Experimental study of proteasome inhibitor MG132 up-regulates Wnt/β-catenin signaling pathway to improve osteoporosis / 中国骨伤

OBJECTIVE@#To explore the mechanism of proteasome inhibitor MG132 in improving osteoporosis.@*METHODS@#Total of 32 female SD rats, weighing 220 to 250 g and 8 weeks old, were selected. They were randomly divided into 4 groups(n=8). Rats of group A and group B were cut off ovaris on both sides to make model of osteoporosis, and then they were given proteasome inhibitors MG132 and dimethyl sufoxide (DMSO) respectively. Group C was a sham group and rats were given MG132. Group D was a normal group and rats were given MG132 too. The rats were killed in batches at 6 and 12 weeks after administration, and the femoral neck tissues were obtained. Relevant data were analyzed, such as pathomorphological observation, micro-CT analysis, detection of 20S proteasome activity in tissues, and expression of Wnt and β-catenin.@*RESULTS@#Morphological observation showed that the trabecular were slightly thinner, reticulated, and occasionally interrupted in group A, while the trabecular were obviously thinner and discontinuous in group B. And the trabecular were intact and arranged reticulated in group C and D. The analysis results of bone mineral density(BMD), bone surface(BS), bone volume/total volume(BV/TV) and trabecular thickness(Tb.Th) showed that group B was worse than other groups in all parameters at different time points(P<0.05), and group A was worse than group C and group D in BS(P<0.05), there was no significant difference in all parameters between group C and group D. RFU value of 20S proteasome in group B was significantly higher than that in other groups(P<0.05). According to the results of Western blot, the gray values of Wnt protein and β-catenin protein in group A were significantly higher than those in other groups (P<0.05).@*CONCLUSION@#MG-132, a ubiquitin proteasome inhibitor, can regulate Wnt/β-catenin signaling pathway by inhibiting the degradation of β-catenin protein, and delaying the occurrence and development of osteoporosis.

FilmArray Meningitis/Encephalitis (ME) panel in the diagnosis of bacterial meningitis

ABSTRACT The precise diagnosis of bacterial meningitis is essential. Cytological and biochemical examination of cerebrospinal fluid (CSF) are not specific. Conventional methods for bacterial meningitis lack sensitivity or take too long for a final result. Therefore, other methods for rapid and accurate diagnosis of central nervous system infections are required. FilmArray meningitis/encephalitis (ME) panel is a PCR multiplex for simultaneous and rapid identification of 14 pathogens, including 6 bacteria, 7 viruses, and Cryptococcus. We evaluated 436 CSF samples submitted to FilmArray ME Panel. Among them, 25 cases were positive for bacteria, being Streptococcus pneumonia the most frequent (48 %). Among positive cases for bacteria, 60 % were positive only with FilmArray. All the bacterial meningitis cases in which the only positive test was FilmArray had CSF findings suggestive of bacterial meningitis, including neutrophilic pleocytosis, increased CSF protein and lactate, and decreased CSF glucose. These findings suggest that FilmArray may increase the diagnostic sensitivity for bacterial meningitis.

Effectiveness of over-the-counter tooth-whitening strips and toothbrushing for stain removal / Eficácia da utilização de tiras de clareamento dental e da escovação na remoção de manchas dentais

Objective: The present study compared the effectiveness of over-the-counter tooth-whitening strips and too-thbrushing for stain removal. Materials and method: Forty bovine teeth were subjected to initial color re-adings and divided into two groups: 20 teeth were stained with grape juice for seven days and 20 did not receive any treatment (control). Each group was subdivided into two groups (n=10) according to treatment: whitening strips twice/day (30 minutes/7 days) and mechanical brushing with toothpaste (one year). After the treatments, final color readings were taken and color stability (ΔEab), coordinates (ΔL, Δa, Δb), and whitening index (WID) were calculated and analyzed statistically (two-way ANOVA, Bonferroni α=0.05). Results: Statis-tical differences were found for ΔEab and all coordinates between the Control and Stained groups, with major changes in the Stained groups for both treatments. In the Control group, whitening strips produced greater changes in Δb (p <0.05), indicating decrease of yellowness. For the Stained group, whitening strips produced greater changes in Δa (p <0.05), indicating redness reduction. Conclusion: Both treatments were effective for removing stains, and whitening strips were more efficient and effective for changing the color of teeth without previous staining. (AU)

