Cobalamin cbiP mutant shows decreased tolerance to low temperature and copper stress in Listeria monocytogenes.

BACKGROUND: Listeria monocytogenes is a foodborne pathogen that causes listeriosis in humans. This pathogen activates multiple regulatory mechanisms in response to stress, and cobalamin biosynthesis might have a potential role in bacterial protection. Low temperature is a strategy used in the food industry to control bacteria proliferation; however, L. monocytogenes can grow in cold temperatures and overcome different stress conditions. In this study we selected L. monocytogenes List2-2, a strain with high tolerance to the combination of low temperature + copper, to understand whether the cobalamin biosynthesis pathway is part of the tolerance mechanism to this stress condition. For this, we characterized the transcription level of three cobalamin biosynthesis-related genes (cbiP, cbiB, and cysG) and the eutV gene, a transcriptional regulator encoding gene involved in ethanolamine metabolism, in L. monocytogenes strain List2-2 growing simultaneously under two environmental stressors: low temperature (8 °C) + copper (0.5 mM of CuSO4 × 5H2O). In addition, the gene cbiP, which encodes an essential cobyric acid synthase required in the cobalamin pathway, was deleted by homologous recombination to evaluate the impact of this gene in L. monocytogenes tolerance to a low temperature (8 °C) + different copper concentrations. RESULTS: By analyzing the KEGG pathway database, twenty-two genes were involved in the cobalamin biosynthesis pathway in L. monocytogenes List2-2. The expression of genes cbiP, cbiB, and cysG, and eutV increased 6 h after the exposure to low temperature + copper. The cobalamin cbiP mutant strain List2-2ΔcbiP showed less tolerance to low temperature + copper (3 mM) than the wild-type L. monocytogenes List2-2. The addition of cyanocobalamin (5 nM) to the medium reverted the phenotype observed in List2-2ΔcbiP. CONCLUSION: These results indicate that cobalamin biosynthesis is necessary for L. monocytogenes growth under stress and that the cbiP gene may play a role in the survival and growth of L. monocytogenes List2-2 at low temperature + copper.

Consequences of multiple mating-system shifts for population and range-wide genetic structure in a coastal dune plant.

Evolutionary transitions from outcrossing to selfing can strongly affect the genetic diversity and structure of species at multiple spatial scales. We investigated the genetic consequences of mating-system shifts in the North American, Pacific coast dune endemic plant Camissoniopsis cheiranthifolia (Onagraceae) by assaying variation at 13 nuclear (n) and six chloroplast (cp) microsatellite (SSR) loci for 38 populations across the species range. As predicted from the expected reduction in effective population size (Ne ) caused by selfing, small-flowered, predominantly selfing (SF) populations had much lower nSSR diversity (but not cpSSR) than large-flowered, predominantly outcrossing (LF) populations. The reduction in nSSR diversity was greater than expected from the effects of selfing on Ne alone, but could not be accounted for by indirect effects of selfing on population density. Although selfing should reduce gene flow, SF populations were not more genetically differentiated than LF populations. We detected five clusters of nSSR genotypes and three groups of cpSSR haplotypes across the species range consisting of parapatric groups of populations that usually (but not always) differed in mating system, suggesting that selfing may often initiate ecogeographic isolation. However, lineage-wide genetic variation was not lower for selfing clusters, failing to support the hypothesis that selection for reproductive assurance spurred the evolution of selfing in this species. Within three populations where LF and SF plants coexist, we detected genetic differentiation among diverged floral phenotypes suggesting that reproductive isolation (probably postzygotic) may help maintain the striking mating-system differentiation observed across the range of this species.

Field Performance of Cryopreserved Seed-derived Maize Plants.

BACKGROUND: Cryo-preservation of plant materials in liquid nitrogen (LN) has been described as a suitable technology to conserve genetic resources of several species. However, the potential effects of LN in the subsequent plant growth in the field should be studied before large-scale implementation of cryopreserved germplasm banks. OBJECTIVE: To describe the field performance of cryopreserved seed-derived maize adult plants. MATERIALS AND METHODS: Germination percentage and numbers of leaves and ears per plant, internodes in stems, middle - aged leaf length, plant height, ear traits and weight of 100 seeds were recorded. RESULTS: Statistically significant differences between adult plants derived from cryopreserved seeds and the control treatment were not observed (t-test, p=0.05). CONCLUSION: The results presented confirm at the phenotype level the effectiveness of maize seed cryostorage to preserve and regenerate true-to-type plants.

