MSC-derived exosomes protect auditory hair cells from neomycin-induced damage via autophagy regulation

BACKGROUND: Sensorineural hearing loss (SNHL) poses a major threat to both physical and mental health; however, there is still a lack of effective drugs to treat the disease. Recently, novel biological therapies, such as mesenchymal stem cells (MSCs) and their products, namely, exosomes, are showing promising therapeutic potential due to their low immunogenicity, few ethical concerns, and easy accessibility. Nevertheless, the precise mechanisms underlying the therapeutic effects of MSC-derived exosomes remain unclear. RESULTS: Exosomes derived from MSCs reduced hearing and hair cell loss caused by neomycin-induced damage in models in vivo and in vitro. In addition, MSC-derived exosomes modulated autophagy in hair cells to exert a protective effect. Mechanistically, exogenously administered exosomes were internalized by hair cells and subsequently upregulated endocytic gene expression and endosome formation, ultimately leading to autophagy activation. This increased autophagic activity promoted cell survival, decreased the mitochondrial oxidative stress level and the apoptosis rate in hair cells, and ameliorated neomycin-induced ototoxicity. CONCLUSIONS: In summary, our findings reveal the otoprotective capacity of exogenous exosome-mediated autophagy activation in hair cells in an endocytosis-dependent manner, suggesting possibilities for deafness treatment.

Neomycin: sources of contact and sensitization evaluation in 1162 patients treated at a tertiary service

Abstract Background: Neomycin is used in several over-the-counter pharmaceutical formulations in Brazil. In Europe and Canada, where it is not freely available, its sensitization frequency is lower than in the United States, where this does not occur. Objective: To present the frequency of sensitization to neomycin observed in a tertiary hospital and the pharmaceutical formulations sold in Brazil containing neomycin. Method: Retrospective analysis of positive results to neomycin, obtained through patch tests performed in a tertiary hospital, from 2009 to 2018 and investigation of topical drugs and vaccines containing neomycin in Brazilian databases available on the internet. Results: Among 1,162 patients, 71 (6%) had positive reactions to neomycin, 65% female and 35% male individuals, 46% were over 50 years old, and 24% had a personal history of atopy. The dermatitis lasted from four months to 20 years. Lesions were located in 69% of the patients on the upper limbs, in 55% they were on the lower limbs, and in 42% they were disseminated in more than 4 sites. Polysensitization was detected in 55% of cases. Of these, 28% were linked to sensitization to rubber allergens and 27% to potassium bichromate. A total of 158 topical presentations of neomycin were found: 79 ointments, 58 creams, 10 ophthalmic solutions, seven otological solutions, one oral solution, two nasal solutions, and one antiseptic powder, in addition to 11 types of vaccines. Study limitations: Retrospective study. Conclusion: Sensitization to neomycin occurred in 6% of the studied population, affecting more females aged over 50 years, with skin lesions located mainly on the upper and lower limbs, in the context of chronic contact dermatitis. Neomycin was found in 135 formulations, most of them available over the counter, as well as in 11 miscellaneous vaccines.

Study on the Quality Control Standard of Lvxintong Rugao / 药学实践与服务

Objective To establish a quality control method for Lvxintong Rugao. Methods Ketoconazole, Halcinonide and Neomycin sulfate were identified by TLC. The content of Ketoconazole and Halcinonide were determined by HPLC. The chromatographic column of Agilent ZORBAX SB-C18 (4.6 mm×150 mm, 5 μm) column was used. Methanol-phosphate buffer (pH=7.40, 75:25) was applied as the mobile phase. The detection wavelength was 235 nm. The flow rate was 1.0 ml/min and the column temperature was set at room temperature. Neomycin Sulfate was determined by polarimetric analysis. Results The identification and determination methods showed good specificity. Ketoconazole and Halcinonide displayed good linearity within the range of 1.999~39.98 μg (r=0.999 9) and 0.400 8~8.016 μg (r=0.999 9), respectively. The average recoveries were 97.75% (RSD 0.77%) and 97.57% (RSD 0.84%), respectively. For the determination of Neomycin Sulfate, r=0.999 6 (n=6) in the range of 130.4~2 608 U/ml (n=6). The precision and repeatability of RSD were 1.1% and 1.6%, respectively. The solutions were stable in 6 h and the average recovery was 98.8% (RSD 2.6%). Conclusion The method could be used as the quality control method for Lvxintong Rugao.

