Evaluación del grado de adherencia al tratamiento intravenoso no antineoplásico de pacientes ambulatorios / Evaluation of the degree of adherence to non-antineoplastic intravenous treatment of ambulatory patients

Introducción: La falta de adherencia al tratamiento farmacológico es un problema prevalente y relevante en la evolución clínica.Objetivo: Evaluar el grado de adherencia al tratamiento intravenoso no antineoplásico de pacientes crónicos que acuden al hospital de día (HD), identificar los factores relacionados y analizar los desvíos de programación de la cita previa en el HD y su impacto en el Servicio de Farmacia (SF).Material y métodos: Estudio descriptivo longitudinal retrospectivo de tres años de duración (2017-2019) que incluyó a los pacientes que acudieron al HD a administrarse el tratamiento. Los datos de adherencia se extrajeron de los registros del SF y los datos demográficos-clínicos, de las historias clínicas electrónicas. El grado de adherencia se expresó en porcentaje (adherencia adecuada ≥90%). La asociación entre las variables y el grado de adherencia se estudió mediante test estadísticos de contraste de hipótesis.Resultados: Se incluyeron 300 pacientes, 60% mujeres, edad media 47 años. El 18% presentaron una adherencia inadecuada. La adherencia se asoció con la edad, el medicamento administrado, el intervalo posológico y la persistencia al tratamiento (p<0,05). El 5,6% del trabajo de HD se desvió de la programación suponiendo un trabajo adicional para el SF.Conclusiones: El grado de adherencia al tratamiento era inadecuado en casi una cuarta parte de la población. La edad, el medicamento infundido, el intervalo posológico y la persistencia al tratamiento se asociaron con la adherencia de los pacientes. Los desvíos en la programación de HD se tradujeron en trabajo sobreañadido para el SF. (AU)

Introduction: The lack of adherence to pharmacological treatment of patients with chronic diseases it is a relevant problem.Objective: To assess the degree of adherence to the non-chemotherapy intravenous treatment of chronic patients who came to the outpatient clinic (OC), to identify the possible specific factors related to therapeutic compliance and to analyze the appointment changes in the OC and its impact on the Pharmacy Department (PD).Material and methods: Retrospective longitudinal descriptive study of three years duration (2017-2019). This included patients who went to the OC to receive the treatment. Adherence data were extracted from the PD records and demographic-clinical data from the review of electronic health records. Besides, the degree of adherence was expressed as a percentage (adherence adequate ≥90%). The association between the variables studied and the degree of adherence was estimated by means of statistical tests of hypothesis contrast.Results: A total sample size of 300 patients were included, mean age 47 years, 60% women. Adherence of the treatment was inadequate in 18% of patients. The variables that showed a statistically significant association with adherence were the age, the drug delivered, the dosage interval and the persistence of treatment (p<0.05). 5.6% of OC work deviated from schedule and it meant additional work to the PD.Conclusions: The degree of adherence to the intravenous ambulatory treatment was inadequate in approximately a quarter of the population. The age, the infused drug, the dosage interval and the persistence of treatment were the variables that showed association with the adherence. Changes to OC programming resulted in over-added work for the PD. (AU)

More Than 25 Years Together: Basis for a Long-Lifespan Kidney Transplant.

Patients with graft survival for 20 years or more are not uncommon; they are called ultralong kidney recipients. It is interesting to know if there are patterns in donors and recipients that could be reproduced. A retrospective cohort with 22 adult patients with a kidney renal transplant performed more than 25 years ago is analyzed. The mean of age of the donors was 24 years (median, 21 years); 82% were men and the cause of death was mainly acute traumatic brain injury. Recipients had a mean age of 34 years (median, 36 years) at the time of transplant; the most common underlying renal disease was glomerular, without evidence of recurrence. A total of 16 patients had compatibility in HLA II (1 in 11 cases; 2 in 5 cases). Only 6 patients have had any episode of acute rejection; 3 of them have developed antibodies class I, but no donor-specific antibodies. In this retrospective cohort, increases in donor age are associated with poor renal function. The mean creatinine is 1.43 mg/dL (range, 0.97-2.14 mg/dL) and mean proteinuria is 653.43 mg/g (range, 55-3722 mg/g). The characteristics common in ultralong kidney recipients are young male donors, a shortage of episodes of rejection, and good HLA compatibility, especially in class II antigens.

The effect of body composition and serum inflammatory markers on the functional muscle-bone unit in premenopausal women.

BACKGROUND/OBJECTIVES: A number of recent studies dealing with the relationship between the effects of high body mass (BM) and fat mass (FM) on bone mass and strength exhibit a range of contrasting variations in their findings. These diverse findings have led to an ongoing controversy as to whether high BM and FM positively or negatively affect bone mass and strength. Excessive FM and the associated low-grade inflammation might overturn the higher mechanical stimulus arising from a higher BM. Therefore, we aimed at quantifying the functional muscle-bone unit in premenopausal women with markedly diverging body composition. SUBJECTS/METHODS: Sixty-four young women with BMs ranging from 50 to 113 kg and body fat percentages between 20.7% and 51.8% underwent jumping mechanography and peripheral quantitative computed tomography measurements. Maximum voluntary ground reaction force during multiple one-legged hopping (Fm1LH), as well as bone characteristics at 4, 14 and 38% of tibia length, were determined. Body composition was assessed by dual-energy X-ray absorptiometry, and serum inflammatory markers were analyzed from blood samples. RESULTS: Fm1LH predicted volumetric bone mineral content at the 14% site by 48.7%. Women with high body fat percentage had significantly higher Fm1LH, significantly lower relative bone mass, relative bone strength and relative bone area, as well as higher serum inflammatory markers in comparison to women with lower body fat percentage. CONCLUSIONS: In conclusion, high body fat percentage was associated with lower relative bone mass and strength despite normal habitual muscle force in premenopausal women, indicating that high body fat percentage compromised the functional muscle-bone unit in these individuals.

