The effect of the earthworm Lumbricus rubellus on the bioavailability of cadmium and lead to the springtail Folsomia candida in metal-polluted field soils.

The bioavailability of metals can be influenced not only by soil properties but also by other species living at polluted sites. However, in laboratory experiments, usually only one test species is used to estimate bioavailability. In this study, a two-species approach was applied to assess the impact of the earthworm Lumbricus rubellus on the bioavailability of cadmium and lead to the springtail Folsomia candida using natural soils from a gradient of metal pollution. Earthworms were kept in half of the soil replicates for 4 weeks. Subsequently, the uptake and elimination kinetics of cadmium and lead in F. candida exposed for 21 days to the soils was determined. Earthworm activity affected soil properties but did not significantly affect metal uptake rate constants in springtails. The slightly higher uptake due to the presence of earthworms, which was consistent in all tested soils and for both metals, suggests that further research is needed on the role of species interactions in affecting metal bioavailability in soil.

AU-1 from Agavaceae plants causes transient increase in p21/Cip1 expression in renal adenocarcinoma ACHN cells in an miR-34-dependent manner.

Here, we show that AU-1, spirostanol saponin isolated from Agavaceae plants, causes a transient increase in cyclin-dependent kinase inhibitor (CDKI) p21/Cip1 through the upregulation of miRNAs, miR-34 and miR-21. AU-1 stimulated p21/Cip1 expression without exerting cytotoxicity against different types of carcinoma cell lines. In renal adenocarcinoma ACHN cells, AU-1 transiently elevated the expression level of p21/Cip1 protein without marked increases in p21/Cip1 mRNA levels. Rapid and transient increases in miR-34 and miR-21, both of which are known to upregulate p21/Cip1, were observed in AU-1-treated cells. Inhibitor for miR-34 and for miR-21 significantly blocked the AU-1-caused increase in p21/Cip1, indicating that elevation of p21/Cip1 protein by AU-1 is dependent on these microRNAs. We further clarified that NAD-dependent deacetylase SIRT1, a direct target of miR-34, is decreased by the treatment with AU-1. Furthermore, we found that SIRT1-knockdown increases p21/Cip1 protein levels in an miR-21-dependent manner. On the other hand, ectopic expression of p21/Cip1 resulted in the lowered expression of miR-34 and miR-21, suggesting that reciprocal regulation exists between p21/Cip1 and these miRNAs. We propose that the following feedback network composed of miR-34/SIRT1/miR-21/p21 is triggered by the treatment with AU-1: in cells treated with AU-1, transient elevation of miR-34 leads to the downregulation of SIRT1, thereby miR-21 is freed from SIRT1-dependent suppression. Then, elevated miR-21 upregulates p21/Cip1 protein, followed by the suppression of miR-34 expression.

[Discovery of Novel Biologically Active Compounds of Natural Origin, with a Focus on Anti-tumor Activity].

Numerous clinically valuable medicines, including anticancer drugs, have been developed from biologically active natural compounds and their structurally related derivatives. This review discusses novel natural compounds with promising biological activities and those with novel chemical structures. Glaziovianin A, an isoflavone isolated from the leaves of Ateleia glazioviana (Legminosae), inhibited cell cycle progression at the M-phase with an abnormal spindle structure. AU-1 and YG-1, 5ß-steroidal glycosides isolated from the whole plants of Agave utahensis and the underground parts of Yucca glauca (Agavaceae), induced apoptosis of HL-60 cells via caspase-3 activation. Lycolicidinol, an alkaloid isolated from the bulbs of Lycoris albiflora (Amaryllidaceae), induced transient autophagy and morphological changes in mitochondria in the early stage of the apoptotic cell death process in HSC-2 cells. Taccasterosides isolated from the rhizomes of Tacca chantrieri (Taccaceae) and stryphnosides isolated from the pericarps of Stryphnodendron fissuratum (Legminosae) are steroidal and triterpene glycosides with unique chemical structures having novel sugar sequences.

