Local Variation in Inclusion on Kidney Transplant Waiting Lists: Analysis in the Reference Area of a Kidney Transplant Center.

BACKGROUND: Access for end-stage renal disease (ESRD) patients to the renal transplant (RT) waiting list can vary depending on the criteria used and how they are applied in each dialysis unit. METHODS: This study was performed in the reference area (2.5 million inhabitants) of a transplant center. Data were from a regional registry (Information System of the Autonomous Coordination of Transplants in Andalusia) of total dialysis patients. Patients were grouped according to transplant status as: effective waiting list (WL); never recorded or excluded (E); incomplete immunologic study or discharge data (IIS); temporary contraindication (TC); or active (A). RESULTS: There were 1424 dialysis patients. Of these, 58% were E, 18% were IIS, 14% were TC, and 10% were A. Significant differences were detected for proportion of patients listed as active status (A) in 3 hospital dialysis units (2.9%-13.4%) and 12 hemodialysis centers (4.2%-29.2%); proportion of IIS cases in the hospitals (0%-57%) and dialysis centers (0%-58%); and in proportion of TC cases in the hospitals (19%-50%) and dialysis centers (2.5%-19.3%). The mean age of patients varied significantly between IIS, TC, and A groups (60.3, 54.8, and 52.3 years old, respectively, P < .001). Accentuated differences between the 2 provinces included in the sector were verified. There are notable differences in inclusion of pre-dialysis patients between hospital units. CONCLUSION: We detected considerable variability between hospital units and non-hospital dialysis centers in relation to inclusion on the active transplant waiting list and the proportion of patients with IIS or TC status. It is essential to implement a more homogeneous system for case selection through a specific intervention program from the reference center.

Reasons for Noninclusion on the Kidney Transplant Waiting List: Analysis in a Set of Hemodialysis Centers.

INTRODUCTION: End-stage renal disease patients' access to the renal transplant (RT) waiting list (WL) depends on general criteria and their specific application in the different treatment units. METHODS: Study in nonhospital hemodialysis centers (n = 9), dependent on an adult RT center. Cases included 228 patients considered to have nonactive status on the WL due to incomplete immunologic data (no blood group or HLA typing) or temporary contraindication from an incomplete pretransplant study (nonimmunologic) or comorbidity. Each individual situation was studied by reviewing the center's clinical history with the nephrologist in charge. RESULTS: Three situations were classified three groups. (1) Patients in this group had incomplete basic study (65 patients, 28.5%) pending cardiologic evaluation in 34%; urologic evaluation, 26%; both 18%; others, 9%; study not initiated, 12%. (2) Patients in this group had pre-existing or onset comorbidities (117 patients, 51.3%) pending studies or confirmed resolution: obesity, 30%; cancer, 17%; cardiovascular disease, 14%; digestive pathology, 10%; infection, 9%; neuropsychiatric disorders, 7%; multiple, 13%. (3) Patients in this group had other situations contraindicating RT (46 patients, 20.2%): poor therapeutic adherence, 30%; negative will of the patient, 26%; social issues, 9%; excluded by the center (not reported), 35%. CONCLUSIONS: We detected a high incidence of cases pending basic tests for inclusion on the WL. Obesity can be highlighted as the most frequent cause for noninclusion. Further support and coordination is required with referral hospital centers to increase and refine the RT WL.

Survival and Evolution of Renal Function in Kidney Transplant Recipients From Type II Asystolic Donations: A Single-center Experience.