Causes for an Incomplete Donation Process at a Tertiary Care Hospital.

INTRODUCTION: This study aims to identify the causes for the incomplete donation process at a tertiary care hospital. MATERIALS AND METHODS: A descriptive, retrospective study was performed; all potential donors reported to the Transplant Service within the period of 2005 to 2014 were included. Descriptive statistics were used across frequencies and proportions for categorical variables, central tendency, and dispersion for continuous variables. RESULTS: The total number of deaths reported at the University Hospital (HU) was 8472, of which 815 (n = 815) were reported to COETRA ("Consejo Estatal de Trasplantes"). Among organ or tissue donors, the main known cause of death was head trauma (HT) in 26% (72). Cardiac arrest (CA) as cause of death provided the largest number of donations (141, 57%); of these, 102 (41%) were male and 39 (16%) were female. In comparison, brain death (BD) provided 104 (43%); of these, 65 (27%) were male, and 39 (16%) were female. The age interval was with a higher donation rate was 45 to 49 y (BD 18, CA 22). Donation request was not performed in 359 patients because of medical contraindication 60% (215), rapid deterioration 18% (64), and incomplete donation process 8% (27). Of 452 organ requests, 207 were not accomplished, because of body integrity 28% (57), family disagreement 20% (42), and no acceptance of BE 13% (26). CONCLUSIONS: Opportunity areas: (1) Ensure the notification of all deaths to Transplant Department for identification of potential donors; (2) Reduce rapid deterioration and raise number of completed donation protocols; (3) Increase the donation rate.

New point mutation in Golga3 causes multiple defects in spermatogenesis.

Mice with repro27 exhibit fully penetrant male-specific infertility associated with a nonsense mutation in the golgin subfamily A member 3 gene (Golga3). GOLGA3 is a Golgi complex-associated protein implicated in protein trafficking, apoptosis, positioning of the Golgi and spermatogenesis. In repro27 mutant mice, a point mutation in exon 18 of the Golga3 gene that inserts a pre-mature termination codon leads to an absence of GOLGA3 protein expression. GOLGA3 protein was undetectable in the brain, heart and liver in both mutant and control mice. Although spermatogenesis in Golga3(repro27) mutant mice appears to initiate normally, development is disrupted in late meiosis during the first wave of spermatogenesis, leading to significant germ cell loss between 15 and 18 days post-partum (dpp). Terminal Deoxynucleotidyl Transferase dUTP-mediated Nick End Labeling analysis showed elevated DNA fragmentation in meiotic germ cells by 12 dpp, suggesting apoptosis as a mechanism of germ cell loss. The few surviving post-meiotic round spermatids exhibited abnormal spermiogenesis with defects in acrosome formation, head and tail development and extensive vacuolization in the seminiferous epithelium. Analysis of epididymal spermatozoa showed significantly low sperm concentration and motility and in vitro fertilization with mutant spermatozoa was unsuccessful. Golga3(repro27) mice lack GOLGA3 protein and thus provide an in vivo tool to aid in deciphering the role of GOLGA3 in Golgi complex positioning, cargo trafficking and apoptosis signalling in male germ cells.

Atención farmacéutica en pacientes ingresados con insuficiencia renal / Pharmaceutical care in renal insufficiency inpatients

Objetivos: Valorar los resultados de un programa de ajuste posológico en pacientes hospitalizados con enfermedad renal, describir los medicamentos mayoritariamente implicados y determinar el grado de aceptación de la intervención farmacéutica realizada. Método: Estudio prospectivo, de intervención no aleatorizada, de 15 meses de duración en pacientes hospitalizados con función renal alterada (Creatinina sérica > 1,4 mg/dL) y en tratamiento con medicamentos que precisan ajuste en insuficiencia renal. La variable principal fue el porcentaje de adecuación posológica según la tasa de filtrado glomerular. También se evaluó la aceptación global por servicio clínico, el grupo farmacoterapéutico más prescrito de forma inadecuada, las monitorizaciones farmacocinéticas que se derivaron de la intervención y el carácter educativo de la misma. Resultados: Se identificaron un total de 384 pacientes de los que 341 presentaban un aclaramiento de creatinina entre 10-50 ml/min. Se revisaron 2.807 medicamentos prescritos, de éstos, 2.052 no requerían ajuste posológico en insuficiencia renal y 508 estaban correctamente ajustados. 247 prescripciones eran susceptibles de un ajuste posológico de las cuales 164 prescripciones era necesario un ajuste posológico concreto. Se realizaron recomendaciones posológicas en 200 ocasiones, y se aceptaron un total de 131. Los fármacos con mayor número de intervenciones fueron la enoxaparina, levofloxacino, amoxicilina-clavulánico y digoxina. Conclusiones: La implantación del programa de atención farmacéutica ha tenido una buena aceptación entre los facultativos prescriptores, siendo los antibióticos el grupo farmacológico más susceptible de realizar un ajuste posológico sobre una mayoría de pacientes con insuficiencia renal moderada (AU)

