The Chemical Composition of Endotoxin Isolated from Intestinal Strain of Desulfovibrio desulfuricans

Desulfovibrio desulfuricans anaerobes are constituents of human alimentary tract microflora. There are suggestions that they take part in the pathogenesis of periodontitis and some gastrointestinal inflammatory disorders, such as ulcerative colitis or Crohn's disease. Endotoxin is one of Gram-negative bacteria cellular components that influence these microorganisms pathogenicity. Endotoxin is a lipid-polisaccharide heteropolymer consisting of three elements: lipid A, core oligosaccharide, and O-specific polysaccharide, also called antigen-O. The biological activity of lipopolysaccharide (LPS) is determined by its structure. In this study, we show that rhamnose, fucose, mannose, glucose, galactose, heptose, and 2-keto-3-deoxyoctulosonic acid (Kdo) are constituents of D. desulfuricans endotoxin oligosaccharide core and O-antigen. Lipid A of these bacteria LPS is composed of glucosamine disaccharide substituted by 3-acyloxyacyl residues: ester-bound 3-(dodecanoyloxy)tetradecanoic, 3-(hexadecanoyloxy)tetradecanoic acid, and amide-bound 3-(tetradecanoyloxy)tetradecanoic acid

Chemical composition of Desulfovibrio desulfuricans lipid A

Lipopolysaccharides also called endotoxins are an integral component of the outer membrane of Gram-negative bacteria. When released from the bacterial surface, they interact with a host immune system, triggering excessive inflammatory response. Lipid A is the biologically most active part of endotoxin, and its activity is modulated by the quantity, quality and arrangement of its fatty acids. Desulfovibrio desulfuricans is sulfate-reducing, Gram-negative bacterium that is supposed to be opportunistic pathogens of humans and animals. In the present study, chemical composition of lipid A from various strains of D. desulfuricans was analyzed by gas chromatography/mass spectrometry. It was found that the fatty acid component of the lipid A contains dodecanoic, tetradecanoic, 3-hydroxytetradecanoic and hexadecanoic acids, and its carbohydrate core is composed of glucosamine. The analysis of 3-acyloxyacyl residue of the lipid A revealed the presence of amide-bound 3-(dodecanoyloxy)tetradecanoic and 3-(hexadecanoyloxy)tetradecanoic acids and ester-bound 3-(tetradecanoyloxy)tetradecanoic acid. It was concluded that both fatty acid and 3-acyloxyacyl residue profiles of the lipid A from the studied bacteria were similar to those of E. coli and S.enterica