Dermatite de contato agravada por outra sensibilidade em automedicação / Contact dermatitis aggravated by other sensitivity in self-medication

RESUMO: Dermatite alérgica de contato é uma doença cutânea inflamatória, não infecciosa, cuja base do tratamento é a identificação e eliminação do agente causal. Cocoamidopropil betaína é um surfactante muito utilizado nos produtos de uso pessoal, notadamente de uso capilar. Essa substância não está presente na bateria padrão brasileira. Neomicina é um antibiótico usado em preparações tópicas. Objetivamos mostrar paciente que desenvolveu alergia no couro cabeludo e que fez erroneamente automedicação com produto que continha substância a qual era ainda mais sensível. O caso é de uma mulher, 36 anos, evoluindo há dois meses com eczema pruriginoso, em áreas de implantação capilar e nuca. Relatava progressiva piora. Diante da suspeita de dermatite de contato, foi realizado teste de contato, utilizando-se da bateria padrão Latino-Americana. Com 96 horas (D4) evidenciou-se positividade leve (+) para cocoamidopropil betaína e forte (++) para neomicina. O resultado positivo para a neomicina foi intrigante, uma vez que a observação do rótulo dos produtos de uso pessoal não a continha. Diante do resultado do teste, após ser questionada novamente, ela confirmou a omissão da automedicação diária com pomada de neomicina. Em conclusão, mostramos a alergia a produtos de uso capilar. Reforçamos a necessidade de se fazer um teste de contato com bateria padrão atualizada. Por fim, alertamos sobre o risco da automedicação. (AU)

ABSTRACT: Allergic contact dermatitis is an inflammatory, non-infectious skin disease. The treatment is based on the identification and elimination of the causal agent. Cocamidopropyl betaine is a surfactant widely used in products for personal use, especially capillary use. This substance is not present in the Brazilian baseline series. Neomycin is an antibiotic used in topical preparations. We aimed to show a patient who developed na allergy in the scalp and mistakenly self-medicated with a product that contained a substance to which it was even more sensitive. The case is of a woman, 36 years old, evolving for 2 months with pruritic eczema, in areas of capillary and nape implantation. She reported progressive worsening. When contact dermatitis was suspected, a contact test was performed using the Latin American baseline series. At 96 hours (D4) there was mild positivity (+) for cocamidopropyl betaine and strong (++) for neomycin. The positive result for neomycin was intriguing, since the observation of the label of products for personal use did not contain it. In view of the test result, after being questioned again, she confirmed the omission of daily self-medication with neomycin ointment. In conclusion, we showed the allergy to hair products. We reinforced the need for an updated baseline series patch test. Finally, we warned about the risk of self-medication. (AU)

Development and validation method for simultaneous quantification of neomycin and polymyxin B by HPLC-ELSD and comparison with microbiological method

The aim of this study is to develop the first simultaneous method for quantification of neomycin and polymyxin B inthe presence of dexamethasone using High Performance Liquid Chromatography (HPLC) with an Evaporative LightScattering Detector (ELSD). The analysis was performed using a phenyl Waters X Bridge column, an evaporationtemperature of 50oC, and a nitrogen pressure of 320 kPa. The mobile phase consists of a combination of methanoland trichloroacetate acid (40 mM, pH 1.70–1.80) in gradient mode, flow rate at 1.0 ml/minute, detector gain of 6,and analysis time of 35 minutes. The linearity was achieved with a concentration of 100–500 µg/ml (r = 0.99955)for neomycin and concentration of 30–100 µg/ml (r = 0.99703) for polymyxin B. Recovery results were obtainedbetween 99.150% and 104.773% for neomycin and 96.538% and 105.139% for polymyxin B. The analysis samplefrom the market was found to be 102.27% for neomycin and 100.79% for polymyxin B. The result was compared tothe standard microbiological method. Based on the T-test results of two samples with a 95% confidence level (α =0.05), it was concluded that there was no significant difference between HPLC-ELSD and microbiological methodsfor determining neomycin and polymyxin B. The HPLC-ELSD method has a potential for routine analysis due toadvantages in terms of increasing precision, accuracy, and shorter testing time.

Transition of Determination Method of Potency of Neomycin Sulfate from Microbiological Assay to HPLC-PAD / 中国药学杂志

OBJECTIVE: To establish a high performance liquid chromatography combined with pulsed amperometric detection(HPLC-PAD)method for determination of potency of neomycin sulfate. METHODS: An improved HPLC-PAD method from EP method for determination of the content and related substances of neomycin sulfate was established and validated. The study of impurity profile of neomycin sulfate was completed by LC-IT-TOF method with the help of on-line desalination using a suppressor; and the main components in neomycin sulfate were clarified combining the RESULTS of impurity profile and minimum inhibitory concentrations of the main components and impurities. The semi-preparative liquid chromatography-evaporative light scattering detector(ELSD) was self-assembled, highly purified neomycin B and neomycin C were prepared and their structural confirmation was also conducted. The contents of highly purified neomycin B and neomycin C were determined by means of mass balance method. The potencies of highly purified neomycin B and neomycin C were determined by three-dose antibiotic microbial assay and the conversion factors between contents of neomycin B and neomycin C and their potencies were calculated separately and then a formula for the calculation of potency of neomycin sulfate from the content of main components of neomycin B and neomycin C was obtained.At last, a verification experiment for the accuracy of the conversion factor and the formula were designed and a serial of tests were carried out to investigate the interaction and the verification for the actual sample. RESULTS: The improved HPLC-PAD method was superior to the European Pharmacopoeia method in the separation ability and stability, and was suitable for accurate quantification of various components of neomycin sulfate and related substance inspection. The successful removal of trifluoroacetic acid in the mobile phase by the technology of desalination on-line using a suppressor broke a new way for the study of impurity profile of aminoglycoside such as neomycin sulfate. Combining the impurity profile with the RESULTS of MIC it was clarified that the main activity components of neomycin sulfate were neomycin B and neomycin C. Highly purified neomycin B and neomycin C were successfully prepared. A conversion factor for the transition from potency to purity of neomycin sulfate was obtained through experiments and calculations and was verified successfully. CONCLUSION: It is feasible to replace the microbial assay by HPLC-PAD method for determining the potency of neomycin sulfate.

