Antihormone treatment differentially regulates PSA secretion, PSMA expression and 68Ga-PSMA uptake in LNCaP cells.

BACKGROUND: In recent years, a variety of innovative therapeutics for castration-resistant prostate cancer have been developed, including novel anti-androgenic drugs, such as abiraterone or VPC-13566. Therapeutic monitoring of these pharmaceuticals is performed either by measuring PSA levels in serum or by imaging. PET using PSMA ligands labeled with Fluor-18 or Gallium-68 is the most sensitive and specific imaging modality for detection of metastases in advanced prostate cancer. To date, it remains unclear how PSMA expression is modulated by anti-hormonal treatment and how it correlates with PSA secretion. METHODS: We analyzed modulation of PSMA-mRNA and protein expression, 68Ga-PSMA uptake and regulation of PSA secretion by abiraterone or VPC-13566 in LNCaP cells in vitro. RESULTS: We found that abiraterone and VPC-13566 upregulate PSMA protein and mRNA expression but block PSA secretion in LNCaP cells. Both anti-androgens also enhanced 68Ga-PSMA uptake normalized by the number of cells, whereas abiraterone and VPC-13566 reduced 68Ga-PSMA uptake in total LNCaP monolayers treated due to cell death. CONCLUSION: Our data indicate that PSA secretion and PSMA expression are differentially regulated upon anti-androgen treatment. This finding might be important for the interpretation of 68Ga-PSMA PET images in monitoring therapies with abiraterone and VPC-13566 in prostate cancer patients, but needs to be validated in vivo.

Multicenter validation of cancer gene panel-based next-generation sequencing for translational research and molecular diagnostics.

The simultaneous detection of multiple somatic mutations in the context of molecular diagnostics of cancer is frequently performed by means of amplicon-based targeted next-generation sequencing (NGS). However, only few studies are available comparing multicenter testing of different NGS platforms and gene panels. Therefore, seven partner sites of the German Cancer Consortium (DKTK) performed a multicenter interlaboratory trial for targeted NGS using the same formalin-fixed, paraffin-embedded (FFPE) specimen of molecularly pre-characterized tumors (n = 15; each n = 5 cases of Breast, Lung, and Colon carcinoma) and a colorectal cancer cell line DNA dilution series. Detailed information regarding pre-characterized mutations was not disclosed to the partners. Commercially available and custom-designed cancer gene panels were used for library preparation and subsequent sequencing on several devices of two NGS different platforms. For every case, centrally extracted DNA and FFPE tissue sections for local processing were delivered to each partner site to be sequenced with the commercial gene panel and local bioinformatics. For cancer-specific panel-based sequencing, only centrally extracted DNA was analyzed at seven sequencing sites. Subsequently, local data were compiled and bioinformatics was performed centrally. We were able to demonstrate that all pre-characterized mutations were re-identified correctly, irrespective of NGS platform or gene panel used. However, locally processed FFPE tissue sections disclosed that the DNA extraction method can affect the detection of mutations with a trend in favor of magnetic bead-based DNA extraction methods. In conclusion, targeted NGS is a very robust method for simultaneous detection of various mutations in FFPE tissue specimens if certain pre-analytical conditions are carefully considered.

LEA29Y expression in transgenic neonatal porcine islet-like cluster promotes long-lasting xenograft survival in humanized mice without immunosuppressive therapy.

