Discrepancies on quality perceived by the patients versus professionals on the quality of a nuclear medicine department.

PURPOSE: To evaluate the discrepancies between the professionals and outpatients on quality perceived of a Nuclear Medicine Department (NMD). MATERIAL AND METHODS: This cross-sectional study has been carried out using two questionnaires: a validated patient experience questionnaire and a quality perception questionnaire for professionals. Both questionnaires use the same 25 categorical items to measure service quality, 2 Likert scale items to measure satisfaction and willingness to recommend the NMD and 1 open-ended question. The patient questionnaire included 6 socio-demographic items and one job-related question (professionals). The categorical items were classified as "conformity" or "non-conformity." RESULTS: The response rate was 36.7% for outpatients and 100% for professionals. Mean value for satisfaction with the NMD was 9 points for patients and 6.9 points for professionals. Mean number of non-conformity items per person was 2.8 for the patient group and 8.7 for the professional group. Cohen's Kappa value was 0.112, indicating poor agreement in the classification of items as strong points and areas for improvement. Of the 25 items, the professionals and patients coincided on 12 (48%). CONCLUSION: Agreement was low between the quality perception of patients and professionals. The patients scored quality of service higher than the NMD professionals did. These instruments are useful aid to help health organizations detect areas for improvement, and to improve the quality of the service provided to patients.

[Personalizing the reference level: gold standard to evaluate the quality of service perceived]. / Personalización del nivel de referencia: patrón oro para evaluar la calidad de servicio percibida.

OBJECTIVE: To know the cutoff point at which in-house Nuclear Medicine Department (MND) customers consider that the quality of service is good (personalized cutoff). MATERIAL AND METHOD: We conducted a survey of the professionals who had requested at least 5 tests to the Nuclear Medicine Department. A total of 71 doctors responded (response rate: 30%). A question was added to the questionnaire for the user to establish a cutoff point for which they would consider the quality of service as good. The quality non-conformities, areas of improvement and strong points of the six questions measuring the quality of service (Likert scale 0 to 10) were compared with two different thresholds: personalized cutoff and one proposed by the service itself a priori. Test statistics: binomial and Student's t-test for paired data. RESULTS: A cutoff value of 7 was proposed by the service as a reference while 68.1% of respondents suggested a cutoff above 7 points (mean 7.9 points). The 6 elements of perceived quality were considered strong points with the cutoff proposed by the MND, while there were 3 detected with the personalized threshold. Thirteen percent of the answers were nonconformities with the service cutoff versus 19.2% with the personalized one, the differences being statistically significant (difference 95% CI 6.44%:0,83-12.06). CONCLUSIONS: The final image of the perceived quality of an in-house customer is different when using the cutoff established by the Department versus the personalized cutoff given by the respondent.

[Evaluation of the efficacy of sentinel node detection in breast cancer: chronological course and influence of the incorporation of an intra-operative portable gamma camera]. / Evaluación de la eficacia en la detección del ganglio centinela en el cáncer de mama: evolución cronológica e influencia de la incorporación de una gammacámara portátil intraoperatoria.

AIM: To define the sentinel node identification rate in breast cancer, the chronological evolution of this parameter and the influence of the introduction of a portable gamma camera. MATERIAL AND METHODS: A retrospective study was conducted using a prospective database of 754 patients who had undergone a sentinel lymph node biopsy between January 2003 and December 2011. The technique was mixed in the starting period and subsequently was performed with radiotracer intra-peritumorally administered the day before of the surgery. Until October 2009, excision of the sentinel node was guided by a probe. After that date, a portable gamma camera was introduced for intrasurgical detection. RESULTS: The SN was biopsied in 725 out of the 754 patients studied. The resulting technique global effectiveness was 96.2%. In accordance with the year of the surgical intervention, the identification percentage was 93.5% in 2003, 88.7% in 2004, 94.3% in 2005, 95.7% in 2006, 93.3% in 2007, 98.8% in 2008, 97.1% in 2009 and 99.1% in 2010 and 2011. There was a significant difference in the proportion of identification before and after the incorporation of the portable gamma camera of 4.6% (95% CI of the difference 2-7.2%, P = 0.0037). CONCLUSIONS: The percentage of global identification exceeds the recommended level following the current guidelines. Chronologically, the improvement for this parameter during the study period has been observed. These data suggest that the incorporation of a portable gamma camera had an important role.