Utilization of host iron sources by bacteria Desulfovibrio desulfuricans

Pathogenicity of Gram-negative bacteria is determined by their ability of iron uptake from environmental and human reserve sources. To acquire this element microorganisms synthesize siderophores, iron chelating structures that allow them to utilize various host iron sources such as hemoglobin, myoglobin, ferritin, transferrin and lactoferrin. Host iron sources utilized by Desulfovibrio desulfuricans (D. desulfuricans) are still unrecognized. These microorganisms colonize a human alimentary tract as a component of the natural intestinal microfl ora. However, their involvement in the pathogenesis of intestinal disorders, such as Crohn’s disease or ulcerative colitis, cannot be excluded. The purpose of this study was to analyze the ability of these bacteria to utilize several body iron sources. The aim of the study was realized by the evaluation of number of colonies of pre-starved D. desulfuricans strains after 48 hour culturing on pyruvate Postgate’s medium supplemented with 1.5 mg/dm3 of the iron source (human hemoglobin and transferrin, bovine hemoglobin, transferrin, lactoferrin and hemin, equine myoglobin and cytochrome c). The control cells were cultured on medium devoid of iron. Most of the tested strains D. desulfuricans (except for DV/B) utilized iron from a wide variety host sources. The interstrain diversity of bacterial growth in the presence of each of iron sources was observed. Soil strain DSM 642 was the slowest proliferating one on medium containing both human and bovine transferrin. Therefore, this strain does not utilize iron from both iron sources. The most intensive growth was observed with DV/I and DV/I/1 intestinal strains on medium supplemented with equine myoglobin and cytochrome c, and bovine lactoferrin, whereas DV/H strain proliferated the most on medium containing both human and bovine hemoglobin.Patogenność bakterii Gram-ujemnych jest uwarunkowana ich zdolnością pozyskiwania żelaza ze środowiska oraz rezerw zainfekowanego makroorganizmu. W tym celu bakterie syntetyzują siderofory, czyli układy chelatujące żelazo, które umożliwiają im wykorzystywanie jego hemowych i niehemowych źródeł ustrojowych, takich jak hemoglobina, mioglobina, ferrytyna, transferyna i laktoferyna. Dotychczas nie poznano ustrojowych źródeł żelaza wykorzystywanych przez bakterie Desulfovibrio desulfuricans (D. desulfuricans). Gatunek ten kolonizuje m.in. przewód pokarmowy człowieka, stanowiąc składnik fi zjologicznej mikrofl ory jelita. Nie wyklucza się jednak udziału tych bakterii w etiopatogenezie niektórych schorzeń tego narządu, takich jak choroba Crohna czy wrzodziejące zapalenie jelita grubego. Celem podjętych badań była analiza możliwości wykorzystywania przez bakterie D. desulfuricans różnych ustrojowych źródeł żelaza. Zamierzenie to realizowano oceniając po 48 godzinach hodowli liczbę kolonii wygłodzonych izolatów D. desulfuricans na pirogronianowej pożywce Postgate’a wzbogaconej o 1,5 mg/dm3 określonego źródła żelaza (hemoglobiny i transferyny ludzkiej, hemoglobiny, transferyny, laktoferyny i heminy bydlęcej, mioglobiny i cytochromu c końskiego). Hodowlę kontrolną stanowiły bakterie namnażane się na podłożu z obniżoną ilością żelaza. Większość izolatów D. desulfuricans (oprócz DV/B) pozyskiwała żelazo z różnych źródeł ustrojowych, przy czym stwierdzono ich międzyszczepowe zróżnicowanie wzrostu w obecności każdej z żelazoprotein i heminy. Najwolniej namnażał się glebowy izolat DSM 642 na podłożu zawierającym transferynę ludzką i bydlęcą, nie wykorzystując żelaza zawartego w tym ustrojowym źródle. Wśród wszystkich badanych bakterii najintensywniej namnażały się dzikie szczepy DV/I i DV/I/1 na podłożach z końską mioglobiną i cytochromem c oraz laktoferyną bydlęcą oraz DV/H w obecności ludzkiej i bydlęcej hemoglobiny

Bioresorbable Nonwoven Patches as Taxane Delivery Systems for Prostate Cancer Treatment

Prostate cancer is the second most common cancer in males. In the case of locally advanced prostate cancer radical prostatectomy is one of the first-line therapy. However, recurrence after resection of the tumor can appear. Drug-eluting bioresorbable implants acting locally in the area of the tumor or the resection margins, that reduce the risk of recurrence would be advantageous. Electrospinning offers many benefits in terms of local delivery so fiber-forming polyesters and polyestercarbonates which are suitable to be drug-loaded were used in the study to obtain CTX or DTX-loaded electrospun patches for local delivery. After a fast verification step, patches based on the blend of poly(glycolide-ε-caprolactone) and poly(lactide-glycolide) as well as patches obtained with poly(lactide-glycolide- ε-caprolactone) were chosen for long-term study. After three months, 60% of the drug was released from (PGCL/PLGA) + CTX and it was selected for final, anticancer activity analysis with the use of PC-3 and DU145 cells to establish its therapeutic potential. CTX-loaded patches reduced cell growth to 53% and 31% respectively, as compared to drug-free patches. Extracts from drug-free patches showed excellent biocompatibility with the PC-3 cell line. Cabazitaxel-loaded bioresorbable patches are a promising drug delivery system for prostate cancer therapy

Antiproliferative and Cytotoxic Properties of Propynoyl Betulin Derivatives against Human Ovarian Cancer Cells: In Vitro Studies

