Identification of the polyketide synthase gene responsible for the synthesis of tanzawaic acids in Penicillium steckii IBWF104-06.

Microorganisms have been used as biological control agents (BCAs) in agriculture for a long time, but their importance has increased dramatically over the last few years. The Penicillium steckii IBWF104-06 strain has presented strong BCA activity in greenhouse experiments performed against phytopathogenic fungi and oomycetes. P. steckii strains generally produce different antifungal tanzawaic acids; interesting compounds known to be catalyzed by polyketide synthetases in other fungi. Since the decalin structure is characteristic for tanzawaic acids, two polyketide synthase genes (PsPKS1 and PsPKS2) were selected for further analysis, which have similarity in sequence and gene cluster structure with genes that are known to be responsible for the biosynthesis of decalin-containing compounds. Subsequently, gene-inactivation mutants of both PsPKS1 and PsPKS2 have been generated. It was found, that the ΔPspks1 mutant cannot produce tanzawaic acids any more, whereas reintegration of the original PsPKS1 gene into the genome of ΔPspks1 reestablished tanzawaic acid production. The mutant ΔPspks2 is not altered in tanzawaic acids production. Interestingly, both mutants ΔPsPKS1 and ΔPsPKS2 still display strong BCA activity, indicating that the mechanism of action is not related to the production of tanzawaic acids.

DBIRD complex integrates alternative mRNA splicing with RNA polymerase II transcript elongation.

Alternative messenger RNA splicing is the main reason that vast mammalian proteomic complexity can be achieved with a limited number of genes. Splicing is physically and functionally coupled to transcription, and is greatly affected by the rate of transcript elongation. As the nascent pre-mRNA emerges from transcribing RNA polymerase II (RNAPII), it is assembled into a messenger ribonucleoprotein (mRNP) particle; this is the functional form of the nascent pre-mRNA and determines the fate of the mature transcript. However, factors that connect the transcribing polymerase with the mRNP particle and help to integrate transcript elongation with mRNA splicing remain unclear. Here we characterize the human interactome of chromatin-associated mRNP particles. This led us to identify deleted in breast cancer 1 (DBC1) and ZNF326 (which we call ZNF-protein interacting with nuclear mRNPs and DBC1 (ZIRD)) as subunits of a novel protein complex--named DBIRD--that binds directly to RNAPII. DBIRD regulates alternative splicing of a large set of exons embedded in (A + T)-rich DNA, and is present at the affected exons. RNA-interference-mediated DBIRD depletion results in region-specific decreases in transcript elongation, particularly across areas encompassing affected exons. Together, these data indicate that the DBIRD complex acts at the interface between mRNP particles and RNAPII, integrating transcript elongation with the regulation of alternative splicing.

P-value-based regulatory motif discovery using positional weight matrices.

To analyze gene regulatory networks, the sequence-dependent DNA/RNA binding affinities of proteins and noncoding RNAs are crucial. Often, these are deduced from sets of sequences enriched in factor binding sites. Two classes of computational approaches exist. The first describe binding motifs by sequence patterns and search the patterns with highest statistical significance for enrichment. The second class uses the more powerful position weight matrices (PWMs). Instead of maximizing the statistical significance of enrichment, they maximize a likelihood. Here we present XXmotif (eXhaustive evaluation of matriX motifs), the first PWM-based motif discovery method that can optimize PWMs by directly minimizing their P-values of enrichment. Optimization requires computing millions of enrichment P-values for thousands of PWMs. For a given PWM, the enrichment P-value is calculated efficiently from the match P-values of all possible motif placements in the input sequences using order statistics. The approach can naturally combine P-values for motif enrichment, conservation, and localization. On ChIP-chip/seq, miRNA knock-down, and coexpression data sets from yeast and metazoans, XXmotif outperformed state-of-the-art tools, both in numbers of correctly identified motifs and in the quality of PWMs. In segmentation modules of D. melanogaster, we detect the known key regulators and several new motifs. In human core promoters, XXmotif reports most previously described and eight novel motifs sharply peaked around the transcription start site, among them an Initiator motif similar to the fly and yeast versions. XXmotif's sensitivity, reliability, and usability will help to leverage the quickly accumulating wealth of functional genomics data.