Background and objective: To asses the outcomes of posological adjust program in renal impairment inpatients, describe the drugs more usually involved, and determine the degree of acceptance of the pharmaceutical intervention made. Material and method: A fifteen months-prospective study, in renal insufficiency inpatients (serum creatinine > 1,4 mg/dL) treated with drugs that needs posological adjustment. The primary outcome was the ratio of adequate dosage of the treatment, according with the glomerular filtration rate. We also evaluated the global acceptation rate, the drugs inadequate prescribed more frequently, phamacokinetic analysis derived from the pharmaceutical intervention and its educative character. Results: 384 patients were identified, and 341 of them presented a glomerular filtration rate between 10-50 ml/min. 2.807 prescribed drugs were reviewed, and 2.052 of them didn’t require posological adjustment in renal insufficiency, 508 prescribed drugs were correctly adjusted. 247 pres - criptions were susceptible of posological adjustment and 164 of them, needed a concrete posological adjustment. We performed 200 posological recommendations, and 131 were accepted. The drugs with a higher number of interventions were enoxaparine, levofloxacin, amoxicillin-clavulanic and digoxin. Conclusions: The implementation of the pharmaceutical care program was accepted between physicians, being antibiotics the group more susceptible of doing a posological adjustment in most patients with renal impairment (AU)

[Pharmaceutical care in renal insufficiency inpatients]. / Atención farmacéutica en pacientes ingresados con insuficiencia renal.

BACKGROUND AND OBJECTIVE: To asses the outcomes of posological adjust program in renal impairment inpatients, describe the drugs more usually involved, and determine the degree of acceptance of the pharmaceutical intervention made. MATERIAL AND METHOD: A fifteen months-prospective study, in renal insufficiency inpatients (serum creatinine > 1,4 mg/dL) treated with drugs that needs posological adjustment. The primary outcome was the ratio of adequate dosage of the treatment, according with the glomerular filtration rate. We also evaluated the global acceptation rate, the drugs inadequate prescribed more frequently, phamacokinetic analysis derived from the pharmaceutical intervention and its educative character. RESULTS: 384 patients were identified, and 341 of them presented a glomerular filtration rate between 10-50 ml/min. 2.807 prescribed drugs were reviewed, and 2.052 of them didn%#39;t require posological adjustment in renal insufficiency, 508 prescribed drugs were correctly adjusted. 247 pres - criptions were susceptible of posological adjustment and 164 of them, needed a concrete posological adjustment. We performed 200 posological recommendations, and 131 were accepted. The drugs with a higher number of interventions were enoxaparine, levofloxacin, amoxicillin-clavulanic and digoxin. CONCLUSIONS: The implementation of the pharmaceutical care program was accepted between physicians, being antibiotics the group more susceptible of doing a posological adjustment in most patients with renal impairment.

Identification of antigenic variants of the porcine rubulavirus in sera of field swine and their seroprevalence.

We sampled sera from 1013 non-vaccinated swine from four states in Mexico, Guanajuato, Jalisco, Michoacán and the Estado de Mexico, to analyse anti-porcine rubulavirus antibody titres against three different porcine rubulavirus isolates (PAC-4/1993, PAC-6/2001, and PAC-9/2003) using a hemagglutination inhibition assay. The results revealed that there were antigenic differences among the isolates assessed. In particular, the estimated correlation between the PAC-4/1993 and PAC-6/2001 (0.50) isolates and between the PAC-4/1993 and PAC-9/2003 isolates (0.56) displayed a moderate positive correlation. In contrast, there was a strong positive correlation between the PAC-6/2001 and PAC-9/2003 isolates (0.73). We also found that in the state of Guanajuato, PAC-4/1993 was the isolate that was most frequently identified; in Jalisco, the isolate was PAC-6/2001; and in Michoacán, the isolate was PAC-9/2003. By contrast, in the Estado de Mexico, all three isolates appeared to circulate with a low seroprevalence. In general, the analysed sera from the four states displayed a porcine rubulavirus serological prevalence ranging from 9% to 23.7%. These data indicate that there is not complete antibody cross-antigenicity among the three isolates, and the antigenic variations in the antibody response found in this study implies that the use of a monovalent vaccine would not generate complete protection against the different antigenic subtypes.

Enterotoxigenic gene profiles of Bacillus cereus and Bacillus megaterium isolates recovered from honey.

One hundred and thirty two Bacillus cereus and 52 Bacillus megaterium isolates from honeys were evaluated for the presence of genes encoding enterotoxin HBL, enterotoxin-T, cytotoxin K and the NHE complex, respectively. The relationship between hemolytic and coagulase activity and its correlation with the presence of the four mentioned enterotoxins was determined by principal component analysis (PCA). PCA in B. cereus revealed a positive correlation among free coagulase, hemolysis and the presence of genes hblA, hblB, hblC, hblD (HBL complex) and bceT (enterotoxin-T), but no correlation with the clumping factor (bound coagulase) and the presence of sequences of the NHE complex. On the other hand, PCA in B. megaterium showed a high positive correlation between coagulase (bound and free) and the haemolytic activity but no correlation in relation to the presence of genes of the HBL complex, cytotoxin K, enterotoxin T and the NHE complex. To our knowledge, this is the first report of the detection of cytotoxin K and of the NHE complex genes in B. megaterium. The relationship between the coagulase activity and the presence of virulence factors has not been described before in the genus Bacillus, being this work the first report of this correlation. Interestingly, the presence of the cytK gene was almost independent of the presence of the rest of virulence factors herein analyzed both in B. cereus and B. megaterium populations. Our results suggest that honey could be a possible vehicle for foodborne illness due to the presence of toxigenic B. cereus and B. megaterium strains containing different virulence factors.