[Assessment of the patient satisfaction with a Nuclear Medicine Service]. / Evaluación del grado de satisfacción de los pacientes atendidos en un servicio de Medicina Nuclear.

GOAL: To know the perceived quality and the levels of patient satisfaction with the Nuclear Medicine Service (MN). METHODS: A cross-sectional descriptive study was performed. The authors designed a self-applied questionnaire based on a questionnaire from a survey created by the National Health Service of the UK. The answers of 32 items were analyzed, including 4 social-demographic questions and one open question. The authors recoded the variables related to service quality and recorded them as "in accordance" and "not in accordance." The validity of the questionnaire was measured using Cronbach's alpha and determination (R(2)) indexes. The authors used the χ(2), Student's T, ANOVA and linear regression analysis statistical tests. RESULTS: A total of 179 questionnaires were analyzed (response rate: 36.6%, sampling error: 5.8%). Evaluation of general satisfaction and the recommendation of the NM Service obtained a mean score of 8.96 and 9.20 (1-10 scale) points, respectively. The most influential variable regarding general satisfaction was the general impression of the organization of the service. The strong points of the service were courtesy, general organizational image and cleanliness. The main areas for improvement were appointment change process and waiting list. There were no significant differences regarding satisfaction due to the social-demographic variables except for age. CONCLUSION: This satisfaction survey has shown that patients are satisfied with the Nuclear Medicine Service and that it is a useful tool to detect the strong points and areas for improvement of the Service from the user's perspective.

Expression and purification of Z protein from Junín virus.

Arenaviridae comprises 23 recognized virus species with a bipartite ssRNA genome and an ambisense coding strategy. The virions are enveloped and include nonequimolar amounts of each genomic RNA species, designated L and S, coding for four ORFs (N, GPC, L, and Z). The arenavirus Junín (JUNV) is the etiological agent of Argentine Hemorrhagic Fever, an acute disease with high mortality rate. It has been proposed that Z is the functional counterpart of the matrix proteins found in other negative-stranded enveloped RNA viruses. Here we report the optimized expression of a synthetic gene of Z protein, using three expression systems (two bacterial and a baculoviral one). One of these recombinant proteins was used to generate antibodies. A bioinformatic analysis was made where Z was subdivided into three domains. The data presented contributes methodologies for Z recombinant production and provides the basis for the development of new experiments to test its function.

[Sentinel ganglion in breast cancer: selective biopsy compared with axillary lymphadectomy]. / Ganglio centinela en cáncer de mama: biopsia selectiva comparada con linfadenectomía axilar. Seguimiento a largo plazo.

BACKGROUND: Selective biopsy of the sentinel ganglion (SBSG) has replaced axillary lymphadectomy (AL) as the procedure of choice in staging breast cancer in its initial stages and in clinically negative axilla. The aim of this study is to compare global event-free survival of those patients subjected to SBSG followed by AL, during the period of validation of the technique, with respect to those subjected to SBSG and AL if the sentinel ganglion (SG) showed metastasis. METHODS: One hundred and forty-eight patients were included, 81 belonging to the period of validation and 67 to the clinical application group. Radiocoloid was administered intraperitumorally, obtaining images up until the visualisation of the SG; its identification and extirpation were carried out subsequently in the surgical intervention. RESULTS: The efficacy of the technique in the validation group was 92.5%, sensitivity was 95.6% and the rate of false negatives was 4%. Of the 81 patients, 75 are free of disease (92.6%). Of the 67 patients belonging to the clinical application group, 63 (94%) are free of disease. No patient has presented axillary ganglion recurrence. CONCLUSION: In the validation of the technique we obtained values that fall within the demands of generally accepted quality. With an average follow up of 6 years we did not observe axillary ganglion recurrence in any of the two groups. There is no statistically significant difference in global and event free survival between the two groups.

[Purification and breaking techniques for cysts of Giardia spp]. / Técnicas de purificación y ruptura de quistes de Giardia spp.

The purpose of this study was to optimize and evaluate the purification techniques, isolation and breaking of cysts of Giardia spp from fecal samples to isolate DNA. Filtrated fecal samples were tested in 3 purification techniques: Telleman solution, sucrose and Telleman plus sucrose. The sucrose solution let us to isolate the cysts with less detritus. The cleaned cysts were splited in 3 techniques to test the breaking: osmotic shock and heat, chemistry degradation and thermic shock, enzymatic action and mechanic effect. Only the last method was successful and showed bands in agarose gel. The result of this study shows a routine and common method which could be used in the previous steps to the PCR technique for the genotypification of these parasites.

