DNA Extraction Methods to Obtain High DNA Quality from Different Plant Tissues.

DNA extraction from plant samples is very important for a good performance of diagnostic molecular assays in phytopathology. The variety of matrices (such as leaves, roots, and twigs) requires a differentiated approach to DNA extraction. Here we describe three categories of matrices: (a) symptomatic bark/wood tissue; (b) residues of frass resulting from insect woody trophic activities, portions of the galleries produced in the wood, and tissues surrounding exit holes; and (c) leaves of different plant species. To improve the performances of diagnostic assays, we here describe DNA extraction procedures that have been optimized for each matrix type.

Loop-Mediated Isothermal Amplification (LAMP) and SYBR Green qPCR for Fast and Reliable Detection of Geosmithia morbida (Kolarik) in Infected Walnut.

Walnut species (Juglans spp.) are multipurpose trees, widely employed in plantation forestry for high-quality timber and nut production, as well as in urban greening as ornamental plants. These species are currently threatened by the thousand cankers disease (TCD) complex, an insect-fungus association which involves the ascomycete Geosmithia morbida (GM) and its vector, the bark beetle Pityophthorus juglandis. While TCD has been studied extensively where it originated in North America, little research has been carried out in Europe, where it was more recently introduced. A key step in research to cope with this new phytosanitary emergency is the development of effective molecular detection tools. In this work, we report two accurate molecular methods for the diagnosis of GM, based on LAMP (real-time and visual) and SYBR Green qPCR, which are complimentary to and integrated with similar recently developed assays. Our protocols detected GM DNA from pure mycelium and from infected woody tissue with high accuracy, sensitivity, and specificity, without cross-reactivity to a large panel of taxonomically related species. The precision and robustness of our tests guarantee high diagnostic standards and could be used to support field diagnostic end-users in TCD monitoring and surveillance campaigns.

Long-Term Effects on Preventing Frailty and Health Care Costs Associated with a Multifactorial Intervention in the Elderly: Three-Year Follow-Up Data from the Pre-Frail 80 Study.

INTRODUCTION: Preventing or delaying frailty has important benefits. Studies show the effectiveness of multifactorial interventions in the frail and pre-frail elderly, but few have evaluated their long-term effectiveness. Frailty and its consequences have been shown to increase the use of health resources. The main aim was to evaluate the long-term effect of a multifactorial primary healthcare intervention in pre-frail elderly people at 36 months and determine the health resources used and their cost. METHODS: A follow-up of a cohort study of patients who participated in a randomized clinical trial in an urban primary care centre in Barcelona was carried out. We included 200 non-institutionalized people aged ≥80 years who met the Fried pre-frailty criteria. The intervention group (IG) received a 6-month interdisciplinary intervention based on physical exercise, Mediterranean diet advice, assessment of inadequate prescribing in polypharmacy patients, and social assessment, while the control group (CG) received standard of care primary healthcare treatment. Sociodemographic variables were collected at baseline. The Fried criteria, comorbidities, and geriatric syndromes were collected at baseline and 12 and 36 months. For the analysis of health costs, data were collected on visits, complementary tests, hospital admissions, and surgical interventions in the last 36 months. Complexity, the rate of expected emergency admission, and the rate of expected mortality were collected at 36 months. Between-group characteristics were compared at baseline and 36 months using the χ2 test and the t test for independent samples. The post-intervention (12-month follow-up) versus longitudinal follow-up (36-month follow-up) comparison used McNemar's test for each group. The nonparametric Mann-Whitney test was used to compare health costs. RESULTS: Of the 200 patients initially included, we evaluated 135 (67.5%) patients who completed the 36-month follow-up. The mean age was 88.5 years and 64.4% were female. At 36 months, the transition to frailty was much lower in the IG than in the CG (22.1% vs. 32.8%, p = 0.013). The total mean health cost at 36 months was 3,110 EUR in the CG and 2,679 EUR in the IG. No significant between-group differences were observed according to Clinical Risk Groups. CONCLUSIONS: A multifactorial, interdisciplinary intervention carried out in primary care prevented frailty in pre-frail elderly people at 36-month follow-up. Although the IG was grouped into higher grade Clinical Risk Groups and therefore had greater morbidity, the cost was lower than that in the CG.

A therapeutic education program with a diabetes specialist nurse for type 2 diabetes patients using insulin in a primary care setting. A diabetes education program with a diabetes specialist nurse in a primary care setting.