Enterotoxigenic gene profiles of Bacillus cereus and Bacillus megaterium isolates recovered from honey / Búsqueda de factores de virulencia en cepas de Bacillus cereus y de Bacillus megaterium aisladas de miel

One hundred and thirty two Bacillus cereus and 52 Bacillus megaterium isolates from honeys were evaluated for the presence of genes encoding enterotoxin HBL, enterotoxin-T, cytotoxin K and the NHE complex, respectively. The relationship between hemolytic and coagulase activity and its correlation with the presence of the four mentioned enterotoxins was determined by principal component analysis (PCA). PCA in B. cereus revealed a positive correlation among free coagulase, hemolysis and the presence of genes hblA, hblB, hblC, hblD (HBL complex) and bceT (enterotoxin-T), but no correlation with the clumping factor (bound coagulase) and the presence of sequences of the NHE complex. On the other hand, PCA in B. megaterium showed a high positive correlation between coagulase (bound and free) and the haemolytic activity but no correlation in relation to the presence of genes of the HBL complex, cytotoxin K, enterotoxin T and the NHE complex. To our knowledge, this is the first report of the detection of cytotoxin K and of the NHE complex genes in B. megaterium. The relationship between the coagulase activity and the presence of virulence factors has not been described before in the genus Bacillus, being this work the first report of this correlation. Interestingly, the presence of the cytK gene was almost independent of the presence of the rest of virulence factors herein analyzed both in B. cereus and B. megaterium populations. Our results suggest that honey could be a possible vehicle for foodborne illness due to the presence of toxigenic B. cereus and B. megaterium strains containing different virulence factors.

Se evaluaron 132 aislamientos de Bacillus cereus y 52 de Bacillus megaterium provenientes de mieles de distintos orígenes geográficos para investigar la presencia de secuencias de ADN relacionadas con genes de virulencia y su posible correlación con la actividad hemolítica y coagulasa. Con respecto a los genes de virulencia, se analizaron por PCR secuencias de ADN de los genes nhe (A, B y C), HBL (A, B, C, D), cytK y bceT. La relación entre las variables fue evaluada mediante un análisis de componentes principales, donde se encontró que los aislamientos de B. cereus mostraron una correlación positiva entre actividad de coagulasa (coagulasa libre) y presencia de los genes del complejo HBL y bceT, mientras que en B. megaterium se halló una alta correlación positiva entre actividad de coagulasa (libre y fija) y actividad hemolítica, pero no se observó correlación significativa entre la presencia de genes de virulencia y dichas actividades. Este estudio constituye el primer registro de la presencia de los genes cyt K y NHE en cepas de B. megaterium y el primer trabajo que analiza la relación entre la actividad de coagulasa y la presencia de genes de virulencia en B. cereus y B. megaterium. La presencia del gen cytK en ambas especies resultó totalmente independiente del resto de los factores de virulencia analizados. Nuestros hallazgos sugieren que la miel podría vehiculizar enfermedades transmisibles por alimentos debido a la presencia de cepas de B. cereus y B. megaterium potencialmente tóxicas.

First Report of Agrobacterium rubi and A. rhizogenes Causing Crown and Root Gall and Hairy Root on Blueberry in Argentina.

From 2006 to 2009, crown gall and hairy root symptoms were observed on blueberry (Vaccinium corymbosum cvs. O'Neil, Millennia, and Misty) plants from six nurseries in Tucumán, Concordia, Pilar, Morón, and Baradero, Argentina. Bacteria were isolated from galls of all three cultivars and from hairy roots of Millenia and O'Neil onto D1 and D1M agar media at 27°C. Typical Agrobacterium colonies developed in 5 days (2). Seven bacterial strains (five from galls and two from hairy roots) were studied further. All were gram negative, aerobic, and catalase positive with rod-shaped cells that synthesized ß-galactosidase and metabolized D-glucose, D-arabinose, n-acetyl-glucosamine, maltose, mannitol, and malonate. Strains were negative for lysine decarboxylase, H2S production, indole, and 3-ketolactose production. While gall strains were urease positive and citrate variable (mostly positive), hairy root strains were urease negative, citrate positive, had poly-ß-hydroxybutyrate inclusion granules, and clarified acid on potato dextrose agar containing 0.5% CaCO3 (2). Agrobacterium tumefaciens ATCC 15955 and LBA 958 were included as controls. PCR with virA/C primers amplified a 338-bp product corresponding to the virD2 operon and confirmed that the strains harbored a pathogenic plasmid (1). Bacterial strains were assigned to biovars with a multiplex PCR assay targeting 23S rRNA sequences (3). Two strains produced PCR amplicons typical of A. rhizogenes bv. 2. The other five strains produced PCR amplicons typical of A. rubi, which were insensitive to agrocin in a bioassay with A. radiobacter strain K1026. Identity was confirmed by sequencing the 16S rDNA of strains F 266 (GenBank No. GU580894) and F 289 (No. GU580895), which had 99% homology to 16sRNA sequences of A. rubi ICMP 11833 (AY626395.1) and A. rhizogenes ATCC 11325 (AY945955.1), respectively. Pathogenicity of all seven strains was tested on V. corymbosum cv. Misty, Bryophyllum daigremontiana, tobacco cv. Xanthi, tomato cv. Presto, and pepper cv. California Wonder. Plants were inoculated by a needle stabbed into the stems with the appropriate cell suspension (108 CFU/ml) of each strain or with sterile distilled water (control treatment). Two plants of each species were tested per strain. Plants were grown for at least 45 days at 23 ± 3°C and symptoms were recorded. Inoculations with the five strains isolated from galls caused development of spherical, white to flesh-colored, rough, spongy wart-like galls at the inoculation sites. Root strains induced root proliferation on all inoculated plants as well as in a carrot disk bioassay (4). On blueberry plants, galls were dark brown to black, rough, and woody 6 months after inoculation. No lesions were observed on control plants. Bacteria were reisolated from symptomatic tissues of inoculated plants. Enterobacterial repetitive intergeneric consensus-PCR confirmed that the DNA fingerprints of the reisolated strains were identical to those of the original strains. To our knowledge, this is the first report of A. rubi and A. rhizogenes causing hairy root and crown gall on blueberry in Argentina. References: (1) J. H. Haas et. al. Appl. Environ. Microbiol. 61:2879,1995. (2) L. W. Moore et al. Page 17 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. N. W. Schaad et al., eds. The American Phytopathological Society, St. Paul, MN, 2001. (3) J. Pulawska et al. Syst. Appl. Microbiol. 29:470, 2006. (4) M. H. Ryder et al. Plant Physiol. 77:215, 1985.