Pro-inflammatory cytokines involvement in the hesperidin antihyperalgesic effects at peripheral and central levels in a neuropathic pain model.

Emerging evidence proposes a link between immune changes and pain, which is consistent with the inflammation theory and the increased incidence of neurodegenerative diseases. Flavonoids have long been used because of their anti-inflammatory potential activity and they are considered a promising alternative to alleviate neuropathic pain. The aim of this study was to investigate the antihyperalgesic effect of hesperidin and the presence of pro-inflammatory cytokines evaluated at peripheral and central levels in the chronic constriction injury as model of neuropathic pain in rats. Mechanical and thermal hyperalgesia were assessed in the aesthesiometer and plantar tests, respectively, as related to the presence of cytokines concentrations (TNF-α, IL-1ß and IL-6) in sciatic nerve and segments of the spinal cord after 15 days chronic constriction injury model in rats receiving vehicle or hesperidin. Antihyperalgesic response of hesperidin (100 mg/kg) was associated to the presence of cytokines mainly at several sections of the spinal cord suggesting not only peripheral but also its involvement in central sensitization in the experimental neuropathic pain.

Arthrodilation: a treatment algorithm as a conservative management option for adhesive capsulitis.

INTRODUCTION: the management of adhesive capsulitis (AC) remains a topic of debate among orthopedic surgeons, with a wide variation in the literature. Conservative treatment relies as the first-line option as clinical studies report positive outcomes. However, there is variability in the effectiveness of different treatment modalities. MATERIAL AND METHODS: this study aimed to analyzed functional and clinical outcomes of patients with AC who underwent the arthrodilation protocol, including three ultrasound-guided injections administered on a weekly basis: two corticosteroid injections and one injection of hyaluronic acid combined with corticosteroids. Additionally, patients received a specific rehabilitation therapy. Visual analogue scale (VAS), the university of California-Los Angeles shoulder score (UCLA) and Constant-Murley score were assessed before treatment and after 3-month follow-up period. RESULTS: 23 patients were included, receiving the same treatment protocol with a mean onset of symptoms of 4.9 ± 1.7 months. Among these patients, there was a clear predominance of females (65.2%). Age distribution ranged from 39 to 74 years (mean = 56) indicating that individuals in their mid-50s were more susceptible to developing this condition. Furthermore, a slight majority (52.2%) exhibited AC in their right shoulder. VAS significantly decreased (-6.09 ± 1.9 [p 0.05]). Similarly, UCLA score (10.9 ± 2.9 to 31.7 ± 2.2) and Constant-Murley score (22.3 ± 6.1 to 62.0 ± 6.2) improved significantly. Pre-to-post treatment evaluation showed improvement in both UCLA (mean = 20.8 ± 2.9 [p 0.05]) and Constant-Murley (mean = 39.7 ± 9 [p 0.05]). CONCLUSION: arthrodilation protocol demonstrated promising results, with patients achieving good to excellent outcomes and safely resuming their regular daily activities within a short-term follow-up period. These findings provide support for arthrodilation as a viable conservative management option and contribute valuable insights to the ongoing research aimed at identifying optimal treatment approaches for adhesive capsulitis.

