New Expression Method and Characterization of Recombinant Human Granulocyte Colony Stimulating Factor in a Stable Protein Formulation

Human recombinant granulocyte colony stimulating factor (rhG-CSF) is widely used in hematology and oncology for the treatment of neutropenia, for the restoration of neutrophil production after bone marrow transplantation, for myelodysplastic syndromes, and aplastic anemia. The E. coli expression system is commonly used for fast recombinant production of rhG-CSF at a large scale. We have applied a novel autoinduction method for the batch expression of rhG-CSF to study whether this new system would increase cell mass and target-protein yield compared to conventional E. coli cell culture and induction with isopropyl ?-D-thiogalactopyranoside (IPTG). We could demonstrate 3-fold higher culture densities and a 5-fold higher protein yield compared to IPTG induction without the need to monitor cell growth in a shortened 24 h expression procedure. rhG-CSF expressed in autoinduction media was successfully extracted from E. coli inclusion bodies and refolded by dialysis. After size exclusion chromatography (SEC) purification, rhG-CSF showed similar conformation, biological activity and aggregation profile compared to the commercially available biosimilar TEVAgrastim® (TEVA Pharma AG). Expression by autoinduction is suggested as a cost- and time-effective method for rhG-CSF production

Applying Quantum Cascade Laser Spectroscopy in Plasma Diagnostics

The considerably higher power and wider frequency coverage available from quantum cascade lasers (QCLs) in comparison to lead salt diode lasers has led to substantial advances when QCLs are used in pure and applied infrared spectroscopy. Furthermore, they can be used in both pulsed and continuous wave (cw) operation, opening up new possibilities in quantitative time resolved applications in plasmas both in the laboratory and in industry as shown in this article. However, in order to determine absolute concentrations accurately using pulsed QCLs, careful attention has to be paid to features like power saturation phenomena. Hence, we begin with a discussion of the non-linear effects which must be considered when using short or long pulse mode operation. More recently, cw QCLs have been introduced which have the advantage of higher power, better spectral resolution and lower fluctuations in light intensity compared to pulsed devices. They have proved particularly useful in sensing applications in plasmas when very low concentrations have to be monitored. Finally, the use of cw external cavity QCLs (EC-QCLs) for multi species detection is described, using a diagnostics study of a methane/nitrogen plasma as an example. The wide frequency coverage of this type of QCL laser, which is significantly broader than from a distributed feedback QCL (DFB-QCL), is a substantial advantage for multi species detection. Therefore, cw EC-QCLs are state of the art devices and have enormous potential for future plasma diagnostic studies

Fine-mapping and identification of candidate causal genes for tail length in the Merinolandschaf breed

Docking the tails of lambs in long-tailed sheep breeds is a common practice worldwide. But this practice is associated with pain. Breeding for a shorter tail could offer an alternative. Therefore, this study aimed to analyze the natural tail length variation in the Merinolandschaf and to identify causal alleles for the short tail phenotype segregating within long-tailed breeds. We used SNP-based association analysis and haplotype-based mapping in 362 genotyped (Illumina OvineSNP50) and phenotyped Merinolandschaf lambs. Genome-wide significant regions were capture sequenced in 48 lambs and comparatively analyzed in various long and short-tailed sheep breeds and wild sheep subspecies. Here we show a SNP located in the first exon of HOXB13 and a SINE element located in the promotor of HOXB13 as promising candidates. These results enable more precise breeding towards shorter tails, improve animal welfare by amplification of ancestral alleles and contribute to a better understanding of differential embryonic development

Splenic rupture and fungal endocarditis in a pediatric patient with invasive fusariosis after allogeneic hematopoietic stem cell transplantation for aplastic anemia: A case report

Background Invasive mold infections are a well-known and life-threatening condition after allogeneic hematopoietic stem cell transplantation (HSCT). While Aspergillus species are recognized as predominant pathogens, Fusarium species should also be considered due to their broad environmental distribution and the expected poor outcome of invasive fusariosis. Particularly, splenic rupture as a complication of disseminated disease has not been reported yet. Case presentation Two weeks after allogeneic HSCT for severe aplastic anemia, a 16-year-old boy presented with painful, erythematous skin nodules affecting the entire integument. As disseminated mycosis was considered, treatment with liposomal amphotericin B and voriconazole (VCZ) was initiated. Invasive fusariosis was diagnosed after histological and previously unpublished polymerase chain reaction-based examination of skin biopsies. Microbiological tests revealed Fusarium solani species. Despite stable neutrophil engraftment and uninterrupted treatment with VCZ, he developed mold disease-associated splenic rupture with hypovolemic shock and fungal endocarditis. The latter induced a cardiac thrombus and subsequent embolic cerebral infarctions with unilateral hemiparesis. Following cardiac surgery, the patient did not regain consciousness because of diffuse cerebral ischemia, and he died on day +92 after HSCT. Conclusion Invasive fusariosis in immunocompromised patients is a life-threatening condition. Despite antimycotic treatment adapted to antifungal susceptibility testing, the patient reported here developed uncommon manifestations such as splenic rupture and fungal endocarditis

High-performance GaAs/Alas terahertz quantum-cascade lasers for spectroscopic applications

We have developed terahertz (THz) quantum-cascade lasers (QCLs) based on GaAs/AlAs heterostructures for application-defined emission frequencies between 3.4 and 5.0 THz. Due to their narrow line width and rather large intrinsic tuning range, these THz QCLs can be used as local oscillators in airborne or satellite-based astronomical instruments or as radiation sources for high-resolution absorption spectroscopy, which is expected to allow for a quantitative determination of the density of atoms and ions in plasma processes. The GaAs/AlAs THz QCLs can be operated in mechanical cryocoolers and even in miniature cryocoolers due to the comparatively high wall-plug efficiency of around 0.2% and typical current densities below 500 A/cm$^2$. These lasers emit output powers of more than 1 mW at operating temperatures up to about 70 K, which is sufficient for most of the abovementioned applications. © 2011-2012 IEEE