Prospective evaluation of ultrasound features of magnesium-based bioabsorbable screw resorption in pediatric fractures.

OBJECTIVE: Bioabsorbable magnesium-based alloy screws release gas upon resorption. The resulting findings in the adjacent soft tissues and joints may mimic infection. The aim of the study was to evaluate the ultrasound (US) findings in soft tissues and joints during screw resorption. METHODS: Prospectively acquired US studies from pediatric patients treated with magnesium screws were evaluated for screw head visibility, posterior acoustic shadowing, twinkling artifact, foreign body granuloma, gas (soft tissue, intra-articular), alterations of the skin and subcutaneous fat, perifascial fluid, localized fluid collections, hypervascularization, and joint effusion. RESULTS: Sixty-six US studies of 28 pediatric patients (nfemale = 9, nmale = 19) were included. The mean age of the patients at the time of surgery was 10.84 years; the mean time between surgery and ultrasound was 128.3 days (range = 6-468 days). The screw head and posterior acoustic shadowing were visible in 100% of the studies, twinkling artifact in 6.1%, foreign body granuloma in 92.4%, gas locules in soft tissue in 100% and intra-articular in 18.2%, hyperechogenicity of the subcutaneous fat in 90.9%, cobblestoning of the subcutaneous fat in 24.2%, loss of normal differentiation between the epidermis/dermis and the subcutaneous fat in 57.6%, localized fluid collection in 9.9%, perifascial fluid in 12.1%, hypervascularization in 27.3%, and joint effusion in 18.2%. CONCLUSION: US findings in pediatric patients treated with magnesium screws strongly resemble infection, but are normal findings in the setting of screw resorption. CLINICAL RELEVANCE STATEMENT: Bioabsorbable magnesium-based alloy screws release gas during resorption. The resulting US findings in the adjacent soft tissues and joints in pediatric patients may mimic infection, but are normal findings. KEY POINTS: • Bioabsorbable magnesium-based alloy screws release gas upon resorption. • The resulting ultrasound findings in children's soft tissues and joints closely resemble those of soft tissue infection or osteosynthesis-associated infection. • Be familiar with these ultrasound findings in order to avoid inadvertently misdiagnosing a soft tissue infection or osteosynthesis-associated infection.

Diagnostic performance of long head of biceps tendon tears on MRI: systematic review and meta-analysis.

OBJECTIVES: The purpose of this meta-analysis was to determine the diagnostic performance of conventional MRI and MR arthrography for tendinosis, and partial and complete tears of the long head of the biceps tendon (LHBT) using arthroscopy as the reference standard. MATERIALS AND METHODS: A systematic review was performed using predefined data fields in PubMed, and all articles published from January 2000 up to April 2022 were retrospectively pooled and reviewed. Six MRI studies on complete tear (n = 555) and ten studies on partial tear/tendinosis (n = 2487) were included in the analysis. Two of the included studies in each group investigated the use of MR arthrography. The data sets were analyzed using a univariate approach with the DerSimonian and Laird random effects model and the proportional hazards model. RESULTS: MRI shows high specificities in diagnosing complete tears of the LHBT ranging from 93.0 to 99.0%. Diagnostic sensitivity was more heterogeneous ranging from 55.9 to 90.0%. The overall negative likelihood ratio was 0.29 (95% CI: 0.17-0.50) and the overall positive likelihood ratio was 37.3 (95% CI: 11.9-117.4). The mean sensitivity in diagnosing partial tear/tendinosis of the LHBT was 67.8% (95% CI: 54.3-78.9%) and the specificity was 75.9% (95% CI: 63.6-85.0%), resulting in a balanced accuracy of 71.9%. The overall negative likelihood ratio was 0.44 (95% CI: 0.32-0.59) and the overall positive likelihood ratio was 2.64 (95% CI: 1.91-3.65). CONCLUSION: MRI is highly specific for the diagnosis of complete tears of the LHBT, whereas diagnostic sensitivity was more heterogeneous. The diagnosis of partial tears and/or tendinosis of the LHBT remains challenging on MRI, which may warrant complementary clinical examination or other imaging modalities to increase diagnostic confidence in equivocal cases. CLINICAL RELEVANCE STATEMENT: Conventional MRI and MR arthrography have high diagnostic performance for complete tendon tear when compared to arthroscopy. The diagnosis of tendinosis/partial tears remains challenging and may require comparison with clinical tests and other imaging modalities. KEY POINTS: •There is no clear consensus regarding the primary imaging modality for the evaluation of LHBT disorders. •Conventional MRI and MR arthrography are highly specific in diagnosing complete tears of the LHBT. •Diagnosis of partial tears/tendinosis of the LHBT on conventional MRI and MR arthrography remains a diagnostic challenge.