Técnicas de purificación y ruptura de quistes de Giardia spp / Purification and breaking techniques of cysts of Giardia spp

El objetivo de este trabajo fue optimizar y evaluar las técnicas de purificación, aislamiento y ruptura de quistes de Giardia spp a partir de heces formoladas para la obtención de ADN. La materia fecal filtrada fue sometida a 3 técnicas de purificación, utilizando soluciones de formol-éter, sacarosa y formol-éter más sacarosa. La solución de sacarosa permitió aislar los quistes con menos detritos. Los quistes purificados fueron tratados con 3 técnicas para la ruptura de los mismos: shock osmótico y calor, degradación química y shock térmico, acción enzimática y efecto mecánico. Solamente con la técnica de shock térmico, acción enzimática y efecto mecánico se observaron bandas fluorescentes en geles de agarosa. Los resultados de este trabajo permiten contar con una metodología de rutina, simple, que podría ser usada en los pasos previos a la técnica de PCR para la genotipificación de este parásito.

The purpose of this study was to optimize and evaluate the purification techniques, isolation and breaking of cysts of Giardia spp from fecal samples to isolate DNA. Filtrated fecal samples were tested in 3 purification techniques: Telleman solution, sucrose and Telleman plus sucrose. The sucrose solution let us to isolate the cysts with less detritus. The cleaned cysts were splited in 3 techniques to test the breaking: osmotic shock and heat, chemistry degradation and thermic shock, enzymatic action and mechanic effect. Only the last method was successful and showed bands in agarose gel. The result of this study shows a routine and common method which could be used in the previous steps to the PCR technique for the genotypification of these parasites.

[Bacteremia and pseudobacteremia caused by coagulase-negative Staphylococcus in children]. / Bacteriemia y pseudobacteriemia causada por Staphylococcus coagulasa negativa en niños.

OBJECTIVE: To compare patients with Coagulase Negative Staphylococcus bacteremia (CoNS-B) and pseudobacteremia (CoNS-PB) in a pediatric hospital. METHODS: Descriptive and comparative study between children diagnosed with CoNS-B and CoNS-PB. RESULTS: A total of 159 children with CoNS positive blood cultures were evaluated. 66 children were classified as CoNS-B (41.5%) and 93 as CoNS-PE3 (58.5%). On the average CoNS was isolated on day 21 among children with bacteremia (B) and on day 2 in those with PB (p < 0.01). Excluding newborns, patients with B and PB had on average 2.6 and 1.1 positive cultures respectively. Most children with bacteremia were at the intensive care unit (67.2%), while patients with PB were mostly detected at the emergency room. Using logistic regression analysis, we found four factors independently associated with CoNS bacteremia: total parenteral nutrition (OR 5.4; 95% CI 2.2-12.9), low birth weight (OR 2.6; 95% CI 1.1-5.9), catheters placed by cut-down technique (OR 1.9; 95% CI 1.1-3.8), and inmuno-compromised patients (OR 2.7; 95% CI 1.1-6.7). Resistance to oxacilin was reported in 71.7% of the CoNS isolated. The overall mortality associated to CoNS-B was 6%. Among children with CoNS-PB, 10% received antibiotics, half of them vancomycin. CONCLUSIONS: These results show that CoNS-B occurs mainly as a nosocomial episode. CoNS-PB more likely resulted from specimen contamination at collection, being responsible for almost 60% of all positive blood cultures. The false-positive results caused unnecessary administration of antibiotics in a significant proportion of CoNS-PB events and have a potential impact upon the emergence of resistant pathogens.

Arenavirus nucleocapsid protein displays a transcriptional antitermination activity in vivo.

RNA polymerase pausing and transcriptional antitermination regulates gene activity in several systems. In arenavirus infected cells the switch from transcription to replication is subjected to a hairpin-dependent termination and requires protein synthesis to bypass this signal. The transcriptional antitermination control by Junín virus nucleocapsid protein N, has been demonstrated in vivo by infecting BHK-21 cells expressing this viral protein in the presence of translation inhibitors. This is the first demonstration in vivo of a transcriptional antitermination control in arenavirus-infected cells.