First Report of Leaf Spot Disease of Maize Caused by Pantoea ananatis in Argentina.

From 2007 to 2008, an uncharacterized disease of maize (Zea mays L.) was observed in commercial fields of Laguna Blanca, Formosa, Argentina and from different fields of Santa Fe and Catamarca provinces of Argentina. Symptoms included light-colored necrotic streaks on leaves and tan or white irregular blotches that sometimes were surrounded by reddish purple-to-dark brown margins. Severity of symptoms varied greatly from one field to another. Abundant bacterial streaming was observed from lesions when examined at ×150. Gram-negative, facultatively anaerobic bacteria were consistently isolated from lesions. These formed light yellow-to-orange, glistening, convex colonies on yeast dextrose calcium carbonate agar incubated at 30°C. Ten isolates from ten different symptomatic plants were selected for further study. All isolates were motile, induced a hypersensitive response in tobacco plants, and were oxidase negative. Colonies developed at 37°C. Physiological and biochemical characterization with the API 20E test strips and database (bioMerieux, Buenos Aires, Argentina) showed that the strains belonged to the genus Pantoea. All strains were positive for ß-galactosidase, utilized citrate and tartrate, and produced acid from d-glucose, d-mannitol, d-melibiose, l-arabinose, sucrose, meso-inositol, glycerol, d-sorbitol, and amygdalin. All were negative for arginine dihydrolase, lysine decarboxylase, ornithine decarboxylase, tryptophane deaminase, H2S production, urease, and reduction of nitrate to nitrite. Variable results were obtained for indole, gelatinase, and l-rhamnose. Their identity was confirmed by sequencing the 16S rRNA gene strain F327 (GenBank Accession No. GU068363). A BlastN search of GenBank revealed 99% nt identity with strains LMG 20103 (AF364847.1), LMG 20105 (AF364845.1), and LMG 2665 (FJ611815.1) of Pantoea ananatis. Pathogenicity was verified on Z. mays (EM 6079 HX, Dow Morgan) by injection-infiltration of bacterial suspensions at 105 CFU/ml. Controls were infiltrated with sterile distilled water. Plants were kept at 26 ± 3°C in a greenhouse. Symptoms were first detected 15 to 17 days after inoculation and then lesions expanded to resemble natural infections within 30 days. Bacteria were reisolated and the original and reisolated strains were compared by using repetitive sequence-based (rep)-PCR with ERIC primers (1) and fingerprints of the reisolated strains were identical to those of the original strains, thereby fulfilling Koch's postulates. No lesions were observed on controls. Known strains of P. stewartii from the United States (SW2, DC400, DC441, and DC283) were also tested for comparison. On the basis of sequencing data, pathogenicity, and physiological tests, the pathogen was identified as P. ananatis (4). To our knowledge, this is the first report of P. ananatis causing a disease of maize in Argentina, although a similar disease has been reported in Brazil (2) and Mexico (3). References: (1) F. J. Louws et al. Appl. Environ. Microbiol. 60:2286, 1994. (2) L. D. Paccola-Meirelles et al. J. Phytopathol. 149:275, 2001. (3) R. Pérez-y-Terrón et al. Australas. Plant Dis. Notes 4:96, 2009. (4) N. W. Schaad et al., eds. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001.

First Report of Leaf Spot of Dieffenbachia picta and Aglaonema commutatum Caused by Burkholderia gladioli in Argentina.

During May of 2008 (austral autumn), an uncharacterized disease was observed on Dieffenbachia picta (Lodd.) Schott and Aglaonema commutatum Schott in commercial greenhouses in Pontevedra (34°45'6″S, 58°42'42″W), Argentina. Affected plants showed irregular, brown lesions on leaves, approximately 15 to 20 mm in diameter, surrounded by water-soaked haloes that progressed inward from the margins. Water-soaked rotting symptoms were also observed in petioles. Disease incidence approached 80%. Abundant bacterial streaming was observed from lesions when examined at ×100. Bacteria consistently isolated from lesions formed cream-colored, glistening, convex colonies on sucrose peptone agar and produced a yellowish green, diffusible, nonfluorescent pigment on King's medium B. Four isolates from different symptomatic plants were selected for further study. All were aerobic, gram-negative rods that accumulated poly-ß-hydroxybutyrate inclusions. In LOPAT tests, all induced a hypersensitive response in tobacco plants, caused soft rot of potato tubers, and were positive for levan, negative for arginine dihydrolase, and variable for oxidase. All isolates oxidized glucose, did not hydrolyze starch and were able to rot onion slices. Colonies developed at 41°C but not at 4°C. With the API 20NE test strips and database (bioMerieux, Buenos Aires, Argentina), all isolates matched (99% identity) Burkholderia cepacia, but their inability to metabolize cellobiose and sucrose further identified them as B. gladioli. For molecular identification, 23S rDNA was amplified by PCR using B. gladioli-specific primers LP1 and LP4, which yielded a 700-bp product (3), and PCR-restriction fragment length polymorphism of 16S rDNA using AluI (2). PCR products were identical to those from the type strain for B. gladioli, ICMP 3950, isolated from Gladiolus spp. that had been included in all tests for comparison. Pathogenicity was verified on D. picta and A. commutatum by spraying the plants with bacterial suspensions in sterile distilled water at 108 CFU/ml with and without wounding the leaves with a sterile needle and also by injection-infiltration of bacterial suspensions at 105 CFU/ml. In addition, another host plant, Gladiolus communis L., was inoculated in the same manner. Controls were sprayed or infiltrated with sterile distilled water. After 48 h in a humidity chamber, plants were kept at 25 ± 3°C in a greenhouse. In all hosts, symptoms were first detected 3 days after inoculation and lesions expanded to resemble natural infections within 4 to 7 days. All strains caused necrosis around the inoculation sites and lesions were identical to those induced by the ICMP reference strain. Bacteria were reisolated from each host tested and then the original and reisolated strains were compared by enterobacterial repetitive intergeneric consensus-PCR (1); DNA fingerprints of the reisolated strains were identical to those of the original strains, thereby fulfilling Koch's postulates. No lesions were observed on controls or on plants inoculated by spraying without wounding, suggesting that bacteria gain entry through wounds. On the basis of PCR and physiological tests the pathogen was identified as B. gladioli (2-4). To our knowledge, this is the first report of B. gladioli on Dieffenbachia and Aglaonema spp. References: (1) F. J. Louws et al. Appl. Environ. Microbiol. 60:2286, 1994. (2) C. Van Pelt et al. J. Clin. Microbiol. 37:2158, 1999. (3) P. W. Whitby et al. J. Clin. Microbiol. 38:282, 2000. (4) E. Yabuuchi et al. Microbiol. Immunol. 36:1251, 1992.