Routine treatment of patients with chronic lymphocytic leukaemia by office-based haematologists in Germany-data from the Prospective Tumour Registry Lymphatic Neoplasms.

Various treatment options exist for patients with chronic lymphocytic leukaemia (CLL). Clinical registries provide insight into routine treatment and identify changes in treatment over time. The Tumour Registry Lymphatic Neoplasms prospectively collects data on the treatment of patients with lymphoid B-cell neoplasm as administered by office-based haematologists in Germany. Data on patient and tumour characteristics, co-morbidities, systemic treatments, and outcome parameters are recorded. Eight hundred and six patients with CLL were recruited between May 2009 and August 2013. At the start of first-line treatment, median age was 71 years, 64% were male, and 44% had a Binet stage C disease. The most frequently used first-line/second-line regimens were bendamustine + rituximab (BR, 56%/55%), fludarabine + cyclophosphamide + rituximab (FCR, 22%/11%), and bendamustine (B, 5%/9%). Chlorambucil was used in only 7% (first-line) and 6% (second-line) of patients. Patients treated with FCR were younger and healthier than patients treated with BR. Overall, 91% of first-line treatments were successful (40% complete response). Real-life patient populations differ considerably from patients treated in randomized controlled trials. BR and FCR dominate the first-line and second-line treatments of CLL by office-based haematologists in Germany. Future analysis will investigate progression-free and overall survival times.

Twins in spirit part II: DOTATATE and high-affinity DOTATATE--the clinical experience.

PURPOSE: Over recent decades interest in diagnosis and treatment of neuroendocrine tumours (NET) has steadily grown. The basis for diagnosis and therapy of NET with radiolabelled somatostatin (hsst) analogues is the variable overexpression of hsst receptors (hsst1-5 receptors). We hypothesized that radiometal derivatives of DOTA-iodo-Tyr(3)-octreotide analogues might be excellent candidates for somatostatin receptor imaging. We therefore explored the diagnostic potential of (68)Ga-DOTA-iodo-Tyr(3)-octreotate [(68)Ga-DOTA,3-iodo-Tyr(3),Thr(8)]octreotide ((68)Ga-HA-DOTATATE; HA, high-affinity) compared to the established (68)Ga-DOTA-Tyr(3)-octreotate ((68)Ga-DOTATATE) in vivo. METHODS: The study included 23 patients with known somatostatin receptor-positive metastases from NETs, thyroid cancer or glomus tumours who were investigated with both (68)Ga-HA-DOTATATE and (68)Ga-DOTATATE. A patient-based and a lesion-based comparative analysis was carried out of normal tissue distribution and lesion detectability in a qualitative and a semiquantitative manner. RESULTS: (68)Ga-HA-DOTATATE and (68)Ga-DOTATATE showed comparable uptake in the liver (SUVmean 8.9 ± 2.2 vs. 9.3 ± 2.5, n.s.), renal cortex (SUVmean 13.3 ± 3.9 vs. 14.5 ± 3.7, n.s.) and spleen (SUVmean 24.0 ± 6.7 vs. 22.9 ± 7.3, n.s.). A somewhat higher pituitary uptake was found with (68)Ga-HA-DOTATATE (SUVmean 6.3 ± 1.8 vs. 5.4 ± 2.1, p < 0.05). On a lesion-by-lesion basis a total of 344 lesions were detected. (68)Ga-HA-DOTATATE demonstrated 328 lesions (95.3% of total lesions seen), and (68)Ga-DOTATATE demonstrated 332 lesions (96.4%). The mean SUVmax of all lesions was not significantly different between (68)Ga-HA-DOTATATE and (68)Ga-DOTATATE (17.8 ± 11.4 vs. 16.7 ± 10.7, n.s.). CONCLUSION: Our analysis demonstrated very good concordance between (68)Ga-HA-DOTATATE and (68)Ga-DOTATATE PET data. As the availability and use of (68)Ga-HA-DOTATATE is not governed by patent restrictions it may be an attractive alternative to other (68)Ga-labelled hsst analogues.

