Characteristics of a large population of patients with refractory epilepsy attending tertiary referral centers in Italy

P>The characteristics of 1,124 consecutive adults and children with refractory epilepsy attending 11 tertiary referral centers in Italy were investigated at enrollment into a prospective observational study. Among 933 adults (age 16-86 years), the most common syndromes were symptomatic (43.7%) and cryptogenic (39.0%) focal epilepsies, followed by idiopathic (8.1%) and cryptogenic/symptomatic generalized (6.2%) epilepsies. The most common syndrome among 191 children was symptomatic focal epilepsy (35.1%), followed by cryptogenic focal (18.8%), cryptogenic/symptomatic generalized (18.3%), undetermined whether focal or generalized (16.8%), and idiopathic generalized (7.3%). Primarily and secondarily generalized tonic-clonic seizures were reported in 27.8% of adults and 16.8% of children. The most commonly reported etiologies were mesial temporal sclerosis (8.0%) and disorders of cortical development (6.2%) in adults, and disorders of cortical development (14.7%) and nonprogressive encephalopathies (6.8%) in children. More than three-fourths of subjects in both age groups were on antiepileptic drug (AED) polytherapy.

Pediatric glioblastoma cell line shows different patterns of expression of transmembrane ABC transporters after in vitro exposure to vinblastine

Resistance to drug is a major cause of treatment failure in pediatric brain cancer. The multidrug resistance (MDR) phenotype can be mediated by the superfamily of adenosine triphosphate-binding cassette (ABC) transporters. The dynamics of expression of the MDR genes after exposure to chemotherapy, especially the comparison between pediatric brain tumors of different histology, is poorly described. To compare the expression profiles of the multidrug resistance genes ABCB1, ABCC1, and ABCG2 in different neuroepithelial pediatric brain tumor cell lines prior and following short-term culture with vinblastine. Immortalized lineages from pilocytic astrocytoma (R286), anaplasic astrocytoma (UW467), glioblastoma (SF188), and medulloblastoma (UW3) were exposed to vinblastine sulphate at different schedules (10 and 60 nM for 24 and 72 h). Relative amounts of mRNA expression were analyzed by real-time quantitative polymerase chain reaction. Protein expression was assessed by immunohistochemistry for ABCB1, ABCC1, and ABCG2. mRNA expression of ABCB1 increased together with augmenting concentration and time of exposure to vinblastine for R286, UW467, and UW3 cell lines. Interestingly, ABCB1 levels of expression diminished in SF188. Following chemotherapy, mRNA expression of ABCC1 decreased in all cell lines other than glioblastoma. ABCG2 expression was influenced by vinblastine only for UW3. The mRNA levels showed consistent association to protein expression in the selected sets of cell lines analyzed. The pediatric glioblastoma cell line SF188 shows different pattern of expression of multidrug resistance genes when exposed to vinblastine. These preliminary findings may be useful in determining novel strategies of treatment for neuroepithelial pediatric brain tumors.

Addressing overtreatment in patients with refractory epilepsy at a tertiary referral centre in Brazil

Background. Patients with refractory epilepsy often have impaired quality of life (QOL) as a consequence of seizures and adverse effects of antiepileptic drugs. We assessed the impact of adverse effects on QOL and the utility of a structured instrument to help the physician manage adverse effects in patients with refractory epilepsy. Methods. Clinical characteristics, drug treatment and adverse effects were evaluated in 102 patients with refractory epilepsy at a single tertiary referral centre. The Adverse Events Profile (AEP) and Quality of Life in Epilepsy-31 (QOLIE-31) questionnaires were completed at baseline and after six months. At baseline, patients with a high burden of adverse effects (AEP scores >= 45) were randomized to an intervention or control group. AEP scores in the intervention group were available to the physician as an instrument to help to reduce adverse effects. Results. Ninety-five patients (93.1%) were on polytherapy. Sixty-six completed the questionnaires and, of these, 43 (65.1%) had a high AE burden and were randomized to the intervention and control group. QOLIE-31 scores were inversely correlated with AEP scores at both visits. Among randomized patients, AEP scores tended to decrease between the baseline and the final visit without significant differences between groups (intervention group: 54.1 +/- 6.1 vs 51.1 +/- 9.1; control group: 55.8 +/- 5.8 vs 50.5 +/- 12.2). QOLIE-31 scores did not change substantially between visits (intervention group: 45.9 +/- 17.4 vs 48.4 +/- 14; control group: 47.5 +/- 15.7 vs 45.2 +/- 18.9). Conclusion. A significant proportion of patients had a high toxicity burden which had an impact on their QOL. Reduction of over-treatment is a difficult challenge which cannot be addressed solely by providing a structured assessment of adverse effects, but requires a more comprehensive approach aimed at optimizing the many components of the management strategy.

