although the landscape for treating acute myeloid leukemia (AML) patients has changed substantially in recent years, the majority of patients will eventually relapse and succumb to their disease. Allogeneic stem cell transplantation provides the best anti-AML treatment strategy, but is only suitable in a minority of patients. In contrast to B-cell neoplasias, chimeric antigen receptor (CAR) T-cell therapy in AML has encountered challenges in target antigen heterogeneity, safety, and T-cell dysfunction. We established a Fab-based adapter CAR (AdCAR) T-cell platform with flexibility of targeting and control of AdCAR T-cell activation. Utilizing AML cell lines and a long-term culture assay for primary AML cells, we were able to demonstrate AML-specific cytotoxicity using anti-CD33, anti-CD123, and anti-CLL1 adapter molecules in vitro and in vivo. Notably, we show for the first time the feasibility of sequential application of adapter molecules of different specificity in primary AML co-cultures. Importantly, using the AML platform, we were able to demonstrate that chronic T-cell stimulation and exhaustion can be counteracted through introduction of treatment-free intervals. As T-cell exhaustion and target antigen heterogeneity are well-known causes of resistance, the AdCAR platform might offer effective strategies to ameliorate these limitations.


Subject(s)
Leukemia, Myeloid, Acute , T-Cell Exhaustion , Humans , Cell Line, Tumor , Leukemia, Myeloid, Acute/metabolism , Immunotherapy, Adoptive , T-Lymphocytes
3.
Front Neurol ; 13: 952699, 2022.
Article in English | MEDLINE | ID: mdl-36330424

ABSTRACT

Background and aim: Inflammatory myopathies are heterogeneous in terms of etiology, (immuno)pathology, and clinical findings. Endothelial cell injury, as it occurs in DM, is a common feature of numerous inflammatory and non-inflammatory vascular diseases. Vascular regeneration is mediated by both local and blood-derived mechanisms, such as the mobilization and activation of so-called proangiogenic cells (PACs) or early endothelial progenitor cells (eEPCs). The current study aimed to evaluate parameters of eEPC integrity in dermatomyositis (DM), compared to necrotizing myopathy (NM) and to non-myopathic controls. Methods: Blood samples from DM and NM patients were compared to non-myositis controls and analyzed for the following parameters: circulating CD133+/VEGFR-2+ cells, number of colony-forming unit endothelial cells (CFU-ECs), concentrations of angiopoietin 1, vascular endothelial growth factor (VEGF), and CXCL-16. Muscle biopsies from DM and NM subjects underwent immunofluorescence analysis for CXCR6, nestin, and CD31 (PECAM-1). Finally, myotubes, derived from healthy donors, were stimulated with serum samples from DM and NM patients, subsequently followed by RT-PCR for the following candidates: IL-1ß, IL-6, nestin, and CD31. Results: Seventeen (17) DM patients, 7 NM patients, and 40 non-myositis controls were included. CD133+/VEGFR-2+ cells did not differ between the groups. Both DM and NM patients showed lower CFU-ECs than controls. In DM, intramuscular CD31 abundances were significantly reduced, which indicated vascular rarefaction. Muscular CXCR6 was elevated in both diseases. Circulating CXCL-16 was higher in DM and NM in contrast, compared to controls. Serum from patients with DM but not NM induced a profound upregulation of mRNS expression of CD31 and IL-6 in cultured myotubes. Conclusion: Our study demonstrates the loss of intramuscular microvessels in DM, accompanied by endothelial activation in DM and NM. Vascular regeneration was impaired in DM and NM. The findings suggest a role for inflammation-associated vascular damage in the pathogenesis of DM.