The XXmotif web server for eXhaustive, weight matriX-based motif discovery in nucleotide sequences.

The discovery of regulatory motifs enriched in sets of DNA or RNA sequences is fundamental to the analysis of a great variety of functional genomics experiments. These motifs usually represent binding sites of proteins or non-coding RNAs, which are best described by position weight matrices (PWMs). We have recently developed XXmotif, a de novo motif discovery method that is able to directly optimize the statistical significance of PWMs. XXmotif can also score conservation and positional clustering of motifs. The XXmotif server provides (i) a list of significantly overrepresented motif PWMs with web logos and E-values; (ii) a graph with color-coded boxes indicating the positions of selected motifs in the input sequences; (iii) a histogram of the overall positional distribution for selected motifs and (iv) a page for each motif with all significant motif occurrences, their P-values for enrichment, conservation and localization, their sequence contexts and coordinates. Free access: http://xxmotif.genzentrum.lmu.de.

The MOF-containing NSL complex associates globally with housekeeping genes, but activates only a defined subset.

The MOF (males absent on the first)-containing NSL (non-specific lethal) complex binds to a subset of active promoters in Drosophila melanogaster and is thought to contribute to proper gene expression. The determinants that target NSL to specific promoters and the circumstances in which the complex engages in regulating transcription are currently unknown. Here, we show that the NSL complex primarily targets active promoters and in particular housekeeping genes, at which it colocalizes with the chromatin remodeler NURF (nucleosome remodeling factor) and the histone methyltransferase Trithorax. However, only a subset of housekeeping genes associated with NSL are actually activated by it. Our analyses reveal that these NSL-activated promoters are depleted of certain insulator binding proteins and are enriched for the core promoter motif 'Ohler 5'. Based on these results, it is possible to predict whether the NSL complex is likely to regulate a particular promoter. We conclude that the regulatory capacity of the NSL complex is highly context-dependent. Activation by the NSL complex requires a particular promoter architecture defined by combinations of chromatin regulators and core promoter motifs.

Investigation of Photodynamic Therapy Promoted by Cherenkov Light Activated Photosensitizers-New Aspects and Revelations.

This work investigates the proposed enhanced efficacy of photodynamic therapy (PDT) by activating photosensitizers (PSs) with Cherenkov light (CL). The approaches of Yoon et al. to test the effect of CL with external radiation were taken up and refined. The results were used to transfer the applied scheme from external radiation therapy to radionuclide therapy in nuclear medicine. Here, the CL for the activation of the PSs (psoralen and trioxsalen) is generated by the ionizing radiation from rhenium-188 (a high-energy beta-emitter, Re-188). In vitro cell survival studies were performed on FaDu, B16 and 4T1 cells. A characterization of the PSs (absorbance measurement and gel electrophoresis) and the CL produced by Re-188 (luminescence measurement) was performed as well as a comparison of clonogenic assays with and without PSs. The methods of Yoon et al. were reproduced with a beam line at our facility to validate their results. In our studies with different concentrations of PS and considering the negative controls without PS, the statements of Yoon et al. regarding the positive effect of CL could not be confirmed. There are slight differences in survival fractions, but they are not significant when considering the differences in the controls. Gel electrophoresis showed a dominance of trioxsalen over psoralen in conclusion of single and double strand breaks in plasmid DNA, suggesting a superiority of trioxsalen as a PS (when irradiated with UVA). In addition, absorption measurements showed that these PSs do not need to be shielded from ambient light during the experiment. An observational test setup for a PDT nuclear medicine approach was found. The CL spectrum of Re-188 was measured. Fluctuating inconclusive results from clonogenic assays were found.