DNA fingerprinting of thermophilic lactic acid bacteria using repetitive sequence-based polymerase chain reaction.

DNA fingerprints of lactic acid bacteria were generated by polymerase chain reaction using a primer based on the repetitive elements found in the genome of Streptococcus pneumoniae (BOX-PCR). The method made it possible to identify 37 isolates from raw milk. industrial starters and yogurt. Differentiation at species, subspecies and strain level was possible for Lactobacillus delbrueckii subsp. lactis, Lb. delbrueckii subsp bulgaricus and Str. thermophilus. BOX-PCR was also applied to studying the strain composition of a starter culture and the direct detection of strains in commercial fermented milk.

[Chemical composition of a typical bread and its energy and protein supply in the population from the northeast of Mexico]. / Composición química del pan tradicional e industrial y su aporte de energía y proteína en la población del Noroeste de México.

Bread from wheat flour is one of the most widely consumed products by the Mexican population. In this study, proximate composition, and mineral content were determined, and cost per/gram of protein and energy of traditional and industrial bread were compared. Seven types of bread were analyzed: bolillo, virginia, white bread and pastries conchita, croissant, breadroll and donuts. Products were analyzed for proximate composition by official methods, and content of Na, K, Ca, Mg, Fe, y Zn by atomic absorption spectrometry. Based on 24 hour recall interviews, the consumption of each type of bread as well as the percent of protein and energy allowances were calculated. Significant differences (p < 0.05) were found with respect to nutrient content between traditional and industrial breads. White bread and pastries from industrial origin showed higher costs (p < 0.05) per gram of protein and energy than traditional products in most cases. Both industrial and traditional breads showed higher fat content than that established by Official Mexican regulations. A high content of Ca was found in bread from the industrial origin and K, Mg, Fe y Zn were higher (p < 0.05) than those from the traditional bakery.

Arenavirus phylogeny: a new insight.

Arenaviridae is a worldwide distributed family, of enveloped, single stranded, RNA viruses. The arenaviruses were divided in two major groups (Old World and New World), based on serological properties and genetic data, as well as the geographic distribution. In this study the phylogenetic relationship among the members of the Arenaviridae was examined, using the reported genomic sequences. The comparison of the aligned nucleotide sequences of the S RNA and the predicted amino acid sequences of the GPC and N proteins, together with the phylogenetic analysis, strongly suggest a possible kinship of Pichindé and Oliveros viruses, with the Old World arenavirus group. This analysis points at the evolutive relationships between the arenaviruses of the Americas and can be used to evaluate the different hypotheses about their origin.

Molecular characterization of attenuated Junin virus strains.

The Junin virus strain Candid #1 was developed as a live attenuated vaccine for Argentine haemorrhagic fever. In this paper we report the nucleotide sequences of S RNA of Candid #1 and its more virulent ancestors XJ#44 and XJ (prototype). Their relationship to Junin virus wild-type MC2 strain and other closely and distantly related arenaviruses was also examined. Comparisons of the nucleotide and amino acid sequences of N and GPC genes from Candid #1 and its progenitor strains revealed some changes that are unique to the vaccine strain. These changes could be provisionally associated with the attenuated phenotype.

Characterization of arenaviruses using a family-specific primer set for RT-PCR amplification and RFLP analysis. Its potential use for detection of uncharacterized arenaviruses.

Arenaviruses are enveloped viruses with a genome composed of two ssRNA species, designated L and S. The arenaviruses were divided in two major groups (Old World and New World), based on serological properties and genetic data, as well as geographic distribution. A sequence alignment analysis of all reported arenavirus S RNAs yielded 17 conserved regions in addition to a reported conserved region at the end of both RNAs. The consensus sequences of these regions were used to design generalized primers suitable for RT-PCR amplification of a set of overlapping nucleotide sequence fragments comprising the complete S RNA of any arenavirus. A restriction analysis (RFLP) was designed to rapidly typify the amplified fragments. This RT-PCR-RFLP approach was tested with Old World (LCM) and New World (Junin and Tacaribe) arenaviruses. Furthermore, using this procedure the whole S RNA of a novel arenavirus isolate obtained from a rodent trapped in central Argentina, was amplified and characterized. Partial nucleotide sequence data were used for phylogenetic analyses that showed the relationships between this arenavirus and the rest of the members of the family. This relatively simple methodology will be useful both in basic studies and epidemiological survey programs.