BACKGROUND: In recent years, stagnation in the number of kidneys from after brain-dead donors (DBD) has stimulated the use of non-heart beating donors (NHBDs). Herein we present our 5-year experience with type II Maastricht NHBDs in renal transplantation. METHODS: All patients (n = 50) in this study received type II Maastricht NHBD kidneys (March 2012 to February 2017), with a median follow-up of 33 months. RESULTS: Mean donor age was 39 ± 12 years, mean creatinine 1.24 ± 0.2 mg/dL, and the most frequently observed blood group (donors and recipients) was type A (64%). Recipients were slightly younger (51 ± 11 years old), with mean time on dialysis of 30 ± 24 months. Almost all were primary transplants. Pre-transplant panel-reactive antibodies (PRA) were <25%; initial immunosuppression was thymoglobulin, corticosteroids, mycophenolate mofetil, and delayed introduction of tacrolimus. Six percent were nonfunctioning kidneys; 79.6% presented with delayed renal function (mean duration 14 ± 9 days). Acute rejection was seen in 6% of patients. Mean creatinine at month 3 was 1.7 ± 0.8 mg/dL, and 1.5 ± 0.8 mg/dL in the first year. The last available mean creatinine was 1.54 ± 0.7 mg/dL. Proteinuria in the third month, first year, and third year was 0.70, 0.41, and 0.26 g/d, respectively. Recipient survival at the first, third, and fifth year was 100%, 100%, and 86%, and when graft-censored for death was 94%, 91%, and 91%, respectively. The incidence of acute rejection during first year was 6%, and 2% in the second year. Exitus incidence was 4% and cytomegalovirus infection was 21.3%. BK viremia between 1000 and 10,000 copies/mL was seen in 4.3%, and reached >10,000 copies/mL in 2.1%. CONCLUSIONS: Type II NHBD has shown limited frequency of nonfunctioning kidney and high functional delay. The results in survival and renal function are very acceptable, comparable with levels seen in donation after brain death.

Resistant Cytomegalovirus Infection After Renal Transplantation: Literature Review.

BACKGROUND: Resistant cytomegalovirus (R-CMV) is an emerging problem in the renal transplantation population. The most frequent CMVs are high-resistance mutations (UL97 gene). METHODS: We describe our experience in management of R-CMV after renal transplant at our center (2012-2016). RESULTS: We encountered 3 cases of R-CMV infection after renal transplant (all primary infections). All 3 patients received induction therapy with corticosteroids, tacrolimus, and mycophenolate mofetil. The first patient (basiliximab induction, preemptive CMV) developed CMV replication on day +53, which responded poorly both to standard-dose valganciclovir (vGCV) and high-dose ganciclovir (GCV) (creatinine clearance [CrCl] >70 mL/min; vGCV 900 mg twice daily for 50 days and GCV 7.5 mg/kg twice daily for 8 days). Hematologic toxicity occurred. The R-CMV test was positive and foscarnet (FOS) was initiated (90 mg/kg twice daily for 21 days). The second patient presented CMV infection (day +30, thymoglobulin induction, CMV prophylaxis), which was not controlled with the high dose (CrCl 23 mL/min; GCV 3.5 mg/kg twice daily and vGCV 900 mg twice daily), resulting in severe neutropenia. R-CMV was detected and FOS initiated (FOS 50 mg/kg twice daily for 7 days and 50 mg/kg every 2 days for 13 days). The third patient's infection occurred on day +22 (basiliximab induction, preemptive CMV). Standard-dose vGCV was uneffective (CrCl >70 mL/min, vGCV 900 mg twice daily) and it did not respond to the high dose (GCV 7.5 mg/kg twice daily and vGCV 2700 mg/d). Moderate hematologic toxicity occurred. R-CMV was diagnosed and FOS treatment begun (FOS 70 mg/kg per day for 2 weeks). CONCLUSIONS: Resistant CMV infection may be severe due to viral infection and side effects of high-dose antiviral treatment. We presented 3 cases requiring the use of FOS in the absence of response or toxic effects from the usual treatment, with an optimal sustained response (temporary in case 2) and without serious side effects.

Hepatitis C Virus in Kidney Transplant Recipients: A Problem on the Path to Eradication.