Determination of Radiation Exposure of Individuals in the Population by Patients after Radioiodine Therapy - Comparison of two Measurement Systems. / Ermittlung der Strahlenexposition von Einzelpersonen in der Bevölkerung durch Patienten nach einer Radiojodtherapie ­ Vergleich von 2 Messsystemen.

According to the requirements of radiation protection legislation, patients may only be discharged from the nuclear medicine therapy ward if it is ensured that the cumulative radiation exposure of the population is below 1 mSv per year. In the present study, dose measurements of patients after radioiodine therapy (RIT) and their relatives are to be used to prove that the radiation exposure resulting from the medical application is low and that the legal framework conditions are complied with. Furthermore, the results allow conclusions to be drawn about the measurement accuracy of the dosimeters used. METHODS: In 147 patients after RIT and their relatives, the dosage was measured over 14 days with different measuring systems. Finger ring dosimeters (FRD) were worn during the whole day, furthermore the dose was determined by non-official OSL and TLD dosimeters during the sleep phase. RESULTS: 88 data sets were used for the final analysis. With the FRD, dose values between 0.1-50 mSv were determined for the patients. As expected, the finger ring dose of the relatives was significantly lower, averaging 0.75 mSv compared to 10 mSv for the patient. For the TLD and OSL used in the sleep phase, the measured values were in the same range. The reproducibility of the measurement results was significantly better for the OSL than for the TLD. CONCLUSION: Despite method-related measurement uncertainties, it can be concluded that the exposure dose of patients' relatives after radioiodine therapy is low and that the legal requirements are met. Moreover, the now official OSL dosimeters represent a more accurate and for the chosen measurement task better suited measurement system than the TLD. KEY POINTS: · The exposure dose of patients' relatives after radioiodine therapy is low.. · The requirements of radiation protection legislation after discharge from the nuclear medicine therapy ward are complied with. · OSL dosimeters are a accurate and for the measurement task suited system. CITATION FORMAT: · Hartmann H, Andreeff M, Claußnitzer J et al. Determination of Radiation Exposure of Individuals in the Population by Patients after Radioiodine Therapy - Comparison of two Measurement Systems. Fortschr Röntgenstr 2023; 195: 605 - 612.

Camps 2.0: exploring the sequence and structure space of prokaryotic, eukaryotic, and viral membrane proteins.

Structural bioinformatics of membrane proteins is still in its infancy, and the picture of their fold space is only beginning to emerge. Because only a handful of three-dimensional structures are available, sequence comparison and structure prediction remain the main tools for investigating sequence-structure relationships in membrane protein families. Here we present a comprehensive analysis of the structural families corresponding to α-helical membrane proteins with at least three transmembrane helices. The new version of our CAMPS database (CAMPS 2.0) covers nearly 1300 eukaryotic, prokaryotic, and viral genomes. Using an advanced classification procedure, which is based on high-order hidden Markov models and considers both sequence similarity as well as the number of transmembrane helices and loop lengths, we identified 1353 structurally homogeneous clusters roughly corresponding to membrane protein folds. Only 53 clusters are associated with experimentally determined three-dimensional structures, and for these clusters CAMPS is in reasonable agreement with structure-based classification approaches such as SCOP and CATH. We therefore estimate that ∼1300 structures would need to be determined to provide a sufficient structural coverage of polytopic membrane proteins. CAMPS 2.0 is available at http://webclu.bio.wzw.tum.de/CAMPS2.0/.

Assessing the meaningful change threshold of Quality of Life in Depression Scale using data from two phase 3 studies of esketamine nasal spray.

