Vergleichende Untersuchung der Proteinasen zweier Bacillus-Arten (Bacillus subtilis und Bacillus cereus)

Ohne ZusammenfassungTeil der Dissertation, Universität Frankfurt am Main 1958.     

Integrated multi-omics analysis reveals unique signatures of paclitaxel-loaded poly(lactide-co-glycolide) nanoparticles treatment of head and neck cancer cells

The use of poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) as carriers for chemotherapeutic drugs is regarded as an actively targeted nano-therapy for the specific delivery of anti-cancer drugs to target cells. However, the exact mechanism by which PLGA NPs boost anticancer cytotoxicity at the molecular level remains largely unclear. This study employed different molecular approaches to define the response of carcinoma FaDu cells to different types of treatment, specifically: paclitaxel (PTX) alone, drug free PLGA NPs, and PTX-loaded PTX-PLGA NPs. Functional cell assays revealed that PTX-PLGA NPs treated cells had a higher level of apoptosis than PTX alone, whereas the complementary, UHPLC-MS/MS (TIMS-TOF) based multi-omics analyses revealed that PTX-PLGA NPs treatment resulted in increased abundance of proteins associated with tubulin, as well as metabolites such as 5-thymidylic acid, PC(18:1(9Z)/18:1(9Z0), vitamin D, and sphinganine among others. The multi-omics analyses revealed new insights about the molecular mechanisms underlying the action of novel anticancer NP therapies. In particular, PTX-loaded NPs appeared to exacerbate specific changes induced by both PLGA-NPs and PTX as a free drug. Hence, the PTX-PLGA NPs’ molecular mode of action, seen in greater detail, depends on this synergy that ultimately accelerates the apoptotic process, resulting in cancer cell death.     

Identification of Orchidaceae species of Northern West of Syria based on chloroplast DNA

The plant family Orchidaceae has a great economic value (ornamental and medical uses, beside the aromatic features). Traditionally, identification of orchid species has relied heavily on morphological features. These features, however, are either not variable enough between species or too plastic to be used for identification at the species level. DNA-based markers could be the alternative strategy towards an accurate and robust identification of those species. Since the chloroplast DNA has a lower level of evolution compared to the nuclear genome, an attempt was made in this study to investigate polymorphism in the chloroplast DNA among orchid species distributed in North-West region of Syria using Cleaved Amplified Polymorphic Sequence (CAPS) technique for developing markers for the diagnosis of targeted species. CAPS analysis was carried out on 34 orchid samples that represent all species observed in the region. Universal primers were used to amplify targeted chloroplast regions. Generated PCR products were digested with various restriction enzymes. CAPS results revealed high polymorphism among species examined. This polymorphism was suffiecient for the diagnosis of all of those species apart from five species (Ophrys fuciflora (one sample), Oph. bornmuelleri, Ophrys sp., scolopax and Oph. argolica). Availability of such species-specific markers would ensure more authentic identification of orchid species compared to morphological characters and can be regarded as a valuable tool to guide in conservation programs of orchid species in Syria. CAPS data generated were converted to an identification key for orchid species studied.     

Cardiac myofibrillar proteins: Biochemical markers to estimate myocardial injury

Ischaemic heart disease represents the most common of the serious health problems in the contemporary society and acute myocardial infarction (AMI) is the major cause of cardiovascular morbidity and death. The accurate localization and determination of the infarct size and the volume of myocardium at risk at the time of insult is crucial and vital for the choice of treatment. Initially the ischaemic cells are reversibly injured. However, if these changes are not reverted at the earliest, it results in the death of the myocyte. This irreversible myocyte necrosis travels transmurally towards epicardium in the form of a wavefront [1]. A timely intervention during evolving infarct could reduce and delimit the infarct and preserve the left ventricular function [2].Enzyme analysis and electrocardiography (ECG) along with the clinical history of the patient is still considered to constitute a reliable triad in the diagnosis of myocardial infarction (MI) [3]. Efforts have been made to relate infarct size with the serum enzyme level changes without much success. In addition, a number of specialist techniques such as planar radioisotope imaging, single photon emission computed tomography (SPECT), positron emission tomography (PET), Echocardiography, Ventriculography and nuclear magnetic resonance (NMR) imaging have been devised to support diagnosis in the patients who show ambiguous symptoms and ECG findings. However most of these procedures are unavailable to the patients due to economic reasons while others have suffered due to non-availability of ideal radiopharmaceuticals. Major advances have been made in the methods based on immunological techniques to improve the detection and estimation of infarct. These methods are exclusively based upon the production and availability of specific antibodies against intracellular, cardiac specific components [4].     