BACKGROUND: Hepatitis C virus (HCV) still has significant prevalence in kidney transplant (KT) recipients and is related to poor recipient and graft survival. New direct-acting antivirals (DAA) are leading to a radical change in the problem. METHODS: We studied HCV prevalence at the time of transplantation and in follow-up patients, the way cases are handled, and the results of DAA. RESULTS: A total of 2,001 KT had been performed in our center since 1978. Pre- or post-transplantation HCV serology was present in 1,880 cases and was positive in 13.4%. A total of 1,195 transplant recipients were still being monitored by us, with only 60 (5%) HCV+ and 45 (3.6%) RNA+ cases. Of these 45 HCV+/RNA+, 25 had been or were being treated, 7 were about to begin treatment, 1 was awaiting new DAA treatment owing to low glomerular filtration rate (GFR), 3 were being evaluated, 2 had been excluded owing to high comorbidity, 2 refused to be treated, 2 needed to return to hemodialysis, and 1 was lost to follow-up. Except 1 case where Viekira Pak was used because of low GFR, all cases included sofosbuvir as the main drug associated with either ledipasvir (70%) or daclatasvir (25%). Ribavirin was added as coadjuvant in 35% of cases. Twenty-one patients had completed treatment (84%). Two patients had to interrupt DAA therapy (8%), one because of hepatotoxicity and the other as a result of a liver transplantation. In every case, the graft maintained function and negativization of viral replication occurred. CONCLUSIONS: Side effects have been low, anemia related to ribavirin being the main one. Just one case needed to be interrupted at the 7th week of DAA therapy due to hepatotoxicity. It has frequently been necessary to adjust immunosuppression treatment with the use of higher doses of tacrolimus.

Assessment of the Renal Function in Potential Donors of Living Kidney Transplants: Expanded Study.

INTRODUCTION: It is very important to determine as accurately as possible the renal function in potential living renal transplant donors, especially those with limited renal function (CrCl <90 mL/m/1.73 m(2)), age older than 50 years, and cardiovascular risk factors that might favor the development of long-term kidney diseases. OBJECTIVE: The objective of this study was to compare the direct measurement of glomerular filtration rate (GFR) using EDTA-Cr51 and the estimations based on creatinine (eGFR): Cr clearance (CCr) with 24-hour urine and estimated using Cockroft-Gault (adjusted by using body surface area-Mosteller formula-SC), MDRD-4, MDRD-6, and CKD-EPI to determine the usefulness of different methods from EDTA-Cr51to evaluate the kidney function. PATIENTS AND METHODS: The kidney function evaluation has been made to 105 potential kidney donors using the EDTA-Cr51 method. The GFR obtained through the EDTA-Cr51 is compared with the CCr values in 24-hour urine and eGFR based on creatinine (Cockcroft-Gault, MDRD4, MDRD6, and CKD-EPI). RESULTS: Using the Bland Altman graphic we have observed that the most dispersed results are obtained with the eGFR using CCr in 24-hour urine and CKD-EPI. By means of Pasing & Bablock, we realized that MDRD-4 and MDRD-6 show the highest approximation to the reference method proposed to be substituted, whereas CCr shows a high dispersion. CONCLUSIONS: eGFR using MDRD-4 and MDRD-6 formulas reveal the best adjustment to the measure by EDTA-Cr51. This might represent the best option if a direct eGFR measure is not available.

New Antivirals for Hepatitis C Infection Among Infected Kidney Transplant Recipients: A Case Report.

The most common hepatopathy in end-stage renal disease is chronic hepatitis C virus (HCV) infection, which decreases allograft and patient survival in kidney transplants. Until last year we did not have treatments free of interferon, which was contraindicated after renal transplantation owing to the risk of allograft rejection. Recently, new drugs have been discovered for interferon-free regimens. These drugs present a cure rate of up to 90% and can be used in transplant recipients. Here we present our 1st 3 cases. In our experience, new antivirals have proven to be effective and safe for the treatment of HCV hepatopathy in kidney transplant recipients and liver-kidney transplantation, thus helping us to prevent complications related to HCV infection in transplant recipients.