INTRODUCCIÓN: el tratamiento de la capsulitis adhesiva (CA) sigue siendo un tema de debate entre los cirujanos ortopedistas, con una variación amplia en la literatura. El tratamiento conservador se considera la opción de primera línea, ya que los estudios clínicos muestran resultados positivos. Sin embargo, existe variabilidad en la efectividad de las diferentes modalidades de tratamiento. MATERIAL Y MÉTODOS: este estudio tuvo como objetivo analizar los resultados funcionales y clínicos de pacientes con CA que se sometieron al protocolo de artrodilatación, incluyendo tres inyecciones guiadas por ultrasonido administradas semanalmente: dos inyecciones de corticosteroides y una inyección de ácido hialurónico combinado con corticosteroides. Además, los pacientes recibieron una terapia de rehabilitación específica. La escala analógica visual (EVA), la puntuación del hombro de la Universidad de California-Los Ángeles (UCLA) y la puntuación de Constant-Murley se evaluaron antes del tratamiento y después de un período de seguimiento de tres meses. RESULTADOS: se incluyeron 23 pacientes que recibieron el mismo protocolo de tratamiento con un inicio medio de síntomas de 4.9 ± 1.7 meses. Entre estos pacientes hubo un claro predominio del sexo femenino (65.2%). La distribución por edades osciló entre 39 y 74 años (media = 56), lo que indica que las personas de alrededor de 55 años eran más susceptibles a desarrollar esta afección. Además, una ligera mayoría (52.2%) presentaba CA en el hombro derecho. La EVA disminuyó significativamente (-6.09 ± 1.9 [p 0.05]). De manera similar, la puntuación de UCLA (10.9 ± 2.9 a 31.7 ± 2.2) y la puntuación de Constant-Murley (22.3 ± 6.1 a 62.0 ± 6.2) mejoraron significativamente. La evaluación previa y posterior al tratamiento mostró una mejoría tanto en UCLA (media = 20.8 ± 2.9 [p 0.05]) como en Constant-Murley (media = 39.7 ± 9 [p 0.05]). CONCLUSIÓN: el protocolo de artrodilatación demostró resultados prometedores, los pacientes lograron resultados de buenos a excelentes y reanudaron de manera segura sus actividades diarias regulares dentro de un período de seguimiento a corto plazo. Estos hallazgos respaldan la artrodilatación como una opción de tratamiento conservador viable y aportan conocimientos valiosos a la continua investigación destinada a identificar tratamientos óptimos para la capsulitis adhesiva.

Clinical differences in a multidisciplinary pediatric pain unit between primary and secondary chronic pain.

INTRODUCTION: Chronic pain affects an important part of the pediatric population in developed countries. secondary chronic pain (SCP) can have a well-defined medical cause, but primary chronic pain (PCP) can have an unknown etiology. In Spain, there is as yet no information on the clinical differences between patients treated in multidisciplinary units. METHODS: Retrospective analysis of the clinical records of patients seen in 2018 at the Children's Chronic Pain Unit in University La Paz Hospital. RESULTS: A total of 92 patients were included, (age between 3 and 19 years), with a mean age of 12.4 (SD = 4.1) years, mostly female (55%), with a mean duration of pain of 11.3 (SD = 10.4) months. A comparison of patients with PCP (n = 31) and SCP (n = 61) showed that both groups, on average, presented intense pain (X = 5.9; SD = 2.2; range = 0-10), with similar duration and functional repercussions, although PCP was less likely to be associated with neuropathic descriptors than SCP (p = 0.040), and was more extensive (p < 0.001). Both groups received similar treatment, based on rehabilitation, psychotherapy, invasive techniques and analgesic medication, although patients in the PCP group received less analgesic medication (gabapentinoids and opioids) than the SCP (p = 0.011). CONCLUSION: Patients treated in a multidisciplinary Child Pain Unit for PCP or SCP present a very similar clinical profile, though with differences in the number and type of analgesic drugs used. This shows the importance of etiologic diagnosis for adequate pharmacological treatment.

Genetic characterization of Latin-American Creole cattle using microsatellite markers.

Genetic diversity in and relationships among 26 Creole cattle breeds from 10 American countries were assessed using 19 microsatellites. Heterozygosities, F-statistics estimates, genetic distances, multivariate analyses and assignment tests were performed. The levels of within-breed diversity detected in Creole cattle were considerable and higher than those previously reported for European breeds, but similar to those found in other Latin American breeds. Differences among breeds accounted for 8.4% of the total genetic variability. Most breeds clustered separately when the number of pre-defined populations was 21 (the most probable K value), with the exception of some closely related breeds that shared the same cluster and others that were admixed. Despite the high genetic diversity detected, significant inbreeding was also observed within some breeds, and heterozygote excess was detected in others. These results indicate that Creoles represent important reservoirs of cattle genetic diversity and that appropriate conservation measures should be implemented for these native breeds in order to minimize inbreeding and uncontrolled crossbreeding.

Early manifestations of autism spectrum disorders. Experience of 393 cases in a paediatric neurology.

