Effects Of Aminoglycoside Antibiotics Research Articles

Cytotoxic effect of aminoglycoside antibiotics on the mammalian cell lines

Aminoglycoside antibiotics have been used for treating serious but also routine infections in veterinary and human medicine for many years. The basic aim of this work is to evaluate the cytotoxicity of dihydrostreptomycin and neomycin in vitro on three cell cultures - BHK-21 (Syrian golden hamster kidney fibroblast), VERO (African green monkey kidney fibroblast) and FEA (feline embryonic fibroblast) cells. The morphological changes were examined by Giemsa staining. Cells were dried and visualized under fluorescence microscope. After the exposure to different experimental doses of dihydrostreptomycin (812.5-20000 µg/mL) and neomycin (1000-20000 µg/mL) during 24 h, the viability of BHK-21, FEA and VERO cell lines were evaluated by MTT assay. Viability of BHK-21 cells significantly (P < 0.001) decreased after treatment with 3500; 5500 and 7500 µg/mL of dihydrostreptomycin and 9000; 10000 and 20000 µg/mL of neomycin. The FEA cell viability decreased significantly (P < 0.001; P < 0.01) at 2500 and 3000 µg/mL dihydrostreptomycin and at 3000 µg/mL of neomycin treatment. Only the highest concentration of dihydrostreptomycin (20000 µg/mL) reduced VERO cell viability significantly (P < 0.01). Based on or results we can assume the effect of different antibiotics in different concentrations on cell lines is various. Detection of antibiotic toxicity to animal cells is very important because of the increasing resistance of bacteria. One of the solutions is drug dose increasing, but only to a certain concentration, since the toxic effect over the therapeutic one will prevail, which we have also shown in this work.

ANTIOXIDANT POTENTIAL OF ADEMETIONINE IN GENTAMICINE NEPHROPATHY PROGRESSION

Acute kidney injury has been globally considered as a serious medical issue. Its incidence rate is increasing over the years despite the introduction of new therapies, the cause of which is often a side effect to drugs, or due to the impact of toxins, trauma. Nephrotoxicity is one of the most important side effects of aminoglycoside antibiotics, and on gentamicin, in particular. Despite rigorous patient monitoring, this type of nephrotoxicity appears in 10–30% of therapeutic courses. Therefore, searching for new therapies, which would be able to prevent or decrease toxic kidney injury during the treatment with aminoglycosides is quite relevant. Ademetionine is an amino acid derivative, which possesses various effects such as antioxidant, membrane stabilizing, anti-inflammatory and regenerative.&#x0D; Aim of the research is to study an antioxidative activity of ademetionine in conditions of gentamicin nephropathy progression in rats. Materials and methods. Research was conducted on non-linear mature white rats weighting 130-180 g, randomly divided into 3 groups (n = 7): I group included intact control animals, II group included rodents with gentamicin nephropathy (injection of 4% Gentamicin sulphate solution in a dosage of 80 mg/kg for 6 days), III group involved the rats receiving ademetionine («Abbott SpA», Italy) in a dosage of 20 mg/kg. Peroxidation processes in blood and kidneys were evaluated by the content of active products that react with thiobarbituric acid and oxidative modification of proteins levels, antioxidant defence was assessed by catalase and glutathione peroxidase activity, SH-groups and ceruloplasmin content.&#x0D; Results. In the course of experimental studies on the model of gentamicin nephropathy, the expressive antioxidant activity of ademetionine was proven: it was demonstrated by reducing the intensity of lipid peroxidation (a decrease in the content of active products that react with thiobarbituric acid) and proteins (a decrease in the content of oxidative modified proteins) in the blood and kidney homogenate, next with increased activity of the enzymatic link of antioxidant protection (glutathione peroxidase and catalase) and non-enzymatic (ceruloplasmin and compounds with SH-groups). The antioxidant potential of ademetionine has been confirmed by an increase in the antioxidant-prooxidant index in kidney tissue and a significant decrease in the index of oxidative stress in the blood of treated animals.&#x0D; Conclusion. Thus, the results of experimental studies indicate the antioxidative effect of ademetionine in modelled gentamicin nephropathy, and point out the necessity of further studying its potential nephroprotectective properties.

Partial Aminoglycoside Lesions in Vestibular Epithelia Reveal Broad Sensory Dysfunction Associated with Modest Hair Cell Loss and Afferent Calyx Retraction.