Study of renal function in living kidney donors: estimated or measured glomerular filtration.

INTRODUCTION: In living kidney donations the accuracy of renal function is fundamental, especially for potential donors who have limited renal function (creatinine clearance levels [CCr] <90 mL/m/1.73 m(2)), are >50 years old, and who have cardiovascular risk factors that might favor the development of kidney diseases. OBJECTIVE: To compare the direct measured glomerular filtration (mGFR) using 51Cr-EDTA and the estimations based on creatinine (estimated glomerular filtration rate [eGFR]): CCr with 24-hour urine, and estimated using Cockroft-Gault (adjusted using body surface area, Mosteller formula), modification of diet in renal disease-4 (MDRD-4), MDRD-6, and Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) to determine the usefulness of different methods to evaluate the kidney function. PATIENTS AND METHODS: The kidney function evaluation was performed for 37 potential kidney donors using the 51Cr-EDTA method. The GFR obtained through the 51Cr-EDTA was compared with the CCr values in 24-hour urine and eGFR based on creatinine (Cockcroft-Gault, MDRD-4, MDRD-6, and CKD-EPI). RESULTS: Using the Bland Altman graph, the most dispersed results were obtained with the eGFR using CCr in 24-hour urine and CKD-EPI. By means of Passing and Bablok, MDRD-4 and MDRD-6 showed the highest approximation to the reference method proposed to be substituted, whereas CCr showed a high dispersion. CONCLUSION: The eGFR using MDRD-4 and MDRD-6 formulas revealed the best adjustment to the measure by 51Cr-EDTA. This might represent the best option if a direct eGFR measure is not available.

Effect of barrel design and the inoculation of Acetobacter pasteurianus in wine vinegar production.

The traditional production of wine vinegar is a lengthy process with little or no microbiological control. The aim of this study was to shorten the acetification process via three different strategies: changes in wood type; barrel shape; and the inoculation of an Acetobacter pasteurianus pure culture. The barrel shape was modified by constructing two prototypes with higher liquid-air interface. We compared the changes in acetic acid bacteria (AAB) population dynamics in these barrels with those of a submerged method. The wood type had no effect on the acetification length, whereas the shape of the barrel resulted in a significant shortening of the acetification length. Although the selected AAB strain did not always take over, it reduced the biodiversity of the AAB. The inoculated strain was predominant in oak barrels, whereas in the highly aerated prototypes Gluconacetobacter species (Ga. intermedius and/or Ga. europaeus) displaced A. pasteurianus, as what occurs in the submerged method.

Quantification of the expression of reference and alcohol dehydrogenase genes of some acetic acid bacteria in different growth conditions.

AIMS: The aim of this study was to develop a reliable system to analyse the expression of the pyrroloquinoline quinone (PQQ)-alcohol dehydrogenase (ADH) and test its ability to predict the growth and oxidative activity of some acetic acid bacteria (AAB). METHODS AND RESULTS: Specific primers were designed for use in RT-PCR to quantify ADH expression and several housekeeping genes in four species of AAB. 16S rRNA gene was selected as an internal control. The relative expression of adhA was measured in Acetobacter aceti, Acetobacter pasteurianus, Gluconacetobacter hansenii and Gluconobacter oxydans grown in two media that had glucose or ethanol as the carbon source. AAB adhA expression was shown to be related to the two Acetobacter species' ability to oxidise and grow on ethanol, whereas G. oxydans were unable to grow on ethanol and the growth of Ga. hansenii was not related to adhA expression. CONCLUSIONS: The differential expression of ADH could be a marker to analyse both growth and oxidation ability in some AAB, especially those of the genus Acetobacter. SIGNIFICANCE AND IMPACT OF THE STUDY: Several housekeeping genes were tested in AAB and after growth in different media and it was evident that only the ribosomal coding genes were adequate as reference genes for RT-PCR.