Objetivo: o presente estudo comparou a eficácia das tiras de clareamento dentário e da escovação na remoção de manchas. Materiais e método: quarenta dentes bovinos foram submetidos à lei-tura inicial de cor e divididos em dois grupos: 20 manchados com suco de uva por 7 dias e 20 não receberam tratamento (controle). Cada grupo foi subdividido em 2 grupos (n=10) de acordo com o tratamento: tiras de clareamento 2 vezes/dia (30 minutos/7 dias) e escovação mecânica com cre-me dental (1 ano). Após os tratamentos, leituras finais de cor foram realizadas. Então, a estabili-dade de cor (ΔEab), as coordenadas (ΔL, Δa, Δb) e o índice de clareamento (WID) foram calculados e analisados estatisticamente (two-way ANOVA, Bonferroni α=0,05). Resultados: diferenças es-tatísticas foram encontradas em ΔEab e todas as coordenadas entre os grupos controle e mancha-dos, com maior alteração no grupo manchado, independente do tratamento. No grupo controle, o clareamento produziu maior alteração em Δb (p <0,05), indicando redução do croma amarelo. Nos dentes manchados, ocorreu maior alteração em Δa nos dentes clareados (p <0,05), indicando diminuição do croma vermelho. Conclusão: am-bos os tratamentos foram eficazes na remoção de manchas e o clareamento dentário mostrou-se um método mais eficiente e efetivo para alteração de cor de dentes que não haviam sido submetidos à pigmentação prévia. (AU)

Relationship between p53 rs1625895 polymorphism and prognosis in diffuse large B-cell lymphoma / 北京大学学报(医学版)

OBJECTIVE@#p53 gene, as "the guardian of the genome", is the most widely studied tumor suppressor gene. Previous studies have shown that about 50 percent of tumors have P53 dysfunction. This article aims to retrospectively analyze the correlation between p53 rs1625895 polymorphism and the prognosis of patients with diffuse large B-cell lymphoma (DLBCL).@*METHODS@#PCR combined with Sanger sequencing were used to detect rs1625895 genotype in 384 DLBCL patients. The relationship between rs1625895 polymorphisms and the clinical characteristics, first-line therapeutic effects and the prognosis of the patients were analyzed.@*RESULTS@#Among all the patients, 2 (0.5%) patients with AA genotype, 34 (8.9%) patients with AG genotype and 348 (90.6%) patients with GG genotype were identified. The patients with different rs1625895 genotypes did not have any difference in terms of age, gender, B symptoms (developing any of the following symptoms: unexplained recurrent fever (often above 38 °C), night sweats, and unexplained weight loss of 10% within 6 months ), erythrocyte sedimentation rate (ESR), international prognostic index (IPI) and molecular subtype (P>0.05). The overall response rate (ORR) was 82.9% and 82.8% in AA/AG and GG, respectively. There was no significant difference between the first-line therapeutic effects of the two groups (P>0.05). And there was also no difference between A allele carriers and homozygous G allele carriers for the 5-year progressionfree survival rate (PFS) (71.8% vs. 62.3%, χ2=1.351, P=0.245) and 5-year overall survival rate (OS) (72.2% vs. 64.1%, χ2=1.267, P=0.260). But in the subgroup with Germinal Center B-cell (GCB) type, the patients carrying A allele for rs1625895 had an obviously longer PFS (91.7% vs. 72.7%, χ2=4.493, P=0.034) and OS (91.7% vs. 76.7%, χ2=4.246, P=0.039) compared with the patients homozygous for the G allele. As for the patients with non-GCB subtype, there was no significant difference in PFS and OS between different rs1625895 genotypes (P>0.05). According to whether the first-line regimen contained rituximab or not, the patients were divided into two groups treated with cyclophosphoramide, doxorubicin, vincristine and prednisone (CHOP) or with rituximab and CHOP (R-CHOP). But in both subgroups, there was no significant difference in the 5-year PFS and OS between the AA/AG and GG patients, too (P>0.05).@*CONCLUSION@#For DLBCL patients receiving CHOP regimen chemotherapy in the first line, p53 rs1625895 cannot predict the clinical efficacy and prognosis of the patients, but in the patients with GCB subtype, this polymorphism may be a prognostic indicator.

O mercado brasileiro de anticorpos monoclonais utilizados para o tratamento de câncer / The Brazilian market for monoclonal antibodies used in cancer treatment / El mercado brasileño de anticuerpos monoclonales utilizados para el tratamiento de cáncer

