Five-year microevolution of a multidrug-resistant Mycobacterium tuberculosis strain within a patient with inadequate compliance to treatment.

BACKGROUND: Whole-genome sequencing has shown that the Mycobacterium tuberculosis infection process can be more heterogeneous than previously thought. Compartmentalized infections, exogenous reinfections, and microevolution are manifestations of this clonal complexity. The analysis of the mechanisms causing the microevolution -the genetic variability of M. tuberculosis at short time scales- of a parental strain into clonal variants with a patient is a relevant issue that has not been yet completely addressed. To our knowledge, a whole genome sequence microevolution analysis in a single patient with inadequate adherence to treatment has not been previously reported. CASE PRESENTATION: In this work, we applied whole genome sequencing analysis for a more in-depth analysis of the microevolution of a parental Mycobacterium tuberculosis strain into clonal variants within a patient with poor treatment compliance in Argentina. We analyzed the whole-genome sequence of 8 consecutive Mycobacterium tuberculosis isolates obtained from a patient within 57-months of intermittent therapy. Nineteen mutations (9 short-term, 10 fixed variants) emerged, most of them associated with drug resistance. The first isolate was already resistant to isoniazid, rifampicin, and streptomycin, thereafter the strain developed resistance to fluoroquinolones and pyrazinamide. Surprisingly, isolates remained susceptible to the pro-drug ethionamide after acquiring a frameshift mutation in ethA, a gene required for its activation. We also found a novel variant, (T-54G), in the 5' untranslated region of whiB7 (T-54G), a region allegedly related to kanamycin resistance. Notably, discrepancies between canonical and phage-based susceptibility testing to kanamycin were previously found for the isolate harboring this mutation. In our patient, microevolution was mainly driven by drug selective pressure. Rare short-term mutations fixed together with resistance-conferring mutations during therapy. CONCLUSIONS: This report highlights the relevance of whole-genome sequencing analysis in the clinic for characterization of pre-XDR and MDR resistance profile, particularly in patients with incomplete and/or intermittent treatment.

Dosing of caspofungin based on a pharmacokinetic/pharmacodynamic index for the treatment of invasive fungal infections in critically ill patients on continuous venovenous haemodiafiltration.

INTRODUCTION: The study objective was to evaluate the efficacy of different dosages of caspofungin in the treatment of invasive candidiasis and aspergillosis, in relation to the probability of pharmacokinetic/pharmacodynamic (PK/PD) target attainment, using modelling and Monte Carlo simulations in critically ill adult patients on continuous haemodiafiltration. METHODS: Critically ill adult patients on continuous venovenous haemodiafiltration treated with caspofungin were analysed. A population PK model was developed. Four caspofungin dosing regimens were simulated: the licensed regimen, 70 mg/day, 100 mg/day or 200 mg/day. A PK/PD target was defined as the ratio between the area under the caspofungin concentration-time curve over 24 hours and the minimal inhibitory concentration (AUC/MIC) for candidiasis or the minimal effective concentrations (AUC/MEC) for Aspergillus spp. Target attainment based on preclinical target for Candida and Aspergillus was assessed for different MIC or MEC, respectively. RESULTS: Concentration-time data were described by a two-compartment model. Body-weight and protein concentration were the only covariates identified by the model. Goodness-of-fit plots and bootstrap analysis proved the model had a satisfactory performance. As expected, a higher maintenance dose resulted in a higher exposure. Target attainment was >90% for candidiasis (MIC≤0.06 mg/L) and aspergillosis (MEC≤0.5 mg/L), irrespective of the dosing regimen, but not for C. parapsilosis. Standard regimen was insufficient to reach the target for C. albicans and C. parapsilosis with MIC≥0.1 mg/L. CONCLUSION: The licensed regimen of caspofungin is insufficient to achieve the PK/PD targets in critically ill patients on haemodiafiltration. The determination of MICs will enable dose scheme selection.