4.
J Pediatr Urol ; 18(2): 132-140, 2022 04.
Article in English | MEDLINE | ID: mdl-35148953

ABSTRACT

INTRODUCTION: Bladder stones (BS) are still endemic in children in developing nations and account for a high volume of paediatric urology workload in these areas. The aim of this systematic review is to comparatively assess the benefits and risks of minimally invasive and open surgical interventions for the treatment of bladder stones in children. METHODS: This systematic review was conducted in accordance with Cochrane Guidance. Database searches (January 1970- March 2021) were screened, abstracted, and assessed for risk of bias for comparative randomised controlled trials (RCTs) and non-randomised studies (NRSs) with >10 patients per group. Open cystolithotomy (CL), transurethral cystolithotripsy (TUCL), percutaneous cystolithotripsy (PCCL), extracorporeal shock wave lithotripsy (ESWL) and laparoscopic cystolithotomy (LapCL) were evaluated. RESULTS: In total, 3040 abstracts were screened, and 8 studies were included. There were 7 retrospective non-randomised studies (NRS's) and 1 quasi-RCT with 1034 eligible patients (CL: n=637, TUCL: n=196, PCCL: n=138, ESWL: n=63, LapCL n=0). Stone free rate (SFR) was given in 7 studies and measured 100%, 86.6%-100%, and 100% for CL, TUCL and PCCL respectively. CL was associated with a longer duration of inpatient stay than PCCL and TUCL (p<0.05). One NRS showed that SFR was significantly lower after 1 session with outpatient ESWL (47.6%) compared to TUCL (93.5%) and CL (100%) (p<0.01 and p<0.01 respectively). One RCT compared TUCL with laser versus TUCL with pneumatic lithotripsy and found that procedure duration was shorter with laser for stones <1.5cm (n=25, p=0.04). CONCLUSION: In conclusion, CL, TUCL and PCCL have comparable SFRs but ESWL is less effective for treating stones in paediatric patients. CL has the longest duration of inpatient stay. Information gathered from this systematic review will enable paediatric urologists to comparatively assess the risks and benefits of all urological modalities when considering surgical intervention for bladder stones.


Subject(s)
Lithotripsy , Urinary Bladder Calculi , Urology , Child , Developing Countries , Humans , Lithotripsy/methods , Treatment Outcome , Urinary Bladder/surgery , Urinary Bladder Calculi/surgery
5.
Urologe A ; 59(12): 1498-1503, 2020 Dec.
Article in German | MEDLINE | ID: mdl-33237370

ABSTRACT

Increase of medical knowledge, technical innovation together with a demographic change, and increase of stone incidence in daily practice challenges guideline preparation and clinical studies. Increasing interdisciplinary collaboration in stone treatment can also be demonstrated in the number of affiliated professional and working groups in the current guideline update. The following case illustrates treatment options in a symptomatic patient harbouring bilateral stones and metabolic risk factors. Decision guidance for treatment and recurrence prevention measures are presented on the basis of expert opinion and available published evidence.


Subject(s)
Kidney Calculi , Lithotripsy , Urolithiasis , Humans , Recurrence , Treatment Outcome , Urolithiasis/therapy
6.
World J Urol ; 38(3): 761-768, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31093704

ABSTRACT

PURPOSE: Fluoroscopy is valuable in modern endourology. We present the results of a survey where compliance to radiation safety measures was tested according to surgical exposure, and level of understanding of the radiation rules and risks associated with it. METHODS: A 52-item, anonymous questionnaire, structured by 6 ESUT/EULIS experts was distributed at 3 different endourological meetings during 2017-2018. Main aim was to evaluate level of knowledge on radiation physics and the protective measures taken against radiation exposure by participants. Fisher's exact test, Kruskal-Wallis test and ROC curve were used for statistical analysis. RESULTS: 211 responses were evaluated. Number of correct answers (median 7.00) differed significantly according to age (p = 0.001), working position (p = 0.005), working field (p < 0.001), number of semirigid (p < 0.001)/flexible URS (p < 0.001) and PNL (p < 0.001) performed per year. Physicians aged 50-60 years, consultants, academics and those who performed more procedures achieved higher scores. In our study 51.7% of responders used shields in the operating room, 89.6% wore lead aprons, 84.4% thyroid shields, while glasses and gloves were used by 14.7% and 8.1%, respectively. Age, working field and number of correct answers did not affect significantly the use of protection in contrast with endourology fellowship training, working position and lessons on radiation. Interestingly, residents, untrained endourologists and those who were provided with lessons on radiation were more compliant. CONCLUSIONS: Our study revealed that majority of modern urologists advocate radiation protection during endourology practice. Senior consultants and academic urologists performing a high volume of procedures seem to understand physics and rules of radiation use.