Resumo: Os anticorpos monoclonais (mABs) têm sido indicados como tecnologia inovadora para o tratamento de alguns tipos de câncer, por serem capazes de alvejar e matar seletivamente células tumorais. Contudo, os altos custos dessas terapias colocam em questão a sustentabilidade do acesso. Este trabalho teve como objetivo identificar as principais características dos anticorpos monoclonais, destinados ao tratamento de câncer, com registro sanitário ativo, no Brasil, em 2016. Tratou-se de uma análise descritiva retrospectiva a partir de consulta à página de Internet da Agência Nacional de Vigilância Sanitária (Anvisa), em que esses mABs foram caracterizados de acordo com antígeno-alvo, tipo de anticorpo, ano de registro, indicações terapêuticas e empresa detentora do registro. Foram identificados 14 anticorpos com ação em sete antígenos-alvo diferentes. No que diz respeito às indicações clínicas, houve uma maior frequência de linfomas, leucemias, câncer de mama e câncer colorretal. Quanto ao tipo, foram identificados três anticorpos quiméricos, seis humanizados e cinco humanos. A Roche apareceu como a empresa detentora do registro de 6 dos 14 mABs, o que representa 43% dos registros sanitários. Foi possível, a partir desses dados, discutir a ideia de medicamentos me-too no mercado de biológicos, assim como pensar as tensões existentes nesse mercado e a ideia de oligopólio diferenciado. Apesar do desenvolvimento de novos produtos, ainda que para atuar em um mesmo alvo, representar a possibilidade de um incremento competitivo e, com isso, de uma diminuição dos preços praticados pelas empresas torna-se um problema quando é a mesma empresa que lança no mercado novos anticorpos direcionados ao mesmo alvo, sem mudanças relevantes.

Abstract: Monoclonal antibodies (mABs) have been indicated as an innovative technology for the treatment of some types of cancer, since they are capable of targeting and selectively killing tumor cells. However, the high costs of these therapies raise questions as to the sustainability of access. This study aimed to identify the principal characteristics of monoclonal antibodies used in cancer treatment with active marketing authorization in Brazil as of 2016. This was a descriptive retrospective analysis based on consultation of the Brazilian Health Regulatory Agency (Anvisa) website, in which these mABs were characterized according to the target antigen, type of antibody, year of registration, therapeutic indications, and applicant. A total of 14 antibodies were identified with action on seven different target antigens. The most frequent clinical indications were for lymphomas, leukemias, breast cancer, and colorectal cancer. As for type, the study identified three chimeric, six humanized, and five human antibodies. Roche was the applicant in 6 of the 14 mABs, or 43% of the marketing authorization. It was possible to discuss the idea of me-too medicines in the biological market and the idea of a differentiated oligopoly, as well as to think about the tensions in this kind of market. It is expected that the development of new products, although to act on the same biological target, represent the possibility of a competitive increase and, as a result, a decrease in prices practiced by companies. However, this becomes a problem when it is the same pharmaceutical industry that launches on the market new antibodies directed to the same target, with no relevant changes.

Resumen: Los anticuerpos monoclonales (mABs) han sido señalados como una tecnología innovadora para el tratamiento de algunos tipos de cáncer, por ser capaces de apuntar y matar selectivamente células tumorales. No obstante, los altos costes de estas terapias ponen en cuestión la sostenibilidad del acceso. El objetivo de este trabajo fue identificar las principales características de los anticuerpos monoclonales, destinados al tratamiento de cáncer, con registro sanitario activo, en Brasil, en 2016. Se trató de un análisis descriptivo retrospectivo, a partir de la consulta a la página web de la Agencia Nacional de Vigilancia Sanitaria (Anvisa), donde esos mABs se caracterizaron conforme el antígeno objetivo, tipo de anticuerpo, año de registro, indicaciones terapéuticas y empresa detentora de su registro. Se identificaron 14 anticuerpos con acción en siete antígenos-objetivo diferentes. En lo referente a las indicaciones clínicas, hubo una mayor frecuencia de linfomas, leucemias, cáncer de mama y cáncer colorrectal. En cuanto al tipo, se identificaron tres anticuerpos quiméricos, seis humanizados y cinco humanos. Roche apareció como la empresa detentora del registro de 6 de los 14 mABS, lo que representa un 43% de los registros sanitarios. Fue posible, a partir de esos datos, discutir la idea de medicamentos me-too en el mercado de biológicos, así como reflexionar sobre las tensiones existentes en ese mercado y la idea de oligopolio diferenciado. El desarrollo de nuevos productos, aunque sean para actuar en un mismo objetivo, representa la posibilidad de un incremento competitivo y, con ello, de una disminución de los precios practicados por las empresas. Esto se convierte en un problema cuando es la misma empresa que lanza en el mercado nuevos anticuerpos, dirigidos al mismo objetivo, sin cambios relevantes.