Covalent S-Adenosylhomocysteine-Based DNA Methyltransferase 2 Inhibitors with a New Type of Aryl Warhead.

The DNA methyltransferase 2 (DNMT2) is an RNA modifying enzyme associated with pathophysiological processes, such as mental and metabolic disorders or cancer. Although the development of methyltransferase inhibitors remains challenging, DNMT2 is not only a promising target for drug discovery, but also for the development of activity-based probes. Here, we present covalent SAH-based DNMT2 inhibitors decorated with a new type of aryl warhead. Based on a noncovalent DNMT2 inhibitor with N-benzyl substituent, the Topliss scheme was followed for optimization. The results showed that electron-deficient benzyl moieties highly increased affinity. By decorating the structures with strong electron-withdrawing moieties and leaving groups, we adjusted the electrophilicity to create covalent DNMT2 inhibitors. A 4-bromo-3-nitrophenylsulfonamide-decorated SAH derivative (80) turned out to be the most potent (IC50 = 1.2 ± 0.1 µM) and selective inhibitor. Protein mass spectrometry confirmed the covalent reaction with the catalytically active cysteine-79.

Discovery of Inhibitors of DNA Methyltransferase 2, an Epitranscriptomic Modulator and Potential Target for Cancer Treatment.

Selective manipulation of the epitranscriptome could be beneficial for the treatment of cancer and also broaden the understanding of epigenetic inheritance. Inhibitors of the tRNA methyltransferase DNMT2, the enzyme catalyzing the S-adenosylmethionine-dependent methylation of cytidine 38 to 5-methylcytidine, were designed, synthesized, and analyzed for their enzyme-binding and -inhibiting properties. For rapid screening of potential DNMT2 binders, a microscale thermophoresis assay was established. Besides the natural inhibitors S-adenosyl-l-homocysteine (SAH) and sinefungin (SFG), we identified new synthetic inhibitors based on the structure of N-adenosyl-2,4-diaminobutyric acid (Dab). Structure-activity relationship studies revealed the amino acid side chain and a Y-shaped substitution pattern at the 4-position of Dab as crucial for DNMT2 inhibition. The most potent inhibitors are alkyne-substituted derivatives, exhibiting similar binding and inhibitory potencies as the natural compounds SAH and SFG. CaCo-2 assays revealed that poor membrane permeabilities of the acids and rapid hydrolysis of an ethylester prodrug might be the reasons for the insufficient activity in cellulo.

Ultra-low-dose computed tomography for torsion measurements of the lower extremities in children and adolescents.

BACKGROUND: Quantifying femoral and tibial torsion is crucial in the preoperative planning for derotation surgery in children and adolescents. The use of an ultra-low-dose computed tomography (CT) protocol might be possible for modern CT scanners and suitable for reliable torsion measurements even though the bones are not completely ossified. METHODS: This is a retrospective review of 77 children/adolescents (mean age 12.7 years) who underwent a lower extremity CT for torsion measurements on a 64-slice scanner. A stepwise dose reduction (70%, 50%, 30% of the original dose) was simulated. Torsion measurements were performed on all image datasets, and image noise, interrater agreement and subjective image quality were evaluated. Effective radiation dose of each original scan was estimated. As proof of concept, 24 children were scanned with an ultra-low-dose protocol, adapted from the 30% dose simulation, and the intra-class correlation coefficient (ICC) was determined. Ethics approval and informed consent were given. RESULTS: Torsion measurements at the simulated 30% dose level had equivalent interrater agreement compared to the 100% dose level (ICC ≥ 0.99 for all locations and dose levels). Image quality of almost all datasets was rated excellent, regardless of dose. The mean sum of the effective dose of the total torsion measurement was reduced by simulation from 0.460/0.490 mSv (boys/girls) at 100% dose to 0.138/0.147 mSv at 30%. The ICC of the proof-of-concept group was as good as that of the simulated 30% dose level. CONCLUSION: Pediatric torsion measurements of the lower extremities can be performed using an ultra-low-dose protocol without compromising diagnostic confidence.