Enfermedad por depósito de cadenas ligeras. Experiencia en nuestro medio / No disponible

La enfermedad por depósito de cadenas ligeras (EDCL) es una entidad rara, caracterizada por el depósito de un solo tipo de cadena ligera en la membrana basal del riñón. Puede asociarse a una discrasia de células plasmáticas, aunque en ocasiones no se detecta patología hematológica y se denomina idiopática. Suele manifestarse como una insuficiencia renal severa con proteinuria nefrótica, no tiene tratamiento claramente establecido y el pronóstico es malo. El objetivo de este trabajo es analizar las características de los casos de EDCL diagnosticados en nuestro medio. Se identifican 6 casos, todos entre 1999 y 2005 de un total de 640 biopsias realizadas en ese periodo, 4 mujeres y 2 varones, media de 57 años. Se detectó un mieloma en 3 pacientes(50%). La insuficiencia renal aguda o de rápida evolución fue la presentación clínica más frecuente (66%) junto con proteinuria nefrótica (66%). Todas las biopsias mostraban engrosamiento de la membrana basal tubular y depósito lineal de cadenas kappa en la misma. La lesión glomerular más frecuente fue la glomérulo esclerosis nodular (83%).En un caso la afectación fue exclusivamente túbulo intersticial con cilindros tubulares asociados. Se trataron 3 pacientes,2 con mieloma. Requirieron diálisis 5 pacientes: 3 con EDCL idiopática con un tiempo medio desde el diagnóstico hasta recibir la misma de 7 días, y 2 con mieloma que tardaron una media de 46 días en requerir diálisis. Fallecieron 4 pacientes, 2 con mieloma. El tiempo de seguimiento hasta el exitus fue de 13 semanas para los pacientes con mieloma y de 110 semanas para el resto. Conclusión, la EDCL parece mas frecuente de lo publicado y se asocia a mieloma en la mitad de los casos. Se presenta con daño renal severo y la evolución renal y del paciente es mala (AU)

The Light chain deposition disease (LCDD) is a strange entity characterized by the deposition of only one type of light chain in the renal tubular basement membranes. It can be associated to a plasmacell dyscrasia, however, it can occur in the absence of any detectable hematological disorder and it is called idiopathic LCDD. The clinical manifestation is renal insufficiency and nephrotic proteinuria, it does not have a clearly fixed treatment and has a severe prognosis. The aim of this work is to analyse the characteristics of the LCDD cases diagnosed within our environment. Six cases were identified, all of them between 1999 and 2005,from a total amount of 640 renal biopsies performed during this period, 4 women and 2 men, average age of 57. Multiple myelomain 3 patients was detected (50%). The acute renal failure or rapidly progressive renal insufficiency was the most frequent clinical presentation (66%) together with nephrotic proteinuria (66%). All the biopsies showed tubular basement membranes thickening and kappa chains with a linear distribution within the same. The most frequent glomerular pathological finding was the nodular sclerosing glomerulopathy (83%). In one of the cases the affectation was exclusively tubular interstitial with tubular casts. 3 patients were treated, 2 with multiple myeloma. 5 patients needed dialysis: 3 with idiopathic LCDD within an average time of 7 days from the diagnosis to its reception, and 2 with myeloma, who started needing dialysis in an average of 46 days. 4 patients died, 2 of them with myeloma. The monitoring time until the death was 13 weeks for the patients with myeloma and 110 weeks for the rest. Conclusion: The LCDD seems to be more frequent than what has been published and it is associated to the myeloma in half of the cases. It appears together with severe renal insufficiency and the patient's and renal prognosis is poor (AU)

[Light chain deposition disease. Experience in our environment]. / Enfermedad por depósito de cadenas ligeras. Experiencia en nuestro medio.