Terapia Su Jok en pacientes con diagnóstico de hidrosalpinx / Su Jok therapy in patients with hydrosalpinx diagnosis

Se realizó un estudio cuasiexperimental de 56 pacientes con diagnóstico de hidrosalpinx, que acudieron a la consulta de Medicina Natural y Tradicional del Hospital General Docente Dr Juan Bruno Zayas Alfonso de Santiago de Cuba - remitidas de las consultas de Ginecología e Infertilidad -, desde septiembre de 2014 hasta diciembre de 2015, para evaluar la respuesta a la terapia Su Jok. Entre las variables analizadas figuraron: edad, factores de riesgo, manifestaciones clínicas, síndromes tradicionales, evolución clínica y respuesta al tratamiento. En la casuística predominaron las féminas de 30-39 años, la enfermedad inflamatoria pélvica y las infecciones de trasmisión sexual como antecedentes patológicos (71,4 por ciento), el dolor y la secreción vaginal como síntomas principales, así como el estancamiento de Qi de hígado, Qi de riñón no firme y frío-humedad en bazo como síndromes más frecuentes. Finalmente, se logró una evolución favorable y una respuesta adecuada al tratamiento(AU)

A quasi-experiment of 56 patients with hydrosalpinx diagnosis that went to the Natural and Traditional Medicine Service of Dr Juan Bruno Zayas Alfonso Teaching General Hospital in Santiago de Cuba - referred from the Gynecology and Infertility Services -, was carried out from September, 2014 to December, 2015, to evaluate the response to Su Jok therapy. Among the analyzed variables there were: age, risk factors, clinical features, traditional syndromes, clinical course and response to the treatment. In the case material there was a prevalence of women aged 30-39, pelvic inflammatory disease and sexually transmitted infections as pathological history (71.4 percent), pain and vaginal secretion as main symptoms, as well as Qi liver stagnation, not firm Qi kidney and coldness-humidity in spleen as most frequent syndromes. Finally, there was a favorable clinical course and an appropriate response to the treatment(AU)

Terapia Su Jok en pacientes con diagnóstico de hidrosalpinx / Su Jok therapy in patients with hydrosalpinx diagnosis

Se realizó un estudio cuasiexperimental de 56 pacientes con diagnóstico de hidrosalpinx, que acudieron a la consulta de Medicina Natural y Tradicional del Hospital General Docente Dr Juan Bruno Zayas Alfonso de Santiago de Cuba - remitidas de las consultas de Ginecología e Infertilidad -, desde septiembre de 2014 hasta diciembre de 2015, para evaluar la respuesta a la terapia Su Jok. Entre las variables analizadas figuraron: edad, factores de riesgo, manifestaciones clínicas, síndromes tradicionales, evolución clínica y respuesta al tratamiento. En la casuística predominaron las féminas de 30-39 años, la enfermedad inflamatoria pélvica y las infecciones de trasmisión sexual como antecedentes patológicos (71,4 por ciento), el dolor y la secreción vaginal como síntomas principales, así como el estancamiento de Qi de hígado, Qi de riñón no firme y frío-humedad en bazo como síndromes más frecuentes. Finalmente, se logró una evolución favorable y una respuesta adecuada al tratamiento

A quasi-experiment of 56 patients with hydrosalpinx diagnosis that went to the Natural and Traditional Medicine Service of Dr Juan Bruno Zayas Alfonso Teaching General Hospital in Santiago de Cuba - referred from the Gynecology and Infertility Services -, was carried out from September, 2014 to December, 2015, to evaluate the response to Su Jok therapy. Among the analyzed variables there were: age, risk factors, clinical features, traditional syndromes, clinical course and response to the treatment. In the case material there was a prevalence of women aged 30-39, pelvic inflammatory disease and sexually transmitted infections as pathological history (71.4 percent), pain and vaginal secretion as main symptoms, as well as Qi liver stagnation, not firm Qi kidney and coldness-humidity in spleen as most frequent syndromes. Finally, there was a favorable clinical course and an appropriate response to the treatment

[Not Available]. / Nutritional approach of inpatients with anorexia nervosa.