Determination of the content and related substances of neomycin sulfate by HPLC combined with pulsed amperometric detection / 中国药科大学学报

@#A novel method was developed for the content assay and related substances determination of neomycin sulfate by high performance liquid chromatography combined with pulsed amperometric detection(HPLC-PAD). The HPLC was performed on Thermo AcclaimTMAmG C18(4. 6 mm×150 mm, 3 μm). The mobile phase consisted of aqueous solution with 2% trifluoroacetic acid containing 0. 01% pentafluoropropionic acid and 0. 6%NaOH. The pulsed amperometric detector was operated with aquadruple-potential waveform for the detection. Neomycin B, Neomycin C and thirteen related substances were adequately separated by the established HPLC conditions. The limits of detection(LOD)and quantification(LOQ)of neomycin B and neomycin C were both 1. 75 ng and 3. 5 ng, respectively. Good linearities of neomycin B and neomycin C were found in their respective ranges which their correlation coefficients were greater than 0. 998 5. The established method is characterized by high specificity, sensitivity and wide range of linearity which has a good application prospect and provides the basis for improving the standard and quality control of neomycin sulfate.

Desenvolvimento de método colorimétrico para determinação de sulfato de neomicina empregando os conceitos de qualidade por design analítico (AQbD) / Development of a colorimetric method for determination of neomycin sulphate using the concepts of quality by analytical design (AQbD)

Para efetivamente tratar uma infecção, é necessário que o antibiótico possua atividade antimicrobiana adequada e seja capaz de inibir o crescimento do microrganismo patogênico. O doseamento microbiológico é uma metodologia indicada para a análise do antimicrobiano de forma simples, quando comparado com outras metodologias. A Food and Drug Administration (FDA) tem encorajado uma abordagem proativa para introduzir inovações e benefícios associados ao processo de produção farmacêutica. A Qualidade por Design Analítico (AQbD) ajuda no desenvolvimento de métodos analíticos robustos e de baixo custo, que são aplicáveis durante todo ciclo de vida do produto. Os métodos microbiológicos tradicionais, de forma geral, apresentam baixa reprodutibilidade e alta incerteza. Desta forma, justifica-se o desenvolvimento de métodos microbiológicos alternativos para a análise de antimicrobianos empregando-se os conceitos de Qualidade por Design Analítico, com a finalidade de melhorar a reprodutibilidade e reduzir a incerteza final. O objetivo deste trabalho foi aplicar o conceito de Qualidade por Design Analítico (AQbD) no desenvolvimento de método colorimétrico para análise de sulfato de neomicina. O sulfato de neomicina é um antimicrobiano aminoglicosídeo amplamente empregado no tratamento de infecções cutâneas ou mucosas, tais como queimaduras, úlceras, e dermatites infecciosas. Métodos cromatográficos como a cromatografia líquida de alta eficiência em fase reversa, de pareamento iônico ou cromatografia iônica com derivatização (pré ou pós-coluna) são utilizados para a análise de aminoglicosídeos, inclusive sulfato de neomicina. Contudo, de acordo com as farmacopeias, o método microbiológico é o método analítico de escolha para a análise de sulfato de neomicina e outros aminoglicosídeos. A análise colorimétrica é um método amplamente utilizado para a detecção e quantificação de diferentes substâncias, incluindo o crescimento microbiano em estudos de eficácia terapêutica. Neste trabalho, propomos o uso de resazurina como marcador colorimétrico. O indicador sofre uma reação de oxido-redução na qual altera a coloração em resposta à redução química resultante do crescimento celular. O uso de microplacas para a análise colorimétrica é uma alternativa ao método realizado em tubos de ensaio. Uma alternativa ao uso de espectrofotômetros para a análise colorimétrica é o uso de aparelhos smartphones, pois são equipados com CPUs rápidas, câmeras de alta resolução e sensores de imagem. O processamento da imagem captada pela câmera do dispositivo é utilizado como um analisador colorimétrico. Portanto, a aplicação dos conceitos de Qualidade por Design Analítico (AQbD) possibilitou o desenvolvimento racional de método microbiológico colorimétrico para análise de sulfato de neomicina