Subject(s)
Endoscopy , Occupational Exposure/prevention & control , Personal Protective Equipment , Professional Competence , Radiation Exposure/prevention & control , Urologists , Academic Medical Centers , Adult , Age Factors , Female , Fluoroscopy/adverse effects , Guideline Adherence , Humans , Male , Middle Aged , Private Sector , Public Sector , Surveys and Questionnaires
7.
Urologe A ; 58(11): 1304-1312, 2019 Nov.
Article in German | MEDLINE | ID: mdl-31506761

ABSTRACT

The increase of medical knowledge and technical innovations together with the demographic change represent a challenge for the new conception of guidelines and clinical studies. The present S2k guidelines, which are exclusively concerned with kidney and ureteral stones, should support the treatment of urolithiasis in hospitals and private practices and provide information on urolithiasis for patients. Increasing interdisciplinary collaboration in stone treatment is also demonstrated in the number of professional and working groups participating in the update of the new guidelines. The present S2k guidelines emerged from a consensus process and demonstrate the current recommendations in step with actual practice. They provide decision-making guidance for diagnostics, treatment and metaphylactic measures based on expert opinions and available published fundamental evidence from the literature.


Subject(s)
Lithotripsy/standards , Practice Guidelines as Topic , Ureteroscopy/standards , Urolithiasis/surgery , Urologic Surgical Procedures/standards , Urology/standards , Extracorporeal Shockwave Therapy , Humans , Kidney Calculi , Nephrolithotomy, Percutaneous , Treatment Outcome , Ureteral Calculi , Urolithiasis/diagnosis , Urolithiasis/prevention & control , Urologic Surgical Procedures/instrumentation
8.
Eur J Nucl Med Mol Imaging ; 45(2): 235-242, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29075832

ABSTRACT

OBJECTIVE: To evaluate the diagnostic performance of [68Ga]Ga-PSMAHBED-CC conjugate 11 positron emission tomography (PSMA-PET) in the early detection of metastases in patients with biochemical recurrence (BCR) after radical prostatectomy (RP) for clinically non-metastatic prostate cancer, to compare it to CT/MRI alone and to assess its impact on further therapeutic decisions. MATERIAL AND METHODS: We retrospectively assessed 117 consecutive hormone-naïve BCR patients who had 68Ga-PSMA 11 PET/CT (n = 46) or PET/MRI (n = 71) between May 2014 and January 2017. BCR was defined as two PSA rises above 0.2 ng/ml. Two dedicated uro-oncological imaging experts (radiology/nuclear medicine) reviewed separately all images. All results were presented in a blinded sequential fashion to a multidisciplinary tumorboard in order to assess the influence of PSMA-PET imaging on decision-making. RESULTS: The median time from RP to BCR was 36 months (IQR 16-72). Overall, 69 (59%) patients received postoperative radiotherapy. Median PSA level at the time of imaging was 1.04 ng/ml (IQR 0.58-1.87). PSMA-positive lesions were detected in 100 (85.5%) patients. Detection rates were 65% for a PSA value of 0.2 to <0.5 ng/ml, 85.7% for 0.5 to <1, 85.7% for 1 to <2 and 100% for ≥2. PSMA-positive lesions could be confirmed by either histology (16%), PSA decrease in metastasis-directed radiotherapy (45%) or additional information in diffusion-weighted imaging when PET/MRI was performed (18%) in 79% of patients. PSMA-PET detected lesions in 67 patients (57.3%) who had no suspicious correlates according to the RECIST 1.1 criteria on MRI or CT. PSMA-PET changed therapeutic decisions in 74.6% of these 67 patients (p < 0.001), with 86% of them being considered for metastases-directed therapies. CONCLUSIONS: We confirm the high performance of PSMA-PET imaging for the detection of disease recurrence sites in patients with BCR after RP, even at relatively low PSA levels. Moreover, it adds significant information to standard CT/MRI, changing treatment strategies in a significant number of patients.


