Modifications of the 5' region of the CASPON tag's mRNA further enhance soluble recombinant protein production in Escherichia coli.

Background: Escherichia coli is one of the most commonly used host organisms for the production of biopharmaceuticals, as it allows for cost-efficient and fast recombinant protein expression. However, challenging proteins are often produced with low titres or as inclusion bodies, and the manufacturing process needs to be developed individually for each protein. Recently, we developed the CASPON technology, a generic fusion tag-based platform process for high-titer soluble expression including a standardized downstream processing and highly specific enzymatic cleavage of the fusion tag.

CASPON platform technology: Ultrafast circularly permuted caspase-2 cleaves tagged fusion proteins before all 20 natural amino acids at the N-terminus.

Fusion protein technologies improve the expression and purification of recombinant proteins, but the removal of the tags involved requires specific proteases. The circularly permuted caspase-2 (cpCasp2) with its specific cleavage site, efficiently generates the untagged protein. While cleavage with cpCasp2 is possible before all 20 proteinogenic amino acids, cleavage before valine, leucine, isoleucine, aspartate and glutamate suffers from slow, and before proline extremely slow, turnover.

Fusion Tag Design Influences Soluble Recombinant Protein Production in .

Fusion protein technologies to facilitate soluble expression, detection, or subsequent affinity purification in are widely used but may also be associated with negative consequences. Although commonly employed solubility tags have a positive influence on titers, their large molecular mass inherently results in stochiometric losses of product yield. Furthermore, the introduction of affinity tags, especially the polyhistidine tag, has been associated with undesirable changes in expression levels.

[Hashimoto thyroiditis, therapeutic options and extrathyroidal options - an up-to-date overview].

Hashimoto's thyroiditis is one of the most common organspecific autoimune diseases and the most frequent cause of hypothyroidism in areas with sufficient iodine supply. Excessively stimulated T cells CD4+ and their differentiated cells are known to play a key role in the pathogenesis. It is currently accepted that on the one hand genetic susceptibility, environmental factors, existential factors (gender difference) play an important role, on the other hand gut and intestinal microbiota seem to contribute to its development too.

Ultrasound criteria for risk stratification of thyroid nodules in the previously iodine deficient area of Austria - a single centre, retrospective analysis.

Background: We aimed to study the validity of six published ultrasound criteria for risk stratification of thyroid nodules in the former severely iodine deficient population of Austria.

Methods: Retrospective, single centre, observer blinded study design. All patients with a history of thyroidectomy due to nodules seen in the centre between 2004 and 2014 with preoperative in-house sonography and documented postoperative histology were analyzed ( = 195).

[Intracranial pressure monitoring in polytrauma patients with traumatic brain injury].

Background: The monitoring of intracranial pressure (ICP) represents a cornerstone in the intensive care of patients with traumatic brain injury (TBI) and the industry provides various technical solutions to this end. Decompressive craniectomy can be an option if conservative measures fail to reduce excessive ICP.

Objective: To examine the pathophysiology of ICP in trauma, the management of polytrauma involving TBI, and the indications for decompressive craniectomy; and to compare the different monitoring systems and their complications.

Preparative crystallization of a single chain antibody using an aqueous two-phase system.

A simultaneous crystallization and aqueous two-phase extraction of a single chain antibody was developed, demonstrating process integration. The process conditions were designed to form an aqueous two-phase system, and to favor crystallization, using sodium sulfate and PEG-2000. At sufficiently high concentrations of PEG, a second phase was generated in which the protein crystallization occurred simultaneously.

High-resolution analysis of alterations in medullary thyroid carcinoma genomes.

Hereditary and sporadic medullary thyroid carcinoma (MTC) are closely associated with RET proto-oncogene mutations. However, the role of additional changes in the tumor genomes remains unclear. Our objective was the identification of chromosomal regions involved in MTC tumorigenesis and to assess their significance by using MTC-derived cell lines.

Urine toxicology screening in Austrian trauma patients: a prospective study.

Introduction: The question as to whether the patient consumed drugs prior to the trauma and which drugs were consumed, is of prime importance for the anesthesia required during surgery. However, many patients are unwilling or unable (including those with multiple trauma or impaired consciousness, or unconscious patients) to answer this question. The purpose of our prospective multicenter study was to collect data about drug consumption in Austria to determine whether drugs are identifiable in the urine of recently injured individuals and to establish the types of drugs consumed.

[Trauma hospital. Knee arthroplasty as primary therapy for proximal tibial fracture].

Ten patients aged 55-85 years with a tibial head fracture AO B3 or C3 were treated primarily by implantation of an endoprosthesis. There were one unilateral, three superficial, and six revision-type prostheses. Follow-up was 6 months to 3 years; two patients were lost to follow-up.