UNLABELLED: The Light chain deposition disease (LCDD) is a strange entity characterised by the deposition of only one type of light chain in the renal tubular basement membranes. It can be associated to a plasma cell dyscrasia, however, it can occur in the absence of any detectable hematological disorder and it is called idiopathic LCDD. The clinical manifestation is renal insufficiency and nephrotic proteinuria, it does not have a clearly fixed treatment and has a severe prognosis. The aim of this work is to analyse the characteristics of the LCDD cases diagnosed within our environment. Six cases were identified, all of them between 1999 and 2005, from a total amount of 640 renal biopsies performed during this period, 4 women and 2 men, average age of 57. Multiple myeloma in 3 patients were detected (50%). The acute renal failure or rapidly progressive renal insufficiency was the most frequent clinical presentation (66%) together with nephrotic proteinuria (66%). All the biopsies showed tubular basement membranes thickening and kappa chains with a linear distribution within the same. The most frequent glomerular pathological finding was the nodular sclerosing glomerulopathy (83%). In one of the cases the affectation was exclusively tubular interstitial with tubular casts. 3 patients were treated, 2 with multiple myeloma. 5 patients needed dialysis: 3 with idiopathic LCDD within an average time of 7 days from the diagnosis to its reception and 2 with myeloma, who started needing dialysis in an average of 46 days. 4 patients died, 2 of them with myeloma. The monitoring time until the death was 13 weeks for the patients with myeloma and 110 weeks for the rest. CONCLUSION: The LCDD seems to be more frequent than what has been published and it is associated to the myeloma in half of the cases. It appears together with severe renal insufficiency and the patient's and renal prognosis is poor.

Application of molecular methods for the differentiation of acetic acid bacteria in a red wine fermentation.

AIMS: To apply rapid and reliable molecular techniques for typing acetic acid bacteria and studying their population dynamics during wine-making processes. METHODS AND RESULTS: We tested the usefulness of the Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) and Repetitive Extragenic Palindromic-PCR (REP-PCR) techniques with reference strains of most of the species of acetic acid bacteria. We obtained exclusive patterns for each strain with the ERIC-PCR technique, proving the utility for characterizing below species level. REP-PCR technique was not as adequate for this purpose because some strains yielded identical fingerprint. One hundred twenty isolates from a commercial red wine fermentation were fingerprinted using both techniques. We detected a high degree of strain diversity in the first stage of fermentation that decreased throughout the process. However, several strains and species were dominant in the alcoholic fermentation phases. The identification of different strains or genotypes at the species level was carried out by restriction analysis of the 16S ribosomal DNA gene. Gluconobacter oxydans dominated the fresh must, while Acetobacter aceti was the only isolated species at the end of the process. Gluconacetobacter hansenii and G. liquefaciens were also isolated in significant numbers at the beginning of fermentation. CONCLUSIONS: ERIC-PCR and REP-PCR techniques proved useful for characterizing strains of acetic acid bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The availability of molecular techniques for a fast and reliable genotypic characterization should increase our knowledge of the ecology of acetic acid bacteria and determine more accurately their growth behaviour during various stages of vinification.

[Hospitalizations preventable by timely and effective primary health care]. / Hospitalizaciones prevenibles mediante una atención primaria oportuna y efectiva.

OBJECTIVE: To know the specific health problems referred as ambulatory care sensitive conditions (ACSC) and to identify the primordial interventions of primary health care (PHC) in reducing hospitalisations due to ACSC. DESIGN: Descriptive study of hospital discharges generated in Catalonia during 1998-1999, and a Delphi study to reach information about PHC primordial interventions. MEASURES: Diagnostic codes of ACSC selected as markers of PHC effectiveness were used. We analysed hospital discharge distribution by age groups and overall, and hospitalisation rates with its 95% confidence intervals. Descriptive analysis of consensus reached by experts using self-administrated questionnaires was done. RESULTS: The 8.42% of total discharges were due to ACSC. The majority of these (86.9%) fell in 4 of the 13 diagnostic categories included in the ACSC list. A great variety of pathologies with different frequencies were identified. Primary prevention and early diagnoses and treatment were considered as primordial interventions. Chronic health problems needed multimodal interventions. CONCLUSIONS: Diagnostic codes included in each diagnostic category were congruent with the diseases identified. Interventions that could prevent hospitalisations due to ACSC are contemplated as role of PHC. Indicator validity to assess PHC effectiveness is maintained by both results.