First Report of Pectobacterium carotovorum subsp. carotovorum on Spathiphyllum wallisii in Argentina.

Peace lily (Spathiphyllum wallisii Regel) is a popular ornamental potted plant in Argentina. During May of 2008 (austral autumn), necrotic lesions of unknown etiology were observed on S. wallisii in a nursery in Pontevedra (34°45'6″S, 58°42'42″W). Plants first showed water-soaked areas starting from the leaf tips. Infected tissue became irregular, brown, dark-to-black lesions on leaves ~12 to 14 mm in diameter surrounded by yellowish haloes. Disease incidence approached 30%. Abundant bacterial streaming was observed from lesions when examined at ×100. Bacteria isolated from lesions formed white-to-cream, glistening, convex colonies on yeast dextrose calcium carbonate agar. Three bacterial strains isolated from different symptomatic plants were selected for comparative analysis with Pectobacterium carotovorum subsp. carotovorum type strain ATCC 15713. All were facultatively, anaerobic, gram-negative rods, pectolytic on crystal violet pectate agar, nonfluorescent on King's medium B, and elicited a hypersensitive response in tobacco plants. All strains were oxidase and arginine dihydrolase negative, fermented glucose, did not hydrolyze starch, did not produce lecithinase, indole or the blue pigment indigoidine, reduced nitrates, hydrolyzed gelatin and esculin, able to rot onion slices, caused soft rot of potato tubers, resistant to erythromycin, and grew at 37°C. Acid was produced from cellobiose, d-glucose, d-melibiose, d-mannitol, d-mannose, l-rhamnose, d-sucrose, and l-arabinose but not from inositol and d-sorbitol. Bacteria utilized N-acetyl-glucosamine and citrate but not tartrate, benzoate, or propionate. Their identity was confirmed by 16S rRNA gene sequencing of strain F402Pcc (GenBank Accession No. FJ717337) showing a 99% homology with that of strain ATCC 3326 (FJ 5958691). Pathogenicity was verified on S. wallisii, Dieffenbachia picta, Aglaonema commutatum, and Anthurium andraeanum within the Araceae family by spraying two plants per strain tested with bacterial suspensions (108 CFU/ml) in sterile distilled water with and without wounding the leaves with sterile needles. Controls were sprayed with sterile distilled water. After 48 h in a humidity chamber, inoculated plants and controls were maintained at 25 ± 3°C in a greenhouse. Water-soaked areas developed from 24 to 48 h after inoculation and became necrotic within 4 to 5 days. Lesions expanded to resemble natural infection in S. wallisii within 20 days, while in the rest of the hosts tested, lesions were smaller and remained brown surrounded by yellowish haloes. All strains were reisolated from each host tested. The original and all reisolated strains were compared by enterobacterial repetitive intergeneric consensus-PCR (4) confirming that DNA fingerprints of the reisolated strains were identical to those of the original strains. No lesions were observed on controls. The pathogen was identified as P. carotovorum subsp. carotovorum based on biochemical, physiological, pathogenicity tests, and 16S rRNA sequencing (1-3).To our knowledge, this is the first report of this pathogen on S. wallisii in Argentina although it has been reported as causing tomato pith necrosis (1) and soft rot of vegetables after harvest (3). References: (1) A. M. Alippi et al. Plant Dis. 81:230, 1997. (2) L. Gardan et al. Int. J. Syst. Evol. Microbiol. 53:381, 2003. (3) L. Halperin and L. S. Spaini. Rev. Argent. Agron. 6:261, 1939. (4) F. J. Louws et al. Appl. Environ. Microbiol. 60:2286, 1994.