Enzyme activities in the serum of rainbow trout,Salmo gairdneri Richardson; the effects of water temperature*

Activities of lactate dehydrogenase, hydroxy butyric dehydrogenase, glutamic oxalacetic transaminase, glutamic pyruvic transaminase, glutamate dehydrogenase, creatinine kinase, alkaline phosphatase, and leucine amino peptidase were determined in the sera of rainbow trout. The animals had previously been adapted to temperatures of 3.5, 6, 8, 10, 12.5, 15, 17, 19, 21.5 and 23° C. Most of the enzyme activity increased with the rise in temperature. The activity of alkaline phosphatase decreased in the range 6–19° C, while the changes in the glutamate dehydrogenase activity took a complex course. The results are compared with the findings of other authors.     

Effects of yeast extract on the production of phenylpropanoid metabolites in callus culture of purple basil (Ocimum Basilicum L. var purpurascens) and their in-vitro evaluation for antioxidant potential

Ocimum basilicum L. var. purpurascens is an enriched reservoir of pharmaceutically important compounds with plenty of health and therapeutic attributes such as phenolic acids and anthocyanins. However, the inefficient production of aforementioned metabolites in wild has restricted its commercial utilization. Herein, commercially viable phytochemicals have been enhanced through elicitation of in-vitro cultures of O. basilicum using yeast extract.The impact of various concentrations (YE 1 mg/L,YE 10 mg/L, YE 25 mg/L, YE 50 mg/L, YE 100 mg/L, YE 200 mg/L and YE 400 mg/L) of yeast extract on biomass accumulation, phytochemical production, and antioxidant activities were assessed in callus cultures. Moderate concentration of yeast extract (100 mg/L) enhanced biomass accumulation i.e. fresh weight (FW 216.28 g/L) and dry weight (DW 15.49 g/L) up to 1.5 folds as compared to control (FW 167.14 g/L and DW 10.25 g/L). Similarly, yeast extract (100 mg/L) increased total phenolic and flavonoid contents as well as enhanced antioxidant activities such as ABTS (2,2 azinobis 3-ethylbenzthiazoline-6-sulphonic acid), FRAP (ferric reducing antioxidant power) and DPPH (2,2-diphenyl-1-picryhydrazyl). High performance liquid chromatography (HPLC) analysis was elucidated for further phytochemical investigation. HPLC analysis showed an increase of almost 1.9 folds as compared to control in rosmarinic acid (15.19 mg/g DW), chicoric acid (2.13 mg/g DW), peonidin (2.70 mg/g DW) and cyanidin (1.57 mg/g DW). Likewise, 1.8 fold and 2.4 folds increase was observed in eugenol essential oils (0.25 mg/g DW) and chavicol (0.037 mg/g DW), respectively. For cellular antioxidant activity, reactive oxygen specie or reactive nitogen specie (ROS/RNS) was induced in yeast cells and the effect of O. basilicum callus culture was further investigated in stressed yeast cells. A positive correlation exists between the antioxidant activities, TPC and TFC analysis. In short, these results showed that yeast extract could act as an efficient elicitor to enhance pharmacologically important metabolites in callus cultures of Ocimum basilicum.

Graphical abstract

     

Regeneration of phytoplasma-free <Emphasis Type="Italic">Artemisia roxburghiana</Emphasis> Besser var. purpurascens (Jacq.) Hook. plants using apical meristem culture

We have used apical meristem culture to develop an efficient protocol for reducing phytoplasma infection of Artemisia roxburghiana Besser var. purpurascens (Jacq.) Hook. plants. Shoot tips of different sizes from phytoplasma-infected field-grown plants were treated with 0.1% mercuric chloride (HgCl₂) for 1 min followed by a 30-s exposure to 70% ethanol. The size of the explants significantly influenced the survival frequency and the success of aseptic culture establishment. Sterile explants responded notably to 13.95 μM Kinetin (Kn) and 0.27 μM α-naphthalene acetic acid (NAA), and a maximum of 38 ± 0.87 shoots per explant could be obtained after 6 weeks of incubation. Sub-culturing of the shoot mass after 8 weeks of culture on the previously described medium to 8.88 μM 6-benzyladenine (BA)- and 0.27 μM NAA-containing medium stimulated further multiplication, elongation and growth of each individual regenerant. Efficient rooting was noted after 5 weeks of transfer on half-strength MS medium containing 4.93 μM IBA. Sequential hardening as hydroponic cultures under culture room and glasshouse conditions led to almost 85 and 98% survival of the regenerants upon transfer to pots and field, respectively. Unlike the original A. roxburghiana plants, the plants raised from tissue culture showed a total absence of inherent phytoplasma infection evaluated via inspection of morphological features, PCR and microscopic observations.