Relationship of ecosystems and resolution of new drug research and development / 中国药学杂志

OBJECTIVE: Relationship of ecosystems and resolution of new drug research and development is analyzed through a comparison of the clinical needs, industry development policy, enterprise-self factor, listing application mechanism, number and characteristics of new drug in America and China, especially, the market conditions of the me-only, me-better and me-too between America and China is analyzed importantly. The idea that the new drug is studied and developed in China is presented. METHODS: The important influencing factors that affected the ecosystems of new drug research and development were analyzed by comparison METHODS and data studies. RESULTS: The clinical availability of new drug in America is significant increased recently and over 90% applied new drugs can be finished in a limited time based on the evolution of the innovative ecological environment of new drug research and development from 1993 to 2014, new drugs are approval listing in America and China respectively. CONCLUSION: The availability of new drug is very important to a state security. The ecosystem of new drug research and development is very complicated. The relative field in America is more advanced than in China and it is used for reference to the new drug research and development in China.

Comparative Study of First-in-Human Dose Estimation Approaches using Pharmacometrics

OBJECTIVE: First-in-human dose estimation is an essential approach for successful clinical trials for drug development. In this study, we systematically compared first-in-human dose and human pharmacokinetic parameter estimation approaches. METHODS: First-in-human dose estimation approaches divided into similar drug comparison approaches, regulatory guidance based approaches, and pharmacokinetic based approaches. Human clearance, volume of distribution and bioavailability were classified for human pharmacokinetic parameter estimation approaches. RESULTS: Similar drug comparison approaches is simple and appropriate me-too drug. Regulatory guidance based approaches is recommended from US Food and Drug Administration (FDA) and European Medicines Agency (EMA) regarding no-observed-adverse-effect level (NOAEL) or minimum anticipated biological effect level (MABEL). Pharmacokinetic based approaches are 8 approaches for human clearance estimation, 5 approaches for human volume of distribution, and 4 approaches for human bioavailability. CONCLUSION: This study introduced and compared all methods for first-in-human dose estimation. It would be useful practically to estimate first-in-human dose for drug development.

The perceptions of medical researchers on qualitative methodologies / Percepción de los investigadores médicos en metodologías cualitativas / Percepção de pesquisadores médicos sobre metodologias qualitativas

We aimed to verify doctor's perception of the qualitative research method, via a qualitative study of interviews with questions on the academic profile of doctors and on the methodology. We interviewed 42 professionals, of which 18 had experience with the qualitative method and 24 with the quantitative method. The results showed that knowledge on the qualitative method was virtually nil among "quantitative researchers", who did not value qualitative research, although some of those realized that it would be important to be more accepting in clinical practice. Others only considered the method as subsidiary to quantitative. The majority considered qualitative methods as lacking academic structure, taking too long to conduct empirical studies, and being difficult to publish. All of them criticized the misuse of the method, and the "quantitatives" pointed out the problem of being unable to reproduce. We concluded that widening the use of the qualitative method by doctors requires investment from the beginning of the academic career and participation in qualitative research projects.

El objetivo es verificar la percepción de médicos sobre el método de investigación cualitativa. Se trata de un estudio cualitativo por medio de entrevistas con preguntas sobre el perfil de los médicos y sobre el método. Entrevistamos a 42 profesionales, 18 con experiencia en el método cualitativo y 24 con el cuantitativo. Los resultados mostraron que el conocimiento sobre lo cualitativo es casi nulo entre los "cuantitativistas", que no valoran la investigación cualitativa, aunque algunos se dan cuenta de que sería importante tener un enfoque más amplio en la práctica clínica. Otros la ven como subsidiaria a lo cuantitativo. Sus dificultades para utilizar ese abordaje son: falta de formación, cantidad de tiempo que exigen y problemas de publicación. Todos han criticado el mal uso del método. Los "cuantitativistas" han destacado como fragilidad, la no reproductibilidad. Llegamos a la conclusión de que para ampliar el uso de los abordajes cualitativos entre los médicos es importante invertir en su formación desde el inicio del curso y la participación en proyectos de investigación cualitativa.

Objetivamos verificar a percepção de médicos sobre o método qualitativo de pesquisa. Estudo qualitativo por meio de entrevistas com questões sobre o perfil acadêmico do médico e perguntas abertas a respeito do método. Entrevistamos 42 profissionais, sendo 18 com experiência no método qualitativo e 24 com o quantitativo. Os resultados evidenciaram que o conhecimento sobre o qualitativo é quase nulo entre os pesquisadores "quantitativistas", os quais não valorizam a pesquisa qualitativa, embora alguns percebam que seria importante ter uma postura mais compreensiva na prática clínica. Outros só a veem como subsidiária ao quantitativo. As principais dificuldades da maioria são: falta de formação, tempo longo despendido nos estudos empíricos e dificuldade de publicação. Todos os entrevistados criticaram o mau uso do método, e os "quantitativistas" ressaltaram, como problema, sua não reprodutibilidade. Concluímos que ampliar o uso do método qualitativo por médicos exige investimento na formação desde o início da graduação e participação em projetos de pesquisa qualitativa.