Hospitalizaciones prevenibles mediante una atención primaria oportuna y efectiva / Hospitalisations preventable by timely and effective primary health care

Objetivo. Conocer los problemas de salud específicos a los que hacen referencia los ambulatory care sensitive conditions (ACSC) e identificar las intervenciones prioritarias de la atención primaria de salud (APS) que podrían reducir las hospitalizaciones por ACSC. Diseño. Estudio descriptivo de las altas hospitalarias generadas por la población de Cataluña durante 1998-1999, y estudio Delphi para obtener la información sobre las intervenciones de la APS. Mediciones principales. Se ha utilizado el listado de códigos de diagnóstico de ACSC considerados marcadores para evaluar la efectividad de la APS. Análisis de la distribución de las altas por grupo de edad y total, y cálculo de las tasas de hospitalización y sus correspondientes intervalos de confianza del 95 per cent. Análisis descriptivo del consenso obtenido de un grupo de expertos mediante cuestionarios autoadministrados. Resultados. El 8,42 per cent del total de altas registradas se considera ACSC. De ellas, un 86,9 per cent corresponde a 4 categorías diagnósticas de las 13 que conforman el listado de ACSC. Se identifica una gran variedad de patologías con frecuencias muy dispares. La prevención primaria y el diagnóstico precoz y tratamiento constituyen las actividades prioritarias. La patología crónica requiere intervenciones multimodales. Conclusiones. Los códigos de diagnóstico incluidos en cada categoría diagnóstica son coherentes con la patología que identifican. Las intervenciones que podrían prevenir las hospitalizaciones por ACSC se consideran hegemónicas de la APS. Estos resultados sustentan la validez del indicador para medir la efectividad de la APS (AU)

Yeast population dynamics in spontaneous fermentations: comparison between two different wine-producing areas over a period of three years.

Yeast ecology, biogeography and biodiversity are important and interesting topics of research. The population dynamics of yeasts in several cellars of two Spanish wine-producing regions was analysed for three consecutive years (1996 to 1998). No yeast starter cultures had been used in these wineries which therefore provided an ideal winemaking environment to investigate the dynamics of grape-related indigenous yeast populations. Non-Saccharomyces yeast species were identified by RFLPs of their rDNA, while Saccharomyces species and strains were identified by RFLPs of their mtDNA. This study confirmed the findings of other reports that non-Saccharomyces species were limited to the early stages of fermentation whilst Saccharomyces dominated towards the end of the alcoholic fermentation. However, significant differences were found with previous studies, such as the survival of non-Saccharomyces species in stages with high alcohol content and a large variability of Saccharomyces strains (a total of 112, all of them identified as Saccharomyces cerevisiae) with no clear predominance of any strain throughout all the fermentation, probably related to the absence of killer phenotype and lack of previous inoculation with commercial strains.

Identification of acetic acid bacteria by restriction fragment length polymorphism analysis of a PCR-amplified fragment of the gene coding for 16S rRNA.

Acetic acid bacteria (AAB) irreversibly spoil wines and represent a serious problem. Limited studies on the ecology of AAB during winemaking have been done due to the lack of rapid and precise techniques for their identification. RFLP analysis of PCR-amplified fragment of 16S rDNA was performed on AAB reference strains. The amplified rDNAs were approximately 870-bp long for all AAB species while no amplicons were detected for lactic acid bacteria and yeasts. Out of the four restriction enzymes tested, TaqI was the most efficient one and divided the studied AAB into six groups. However, complete differentiation among collection strains of Acetobacter pasteurianus and Gluconoacetobacter hansenii was not possible.

