A blueprint of ectoine metabolism from the genome of the industrial producer Halomonas elongata DSM 2581T

The halophilic γ-proteobacterium Halomonas elongata DSM 2581T thrives at high salinity by synthesizing and accumulating the compatible solute ectoine. Ectoine levels are highly regulated according to external salt levels but the overall picture of its metabolism and control is not well understood. Apart from its critical role in cell adaptation to halophilic environments, ectoine can be used as a stabilizer for enzymes and as a cell protectant in skin and health care applications and is thus produced annually on a scale of tons in an industrial process using H. elongata as producer strain. This paper presents the complete genome sequence of H. elongata (4 061 296 bp) and includes experiments and analysis identifying and characterizing the entire ectoine metabolism, including a newly discovered pathway for ectoine degradation and its cyclic connection to ectoine synthesis. The degradation of ectoine (doe) proceeds via hydrolysis of ectoine (DoeA) to Nα-acetyl-l-2,4-diaminobutyric acid, followed by deacetylation to diaminobutyric acid (DoeB). In H. elongata, diaminobutyric acid can either flow off to aspartate or re-enter the ectoine synthesis pathway, forming a cycle of ectoine synthesis and degradation. Genome comparison revealed that the ectoine degradation pathway exists predominantly in non-halophilic bacteria unable to synthesize ectoine. Based on the resulting genetic and biochemical data, a metabolic flux model of ectoine metabolism was derived that can be used to understand the way H. elongata survives under varying salt stresses and that provides a basis for a model-driven improvement of industrial ectoine production

Adjuvant Therapy Using Mistletoe Containing Drugs Boosts the T-Cell-Mediated Killing of Glioma Cells and Prolongs the Survival of Glioma Bearing Mice

Viscum album L. extracts (VE) are applied as complementary cancer therapeutics for more than one century. Extracts contain several compounds like mistletoe lectins (ML) 1-3 and viscotoxins, but also several minor ingredients. Since ML-1 has been described as one of the main active components harboring antitumor activity, purified native or recombinant ML-1 has been also used in clinical trials in the last years. The present study examined and compared the immunoboosting effects of three ML-1 containing drugs (the extract ISCADOR Qu, the recombinant ML-1 Aviscumine, and purified native ML-1) in the context of the T-cell mediated killing of glioma cells. Additionally we examined the possible underlying T-cell stimulating mechanisms. Using cocultures of immune and glioma cells, a PCR-based microarray, quantitative RT-PCR, and an antibody-based array to measure cytokines in blood serum, immunosupporting effects were determined. A highly aggressive, orthotopic, immunocompetent syngeneic mouse glioma model was used to determine the survival of mice treated with ISCADOR Qu alone or in combination with tumor irradiation and temozolomide (TMZ). Treatment of glioblastoma (GBM) cells with ISCADOR Qu that contains a high ML concentration, but also viscotoxins and other compounds, as well as with Aviscumine or native ML-1, enhanced the expansion of cancer cell-specific T-cells as well as T-cell-mediated tumor cell lysis, but to a different degree. In GBM cells all three ML-1-containing preparations modulated the expression of immune response associated genes. In vivo, subcutaneous ISCADOR Qu injections at increasing concentration induced cytokine release in immunocompetent VM/Dk-mice. Finally, ISCADOR Qu, if applied in combination with tumor irradiation and TMZ, further prolonged the survival of glioma mice. Our findings indicate that ML-1 containing drugs enhance anti-GBM immune responses and work in synergy with radiochemotherapy. Therefore, adjuvant mistletoe therapy should be considered as an auspicious treatment option for glioma patients

3D-based buccal augmentation for ideal prosthetic implant alignment-an optimized method and report on 7 cases with pronounced buccal concavities

Objectives Screw-retained restoration of implants is advantageous for biological and esthetic reasons. Due to buccal concavities, however, this preferred type of restoration can only be used in about half of the anterior indications. Based on case series, an optimized method for the treatment of such indications is to be described; the clinical reliability is to be ascertained by means of measurements (before and after augmentation) and assigned to the current literature. Material and methods A case series of seven cases with buccal concavities of the anterior alveolar ridge were treated with optimized method, which is presented step-by-step until the prosthetic restoration. The depths of the bone concavities were measured and related to the bone gain after augmentation procedure respectively after implantation. Results Linear measurements of the buccal concavities showed an average undercut of 4 mm [SD +/- 1.13]. After healing period of six months, the buccal concavities could be compensated bony to such an extent that implants could be inserted in correct position and angulation. On average, there was a horizontal bone gain of 3.7 mm [SD +/- 0.59]. Even after implantation and another six months of healing, stable bone dimensions could be assumed with an average of 4.3 [SD +/- 0.83] mm of bone gain compared to baseline. In six of the seven cases, the favorite screw-retained, one-piece full-ceramic restoration could be fixed on the implants. Due to the implant axis, one case had to be treated with a cemented two-part full-ceramic system. Conclusions With the described optimized method the most favorable screw-retained restoration can also be used in situations with unfavorable concavities of buccal bone. Especially for this indication, a special form of the horizontal deficit, the customized bone regeneration with titanium meshes is highly reliable in terms of healing and extent of augmentation. However, long-term results and a study/control group are required to evaluate the effectiveness of the presented protocol. Clinical relevance. Since these situations require an augmentation that is up to 5 mm thick and a procedure that is as minimally invasive as possible appears to be necessary in the visible area, an optimized method is described in this publication