Genetically engineered pigs are a promising source for islet cell transplantation in type 1 diabetes, but the strong human anti-pig immune response prevents its successful clinical application. Here we studied the efficacy of neonatal porcine islet-like cell clusters (NPICCs) overexpressing LEA29Y, a high-affinity variant of the T cell co-stimulation inhibitor CTLA-4Ig, to engraft and restore normoglycemia after transplantation into streptozotocin-diabetic NOD-SCID IL2rγ-/- (NSG) mice stably reconstituted with a human immune system. Transplantation of INSLEA29Y expressing NPICCs resulted in development of normal glucose tolerance (70.4%) and long-term maintenance of normoglycemia without administration of immunosuppressive drugs. All animals transplanted with wild-type NPICCs remained diabetic. Immunohistological examinations revealed a strong peri- and intragraft infiltration of wild-type NPICCs with human CD45+ immune cells consisting of predominantly CD4+ and CD8+ lymphocytes and some CD68+ macrophages and FoxP3+ regulatory T cells. Significantly less infiltrating lymphocytes and only few macrophages were observed in animals transplanted with INSLEA29Y transgenic NPICCs. This is the first study providing evidence that beta cell-specific LEA29Y expression is effective for NPICC engraftment in the presence of a humanized immune system and it has a long-lasting protective effect on inhibition of human anti-pig xenoimmunity. Our findings may have important implications for the development of a low-toxic protocol for porcine islet transplantation in patients with type 1 diabetes.

[Prognostic value of lacrimal duct diagnostics after tube removal : Retrospective analysis of risk of relapse during the first postoperative year after transcanalicular lacrimal duct surgery with silicone tubing]. / Prognostische Aussagekraft der Tränenwegsdiagnostik nach Intubatentfernung : Retrospektive Analyse des Rezidivrisikos im ersten postoperativen Jahr nach transkanalikulärer Tränenwegschirurgie mit Silikonschlauchintubation.

BACKGROUND: Transcanalicular lacrimal duct surgery has become more important over the past two decades. OBJECTIVES: The aim of the study was to prove the prognostic value of postoperative lacrimal syringing and the testing of spontaneous drainage of lacrimal fluid immediately after tube removal. METHODS: A total of 110 cases with postoperative lacrimal syringing and 183 cases with verification of the postoperative lacrimal fluid drainage performed between January 2001 and August 2008 were retrospectively evaluated. The indication for postoperative diagnostics was set by the investigator. The prognostic value of these two procedures was determined by using prognostic parameters (positive predictive value, PPV; negative predictive value, NPV) and analyzing recurrence nonexistence via Cox regression and Kaplan-Meier estimator. The observation period was limited to 12 months. Predominantly, recurrence was defined on the patient's satisfaction and absence of symptoms, which was determined with the help of a questionnaire. RESULTS: Postoperative verification of the lacrimal syringing is a suitable instrument to estimate surgical success within the first year after lacrimal surgery with a PPV of 92.31%. Testing of the spontaneous drainage of lacrimal fluid after tube removal reached a PPV of 63.33%. CONCLUSION: The proven prognostic value shows that syringing of the lacrimal duct and verification of spontaneous drainage should be integrated into postoperative care in a standardized manner. Hereby, early information for the patient about the expected result of the surgical procedure is enabled.

[Medical and psychological aspects of the treatment of connatal dacryostenosis : Parental evaluation of their own and their child's stress]. / Medizinpsychologische Aspekte bei der Behandlung kindlicher Tränenabflussstörungen : Elterliche Beurteilung eigenen und kindlichen Stresserlebens.

BACKGROUND: Lacrimal probing and syringing for connatal dacryostenosis can be performed under local (LA) and general anesthesia (GA). In cases of invasive medical procedures, pain and anxiety can be distressing for children and their parents. MATERIALS AND METHODS: Using questionnaires (n = 65), parents were asked to evaluate their own stress and that of their child during lacrimal probing and syringing. Analyzing different subgroups, the impact of the kind of anesthesia (LA vs. GA), trust in medical treatment, therapeutic success, prior experiences with GA, parental educational level, age of parents and children, number of children, and time between the intervention and the interview on the stress was examined. Stress level was evaluated on a scale from 1 (no stress) to 10 (maximal stress). RESULTS: Mean children's age was 8.5 ± 7.42 months. Mean age of the parents was 30.8 ± 6.17 years. Treating children under LA, parents reported moderate to severe stress levels for themselves (mean, M = 7.15) and for their children (M = 7.82). Children's stress levels were significantly higher when the treatment was performed under LA (n = 47; M = 7.34) in comparison to GA (n = 18; M = 6.06; p < 0.05). Parents having two or more children reported significantly lower stress levels than those with only one child. Furthermore, prior experiences with GA led to significantly higher parental stress levels when their children were treated under GA. Other factors did not show any impact on parent's and children's stress levels. CONCLUSIONS: Parents might have been influenced by hearing the children's reaction (e. g., crying) during the intervention under LA. Nevertheless, parents had a higher acceptance of this type of intervention (LA) in comparison to GA. This effect was even stronger among parents with prior experience of GA.