Comparison of temporal evolution of computed tomography imaging features in COVID-19 and influenza infections in a multicenter cohort study.

Purpose: To compare temporal evolution of imaging features of coronavirus disease 2019 (COVID-19) and influenza in computed tomography and evaluate their predictive value for distinction. Methods: In this retrospective, multicenter study 179 CT examinations of 52 COVID-19 and 44 influenza critically ill patients were included. Lung involvement, main pattern (ground glass opacity, crazy paving, consolidation) and additional lung and chest findings were evaluated by two independent observers. Additional findings and clinical data were compared patient-wise. A decision tree analysis was performed to identify imaging features with predictive value in distinguishing both entities. Results: In contrast to influenza patients, lung involvement remains high in COVID-19 patients > 14 days after the diagnosis. The predominant pattern in COVID-19 evolves from ground glass at the beginning to consolidation in later disease. In influenza there is more consolidation at the beginning and overall less ground glass opacity (p = 0.002). Decision tree analysis yielded the following: Earlier in disease course, pleural effusion is a typical feature of influenza (p = 0.007) whereas ground glass opacities indicate COVID-19 (p = 0.04). In later disease, particularly more lung involvement (p < 0.001), but also less pleural (p = 0.005) and pericardial (p = 0.003) effusion favor COVID-19 over influenza. Regardless of time point, less lung involvement (p < 0.001), tree-in-bud (p = 0.002) and pericardial effusion (p = 0.01) make influenza more likely than COVID-19. Conclusions: This study identified differences in temporal evolution of imaging features between COVID-19 and influenza. These findings may help to distinguish both diseases in critically ill patients when laboratory findings are delayed or inconclusive.

Convolutional Neural Networks to Detect Vestibular Schwannomas on Single MRI Slices: A Feasibility Study.

In this study. we aimed to detect vestibular schwannomas (VSs) in individual magnetic resonance imaging (MRI) slices by using a 2D-CNN. A pretrained CNN (ResNet-34) was retrained and internally validated using contrast-enhanced T1-weighted (T1c) MRI slices from one institution. In a second step, the model was externally validated using T1c- and T1-weighted (T1) slices from a different institution. As a substitute, bisected slices were used with and without tumors originating from whole transversal slices that contained part of the unilateral VS. The model predictions were assessed based on the categorical accuracy and confusion matrices. A total of 539, 94, and 74 patients were included for training, internal validation, and external T1c validation, respectively. This resulted in an accuracy of 0.949 (95% CI 0.935-0.963) for the internal validation and 0.912 (95% CI 0.866-0.958) for the external T1c validation. We suggest that 2D-CNNs might be a promising alternative to 2.5-/3D-CNNs for certain tasks thanks to the decreased demand for computational power and the fact that there is no need for segmentations. However, further research is needed on the difference between 2D-CNNs and more complex architectures.

Chemical biology and medicinal chemistry of RNA methyltransferases.