A Volumetric and Morphological Analysis of Recurrent Odontogenic Keratocysts by Semiautomatic Segmentation

Purpose: The authors conducted this study to provide morphological and volumetric data of recurrent odontogenic keratocysts of the upper and lower jaw to emphasize risk factors in accordance with their radiological appearance and guide clinical decisions for jeopardized patients. Methods: By applying the open-source software ITK-Snap on cone-beam computed tomography images, volumetric measurements of histopathologically diagnosed recurrent odontogenic keratocysts could be performed. For statistical investigations, descriptive statistics and independent Student t test were performed. The intraclass correlation coefficient was used to assess intra- and inter-rater reliabilities. P values P < 0.05 were considered significant. Results: Forty patients (24 male and 16 female) were included in this study. Recurrent odontogenic keratocysts had a mean maximum diameter of 28.91 mm +/- 12.00 mm and a mean volume of 4.48 cm(3) +/- 4.29 cm(3). According to morphology, irregular shape (P = 0.001; P = 0.005), unclear margin (P = 0.001; P = 0.001), multilocular morphology (P = 0.001; P = 0.001), and cortical bone exceedance (P = 0.001; P = 0.007) are statistically significantly associated with a larger cyst diameter and volume. Furthermore, significant differences by diameter and volume could be shown between patients with and without iliac crest graft reconstruction (P = 0.001; P = 0.001). Conclusions: Volumetric analysis reveals that recurrent odontogenic keratocysts show large diametric and volumetric extension that leads to complex reconstruction by iliac crest grafts, adding an argument that special attention should be paid to this entity and its recurrence. in case of difficult histopathological examination, lesions with irregular shape and margin, multilocular morphology, cortical bone exceedance, and clinically visible symptoms should be considered for close morphological and volumetric clinico- radiological follow-up

Anatomical and volumetric analysis of fibro-osseous lesions of the craniofacial skeleton

Purpose: Our study aimed to provide volumetric data relating to fibro-osseous lesions of the craniofacial skeleton, in order to highlight risk factors due to the different entities, and to guide clinical decisions for jeopardized patients. Methods: Volumetric measurements of osteomas and ossifying fibromas were performed by applying the open-source software ITK-Snap to cone-beam computed tomography images. DICOM datasets were imported, identified, and delineated using semiautomatic segmentation; this was then verified using manual segmentation. The volumes of the lesions were computed automatically in cubic millimeters using the program. For statistical investigations, descriptive statistics and independent Student t-tests were performed. Additionally, Pearson's correlation was applied as a bivariate analysis. Values of p < 0.05 were considered significant. Results: 45 patients (11 male and 34 female) were included in this study. The mean volumes were 10.02 +/- 18.79 cm(3) for osteomas and 4.80 +/- 5.71 cm(3) for ossifying fibromas (p = 0.016). Males (12.81 +/- 20.38 cm(3)) presented significantly larger volumes than females (5.43 +/- 10.32 cm(3)) (p = 0.042). With regard to shape, morphology, and affection of surrounding anatomical structures, irregular shape (p = 0.001; p = 0.037), multilocular morphology (p = 0.001; p = 0.037), nerve affection (p = 0.001; p = 0.002), tooth affection (p = 0.001; p = 0.594), cortical bone exceedance (p = 0.033; p = 0.001), and clinically visible symptoms (p = 0.004; p = 0.001) were statistically significantly associated with a larger volume of both entities. Conclusion: Volumetric analysis revealed that osteomas significantly exceeded the mean size of ossifying fibromas, supporting the argument that special attention should be paid to this entity. In cases of difficult histopathological examination, lesions with irregular shape, multilocular morphology, nerve and tooth affection, cortical bone exceedance, and clinically visible symptoms should be considered for close clinico-radiological follow-up, irrespective of the entity. (C) 2021 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved

Phase I trial of r viscumin (INN : aviscumine) given subcutaneously in patients with advanced cancer: A study of the European Organisation for Research and Treatment of Cancer (EORTC protocol number 13001)

Safety of aviscumine by subcutaneous route was assessed in patients with advanced cancer refractory to chemotherapy. Patients with progressive disease received escalating doses twice weekly Treatment of the accrued 26 patients (10 colorectal cancer (CRC), 6 soft tissue sarcoma (STS), 5 melanoma (MM), 5 others) was well tolerated without substance-related grade 3 or 4 toxicities. Grade 1/2 toxicities were predominantly injection site reactions. Aviscumine lacked dose-limiting toxicity (DLT) up to a maximal dose of 10 ng/kg. An increase of interleukin-1 beta and interferon-gamma from baseline was seen in the patient's plasma between the 1st and 11th injection. Highest release of both cytokines was in the dose range of 45.9 ng/kg. Interferon-gamma was not detected after doses higher than 6 ng/kg. Eight patients (5 CRC, 1 MM, 1 STS, 1 RCC) had disease stabilisation for 79-250 days (median122 days) associated with an increase of interleukin (IL)-1 beta and interferon (IFN)-gamma. Aviscumine was well tolerated and appeared to possess clinical activity at a biologically active dose between 4 and 6 ng/kg. (C) 2008 Elsevier Ltd. All rights reserved