o effectively treat an infection, the antibiotic must have adequate antimicrobial activity and be capable of inhibiting the growth of the pathogenic microorganism. The microbiological assay is an indicated methodology for the analysis of the antimicrobial in a simple way, when compared with other methodologies. The Food and Drug Administration (FDA) has encouraged a proactive approach to introducing innovations and benefits associated with the pharmaceutical production process. Analytical Design Quality (AQbD) assists in the development of robust, low cost analytical methods that are applicable throughout the product life cycle. Traditional microbiological methods, in general, have low reproducibility and high uncertainty. Thus, it is justified the development of alternative microbiological methods for the analysis of antimicrobials using the concepts of Quality by Analytical Design, in order to improve reproducibility and reduce final uncertainty. The objective of this work was to apply the concept of Quality by Analytical Design (AQbD) in the development of a colorimetric method for the analysis of neomycin sulfate. Neomycin Sulfate is an aminoglycoside antimicrobial widely used in the treatment of cutaneous or mucosal infections, such as burns, ulcers, and infectious dermatitis. Chromatographic methods such as reverse phase high performance liquid chromatography, ion-pairing or ion chromatography with derivatization (pre or post-column) are used for the analysis of aminoglycosides, including neomycin sulfate. However, according to pharmacopoeias, the microbiological method is the analytical method of choice for the analysis of neomycin sulphate and other aminoglycosides. Colorimetric analysis is a widely used method for the detection and quantification of different substances, including microbial growth in studies of therapeutic efficacy. In this work, we propose the use of resazurin as a colorimetric marker. The indicator undergoes an oxidation-reduction reaction in which it alters the coloration in response to the chemical reduction resulting from cell growth. The use of microplates for colorimetric analysis is an alternative to the method carried out in test tubes. An alternative to the use of spectrophotometers for colorimetric analysis is the use of smartphones because they are equipped with fast CPUs, high resolution cameras and image sensors. The image processing captured by the device's camera is used as a colorimetric analyzer. Therefore, the application of the concepts of Quality by Analytical Design (AQbD) allowed the rational development of a microbiological colorimetric method for analysis of neomycin sulfate

Scoring Of Central Neurotoxicities of Aminoglycoside Antibiotics – An Experimental Study in Albino Rats.

Background: Aminoglycoside antibiotics are still the drug of choice in variable conditions like resistant tuberculosis and septicaemia. Toxic effects are the greatest hurdle in their liberal use. Their central neuro-toxicities specially in terms of affinity are yet to be explored. Methods: Experimental rats received streptomycin, kanamycin and gentamycin in a dose of 30mg/kg, 400mg/kg and 135 mg/kg respectively, IMI, daily for 21 days. Total lipids, phospholipids, cholesterol and gangliosides were estimated in auditory cortex, medial geniculate body, inferior colliculus, cerebellum and spinal cord in both control and experimental rats. Results: On the basis of statistically significant alterations in aforementioned biochemical parameters, affinity of drugs was quantified by scoring. Streptomycin and kanamycin showed maximum toxicity in terms of scoring of 4 with preferential targets i.e. medial geniculate body and inferior colliculus respectively. Gentamycin showed affinity for higher centres only with equal scoring of 3 for toxicity at three locations i.e. auditory cortex, medial geniculate body and inferior colliculus. Conclusion: Such preferential toxicities might reflect some aspects of mechanism of toxicity of different drugs.

Analysis of antibiotic residues in milk from healthy dairy cows treated with bovine mastitis ointment using ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry

This study was conducted to analyze penicillin G (PEG), streptomycin (STR) and neomycin (NEO) residues in milk of healthy lactating cows. Milk samples were collected from all four quarters of 12 dairy cows 2–7 days after intramammary infusions of an ointment containing PEG, STR and NEO once (n = 4; group I) or twice (n = 4, group II) daily. Ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry was used to determine the antibiotic residues in the samples. The correlation coefficient (r 2) of the calibration curves for all antibiotics was > 0.999 and the limits of detection and quantification were 0.002–0.005 µg/mL and 0.007–0.02 µg/mL, respectively. Recovery rates were ranged from 75.5 to 92.3%. In group I, PEG, STR and NEO residues were detected in milk at 2, 3 and 2 days post-treatment, respectively, which were below the maximum residue limit (MRL). In group II, PEG, STR and NEO residues were detected in milk at 2, 3 and 3 days post-treatment, respectively, which were bellow the MRL. These results suggest that a 3-day for milk withdrawal period after the ointment treatment might be sufficient for reduction of the antibiotic residues below the MRL.

Effect of covalent attachment of neomycin on conformational and aggregation properties of catalase.