Efficacy of praziquantel against Schistosoma japonicum: field evaluation in an area with repeated chemotherapy compared with a newly identified endemic focus in Hunan, China

This study conducted in 1999/2000 was designed to evaluate the efficacy of praziquantel against Schistosoma japonicum in an area with repeated chemotherapy (Area A) compared with a newly identified endemic focus (Area B) in Hunan Province, China. The population size was 2015 and 2180 in Areas A and B, respectively, of which 1129 and 1298 subjects received stool examination. A total of 230 subjects were identified by the Kato-Katz technique (4 smears per person) as being infected with S. japonicum, 124 in Area A (prevalence 11 %) and 106 in Area B (prevalence 8.2%). They were treated with a single oral dose of praziquantel (40 mg/kg) in the non-transmission season. A follow-up stool examination was made 50 days after treatment. Among the 220 cases followed, 22 were found stool-egg-positive, with an overall cure rate of 90 %, and 99 % reduction of infection intensity (eggs per gram stool). No significant difference was found in cure rates between the 2 areas (89.7% vs 90.3%). The efficacy of the drug in the area with repeated chemotherapy was not significantly different from that in the newly identified endemic focus. This study, therefore, suggests that the efficacy of praziquantel against S. japonicum has not changed in the Dongting Lake region after more than 14 years of mass chemotherapy, and there is no evidence of tolerance or resistance of S. japonicum against praziquantel.

Mutation rates in the dihydrofolate reductase gene of Plasmodium falciparum

A new method has been established to define the limits on a spontaneous mutation rate for a gene in Plasmodium falciparum. The method combines mathematical modelling and large-scale in vitro culturing and calculates the difference in mutant frequencies at 2 separate time-points. We measured the mutation rate at 2 positions in the dihydrofolate reductase (DHFR) gene of 3D7, a pyrimethamine-sensitive line of P. fulciparum. This line was re-cloned and an effectively large population was treated with a selective pyrimethamine concentration of 40 nM. We detected point mutations at codon-46 (TTA to TCA) and codon-108 (ACC to AAC), resulting in serine replacing leucine and asparagine replacing serine respectively in the corresponding gene product. The substitutions caused a decrease in pyrimethamine sensitivity. By mathematical modelling we determined that the mutation rate at a given position in DHFR was low and occurred at less than 2(.)5 x 10(-9) mutations/DHFR gene/replication. This result has important implications for Plasmodium genetic diversity and antimalarial drug therapy by demonstrating that even with lon mutation rates anti-malarial resistance will inevitably arise when mutant alleles are selected under drug pressure.