Subject(s)
Decision Making , Edetic Acid/analogs & derivatives , Oligopeptides/metabolism , Positron-Emission Tomography , Prostatectomy , Prostatic Neoplasms/diagnostic imaging , Prostatic Neoplasms/metabolism , Aged , Edetic Acid/metabolism , Gallium Isotopes , Gallium Radioisotopes , Humans , Ligands , Male , Prostatic Neoplasms/pathology , Prostatic Neoplasms/therapy , Recurrence , Retrospective Studies
9.
Z Rheumatol ; 76(5): 382-390, 2017 Jun.
Article in German | MEDLINE | ID: mdl-28314915

ABSTRACT

Skin manifestations of sarcoidosis occur in up to 30% of cases. This review summarizes and illustrates in detail the differences between specific and unspecific skin manifestations of sarcoidosis. Important differential diagnoses, such as tuberculosis, cutaneous lymphoma and syphilis have to be excluded. The indications for systemic treatment are primarily determined by the extent of organ involvement and also by the cosmetic impairment.


Subject(s)
Sarcoidosis/diagnosis , Sarcoidosis/therapy , Skin Diseases/diagnosis , Skin Diseases/therapy , Diagnosis, Differential , Evidence-Based Medicine , Humans , Treatment Outcome
10.
Prostate Cancer Prostatic Dis ; 20(2): 197-202, 2017 06.
Article in English | MEDLINE | ID: mdl-28071673

ABSTRACT

BACKGROUND: We aimed to assess the prognostic relevance of the new Grade Groups in Prostate Cancer (PCa) within a large cohort of European men treated with radical prostatectomy (RP). METHODS: Data from 27 122 patients treated with RP at seven European centers were analyzed. We investigated the prognostic performance of the new Grade Groups (based on Gleason score 3+3, 3+4, 4+3, 8 and 9-10) on biopsy and RP specimen, adjusted for established clinical and pathological characteristics. Multivariable Cox proportional hazards regression models assessed the association of new Grade Groups with biochemical recurrence (BCR). Prognostic accuracies of the models were assessed using Harrell's C-index. RESULTS: Median follow-up was 29 months (interquartile range, 13-54). The 4-year estimated BCR-free survival (bRFS) for biopsy Grade Groups 1-5 were 91.3, 81.6, 69.8, 60.3 and 44.4%, respectively. The 4-year estimated bRFS for RP Grade Groups 1-5 were 96.1%, 86.7%, 67.0%, 63.1% and 41.0%, respectively. Compared with Grade Group 1, all other Grade Groups based both on biopsy and RP specimen were independently associated with a lower bRFS (all P<0.01). Adjusted pairwise comparisons revealed statistically differences between all Grade Groups, except for group 3 and 4 on RP specimen (P=0.10). The discriminations of the multivariable base prognostic models based on the current three-tier and the new five-tier systems were not clinically different (0.3 and 0.9% increase in discrimination for clinical and pathological model). CONCLUSIONS: We validated the independent prognostic value of the new Grade Groups on biopsy and RP specimen from European PCa men. However, it does not improve the accuracies of prognostic models by a clinically significant margin. Nevertheless, this new classification may help physicians and patients estimate disease aggressiveness with a user-friendly, clinically relevant and reproducible method.