The carboxylic groups of glutamic acid and aspartic acid residues of catalase (CAT) were chemically modified using the treatment of the enzyme with 1-ethyl-3-(3'-dimethylamino) carbodiimide hydrochloride (EDC) and neomycin. The effect of covalent attachment of neomycin on the enzymatic activity, conformational and aggregation properties of CAT was investigated. The modification of CAT with different concentrations of neomycin showed two different types of behavior, depending up on the concentration range of neomycin. In the concentration range from 0.0 to 5.2 mM, neomycin-modified CAT, compared to the native enzyme exhibited higher α-helix content, reduced surface hydrophobicity, little enhancement in CAT activity and a better protection against thermal aggregation, whereas at concentrations greater than 5.2 mM, the modified enzyme exhibited a significant decrease in CAT activity and an increase in random coil content which may result in disorder in the protein structure and increase in thermal aggregation. This modification is a rapid and simple approach to investigate the role of aspartate and glutamate residues in the structure, function and folding of CAT.

Weakening resistance marker for establishing a process of screening high-producing CHO cell lines / 中国药科大学学报

To optimize Chinese hamster ovary (CHO)expression system and establish a process of screening CHO cell lines with high productivity;neomycin-phosphotransferase (NPT)expressed by the resistance marker gene on the expression vector was mutated with amino acid D at 261 changed to G.After selection by culturing with G418;the survival rate of CHO cells bearing mutant-NPT was significantly lower than that of the cells bear-ing wide type NPT.An enhanced green fluorescent protein (EGFP)was genetically linked to the N terminus of the IgG1 Fc fragment part to generate an EGFP-Fc fusion protein regarded as a report gene;which verified that the resistance of mutant-NPT to G418 was weakened.By comparing fluorescence assay of EGFP intensity in stable transfections after selection with the same concentration of G418 for 3 weeks;mutant-selected pools expressed more exogenous protein than the WT-selected pools.Therefore;the ratio of high producers in a transfected cell population greatly increased.

Simultaneous Determination of Neomycin Sulfate and Hydrochloric Dyclonine in Compound Twain Oint-ments by HPLC-DAD-ELSD / 中国药师

Objective:To establish an HPLC-DAD-ELSD method for the simultaneous determination of neomycin sulfate and hy-drochloric dyclonine in compound Twaln ointments. Methods:The assay was performed on an Agilent ZOR BAXSB-C18 column(250 mm × 4. 6 mm, 5μm) with acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1. 0 ml·min-1 . The detection wavelength of DAD was 282 nm. The evaporator temperature of ELSD was set at 50℃ and the nebulizer temperature was set at 60℃with the gas flow rate of 1. 6 L·min-1 . The column temperature was kept at 35℃. Results:The linear range of neomycin sulfate was 141. 54-323. 52 μg·mL-1(r=0. 999 6) with the average recovery of 98. 87%(RSD=0. 95%, n=9). The linear range of hydro-chloric dyclonine was 28. 00-64. 00 μg·mL-1(r=0. 999 6) with the average recovery of 99. 57%(RSD=1. 10%, n=9). Conclu-sion:The method is accurate, sensitive and reproducible, and under the same chromatographic conditions, the determination of all the active ingredients in compound Twaln ointments is achieved, which provides basis for the quality control.

Evaluation of Efficacy, Safety and Tolerability of a Combination of Clobetasol, Neomycin and Miconazole Versus Betamethasone, Clotrimazole and Neomycin Versus Betamethasone, Gentamycin and Miconazole in Subjects with Eczematous Disorders Associated with underlying Tinea or Yeast Infections.

Lobate GM Neo, 15 mg is a triple drug combination of a steroid clobetasol with anti-fungal miconazole and antibacterial neomycin in treatment of Eczematous disorders associated with underlying Tinea or Yeast Infections. Aims and Objectives: The study was designed to evaluate the efficacy, safety and tolerability of a combinations of clobetasol, neomycin and miconazole (Group A) versus betamethasone, clotrimazole, neomycin (Group B) versus betamethasone, gentamicin, miconazole (Group C) in subjects with any type of eczematous disorder associated with underlying tinea or yeast infection. Materials and Methods: This was an open label, parallel group, randomized comparative study. The primary endpoint analyzed was improvement in clinical score from baseline at the end of day 7 and other primary endpoint like hyperpigmentation were analyzed by the visual analogue scale of 1 to 10 at the end of day 7. Results: Thirty-six subjects were randomized to three groups. The clinical score showed a significant reduction from baseline at the end of day 7 in all the groups, i.e. 82.9%, 81.3% and 85.6% in Group A, B and C respectively. However, the difference between the groups were not statistically significant. Mean hyper pigmentation score showed significant decrease of 82.9% in Group A, 81.6% in Group B and 92.2% in Group C from baseline at the end of day 7. Conclusion: The triple combination of antifungal, antibacterial and potent steroid was found to be efficacious, safe and tolerable in reducing signs and symptoms (scaling, inflammation, burning and itching) of eczematous disorder associated with underlying tinea/yeast infection.