[Congenital nasolacrimal duct obstruction from an ophthalmologist's point of view : Causes, diagnosis and staged therapeutic concept]. / Die konnatale Dakryostenose aus ophthalmologischer Sicht : Ursachen, Diagnostik und therapeutisches Stufenkonzept.

BACKGROUND: Congenital nasolacrimal duct obstruction (CNLDO) is one of the most frequent issues in pediatric ophthalmology. There are different therapeutic options, including conservative treatments as well as surgical interventions. These strategies can be classified using a staged therapeutic concept. OBJECTIVE: What are the therapeutic strategies for CNLDO from an ophthalmologist's point of view? METHODS: In this review, different kinds of CNLDO are described. Existing therapeutic interventions are dedicated to the different forms of CNLDO and summarized in a staged therapeutic concept. RESULTS: CNLDO is not a homogeneous disease. Different and also combined pathologies exist. According to symptoms and localization of stenosis, different therapeutic interventions are available. The most common CNLDO manifestation is a persistent Hasner's membrane. If conservative approaches have failed, pressurized probing and syringing of the nasolacrimal duct is the method of choice. This can usually be done under local anesthesia. Since some children show persisting symptoms, other therapies (probing and syringing under general anesthesia with nasolacrimal duct intubation, external dacryocystorhinostomy) have to be considered. Here, dacryoendoscopy offers additional diagnostic and therapeutic options. CONCLUSION: The precise classification of CNLDO and knowledge concerning possible therapeutic interventions are essential. Due to ongoing development of surgical approaches, an increasingly individualized therapy is possible. A gradual therapeutic regimen is available for CNLDO, which has to be adapted individually.

[External Dacryocystorhinostomy--Analysis of Patient Material of the University Hospital Halle from 2000 to 2011]. / Die externe Dakryozystorhinostomie--eine Analyse des Krankenguts der Universitäts-Augenklinik Halle der Jahre 2000-2011.

BACKGROUND: The aim of this retrospective study was to collect additional data on the long-term success (LTS) of external dacryocystorhinostomy (ext-DCR) and the impact of pre-, intra- and postoperative factors on the surgical outcome. This was intended to increase the precision of the indication for DCR. METHOD: A retrospective, non-comparative study was conducted on 637 ext-DCR due to dacryocystitis performed at the Department of Ophthalmology, University Hospital Halle. This included all surgical interventions on patients of at least 11 years of age. Using standardised questionnaires and patient records, 60.75 % (n = 387) of patients were surveyed. Follow-up was 1.0 to 12.0 years (mean, 4.0 years). RESULTS: Analysis of patient satisfaction showed satisfactory (20.2 %) and very satisfactory (74.2 %) results. LTS was 94.4 % (n = 365). Factors negatively influencing postoperative outcome were prior surgical interventions of nose and/or sinus, previous ext-DCR and transcanalicular lacrimal surgery. Surgical outcome was positively influenced by lacrimal sac size and lacrimal stenting. Large saccus lacrimales and use of monocanalicular intubates improved LTS. CONCLUSIONS: LTS of ext-DCR shows very good results, thus underlining its superiority to other surgical approaches. Special attention should be paid to diseases and previous surgical interventions on the nose and/or sinus that effect the lacrimal duct system. Therefore, a special committee of ENT physicians and ophthalmologists has been established at the Department of Ophthalmology, University Hospital Halle. From the surgical point of view, it is important to create an adequate mucosal anastomosis using lacrimal and nasal mucosa. Restoration is possible with lacrimal stent materials. The indication for ext-DCR was restricted by competition with transcanalicular endoscopic interventions to preserve physiological lacrimal drainage.

