A new multiplex PCR protocol to detect mixed trypanosomatid infections in species of Apis and Bombus.

Trypanosomatids are highly prevalent pathogens of Hymenoptera; however, most molecular methods used to detect them in Apis and Bombus spp. do not allow the identification of the infecting species, which then becomes expensive and time consuming. To overcome this drawback, we developed a multiplex PCR protocol to readily identify in a single reaction the main trypanosomatids present in these hymenopterans (Lotmaria passim, Crithidia mellificae and Crithidia bombi), which will facilitate the study of their epidemiology and transmission dynamics. A battery of primers, designed to simultaneously amplify fragments of the RNA polymerase II large subunit (RPB1) of L. passim, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of C. mellificae and the DNA topoisomerase II (TOPII) of C. bombi, was tested for target specificity under single and mixed template conditions using DNA extracted from cell cultures (L. passim ATCC PRA403; C. mellificae ATCC 30254) and from a bumblebee specimen infected with C. bombi only (14_349). Once validated, the performance of the method was assessed using DNA extractions from seven Apis mellifera (Linnaeus, 1758) and five Bombus terrestris (Linnaeus, 1758) field samples infected with trypanosomatids whose identity had been previously determined by PCR-cloning and sequencing (P-C-S). The new method confirmed the results obtained by P-C-S: two of the honeybee samples were parasitized by L. passim, C. mellificae and C. bombi at the same time, whereas the other five were infected with L. passim only. The method confirmed the simultaneous presence of L. passim and C. mellificae in two B. terrestris, where these parasites had not previously been reported.

Course of COPD assessment test (CAT) and clinical COPD questionnaire (CCQ) scores during recovery from exacerbations of chronic obstructive pulmonary disease.

INTRODUCTION: COPD exacerbations have a negative impact on lung function, decrease quality of life (QoL) and increase the risk of death. The objective of this study was to assess the course of health status after an outpatient or inpatient exacerbation in patients with COPD. METHODS: This is an epidemiological, prospective, multicentre study that was conducted in 79 hospitals and primary care centres in Spain. Four hundred seventy-six COPD patients completed COPD assessment test (CAT) and Clinical COPD Questionnaire (CCQ) questionnaires during the 24 hours after presenting at hospital or primary care centres with symptoms of an exacerbation, and also at weeks 4-6. The scores from the CAT and CCQ were evaluated and compared at baseline and after recovery from the exacerbation. RESULTS: A total of 164 outpatients (33.7%) and 322 inpatients (66.3%) were included in the study. The majority were men (88.2%), the mean age was 69.4 years (SD = 9.5) and the mean FEV1 (%) was 47.7% (17.4%). During the exacerbation, patients presented high scores in the CAT: [mean: 22.0 (SD = 7.0)] and the CCQ: [mean: 4.4 (SD = 1.2)]. After recovery there was a significant reduction in the scores of both questionnaires [CAT: mean: -9.9 (SD = 5.1) and CCQ: mean: -3.1 (SD = 1.1)]. Both questionnaires showed a strong correlation during and after the exacerbation and the best predictor of the magnitude of improvement in the scores was the severity of each score at onset. CONCLUSIONS: Due to their good correlation, CAT and CCQ can be useful tools to measure health status during an exacerbation and to evaluate recovery. However, new studies are necessary in order to identify which factors are influencing the course of the recovery of health status after a COPD exacerbation.

Alopecia Areata in Latin America: Where are We and Where are We Going?

Alopecia areata (AA) represents an underrecognized burden in Latin America (LA), severely impacting quality of life (QoL). This impact is exacerbated by limited access to specialized dermatologic care and therapies for AA within and among nations. Many of the unmet needs for AA globally also exist in LA. The region has geographic, ethnic, cultural, and economic conditions. With new AA medicines targeting immunologic pathways on the horizon, LA must prepare regarding regulatory issues, reimbursement, awareness, and education to give adequate and timely treatment for patients with AA. To address these issues, the Americas Health Foundation convened a panel of six dermatologists from Argentina, Brazil, Colombia, and Mexico who are experts in AA and its comorbidities for a 3-day virtual meeting to discuss AA diagnosis and treatment in LA and create a manuscript offering recommendations to address discussed barriers. This publication examines unmet AA needs in LA, treatment, and innovative therapies and recommends improving AA care. Access constraints to conventional and novel medicines hinder appropriate treatments for patients. Therapy initiation delays can affect QoL, mental health, and disease progression. People with AA face stigmas, discrimination, and misconceptions owing to a lack of disease awareness. With promising new treatments for AA on the horizon, all stakeholders must coordinate efforts to enhance LA's AA management landscape and improve patient outcomes.

Calcium enhances polyhydroxyalkanoate production and promotes selective growth of the polyhydroxyalkanoate-storing biomass in municipal activated sludge.