RNA methyltransferases (MTases) are ubiquitous enzymes whose hitherto low profile in medicinal chemistry, contrasts with the surging interest in RNA methylation, the arguably most important aspect of the new field of epitranscriptomics. As MTases become validated as drug targets in all major fields of biomedicine, the development of small molecule compounds as tools and inhibitors is picking up considerable momentum, in academia as well as in biotech. Here we discuss the development of small molecules for two related aspects of chemical biology. Firstly, derivates of the ubiquitous cofactor S-adenosyl-l-methionine (SAM) are being developed as bioconjugation tools for targeted transfer of functional groups and labels to increasingly visible targets. Secondly, SAM-derived compounds are being investigated for their ability to act as inhibitors of RNA MTases. Drug development is moving from derivatives of cosubstrates towards higher generation compounds that may address allosteric sites in addition to the catalytic centre. Progress in assay development and screening techniques from medicinal chemistry have led to recent breakthroughs, e.g. in addressing human enzymes targeted for their role in cancer. Spurred by the current pandemic, new inhibitors against coronaviral MTases have emerged at a spectacular rate, including a repurposed drug which is now in clinical trial.

Diffusion-Weighted Imaging Distinguishes Between Osteomyelitis, Bone Marrow Edema, and Healthy Bone on Forefoot Magnetic Resonance Imaging.

BACKGROUND: Diagnosis of osteomyelitis by imaging can be challenging. The feasibility of diffusion-weighted imaging (DWI) as ancillary sequence was evaluated in this study. PURPOSE: To evaluate DWI for differentiation between osteomyelitis, bone marrow edema, and healthy bone on forefoot magnetic resonance imaging (MRI). STUDY TYPE: Prospective. SUBJECTS: A total of 60 consecutive patients undergoing forefoot MRI divided into three study groups (20 subjects each): osteomyelitis, bone marrow edema, and healthy bone. FIELD STRENGTH/SEQUENCE: A 1.5T and 3T MRI scanners; readout-segmented multishot echo planar DWI. ASSESSMENT: Two independent radiologists measured apparent diffusion coefficient (ADC) values within abnormal or healthy bone. STATISTICAL TESTS: ADC values were compared between groups (pairwise t-test with Bonferroni-Holm correction for multiple testing). Intraclass correlation coefficient (ICC) was calculated to assess inter-reader agreement. Threshold ADC values were determined as the cutoffs that maximized the sum of sensitivity and specificity. Receiver operating characteristic (ROC) analysis was performed with statistical threshold of P < 0.05. RESULTS: Inter-reader agreement was 0.92 in the healthy bone group and 0.78 in both the edema and osteomyelitis groups. Average ADC values were significantly different between groups: 1432 ± 222 × 10-6  mm2 /sec (osteomyelitis), 1071 ± 196 × 10-6  mm2 /sec (bone marrow edema), and 277 ± 89 × 10-6  mm2 /sec (healthy bone). A threshold ADC value of 534 × 10-6  mm2 /sec distinguishes between healthy and abnormal bone with specificity and sensitivity of 100% each. For distinction between osteomyelitis and bone marrow edema, two cutoff values were determined: a 95%-specificity cutoff indicating osteomyelitis (>1320 × 10-6  mm2 /sec) and a 95%-sensitivity cutoff indicating bone marrow edema (<1155 × 10-6  mm2 /sec). Diagnostic accuracy of 95% was achieved for 73% (29/40) of the subjects. DATA CONCLUSION: DWI with ADC maps distinguishes between healthy and abnormal bone on forefoot MRI. Calculated cutoff values allow confirmation or exclusion of osteomyelitis in a high proportion of subjects. EVIDENCE LEVEL: 2 TECHNICAL EFFICACY: Stage 2.

Retrospective evaluation of routine in-hospital observation in 433 patients after CT-guided biopsies.