Subject(s)
Neoplasm Grading/methods , Prognosis , Prostatic Neoplasms/epidemiology , Prostatic Neoplasms/pathology , Aged , Biopsy , Europe , Humans , Kaplan-Meier Estimate , Male , Middle Aged , Neoplasm Recurrence, Local/epidemiology , Neoplasm Recurrence, Local/pathology , Predictive Value of Tests , Proportional Hazards Models , Prostate/pathology , Prostatic Neoplasms/classification
11.
J Reprod Immunol ; 119: 9-14, 2017 02.
Article in English | MEDLINE | ID: mdl-27865124

ABSTRACT

Peripheral and uterine NK cells (pNK, uNK) can be distinguished according to their receptor expression. Recent studies indicate an association of elevated pNK and uNK with recurrent miscarriage (RM). This study aimed to analyze pNK and uNK in patients with RM and healthy controls. Out of n=590 RM patients screened according to a standard diagnostic protocol, n=268 couples with ≥3 consecutive RM were identified. Subgroups consisted of n=151 primary RM (pRM), n=85 secondary RM (sRM), n=32 tertiary RM (tRM) and n=42 healthy controls. Finally, n=147 idiopathic RM (iRM) and n=121 non-iRM patients were identified. Peripheral blood levels of CD45+CD3-CD56+CD16+ NK cells were determined in non-pregnant patients and controls in the mid-luteal phase by FACS. In n=129 RM patients a uterine biopsy was taken to evaluate CD56+ NK cells by immunohistochemistry. PRM showed higher absolute pNK than sRM (median/µl (Q1;Q3): 234 (147;306) vs 176 (128;245), p=0.02). Further a trend towards higher pNK percentages in pRM was detected. UNK numbers did not differ between RM subgroups and did not correlate with pNK. However, the rate of highly elevated uNK was increased in iRM compared to non-iRM patients (p=0.04). Further, higher numbers of CD45+CD3-DR+ (p<0.01) and CD45+CD3+CD8+DR+ (p=0.04) peripheral lymphocytes were associated with higher uNK numbers. In conclusion, elevated pNK were present in pRM patients. Although pNK and uNK numbers did not correlate, the association between high CD45+CD3-DR+ and CD45+CD3+CD8+DR+ peripheral lymphocytes and uNK might indicate that activated NK, B and T cells provide cytokines for the differentiation of uNK.


Subject(s)
Abortion, Habitual/immunology , Blood Cells/immunology , Killer Cells, Natural/immunology , Uterus/pathology , Adult , Antigens, CD/metabolism , B-Lymphocytes/immunology , Cells, Cultured , Female , Humans , Lymphocyte Activation , Male , Paracrine Communication , Pregnancy , T-Lymphocytes/immunology
13.
BMC Musculoskelet Disord ; 17: 339, 2016 08 12.
Article in English | MEDLINE | ID: mdl-27519706

ABSTRACT

BACKGROUND: Patients with systemic sclerosis (SSc) are endagered by tissue fibrosis and by microvasculopathy, with the latter caused by endothelial cell expansion/proliferation. SSc-associated fibrosis potentially results from mesenchymal transdifferentiation of endothelial cells. Early Endothelial Progenitor Cells (eEPCs) act proangiogenic under diverse conditions. Aim of the study was to analyze eEPC regeneration and mesenchymal transdifferentiation in patients with limited and diffuse SSs (lSSc and dSSc). METHODS: Patients with both, lSSc and dSSc were included into the study. The following parameters were evaluated: eEPC numbers and regeneration, concentrations of vasomodulatory mediators, mesenchymal properties of blood-derived eEPC. Serum samples of healthy subjects and SS patients were used for stimulation of cultured human eEPC, subsequently followed by analysis of mesenchymal cell characteristics and mobility. RESULTS: Twenty-nine patients were included into the study. Regenerative activity of blood-derived eEPCs did not differ between Controls and patients. Circulating eEPC were significantly lower in all patients with SSc, and in limited and diffuse SSc (lSSc/dSSc). Serum concentrations of promesenchymal TGF-b was elevated in all patients with SSc. Cultured mononuclear cells from SS patients displayed higher abundances of CD31 and of CD31 and aSMA combined. Finally, serum from SSc patients inhibited migration of cultured eEPCs and the cells showed lower sensitivity towards the endothelin antagonist Bosentan. CONCLUSIONS: The eEPC system, which represents an essential element of the endogenous vascular repair machinery is affected in SSc. The increased appearance of mesenchymal properties in eEPC may indicate that alterations of the cells potentially contribute to the accumulation of connective tissue and to vascular malfunction.