Eficacia, tolerancia y seguridad de una combinación de metronidazol, miconazol, centella asiática, polimixina y neomicina en cápsula blanda para uso vaginal en el tratamiento de la vaginosis bacteriana / Efficacy, tolerability, and safety of a combination including metronidazole, miconazole, Gotu kola, polymixin, and neomycin in soft vaginal capsules for the treatment of bacterial vaginosis

Introducción: La vaginosis bacteriana (VB) es un síndrome polimicrobiano, en la cual la flora dominante de lactobacilos normales es sustituida por una flora polimicrobiana. La prevalencia de VB en Perú varía entre 27 y 43,7%. El Centro de Control y Prevención de Enfermedades (DCD) sugiere el tratamiento de VB en mujeres sintomáticas con metronidazol oral/gel o clindamicina crema. Se planteó en el presente estudio evaluar la eficacia, tolerancia y seguridad de la combinación de metronidazol, miconazol, centella asiática, polimixina y neomicina en cápsula blanda para el tratamiento de VB. Material y Métodos: El presente estudio de tipo abierto, observacional, prospectivo, permitió evaluar la eficacia, tolerancia y seguridad en la aplicación de la combinación de metronidazol, miconazol, centella asiática, polimixina y neomicina en cápsula blanda. Resultados: Se incluyó a 61 pacientes con edad promedio de 29.28 años (rango 18-48) de las cuales 93,4% tenía historia previa de flujo vaginal anormal. Se realizaron dos visitas durante el estudio, la primera para diagnóstico e inicio de tratamiento y la segunda de control post tratamiento. Tres pacientes no tuvieron segunda visita y 8 no tenían registrada toda la información para definir la respuesta terapéutica. La segunda visita se realizó a los 21 días en promedio. Los principales signos y síntomas en la primera visita de diagnóstico fueron flujo vaginal (100,0%), disconfort vaginal (85,2%), dispareunia (70,5%) y dolor abdominal bajo (57,4%), las cuales disminuyeron en forma significativa (p<0,05) a la segunda visita post tratamiento. La prueba de aminas resultó positiva en el 93,4% de los casos en la primera visita y en el 15,5% de los casos en la segunda visita (p<0,05). De la población inicial de estudio, solo 53 mujeres son evaluables para eficacia terapéutica...

Introduction: Bacterial vaginosis (BV) is a polymicrobial syndrome, in which the normal dominant flora consisting in Lactobacillus is replaced by polymicrobial flora. The prevalence of BV in Peru varies between 27 and 43.7%. The Centers for Disease Control and Prevention suggest therapy for BV in symptomatic women should include oral/gel metronidazole or clindamycin cream. We proposed in this study to evaluate the efficacy, tolerability and safety of the combination of metronidazole, miconazole, Gotu kola (Centella asiatica), polymixin, and neomycin in soft capsules, for the treatment of BV. Material and Methods: This investigation was an open, observational, and prospective study, which allowed us to evaluate the efficacy, tolerability and safety of the aforementioned combined therapy administered in soft capsules. Results: The study included 61 patients with a mean age of 29.28 years (range, 18-48) and 93.4% had a history of abnormal vaginal discharge. Two visits took place during the study, the first for making the diagnosis and initiating therapy, and the second was the post-treatment control. Three patients did not have a second visit and 8 did not record all the information required to define the therapeutic response. The second visit took place after 21 days on average. The main signs and symptoms at the first visit were vaginal discharge at diagnosis (100.0%), vaginal discomfort (85.2%), dyspareunia (70.5%) and lower abdominal pain (57.4%), which were significantly reduced (p <0.05) in the second visit after treatment. The amine test was positive in 93.4% of cases in the first visit and in 15.5% of cases in the second visit (p <0.05). From the initial population in the study, only 53 women are evaluable for efficacy. An overall response rate in 44 women (83.02%) was achieved with the soft capsule combination treatment. Adverse events were reported in only one case...

Protective Role of Trimetazidine Against Neomycin-induced Hair Cell Damage in Zebrafish

OBJECTIVES: Trimetazidine (TMZ) is known to reduce the generation of oxygen-derived free radicals. The objective of the present study was to evaluate the effects of TMZ on neomycin-induced ototoxicity in transgenic zebrafish (Brn3C: EGFP). METHODS: Five-day, postfertilization zebrafish larvae were exposed to 125 microM neomycin and one of the following TMZ concentrations for 1 hour: 10 microM, 100 microM, 500 microM, 1,000 microM, 1,500 microM, or 2,000 microM. Hair cells within the neuromasts of the supraorbital (SO1 and SO2), otic (O1), and occipital (OC1) lateral lines were analyzed using fluorescence microscopy and confocal microscopy (n=10). Hair cell survival was calculated as a percentage of hair cells in the control group that were not exposed to neomycin. Ultrastructural changes were evaluated using scanning electron microscopy. RESULTS: TMZ protected against neomycin-induced hair cell loss in the neuromasts (TMZ 1,000 microM, 11.2+/-0.4 cells; 125 microM neomycin only, 4.2+/-0.5 cells; n=10; P<0.05) and decreased the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) reaction. In the ultrastructural analysis, structures of mitochondria and hair cells within the neuromasts were preserved in zebrafish exposed to 125 microM neomycin and 1,000 microM TMZ. CONCLUSION: TMZ attenuated neomycin-induced hair cell loss in zebrafish. The results of this study suggest that neomycin induces apoptosis, and that apoptotic cell death can be prevented by treatment with tremetazidine.