Application of capillary electrophoresis to the simultaneous determination and stability study of four extensively used penicillin derivatives

The applicability of capillary electrophoresis for the analysis of four extensively used penicillin derivatives (benzylpenicillin, ampicillin, amoxicillin, oxacilllin) has been studied. Because of structural similarities, the electrophoretic behavior of these derivatives is very similar; consequently an efficient separation using the conventional capillary zone electrophoresis is hard to be achieved. Their simultaneous separation was solved by using micellar electrokinetic capillary chromatography, the separation being based on the differential partition of the analytes between the micellar and aqueous phase. Using a buffer solution containing 25 mM sodium tetraborate and 100 mM sodium dodecyl sulfate as surfactant, at a pH of 9.3, applying a voltage of + 25 kV at a temperature of 25 °C, we achieved the simultaneous separation of the studied penicillin derivatives in less then 5 minutes. The separation conditions were optimized and the analytical performance of the method was evaluated in terms of precision, linearity, limit of detection, and quantification. Also, a simple capillary zone electrophoresis method was applied to study the stability of the studied penicillin derivatives in water at different temperatures, using ciprofloxacin hydrochloride as internal standard. It was observed that the extent of the hydrolysis of penicillins in water is highly dependent on the time and also temperature.

Estudou-se a aplicabilidade de electroforese capilar para a análise de quatro derivados de penicilina (benzilpenicilina, ampicilina, amoxicilina, oxacilina) amplamente utilizados. Em razão das semelhanças estruturais, o comportamento electroforético destes derivados é muito semelhante e, por conseguinte, a separação eficaz utilizando a electroforese capilar de zona convencional é difícil de ser efetuada. A separação simultânea foi realizada por cromatografia capilar electrocinética micelar, que se baseia na partição diferencial entre os analitos na fase micelar e aquosa. Utilizando-se solução tampão contendo 25 mM de tetraborato de sódio e 100 mM de dodecil sulfato de sódio, como agente tensioativo, com pH de 9,3, voltagem de +25 kV, à temperatura de 25 °C, obteve-se a separação simultânea das penicilinas estudadas em menos de 5 minutos. As condições de separação foram otimizadas e o desempenho do método analítico foi avaliado em termos de precisão, linearidade, limite de detecção e de quantificação. Além disso, aplicou-se método de electroforese capilar de zona simples para estudar a estabilidade de penicilinas em água a diferentes temperaturas, utilizando cloridrato de ciprofloxacino como padrão interno. Estabeleceu-se que o grau de hidrólise de penicilinas em água é altamente dependente do tempo e também da temperatura.

Ferrocenyl bioconjugates of ampicillin and 6-aminopenicillinic acid--synthesis, electrochemistry and biological activity.

We report on the synthesis of ferrocenyl-ampicillin and ferrocenyl-6-aminopenicillinic acid bioconjugates. Title compounds were characterized by (1)H NMR, IR, MS and elemental analysis. These novel ferrocenyl-antibiotic conjugates were also investigated by cyclic voltammetry (CV). Ferrocenyl-ampicillin complexes revealed reversible uncomplicated oxidation whereas ferrocenyl-6-aminopenicillinic acid derivatives were found to exhibit adsorption waves in cathodic scans. Antibacterial activities of our ferrocenyl-antibiotic conjugates against Gram-positive methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), vancomycin-resistant S. aureus (VRSA) and Staphylococcus epidermidis bacterial strains were determined. Our experiments show significant antibacterial activity of ferrocenyl-6-aminopenicillinic acid bioconjugates against the bacterial strains tested. Contrary to that ferrocenyl-ampicillin derivatives were inactive. The inhibitory effects on the dd-carboxypeptidase 64-575 II exerted by our ferrocenyl-6-aminopenicillinic acid bioconjugates were established in the low nanomolar range. The tumor cell growth inhibition of representative ferrocenyl-ampicillin and ferrocenyl-6-aminopenicillinic acid bioconjugates against MCF-7 breast adenocarcinoma and HT-29 colon carcinoma cell lines were studied in vitro. Similar to the antibacterial activity tests the assays in tumor cells revealed significant antiproliferative effects exerted by ferrocenyl-6-aminopenicillinic acid bioconjugates.

Evaluation of epitope tags for protein detection after in vivo CNS gene transfer.

Functional characterization of disease-related proteins, their splice variants and dominant negative mutants in the context of complex CNS tissues such as brain and retina is frequently assessed by in vivo gene transfer. For correct interpretation of results it is imperative that the protein under investigation is unambiguously detected in the transduced cell types and can be distinguished from any endogenously expressed physiological variants. Therefore the first systematic evaluation of epitope tags used to trace ectopically expressed proteins in the central nervous system is presented here. Substantial differences in the performances of various epitope tag-antibody combinations with respect to sensitivity, specificity and influence of the epitope tag on the fusion protein are elucidated. Epitope tags already established for protein detection in vitro and to some extent in vivo (c-Myc, HA and FLAG tags) were immunohistochemically detected with high sensitivity. However, detection of these tags revealed problems with background staining and we also document structural and functional influence of the tags on the fusion protein. In order to prevent such unwanted side-effects, epitope tags which have not yet been used for in vivo applications (IRS, EE and AU1 tags) were characterized in brain, retina and cultured neurons. While use of the IRS and EE tags was hindered by low sensitivity or specificity, optimal results were obtained with the AU1 epitope, which may develop into a standard tool for detection of ectopic protein expression in the central nervous system.