Although the effects of aminoglycoside antibiotics on hair cells have been investigated for decades, their influences on the dendrites of primary afferent neurons have not been widely studied. This is undoubtedly due to the difficulty in disassociating pathology to dendritic processes from that resulting from loss of the presynaptic hair cell. This was overcome in the present investigation through development of a preparation using Chinchilla laniger that enabled direct perilymphatic infusion. Through this strategy we unmasked gentamicin’s potential effects on afferent calyces. The pathophysiology of the vestibular neuroepithelia after post-administration durations of 0.5 through 6 months was assessed using single-neuron electrophysiology, immunohistochemistry, and confocal microscopy. Hair cell densities within cristae central zones (0.5-, 1-, 2-, and 6-months) and utricle peri- and extrastriola (6-months) regions were determined, and damage to calretinin-immunoreactive calyces was quantified. Gentamicin-induced hair cell loss exhibited a profile that reflected elimination of a most-sensitive group by 0.5-months post-administration (18.2%), followed by loss of a second group (20.6%) over the subsequent 5.5 months. The total hair cell loss with this gentamicin dose (approximately 38.8%) was less than the estimated fraction of type I hair cells in the chinchilla’s crista central zone (approximately 60%), indicating that viable type I hair cells remained. Extensive lesions to afferent calyces were observed at 0.5-months, though stimulus-evoked modulation was intact at this post-administration time. Widespread compromise to calyx morphology and severe attenuation of stimulus-evoked afferent discharge modulation was found at 1 month post-administration, a condition that persisted in preparations examined through the 6-month post-administration interval. Spontaneous discharge was robust at all post-administration intervals. All calretinin-positive calyces had retracted at 2 and 6 months post-administration. We found no evidence of morphologic or physiologic recovery. These results indicate that gentamicin-induced partial lesions to vestibular epithelia include hair cell loss (ostensibly reflecting an apoptotic effect) that is far less extensive than the compromise to stimulus-evoked afferent discharge modulation and retraction of afferent calyces (reflecting non-apoptotic effects). Additionally, calyx retraction cannot be completely accounted for by loss of type I hair cells, supporting the possibility for direct action of gentamicin on the afferent dendrite.

Effects of Aminoglycoside Antibiotics on Human Embryonic Stem Cell Viability during Differentiation In Vitro.

Human embryonic stem cells (hESCs) are being used extensively in array of studies to understand different mechanisms such as early human embryogenesis, drug toxicity testing, disease modeling, and cell replacement therapy. The protocols for the directed differentiation of hESCs towards specific cell types often require long-term cell cultures. To avoid bacterial contamination, these protocols include addition of antibiotics such as pen-strep and gentamicin. Although aminoglycosides, streptomycin, and gentamicin have been shown to cause cytotoxicity in various animal models, the effect of these antibiotics on hESCs is not clear. In this study, we found that antibiotics, pen-strep, and gentamicin did not affect hESC cell viability or expression of pluripotency markers. However, during directed differentiation towards neural and hepatic fate, significant cell death was noted through the activation of caspase cascade. Also, the expression of neural progenitor markers Pax6, Emx2, Otx2, and Pou3f2 was significantly reduced suggesting that gentamicin may adversely affect early embryonic neurogenesis whereas no effect was seen on the expression of endoderm or hepatic markers during differentiation. Our results suggest that the use of antibiotics in cell culture media for the maintenance and differentiation of hESCs needs thorough investigation before use to avoid erroneous results.

The effect of aminoglycoside antibiotics on the induction of adventitious shoots in soybean

The effect of aminoglycoside antibiotics on the induction of adventitious shoots in soybean

Assessing Specific Oligonucleotides and Small Molecule Antibiotics for the Ability to Inhibit the CRD-BP-CD44 RNA Interaction