Antibacterial Mechanisms of Berberine and Reasons for Little Resistance of Bacteria / 中草药·英文版

Objective To study the antibacterial mechanisms of berberine and try to understand the reasons why bacteria cells difficultly resisted to it. Methods Detecting the minimal inhibitory concentration (MIC) of bacterial cultures incubated under sub-MIC concentration of berberine, Huanglian, and Neomycin for more than 200 generations, in order to analyze the bacteria resistance. Detecting the binding kinetics of berberine to DNA, RNA, and proteins. Observing the changes in bacterial cell surface structure with scanning electron microscopy. Detecting the Ca2+ and K.+ released from berberine-treated bacterial cells with atomic absorption spectrum. Detection the absorption of methyl-3H-thymine (3H-dT), 3H-uridine (3H-U), and 3H-tyrosine (3H-Tyr) into berberine-treated bacterial cells. Results MICs of bacterial cultures, growing more than 200 generations in MH medium with 1/2 MIC of berberine (BA200) or Huanglian (HA200), did not increase compared to the control, while remarkably increased in MH medium with 1/2 MIC of Neomycin (NA200). In addition, from the culture NA200 it was easy to isolate resistant mutant strains which could grow in MH medium with more than four times MIC Neomycin, but from the culture BA200 and HA200 it was difficult to isolate berberine or Huanglian mutant strains could grow in MH medium with more than four times MIC berberine or Huanglian. The binding kinetics of berberine to DNA, RNA, and proteins illustrated that berberine could easily and tightly bind to DNA and RNA, and hardly dis-bind from DNA- and RNA-berberine complexes. Berberine could easily bind to protein too, but also easily dis-bind from berberine-protein complex. The bacterial cells treated with berberine sharply decreased the absorption of 3H-dT, 3H-U, and 3H-Tyr, as the radioactive precursors of DNA, RNA, and protein biosynthesis. Berberine could damage bacterial cell surface structure, especially for Gram-negative bacteria. Ca2+ and K+ released from berberine-treated cells increased significantly compared to the control. Conclusion All of above results indicate that bacterial cells could not easily become resistant mutants to berberine. The mechanisms for the bactericidal effect of berberine include: inhibiting DNA duplication, RNA transcription, and protein biosynthesis; influencing or inhibiting enzyme activities; destructing the bacterial cell surface structure and resulting in Ca2+ and K+ released from cells. All of the berberine bactericidal mechanisms are the most essential physiological functions for a live cell, if influenced any one such function, the mutation would be lethal mutation, so that it is difficult to get berberine resistant cells. The results in this paper also prefigure that berberine and its related Chinese medicines would provide a feasible way to control antibiotic resistance problem.

A eritromicina utilizada como tratamento da encefalopatia hepática em portadores de cirrose: estudo prospectivo, randomizado, duplo-cego e controlado / Erythromycin used as treatment of hepatic encephalopathy in patients with cirrhosis: prospective study, randomized, double-blind and controlled

A encefalopatia hepática (EH) é uma complicação da cirrose considerada como sinal de mau prognóstico na doença hepática avançada. As conseqüências da EH não são completamente reversíveis, por isso o tratamento deve ter rápido início de ação e ser altamente eficaz. Atualmente os antibióticos são os medicamentos mais eficazes na EH aguda, porém no tratamento da EH são utilizados apenas os de baixa absorção, que em geral demandam certo tempo para apresentar resultados. Além disso, esses antibióticos não são isentos de efeitos adversos. Análise comparativa da eficácia da eritromicina (ERY), usada pela primeira vez no tratamento da EH em portadores de cirrose. Ensaio clínico prospectivo, randomizado e duplo cego de pacientes adultos portadores de cirrose, internados por EH no período de agosto/2008 a outubro/2010 no Hospital das Clínicas da Faculdade de Medicina de Botucatu - UNESP. Após a admissão, os indivíduos receberam tratamento com ERY 250 mg ou neomicina (NEO) 1 grama. As medicações foram utilizadas por via oral e administradas de 6/6 horas até a alta hospitalar, prescrição de outro antibiótico ou óbito. Todos os pacientes foram avaliados diariamente por meio do índice de encefalopatia hepática (IEH). Foram realizadas dosagens diárias de amônia sérica e proteína C reativa (PCR), bem como exames de bioquímica hepática e renal. A análise estatística foi realizada por meio de testes de comparação entre os grupos (testes t de Student e Mann-Whitney), medidas de distribuição central e análise descritiva. 30 casos de EH foram avaliados (15 tratados com ERY e 15 com NEO). Os grupos foram homogêneos em relação a idade, IEH, grau de EH à admissão hospitalar, escala de coma de Glasgow, escore Meld e classificação de Child-Pugh modificada.(...)