BACKGROUND: Major depressive disorder (MDD) directly impacts patients' lives including symptoms, functioning and health-related quality-of-life (HRQoL). Patient-reported outcomes can capture these impacts, however interpretation of clinical meaningfulness of these measurements are often not readily available. Meaningful change thresholds (MCTs) can be derived for clinical outcome assessments to quantify the change in symptoms that is meaningful to the patient following pharmacologic treatment or other interventions. The objective of this analysis was to determine the within-patient MCT of the self-reported Quality-of-Life in Depression Scale (QLDS) among patients with MDD and active suicidal ideation with intent (MDSI) using an anchor-based approach. METHODS: Data from 2 randomized phase-3 trials of esketamine nasal spray (ASPIRE I and ASPIRE II) were analyzed. The Montgomery-Åsberg Depression Rating Scale (MADRS) was the primary anchor with three different severity criteria. Other anchor variables utilized were Clinical Global Impression of Severity of Suicidality-revised version, Clinical Global Impression of Imminent Suicide Risk, and EuroQol Visual Analog Scale [EQ-VAS]. Spearman correlation coefficients between the change in QLDS and anchor variables were calculated. The mean change in QLDS score at Day 25 from baseline was calculated based on the categorical change in the anchor. Coefficient yield from linear regression of the mean changes in EQ-VAS and QLDS, and distribution-based approach with ½ SD of change in QLDS were considered. RESULTS: In ASPIRE I, mean (SD) improvement in QLDS score among patients with one category improvement in MADRS from baseline to Day 25 was - 8.22 (8.87), - 8.30 (9.01), and - 8.20 (8.92) using severity criteria #1, #2, and #3, respectively. Patients who achieved a 7-point improvement (MCT) in EQ-VAS yielded a mean - 9.69-point improvement in QLDS at Day 25. The ½ SD of change in QLDS was 5.63. Similar results were obtained for ASPIRE II. The MCTs identified using multiple anchors across both trials ranged from - 11.4 to - 6.7 and had an overall mean of - 7.90 (ASPIRE I) and - 7.92 (ASPIRE II). Thus, an 8-point change was recommended as the MCT for QLDS. CONCLUSION: The recommended MCT will help quantify within-person changes in HRQoL using patient-reported QLDS and determine meaningful treatment benefit in an MDD patient population with acute suicidal ideation or behavior. TRIAL REGISTRATION: Name of the registry: ClinicalTrials.gov. TRIAL REGISTRATION NUMBER: ASPIRE I (NCT03039192), ASPIRE II (NCT03097133). Date of registration: February 01, 2017; March 31, 2017. URL of trial registry record: https://clinicaltrials.gov/ct2/show/NCT03039192 ; https://clinicaltrials.gov/ct2/show/NCT03097133 .

[Treatment Planning and Dose Verification for Combined Internal and External Radiotherapy (CIERT)]. / Bestrahlungsplanung und Dosisverifikation für die kombinierte interne und externe Strahlentherapie (CIERT).

AIM: The combined internal and external radiotherapy (CIERT) take advantage of the benefits from radionuclide therapy and external beam irradiation. These include steep dose gradients and a low toxicity to normal tissue due to the use of unsealed radioisotopes as well as homogeneous dose distribution within the tumor due to external beam irradiation. For a combined irradiation planning, an infrastructure has to be developed that takes into account the dose contributions from both modalities. A physical verification of the absorbed dose distribution should follow by measurements using OSL detectors. METHOD: Internal irradiation was performed using Re-188 in a cylindrical phantom with three inserts. SPECT images were acquired to calculate the internal dose using the software STRATOS. The dose distribution was exported as DICOM-RT data and imported in the software Pinnacle. Based on the internal dose distribution the external irradiation using 6 MV photons was planned. The dose contributions of both modalities separately as well as for combined irradiation was measured using OSL detectors made out of Beryllium oxide. RESULTS: The planed doses of combined irradiation (1 Gy, 2 Gy, 4 Gy) could be verified within the uncertainty of the detectors. The mean energy response to Re-188 was (88.6 ± 2.4) % with respect to the calibration with 200 kV X-ray irradiation. The energy response to 6 MV photons was (146.0 ± 4.9) %. CONCLUSION: A workflow for the treatment planning of combined internal and external radiotherapy has been developed and tested. Measurements verified the calculated doses. Therefore, the physical and technical basis for the dosimetry of combined irradiation were worked out.