[Manifestation of bilateral choroidal osteoma in childhood. Progressive myopia due to staphyloma posticum]. / Manifestation eines bilateralen chorioidalen Osteoms im Kindesalter. Progressive Myopie durch Staphyloma posticum.

BACKGROUND: Case report of a 6-year-old boy with bilateral choroidal osteoma. CASE REPORT: The patient was followed up for the next 6 years and during this period bilateral tumor progression was observed. In one eye a staphyloma posticum developed in the area of the osteoma which led to secondary anisometric myopia. CONCLUSION: If undetected, this rare complication can lead to anisometric amblyopia; therefore, repeated cycloplegic refractometry is advisable in children with choroidal osteoma.

[Treatment of congenital lacrimal duct obstruction: A prospective clinical cohort study]. / Behandlung kindlicher Tränenwegsstenosen : Eine prospektive klinische Kohortenstudie.

BACKGROUND: Pressurized probing and irrigation is the method of choice for congenital lacrimal duct obstruction after conservative therapeutic strategies have failed. The aim of this study was to evaluate age dependence and success rates of different therapeutic options. MATERIAL AND METHODS: A prospective cohort study of children with congenital lacrimal duct obstruction was performed. All children treated during the period between May 2009 and June 2010 were included. Surgical success was defined as the absence of epiphora and mucous discharge for a postoperative follow-up of 3 months. Odds ratios (OR) and 95 % confidence intervals (CI) of different variables were estimated using logistic regression. RESULTS: A total of 111 children (137 eyes) underwent treatment for congenital lacrimal duct obstruction. The mean age of the children was 7.7 months (range 1-30 months). The success rate of probing under local anesthesia was 85.5 % (n = 100). Probing under general anesthesia without intubation of the lacrimal pathway was successful in 28.6 % (n = 2). Treatment under general anesthesia with intubation of the lacrimal pathway had a success rate of 94.2 %. If the primary treatment was complicated the OR was 1.9 (95 % CI: 0.7-5.1), for males the OR was 1.7 (95 % CI: 0.8-3.5) and in cases of a positive family history the OR was 1.5 (95 % CI: 0.7-3.3). With every increasing month of age the risk not to be treated successfully increased (OR: 1.3, 95 % CI: 1.2-1.5). CONCLUSION: Syringing under local anesthesia is an effective therapy. The highest success rates were found at the ages of 1-6 months. Therapy with additional silicone tube intubation was highly successful.

[Endoscopy of the Lacrimal Duct System in Children]. / Tränenwegsendoskopie im Kindesalter.

BACKGROUND: Pathologies of the lacrimal duct system show a frequent occurrence in paediatric ophthalmology. Mostly, the connection between the nasolacrimal duct and the nose fails to open but also combined diseases or congenital anomalies may be the reason. Because of complications, the chance for healing after a conservative therapeutic approach decreases and surgical intervention is necessary. PATIENTS AND METHODS: The opportunity for transcanalicular endoscopy of the lacrimal duct system in children is shown by the presentation of three different case reports. Typical clinical findings are given and the use of dacryoendoscopy for diagnostic and therapeutic benefit is pointed out. Therefore, we present an 8-week-old child, suffering recurrent purulent inflammation due to an amniotocele (1), a 5-year-old child having a congenital lacrimal fistula (2) and another 5-year-old child with a severe chronic dacryocystitis of both eyes after several lacrimal duct surgeries showing remaining intrasaccal silicone tubes (3). RESULTS: In all these cases transcanalicular endoscopy could be used successfully for reconstruction of the lacrimal duct systems. A bullous Hasner's membrane could be localized and opened (1). The lacrimal fistula was identified to communicate with the common canaliculus and combined stenosis of the canaliculus and saccus were treated (2). In the third case fragments of intrasaccal silicone tubes could be localised and the foreign bodies could be evacuated by transcanalicular surgery (3). The children with lacrimal fistula and the intrasaccal foreign bodies were treated with self-threading silicone tubing which was removed three months later. In the follow-up period (16 months in case 2, 22 months in case 3 and 38 months in case 1) recurrences of the lacrimal pathologies or clinical complaints were absent. CONCLUSION: Transcanalicular endoscopy of the lacrimal duct system should not be regarded as the means of choice but it does offer additional diagnostic and therapeutic options for special indications. A main advantage of this kind of surgery is its minimally invasive character. Under visual control, topographic anatomy can be preserved. Dacryoendoscopy in children should be done only by experienced surgeons.