INTRODUCTION: Autism spectrum disorders are group of conditions characterised by qualitative impairments in social communication, interaction, and imagination, and by a restricted range of interests and typical repetitive behaviours. Frequently, there is a delay in the age of detection, and therefore in starting multidisciplinary evaluations and interventions, which may result in a poorer prognosis and reduced quality of life for both children and parents. The aim of our study was to describe clinical and epidemiological data including the age of detection and main initial complaints present in children with autism disorders referred to a paediatric neurology centre. PATIENTS AND METHODS: A total of 393 medical records of consecutive cases diagnosed with an autism spectrum disorder were reviewed. RESULTS: Autism was diagnosed in 82.1% of the cases, unspecified pervasive disorder in 9.9%, Asperger syndrome in 4.8%, and Rett syndrome in 3%. Sixty percent of autistic children presented with a language disorder as their main complaint. The average age of detection was 4 years. CONCLUSIONS: Compared with other countries, age of detection is delayed. Primary care-based screening and surveillance are required in order to improve prognosis and quality of life of children with an autism spectrum disorder.

Hypertriglyceridemic waist: an alternative to the metabolic syndrome? Results of the IMAP Study (multidisciplinary intervention in primary care).

AIM: To study the prevalence of hypertriglyceridemic waist (HTGW) in an urban adult Spanish population and its association with type 2 diabetes mellitus (T2DM) and cardiovascular disease (CVD). METHODS: We undertook a cross-sectional analysis in a random sample of 2270 individuals (18-80 years of age). All participants provided a clinical history and underwent a physical examination. Blood and urine analyses were conducted. HTGW was diagnosed using anthropometric criteria for the European population (waist circumference: for men, ≥ 94 cm; for women, ≥ 80 cm) and fasting plasma triglycerides (TGs) ≥ 1.71 mmol l(-1) (≥ 150 mg per 100 ml). RESULTS: The prevalence of HTGW was 14.5% (men: 18.2%, women: 10.8%) and was significantly greater in men <59 years (P<0.001). HTGW was associated with older individuals, a low educational level and, in men, with a sedentary lifestyle (P<0.001). Subjects with HTGW had higher levels of total cholesterol, low-density lipoprotein-cholesterol (LDL-c) and uric acid, lower levels of high-density lipoprotein-cholesterol, a higher blood pressure, a greater degree of obesity and a higher prevalence of T2DM (20.00 vs 6.4%, P<0.001) (odds ratio (OR) 3.61; 95% confidence interval (95% CI), 2.60-5.01) and CVD (8.5 vs 3.4%, P<0.001) (OR 2.63; 95% CI, 1.66-4.16). The association of HTGW with T2DM and CVD disappeared after adjusting for age. The degree of concordance between HTGW and the metabolic syndrome (MS) was moderate, with both the Adult Treatment Panel III Report (ATP-III) and the International Diabetes Federation criteria (κ=0.51 and κ=0.58, respectively). Subjects with isolated HTGW as compared with those with isolated MS (ATP-III) were younger, had greater levels of total cholesterol, LDL-c and TGs and a lower prevalence of obesity, high blood pressure and dysglycemia. CONCLUSION: HTGW is a phenotype of cardiometabolic risk prevalent in the adult population in our environment. HTGW may be an alternative to MS to detect the population at risk for T2DM and CVD, especially in young individuals who do not fulfill the criteria for MS.

Pinus halepensis invasion in mountain pampean grassland: effects of feral horses grazing on seedling establishment.

We evaluated the mechanisms that determine the increasing abundance of Pinus halepensis in mountain pampean grasslands in Argentina that is associated with the continuous presence of feral horses. We hypothesized that direct and indirect effects of feral horse grazing on grassland may affect the establishment of P. halepensis. We conducted a field experiment to evaluate this hypothesis, studying the response of seedling emergence, survival and growth to herbaceous vegetation defoliation and soil disturbance in sites with contrasting grazing histories. We also evaluated the composition and structure of plant communities of each experimental site. Direct effects such as heavily defoliated perennial grasses and indirect effects such as reduced specific and functional richness, diversity and evenness enhanced the emergence, survival and early growth of the seedlings. High bare ground percentage and low grass biomass also positively affected seedling establishment. The experimental evidence demonstrates that the combination of invasional meltdown hypothesis and weakened biotic resistance are the mechanisms involved in P. halepensis invasion.

Comparing the activPAL software's Primary Time in Bed Algorithm against Self-Report and van der Berg's Algorithm.