Localized bacillary angiomatosis in the oral cavity: observations about a neoplasm with atypical behavior. Description of a case and review of the literature.

Bacillary angiomatosis is a rather frequent infectious pathology appearing mainly in the skin but can also affect the liver, spleen, heart, bones, lungs, muscles, central nervous system and other organs. The localization of the lesion in the oral cavity is rather rare, as it is evident in the literature. Bacillary angiomatosis can be clinically similar to the Kaposi's sarcoma and histologically confused with angiosarcoma, epitheloid hemangioma and pyogenic granuloma. A case of bacillary angiomatosis of the oral cavity in an immuno-competent patient is described. The high tendency to relapse, the capability in migration and to involve several localizations at the same time have induced the authors to deepen the research to exclude the possibility that it could be a Kaposi's sarcoma or a pyogenic granuloma and to get to an accurate diagnosis in order to resolve the disease.

[Oesophageal ulcer associated with the use of bacampicillin]. / Ulcère oesophagien secondaire à la prise de bacampicilline.

INTRODUCTION: Many drugs may provoke oesophageal disorders. In France, doxycyclin, potassium chloride non-steroidal anti-inflammatories are the most frequent agents incriminated. OBSERVATION: A 27 year-old man consulted for dysphagia and odynophagia due to oesophagitis following administration of bacampicillin. COMMENTS: Several drugs can provoke oesophageal disorders including ulcerations. Predisposing circumstances exist. Ingestion of the drugs with a sufficient quantity of water (60 to 100 ml), whilst standing or sitting should be able to limit this type of event.

Intracellular accumulation and activity of ampicillin used as free drug and as its phthalimidomethyl or pivaloyloxymethyl ester (pivampicillin) against Listeria monocytogenes in J774 macrophages.

AIMS: To determine the intracellular accumulation in a macrophage cell line of ampicillin and ampicillin esters, and to measure their activity against intracellular Listeria monocytogenes. METHODS: Quantitative evaluation of the activity of ampicillin, phthalimidomethylampicillin (PIMA) or pivaloyloxymethylampicillin (PIVA) against intracellular L. monocytogenes, and direct measurement of cellular ampicillin concentration in J774 macrophages. RESULTS: Ampicillin, PIMA and PIVA caused a 0.5 log decrease in cell-associated cfu within 5 h when used at an extracellular concentration of 3.6 microM [10 x MIC of ampicillin (1.25 mg/L); 1.83 mg/L for PIMA and 1.67 mg/L for PIVA]. Addition of beta-lactamase in the extracellular milieu abolished the activity of ampicillin and of PIMA but not that of PIVA. At low extracellular concentrations [0.5 x MIC ampicillin (62.5 microg/L); equimolar concentrations for PIMA (91.5 microg/L) and PIVA (83.5 microg/L)], ampicillin and PIMA lost all activity (compared with controls), but PIVA remained as active as at the higher concentration. Incubation of cells with PIVA at the low concentration (83.5 microg/L) for 20 h caused a 2 log reduction of cfu if the medium was changed every 5 h (to compensate for the degradation of extracellular PIVA). Incubation of cells with PIVA allowed for a marked (four- to 25-fold) cell accumulation of ampicillin, whereas no ampicillin accumulation was seen for cells incubated with ampicillin or with PIMA. CONCLUSIONS: This is the first demonstration that PIVA (a prodrug of ampicillin) can be used to promote ampicillin cellular accumulation and, thereby to increase ampicillin intracellular activity. PIVA could be useful for control of the intracellular multiplication of L. monocytogenes.

Cell handling, membrane-binding properties, and membrane-penetration modeling approaches of pivampicillin and phthalimidomethylampicillin, two basic esters of ampicillin, in comparison with chloroquine and azithromycin.