Gag-Pol supplied in trans is efficiently packaged and supports viral function in human immunodeficiency virus type 1

The intracellular trafficking and subsequent incorporation of Gag-Pol into human immunodeficiency virus type 1 (HIV-1) remains poorly defined. Gag-Pol is encoded by the same mRNA as Gag and is generated by ribosomal frameshifting. The multimerization of Gag and Gag-Pol is an essential step in the formation of infectious viral particles. In this study, we examined whether the interaction between Gag and Gag-Pol is initiated during protein translation in order to facilitate the trafficking and subsequent packaging of Gag-Pol into the virion. A conditional cotransfection system was developed in which virion formation required the coexpression of two HIV-1-based plasmids, one that produces both Gag and Gag-Pol and one that only produces Gag-Pol. The Gag-Pol proteins were either immunotagged with a His epitope or functionally tagged with a mutation (K65R) in reverse transcriptase that is associated with drug resistance. Gag-Pol packaging was assessed to determine whether the Gag-Pol incorporated into the virion was preferentially packaged from the plasmid that expressed both Gag and Gag-Pol or whether it could be packaged from either plasmid. Our data show that translation of Gag and Gag-Pol from the same mRNA is not critical for virion packaging of the Gag-Pol polyprotein or for viral function.

Desenvolvimento e padronização de um método para determinação da sensibilidade do mycobacterium tuberculosis a fármacos contra tuberculose

A resistência a fármacos antituberculose tem constituído uma grande ameaça ao controle da tuberculose em âmbito mundial. A sua detecção precoce permite ao médico instituir um esquema de tratamento mais adequado ao paciente e consequentemente quebrar a cadeia de transmissão dos bacilos. Os testes de sensibilidade a antimicrobianos atuais, embora eficientes, são caros e/ou demorados e/ou trabalhosos. Com base nesta premissa, nos propusemos a desenvolver e padronizar um método fenotípico direto para determinação da sensibilidade do Mycobacterium tuberculosis a antimicrobianos de primeira linha do tratamento da tuberculose. Para o desenvolvimento deste novo teste, utilizaram-se os princípios do método das proporções e do exame de cultura pelo método de Ogawa–Kudoh. O estudo foi dividido em duas fases. A primeira, caracterizada pelo desenvolvimento e padronização do método proposto e a segunda, pela análise da concordância entre o método desenvolvido e o método do MGIT (padrão-ouro). Na primeira fase, foram realizados diversos ensaios para definir: os volumes de absorção e de liberação de líquidos de diferentes tipos de swab, o meio de cultura, as concentrações dos antimicrobianos e o tempo de leitura/interpretação das culturas. Além disso, foi verificado se a amostra deveria ou não ser diluída. Com base nos resultados destes ensaios, padronizou-se o método com: swab comercial, em meio de cultura Ogawa- Kudoh contendo separadamente 0,2 μg/mL de isoniazida, 40,0 μg/mL de rifampicina, 10,0 μg/mL de estreptomicina e 500,0 μg/mL de ácido para-nitrobenzóico. Padronizou-se ainda a inoculação da amostra de escarro de forma direta, ou seja, sem diluir e a leitura/interpretação do resultado do teste no período entre 21 e 28 dias. A análise comparativa entre este método e o teste de sensibilidade a antimicrobianos no sistema MGIT realizada na segunda fase do projeto indicou um índice kappa igual a 1,000, ou seja, uma concordância muito boa em relação ao padrão-ouro. Diante desses resultados promissores, acreditamos que o método desenvolvido apresente um grande potencial para ser utilizado em laboratórios com pouca infra-estrutura, por ser de baixo custo, fácil execução e relativamente rápido.

Caracterização molecular, formação de biofilme e susceptibilidade a antimicrobianos de isolados de E. coli de aderência difusa Afa/Dr e não Afa/Dr