Ac-EAZY! Towards GMP-Compliant Module Syntheses of 225Ac-Labeled Peptides for Clinical Application.

The application of 225Ac (half-life T1/2 = 9.92 d) dramatically reduces the activity used for peptide receptor radionuclide therapy by a factor of 1000 in comparison to 90Y, 177Lu or 188Re while maintaining the therapeutic outcome. Additionally, the range of alpha particles of 225Ac and its daughter nuclides in tissue is much lower (47-85 µm for alpha energies Eα = 5.8-8.4 MeV), which results in a very precise dose deposition within the tumor. DOTA-conjugated commercially available peptides used for endoradiotherapy, which can readily be labeled with 177Lu or 90Y, can also accommodate 225Ac. The benefits are lower doses in normal tissue for the patient, dose reduction of the employees and environment and less shielding material. The low availability of 225Ac activity is preventing its application in clinical practice. Overcoming this barrier would open a broad field of 225Ac therapy. Independent which production pathway of 225Ac proves the most feasible, the use of automated synthesis and feasible and reproducible patient doses are needed. The Modular-Lab EAZY is one example of a GMP-compliant system, and the cassettes used for synthesis are small. Therefore, also the waste after the synthesis can be minimized. In this work, two different automated setups with different purification systems are presented. In its final configuration, three masterbatches were performed on the ML EAZY for DOTA-TATE and PSMA-I&T, respectively, fulfilling all quality criteria with final radiochemical yields of 80-90% for the 225Ac-labeled peptides.

Gastroenteropancreatic neuroendocrine tumors: standardizing therapy monitoring with 68Ga-DOTATOC PET/CT using the example of somatostatin receptor radionuclide therapy.

The purpose of this study was to standardize therapy monitoring of hepatic metastases from gastroenteropancreatic neuroendocrine tumors (GEP-NETs) during the course of somatostatin receptor radionuclide therapy (SRRT). In 21 consecutive patients with nonresectable hepatic metastases of GEP-NETs, chromogranin A (CgA) and 68Ga-DOTATOC PET/CT were compared before and after the last SRRT. On 68Ga-DOTATOC PET/CT, the maximum standard uptake values (SUVmax) of normal liver and hepatic metastases were calculated. In addition, the volumes of hepatic metastases (volume of interest [VOI]) were measured using four cut-offs to separate normal liver tissue from metastases (SUVmax of the normal liver plus 10% [VOIliver+10%], 20% [VOIliver+20%], 30% [VOIliver+30%] and SUV  =  10 [VOI10SUV]). The SUVmax of the normal liver was below 10 (7.2 ± 1.3) in all patients and without significant changes. Overall therapy changes (Δ) per patient (mean [95% CI]) were statistically significant with p < .01 for ΔCgA  =  -43 (-69 to -17), ΔSUVmax  =  -22 (-29 to-14), and ΔVOI10SUV  =  -53 (-68 to -38)% and significant with p < .05 for ΔVOIliver+10%  =  -29 (-55 to -3)%, ΔVOIliver+20%  =  -32 (-62 to -2) and ΔVOIliver+30%  =  -37 (-66 to -8). Correlations were found only between ΔCgA and ΔVOI10SUV (r  =  .595; p < .01), ΔSUVmax and ΔVOI10SUV (0.629, p < .01), and SUVmax and ΔSUVmax (r  =  -.446; p < .05). 68Ga-DOTATOC PET/CT allows volumetric therapy monitoring via an SUV-based cut-off separating hepatic metastases from normal liver tissue (10 SUV recommended).