Tetracycline and oxytetracycline resistance determinants detected in Bacillus cereus strains isolated from honey samples / Detección de determinantes de resistencia a tetraciclina y oxitetraciclina en cepas de Bacillus cereus aisladas de muestras de miel

The aim of this study was to investigate the presence of tetracycline and oxytetracycline resistance determinants in Bacillus cereus strains isolated from honey samples. Of a total of 77 isolates analyzed, 30 (39%) exhibited resistance to tetracyclines according to the results of a disk diffusion method. Resistant strains (n=30) were screened by PCR for the presence of the resistant determinants tetK, tetL, tetM, tetO, tetW, otrA and otrB and their MIC values for tetracycline, oxytetracycline and minocycline were assessed. According to the PCR results, 23 isolates (77%) presented at least one tetracycline or oxytetracycline resistance determinant. The tetK genotype was present in 10 isolates while the tetL, tetM, and otrA genotypes were present in 3, 2, and 5 isolates, respectively. In addition, 2 isolates of the tetK plus tetM genotype, 1 of the tetK plus tetL genotype, and 1 of the tetK plus otrA genotype were found. All isolates were tetW, tetO and otrB negatives. On the other hand, 7 isolates (23%) showed a tetracycline-resistant and/or minocyclineresistant phenotype (MIC) but did not carry any of the tet or otr determinants investigated in this study. This research has shown that B. cereus isolates from honey samples contain a variety of tetracycline and oxytetracycline resistance genes, including the tetK and tetL determinants which encode for efflux proteins, and tetM and otrA, which encode for ribosomal protection proteins. These findings indicate that strains isolated from honeys could represent a reservoir for tetracycline resistance genes. To our knowledge, this is the first report of tetracycline-resistant and oxytetracyclineresistant B. cereus strains carrying the tetK determinant, and also the first report of oxytetracycline-resistant and tetracycline- resistant Bacillus species carrying the otrA determinant.

El objetivo del presente estudio ha sido investigar la presencia de diversos determinantes de resistencia a tetraciclina y oxitetraciclina en las poblaciones de Bacillus cereus presentes en la miel. De un total de 77 aislamientos evaluados, 30 (39%) resultaron resistentes a tetraciclina y/o minociclina de acuerdo con los resultados de las pruebas de difusión en disco. Dentro del grupo que presentó un fenotipo resistente, se investigó la presencia de los determinantes tetK, tetL, tetM, tetO, tetW, otrA y otrB por PCR y se determinaron los valores de CIM para tetraciclina, oxitetraciclina y minociclina. De acuerdo con los resultados obtenidos por PCR, 23 aislamientos (77%) presentaron al menos un determinante de resistencia a tetraciclina o a oxitetraciclina; el genotipo tetK se encontró en 10 de esos aislamientos, mientras que los genotipos tetL, tetM y otrA se hallaron en 3, 2 y 5 aislamientos, respectivamente. Ningún aislamiento presentó los genotipos tetW, tetO ni otrB. Adicionalmente, se encontraron los genotipos tetK plus tetM (2 aislamientos); tetK plus tetL (1 aislamiento) y tetK plus otrA (1 aislamiento). Por otra parte, 7 cepas (23%) resultaron resistentes a tetraciclina, oxitetraciclina y/o minociclina por CIM, pero no presentaban ninguno de los determinantes tet u otr estudiados. Estos resultados indican la existencia de un alto porcentaje de cepas de B. cereus aisladas de miel con genes de resistencia a tetraciclina y oxitetraciclina, incluyendo los determinantes tetK, tetL, tetM y otrA. Este estudio constituye el primer registro de la presencia del determinante tetK de resistencia a tetraciclina en B. cereus, como así también la presencia del determinante otrA dentro del género Bacillus.

Tetracycline and oxytetracycline resistance determinants detected in Bacillus cereus strains isolated from honey samples.

The aim of this study was to investigate the presence of tetracycline and oxytetracycline resistance determinants in Bacillus cereus strains isolated from honey samples. Of a total of 77 isolates analyzed, 30 (39%) exhibited resistance to tetracyclines according to the results of a disk diffusion method. Resistant strains (n=30) were screened by PCR for the presence of the resistant determinants tetK, tetL, tetM, tetO, tetW, otrA and otrB and their MIC values for tetracycline, oxytetracycline and minocycline were assessed. According to the PCR results, 23 isolates (77%) presented at least one tetracycline or oxytetracycline resistance determinant. The tetK genotype was present in 10 isolates while the tetL, tetM, and otrA genotypes were present in 3, 2, and 5 isolates, respectively. In addition, 2 isolates of the tetK plus tetM genotype, 1 of the tetK plus tetL genotype, and 1 of the tetK plus otrA genotype were found. All isolates were tetW, tetO and otrB negatives. On the other hand, 7 isolates (23%) showed a tetracycline-resistant and/or minocycline-resistant phenotype (MIC) but did not carry any of the tet or otr determinants investigated in this study. This research has shown that B. cereus isolates from honey samples contain a variety of tetracycline and oxytetracycline resistance genes, including the tetK and tetL determinants which encode for efflux proteins, and tetM and otrA, which encode for ribosomal protection proteins. These findings indicate that strains isolated from honeys could represent a reservoir for tetracycline resistance genes. To our knowledge, this is the first report of tetracycline-resistant and oxytetracycline-resistant B. cereus strains carrying the tetK determinant, and also the first report of oxytetracycline-resistant and tetracycline-resistant Bacillus species carrying the otrA determinant.

Geraldton Wax as a New Host for Agrobacterium tumefaciens in Argentina.

