The Emerging Role of 3-Hydroxyanthranilic Acid on C. elegans Aging Immune Function.

3-hydroxyanthranilic acid (3HAA) is considered to be a fleeting metabolic intermediate along tryptophan catabolism through the kynurenine pathway. 3HAA and the rest of the kynurenine pathway have been linked to immune response in mammals yet whether it is detrimental or advantageous is a point of contention. Recently we have shown that accumulation of this metabolite, either through supplementation or prevention of its degradation, extends healthy lifespan in C. elegans and mice, while the mechanism remained unknown. Utilizing C. elegans as a model we investigate how 3HAA and haao-1 inhibition impact the host and the potential pathogens. What we find is that 3HAA improves host immune function with aging and serves as an antimicrobial against gram-negative bacteria. Regulation of 3HAA's antimicrobial activity is accomplished via tissue separation. 3HAA is synthesized in the C. elegans hypodermal tissue, localized to the site of pathogen interaction within the gut granules, and degraded in the neuronal cells. This tissue separation creates a new possible function for 3HAA that may give insight to a larger evolutionarily conserved function within the immune response.

On the benefits of the tryptophan metabolite 3-hydroxyanthranilic acid in Caenorhabditis elegans and mouse aging.

Tryptophan metabolism through the kynurenine pathway influences molecular processes critical to healthy aging including immune signaling, redox homeostasis, and energy production. Aberrant kynurenine metabolism occurs during normal aging and is implicated in many age-associated pathologies including chronic inflammation, atherosclerosis, neurodegeneration, and cancer. We and others previously identified three kynurenine pathway genes-tdo-2, kynu-1, and acsd-1-for which decreasing expression extends lifespan in invertebrates. Here we report that knockdown of haao-1, a fourth gene encoding the enzyme 3-hydroxyanthranilic acid (3HAA) dioxygenase (HAAO), extends lifespan by ~30% and delays age-associated health decline in Caenorhabditis elegans. Lifespan extension is mediated by increased physiological levels of the HAAO substrate 3HAA. 3HAA increases oxidative stress resistance and activates the Nrf2/SKN-1 oxidative stress response. In pilot studies, female Haao knockout mice or aging wild type male mice fed 3HAA supplemented diet were also long-lived. HAAO and 3HAA represent potential therapeutic targets for aging and age-associated disease.

Long-Term Culture of Individual Caenorhabditis elegans on Solid Media for Longitudinal Fluorescence Monitoring and Aversive Interventions.

Caenorhabditis elegans are widely used to study aging biology. The standard practice in C. elegans aging studies is to culture groups of worms on solid nematode growth media (NGM), allowing the efficient collection of population-level data for survival and other physiological phenotypes, and periodic sampling of subpopulations for fluorescent biomarker quantification. Limitations to this approach are the inability to (1) follow individual worms over time to develop age trajectories for phenotypes of interest and (2) monitor fluorescent biomarkers directly in the context of the culture environment. Alternative culture approaches use liquid culture or microfluidics to monitor individual animals over time, in some cases including fluorescence quantification, with the tradeoff that the culture environment is contextually distinct from solid NGM. The WorMotel is a previously described microfabricated multi-well device for culturing isolated worms on solid NGM. Each worm is maintained in a well containing solid NGM surrounded by a moat filled with copper sulfate, a contact repellent for C. elegans, allowing longitudinal monitoring of individual animals. We find copper sulfate insufficient to prevent worms from fleeing when subjected to aversive interventions common in aging research, including dietary restriction, pathogenic bacteria, and chemical agents that induce cellular stress. The multi-well devices are also molded from polydimethylsiloxane, which produces high background artifacts in fluorescence imaging. This protocol describes a new approach for culturing isolated roundworms on solid NGM using commercially available polystyrene microtrays, originally designed for human leukocyte antigen (HLA) typing, allowing the measurement of survival, physiological phenotypes, and fluorescence across the lifespan. A palmitic acid barrier prevents worms from fleeing, even in the presence of aversive conditions. Each plate can culture up to 96 animals and easily adapts to a variety of conditions, including dietary restriction, RNAi, and chemical additives, and is compatible with automated systems for collecting lifespan and activity data.