Are patients with hyperlipidemia being treated? Investigation of cholesterol treatment practices in an HMO primary care setting.

BACKGROUND: The main study objective was to investigate cholesterol treatment practices of primary care physicians in a managed care setting. METHODS: The study was a retrospective review of data with a quasiexperimental design. The National Cholesterol Education Program-Adult Treatment Panel II (NCEP-ATP II) guidelines were used as the reference for conducting a measurement model in the study. Data were randomly selected via a systematic probability sampling method from a health maintenance organization (HMO) capitated risk-based contracting medical clinic in southern Florida. RESULTS: Of the 348 patients selected for the study, 224 (65%) needed either dietary therapy (n = 106) or drug therapy (n = 118). However, only 16 patients (13.6%) had ever had cholesterol-lowering drug regimens prescribed during the 5-year study period. CONCLUSIONS: Our findings indicate that (1) primary care physicians have poorly adopted the cholesterol management practice recommended by NCEP guidelines and need to improve their recognition and treatment of hypercholesterolemia; and (2) the problem of underutilizing prescription medications may be associated with risk-sharing capitation arrangements between physicians and third-party insurers.

Identification of acetic acid bacteria by RFLP of PCR-amplified 16S rDNA and 16S-23S rDNA intergenic spacer.

DNA corresponding to 16S rDNA and the 165-23S rDNA intergenic spacer (ITS) from 22 reference strains of acetic acid bacteria, representing the diversity of the family Acetobacteraceae, and 24 indigenous acetic acid bacteria isolated from wine fermentations were analysed by PCR-RFLP. Frateuria aurantia LMG 1558T and Escherichia coli ATCC 11775T were included as outgroups. PCR-amplified products of about 1450 bp were obtained from the 16S rDNA of all the strains and products of between 675 and 800 bp were obtained from the 16S-23S rDNA ITS. PCR products were digested with 4-base-cutting restriction enzymes in order to evaluate the degree of polymorphism existing among these strains. Of the enzymes tested, Taql and Rsal were the most discriminatory and showed no intraspecific variations in the restriction patterns. Restriction analysis of the 16S rDNA with these enzymes is proposed as a rapid and reliable method to identify acetic acid bacteria at the level of genus and species (or related species group) and its applicability to identification of indigenous acetic acid bacteria was demonstrated. The same degree of distinction as that for the 16S rDNA analysis was obtained within reference strains of acetic acid bacteria by PCR-RFLP of the 16S-23S rDNA ITS. However, 16S-23S rDNA ITS restriction patterns of strains isolated from wine did not match those of any of the reference strains. Thus, PCR-RFLP of the 16S-23S rDNA ITS is not a useful method to identify isolates of acetic acid bacteria at the species level, although it may be an adequate method to detect intraspecific differentiation.

Molecular analysis of yeast population dynamics: effect of sulphur dioxide and inoculum on must fermentation.

The effects of sulphur dioxide and a commercial starter inoculum on yeast population dynamics have been analysed by a molecular approach. Yeast identification from fermenting Carinyena grape musts was performed by RFLP's of mtDNA and rRNA-coding DNA. As expected, the use of a commercial inoculum speeded up the start of fermentation, while SO2 addition limited the development of non-Saccharomyces species. However, this effect was also observed with yeast inoculation. Further analysis of population dynamics could lead to a recommendation for the reduction of the dosage of SO2 by the addition of appropriate inoculum of yeasts in the must. Furthermore, the timing of inoculum addition could be modified to allow a proper contribution of non-Saccharomyces species. Molecular biology analysis of population dynamics could provide a tool to efficiently reduce the dosage of SO2 and adjust the timing of inoculum addition.