BACKGROUND: After computed tomography (CT)-guided interventions, routine in-hospital observation is recommended by the Cardiovascular and Interventional Radiological Society of Europe. PURPOSE: To evaluate the frequency of delayed major complications or hospitalizations after CT-guided biopsies in patients with initially no or minor complications and to assess whether routine in-hospital observation is justified. MATERIAL AND METHODS: This retrospective study included 433 outpatients after CT-guided biopsy of the thoracic (n = 176), abdominal (n = 129), or musculoskeletal (n = 128) region with subsequent in-hospital observation. Complications were graded according to the current Society of Interventional Radiology recommendations and grouped into minor or major. A complication that occurred during in-hospital observation was defined as delayed complication. A delayed major complication was a newly developed major complication or a progression from an initially minor to a major complication. Hospitalization frequencies were evaluated similarly. Occurrence, 95% confidence intervals (CI), and P values for significant differences between the three organ groups were calculated. If delayed major complications were more frequent than 1%, routine in-hospital observation was considered justified. RESULTS: Delayed, major complication frequencies were: thoracic, 8.2% (95% CI 4.6-13.4); abdominal, 0.0% (95% CI 0.0-2.9); and musculoskeletal, 0.0% (95% CI 0.0-2.9) (P < 0.001). Delayed hospitalization frequencies were: thoracic, 8.8% (95% CI 5.0-14.2); abdominal, 1.6% (95% CI 0.2-5.6); and musculoskeletal, 0.0% (95% CI 0.0-2.9) (P < 0.001). CONCLUSION: After thoracic interventions, routine observation is considered justified for patient safety whereas routine observation may be omitted after musculoskeletal interventions. In the abdominal group, no delayed complications were observed, but delayed hospitalization occurred. Thus, in-hospital observation could be justified in a safe patient environment, but remains an individual decision.

Short tau inversion recovery (STIR) after intravenous contrast agent administration obscures bone marrow edema-like signal on forefoot MRI.

OBJECTIVE: Short tau or short TI inversion recovery (STIR) MRI sequences are considered a robust fat suppression technique. However, STIR also suppresses signals from other tissues with similar T1 relaxation times. This study investigates the in vivo effect of intravenous gadolinium-based T1-shortening contrast agent on STIR signal. MATERIALS AND METHODS: Institutional board approval and informed consent was obtained. MRI examinations (1.5-T or 3-T) of 31 prospectively included patients were analyzed by two readers. Signal intensity of degenerative bone marrow edema-like signal at the Lisfranc joint on precontrast STIR images and on STIR images acquired after intravenous contrast agent administration (gadoteric acid, gadolinium: 0.5 mmol/ml, 15 ml) was measured. The medial cuneiform bone without observable bone marrow edema-like signal was considered a healthy tissue and served as a reference. Relative changes in signal intensity between precontrast and postcontrast images were calculated for the two tissues. Wilcoxon signed-rank test served for statistical analyses. RESULTS: In bone marrow edema-like signal, both readers observed a median signal change of -35% (interquartile range (IQR) 24) and -34% (IQR 21), respectively, on postcontrast STIR images compared to precontrast STIR. In healthy tissue, the signal remained constant on postcontrast STIR images (median change -2%, IQR 15, and 0%, IQR 17) respectively. For both readers, postcontrast signal change in bone marrow edema-like signal differed from that in healthy tissue (p < 0.001). CONCLUSION: Intravenous gadolinium-based contrast agent causes a significant reduction of signal intensity in bone marrow edema-like signal on routine STIR images. Thus, pathological MRI findings may be obscured.

Convolutional Neural Networks for Classifying Laterality of Vestibular Schwannomas on Single MRI Slices-A Feasibility Study.

Introduction: Many proposed algorithms for tumor detection rely on 2.5/3D convolutional neural networks (CNNs) and the input of segmentations for training. The purpose of this study is therefore to assess the performance of tumor detection on single MRI slices containing vestibular schwannomas (VS) as a computationally inexpensive alternative that does not require the creation of segmentations. Methods: A total of 2992 T1-weighted contrast-enhanced axial slices containing VS from the MRIs of 633 patients were labeled according to tumor location, of which 2538 slices from 539 patients were used for training a CNN (ResNet-34) to classify them according to the side of the tumor as a surrogate for detection and 454 slices from 94 patients were used for internal validation. The model was then externally validated on contrast-enhanced and non-contrast-enhanced slices from a different institution. Categorical accuracy was noted, and the results of the predictions for the validation set are provided with confusion matrices. Results: The model achieved an accuracy of 0.928 (95% CI: 0.869-0.987) on contrast-enhanced slices and 0.795 (95% CI: 0.702-0.888) on non-contrast-enhanced slices from the external validation cohorts. The implementation of Gradient-weighted Class Activation Mapping (Grad-CAM) revealed that the focus of the model was not limited to the contrast-enhancing tumor but to a larger area of the cerebellum and the cerebellopontine angle. Conclusions: Single-slice predictions might constitute a computationally inexpensive alternative to training 2.5/3D-CNNs for certain detection tasks in medical imaging even without the use of segmentations. Head-to-head comparisons between 2D and more sophisticated architectures could help to determine the difference in accuracy, especially for more difficult tasks.