A Escherichia coli de aderência difusa (DAEC), um patotipo diarreiogênico de E. coli, corresponde a um grupo heterogêneo sem marcador de virulência comum a todos os isolados e de papel controverso na diarreia infantil. O objetivo deste estudo foi caracterizar genotipica e fenotipicamente amostras de DAEC, portadoras e não portadoras de adesinas Afa/Dr, isoladas de crianças com e sem diarreia. Em 70 amostras de DAEC, PCR foi realizado para pesquisa de genes descritos em DAEC, EAEC ou UPEC, que codificam: (i) oito adesinas fimbriais e afimbriais (fimH, papC, sfa, aggA, aafA, agg3A, aidA/aah, afaC); (ii) cinco toxinas (pet, astA, set1A, sat, hlyA); (iii) três proteínas captadoras/receptora de ferro (irp2, iucA, chuA/shuA); (iv) invasina (daaD) e; antígeno 43 (agn43). Ensaio de formação de biofilme foi realizado a partir da bactéria cultivada em caldo Luria-Bertani e inoculada em placas de poliestireno com DMEM suplementado com 0,4% glicose. A leitura da densidade ótica (DO490) foi realizada após coloração com safranina. Soroaglutinação para 23 antígenos O (Probac do Brasil) foi realizada em 50% das DAEC. Método de difusão de disco foi realizado para testar a suscetibilidade a 13 antimicrobianos. A presença de pelo menos um gene que codifica adesinas, toxinas, proteínas captadoras/receptora de ferro, invasina ou antígeno 43 foram encontrados em 58,6%, 51,4%, 80%, 48,6% e 57,1%, respectivamente, com os genes fimH, irp2, agn43, iucA, chuA/shuA, presentes em mais de 50% das amostras. Gene afaC+ (PCR) e/ou sonda afaBC+ (hibridização de colônias) classificou 50% das DAEC como Afa/Dr, sendo pet, sat, irp2, iucA, chuA/shuA e agn43 significantes nessas amostras (p<0,05). Do total das DAEC, 44,3% foram formadoras de biofilme, igualmente distribuídas entre as Afa/Dr e não Afa/Dr, e nenhum gene foi associado com esse fenótipo. Sorologia de 35 amostras evidenciou os seguintes sorogrupos: 1 O29, 2 O125, 2 O127 e 7 O86. Todas as O86 foram de DAEC Afa/Dr. Maiores frequências de resistência antimicrobiana foram encontradas para ampicilina (55,7%), sulfametoxazol/trimetoprim (35,7%) e tetraciclina (28,6%) e o perfil resistente/intermediário para amoxicilina/ácido clavulânico, ampicilina, sulfametoxazol/trimetoprim foi significante nas DAEC Afa/Dr, assim como a multi-droga resistência (p<0,05). Em conclusão, observou-se: (i) alta frequência de fimH e pet e presença de agn43, até então não descrito em DAEC, em frequências similares àquelas encontradas em EAEC, UPEC e EAEC/UPEC, respectivamente; (ii) que as amostras de DAEC Afa/Dr e não Afa/Dr constituíram grupos com perfis genéticos diferenciados entre si; (iii) poucos sorogrupos foram encontrados entre as DAEC; (iv) frequências de resistência menores quando comparado com as poucas descrições em DAEC, sugerindo uma menor pressão seletiva da população do presente estudo e; (v) amostras de DAEC Afa/Dr podem representar um importante reservatório de genes de resistência a antimicrobianos, além de diversos fatores de virulência.

Differential expression of CDC25 phosphatases splice variants in human breast cancer cells

Background: CDC25 phosphatases control cell cycle progression by activating cyclin dependent kinases. The three CDC25 isoforms encoding genes are submitted to alternative splicing events which generate at least two variants for CDC25A and five for both CDC25B and CDC25C. An over-expression of CDC25 was reported in several types of cancer, including breast cancer, and is often associated with a poor prognosis. Nevertheless, most of the previous studies did not address the expression of CDC25 splice variants. Here, we evaluated CDC25 spliced transcripts expression in anti-cancerous drug-sensitive and resistant breast cancer cell lines in order to identify potential breast cancer biomarkers. Methods: CDC25 splice variants mRNA levels were evaluated by semi-quantitative RT-PCR and by an original real-time RT-PCR assay. Results: CDC25 spliced transcripts are differentially expres-sed in the breast cancer cell lines studied. An up-regulation of CDC25A2 variant and an increase of the CDC25C5/C1 ratio are associated to the multidrug-resistance in VCREMS and DOXOR breast cancer cells, compared to their sensitive counterpart cell line MCF-7. Additionally, CDC25B2 tran-script is exclusively over-expressed in VCREMS resistant cells and could therefore be involved in the development of certain type of drug resistance. Conclusions: CDC25 splice variants could represent interesting potential breast cancer prognostic biomarkers.