Improved downstream process for the production of plasmid DNA for gene therapy.

Gene therapy and genetic vaccines promise to revolutionize the treatment of inherited and acquired diseases. Since viral vectors are generally associated with numerous disadvantages when applied to humans, the administration of naked DNA, or DNA packed into lipo- or polyplexes emerge as viable alternatives. To satisfy the increasing demand for pharmaceutical grade plasmids we developed a novel economic downstream process which overcomes the bottlenecks of common lab-scale techniques and meets all regulatory requirements.

Continuous matrix assisted refolding of alpha-lactalbumin by ion exchange chromatography with recycling of aggregates combined with ultradiafiltration.

Continuous matrix assisted refolding (MAR) can be achieved on a solid support by using a continuous chromatographic system. Recycling the aggregate fraction, simultaneously formed during a refolding reaction, can further increase the refolding yield. Due to the nature of this reaction, aggregates are the main reason for a refolding yield below stoichiometric conversion.

Matrix assisted refolding of proteins by ion exchange chromatography.

Two different approaches of matrix assisted refolding have been evaluated and compared to conventional refolding by dilution. Bovine alpha-lactalbumin was used for the studies as model protein. It was adsorbed under denaturing conditions on an ion exchange matrix and refolding was completed on the column prior to elution or, depending on the buffer system, in the eluate.

GFAP versus S100B in serum after traumatic brain injury: relationship to brain damage and outcome.

Research indicates that glial fibrillary acidic protein (GFAP), part of the astroglial skeleton, could be a marker of traumatic brain injury (TBI). S100B, an astroglial protein, is an acknowledged marker of TBI. Our goal was to analyze the relationship of GFAP/S100B to brain damage and outcome, and to compare the accuracy of GFAP/S100B for prediction of mortality after TBI.

Glial fibrillary acidic protein in serum after traumatic brain injury and multiple trauma.

Background: This study aimed to determine whether glial fibrillary acidic protein (GFAP) is released after traumatic brain injury (TBI), whether GFAP is related to brain injury severity and outcome after TBI, and whether GFAP is released after multiple trauma without TBI.

Methods: This prospective study enrolled 114 patients who had TBI with or without multiple trauma (n = 101) or multiple trauma without TBI (n = 13), as verified by computerized tomography. Daily GFAP measurement began at admission (<12 hours after trauma) and continued for the duration of intensive care (1-22 days).

The influence of immediate surgical treatment of proximal femoral fractures on mortality and quality of life. Operation within six hours of the fracture versus later than six hours.

We compared the mortality and outcome of 182 patients with proximal fractures of the femur after immediate and delayed surgical treatment. Seventy-nine patients were operated upon within six hours of the fracture (group 1) and 103 patients were operated upon after this period of time (group 2). At six months follow-up, group 1 had a significantly lower mortality rate.

Prevalence of autoimmune thyroiditis and non-immune thyroid disease in multiple sclerosis.

Since multiple sclerosis (MS) and autoimmune thyroiditis (AIT) are presumed to be of autoimmune origin the correlation of these two diseases is of special interest. The aim of this study was to determine whether there are differences in the prevalence of thyroid disease with special emphasis on AIT compared with MS and normal subjects and whether the presence of thyroid disease correlates with disability, disease course, age, and disease duration. 353 consecutive patients with clinically definite MS, without interferon-beta treatment and 308 patients with low back pain or headache were extensively examined for the presence of non-immune or autoimmune thyroid disease.

[Influence of nocturnal surgery on mortality and complications in patients with hip fractures].

Unlabelled: SUBJECT OF INVESTIGATION: The appropriate time of day for surgery of hip fractures and the question of whether surgery should be performed at night are controversial. We therefore investigated the influence of the time of day on mortality and complication rates in surgery for hip fractures.

Method: A total of 170 persons were included in the study with 128 operations being performed during the day and 42 at night, after 9 p.

[[The treatment of severe head-brain injuries in Austria] ].

Introduction: We performed this study in order to assess epidemiology and current practice of treatment of severe traumatic brain injury in Austria. Our survey followed the methods of a study published by J. Ghajar et al in the USA in 1995 and we compared the results to the Brain Trauma Foundation's "Guidelines For The Management Of Severe Head Injury".

[Increased occurrence of autoimmune thyroiditis in patients with chronic rheumatoid arthritis].

The frequent occurrence of both rheumatoid arthritis and autoimmune thyroiditis was already investigated with in part many conflicting results. We investigated a number of 792 patients (383 of them suffering from rheumatoid arthritis and 409 with osteoarthritis). In all patients antithyroid peroxidase and antithyroglobulin antibodies were determined.