[Radiation Exposition of preparations of radiopharmaceuticals in routinely use, measured with optical-stimulated luminescence (OSL) dosimeters]. / Strahlenexposition bei der Herstellung von Radiopharmaka im nuklearmedizinischen Routinelabor, gemessen mit optisch stimulierten Lumineszenzdetektoren (OSL).

AIM: Because of the new regulation of the radiation protection in Germany in 2018 (Strahlenschutzverordnung) it was meaningful to calculate an overview of the radiation exposition of the personal in the radiopharmaceutical "Hotlab". The regulation demands that the radiation exposition must be minimized, documented and reduced to an optimized level. In the Klinik und Poliklinik für Nuklearmedizin at the University Hospital of Dresden measurements were done with optical-stimulated luminescence (OSL) dosimeters. MATERIALS AND METHODS: Light protected OSL dosimeters were positioned on the fingertips (thumb, forefinger and middle finger) of both hands and on the forehead. Before use the OSL dosimeters were calibrated. The exposition of the hands was measured at three different days in the preparation lab: 99mTc preparation in the daily routine, syringes application for radiosynoviorthesis (RSO) and 90Y Sirtex particles and for preparation of 177Lu and 68Ga radiopharmaceuticals. RESULTS: A relatively high exposition was seen after 99mTc preparations because of the quantity of preparations and syringes. Handling of syringes for RSO lead to an likewise raised exposition. The radiation exposition at preparation of 177Lu and 68Ga radiopharma-ceuticals depends on handling and use of modules. The exposition of the eye lenses was 0.02 mGy or lower and therefore not critical. But an appropriate radiation protection is necessary and a requirement for the most slightly radiation exposition. CONCLUSION: An optimized number of personal is necessary for preparation of radiopharmaceuticals in a "Hotlab" in nuclear medical facility. Therefore, the individual radiation exposition will be limited.

Synthesis and uptake of the compatible solutes ectoine and 5-hydroxyectoine by Streptomyces coelicolor A3(2) in response to salt and heat stresses.

Streptomyces coelicolor A3(2) synthesizes ectoine and 5-hydroxyectoine upon the imposition of either salt (0.5 M NaCl) or heat stress (39 degrees C). The cells produced the highest cellular levels of these compatible solutes when both stress conditions were simultaneously imposed. Protection against either severe salt (1.2 M NaCl) or heat stress (39 degrees C) or a combination of both environmental cues could be accomplished by adding low concentrations (1 mM) of either ectoine or 5-hydroxyectoine to S. coelicolor A3(2) cultures. The best salt and heat stress protection was observed when a mixture of ectoine and 5-hydroxyectoine (0.5 mM each) was provided to the growth medium. Transport assays with radiolabeled ectoine demonstrated that uptake was triggered by either salt or heat stress. The most effective transport and accumulation of [(14)C]ectoine by S. coelicolor A3(2) were achieved when both environmental cues were simultaneously applied. Our results demonstrate that the accumulation of the compatible solutes ectoine and 5-hydroxyectoine allows S. coelicolor A3(2) to fend off the detrimental effects of both high salinity and high temperature on cell physiology. We also characterized the enzyme (EctD) required for the synthesis of 5-hydroxyectoine from ectoine, a hydroxylase of the superfamily of the non-heme-containing iron(II)- and 2-oxoglutarate-dependent dioxygenases (EC 1.14.11). The gene cluster (ectABCD) encoding the enzymes for ectoine and 5-hydroxyectoine biosynthesis can be found in the genome of S. coelicolor A3(2), Streptomyces avermitilis, Streptomyces griseus, Streptomyces scabiei, and Streptomyces chrysomallus, suggesting that these compatible solutes play an important role as stress protectants in the genus Streptomyces.

Molecular imaging using the theranostic agent 197(m)Hg: phantom measurements and Monte Carlo simulations.