Studies on Coding Region Determinant-Binding Protein (CRD-BP) and its orthologs have confirmed their functional role in mRNA stability and localization. CRD-BP is present in extremely low levels in normal adult tissues, but it is over-expressed in many types of aggressive human cancers and in neonatal tissues. Although the exact role of CRD-BP in tumour progression is unclear, cumulative evidence suggests that its ability to physically associate with target mRNAs is an important criterion for its oncogenic role. CRD-BP has high affinity for the 3′UTR of the oncogenic CD44 mRNA and depletion of CRD-BP in cells led to destabilization of CD44 mRNA, decreased CD44 expression, reduced adhesion and disruption of invadopodia formation. Here, we further characterize the CRD-BP-CD44 RNA interaction and assess specific antisense oligonucleotides and small molecule antibiotics for their ability to inhibit the CRD-BP-CD44 RNA interaction. CRD-BP has a high affinity for binding to CD44 RNA nts 2862–3055 with a Kd of 645 nM. Out of ten antisense oligonucleotides spanning nts 2862–3055, only three antisense oligonucleotides (DD4, DD7 and DD10) were effective in competing with CRD-BP for binding to 32P-labeled CD44 RNA. The potency of DD4, DD7 and DD10 in inhibiting the CRD-BP-CD44 RNA interaction in vitro correlated with their ability to specifically reduce the steady-state level of CD44 mRNA in cells. The aminoglycoside antibiotics neomycin, paramomycin, kanamycin and streptomycin effectively inhibited the CRD-BP-CD44 RNA interaction in vitro. Assessing the potential inhibitory effect of aminoglycoside antibiotics including neomycin on the CRD-BP-CD44 mRNA interaction in cells proved difficult, likely due to their propensity to non-specifically bind nucleic acids. Our results have important implications for future studies in finding small molecules and nucleic acid-based inhibitors that interfere with protein-RNA interactions.

The Effects of Aminoglycoside Antibiotics on Platelet Aggregation and Blood Coagulation

To investigate the effects of different aminoglycoside antibiotics on platelet aggregation and blood coagulation, as well as the underlying mechanisms. Blood samples were collected and prepared as platelet-rich plasma and platelet-poor plasma samples. Then assigned into different groups for the following antibiotics treatments: gentamicin, streptomycin, etimicin, amikacin, and kanamycin, as group 0 mg/L, group 30 mg/L, group 91mg/L, and group 910 mg/L for each drugs. The maximum platelet aggregation rate induced by adenosine diphosphate, expression levels of CD62p and FIB-R, prothrombin time, activated partial thromboplastin time, thrombin time, and fibrinogen were measured. The sodium citrate and sodium heparin were used in whole blood tests for the whole blood coagulation time as well as the Ca(2+) in blood plasma. Amikacin and gentamicin could inhibit the aggregation of platelets, which contributed to the whole blood clotting disorder. Amikacin and gentamicin might inhibit the platelet aggregation by blocking the activation and release of FIB-R or probably the inhibition of endogenous clotting factor as well. This effect was not dependent on calcium ions.

Effect of aminoglycoside antibiotics on the conformation of the unpaired loop adenine of avian leucosis virus RNA as revealed using 2-aminopurine fluorescence

The fluorescent properties of 2-aminopurine (2-AP) incorporated in an RNA sequence are used to study the structural dynamics and local changes of the retroviral RNA structure. Using 2-AP, the conformational states of the unpaired loop adenine in avian leucosis virus RNA were studied upon its interaction with aminoglycoside antibiotics. The intensity of 2-AP fluorescence in the monomeric RNA hairpin was higher than in both RNA dimers. The intensity of fluorescence in the extended dimer was significantly lower than in the kissing loop dimer. The finding was be explained by the fact that stacking contacts in the extended dimer produce a more compact loop structure than in the kissing loop dimer. When the binding of aminogycoside antibiotics with the kissing loop dimer RNA was analyzed, only tobramycin increased the intensity of 2-AP fluorescence almost threefold. The results showed that 2-AP fluorescence is suitable for detecting local changes in complexes of retroviral RNA with ligands.

The effect of aminoglycoside antibiotics on the adventitious regeneration from apricot leaves and selection of nptII-transformed leaf tissues

The effect of different concentrations of the aminoglycoside antibiotics, geneticin, paromomycin and streptomycin on adventitious regeneration from leaf explants of apricot was tested to design an alternative procedure for selecting transgenic shoots. Streptomycin and paromomycin reduced shoot regeneration percentage with increasing concentration of antibiotics. Almost a complete inhibition of regeneration was reached when 20μM paromomycin was used, although up to 40 μM streptomycin was necessary to completely inhibit regeneration. Geneticin had a very toxic effect on apricot leaves and regeneration was inhibited at almost all concentrations tested. Addition of kanamycin hastened the development of adventitious buds although silver thiosulfate and not kanamycin was responsible for the observed increase in the consistency of the results from independent experiments. Kanamycin and paromomycin at the concentrations tested improved selection of transformed cells and resulted in a larger number of gfp-expressing regions. Paromomycin at 40 and 25.7 μM kanamycin improved proliferation of transformed tissues as compared with the other antibiotics used and non-selected controls.