Outbreak-Related Disease Burden Associated with Consumption of Unpasteurized Cow's Milk and Cheese, United States, 2009-2014.

The growing popularity of unpasteurized milk in the United States raises public health concerns. We estimated outbreak-related illnesses and hospitalizations caused by the consumption of cow's milk and cheese contaminated with Shiga toxin-producing Escherichia coli, Salmonella spp., Listeria monocytogenes, and Campylobacter spp. using a model relying on publicly available outbreak data. In the United States, outbreaks associated with dairy consumption cause, on average, 760 illnesses/year and 22 hospitalizations/year, mostly from Salmonella spp. and Campylobacter spp. Unpasteurized milk, consumed by only 3.2% of the population, and cheese, consumed by only 1.6% of the population, caused 96% of illnesses caused by contaminated dairy products. Unpasteurized dairy products thus cause 840 (95% CrI 611-1,158) times more illnesses and 45 (95% CrI 34-59) times more hospitalizations than pasteurized products. As consumption of unpasteurized dairy products grows, illnesses will increase steadily; a doubling in the consumption of unpasteurized milk or cheese could increase outbreak-related illnesses by 96%.

Green Tea Extract and Catechol-O-Methyltransferase Genotype Modify Fasting Serum Insulin and Plasma Adiponectin Concentrations in a Randomized Controlled Trial of Overweight and Obese Postmenopausal Women.

BACKGROUND: Green tea consumption has been associated with favorable changes in body weight and obesity-related hormones, although it is not known whether these changes result from green tea polyphenols or caffeine. OBJECTIVE: We examined the impact of decaffeinated green tea extract (GTE) containing 843 mg of (-)-epigallocatechin-3-gallate on anthropometric variables, obesity-associated hormones, and glucose homeostasis. METHODS: The Minnesota Green Tea Trial was a 12-mo randomized, double-blind, placebo-controlled clinical trial of 937 healthy postmenopausal women assigned to either decaffeinated GTE (1315 mg total catechins/d) or a placebo, stratified by catechol-O-methyltransferase (COMT) genotype. This study was conducted in a subset of 237 overweight and obese participants [body mass index (BMI) ≥25 kg/m(2)]. RESULTS: No changes in energy intake, body weight, BMI, or waist circumference (WC) were observed over 12 mo in women taking GTE (n = 117) or placebo (n = 120). No differences were seen in circulating leptin, ghrelin, adiponectin, or glucose concentrations at month 12. Participants randomly assigned to GTE with baseline insulin ≥10 µIU/mL (n = 23) had a decrease in fasting serum insulin from baseline to month 12 (-1.43 ± 0.59 µIU/mL), whereas those randomly assigned to placebo with baseline insulin ≥10 µIU/mL (n = 19) had an increase in insulin over 12 mo (0.55 ± 0.64 µIU/mL, P < 0.01). Participants with the homozygous high-activity (G/G) form of COMT had significantly lower adiponectin (5.97 ± 0.50 compared with 7.58 ± 0.53 µg/mL, P = 0.03) and greater insulin concentrations (7.63 ± 0.53 compared with 6.18 ± 0.36 µIU/mL, P = 0.02) at month 12 compared with those with the low-activity (A/A) genotype, regardless of treatment group. CONCLUSIONS: Decaffeinated GTE was not associated with reductions in body weight, BMI, or WC and did not alter energy intake or mean hormone concentrations in healthy postmenopausal women over 12 mo. GTE decreased fasting insulin concentrations in those with elevated baseline fasting concentrations. The high-activity form of the COMT enzyme may be associated with elevations in insulin and a reduction in adiponectin concentrations over time. This trial was registered at http://www.clinicaltrials.gov as NCT00917735.

Long-Term Supplementation of Green Tea Extract Does Not Modify Adiposity or Bone Mineral Density in a Randomized Trial of Overweight and Obese Postmenopausal Women.