Due to the incidence of ovarian cancer (OC) and the limitations of available therapeutic strategies, it is necessary to search for novel therapeutic solutions. The aim of this study was to evaluate the cytotoxic effect of betulin 1 and its propynoyl derivatives 2–6 against ovarian cancer cells (SK-OV-3, OVCAR-3) and normal myofibroblasts (18Co). Paclitaxel was used as the reference compound. The propynoyl derivatives 2–6 exhibited stronger antiproliferative and cytotoxic activities compared to betulin 1. In both ovarian cancer cell lines, the most potent compound was 28-propynoylbetulin 2. In the case of compound 2, the calculated IC50 values were 0.2 µM for the SK-OV-3 cells and 0.19 µM for the OVCAR-3 cells. Under the same culture conditions, the calculated IC50 values for compound 6 were 0.26 µM and 0.59 µM, respectively. It was observed that cells treated with compounds 2 and 6 caused a decrease in the potential of the mitochondrial membrane and a significant change in cell morphology. Betulin 1, a diol from the group of pentacyclic triterpenes, has a confirmed wide spectrum of biological effects, including a significant anticancer effect. It is characterized by low bioavailability, which can be improved by introducing changes to its structure. The results showed that chemical modifications of betulin 1 only at position C-28 with the propynoyl group (compound 2) and additionally at position C-3 with the phosphate group (compound 3) or at C-29 with the phosphonate group (compound 6) allowed us to obtain compounds with greater cytotoxic activity than their parent compounds, which could be used to develop novel therapeutic systems effective in the treatment of ovarian cancer

Anticancer Activity of the Acetylenic Derivative of Betulin Phosphate Involves Induction of Necrotic-Like Death in Breast Cancer Cells In Vitro

Betulin (BT) is a natural pentacyclic lupane-type triterpene exhibiting anticancer activity. Betulin derivatives bearing propynoyloxy and phosphate groups were prepared in an effort to improve the availability and efficacy of the drug. In this study, a comparative assessment of the in vitro anticancer activity of betulin and its four derivatives was carried out using two human breast cancer cell lines: SK-BR-3 and MCF-7. In both studied cell lines, 30-diethoxyphosphoryl-28-propynoylbetulin (compound 4) turned out to be the most powerful inhibitor of growth and inducer of cellular death. Detailed examination of that derivative pertained to the mechanisms underlying its anticancer action. Treatment with compound 4 decreased DNA synthesis and up-regulated p21WAF1/Cip1 mRNA and protein levels in both cell lines. On the other hand, that derivative caused a significant increase in cell death, as evidenced by increased lactate dehydrogenase (LDH) release and ethidium homodimer uptake. Shortly after the compound addition, an increased generation of reactive oxygen species and loss of mitochondrial membrane potential were detected. The activation of caspase-3 and fragmentation of genomic DNA suggested an apoptotic type of cell death. However, analysis of cellular morphology did not reveal any nuclear features typical of apoptosis. Despite necrosis-like morphology, dead cells exhibited activation of the cascade of caspases. These observations have led to the conclusion that compound 4 pushed cells to undergo a form of necrotic-like regulated cell demise

Anticancer Activity of the Acetylenic Derivative of Betulin Phosphate Involves Induction of Necrotic-Like Death in Breast Cancer Cells In Vitro

Betulin (BT) is a natural pentacyclic lupane-type triterpene exhibiting anticancer activity. Betulin derivatives bearing propynoyloxy and phosphate groups were prepared in an effort to improve the availability and efficacy of the drug. In this study, a comparative assessment of the in vitro anticancer activity of betulin and its four derivatives was carried out using two human breast cancer cell lines: SK-BR-3 and MCF-7. In both studied cell lines, 30-diethoxyphosphoryl-28-propynoylbetulin (compound 4) turned out to be the most powerful inhibitor of growth and inducer of cellular death. Detailed examination of that derivative pertained to the mechanisms underlying its anticancer action. Treatment with compound 4 decreased DNA synthesis and up-regulated p21WAF1/Cip1 mRNA and protein levels in both cell lines. On the other hand, that derivative caused a significant increase in cell death, as evidenced by increased lactate dehydrogenase (LDH) release and ethidium homodimer uptake. Shortly after the compound addition, an increased generation of reactive oxygen species and loss of mitochondrial membrane potential were detected. The activation of caspase-3 and fragmentation of genomic DNA suggested an apoptotic type of cell death. However, analysis of cellular morphology did not reveal any nuclear features typical of apoptosis. Despite necrosis-like morphology, dead cells exhibited activation of the cascade of caspases. These observations have led to the conclusion that compound 4 pushed cells to undergo a form of necrotic-like regulated cell demise