Neovii Biotech (Germany)

Landfill leachate and liquid sewage sludge (LS) are two major waste steams generated from solid and liquid waste management processes. Both contain medium chain fatty acids (MCAFs), which are valuable as low-cost feedstocks for bio-based chemicals and lubricant. This study explores the potential of co-fermenting leachate and LS for enhance MCFA production. Two types of leachates (young and old) were used, alongside LS sourced from the decanter tank of a wastewater treatment plant. Leachate and LS were mixed at two ratios (50:50 and 75:25), with ethanol (E) serving as an electron donor under controlled pH and temperature conditions. The results demonstrated that MCFA production was significantly influenced by the age of the leachate and the (leachate: LS) ratio. Notably, the 50:50 mixture showed continuous increases in MCFA production over time. Principal component analysis (PCA) further revealed that young leachate (YL) was the most effective substrate for MCFA elongation, offering the highest acid production and extraction potential. These findings suggest that the co-fermentation of leachate and LS, particularly with young leachate, presents a promising method for MCFA production, supporting waste valorization and sustainable bio-based chemical production. • Producing MCFAs from sludge and leachate is an innovative carbon recovery method. • Using decanter sludge improves MCFA yield and quality with leachate co-substrate. • Leachate age and sludge mix ratios strongly affect chain elongation behavior. • COD and pH tracking give key insights into chain elongation and feedstock status. • Heavy metals and retention time effects need more study for chain elongation.

Kurzzusammenfassung Unterstützt durch rasche technologische Fortschritte haben sich biokatalytische Anwendungen zu einer nachhaltigen, skalierbaren und kostengünstigen Alternative zur etablierten chemischen Katalyse entwickelt. In dieser Arbeit werden die neuesten Beispiele für enzymbasierte Ansätze zur Herstellung von Molekülen mit komplexen Kohlenstoff‐Kohlenstoff‐Gerüsten und mehreren Stereozentren im kommerziellen Maßstab vorgestellt, darunter Peptidbausteine, (seltene) Zucker, synthetische (Oligo‐)Nukleotide und Terpenoide wie Ambrox ® . Neue Enzymklassen und ihre potenziellen Anwendungen – die Synthese von DNA/RNA, die Depolymerisation von synthetischen Kunststoffen oder vollständig enzymatische Schützungs‐/Entschützungsreaktionen – werden hervorgehoben und weisen auf die Vielseitigkeit und breitere industrielle Nutzung biokatalytischer Prozesse hin.

Supported by rapid technological advancements, biocatalytic applications have matured into sustainable, scalable, and cost-competitive alternatives to established chemical catalysis. This review presents the most recent examples of enzyme-based solutions for the manufacturing of molecules with extended carbon-carbon frameworks and multiple stereogenic centers at commercial scale, including peptide building blocks, (rare) sugars, synthetic (oligo)nucleotides, and terpenoids, such as (-)-Ambrox®. Novel enzyme classes are highlighted along with their potential applications-the synthesis of DNA/RNA, the depolymerization of synthetic plastics, or fully enzymatic protection/deprotection schemes-pointing toward the diversification and broader industrial utilization of biocatalysis-based processes.

Androgenetic alopecia (AGA) is the most common cause of hair loss globally, affecting millions of people, particularly men. Current treatments include FDA-approved drugs and devices, but many patients experience side effects or suboptimal results. The niostem device is a new, wearable device that delivers low-level electrical stimulation to promote hair growth. This pilot study aims to evaluate the efficacy and safety of the niostem device in male AGA patients. A total of 21 male patients with AGA used the niostem device daily for 30 min over 6 months. Participants had not used any anti-hair loss products within the 6 months preceding the start of the study. Hair density, thickness, and terminal hair counts were assessed at baseline, 3 months, and 6 months using trichoscopic measurements. Patient-reported outcomes were recorded, and adverse events were monitored. The niostem device resulted in significant increases in hair count, with a 12% increase in total hair density at 3 months and a 19.3% increase at 6 months. Hair thickness also increased by 8.8% in 6 months. Terminal hair density improved significantly over time, with visible hair growth observed in the participants. No adverse events were reported. The niostem device demonstrated a significant increase in hair density and hair thickness in male AGA patients, with no adverse effects. Further large-scale studies are warranted.