The purpose of this study was to compare activPAL algorithm-estimated values for time in bed (TIB), wake time (WT) and bedtime (BT) against self-report and an algorithm developed by van der Berg and colleagues. Secondary analyses of baseline data from the Community Activity for Prevention Study (CAPS) were used in which adults ≥ 18 years wore the activPAL for seven days. Mixed-effects models compared differences between TIB, WT, and BT for all three methods. Bland-Altman plots examined agreement and the two-one-sided test examined equivalence. activPAL was not equivalent to self-report or van der Berg in estimating TIB, but was equivalent to self-report for estimating BT, and was equivalent to van der Berg for estimating WT. The activPAL algorithm requires adjustments before researchers can use it to estimate TIB. However, researchers can use activPAL's option to manually enter self-reported BT and WT to estimate TIB and better understand 24-hour movement patterns.

[Fixation of acetabulum posterior column fracture, arthroscopy assisted]. / Fijación de fractura de columna posterior de acetábulo, asistida por artroscopía.

Arthroscopic surgery of the hip has evolved significantly over the last years, offering an opportunity of treatment in several intra-articular pathologies. Recent methods and instrumental advances have allowed more frequent use of this procedure, with various indications such as: hip pain, acetabular labrum lesions, extraction of foreign bodies, debridement in septic arthritis, pincer impingement, and some extra-articular injuries. Another intra-articular hip pathology that benefits from arthroscopic assistance could be found in the management of simple acetabular fractures. Therefore, this report presents the case of a patient with a posterior column fracture treated with an arthroscopic approach. Arthroscopic techniques for the fixation of intra-articular fractures offer the advantage of a direct visualization of the articular surface, which results in an anatomical reduction. In addition, arthroscopic techniques also present a benefit for intra-articular injuries such as the posterior labral lesion in this case. In simple fractures of the posterior acetabular column with minimum displacement, where percutaneous screw fixation is useful as a definitive fixation method, hip arthroscopy is useful for the reduction process and verifies the extra-articular screw trajectory.

La cirugía artroscópica de la cadera ha evolucionado considerablemente en los últimos años, ofreciendo oportunidad de tratamiento a muchas patologías intraarticulares no reconocidas previamente. Con avances recientes en métodos e instrumentos quirúrgicos, se realiza cada vez más frecuentemente, con indicaciones que incluyen: diagnóstico de pacientes con dolor de cadera, lesiones del labrum acetabular, extracción de cuerpos extraños, desbridamiento en artritis séptica, pinzamiento femoroacetabular y algunos daños extraarticulares. Otra patología intraarticular de la cadera que se beneficia de la asistencia por artroscopía se podría encontrar en el manejo de fracturas acetabulares simples. Reportamos el caso de un paciente con fractura de columna posterior acetabular, tratado vía artroscópica, y los resultados obtenidos. Las técnicas artroscópicas para la fijación de fracturas intraarticulares ofrecen la ventaja de una visualización y reducción superiores de la superficie articular, ya que se hacen mediante observación del foco de fractura, permitiendo una reducción anatómica. Además, las técnicas de reducción artroscópica ofrecen otro beneficio para las lesiones intraarticulares, como en este caso la lesión de labrum posterior. En los trazos simples con poco desplazamiento de las fracturas de columna posterior acetabular, donde la fijación percutánea con tornillos es útil como método de fijación definitiva, la artroscopía de cadera es de utilidad con el fin de asistir durante el proceso de reducción y verificar que el trayecto del tornillo no se encuentre intraarticular.

CAPS on the move: Crafting an approach to recruitment for a randomized controlled trial of community gardening.

OBJECTIVE: To describe and evaluate recruitment approaches for a randomized controlled trial (RCT) of community gardening in Denver, Colorado. (ClinicalTrials.gov: NCT03089177). METHODS: We used community and staff feedback to adapt our recruitment approach from year 1 to year 2 of a multi-year RCT to address health behaviors related to cancer prevention. In year 2, we added a full-time recruitment coordinator, designed and implemented a tracking spreadsheet, and engaged advisory committee members, local garden leaders, and health partners in planning and outreach. Screening and consent rates, staff time and costs for years 1 and 2 are compared. RESULTS: In year 1, recruitment methods yielded 136 initial contacts, 106 screenings and 64 consented participants. In year 2, enhanced staffing and outreach yielded 257 initial contacts, 193 screenings, and 123 consented participants. Personal referrals, health fairs, NextDoor, and fliers yielded the highest percentage of consented participants. School and community meetings yielded the lowest yield for potential participants. Spanish-speaking participants were mostly recruited by direct methods. Compared to year 1 recruitment, which required 707 h of staff time and cost $14,446, year 2 recruitment required 1224 h of staff time and cost $22,992. Average cost for retained participants was $226 (year 1) and $186 (year 2). DISCUSSION: Those planning pragmatic clinical trials with recruitment in multi-ethnic communities can use the results from this study to understand the efficacy of techniques, and to budget costs for recruitment. While our culturally-tailored recruitment methods cost more, they provided more effective and efficient ways to reach recruitment goals.