Industry withdrawal from psychiatric medication development

Between 1950 and 1969, on a serendipitous basis, psychiatric drug development flourished. However, there has been a steep decline in the development of new medication classes. Instead of new molecular entities, slight molecular modifications producing “me-too” drugs attempted to garner market share. With failing profitability, industry is now withdrawing from psychiatric medication development. Managed care drastically shortened contact between patients and clinicians, so the possible observation of unexpected benefits has been nullified. The randomized, parallel-groups design met FDA requirements for specific pharmacological efficacy. However, it does not determine whether a patient who improved while drug-treated required the drug or would have gotten better on his own. Further, pathophysiology benefit remains obscure. The major psychotropic drugs have no benefits for normal subjects. Their remarkable benefits must stem from a necessary interaction with a pathophysiological state. Therefore, understanding therapeutic benefit by treating normal subjects becomes unlikely. The claim that therapeutic knowledge in psychiatry proceeds from bench to bedside has proven vacuous, primarily because of our limited understanding of brain pathophysiology. The utility of the alternative intensive design for understanding diagnosis, therapeutic benefit, and pathophysiology is emphasized.

Validation of high-performance liquid chromatographic method for analysis of fluconazole in microemulsions and liquid crystals

In recent decades, there has been a significant increase in the incidence of fungal diseases. Certain fungal diseases cause cutaneous lesions and in the usual treatment, generally administred orally, the drug reaches the site of action with difficulty and its concentration is too low. An approach much explored in recent years is the development of nanotechnology-based drug delivery systems, and microemulsions (ME) and liquid crystals (LC) are promising. ME and LC were developed with oleic acid or copaiba oil as the oil phase, propoxyl (5OP) ethoxyl (20 OE) cetyl alcohol as surfactant and water. An analytical method to assess the incorporation of fluconazole (FLU) in the systems under study was validated according to guidelines of the International Conference on Harmonization (ICH) guidelines and the Brazilian Food, Drug and Sanitation Agency (ANVISA). The method was conducted on a C18-RP column (250 × 4.6 mm i.d.), maintained at room temperature. The mobile phase consisted of acetonitrile and water (50:50, v/v), run at a flow rate of 1.0mL/min and using ultraviolet detection at 210nm. The chromatographic separation was obtained with a retention time of 6.3min, and was linear in the range of 20-400 µg/mL (r2=0.9999). The specificity showed no interference of the excipients. The accuracy was 100.76%. The limits of detection and quantitation were 0.057 and 0.172 µg.mL-1, respectively. Moreover, method validation demonstrated satisfactory results for precision and robustness. The proposed method was applied for the analysis of the incorporation of FLU in ME and LC, contributing to improve the quality control and to assure the therapeutic efficacy.

Nas últimas décadas, houve aumento significativo na incidência de doenças fúngicas. Certas doenças fúngicas provocam lesões cutâneas, sendo que no tratamento usual, geralmente administrado por via oral, o medicamento chega ao local de ação com dificuldade, em concentração muito baixa. Uma abordagem muito explorada nos últimos anos é o desenvolvimento de sistemas de administração de fármacos baseados em nanotecnologia, como as microemulsões (ME) e cristais líquidos (LC). ME e LC foram desenvolvidos com o ácido oleico ou óleo de copaíba como fase oleosa, álcool cetílico propoxilado (5 OP) e etoxilado (20 OE) como tensoativo e água. Método analítico para avaliar a incorporação de fluconazol (FLU) nos sistemas em estudo foi validado de acordo com as diretrizes da Conferência Internacional de Harmonização (ICH) e Agência Nacional de Vigilância Sanitária (ANVISA). O método foi desenvolvido empregando coluna C18-RP (250 x 4,6 mm id), mantida à temperatura ambiente. A fase móvel consistiu de acetonitrila e água (50:50, v/v), executado a uma taxa de fluxo de 1,0 mL/min e com detecção ultravioleta a 210 nm. A separação cromatográfica foi obtida com o tempo de retenção de 6,3min, e mostrou-se linear no intervalo de 20-400 µg/mL (r2=0,9999). Pelo estudo de especificidade, observou-se não interferência dos excipientes. A precisão foi 100,76%. Os limites de detecção e de quantificação foram 0,057 e 0,172 µg.mL-1, respectivamente. Além disso, a validação do método demonstrou resultados satisfatórios para a precisão e robustez. O método proposto foi aplicado para a análise da incorporação do FLU em ME e cristais líquidos, contribuindo para aumentar o controle de qualidade e garantir a eficácia terapêutica.

Application of methyl in drug design / 药学学报

The methyl group plays an important role in the rational drug design. Introducing methyl into small molecules has become an important strategy of lead compound optimization. The application of methyl in drug design is reviewed in this paper. Methyl can modulate the physicochemical, pharmacodynamic, and pharmacokinetic properties by ortho effect, inductive effect, and conformational effect. It also improves the metabolic stability as a soft metabolic point. In addition, introducing methyl into drug molecules can also be applied as a strategy in new uses of old drugs and generate me-too drugs.