Impact of a training course on the quality of malaria diagnosis by microscopy in Angola

Background: In Angola, malaria is an endemic disease having a major impact on the economy. The WHO recommends testing for all suspected malaria cases, to avoid the presumptive treatment of this disease. In malaria endemic regions laboratory technicians must be very comfortable with microscopy, the golden standard for malaria diagnosis, to avoid the incorrect diagnosis. The improper use of medication promotes drug resistance and undesirable side effects. The present study aims to assess the impact of a three-day refresher course on the knowledge of technicians, quality of blood smears preparation and accuracy of microscopy malaria diagnosis, using qPCR as reference method. Methods: This study was implemented in laboratories from three hospitals in different provinces of Angola: Bengo, Benguela and Luanda. In each laboratory samples were collected before and after the training course (slide with thin and thick blood smears, a dried blood spot and a form). The impact of the intervention was evaluated through a written test, the quality of slide preparation and the performance of microscopy. Results: It was found a significant increase on the written test median score, from 52.5% to 65.0%. A total of 973 slides were analysed to evaluate the quality of thick and thin blood smears. Considering all laboratories there was a significant increase in quality of thick and thin blood smears. To determine the performance of microscopy using qPCR as the reference method we used 1,028 samples. Benguela presented the highest values for specificity, 92.9% and 98.8% pre and post-course, respectively and for sensitivity the best pre-course was Benguela (75.9%) and post-course Luanda (75.0%). However, no significant increase in sensitivity and specificity after the training course was registered in any laboratory analysed. Discussion: The findings of this study support the need of continuous refresher training for microscopists and other laboratory staff. The laboratories should have a quality control programme to supervise the diagnosis and also to assess the periodicity of new training. However, other variables needed to be considered to have a correct malaria diagnosis, such as adequate equipment and reagents for staining and visualization, good working conditions, motivated and qualified personnel.

Development of a high-throughput monitoring technique of bacteria photodynamic inactivation

Summary form only given. Bacterial infections and the fight against them have been one of the major concerns of mankind since the dawn of time. During the `golden years' of antibiotic discovery, during the 1940-90s, it was thought that the war against infectious diseases had been won. However currently, due to the drug resistance increase, associated with the inefficiency of discovering new antibiotic classes, infectious diseases are again a major public health concern. A potential alternative to antibiotic treatments may be the antimicrobial photodynamic inactivation (PDI) therapy. To date no indication of antimicrobial PDI resistance development has been reported. However the PDI protocol depends on the bacteria species [1], and in some cases on the bacteria strains, for instance Staphylococcus aureus [2]. Therefore the development of PDI monitoring techniques for diverse bacteria strains is critical in pursuing further understanding of such promising alternative therapy. The present works aims to evaluate Fourier-Transformed-Infra-Red (FT-IR) spectroscopy to monitor the PDI of two model bacteria, a gram-negative (Escherichia coli) and a gram-positive (S. aureus) bacteria. For that a high-throughput FTIR spectroscopic method was implemented as generally described in Scholz et al. [3], using short incubation periods and microliter quantities of the incubation mixture containing the bacteria and the PDI-drug model the known bactericidal tetracationic porphyrin 5,10,15,20-tetrakis (4-N, N, Ntrimethylammoniumphenyl)-porphyrin p-tosylate (TTAP4+). In both bacteria models it was possible to detect, by FTIR-spectroscopy, the drugs effect on the cellular composition either directly on the spectra or on score plots of principal component analysis. Furthermore the technique enabled to infer the effect of PDI on the major cellular biomolecules and metabolic status, for example the turn-over metabolism. In summary bacteria PDI was monitored in an economic, rapid (in minutes- , high-throughput (using microplates with 96 wells) and highly sensitive mode resourcing to FTIR spectroscopy, which could serve has a technological basis for the evaluation of antimicrobial PDI therapies efficiency.