PURPOSE: The purpose of this work was to examine and understand the cellular pharmacokinetics of two basic esters of ampicillin, pivaloyloxymethyl (PIVA) and phthalimidomethyl (PIMA), in comparison with lysosomotropic drugs (chloroquine, azithromycin). METHODS: Cell culture studies (J774 macrophages) were undertaken to study uptake and release kinetics and to assess the influence of concentration, pH, proton ionophore (monensin), and MRP and P-gp inhibitors (probenecid, gemfibrozil, cyclosporin A, GF 120918). Equilibrium dialysis with liposomes were performed to directly asses the extent of drug binding to bilayers. Conformational analysis modeling of the drug penetration in bilayers was conducted to rationalize the experimental observations. RESULTS: PIVA and PIMA showed properties in almost complete contrast with those of chloroquine and azithromycin, i.e., fast apparent accumulation and fast release at 4 degrees C as well as at 37 degrees C, saturation of uptake (apparent Kd 40 microM), no influence of monensin, MRP, or P-gp inhibitors; tight binding to liposomes (Kd approx. 40 microM); and sharp increase in calculated free energy when forced in the hydrophobic domain. CONCLUSIONS: Although they are weak organic bases, PIVA and PIMA show none of the properties of lysosomotropic agents. We hypothesize that they remain locked onto the pericellular membrane and may never penetrate cells as such in significant amounts.

Monitoring of ampicillin and its related substances by NMR.

A 1H NMR procedure for the monitoring of ampicillin (Amp) and its main related substance in different media, has been developed. The characteristics peak of Amp, 6-aminopenicillanic acid (6-APA), phenylglycine (PhG), and penicilloic acid in the range of 0.5-0.9 and 3.0-4.5 ppm were used for their identification in drugs and serum samples. The quantitative works were performed based on the intensity of protons of the methyl group link to the beta-lactam cyclic of Amp and 6-APA and the aromatic protons of PhG relative to the protons of methylene group of maleic acid, as internal standard, at constant temperature. The resulting data are compared with those obtained with an HPLC method proposed by British Pharmacopoeia. Statistical studies show that, at a confidence limit of %95, there is no significant difference between the two methods. In comparison with the HPLC method, the proposed NMR method does not require any sample pretreatment, standard solution preparation, long analysis time and use of any carcinogenic solvent. The method was applied to the determination of Amp and its related substances in synthetic mixtures, drug powders and serum samples.

Differences in postoperative morbidity rates, including infection and dry socket, and differences in the healing process after mandibular third molar surgery in patients receiving 1-day or 3-day prophylaxis with lenampicillin.

The aim of this study was to examine the effect of antibiotic prophylaxis for mandibular third molar surgery from the viewpoint of the duration of administration. A comparative study was conducted on postoperative infection, dry socket, and the healing process in 178 healthy patients receiving 1-day or 3-day prophylaxis with lenampicillin (LAPC; 1.5 g/day). Postoperative infection developed in only 1 (1.1%) of the 91 patients in the 3-day group, while there was no patient with infection in the 87 patients in the 1-day group. Dry socket developed in 8.0% of the 87 patients in the 1-day group and in 7.7% of the 91 patients in the 3-day group. However, the incidence of these complications, in relation to the degree of impaction of the molars, did not significantly differ between the 1-day and 3-day groups. Scores indicating clinical symptoms for the 7 days after surgery also reflected no significant difference between the groups, irrespective of the degree of impaction. These results suggest that 1-day therapy with LAPC in our regimen may at least be recommended as a prophylaxis for mandibular third molar surgery in medically healthy patients.

Incidence of deep fascial space infection after surgical removal of the mandibular third molars.

Nine hundred and ninety-three patients who underwent surgical removal of the mandibular third molars with oral antibiotic prophylaxis were examined to determine the incidence of postoperative deep fascial space infection and its background factors. Postoperative deep fascial space infection was observed in 8 of the patients (0.8%; 4 males and 4 females), and submandibular spaces were involved in all infected patients. Only 1 of these 8 patients was an immune compromised host. Patients aged 30 years or more had a significantly higher incidence of deep fascial space infection than those aged under 30. Five patients had partial bony impactions and 3 had complete bony impactions. However, the incidence of infection according to the molar positions was not significantly different between partial bony impaction and complete bony impaction. The 8 patients had not had pericoronitis preoperatively. The clinical courses of all were favorable after antibiotics were administered intravenously. In conclusion, the incidence of deep fascial space infection after removal of the mandibular third molars was low, at 0.8%. However, it may be desirable to remove the molars, if applicable, at a younger age because of the higher incidence of infection in patients aged over 30. The results of this study also offer information that will be useful as a basis for obtaining informed consent from patients whose mandibular third molars are to be removed.

Pustular drug eruption due to bacampicilin hydrochloride in a patient with psoriasis.