Combined hamartoma of the retina and retinal pigment epithelium - MRI features of a rare paediatric intraocular tumour.

Combined hamartoma of the retina and retinal pigment epithelium is a rare benign ocular tumour in children, composed of glial cells, vascular tissue, and sheets of pigmented epithelial cells. Although generally thought to be congenital, acquired cases are known to exist. It usually presents with reduced visual acuity and/or strabismus and it can be associated with several syndromes, including Neurofibromatosis Type 2. There is no consensus on the management of combined hamartoma of the retina and retinal pigment epithelium. We present a case, including MRI features, of a 4,5-years-old girl with a combined hamartoma of the retina and retinal pigment epithelium.

CT-guided navigated microwave ablation (MWA) of an unfavorable located breast cancer metastasis in liver segment I.

For percutaneous minimally-invasive local ablation therapies of malignant lesions within the liver computed tomography (CT) fluoroscopy or ultrasound (US) can be applied for the positioning of ablation probes. However, lesions in liver segment I and in the upper part of liver segment VIII are difficult to reach with CT fluoroscopy and US guidance even for experienced interventionalists as steep and transcostal access paths may be needed. In addition, there is always the risk to lacerate crucial vessels near the liver hilus. We report on the use of a CT-based stereotactic navigation system (CAS-One, CAScination AG, Bern, Switzerland) for the precise positioning of the ablation probe to perform a percutaneous stereotactic image-guided microwave ablation of a breast cancer liver metastasis in liver segment I that was unreachable with conventional CT or US guidance. Based on the initial planning scan and image-to-patient registration a precise positioning of the probe was possible sparing vital structures like the directly adjacent vulnerable vessels. The ablation was performed without complications fully covering the metastatic lesion with the ablation zone. To this day, there was no recurring tumor 18 months after the intervention.

Discovery of novel dual inhibitors of receptor tyrosine kinases EGFR and PDGFR-ß related to anticancer drug resistance.

With ongoing resistance problems against the marketed EGFR inhibitors having a quinazoline core scaffold there is a need for the development of novel inhibitors having a modified scaffold and, thus, expected lower EGFR resistance problems. An additional problem concerning EGFR inhibitor resistance is an observed heterodimerization of EGFR with PDGFR-ß that neutralises the sole inhibitor activity towards EGFR. We developed novel pyrimido[4,5-b]indoles with varied substitution patterns at the 4-anilino residue to evaluate their EGFR and PDGFR-ß inhibiting properties. We identified dual inhibitors of both EGFR and PDGFR-ß in the nanomolar range which have been initially screened in cancer cell lines to prove a benefit of both EGFR and PDGFR-ß inhibition.

Discovery of novel substituted benzo-anellated 4-benzylamino pyrrolopyrimidines as dual EGFR and VEGFR2 inhibitors.

The quinazoline scaffold is the main part of many marketed EGFR inhibitors. Resistance developments against those inhibitors enforced the search for novel structural lead compounds. We developed novel benzo-anellated 4-benzylamine pyrrolopyrimidines with varied substitution patterns at both the molecular scaffold and the attached residue in the 4-position. The structure-dependent affinities towards EGFR are discussed and first nanomolar derivatives have been identified. Docking studies were carried out for EGFR in order to explore the potential binding mode of the novel inhibitors. As the receptor tyrosine kinase VEGFR2 recently gained an increasing interest as an upregulated signaling kinase in many solid tumors and in tumor metastasis we determined the affinity of our compounds to inhibit VEGFR2. So we identified novel dually acting EGFR and VEGFR2 inhibitors for which first anticancer screening data are reported. Those data indicate a stronger antiproliferative effect of a VEGFR2 inhibition compared to the EGFR inhibition.