[Reconstruction of isolated orbital floor fractures with a prefabricated titanium mesh]. / Versorgung isolierter Orbitabodenfrakturen - Rekonstruktion mit präfabriziertem Titanmesh.

PURPOSE: Concerning safety and efficiency in surgery of isolated orbital floor fractures, several treatment modalities are being discussed. It is a matter of recent interest that prefabricated titanium meshes provide significant advantages. The purpose of this study was to review the usability of the MatrixORBITAL® plate. MATERIAL AND METHODS: In a prospective study we evaluated 10 patients (June 2011 to November 2013) with an isolated orbital floor fracture. All patients in the study group had undergone reconstruction with the MatrixORBITAL® plate. For purposes of analysis were recorded: age, gender, fracture category according to Jaquiéry et al. (2007), duration of surgery, complications, globe motility (Harms wall), exophthalmometry, function of the infraorbital nerve, evaluation of facial symmetry and aesthetic as well as personal contentment of the patient. A reference group received a PDS® sheet, an antral balloon or other materials. RESULTS: In the study group (2 women and 8 men aged between 26 and 83 years) all cases presented a large disruption of the orbital floor (> 2 cm2 bony defect; category II-III). Volume changes were universally associated with diplopia, but only in two cases with an entrapment of the inferior rectus muscle. The mean duration of the operation was 110 minutes (minimum 50, maximum 135 minutes). Postoperatively diplopia was decreasing in all cases. None of the patients had a significant impairment of visus and no reoperation was necessary. The analysis of functional outcomes between the groups mesh, PDS sheet and antral balloon revealed no significant differences. CONCLUSIONS: Based on this experience and due to good intraoperative handling, the authors recommend the prefabricated titanium plate in the treatment of isolated orbital wall fractures or in combination with the medial wall despite the material costs.

Landscape scale estimation of soil carbon stock using 3D modelling.

Soil C is the largest pool of carbon in the terrestrial biosphere, and yet the processes of C accumulation, transformation and loss are poorly accounted for. This, in part, is due to the fact that soil C is not uniformly distributed through the soil depth profile and most current landscape level predictions of C do not adequately account the vertical distribution of soil C. In this study, we apply a method based on simple soil specific depth functions to map the soil C stock in three-dimensions at landscape scale. We used soil C and bulk density data from the Soil Survey for England and Wales to map an area in the West Midlands region of approximately 13,948 km(2). We applied a method which describes the variation through the soil profile and interpolates this across the landscape using well established soil drivers such as relief, land cover and geology. The results indicate that this mapping method can effectively reproduce the observed variation in the soil profiles samples. The mapping results were validated using cross validation and an independent validation. The cross-validation resulted in an R(2) of 36% for soil C and 44% for BULKD. These results are generally in line with previous validated studies. In addition, an independent validation was undertaken, comparing the predictions against the National Soil Inventory (NSI) dataset. The majority of the residuals of this validation are between ± 5% of soil C. This indicates high level of accuracy in replicating topsoil values. In addition, the results were compared to a previous study estimating the carbon stock of the UK. We discuss the implications of our results within the context of soil C loss factors such as erosion and the impact on regional C process models.