Subject(s)
Cell Transdifferentiation , Endothelial Progenitor Cells/physiology , Scleroderma, Diffuse/etiology , Scleroderma, Limited/etiology , Biomarkers/blood , Case-Control Studies , Cell Movement , Female , Humans , Male , Middle Aged , Prospective Studies , Regeneration , Scleroderma, Diffuse/blood , Scleroderma, Limited/blood
14.
Prostate Cancer Prostatic Dis ; 19(2): 163-7, 2016 06.
Article in English | MEDLINE | ID: mdl-26810014

ABSTRACT

BACKGROUND: To assess the prognostic value of preoperative C-reactive protein (CRP) serum levels for prognostication of biochemical recurrence (BCR) after radical prostatectomy (RP) in a large multi-institutional cohort. METHODS: Data from 7205 patients treated with RP at five institutions for clinically localized prostate cancer (PCa) were retrospectively analyzed. Preoperative serum levels of CRP within 24 h before surgery were evaluated. A CRP level ⩾0.5 mg dl(-1) was considered elevated. Associations of elevated CRP with BCR were evaluated using univariable and multivariable Cox proportional hazards regression models. Harrel's C-index was used to assess prognostic accuracy (PA). RESULTS: Patients with higher Gleason score on biopsy and RP, extracapsular extension, seminal vesicle invasion, lymph node metastasis, and positive surgical margins status had a significantly elevated preoperative CRP compared to those without these features. Patients with elevated CRP had a lower 5-year BCR survival proportion as compared to those with normal CRP (55% vs 76%, respectively, P<0.0001). In pre- and postoperative multivariable models that adjusted for standard clinical and pathologic features, elevated CRP was independently associated with BCR (P<0.001). However, the addition of preoperative CRP did not improve the accuracy of the standard pre- and postoperative models for prediction of BCR (70.9% vs 71% and 78.9% vs 78.7%, respectively). CONCLUSIONS: Preoperative CRP is elevated in patients with pathological features of aggressive PCa and BCR after RP. While CRP has independent prognostic value, it does not add prognostically or clinically significant information to standard predictors of outcomes.


Subject(s)
C-Reactive Protein , Prostatic Neoplasms/blood , Prostatic Neoplasms/diagnosis , Aged , Follow-Up Studies , Humans , Kaplan-Meier Estimate , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Grading , Preoperative Period , Prognosis , Proportional Hazards Models , Prostatectomy , Prostatic Neoplasms/mortality , Prostatic Neoplasms/surgery , Recurrence
15.
Mol Biosyst ; 12(6): 1750-5, 2016 05 24.
Article in English | MEDLINE | ID: mdl-26776361

ABSTRACT

The covalent attachment of polyethylene glycol (PEG) to therapeutic proteins can improve their physicochemical properties. In this work we utilized the non-natural amino acid p-azidophenylalanine (pAzF) in combination with the chemoselective Staudinger-phosphite reaction to install branched PEG chains to recombinant unglycosylated erythropoietin (EPO) at each single naturally occurring glycosylation site. PEGylation with two short 750 or 2000 Da PEG units at positions 24, 38, or 83 significantly decreased unspecific aggregation and proteolytic degradation while biological activity in vitro was preserved or even increased in comparison to full-glycosylated EPO. This site-specific bioconjugation approach permits to analyse the impact of PEGylation at single positions. These results represent an important step towards the engineering of site-specifically modified EPO variants from bacterial expression with increased therapeutic efficacy.