Human papillomavirus (HPV) infections are the main cause of cervical and oropharyngeal cancers. As prophylactic vaccines have no curative effect, an efficient therapy would be highly desired. Most therapeutic vaccine candidates target only a small subset of HPV regulatory proteins, namely, E6 and E7, and are therefore restricted in the breadth of their immune response. However, research has suggested E1 and E2 as promising targets to fight HPV+ cancer. Here, we report the design of adenoviral vectors efficiently expressing HPV16 E1 and E2 in addition to transformation-deficient E6 and E7. Vaccination elicited vigorous CD4+ and CD8+ T-cell responses against all encoded HPV16 proteins in outbred mice and against E1 and E7 in C57BL/6 mice. Therapeutic vaccination of C3 tumor-bearing mice led to significantly reduced tumor growth and enhanced survival for both small and established tumors. Tumor biopsies revealed increased numbers of tumor-infiltrating CD8+ T cells in treated mice. Cisplatin enhanced the effect of therapeutic vaccination, accompanied by enhanced infiltration of dendritic cells into the tumor. CD8+ T cells were identified as effector cells in T-cell depletion assays, seemingly under regulation by FoxP3+CD4+ regulatory T cells. Finally, therapeutic vaccination with Ad-Ii-E1E2E6E7 exhibited significantly enhanced survival compared with vaccination with two peptides each harboring a known E6/E7 epitope. We hypothesize that this difference could be due to the induction of additional T-cell responses against E1. These results support the use of this novel vaccine candidate targeting an extended set of antigens (Ad-Ii-E1E2E6E7), in combination with cisplatin, as an advanced strategy to combat HPV+ cancers.

Moss-aGalactosidase A (moss-aGal) is a moss-derived version of human α-galactosidase developed for enzyme replacement therapy in patients with Fabry disease. It exhibits a homogenous N-glycosylation profile with >90% mannose-terminated glycans. In contrast to mammalian cell produced α-galactosidase, moss-aGal does not rely on mannose-6-phosphate receptor mediated endocytosis but targets the mannose receptor for tissue uptake. We conducted a phase 1 clinical trial with moss-aGal in six patients with confirmed diagnosis of Fabry disease during a 28-day schedule. All patients received a single dose of 0.2 mg/kg moss-aGal by i.v.-infusion. Primary endpoints of the trial were safety and pharmacokinetics; secondary endpoints were pharmacodynamics by analyzing urine and plasma Gb3 and lyso-Gb3 concentrations. In all patients, the administered single dose was well tolerated. No safety issues were observed. Pharmacokinetic data revealed a stable nonlinear profile with a short plasma half-life of moss-aGal of 14 minutes. After one single dose of moss-aGal, urinary Gb3 concentrations decreased up to 23% 7 days and up to 60% 28 days post-dose. Plasma concentrations of lyso-Gb3 decreased by 3.8% and of Gb3 by 11% 28 days post-dose. These data reveal that a single dose of moss-aGal was safe, well tolerated, and led to a prolonged reduction of Gb3 excretion. As previously shown, moss-aGal is taken up via the mannose receptor, which is expressed on macrophages but also on endothelial and kidney cells. Thus, these data indicate that moss-aGal may target kidney cells. After these promising results, phase 2/3 clinical trials are in preparation.