Application of methyl in drug design / 药学学报

The methyl group plays an important role in the rational drug design. Introducing methyl into small molecules has become an important strategy of lead compound optimization. The application of methyl in drug design is reviewed in this paper. Methyl can modulate the physicochemical, pharmacodynamic, and pharmacokinetic properties by ortho effect, inductive effect, and conformational effect. It also improves the metabolic stability as a soft metabolic point. In addition, introducing methyl into drug molecules can also be applied as a strategy in new uses of old drugs and generate me-too drugs.

Detection of metallo-β-lactamases producing Acinetobacter baumannii using microbiological assay, disc synergy test and PCR.

Background: One leading factor responsible for resistance in Acinetobacter baumannii, an important opportunist in health care institutions globally, is the production of carbapenamases like metallo-β-lactamases (MBLs), which hydrolyze a variety of β-lactams including penicillin, cephalosporins and carbapenems. However, neither any standard guidelines are available nor any method has been found to be perfect for their detection. Various methods have shown discordant results, depending upon the employed methodology, β-lactamase substrate and MBL inhibitor used. This study aims to evaluate two phenotypic methods against PCR as gold standard among carbapenem resistant A. baumannii for identifying MBL producers. Materials and Methods: A total of 130 A. baumannii were screened for imipenem and meropenem resistance by Kirby-Bauer disc diffusion method. Phenotypic expression of MBL was detected by EDTA-imipenem-microbiological (EIM) assay and extended EDTA disc synergy (eEDS) test and presence of bla-IMP and bla-VIM was detected by PCR in all the carbapenem resistant isolates. Results: Of the 43 imipenem and/or meropenem resistant A. baumannii isolates, 4 (9.3%) were found to be MBL producers by EIM and 3 (6.97%) by eEDS. Only bla-VIM gene was detected in 7 (16.28%) by PCR. In addition EIM detected 14 (32.56%) carbapenem resistant non-metallo enzyme producers. Conclusion: Of the two MBL genes targeted, bla-VIM was only detected and that too in isolates resistant to both imipenem and meropenem. Further, EIM was useful in differentiating MBL from non-metalloenzymes producers.

Psicofármacos en niños y adolescentes II: psicoestimulantes, estabilizadores del ánimo y benzodiacepinas. revisión / Psychopharmacology in children and adolescents II: psychostimulants, mood stabilizers and benzodiazepines. review

Psicoestimulantes (PSE), estabilizadores del ánimo (EA) y benzodiacepinas (BDZ) son psicofármacos frecuentemente utilizados en psiquiatría infantojuvenil y cuya indicación ha aumentado considerablemente en los últimos años. Los PSE son empleados principalmente en el déficit atencional, siendo la terapia de primera línea en pacientes sin comorbilidad. Modafinilo y anfetaminas serían igualmente efectivos y se utilizan con similar frecuencia en muchos países sin embargo existe evidencia de mayores riesgos con anfetaminas. Otros fármacos como atomoxetina, a-agonistas, bupropión y antidepresivos tricíclicos (imipramina) se utilizan como tratamiento de segunda o tercera línea. Los EA son especialmente útiles en el tratamiento del trastorno bipolar (TB) y se consideran agentes de primera línea: litio, ácido valproico (sodio divalproex) y algunos antipsicóticos atípicos. Otros EA que han demostrado efectividad en TB son lamotrigina, carbamazepina, oxcarbamazepina y topiramato. Las DBZ han sido poco estudiadas en población infantojuvenil, pero su uso no es infrecuente. Si bien no son una indicación de primera línea en trastornos de ansiedad, están indicadas como tratamiento adyuvante especialmente diazepam. Fluracepam tiene un rol en el tratamiento de insomnio en mayores de 15 años. Aunque el tratamiento de elección de muchas patologías de psiquiatría infantojuvenil es claro en diversas guías clínicas, la complejidad terapéuticas de estos pacientes muchas veces hace necesario el uso de psicofármacos de segunda o tercera línea, muchos de ellos no aprobados por la Food and Drug Administration. Es estos casos cobra vital importancia el seguimiento estricto, con especial cuidado en los efectos colaterales, que pueden ser nocivos para el desarrollo del paciente.