INTRODUCTION: Therapeutic education is an essential part in the management of type 2 diabetes mellitus (T2D). Implementing a therapeutic education program with the participation of a diabetes specialist nurse (DSN) addressed to patients with T2D using more than 2 insulin injections and sub-optimal metabolic control in primary care (PC) could improve health care and clinical outcomes. Our purpose was to evaluate the clinical, educational and patient satisfaction outcomes of this program. MATERIAL AND METHODS: A prospective, longitudinal study was performed with an evaluation before and after the intervention. The program had a duration of 6 months and included individual on-site, phone and group visits. RESULTS: 184 subjects were included and 161 were finally evaluated. 89.4% were included due to sub-optimal metabolic control and 10.6% because of repeated hypoglycemia. In the first group, the mean reduction in HbA1c was -1.34%±1.45% without any increase in hypoglycemia episodes. In the second group, a significant reduction in hypoglycemia episodes/week was observed (2.52±1.66 vs. 0.53±1.06; p<0.05) without any increase in HbA1c. Learning skills, lifestyle, adherence to care, and the perception of quality of life had significantly improved at 6 months (p<0.05). The overall program was positively evaluated by patients, the role of DSN being considered essential by 98% of the responders. CONCLUSION: A structured therapeutic education program, including a DSN, addressed to insulin treated T2D patients attending primary care facilities and with sub-optimal metabolic control is associated with beneficial effects in terms of clinical, educational and patient satisfaction endpoints.

A Therapeutic Education Program with a Diabetes Specialist Nurse for Type 2 Diabetes Patients Using Insulin in a Primary Care Setting. A Diabetes Education Program with a Diabetes Specialist Nurse in a Primary Care Setting. / Resultados del programa de educación terapéutica de optimización dirigido a pacientes insulinizados con diabetes tipo 2 desarrollado por enfermería de práctica avanzada en diabetes en el ámbito de atención primaria.

INTRODUCTION: Therapeutic education is an essential part in the management of type 2 diabetes mellitus (T2D). Implementing a therapeutic education program with the participation of a diabetes specialist nurse (DSN) addressed to patients with T2D using more than 2insulin injections and sub-optimal metabolic control in primary care (PC) could improve health care and clinical outcomes. Our purpose was to evaluate the clinical, educational and patient satisfaction outcomes of this program. MATERIAL AND METHODS: A prospective, longitudinal study was performed with an evaluation before and after the intervention. The program had a duration of 6 months and included individual on-site, phone and group visits. RESULTS: 184 subjects were included and 161 were finally evaluated. 89.4% were included due to sub-optimal metabolic control and 10.6% because of repeated hypoglycemia. In the first group, the mean reduction in HbA1c was -1.34±1.45% without any increase in hypoglycemia episodes. In the second group, a significant reduction in hypoglycemia episodes/week was observed (2.52±1.66 vs. 0.53±1.06; P<.05) without any increase in HbA1c. Learning skills, lifestyle, adherence to care, and the perception of quality of life had significantly improved at 6 months (P<.05). The overall program was positively evaluated by patients, the role of DSN being considered essential by 98% of the responders. CONCLUSION: A structured therapeutic education program, including a DSN, addressed to insulin treated T2D patients attending primary care facilities and with sub-optimal metabolic control is associated with beneficial effects in terms of clinical, educational and patient satisfaction endpoints.

[Effectiveness of an educational program for respiratory rehabilitation of Chronic Obstructive Pulmonary Disease patients in Primary Care in improving the quality of life, symptoms, and clinical risk]. / Efectividad de un programa educativo de rehabilitación respiratoria en atención primaria para mejorar la calidad de vida, la sintomatología y el riesgo clínico de los pacientes con enfermedad pulmonar obstructiva crónica.

AIM: To determine the impact of an educational program to improve the management of chronic obstructive pulmonary disease (COPD) that contributes to an increase of the quality of life, exercise capacity, level of dyspnoea, and clinical risk. DESIGN: Intervention study without controls. LOCATION: Primary Healthcare Centre. PARTICIPANTS: 193 patients with COPD were invited, 73 accepted and 55 participated in the educational program. INTERVENTIONS: Respiratory rehabilitation educational program with basic concepts of pulmonary and respiratory pathophysiology, respiratory physiotherapy exercises, practical workshop on the use of the most frequent inhalation devices, understanding of chronic disease and self-care measures in case of exacerbation. MAIN MEASUREMENTS: The quality of life (the COPD assessment test), exercise tolerance (the Six-Minute Walk Test), rating of perceived exertion (Borg Dyspnoea Score) and clinical risk (BODE index) were assessed by means of validated questionnaires in Spanish. RESULTS: A total of 43 (78.2%) participants completed the program. An improvement in the quality of life by a mean of 3.3 points was observed (95%CI; 1.76-4.84). Just over half (53.5%) of the participants obtained a clinically relevant improvement. Participants also improved their physical exercise capacity at post-intervention by increasing the distance that they walked in 6min by a mean of 20.76m (95%CI; 2.57-38.95). Improvements in the level of dyspnoea and clinical risk were also observed. CONCLUSIONS: The educational program shows a statistically significant and clinically relevant improvement in the quality of life, fatigue, symptomatology, exercise capacity, level of dyspnoea, and clinical risk. The program is adaptable to the health care routine of healthcare centres.