Activated sludge from municipal wastewater treatment processes can be used directly for the production of biodegradable polyesters from the family of polyhydroxyalkanoates (PHAs). However, municipal activated sludge typically cannot accumulate PHAs to very high levels and often low yields of polymer produced on substrate are observed. In the present work, it was found that the presence of calcium promotes selective growth and enrichment of the PHA-storing biomass fraction and significantly improved both PHA contents and yields. Calcium addition resulted in PHA contents of 0.60 ± 0.03 gPHA/gVSS and average PHA yields on substrate of 0.49 ± 0.03 gCODPHA/gCODHAc compared to 0.35 ± 0.01 gPHA/gVSS and 0.19 ± 0.01 gCODPHA/gCODHAc without calcium addition. After 48 h, three times more PHA was produced compared to control experiments without calcium addition. Higher PHA content and selective biomass production is proposed to be a consequence of calcium dependent increased levels of passive acetate uptake. Such more efficient substrate uptake could be related to a formation of calcium acetate complexes. Findings lead to bioprocess methods to stimulate a short-term selective growth of PHA-storing microorganisms and this enables improvements to the techno-economic feasibility for municipal waste activated sludge to become a generic resource for industrial scale PHA production.

The impact of acute lymphoblastic leukemia treatment on central nervous system results in Bogota, Colombia.

To improve the outcome of children with acute lymphoblastic leukemia (ALL) treated at the National Cancer Institute, Bogota, Colombia, a protocol based on the BFM-90 (Berlin, Frankfurt, Munster study) and the LSA2L2 regimens was implemented in the year 1993. The patients were classified as being standard risk (SR) or high risk (HR) according to clinical criteria, to which cytogenetic information and day-8 prednisone response were also added. A 123-patient cohort entered the study, 18 of them being considered SR and 105 HR. There was a 94% 10 years' event-free-survival rate for the SR group and 36% for the HR group. Decreased induction death rate (7% vs. 14%), increased complete remission (CR) rate (81% vs. 75%), and continuous CR (45% vs. 33%) were found in comparison with the previous study. A significant improvement was achieved in relapse rate, 44% to 28% (P=0.029), mainly due to reduced central nervous system relapse rate from 16% to 6% (P=0.037), whereas the number of patients receiving cranial radiation was reduced to 55%. A major problem concerned the increased CR mortality rate, 5% to 14% (P=0.06). Improved supportive care therapy and socioeconomic conditions will hopefully reduce the CR mortality rate in the future.

Chilblain lupus with nail involvement: a case report

Chilblain lupus is a rarely manifested variant of chronic lupus. Its appearance can be sporadic or hereditary associated with an autosomal dominant mutation. The diagnosis is clinical and histopathological. The case is presented of a patient with systemic lupus erythematosus presenting with chilblain lupus and nail involvement, despite the use of antimalarials and immunomodulators.

La perniosis lúpica es una variante del lupus crónico que se manifiesta con poca frecuencia, su aparición puede ser esporádica o hereditaria, asociada con una mutación autosómica dominante, en tanto que su diagnóstico es clínico e histopatológico. Se reporta el caso de una mujer con lupus eritematoso sistémico con manifestación de perniosis lúpica y compromiso ungueal, a pesar del uso de antimaláricos e inmunomoduladores.

The chronic obstructive pulmonary disease assessment test improves the predictive value of previous exacerbations for poor outcomes in COPD.

INTRODUCTION: Chronic obstructive pulmonary disease (COPD) exacerbations have a negative impact on the quality of life of patients and the evolution of the disease. We have investigated the prognostic value of several health-related quality of life questionnaires to predict the appearance of a composite event (new ambulatory or emergency exacerbation, hospitalization, or death) over a 1-year follow-up. METHODS: This was a multicenter, prospective, observational study. Patients completed four questionnaires after recovering from an exacerbation (COPD Assessment Test [CAT], a Clinical COPD Questionnaire [CCQ], COPD Severity Score [COPDSS], and Airways Questionnaire [AQ20]). Patients were followed-up until the appearance of the composite event or for 1 year, whichever came first. RESULTS: A total of 497 patients were included in the study. The majority of them were men (89.7%), with a mean age of 68.7 (SD 9.2) years, and a forced expiratory volume in 1 second of 47.1% (SD 17.5%). A total of 303 (61%) patients experienced a composite event. Patients with an event had worse mean scores of all questionnaires at baseline compared to patients without event: CAT=12.5 vs 11.3 (P=0.028); CCQ=2.2 vs 1.9 (P=0.013); COPDSS=12.3 vs 10.9 (P=0.001); AQ20=8.3 vs 7.5 (P=0.048). In the multivariate analysis, only previous history of exacerbations and CAT score ≥13.5 were significant risk factors for the composite event. A CAT score ≥13.5 increased the predictive value of previous exacerbations with an area under the receiver operating characteristic curve of 0.864 (95% CI: 0.829-0.899; P=0.001). CONCLUSION: The predictive value of previous exacerbations significantly increased only in one of the four trialled questionnaires, namely in the CAT questionnaire. However, previous history of exacerbations was the strongest predictor of the composite event.