Hepatic encephalopathy (HE) is one of the major complications in patients with hepatic cirrhosis, and is considered a sign of bad prognosis in this setting. The consequences of HE are not totally reversible, thus the treatment must to be effective in a short time to attain HE regression. Actually, antibiotics are the best options in acute HE treatment. However, the antibiotics used in HE are drugs of low absorption, which often spend a long time to obtain its effect. Nevertheless, these antibiotics are not free of adverse effects. Comparative analysis of efficacy of erythromycin (ERY) used by the first time as a treatment of HE. Randomized controlled trial of adult patients with HE and hepatic cirrhosis admitted from August 2008 to October 2010 in the hospital of the Botucatu Medical School - UNESP. After randomization, the patients received either ERY 250 mg or neomycin (NEO) 1 g. The drugs were administered orally q.i.d. until hospital discharge, prescription of another antibiotic or death. All subjects were evaluated diary and the hepatic encephalopathy index (HEI) was calculated every day. Serum ammonia, C reactive protein (CRP) and biochemical profile (hepatic and renal exams) were obtained diary of each patient. Statistical analysis was performed using tests for comparison between the groups (Student t test and Mann-Whitney test), Spearman’s rank correlation, central distribution measures and descriptive analysis. 30 cases of HE were evaluated (15 treated with each drug). In the moment of admission, the groups were homogeneous with respect to age, HEI, HE grade, Glasgow coma scale, Meld score and Child-Pugh modified classification. The subjects that received ERY had a short time of hospitalization (p = 0.032) and a great degree in the alanine aminotransferase (ALT) levels (p = 0.026).(...)

Spatiotemporal dynamics of pharmacological modulation of membrane PtdIns(4,5)P2 metabolism by different agents / 中国药理学通报

Aim: To visualize the dynamics of PtdIns (4,5) P2 hydrolysis and resynthesis, and modulate it by pharmacological agents wortmannin, LiCl, U73122 and neomycin. Methods: We used a fusion construct of green fluorescent protein (GFP) with the PH domain of phospholipase Cδ1 (PLC δ1 PH) (PLCδ1 PH-GFP) known to bind PtdIns(4,5) P2 specifically, and laser-scanning confocal microscopy to trace PtdIns (4,5) P2 translocation. Results: Stimulation of endogenous P2Y receptors by ATP in CHO cells induced a reversible PLCδ1 PH-GFP translocation, indicating PtdIns (4,5) P 2 hydrolysis through the receptor-mediated phospholipase C (PLC) activation. Wortmannin and LiCl did not affect the translocation of PLC δ1 PH-GFP from plasma membrane to cytosol but blocked the recovery after the translocation. The transient translocation from plasma membrane was blocked by the PLC inhibitor U73122 but was not affected by another PLC inhibitor neomycin. However, in the absence of PLCδ1 PH-GFP expression, neomycin inlibited the receptor-induced PLC hydrolysis of PtdIns(4,5) P2. Conclusion: PLCδ1 PH-GFP can be used as a valuable fluorescence probe to visualize the dynamic change of PtdIns (4,5) P2 in living cells. Wortmannin, LiCl, U73122 and neomycin have distinct modulation effects on PtdIns(4,5) P2 metabolism. PLC δ1 PH, when bound to PtdIns(4,5) P2, prevents neomycin from inhibiting PLC hydrolyzing PtdIns(4,5) P2.

The induction and elimination of bacteria's resistance / 中国药理学通报

AIM Objective By comparing E.coli's resistance to antibiotics and Chinese medical herbs before and after incubated in LB containing Neomycin or Chinese medical herbs, we try to understand the difference in their mechanism of resistance. METHODS We incubated E.coli cells under the culturing media with low concentration of Neomycin and Chinese medicine herbs respectively; lately, the culturing process were continued under the media with no Neomycin or Chinese medicine herbs. We tested the minimal inhibitory concentrations(MIC)of Neomycin(Kanamycin, Gentamicin, Tetracyclin etc) and Chinese medical herbs to culture cells. Result The experimental data indicated that increased resistance of population cells in the broth including Neomycin happened easily, as well as multi resistance simultaneously. As for the Chinese medical herbs, the case is different. After growing in the LB containing one of Huang Lian (rhizoma coptidis), Berberine or San-Huang-Tang of low concentrations, population cells did not increase their resistance either to antibiotics or to Chinese medical herbs. Removing the pressure of Neomycin, E.coli population cells resistance came back to the initial level. CONCLUSION The increased antibiotic resistance of population cells is unstable, and alternative using of different antibiotics maybe contribute to the alleviation of antibiotics resistance.