Predictive factors for response to Lamivudine in chronic hepatitis B

BACKGROUND: Lamivudine has been shown to be an efficient drug for chronic hepatitis B (CHB) treatment. AIM: To investigate predictive factors of response, using a quantitative method with high sensitivity. METHODS: We carried out a prospective trial of lamivudine in 35 patients with CHB and evidence for viral replication, regardless to their HBeAg status. Lamivudine was given for 12 months at 300 mg daily and 150 mg thereafter. Response was considered when DNA was undetectable by PCR after 6 months of treatment. Viral replication was monitored by end-point dilution PCR. Mutation associated with resistance to lamivudine was detected by DNA sequencing in non-responder patients. RESULTS: Response was observed in 23/35 patients (65.7%) but only in 5/15 (33.3%) HBeAg positive patients. Only three pre-treatment variables were associated to low response: HBeAg (p = 0.006), high viral load (DNA-VHB > 3 x 10(6) copies/ml) (p = 0.004) and liver HBcAg (p = 0.0028). YMDD mutations were detected in 7/11 non-responder patients. CONCLUSIONS: HBeAg positive patients with high viral load show a high risk for developing drug resistance. On the other hand, HBeAg negative patients show a good response to lamivudine even with high viremia.

Role of ß-lactamase operon on mecA expression in Staphylococcus aureus

RESUMO: Os Staphylococcus aureus resistentes à meticilina (MRSA, do inglês “methicillin-resistant Staphylococcus aureus”) são um dos principais agentes responsáveis por infeções hospitalares. Os MRSA são resistentes a praticamente todos os antibióticos β-lactâmicos devido a dois mecanismos principais: produção de β-lactamase (bla), codificada pelo gene blaZ, e produção de uma proteína de ligação à penicilina (PBP2a, do inglês “penicillin binding protein 2”), codificada pelo gene mecA. Estes dois genes são regulados por sistemas homólogos, constituídos por um sensor-transdutor (BlaR1 e MecR1) e um repressor (BlaI e MecI), de tal modo que ambos os sistemas são capazes de co-regular os genes mecA e blaZ, embora com eficiências de indução muito diferentes. De facto, a indução mediada pelo sistema mecI-mecR1 é tão lenta que se acredita que este sistema não está funcional na maioria das estirpes MRSA. No entanto, dados recentes do nosso laboratório, demonstram a ausência de relação entre a presença do gene mecI e o nível de resistência à meticilina em estirpes MRSA epidémicas, e também que, o fenótipo de resistência da grande maioria das estirpes não é perturbado pela sobre-expressão em trans do repressor mecI. Curiosamente, as duas estirpes em que a expressão da resistência foi afetada pela sobre-expressão do mecI são negativas para o locus da β-lactamase, o que sugere que este locus pode interferir diretamente com a repressão do gene mecA mediada pelo MecI. Nesta tese de mestrado esta hipótese foi explorada usando estratégias de biologia molecular e ensaios fenotípicos da resistência aos -lactâmicos. Os resultados obtidos demonstram que a presença do plasmídeo nativo da β-lactamase não só anula a repressão mediada pelo MecI, como também aumenta o nível de resistência das estirpes parentais. Várias hipóteses foram então formuladas para explicar estas observações. Dados preliminares, em conjunto com evidências experimentais publicadas, sugerem que o BlaI forma hetero-dímeros com o MecI que, após a indução, são inativados eficientemente pelo BlaR1. Em conclusão, estes resultados apresentam novas perspetivas para o mecanismo de regulação do mecA e para uma nova importante função do operão da β-lactamase para o fenótipo das estirpes MRSA.-------------------ABSTRACT: Methicillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial pathogen and is also emerging in the community. MRSA is cross-resistant to virtually all β-lactam antibiotics and has acquired two main resistance mechanisms: production of β-lactamase (bla), coded by blaZ, and production of penicillin binding protein 2a (PBP2a), coded by mecA. Both genes are regulated by homologous sensor-transducers (BlaR1 and MecR1) and repressors (BlaI and MecI), and coregulation of mecA and blaZ by both systems has been demonstrated, although with remarkable different efficiencies. In fact, induction of mecA by mecI-mecR1 is so slow that it is believed it is not functional in most MRSA strains. However, recent data from our laboratory has unexpectedly demonstrated that not only there is no correlation between the presence of mecI gene and the resistance level in epidemic MRSA strains, but also that for most strains there were no significant changes on the resistance phenotype upon the mecI overexpression in trans. Interestingly, the two strains in which mecI overexpression affected the resistance expression were negative for the bla locus, suggesting that this locus may interfere directly with the MecI-mediated repression of mecA and account for those puzzling observations. In this master thesis we have explored this hypothesis using molecular biology strategies and phenotypic analysis of -lactam resistance. The data obtained demonstrate that the presence of a wild-type plasmid containing the bla locus not only disrupts the MecImediated repression, but also significantly enhances the expression of resistance. Several preliminary hypotheses were formulated to explain these observations and preliminary data, together with published evidence, support the working model that BlaI forms functional hetero-dimers with MecI, which upon induction are readily inactivated by BlaR1. These results provide new insights into the regulatory mechanism(s) of mecA and open new perspectives for the role of β-lactamase operon in the MRSA phenotype.