[Membrane induction and transposition of non-vascularized ipsilateral fibula in posttraumatic bone neoformation. Case report]. / Inducción de membrana y trasposición de peroné ipsolateral no vascularizado en neoformación ósea postraumática. Reporte de caso.

OBJECTIVE: To demonstrate the outcome of tibial bone neo formation, using induced-membrane technique and non-vascularized ipsolateral fibular graft transposition. CASE REPORT: A 25 years old male with a 2 years ago firearm injury in left leg, presenting an initial diagnosis of open fracture Gustilo IIIB AO 42C3 IO4NV1MT2 with a 7cm tibial diaphyseal bone defect. During his hospital evolution multiple interventions were made including surgical debridement and skin grafts placement, with unfavorable results. Therefore, we decided to use the induced-membrane technique and non-vascularized ipsolateral fibular graft transposition, resulting in a cane dependent ambulation, in 4 months evolution after last intervention. CONCLUSION: Induced-membrane technique and non-vascularized ipsolateral fibular graft transposition could be a successful alternative for the management of patients with severe bone loss.

OBJETIVO: Demostrar los resultados en la neoformación ósea de tibia utilizando la técnica de inducción de membrana más la transposición de peroné ipsolateral no vascularizado. CASO CLÍNICO: Masculino de 25 años, antecedente de sufrir impacto por arma de fuego en pierna izquierda dos años antes, presentó un diagnóstico inicial de fractura expuesta Gustilo IIIB AO 42C3 IO4NV1MT2 con pérdida ósea de 7 cm de diáfisis tibial. Durante su evolución se realizaron múltiples aseos quirúrgicos y colocación de injertos cutáneos con resultados no favorables, por lo que se emplea la técnica de membrana inducida más colocación de peroné ipsolateral no vascularizado, teniendo una evolución a los cuatro meses posterior a su última intervención con deambulación dependiente de bastón. CONCLUSIÓN: La técnica de inducción de membrana con transposición de peroné ipsolateral no vascularizado podría ser una alternativa adecuada para el manejo de pérdidas óseas en los pacientes.

Rationale and design for the community activation for prevention study (CAPs): A randomized controlled trial of community gardening.

BACKGROUND: Engaging in health-promoting behaviors (e.g., healthy fruit- and vegetable-rich diet, physical activity) and living in supportive social and built environments are consistently and significantly associated with reductions in cancer, heart disease, diabetes, and other chronic diseases. Interventions to change diet and physical activity behaviors should aim to educate individuals, change the environments in which people live, work and recreate, improve access, availability, and affordability of healthy foods, and create safe places the facilitate active lifestyles. This trial will assess whether community gardening increases fruit and vegetable consumption and physical activity, improves social support and mental health, and reduces age-associated weight gain and sedentary time among a multi-ethnic, mixed-income population. METHODS/DESIGN: A randomized controlled trial of community gardening began in Denver, Colorado in January 2017. Over 3 years, we will recruit 312 consenting participants on Denver Urban Gardens' waitlists and randomize them to garden or remain on the waitlist. At baseline (pre-gardening), harvest time, and post-intervention, study participants will complete three 24-hour dietary recalls, a 7-day activity monitoring period using accelerometry, a health interview and physical anthropometry. DISCUSSION: This project addresses health-promoting behaviors among a multi-ethnic, mixed-income adult population in a large metropolitan area. If successful, this trial will provide evidence that community gardening supports and sustains healthy and active lifestyles, which can reduce risk of cancer and other chronic diseases. TRIAL REGISTRATION: ClinicalTrials.gov, ID: NCT03089177: Registered on 03/17/17.