We report a case of pustular drug eruption due to bacampicilin hydrochloride which developed in a patient with pustular psoriasis. The patient was a 45-year-old Japanese woman with psoriasis which started as pustular psoriasis twenty years previously. In 1994, she developed generalized erythema with pustules accompanied by high fever and liver injury. Clinical and histological findings of this pustular eruption were different from her previous episodes of pustular psoriasis. Erythemas and pustules disappeared and her abnormal transaminase returned to normal rapidly when she discontinued bacampicilin hydrochloride. Her positive reaction to a patch test and a lymphocyte stimulation test also suggested that our case had a pustular drug eruption rather than pustular psoriasis induced by a drug.

Ampicillin concentrations in radicular cysts following a single oral administration of bacampicillin.

1. Ampicillin concentrations in cyst wall (wall) and cyst fluid (fluid) of radicular cyst and serum following a single oral administration of bacampicillin (equivalent to 500 mg of ampicillin) were measured by a paper disk method. 2. The mean peak concentrations of ampicillin in wall, fluid, and serum occurred at identical times, 1.5 hr, and were 2.39 micrograms/g, 0.77, and 10.24 micrograms/ml, respectively. 3. Mean ampicillin concentration ratios of wall/serum, fluid/serum, and fluid/wall at the peak time were 0.23, 0.07, and 0.40, respectively. 4. Mean ampicillin concentrations in wall and fluid at the peak time exceeded MIC (minimum inhibitory concentration) for 90% (0.5 microgram/ml) for clinically isolated strains of alpha-hemolytic Streptococci.

[Prophylactic use of antibiotics for fever following fiberoptic bronchoscopy and bronchography].

To evaluate the effect of bacampicillin hydrochloride on fever following fiberoptic bronchoscopy and bronchography, we conducted multi-institutional randomized study. In bronchographic examinations, the rise of body temperatures in the bacampicillin group (0.82 +/- 0.13 degrees C: mean +/- SE) was significantly smaller than that in the control group (1.39 +/- 0.25 degrees C) on the second day of examination. Bacterial infection may contribute to the rise of temperature on the day following bronchography, but no pneumonia or sepsis was observed. There was no differences in the rise of body temperature on the first, third or fourth day. In fiberoptic bronchoscopic examinations, there was no difference between the two groups. We conclude that there is no clinical indication of the value of prophylactic use of antibiotics in either fiberoptic bronchoscopy or bronchography.

Ampicillin concentrations in human radicular granuloma following a single oral dose of bacampicillin.

Ampicillin concentrations in human serum and radicular granulomas of 42 patients were determined after a single oral dose of bacampicillin (equivalent to 500 mg of ampicillin). Although wide variations were found among both serum and radicular granuloma ampicillin concentrations, measurable concentrations were found in all cases. The mean peak ampicillin concentrations in serum and radicular granulomas occurred at identical times, 1.5 hours, and were 11.19 micrograms/mL (range, 1.30 to 21.00 micrograms/mL) and 5.12 micrograms/g (range, 0.50 to 10.50 micrograms/g), respectively. The mean radicular granuloma/serum ampicillin concentration ratio at the peak time was 0.42. Ampicillin concentrations in radicular granulomas exceeded most of the minimum inhibitory concentrations for bacteria commonly isolated from odontogenic infections.

Ampicillin concentrations in human oral tissues following a single oral administration of lenampicillin.

1. Ampicillin concentrations in serum (n = 20), gingiva (n = 12), jawbone (n = 13), dental follicle (n = 12), radicular granuloma (n = 2) and radicular cyst (n = 2) were measured in specimens obtained during 0.5-2.5 hr after a single oral administration of lenampicillin (equivalent to 500 mg of ampicillin). 2. Measurable ampicillin concentrations were found in all serum and tissues. 3. Ampicillin concentrations in serum and tissues except for some gingiva and jawbone exceeded MIC for 90% of clinically isolated strains of alpha-hemolytic Streptococci. 4. Ampicillin concentrations in gingiva and jawbone were below the MIC for 90% in 2 out of 12 and 4 out of 13 specimens, respectively.

Effect of bacampicillin and ampicillin on the phagocytic, microbicidal functions of human macrophages.

During acute bacterial infections, antibiotics only assist the phagocytic system in eradicating the invading organism. In this regard, the effect of ampicillin and its prodrug bacampicillin on the phagocytosis and killing of Staphylococcus aureus by human macrophages was compared. Bacterial phagocytosis was enhanced after pre-incubation of staphylococci with bacampicillin. At a concentration of half the MIC bacampicillin showed a much more impressive killing capacity on macrophage-ingested live bacteria than did ampicillin, under all the experimental conditions tested.