Subject(s)
Erythropoietin/chemistry , Erythropoietin/metabolism , Polyethylene Glycols/chemistry , Recombinant Proteins , Amino Acid Sequence , Amino Acids/chemistry , Amino Acids/metabolism , Animals , Cell Line , Cell Proliferation/drug effects , Erythropoietin/genetics , Erythropoietin/pharmacology , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Glycosylation , Humans , Molecular Structure , Protein Conformation , Protein Stability
16.
Clin Exp Allergy ; 46(1): 125-32, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26767493

ABSTRACT

BACKGROUND: Immunoglobulin E-mediated allergy to drugs and substances used during general anaesthesia as well as non-allergic drug hypersensitivity reactions may account for anaesthesia-induced anaphylaxis. As IgE-mediated anaphylaxis is a potentially life-threatening reaction, identification of the culprit allergen is essential to avoid anaphylaxis recurrence during subsequent general anaesthesia. OBJECTIVE: To study whether preventive recommendations derived from allergy testing after intraoperative anaphylaxis were followed in subsequent general anaesthesia. METHODS: Results of standardized allergy testing after anaesthesia-induced anaphylaxis and outcome of subsequent general anaesthesia were analysed retrospectively. RESULTS: Fifty-three of 107 patients were diagnosed with IgE-mediated allergy to a drug or substance used during general anaesthesia, and 54 patients were test negative. Twenty-eight of 29 allergy patients tolerated subsequent general anaesthesia uneventfully. One patient with cefazolin allergy suffered from anaphylaxis recurrence due to accidental reapplication of cefazolin. Twenty-two of 24 test-negative patients tolerated subsequent general anaesthesia, whereas two patients again developed anaphylaxis despite pre-medication regimens. CONCLUSION AND CLINICAL RELEVANCE: Our results confirm the practical impact of allergy testing in general anaesthesia-induced anaphylaxis. By identification of the allergen, it is possible to avoid allergic anaphylaxis during subsequent anaesthesia. In most cases, recommended pre-medication seems to prevent the recurrence of non-allergic drug hypersensitivity reactions.


Subject(s)
Anaphylaxis/etiology , Anesthesia, General/adverse effects , Adolescent , Adult , Aged , Aged, 80 and over , Allergens/immunology , Anaphylaxis/diagnosis , Anaphylaxis/prevention & control , Antibody Specificity/immunology , Basophils/immunology , Basophils/metabolism , Child , Drug Hypersensitivity/diagnosis , Drug Hypersensitivity/immunology , Female , Humans , Immunization , Immunoglobulin E/blood , Immunoglobulin E/immunology , Male , Middle Aged , Retrospective Studies , Skin Tests , Tryptases/blood , Workflow , Young Adult
19.
Klin Padiatr ; 227(4): 219-24, 2015 Jul.
Article in German | MEDLINE | ID: mdl-25811742