PCR screening for carriers of bovine leukocyte adhesion deficiency (BLAD) and uridine monophosphate synthase (DUMPS) in Argentine Holstein cattle.

BLAD (Bovine Leukocyte Adhesion Deficiency) and DUMPS (Deficiency of Uridine Monophosphate Synthase) are monogenic autosomal, recessive inherited diseases of Holstein cattle. Single nucleotide changes (point mutations) responsible for the genetic disorders were detected by polymerase chain reaction coupled with restriction fragment length polymorphism assays (PCR-RFLP). Using oligonucleotide primers, DNA fragments of predicted sizes were amplified, and the products' specificity was assessed by nucleotide sequencing. Mutations were detected in DNA samples from bovine blood and semen by the presence or absence of restriction sites within the PCR amplification products (Taq I, Hae III for BLAD, Ava I for DUMPS). The test included 104 bulls and 950 cows of Argentinean Holstein breed. Defective alleles frequencies were as follows: 2.88% BLAD in bulls used in artificial insemination, 1.79% in cows; 0.96% DUMPS in bulls and 0.11% in cows.

Rapid diagnosis of Argentine hemorrhagic fever by reverse transcriptase PCR-based assay.

Argentine hemorrhagic fever (AHF) is an endemo-epidemic disease caused by Junín virus. This report demonstrates that a reverse transcriptase (RT) PCR-based assay developed in our laboratory to detect Junín virus in whole blood samples is sensitive and specific. The experiments were conducted in a double-blinded manner using 94 clinical samples collected in the area in which AHF is endemic. The RT-PCR-based assay was compared with traditional methodologies, including enzyme-linked immunosorbent assay, plaque neutralization tests, and occasionally viral isolation. The calculated parameters for RT-PCR diagnosis, with seroconversion as the "gold standard," were 98% sensitivity and 76% specificity. It is noteworthy that 94% of the patients with putative false-positive results (RT-PCR positive and no seroconversion detected) exhibited febrile syndromes of undefined etiology. These results could be interpreted to mean that most of those patients with febrile syndromes were actually infected with Junín virus but did not develop a detectable immune response. Furthermore, 8 laboratory-fabricated samples and 25 blood samples of patients outside the area in which AHF is endemic tested in a similar way were disclosed correctly (100% match). The RT-PCR assay is the only laboratory test available currently for the early and rapid diagnosis of AHF. It is sensitive enough to detect the low viremia found during the period in which immune plasma therapy can be used effectively, reducing mortality rates from 30% to less than 1%.

A simple nucleic acid amplification assay for the rapid detection of Junín virus in whole blood samples.

Argentine hemorrhagic fever (AHF) is an endemoepidemic disease with cardiovascular, renal and neurologic alterations acquired in the richest farming land in Argentina. It is caused by Junín virus, one of the few human pathogenic arenaviruses. The S RNA of Junín virus has been molecularly cloned and its nucleotide sequence determined in our laboratory. This information was used to develop a rapid nucleic acid-based diagnostic test commensurate with the low viraemia detected in AHF patients. Junín virus-specific cDNA probes labeled using various methods proved insensitive in dot-hybridizations. Therefore, a RT polymerase chain reaction (PCR) was developed using a pair of oligonucleotide primers to reverse-transcribe and amplify the viral S RNA. The amplification of the target sequences was measured by ethidium bromide staining of the DNA fragments after agarose gel electrophoresis. This type of assay allowed the specific detection of Junín virus RNA sequences present in a single infected BHK21 cell over a background of 10(4) uninfected cells. Control reactions were performed on RNA samples extracted from uninfected cells or cells infected with a high multiplicity of LCMV, another arenavirus present in the AHF endemic area. The PCR was first adapted to detect viral RNA in peripheral blood mononuclear cells, described to harbor most of the virus. A simplification of this assay allows the detection of Junín virus in RNA extracted from 100 microliters of whole blood using guanidium thiocyanate disruption and acid phenol extraction. Under the conditions described in this paper, it is possible to detect up to 0.01 pfu of Junín virus in a blood sample. An early and rapid laboratory diagnostic test for AHF is important since the only effective therapy that reduces the mortality rate from 30% to less than 1% consists of early treatment with immune plasma.