Psycho stimulants (PSE), mood stabilizers (ME) and benzodiazepines (BDZ) are psychotropic’s frequently used in children and adolescents psychiatry and their indication has increased considerably in the last year. The PSAE are used mainly in attentional deficits being first-line treatment in patients without co-morbidity. Modafinil and amphetamines are supposed to be equally effective and used with similar frequency in many countries, however there is evidence of bigger risk with amphetamines. Other drugs such as Atomoxetine, a-agonist, bupropion, tricyclic antidepressants (imipramine) are used as second or third line treatment. The ME are especially useful as treatment of bipolar disorder (BD); lithium, divalproex sodium and some atypical antipsychotics are considered first line treatment. Other ME that have showed effectiveness in TB are lamotrigine, carbamazepine, oxcarbamazepina and topiramate. The BDZ haven´t been studied too much in young population, however its uses not uncommon. Although it is not a first line indication in anxiety disorders they are indicated as adjunctive treatment, especially diazepam. Fluracepam has a role in the treatment of insomnia in children over 15 years. While the treatment of choice for many pathologies of juvenile psychiatry is clear in several clinical guidelines, the therapeutic complexity of these patients often requires the use of psychopharmacology of 2nd or 3rd line, many of them not approved by the Food and Drug Administration in these cases is of vital importance a strict monitoring, with particular attention to side effects, which can be harmful to the patients development.

[ effect of hydrochloric extraction of valerian on number and size of raphe magnus neurons in adult rats]

Effective materials from Valerian officinalis L. have too much usage in the pharmacological industry. It is used as a sedative, anticonvulsion, and antidepressant drug. Serotonin has a widespread role in vital function such as sleep, awareness and calmness. In this study we evaluated the effect of hydrochloric extract of valerian on number and size of raphe magnus neurons in adult rat. In this experimental study, which was conducted at Yasuj University of Medical Sciences in 2009, forty adult Wistar rats, each 170-250 gr, were divided randomly into four groups [one control group and three experimental groups]. The animals were injected daily for one month with doses of 300, 400 and 600 mg/kg of the extract. The control group just received distilled water. After transcardial perfusion, the whole brain was separated, then 10 micro m sections of the brain stem were prepared, and hematoxylin and eosin [H and E] staining were done. Number and size of raphe magna neurons were observed under light microscope. The gathered data were analyzed by the SPSS software using One-way ANOVA and LSD. The control group did not statistically show significant changes in number of raphe magna neurons. Comparison of the means of long and short diameter neurons showed significant increases in experimental groups with control group [P<0.05]. In experimental groups the neuron nucleuses were more euchromatic than the control group. Hydrochloric extract of valerian has no effect on raphe magnus neurons, but it is effective on neurons' size. It can be concluded that the extract increases both neurons activity and serotonin secretion

Potential Cost Savings from Generic Medicines - Protecting the Prescribed Minimum Benefits

"Background: South Africa has followed a pro-generic policy since the introduction of the National Drug Policy in 1996. The selection processes in the public and private sectors have; however; remained largely disconnected; and at times contradictory. Medicines provided outside of hospitals accounted for 17of medical aid spend in 2006; up 8.8from the previous year. Of particular concern to funders has been the expenditure on the 27 chronic conditions listed as Prescribed Minimum Benefits. The Medical Schemes Act (No 131 of 1998) provides for the definition of Prescribed Minimum Benefits; which stipulate a package of services or care a medical scheme must provide for in its benefit design. There is pressure to reconsider these requirements in order to increase the affordability of medical scheme coverage. This study assessed the potential savings thatwould be achievable by substituting generics for brand name (originator) medicines listed in the chronic disease algorithms set out by the Council for Medical Schemes (CMS).Methods: All medicines listed in the 25 chronic diseases algorithms made available by the CMS were identified. Brand and generic versions were identified in the Monthly Index of Medical Specialties (MIMS; May 2006). Single exit prices inclusive of value added tax were obtained from the web site of the Pharmaceutical Blue Book and the cost per defined daily dose for one month was then calculated. Cost differentials; where available; were then identified for each medicine listed in the algorithms. Cost differentials for medicines within each algorithm were presented as the median of the difference between brand and generic medicines listed for that algorithm; and also as the median of differences between generic medicines for the same condition. Results: Three of the algorithms (diabetes insipidus; haemophilia and hypothyroidism) list medicines for which no generic equivalent was available at the time of the study. The median cost differential between brand and generic equivalents for the remaining 22 chronic conditions ranged from 19.5(for type 1 diabetes mellitus) to 97(for Addison's disease). Across the entire chronic disease algorithm set; 80 medicines with generic equivalents were listed for 22 conditions. The median cost differential between brand and generic versions of these 80 medicines was 49.9(interquartile range 32.0 to 78.5). Of all generic medicines identified; 67.5were more than 40cheaper; per defined daily dose (DDD) per month; than the branded version. In 16 medicines the cost differentials between generic versions were 1or less. Some correlation between the number of generics and the size of the cost differential was apparent (correlation coefficient 0.49). There were examples of high-cost differentials in highly competitive areas of the market. Conclusions: An argument could be made for more closely aligning the process of developing the National Essential Drugs List and the development of the CMS algorithms. By being more specific about which medicines should be covered; needless expenditure on ""me-too"" agents of doubtful additional benefit could be avoided. Where clinically warranted; appropriate choices could be provided. Finality in respect of the pricing of medicines needs to be achieved. This applies not only to the dispensing fee but also to the proposed benchmarking process and the proposed differentialbetween brand and generic medicines."