The effect of aminoglycoside antibiotics on acetylcholine-evoked K+ currents in guinea-pig outer hair cells

The effect of aminoglycoside antibiotics on acetylcholine-evoked K+ currents in guinea-pig outer hair cells

The effect of aminoglycoside antibiotics on acetylcholine-evoked K+ currents in guinea-pig outer hair cells

The effect of aminoglycoside antibiotics on acetylcholine-evoked K+ currents in guinea-pig outer hair cells

Some properties of Plesiomonas shigelloides treated with aminoglycosides.

The effect of aminoglycoside antibiotics (amikacin, gentamicin, netilmicin and tobramycin) at sublethal concentrations (sub-MICs) on some properties of Plesiomonas shigelloides strains was evaluated. All agents decreased the bacterial surface hydrophobicity. Amikacin (1/4 of the MIC) and netilmicin (1/4 and 1/8 of the MIC) changed the hydrophobic character of P. shigelloides surface to a hydrophilic one. Treatment of the strains with aminoglycosides decreased also motility, netilmicin being the most effective. No significant changes were found in lipolytic activity of antibiotic-treated strains. In the majority of cases aminoglycosides increased sensitivity of bacteria to hydrogen peroxide. The tested antibiotics did not induce production of short-chained N-acylhomoserine lactones signal molecules. Aminoglycosides at sub-MICs affected important activities of P. shigelloides potentially associated with their virulence in dependence on strain, antibiotic and concentration.

Effect of aminoglycoside antibiotics on in-vitro morphogenesis from cultured cells of chrysanthemum and tobacco

Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro expiants of chrysanthemum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs). In a test of 6 AAs, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100 ng ml.−1 in chrysanthemum and tobacco expiants, respectively. Light conditions as well as expiant source and size also had significant effects. The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometric analyses revealed no endoduplication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli (8C or more), even at low AA concentrations (i.e., 5 to 10 μg mL-1).

The mechanism by which aminoglycoside antibiotics cause vasodilation of canine cerebral arteries.

The effects of aminoglycoside antibiotics were examined in canine cerebral arteries and in cultured cerebrovascular smooth muscle cells stimulated with oxyhemoglobin (OxyHb), a blood constituent which has been implicated in the pathogenesis of cerebrovascular spasm. In cerebral arterial rings precontracted with OxyHb (10 microM), the aminoglycosides caused a concentration-dependent decrease in isometric tension. The EC50s for the relaxation were 0.46+/-0.1 mM (n=6), 0.53+/-0.08 mM (n=12), 1.6+/-0.3 mM (n=7) and 3.9+/-0.5 mM (n=5) for neomycin, gentamicin, streptomycin and kanamycin, respectively. This order of potency corresponds approximately to the number of positive charges in the molecules. The aminoglycosides also inhibited the contractions to prostaglandin F2alpha (1 microM) and depolarizing concentrations of potassium chloride (60 mM). The order of potency was neomycin > gentamicin > streptomycin > kanamycin. The relaxation was maintained in vascular preparations denuded of endothelium. Neomycin (5 mM) abolished the Ca2+-independent contraction to PGF2alpha. In Fura-2-loaded cerebrovascular smooth muscle cells, OxyHb (1 microM) significantly enhanced the concentration of intracellular calcium ([Ca2+]i) by 330%. The administration of neomycin, gentamicin, kanamycin and streptomycin in concentrations corresponding to the EC50 from contractility studies, reduced the effects of OxyHb on [Ca2+]i by about 50% to 221+/-35 nM (n=7), 270+/-31 nM (n=7), 229+/-33 nM (n = 6) and 240+/-6 nM (n = 5), respectively. These results suggests that the effects of the aminoglycosides on the OxyHb-induced contraction and the long-term increase in [Ca2+]i, may arise from several effects, including inhibition of PLC, protection of calcium extrusion mechanisms, and interference with the process of [Ca2+]i accumulation.

Natural release of virulence factors in membrane vesicles by Pseudomonas aeruginosa and the effect of aminoglycoside antibiotics on their release