During 2004, Geraldton wax plants (Chamaelaucium uncinatum cv. BM Violet) from commercial greenhouses in La Plata, Argentina showed gall-like structures on collars and roots similar to those reported by Carsten et al. (2). No pathogenic fungi were associated with lesions. Bacteria isolated from galls grown on medium 1A and D1M agar yielded colonies typical of Agrobacterium sp. (4). Four isolates were selected for further study. All isolates were aerobic, gram-negative rods and produced 3-ketolactose, but did not produce alkali from l-tartrate or accumulate poly-ß-hydroxybutyrate inclusions. The isolates were able to grow with 2% NaCl, at 35°C, and also on potato dextrose agar medium containing CaCO3 (4) but without acid clarification. Polymerase chain reaction analysis with primers VCF/VCR that amplified the expected 730-bp product of virC operon confirmed that all strains harboured a Ti plasmid (3,4). In addition, strains were screened for extrachromosomal DNA by the in-well lysis and electrophoresis procedure of Eckhardt with minor modifications as reported by Albiach and Lopez (1) and compared with strain Agrobacterium tumefaciens LBA 958, all Argentinean strains tested harbored a plasmid similar in size. Pathogenicity was verified on Geraldton wax and tobacco plants. Bacterial suspensions of each isolate (108 CFU/ml) were pricked into the stems. Control plants were pricked with sterile distilled water. Plants were maintained at 23 ± 3°C and symptoms were recorded after 45 days. Development of almost spherical, white-to-flesh-colored, rough, spongy and wart-like galls at the inoculation sites of the inoculated collars, stems, and trunks were registered. In Geraldton wax, as galls aged, they become dark brown to black, rough, and woody. Bacteria were reisolated from these galls fulfilling Koch's postulates. No lesions were observed on the controls. To our knowledge, this is the first report of A. tumefaciens on C. uncinatum in Argentina. References: (1) M. R. Albiach and M. M. López. Appl. Environ. Microbiol. 58:2683, 1992. (2) E. Carstens et al. Plant Dis. 83:783, 1999. (3) H. Sawada et al. Appl. Environ. Microbiol. 61:828, 1995. (4) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society. St. Paul, MN, 2001.

Pegylated interferon-alpha 2b-ribavirin combination in Egyptian patients with genotype 4 chronic hepatitis.

Egypt has a high prevalence rate of hepatitis C (HCV) infection and as much as 90% is genotype 4. Response to interferon (IFN) varies with viral genotype and degree of fibrosis. Genotype 4 is poorly sensitive to standard IFN and IFN-ribavirin combination. We evaluated pegylated interferon (PEG-IFN)-alpha2b in our patients. Sixty-one patients with compensated chronic HCV genotype 4 were enrolled in two groups: group A (31 patients) received IFN-alpha2b 3 MU three times per week and group B (30 patients) received 1.5 mug/kg PEG-IFN-alpha2b once weekly. Ribavirin was added to each regimen in a dose of 800-1200 mg based on body weight. Patients were followed up for 24 weeks to assess the sustained response (SR). End-of-treatment response (ETR) was achieved in 11 of 31 patients (35.48%) in group A, and 13 of 30 patients (43.33%) in group B (P < 0.05). Only eight patients in group A and 10 in group (B) achieved a sustained virological response (25.8 and 33.3%, respectively) (P < 0.05). By computing ETR, SR or relapse and pretreatment baseline data (pretreatment, viral load, alanine transaminases, necroinflammatory and hepatic fibrosis), both inter- and intragroup, no significant correlations could be detected. In terms of safety and tolerability, PEG-IFN-alpha2b and IFN-alpha2b were comparable. In spite of mild insignificant increase in ETR and SR with the pegylated form, the poor response of genotype 4 in Egypt (genotype 4a) to different forms of IFNs may be related to an intrinsic resistance to the direct antiviral effect of IFN.

Developmental differences between cation-independent and cation-dependent mannose-6-phosphate receptors in rat brain at perinatal stages.

Mannose-6-phosphate receptors (MPRs) play a role in the selective transport of macromolecules bearing mannose-6-phosphate residue to lysosomes. To date, two types of MPRs have been described in most of cells and tissues: the cation-dependent (CD-MPR) and cation-independent mannose-6-phosphate receptor (CI-MPR). In order to elucidate their possible role in the central nervous system, the expression and binding properties of both MPRs were studied in rat brain along perinatal development. It was observed that the expression of CI-MPR decreases progressively from fetuses to adults, while the CD-MPR increases around the 10th day of birth, and maintains these values up to adulthood. Binding assays showed differences in the Bmax and KD values between the ages studied, and they did not correlate with the expression levels of both MPRs. Variations in lysosomal enzyme activities and expression of phosphomannosylated ligands during development correlated more with CD-MPR than with CI-MPR expression. These results suggest that both receptors play a different role in rat brain during perinatal development, being CD-MPR mostly involved in lysosome maturation.

Fetal lung volumetry using two- and three-dimensional ultrasound.

OBJECTIVES: To compare methods of measuring fetal pulmonary volume and to establish nomograms of fetal pulmonary volume according to gestational age for the accurate diagnosis of pulmonary hypoplasia. METHODS: Three methods of measuring fetal pulmonary volume in 39 normal fetuses were compared: two-dimensional (2D) ultrasound measurement assuming that the lung is a geometrical pyramid, three-dimensional (3D) ultrasound using the VOCAL rotational method, and the conventional multiplanar 3D mode. Linear regression was used to construct an equation for 3D volume calculation from 2D measurements (the re-evaluated pulmonary volume equation (RPVE)). Lung volume measurements were recorded from 622 singleton fetuses in order to construct nomograms. RESULTS: There was no statistically significant difference between the lung volume values obtained using the two 3D modes. However, in comparison with the 2D measurements the volumes obtained were larger (mean difference = 11.99, P < 0.1 x 10(-6)). The relationship between the 2D and 3D volumes was determined using a statistical linear regression method: RPVE (mL) = 4.24 + (1.53 x 2DGPV), where 2DGPV (2D geometric pulmonary volume) = (surface area right lung base (cm2) + surface area left lung base (cm2)) x 1/3 height right lung (cm). Two nomograms were constructed, one for use with 2D and one for 3D technology. CONCLUSION: 2D pulmonary volume assessment can be used in clinical situations where fetal prognosis depends on lung volume and its growth potential. It is routinely available and easy to perform particularly when repeat measurements are required in evaluation of lung growth. We therefore propose this method as an alternative to magnetic resonance imaging or 3D ultrasound.