Tratamiento multidisciplinario en una paciente con periodontitis agresiva generalizada y diabetes mellitus tipo 1 / Multi-disciplinary treatment of female patient afflicted with generalized aggressive periodontitis and type 1 diabetes mellitus

El impacto de las enfermedades sistémicas sobre la salud bucal se encuentra bien documentado. Algunos desórdenes sistémicos pueden modificar la respuesta inmune del huésped a los patógenos periodontales, exacerbando la severidad de la enfermedad periodontal. Dentro de las enfermedades sistémicas, la diabetes mellitus no controlada se encuentra asociada con la severidad de la enfermedad periodontal. La periodontitis agresiva se presenta en adultos jóvenes, causando una rápida destrucción del aparato de inserción periodontal. La pérdida severa del soporte periodontal presente en estos casos dificulta el pronóstico de los dientes y, por lo tanto, para el clínico se complica la planificación del tratamiento, y existe la posibilidad de extraer los dientes comprometidos. Para realizar un tratamiento integral es necesaria la participación de otras áreas odontológicas. El objetivo de este trabajo es presentar el tratamiento multidisciplinario en una paciente femenina de 17 años de edad con diabetes mellitus tipo 1 y periodontitis agresiva generalizada, y los resultados de su tratamiento a un año.

The impact of systemic diseases on oral health has been well documented. Certain systemic disorders can modify the host's immune response to periodontal pathogens, thus exacerbating the severity of the periodontal disease. Among systemic diseases, uncontrolled diabetes mellitus is associated to periodontal disease. Aggressive periodontitis can appear in young adults and elicit rapid destruction of the periodontal insertion apparatus. Severe loss of periodontal support present in these cases hinders prognosis of affected teeth, and thus, the clinician faces complications when designing treatment plans and deciding upon extraction or non extraction of compromised teeth. Accomplishment of comprehensive treatment requires participation of other fields of dentistry. The aim of the present study was to present the multi-disciplinary treatment of a 17-year-old female patient afflicted with type 1 diabetes mellitus and generalized aggressive periodontitis, and present results obtained one year after completion of treatment.

Tratamiento interdisciplinario de paciente con sonrisa gingival: Reporte de un caso / Interdisciplinary treatment of patient with gummy smile: Case report

Introducción: Un tratamiento integral bucal involucra la evaluación de las características faciales, actividad muscular, y la relación entre la dentición visible y los tejidos blandos para mejorar función y estética del paciente. Algunas condiciones fisiológicas y/o patológicas resultan en la exposición excesiva de la encía ocasionando un aspecto negativo, limitando la expresión facial. Una de las principales causas de una sonrisa gingival es la erupción pasiva alterada o retardada. El alargamiento de corona es una técnica quirúrgica muy valiosa para procedimientos restaurativos, diseñada para aumentar el tamaño de la corona clínica. Como parte de un manejo interdisciplinario, los implantes dentales son una opción de tratamiento cuando existen dientes ausentes. Caso clínico: Paciente femenina de 56 años acude al Departamento de Periodoncia e Implantología porque no le gusta la apariencia de sus dientes ni su sonrisa. A la exploración clínica se observa biotipo grueso, márgenes gingivales irregulares, coronas clínicas irregulares y cortas, ausencia dentales y dimensión vertical disminuida. Radiográficamente el margen óseo se encuentra a nivel de unión cemento esmalte. El diagnóstico es erupción pasiva alterada. Con el Departamento de Prótesis Bucal y con base al encerado diagnóstico se deciden hacer alargamientos de corona con cirugía ósea en dientes anteriores superiores e inferiores, colocación de implantes en zona de dientes 36 y 46 y rehabilitación protésica con el sistema Captek. Resultados: Los resultados fueron favorables, se logró obtener armonía dentofacial y dentoperiodontal, así como función, estética y satisfacción en la paciente.

Introduction: A comprehensive, inter-disciplinary treatment of the mouth involves the following aspects: assessment of facial characteristics and muscle activity as well as existing relationship between visible dentition and soft tissues to improve patients' aesthetics and function. There are some physiological and/or pathological conditions which result in excessive exposition of the gums. This causes a negative appearance and limits facial expression. Altered or delayed eruption is one of the main causes of gummy smile. In restorative procedures, crown elongation is considered a very valuable surgical technique. This technique has the aim of lengthening the size of the clinical crown. As part of an inter-disciplinary treatment, dental implants represent an option in cases where there are missing teeth. Clinical case: 56 year old female patient attended the Implantology and Periodontics Department complaining of not liking the appearance of her teeth and smile. Clinical exploration disclosed thick biotype, irregular gingival margins, short and uneven clinical crowns, missing teeth as well as decreased vertical dimension. X ray assessment revealed bony margin at the level of enamel and cement junction. Emitted diagnosis was: Altered Passive Eruption. Based on diagnostic wax-up and after consultation with the Oral Prosthesis Department, treatment selected was crown elongation with bone surgery in upper and lower front teeth, implant placement in areas of teeth 36 and 46 as well as rehabilitation following the Captek system. Results: Results were favorable. Dentofacial and dental-periodontal harmony were achieved, as well as function, aesthetics and the patient's satisfaction.