Pharmacoeconomics and "me-too" Drug Research and Development / 中国药房

OBJECTIVE:To discuss how can the pharmacoeconomics be applied in "me-too" drug research and development.METHODS:The current situation of "me-too" drugs,i.e.the advantageous and disadvantageous of "me-too" drug research and development was discussed.Then,the necessity and the application stage of pharmacoeconomics in "me-too" drug research and development,and issues regarding the option of pharmacoeconomic evaluation methods as well as the possible problems and the countermeasures in the application were addressed.RESULTS & CONCLUSION:Pharmacoeconomics can help promote the rationality of "me-too" drug research and development.

Proportions of Cells Expressing CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, or CD13,33+/CD34+ in the Regenerating Bone Marrows During Complete Remission of Acute Leukemia or After Bone Marrow Transplantation / 대한진단검사의학회지

BACKGROUND: The hemopoietic stem cells increase in number during the regeneration after chemotherapy or bone marrow transplantation (BMT). Although the proportion of hemopoietic stem cells and their differentiation have been studied by immunophenotyping using the flow cytometry, no substantial research efforts have been directed toward the regenerating marrow. We attempted to discover the proportions of undifferentiated stem cells, committed stem cells, B cell precursors, and myeloid precursors in the regenerating bone marrows during complete remission (CR) and after engraftment of BMT. METHODS: Bone marrow samples from 82 patients with acute leukemia in CR and from 25 patients after BMT engraftment, along with 22 control samples, were used to find the numbers of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells in the large lymphocyte gate by flow cytometry. We cross-analyzed our results in terms of groups: CR, BMT, and initial diagnosis groups. We performed significance tests on age, relapse, chromosomal abnormalities, clinical outcomes, and initial immunophenotypes of the leukemic cells. RESULTS: The proportions of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells are more highly distributed in acute B-lymphoblastic leukemia than the normal group and also in the CR than the BMT group. CD19+/CD34+ cells were increased in the relapse group and CD38+/ CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells were increased in the group with chromosomal abnormality. The results were irrelevant to the initial immunophenotype of the leukemic blasts. CONCLUSIONS: The increases of the markers spanned too widely to apply one specific cutoff value to analyze them. They seemed to be the results of normal regeneration, irrelevant to relapse or initial immunophenotype of leukemic blasts.

One standard of care for all is not always practical.

Multiple research guidelines address the issue of standard of care in international collaborative research. These guidelines fail to appreciate that differing standards may be present within the same country, which makes their application sometimes impracticable. In circumstances where ethics review committees follow one of these guidelines entirely and to the hilt, some relevant and useful research is rejected while the way for "me too" drug trials is paved. It should be acceptable to hold different researchers to separate standards of care on the basis of their intentions, their financial resources, their ultimate gains from the research, and subsequent utilisation of the results of the research, even when these researchers come from the same country where the research is being conducted.

Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis

Lactococcus lactis, the model lactic acid bacterium, is a good candidate for heterologous protein production in both foodstuffs and the digestive tract. We attempted to produce Streptomyces tendae antifungal protein 1 (Afp1) in L. lactis with the objective of constructing a strain able to limit fungal growth. Since Afp1 activity requires disulfide bond (DSB) formation and since intracellular redox conditions are reportedly unfavorable for DSB formation in prokaryotes, Afp1 was produced as a secreted form. An inducible expression-secretion system was used to drive Afp1 secretion by L. lactis; Afp1 was fused or not with LEISSTCDA, a synthetic propeptide (LEISS) that has been described to be a secretion enhancer. Production of Afp1 alone was not achieved, but production of LEISS-Afp1 was confirmed by Western blot and immunodetection with anti-Afp1 antibodies. This protein (molecular mass: 9.8 kDa) is the smallest non-bacteriocin heterologous protein ever reported to be secreted in L. lactis via the Sec-dependent pathway. However, no anti-fungal activity was detected, even in concentrated samples of induced supernatant. This could be due to a too low secretion yield of Afp1 in L. lactis, to the absence of DSB formation, or to an improper DSB formation involving the additional cysteine residue included in LEISS propeptide. This raises questions about size limits, conformation problems, and protein secretion yields in L. lactis.