The selection and characterization of antibodies to minichromosome maintenance proteins that highlight cervical dysplasia.

BACKGROUND: Screening efforts using the Papanicolaou test have significantly reduced the incidence of cervical cancer in countries with an active screening program. However, this test does not accurately identify all abnormal cases. Significant effort has been expended investigating molecular markers that could improve the sensitivity and specificity of detection of high-grade disease. In this study, we describe the selection and characterization of a set of antibodies to the minichromosome maintenance proteins MCM6 and MCM7 that highlight cervical disease in an immunoassay. METHODS: Antibodies to MCM6 or MCM7 proteins were identified from hybridoma clones screened against tissue microarrays containing different grades of diseased cervical tissue along with normal controls. We determined epitopes by western blotting against nested truncations of either the MCM6 or MCM7 proteins fused to GFP protein. We also determined specificity by western blotting against a panel of major MCM proteins (MCM2-MCM8). Affinity to recombinant antigen and epitope-only peptides was determined using solution-phase binding and determination of free antibody concentration by ELISA. Optimization studies resulted in the selection of antibodies specific to MCM6 and MCM7 for use in immunocytochemistry (ICC) with cervical cytology samples. RESULTS: Four antibodies were identified that demonstrated strong nuclear staining of abnormal cervical epithelial cells in immunohistochemistry (IHC) of cervical biopsies with minimal background staining of normal cervical tissues. Of these four antibody clones, 2E6.7 (MCM7) and 9D4.3 (MCM6) were chosen for further study. Linear epitopes of at most 12 amino acids were identified and verified by binding to epitope-only peptides. Affinities of at least 4×10(-9) M were determined for these two antibodies and both were found to be specific for their respective antigens by western blotting. Clones 9D4.3 and 2E6.7 were also determined to stain abnormal cells in high-grade squamous intraepithelial lesion cytology samples, with minimal background staining of normal cells. CONCLUSION: In this study, we present a method for selecting antibodies that perform well in IHC and ICC applications and characterize two antibodies generated by this method that effectively stain abnormal cells in cervical cancer tissue and cervical cytology samples.

Characterisation of a novel matrix metalloproteinase inhibitor on pancreatic adenocarcinoma cells in vitro and in an orthotopic pancreatic cancer model in vivo.

Matrix metalloproteinases (MMPs) play a central role in tissue maintenance, inflammation and during tumour invasion and metastasis. The impact of MMPs in cancer has encouraged the development of novel MMP-inhibitors without adverse effects on the cell viability. We describe here the synthesis and characterisation of a triazine-derivative as a highly potent MMP-inhibitor. The new compound Triazin 17-2 was developed on the basis of a triazine backbone as a well known and well tolerated chemical scaffold. It was de novo synthesized and tested for MMP inhibition in a cell free assay. In vitro characterisation included tests for cell viability, protein expression and MMP activity in PancTu-1 cells. Effectivity of MMP inhibition was analysed in vitro by invasion assay. Triazin 17-2 was investigated in vivo using an orthotopic pancreatic ductal adenocarcinoma (PDAC) xenograft model in SCID/bg mice. Triazin 17-2 proved to have no adverse effects on cell viability in vitro at concentrations effectively inhibiting MMPs in an invasion assay. Application of Triazin 17-2 in vivo in the orthotopic PDAC model in SCID/bg mice showed a significant reduction of primary tumour weight using conservative therapy and inhibition of metastasis in adjuvant therapy. The MMP-inhibitor Triazin 17-2 was developed and characterised in vitro and in vivo. The new compound has no intrinsic activity to kill cells but is very effective in inhibition of MMPs. In vivo testing revealed that MMP-inhibitors are useful tools in anticancer therapy reducing tumour size and invasion even without direct effects on cell survival.