Cobalamin cbiP mutant shows decreased tolerance to low temperature and copper stress in Listeria monocytogenes

BACKGROUND: Listeria monocytogenes is a foodborne pathogen that causes listeriosis in humans. This pathogen activates multiple regulatory mechanisms in response to stress, and cobalamin biosynthesis might have a potential role in bacterial protection. Low temperature is a strategy used in the food industry to control bacteria proliferation; however, L. monocytogenes can grow in cold temperatures and overcome different stress conditions. In this study we selected L. monocytogenes List2-2, a strain with high tolerance to the combination of low temperature +copper, to understand whether the cobalamin biosynthesis pathway is part of the tolerance mechanism to this stress condition. For this, we characterized the transcription level of three cobalamin biosynthesis related genes ( cbiP , cbiB, and cysG ) and the eutV gene, a transcriptional regulator encoding gene involved in ethanolamine metabolism, in L. monocytogenes strain List2-2 growing simultaneously under two environmental stressors: low temperature (8 °C) +copper (0.5 mM of CuSO4 ×5H2O). In addition, the gene cbiP , which encodes an essential cobyric acid synthase required in the cobalamin pathway, was deleted by homologous recombination to evaluate the impact of this gene in L. monocytogenes tolerance to a low temperature (8 °C) +different copper concentrations. RESULTS: By analyzing the KEGG pathway database, twenty-two genes were involved in the cobalamin biosynthesis pathway in L. monocytogenes List2-2. The expression of genes cbiP , cbiB, and cysG, and eutV increased 6 h after the exposure to low temperature +copper. The cobalamin cbiP mutant strain List2-2Δ cbiP showed less tolerance to low temperature +copper (3 mM) than the wild type L. monocytogenes List2-2. The addition of cyanocobalamin (5 nM) to the medium reverted the phenotype observed in List2-2Δ cbiP . CONCLUSION: These results indicate that cobalamin biosynthesis is necessary for L. monocytogenes growth under stress and that the cbiP gene may play a role in the survival and growth of L. monocytogenes List2-2 at low temperature +copper.

Association of differing dietary, metabolic, and clinical risk factors with microvascular complications of diabetes: a prevalence study of 503 Mexican type II diabetic subjects. II.

The overall prevalences of microvascular complications and their association with dietary, clinical, and metabolic characteristics have been studied in 503 Mexican type II diabetic subjects. Average daily dietary intakes were 1866 kcal, 46.5% as carbohydrate, 13.7 mmol cholesterol, 8.7 g fiber, and a polyunsaturated/saturated fat ratio of 0.98. Prevalence rates of microvascular and metabolic complications were as follows: background retinopathy 12.3%, exudative retinopathy 24.2%, proliferative retinopathy 8.1% (1% blind, 4% able to read large print only), 9.1% of patients had cataract, 15.9% nephropathy, and 40.8% peripheral neuropathy. In addition, 3.6% had experienced transient lower motor neuron facial paralysis and 0.2% oculomotor paralysis. Patients with retinopathy had a longer mean duration of diabetes, were less obese at the time of examination, and had higher initial and mean blood pressures and higher mean fasting blood glucose levels when compared with those without retinopathy. Similar differences were observed between groups with and without nephropathy except that mean blood glucose levels were similar in the two groups. The presence of peripheral neuropathy was associated with longer duration of diabetes, less obesity, higher mean blood pressure and mean blood glucose levels, and lower hemoglobin concentration. Patients treated with diet alone had significantly lower prevalences of all three microvascular complications but they also had significantly shorter duration of diabetes and lower mean blood glucose levels. However, multivariate analyses on the subgroup of 360 patients who had repeated fasting blood glucose measurements for at least 5 yr demonstrated associations between retinopathy and duration of diabetes, mean blood pressure and mean blood glucose, and percent calories from carbohydrate.(ABSTRACT TRUNCATED AT 250 WORDS)

Association of differing dietary, metabolic, and clinical risk factors with macrovascular complications of diabetes: a prevalence study of 503 Mexican type II diabetic subjects. I.