[Chronic dacryocystitis in a pediatric patient: transcanalicular endoscopic-assisted removal of iatrogenic foreign body from the lacrimal sac]. / Chronische Dakryozystitis im Kindesalter: Transkanalikuläre endoskopiegestützte Entfernung iatrogener intrasakkaler Fremdkörper.

Congenital dacryostenoses are frequent problems in pediatric ophthalmology. For surgical intervention various transcanalicular techniques by lining the tear ducts with silicone tubes are available. A 5-year-old child was presented with severe chronic dacryocystitis of both eyes after several interventions for lacrimal duct surgery with silicone intubation had been carried out. Using endoscopy the fragments of intrasaccal silicone tubes could be localized and the foreign bodies could be removed by endoscopic transcanicular surgery. The complete removal of silicone tubes is obligatory otherwise severe complications can occur.

Preparation and use of human sclera grafts in ophthalmic surgery.

INTRODUCTION: Human sclera grafts are widely used in ophthalmic surgery. Mainly they are used for coating orbital implants after enucleation. METHODS: For the preparation of sclera grafts, all other tissues must be removed from the donor bulb including the retina, choroid, cornea, corpus vitreum and lens. The sclera graft can be stored dry or in ethanol until transplantation. RESULTS AND CONCLUSION: The processing of sclera grafts in an eye bank is easy to handle compared to the complexity of cornea transplants. The common way is dry storing for at least 1 year. Thus, the demand for sclera grafts can be covered without a lot of trouble.

Connective tissue changes in a mouse model of vein graft disease.

AIM: The extracellular matrix plays an important physiological role in the architecture of the vascular wall. In arterialized vein grafts severe early changes, such as thrombosis and neointimal hyperplasia occur. Paclitaxel is in clinical use as antiproliferative coating of coronary stents. We aimed to investigate the early connective tissue changes in arterialized vein grafts and the influence of perivascular paclitaxel treatment in an in vivo model. METHODS: C57 black mice underwent interposition of the vena cava into the carotid artery. Neointimal hyperplasia, thrombosis, acid mucopolysaccharides (Alcian), collagen fibers (trichrome Masson), elastic fibers, and apoptosis rate (TUNEL) were quantified in paclitaxel treated veins and controls. RESULTS: In both, controls and paclitaxel treated vein grafts acid mucopolysaccharides and elastic fibers were found predominantly in the neointima, whereas collagen fibers were found mainly in the media and adventitia. At 4 weeks postoperatively the neointimal thickness in controls was 52 (13-130) microm, whereas in 0.6 mg/mL l paclitaxel treated veins it was 103 (43-318) microm (P=0.094). At 8 weeks postoperatively paclitaxel treated veins showed a significantly increased neointimal thickness of 136 (87-199) microm compared with 79 (62-146) microm in controls (P=0.032). There was no difference in apoptosis rate between the two groups (P=NS). Even with the lowest concentration of 0.008 mg/mL paclitaxel veins showed a neointimal thickness of 67 (46-205) microm at 4 weeks postoperatively (P=NS vs controls). CONCLUSION: Early vein graft disease is characterised by an accumulation of acid mucopolysaccharides and elastic fibers in the thickened neointima. Paclitaxel treatment increases the neointimal hyperplasia in mouse vein grafts in vivo.

Filter cubes with built-in ultrabright light-emitting diodes as exchangeable excitation light sources in fluorescence microscopy.

The use of ultrabright light-emitting diodes as a potential substitute for conventional excitation light sources in fluorescence microscopy is demonstrated. We integrated ultrabright light-emitting diodes in the filter block of a conventional fluorescence microscope together with a collimating Fresnel lens, a holographic diffuser and emission filters. This setup enabled convenient changes between different excitation light sources and resulted in high excitation efficiencies. Quantitative comparison of image intensities of test samples revealed that light-emitting diodes yielded intensities in the range of a mercury arc lamp depending on the wavelength. The use of ultrabright light-emitting diodes also enabled luminescence lifetime imaging without the need for image intensification.