Anorexia nervosa (AN) is the most prevalent of eating disorders in children and adolescents, and its treatment is long and complex, involving a multidisciplinary team. Nutritional rehabilitation and restoration of a healthy body weight is one of the central goals in the initial stages of inpatient treatment. However, current recommendations on initial energy requirements for these patients are inconsistent, with a clear lack of controlled studies, available scientific evidence and global consensus on the most effective and safe refeeding practices in hospitalized adolescents with anorexia nervosa (AN). Conservative refeeding recommendations have been classically established in order to prevent the refeeding syndrome. Nevertheless, various works have recently appeared advocating a higher initial caloric intake, without observing more complications or refeeding syndrome, and allowing a shorter average stay. We present our experience in the treatment of restricting AN with a conservative progressive treatment. We have obtained good results with this approach, which was well tolerated by patients, with no observing complications. As a consequence, the medical team could establish a pact about the therapeutic goals with the patients in an easier way.

Nutritional approach of inpatients with anorexia nervosa / Abordaje nutricional de pacientes ingresados con anorexia nerviosa

Anorexia nervosa (AN) is the most prevalent of eating disorders in children and adolescents, and its treatment is long and complex, involving a multidisciplinary team. Nutritional rehabilitation and restoration of a healthy body weight is one of the central goals in the initial stages of inpatient treatment. However, current recommendations on initial energy requirements for these patients are inconsistent, with a clear lack of controlled studies, available scientific evidence and global consensus on the most effective and safe refeeding practices in hospitalized adolescents with anorexia nervosa (AN). Conservative refeeding recommendations have been classically established in order to prevent the refeeding syndrome. Nevertheless, various works have recently appeared advocating a higher initial caloric intake, without observing more complications or refeeding syndrome, and allowing a shorter average stay. We present our experience in the treatment of restricting AN with a conservative progressive treatment. We have obtained good results with this approach, which was well tolerated by patients, with no observing complications. As a consequence, the medical team could establish a pact about the therapeutic goals with the patients in an easier way (AU)

La anorexia nerviosa (AN) es el trastorno del comportamiento alimentario más prevalente en niños y adolescentes; su tratamiento es largo y complejo, e involucra a múltiples profesionales. La rehabilitación nutricional y la recuperación de un peso corporal normal es uno de los objetivos centrales en las fases iniciales del tratamiento del paciente ingresado. Sin embargo, las recomendaciones actuales sobre los requerimientos energéticos iniciales para estos pacientes son inconsistentes, con una clara ausencia de estudios controlados, evidencia científica disponible y consenso global sobre la forma de realimentación más efectiva y segura en adolescentes ingresados con anorexia nerviosa (AN). Clásicamente se han recomendado una realimentación conservadora para prevenir el síndrome de realimentación. No obstante, han aparecido recientemente varios trabajos recomendado una ingesta calórica inicial más elevada, sin observar más complicaciones ni síndrome de realimentación, y asociadas a estancias medias más cortas. Presentamos aquí nuestra experiencia en el tratamiento de la AN restrictiva con un tratamiento progresivo conservador. Hemos obtenido buenos resultados con este abordaje, bien tolerando por los pacientes, y sin observar complicaciones. Gracias a él, el equipo médico pudo establecer más fácilmente un acuerdo sobre los objetivos terapéuticos con el paciente (AU)

3-H-[1,2]Dithiole as a New Anti-Trypanosoma cruzi Chemotype: Biological and Mechanism of Action Studies.

The current pharmacological Chagas disease treatments, using Nifurtimox or Benznidazole, show limited therapeutic results and are associated with potential side effects, like mutagenicity. Using random screening we have identified new chemotypes that were able to inhibit relevant targets of the Trypanosoma cruzi. We found 3H-[1,2]dithioles with the ability to inhibit Trypanosoma cruzi triosephosphate isomerase (TcTIM). Herein, we studied the structural modifications of this chemotype to analyze the influence of volume, lipophilicity and electronic properties in the anti-T. cruzi activity. Their selectivity to parasites vs. mammalian cells was also examined. To get insights into a possible mechanism of action, the inhibition of the enzymatic activity of TcTIM and cruzipain, using the isolated enzymes, and the inhibition of membrane sterol biosynthesis and excreted metabolites, using the whole parasite, were achieved. We found that this structural framework is interesting for the generation of innovative drugs for the treatment of Chagas disease.

Production of biodiesel from vegetable oil and microalgae by fatty acid extraction and enzymatic esterification.