Pseudomonas aeruginosa (and various other gram-negative pathogens) liberate membrane vesicles during normal growth. These bilayered vesicles consist of endotoxin (lipopolysaccharide), outer membrane proteins and several potent hydrolytic enzymes including protease, alkaline phosphatase, phospholipase C and peptidoglycan hydrolase. The vesicles contain pro-elastase and alkaline phosphatase (which are periplasmic constituents) and so are important for packaging periplasmic components as they are liberated to the outside of the cell. Once liberated, the vesicles are capable of fusing with the membranes of epithelial cells and liberating their virulence factors into host cells where they degrade cellular components, thereby aiding infection by the pathogen. The aminoglycoside antibiotic, gentamicin, is thought to kill bacteria by inhibiting protein synthesis, yet this cationic antibiotic can also perturb the packing order of lipids, thereby destabilizing bilayered membranes. For pathogens with highly anionic lipopolysaccharide on their surface, such as P. aeruginosa, this membrane destabilization can be so serious that it can cause cell lysis; these cells are therefore killed by a combination of protein synthesis inhibition and surface perturbation. By destabilizing the membranes of P. aeruginosa, gentamicin increases the release of membrane vesicles three- to five-fold. This may help account for some of the bacterium-mediated toxicity encountered during patient treatment with aminoglycoside antibiotics.

Toxicokinetic approach for evaluating respiratory depression effect of aminoglycoside antibiotics: species differences in drug susceptibility.

The respiratory depression effect of aminoglycoside antibiotics was studied by a toxicokinetic approach, and species differences in drug susceptibility were elucidated based on plasma concentrations. An allometric relationship was obtained between total body clearance of arbekacin, a novel aminoglycoside antibiotic, and animal body weight. The power was 0.714, less than unity, which means smaller animals have higher ability to eliminate the drug from the body and need higher doses to attain a certain steady-state plasma concentration. When the infusion rate of arbekacin was altered, the total dose required to cause the toxicologic endpoint for respiratory depression (60 per cent loss of respiratory rates) changed greatly, but the plasma concentration of arbekacin at the toxicologic endpoint remained at almost a constant level. The concentration at the toxicologic endpoint was similar for all of the animal species examined and was 650-950 micrograms ml-1, even though the total dose required to cause the toxicologic endpoint varied greatly among the animal species. These findings suggest that the toxicologic effect compartment for respiratory depression is indistinguishable from the plasma compartment, and that species differences in the total dose are due to differences in pharmacokinetics of the drug, mainly in the total body clearance, but not to differences in intrinsic susceptibility to the drug.

Aminoglycoside antibiotics inhibit maxi-K + channel in single isolated cochlear efferent nerve terminals

Patch clamp recordings were obtained from isolated cochlear efferent nerve terminals. The effect of aminoglycoside antibiotics on single maxi-K + channels was determined. At positive voltages (cytosol with respect to extracellular side), neomycin, streptomycin, and kanamycin significantly reduced the single channel current amplitude of the maxi-K + channel from the cytosolic side. The IC 50 for neomycin was 9·10 −4 M from the cytosolic side and ⪢ 10 −3 M from the extracellular side. Streptomycin and kanamycin were less potent. No significant difference in inhibition of the single channel current amplitude by 2.5–10 −4 M cytosolic neomycin was observed between 7·10 −4 M and 10 −6 M free cytosolic Ca 2+. Neomycin had no significant effect on the open probability of the maxi-K + channel either from the cytosolic or from the extracellular side. These findings demonstrate that the maxi-K + channel in cochlear efferent nerve terminals can be a site of action for aminoglycoside antibiotics.

Effects of aminoglycoside antibiotics on the contractile response of guinea-pig vas deferens

Effects of aminoglycoside antibiotics on the contractile response of guinea-pig vas deferens

Effects of aminoglycoside antibiotics on the auditory brainstem response and post rotatory nystagmus in rats

Effects of three aminoglycoside antibiotics, amikacin (AMK), tobramycin (TOB), and gentamicin (GM), on the auditory and vestibular functions were assessed in rats, the most frequently used species in toxicity studies. Chronic electrodes for auditory brainstem response (ABR) recording were implanted on the epidural surface, and those for post rotatory nystagmus (PRN) were implanted at the nictitating membrane and the outer canthus. AMK, TOB, and GM were given intramuscularly twice daily for 3–4 weeks at a daily dose of 350, 150, and 100 mg/kg, respectively. The amplitude of each wave of the ABR was decreased or disappeared in the groups treated with AMK, TOB, and GM. In the PRN, the duration of the nystagmus was decreased in the TOB group and completely lost in the GM group. No abnormality was observed in the PRN in the AMK group. These results were similar to those reported in the ototoxicity studies of these drugs in guinea pigs and indicate that ototoxicity can be evaluated in rats as successfully as in guinea pigs by this procedure.

P-524 - Effects of aminoglycoside antibiotics on the thrombin-induced aggregation and intracellular Ca2+ mobilization of human platelets

P-524 - Effects of aminoglycoside antibiotics on the thrombin-induced aggregation and intracellular Ca2+ mobilization of human platelets