BACKGROUND: Radiomercury 197mHg and 197Hg, henceforth referred to as 197(m)Hg, is a promising theranostic radionuclide endowed with properties that allow diagnostic and therapeutic applications. The aim of this work was to investigate the capabilities of 197(m)Hg for nuclear medicine imaging. Therefore measurements were performed by using a Philips BrightView SPECT camera. Furthermore, Monte Carlo simulations using the GATE software were performed to theoretically explore the imaging contribution from the various gamma and X-ray emissions from 197(m)Hg for a commercial clinical camera with low-energy high-resolution (LEHR) and high-energy general-purpose (HEGP) collimators. We estimated the spatial resolution by using a four-quadrant bar phantom, and we evaluated the planar and tomographic images from an abdominal phantom containing three cylindrical sources of 197(m)Hg solution. RESULTS: A good accordance between measurements and simulations was found for planar and SPECT imaging. Simulations allowed the decomposition of the detected energy spectrum into photon origins. Measurements and simulations for the bar phantom revealed that for the LEHR collimator, the 6-mm pattern could be resolved, whereas for the HEGP collimator, the resolution is about 10 mm. Furthermore, we found that no significant image distortion results from high-energy photons when using the LEHR collimator. CONCLUSIONS: We demonstrated the imaging capabilities of 197(m)Hg which is essential both for diagnostic applications and to determine the in vivo biodistribution for dose calculations in therapeutic applications.

Twins in spirit part IV - [177Lu] high affinity DOTATATE. A promising new tracer for peptide receptor radiotherapy?

AIM: Besides the use of somatostatin analogues, small molecules like sunitinib and everolimus as well as conventional chemotherapy, peptide receptor radiotherapy (PRRT) using radiolabelled somatostatin analogues has gained an important role in the treatment of inoperable, metastasized neuroendocrine tumours (NET). There are various radiotracers in use. Based on our experience with the PET tracer [68Ga]DOTA-3-iodo-Tyr3-octreotate ([68Ga]HA-DOTATATE), a DOTATATE derivative with an increased binding affinity to hsst5, the current retrospective analysis is exploring the therapeutic potential of [177Lu]HA-DOTATATE. METHODS: Eighteen patients with metastatic NET (G1/G2) were treated using [177Lu]DOTATATE and/or [177Lu]HA-DOTATATE, and dosimetric results of both tracers were compared. RESULTS: Using [177Lu]HA-DOTATATE, a mean tumour dose of 5.34 Gy/GBq (median 2.53 Gy/GBq; range 0.89-33.3 Gy/GBq) was achieved, while [177Lu]DOTATATE delivered a tumour dose of 5.53 Gy/GBq (median 2.70 Gy/GBq; range 0.44-15.3 Gy/GBq). Organ doses for [177Lu]HA-DOTATATE vs. [177Lu]DOTATATE were as follows: kidney 2.31 ± 0.85 vs. 2.03 ± 0.96 Gy/GBq, liver 1.06 ± 0.79 vs. 1.67 ± 1.73 Gy/GBq, spleen 3.89 ± 4.04 vs. 4.50 ± 3.69 Gy/GBq and whole body 0.16 ± 0.10 Gy/GBq vs. 0.15 ± 0.08 Gy/GBq. Tumour-to-kidney dose ratio was slightly higher for [177Lu]DOTATATE (2.4 ± 5.6) compared to [177Lu]HA-DOTATATE (1.5 ± 3.6). CONCLUSION: Both tracers showed marked inter-patient variation in their dosimetry, and no significant differences in dosimetry of [177Lu]HA-DOTATATE and [177Lu]DOTATATE were observed when taking all patients into account. Thus, [177Lu]HA-DOTATATE appears viable for PRRT, although it was marginally inferior regarding kidney dose and tumour-to-kidney dose ratio compared to the established [177Lu]DOTATATE.