ABSTRACT

BACKGROUND: Metabolic acidosis is a common problem of patients on neonatal intensive care units. Only little data exists in literature and there are no clinical guidelines. The aim of this national survey was to assess criteria for correction of metabolic acidosis in neonatal patients and if there were effects to be observed. METHODS: We designed an online survey and sent it to 304 German children's hospitals. 101 questionnaires were included in our study. RESULTS: The question "How often do you buffer on your ward a week?" was answered 63 times with "zero". In perinatal asphyxia newborns with gestation age over 36+0 weeks 4% of the neonatologists would frequently perform a correction of acidosis, 74.3% would do it rarely and 21.8% never. In syndrome of persistent fetal circulation 28.4% would correct acidosis frequently, 42.0% would correct it rarely and 29.5% would never correct it. In case of sepsis 8.7% would correct acidosis frequently, 70.7% would do it rarely and 20.7% would never correct it. 75.2% of the participants distinguish in buffering a premature or a mature infant. 44.4% of neonatologists saw an improvement of the clinical status of the patient after buffering. 38.3% saw different effects, 16.0% saw no changes and 1.2% saw a worsening of the clinical status. 49.4% of those questioned saw side effects after using sodium bicarbonate as a buffer. CONCLUSION: Correction of acidosis with a buffer is rarely performed on German neonatology wards. The indication of buffering depends on the clinical picture and its underlying problem. Benefits from buffering were seen, as well as side effects.


Subject(s)
Acidosis/therapy , Infant, Premature, Diseases/therapy , Intensive Care Units, Neonatal , Practice Patterns, Physicians'/statistics & numerical data , Acidosis/diagnosis , Asphyxia Neonatorum/diagnosis , Asphyxia Neonatorum/therapy , Buffers , Female , Germany , Gestational Age , Health Services Research , Humans , Infant, Newborn , Infant, Premature, Diseases/diagnosis , Internet , Male , Persistent Fetal Circulation Syndrome/diagnosis , Persistent Fetal Circulation Syndrome/therapy , Sepsis/diagnosis , Sepsis/therapy , Sodium Bicarbonate/administration & dosage , Sodium Bicarbonate/adverse effects , Surveys and Questionnaires , Treatment Outcome
20.
World J Urol ; 33(9): 1297-302, 2015 Sep.
Article in English | MEDLINE | ID: mdl-25385490

ABSTRACT

PURPOSE: To investigate the impact of gender differences on treatment success, intraoperative and postoperative complications in patients undergoing ureteroscopy (URS). MATERIALS AND METHODS: A prospectively maintained database of 927 consecutively performed ureteroscopies on solitary ureteral stones in four different centers was retrospectively analyzed. Stones were detected with preoperative computed tomography scans or intravenous urography imaging. Patients received intravenous antibiotics as perioperative prophylaxis. Patients with symptomatic urinary tract infections (UTI) prior to surgery were excluded. Follow-up was up to 2 weeks after URS or stent removal. RESULTS: Two hundred and eighty-six women and 641 men were included in this study. Mean stone size was 9 mm (range 2-35 mm). A double-J stent was placed in 240 (83 %) women and 527 (82 %) men at the end of surgery (p = 0.075). There was no significant gender difference in terms of stent dislocation (p = 0.239). Two hundred and fifty-one women (87 %) and 564 men (87 %) were stone-free after the first procedure (p = 0.917). Intraoperative complications were observed in 14 (4.8 %) women and 37 (5.9 %) men (p = 0.313). Severe UTI presenting with fever (>38 °C) and requiring prolonged hospitalization with parenteral antibiotics were observed in 11 (3 %) women and 8 (1 %) men postoperatively. This difference was statistically significant (p = 0.025). CONCLUSION: No significant differences between female and male patients harboring ureteral stones with respect to intraoperative complications were detected. Although stone characteristics were comparable between groups, a small number of women had significantly more severe UTI's postoperatively. Our current therapy regimen for URS seems to be efficient and safe both for females and males.


Subject(s)
Postoperative Complications/epidemiology , Ureteral Calculi/surgery , Ureteroscopy , Adolescent , Adult , Aged , Aged, 80 and over , Austria/epidemiology , Croatia/epidemiology , Female , Follow-Up Studies , Humans , Incidence , Male , Middle Aged , Retrospective Studies , Sex Distribution , Sex Factors , Tomography, X-Ray Computed , Treatment Outcome , Turkey/epidemiology , Ureteral Calculi/diagnosis , Young Adult
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