The aim of this work was to obtain biodiesel (methyl esters) from the saponifiable lipids (SLs) fraction of the microalga Nannochloropsis gaditana, whose biomass dry weight contains 12.1 wt% of these lipids. SLs were extracted from the microalga as free fatty acids (FFAs) for subsequent transformation to methyl esters (biodiesel) by enzymatic esterification. Extraction as FFAs rather than as SLs allows them to be obtained with higher purity. Microalgal FFAs were obtained by direct saponification of lipids in the biomass and subsequent extraction-purification with hexane. Esterification of FFAs with methanol was catalysed by lipase Novozym 435 from Candida antarctica. Stability studies of this lipase in the operational conditions showed that the esterification degree (ED) attained with the same batch of lipase remained constant over six reaction cycles (36 h total reaction time). The optimal conditions attained for 4 g of FFAs were 25°C, 200 rpm, methanol/FFA molar ratio of 1.5:1, Novozym 435/FFA ratio of 0.025:1 w/w and 4 h reaction time. In these conditions the ED attained was 92.6%, producing a biodiesel with 83 wt% purity from microalgal FFAs. Several experimental scales were tested (from 4 to 40 g FFAs), and in all cases similar EDs were obtained.

Burden of laboratory-confirmed Campylobacter infections in Guatemala 2008-2012: results from a facility-based surveillance system.

INTRODUCTION: Campylobacteriosis is one of the leading causes of gastroenteritis worldwide. This study describes the epidemiology of laboratory-confirmed Campylobacter diarrheal infections in two facility-based surveillance sites in Guatemala. METHODS: Clinical, epidemiologic, and laboratory data were collected on patients presenting with acute diarrhea from select healthcare facilities in the departments of Santa Rosa and Quetzaltenango, Guatemala, from January 2008 through August 2012. Stool specimens were cultured for Campylobacter and antimicrobial susceptibility testing was performed on a subset of isolates. Multidrug resistance (MDR) was defined as resistance to ≥3 antimicrobial classes. RESULTS: Campylobacter was isolated from 306 (6.0%) of 5137 stool specimens collected. For children <5 years of age, annual incidence was as high as 1288.8 per 100,000 children in Santa Rosa and 185.5 per 100,000 children in Quetzaltenango. Among 224 ambulatory care patients with Campylobacter, 169 (75.5%) received metronidazole or trimethoprim-sulfamethoxazole, and 152 (66.7%) received or were prescribed oral rehydration therapy. Antimicrobial susceptibilities were tested in 96 isolates; 57 (59.4%) were resistant to ciprofloxacin and 12 (12.5%) were MDR. CONCLUSION: Campylobacter was a major cause of diarrhea in children in two departments in Guatemala; antimicrobial resistance was high, and treatment regimens in the ambulatory setting which included metronidazole and trimethoprim-sulfamethoxazole and lacked oral rehydration were sub-optimal.

Urine melatonin and citrate excretion during the elite swimmers' training season.

The objectives of the study were to evaluate the influence of a whole training season on 6-sulfatoxymelatonin (αMT6s) and citrate excretion in 12 elite swimmers. Urine samples were obtained (before bedtime and after waking up) at the beginning of the season, basic training, macrocycle I, tapering and macrocycle II stages. For αMT6s, at basic training, mainly with aerobic training, the evening concentrations were significantly lower (P < 0.01; P < 0.05) than at the beginning, tapering and macrocycle II stages. At macrocycle II stage, with the maximal training workload, the total sum (evening plus morning) was significantly higher (P < 0.05) than at the beginning, basic training and macrocycle I stages. The ratio (morning/evening) that represents the capacity to produce melatonin at night depending on the evening values at the basic training stage and the nocturnal increment at the macrocycle II stage were significantly higher (P < 0.01) than at all other stages. Citrate morning values at basic training and tapering stages were significantly lower (P < 0.01) than in the evening indicating that a metabolic recuperation took place. The total sum significantly decreased (P < 0.05) as the aerobic training progressed from the beginning to macrocycle I. The basic training ratio (morning/evening) was significantly lower compared to the beginning and macrocycle II stages, and the nocturnal increment was significantly higher (P < 0.05) compared to the beginning. Melatonin and citrate represent complementary markers that could be used to evaluate the assimilation of the training workload by noninvasive methods.