Macrovascular and microvascular complications of diabetes may be associated with different environmental factors. To investigate this further, a prevalence study of 503 Mexican type II diabetic subjects was carried out while their patterns of nutrition were constrained by government food subsidies. Average daily dietary intakes were 1866 kcal; 46.5% as carbohydrate, 13.7 mmol cholesterol, 8.7 g fiber, and a polyunsaturated/saturated fat ratio of 0.98. With respect to macrovascular disease, 49.3% of patients had evidence of peripheral vascular disease, and 21.6% myocardial ischemia, 6.0% angina, 10.8% EKG evidence of ischemia, 4.8% EKG evidence of myocardial infarction. Only 1.2% (six patients) had a clear history of completed stroke, and all were hypertensive. Six patients had also undergone amputations for diabetic gangrene. Tabulation of the means of clinical characteristics according to presence or absence of myocardial ischemia showed that higher cholesterol, calorie, and fat intake, higher mean blood pressure, higher serum cholesterol, and serum triglyceride levels were found in those with myocardial ischemia. Patients with peripheral vascular disease were more commonly smokers. Stepwise logistic regression revealed significant positive associations between myocardial ischemia and dietary cholesterol, serum cholesterol, and mean blood pressure. In contrast, the presence of peripheral vascular disease was significantly related only to smoking and retinopathy. There were no associations between macrovascular complications and duration of diabetes in the multivariate analysis, and they occurred with equal frequency in men and women. Prospective studies of atherosclerosis in maturity-onset diabetes should assess and seek to modify dietary cholesterol, serum cholesterol, and hypertension.

Trend and seasonality in hospitalizations for pulmonary embolism: a time-series analysis.

BACKGROUND: The existence of seasonal variability in patients with acute pulmonary embolism (PE) has been debated for years, with contradictory results. The aim of this study was to identify the trend and possible existence of a seasonal pattern in hospitalizations for PE in Spain. METHODS: We analyzed the hospital discharge database of the Spanish National Health System from 2001 to 2010. Patients aged > 14 years diagnosed with PE were selected and a time series was constructed considering mean daily admissions for PE by month. The trend and seasonality factor of the series were determined using time-series analysis, and time-series modeling was used for analysis. Exponential smoothing models and the autoregressive integrated moving average test were used to generate a predictive model. RESULTS: From 2001 to 2010, there were 162,032 diagnoses of PE (5.07 per 1000 hospitalizations). In 105,168 cases, PE was the reason for admission. The PE diagnosis rate ranged from 4.14 per 1000 in 2001 to 6.56 per 1000 in 2010; and hospital admissions due to PE ranged from 2.67 to 4.28 per 1000 hospital discharges. Time-series analysis showed a linear increase in the incidence and a significant seasonal pattern with 17% more admissions in February and 12% fewer in June-July with respect to the central tendency (difference from February to June, 29%). CONCLUSIONS: The incidence of hospitalizations for PE showed a linear increase and a seasonal pattern, with the highest number of admissions in winter and the lowest number in summer.

Novel benzisoxazole derivatives as potent and selective inhibitors of acetylcholinesterase.

A series of N-benzylpiperidine benzisoxazoles has been developed as potent and selective inhibitors of the enzyme acetylcholinesterase (AChE). The benzisoxazole heterocycle was found to be an appropriate bioisosteric replacement for the benzoyl functionality present in the N-benzylpiperidine class of inhibitors. The title compounds were synthesized by alkylating 3-methyl-1,2-benzisoxazoles with an iodo piperidine derivatives as the key step. Benzisoxazoles 1b-j,o displayed potent inhibition of AChE in vitro with IC50's = 0.8-14 nM. Particularly interesting were N-acetyl and morpholino derivatives 1g (IC50 = 3 nM) and 1j (IC50 = 0.8 nM), respectively, which displayed outstanding selectivity for acetyl-over butyrylcholinesterase, in excess of 3 orders of magnitude. N-Acetyl 1g also displayed a favorable profile in vivo. This analog showed a dose-dependent elevation of total acetylcholine in mouse forebrain after oral administration with an ED50 = 2.4 mg/kg. In addition, 1g was able to reverse amnesia in a mouse passive avoidance model at doses of 3.2 and 5.6 mg/kg with an average reversal of 89.7%. Molecular dynamics simulations were used to study the possible binding modes of N-benzylpiperidine benzisoxazoles to AChE from Torpedo californica. Key structural insights were obtained regarding the potency of this class of inhibitors. Specifically, Asp-72, Trp-84, Trp-279, Phe-288, and Phe-330 are implicated in the binding of these inhibitors. The N-benzylpiperidine benzisoxazoles may be suitable compounds for the palliative treatment of Alzheimer's Disease.