Analysis of HIV- type 1 protease and reverse transcriptase in Brazilian children failing highly active antiretroviral therapy (HAART)

The aim of this study was to evaluate the genotypic resistance profiles of HIV-1 in children failing highly active antiretroviral therapy (HAART). Forty-one children (median age = 67 months) receiving HAART were submitted to genotypic testing when virological failure was detected. cDNA was extracted from PBMCs and amplified by nested PCR for the reverse transcriptase and protease regions of the pol gene. Drug resistance genotypes were determined from DNA sequencing. According to the genotypic analysis, 12/36 (33.3%) and 6/36 (16.6%) children showed resistance and possible resistance, respectively, to ZDV; 5/36 (14%) and 4/36 (11.1%), respectively, showed resistance and possible resistance to ddI; 4/36 (11.1%) showed resistance to 3TC and D4T; and 3/36 (8.3%) showed resistance to Abacavir. A high percentage (54%) of children exhibited mutations conferring resistance to NNRTI class drugs. Respective rates of resistance and possible resistance to PIs were: RTV (12.2%, 7.3%); APV (2.4%, 12.1%); SQV(0%, 12.1%); IDV (14.6%, 4.9%), NFV (22%, 4.9%), LPV/RTV (2.4%, 12.1%). Overall, 37/41 (90%) children exhibited virus with mutations related to drug resistance, while 9% exhibited resistance to all three antiretroviral drug classes.

Evaluation of the in vitro activity of six antimicrobial agents against Neisseria gonorrhoeae

Use of antimicrobials for the treatment of gonorrhea started in 1930 with the utilization of sulfonamides. With the years other drugs were used for its treatment such as penicillin, tetracycline, spectinomycin, and others. Although highly specific in the beginning, these drugs, with time did not show anymore the expected therapeutic results because of aspects of chromosomal and plasmid-mediated resistance. The purpose of this study was to evaluate the susceptibility of Neisseria gonorrhoeae strains to six drugs used for its treatment (penicillin, tetracycline, cefoxitin, thiamphenicol, spectinomycin and ofloxacin) by the determination of minimal inhibitory concentrations of these drugs. We concluded that drugs, such as cefoxitin, thiamphenicol and spectinomycin still are excellent pharmacological agents for the treatment of gonorrhea. Penicillin, although still efficient, needs more attention regarding its use, as well as ofloxacin, because of the emergence of resistant strains. Tetracycline and its derivatives should be strongly contraindicated for the treatment of gonorrhea.