Myocardial infarction (MI) causes the loss of cardiac tissue and scar formation, which ultimately lead to heart failure. According to the World Health Organization, heart failure initiated by MI and coronary artery disease accounts for 29% of deaths worldwide. Human Cardiomyocytes in vitro underwent 2 hours of glucose and oxygen deprivation to simulate in vivo myocardial infarction via “Anaerobe Jar” and replacement of cell media with saline. Before this procedure, the treatment group received 200 ul of secretome produced by human adipose derived mesenchymal stem cells, whereas the negative control group received 200 ul of PBS to their 1 ml of culture media. Cell survival was measured by ToxiLight™ BioAssay Kit, a bioluminescent, non-destructive cytolysis assay kit designed to measure the release of the enzyme, adenylate kinase (AK), from damaged cells. Apoptosis and inflammation was measured with ELISA kits for TNF alpha and Nuclear Factor Kappa Beta transcription factor NF-KB. Pre-treatment of cardiomyocytes with ad-MSC secretome before simulating myocardial infarction lead to 23% better survival rate. Pre-treatment of cardiomyocytes with ad-MSC secretome before simulating myocardial infarction lead to reduction of apoptosis (induced cell death, NF-KB) and inflammation (TNFa). In an in vitro model to simulate myocardial infarction, the addition of the secretome of adipose derived mesenchymal stem cells, lead to a better survival rate of cells and a reduction of induced cell death and inflammation. It seems likely, that preventive injections of the secretome of adipose derived mesenchymal stem cells on a regular basis could have the same effect in an anti-ageing / anti-morbidity setting.

Phosphorylcholine (PC)-modified biomolecules like lipopolysaccharides, glycosphingolipids, and (glyco)proteins are widespread, highly relevant antigens of parasites, since this small hapten shows potent immunomodulatory capacity, which allows the establishment of long-lasting infections of the host. Especially for PC-modified proteins, structural data is rare because of the zwitterionic nature of the PC substituent, resulting in low sensitivities and unusual but characteristic fragmentation patterns. We have developed a targeted mass spectrometric approach using hybrid triple quadrupole/linear ion trap (QTRAP) mass spectrometry coupled to nanoflow chromatography for the sensitive detection of PC-modified peptides from complex proteolytic digests, and the localization of the PC-modification within the peptide backbone. In a first step, proteolytic digests are screened using precursor ion scanning for the marker ions of choline (m/z 104.1) and phosphorylcholine (m/z 184.1) to establish the presence of PC-modified peptides. Potential PC-modified precursors are then subjected to a second analysis using multiple reaction monitoring (MRM)-triggered product ion spectra for the identification and site localization of the modified peptides. The approach was first established using synthetic PC-modified synthetic peptides and PC-modified model digests. Following the optimization of key parameters, we then successfully applied the method to the detection of PC-peptides in the background of a proteolytic digest of a whole proteome. This methodological invention will greatly facilitate the detection of PC-substituted biomolecules and their structural analysis.

Background:This is the first study investigating the safety and efficacy of the trifunctional antibody catumaxomab administered i.p. at the end of cytoreductive surgery and postoperatively prior to standard chemotherapy in patients with primary epithelial ovarian cancer (EOC).Methods:Patients received i.p. catumaxomab 10 μg intraoperatively and 10, 20, 50 and 150 μg on days 7, 10, 13 and 16, respectively, postoperatively. After the study, patients received standard chemotherapy and were followed for 23 months. The primary endpoint was the rate of postoperative complications.Results:Forty-one patients entered the study and were evaluable for safety and 34 were alive at 24 months. Complete tumour resection rate was 68%. Postoperative complications were observed in 51%, the most common anastomotic leakage (7%) and wound infections (5%). The most common catumaxomab-related adverse events were abdominal pain, nausea, vomiting and pyrexia. Thirty-nine percent discontinued catumaxomab therapy, and 98% received chemotherapy post study. Kaplan–Meier estimates of disease-free and overall survival after 24 months were 56% and 85%, respectively.Conclusions:Intra- and close postoperative catumaxomab seems feasible, but efficacy and safety were limited by postsurgical complications. In the future prospective trials are needed to investigate the best schedule of integration of catumaxomab into current treatment strategies for EOC.