BACKGROUND: Green tea extract (GTE) consumption has been linked to favorable changes in adiposity and bone mineral density (BMD), although it is unknown if these effects are due to green tea catechins or caffeine. The catechol-O-methyltransferase (COMT) genotype may also modify these associations. OBJECTIVE: We examined the impact of decaffeinated GTE on body composition (using dual-energy X-ray absorptiometry) and obesity-associated hormones. METHODS: The Minnesota Green Tea Trial was a 12-mo randomized, double-blind, placebo-controlled clinical trial in 937 postmenopausal women (aged 50-70 y) assigned to receive either GTE containing 843 mg (-)-epigallocatechin-3-gallate or placebo. This substudy was conducted in 121 overweight/obese participants [body mass index (BMI) (kg/m(2)) ≥25.0]. RESULTS: There were no differences in changes in BMI (-0.13 ± 0.11 compared with -0.05 ± 0.11; P = 0.61), total fat mass (-0.30 ± 0.16 compared with -0.12 ± 0.15 kg; P = 0.40), percentage of body fat (-0.15% ± 0.17% compared with -0.15% ± 0.16%; P = 0.99), or BMD (-0.006 ± 0.002 compared with -0.003 ± 0.002 g/cm(2); P = 0.49) over 12 mo between women taking GTE (n = 61) and those taking a placebo (n = 60). Interactions were observed between treatment and time for gynoid percentage of fat (%fat) and tissue %fat. Gynoid %fat increased from baseline to month 12 in the placebo group as baseline BMI increased and decreased over time as baseline BMI increased in the GTE group (P-interaction = 0.02). Tissue %fat increased from baseline to month 12 in the placebo group as baseline BMI increased. In the GTE group, tissue %fat decreased during the intervention as baseline BMI increased (P-interaction = 0.04). No changes were seen in circulating leptin, ghrelin, adiponectin, or insulin concentrations. COMT genotype did not modify the effect of GTE on any variable. CONCLUSIONS: Decaffeinated GTE was not associated with overall reductions in adiposity or improvements in BMD in overweight/obese postmenopausal women. However, GTE may be beneficial for reduction in tissue and gynoid %fat in individuals with higher BMI. This clinical trial was registered at www.clinicaltrials.gov as NCT00917735.

Trastornos del vínculo: detección y tratamiento / Attachment disorders: detection and treatment

El presente artículo describe una revisión panorámica y breve de la variedad de programas dirigidos a la promoción e intervención en apego y sensibilidad, desde un enfoque promocional, preventivo y de tratamiento. En primer lugar se revisan las conceptualizaciones dominantes que definen los trastornos del apego y a continuación se resumen los diferentes focos de trabajo de una muestra representativa de programas de intervención en apego. La mayoría de las intervenciones se focalizan en la interacción madre-hijo, en general están diseñadas para infantes y niños menores de tres años y se centran en mejorar las capacidades de sensibilidad parental así como los esquemas representativos de los padres en relación al hijo. Se concluye una creciente cantidad de propuestas con un enfoque de tipo universal e indicado, para díadas con problemas precoces de interacciones poco sensibles y condiciones psicosociales de cierto riesgo para apego inseguro. A su vez se describen intervenciones selectivas para poblaciones con presencia de maltrato y trastornos graves del apego. Se describen brevemente tres modalidades de intervención y sus rasgos, que representan diferentes focos y objetivos. Se discute finalmente sobre el tópico de la evidencia de efectividad y los factores asociados.

This article describes a panoramic review of the variety of programs aimed at the promotion and intervention in attachment and sensitivity from a promotional, preventive approach and treatment. First, the dominant conceptualizations of attachment disorders are reviewed, and then the different foci of work of a representative sample of attachment intervention programs are summarized. Most interventions are located in the mother-child interaction, are generally designed for infants and children under three years, and focus on improving the capabilities of parental sensitivity and representational schemes of parents regarding the child. We conclude that there are a growing number of proposals with a universal preventive approach for dyads with early attachment problems and insensitive interactions and psychosocial conditions that are risk factors for insecure attachment. At the same time we describe interventions for populations with severe abuse and attachment disorders. Three modes of intervention and their features are briefly described, representing different foci and objectives. We finally discuss the topic of evidence of effectiveness and associated factors.

Effects of tylosin administration on C-reactive protein concentration and carriage of Salmonella enterica in pigs.

OBJECTIVE: To evaluate the effects of tylosin on C-reactive protein concentration, carriage of Salmonella enterica, and antimicrobial resistance genes in commercial pigs. ANIMALS: 120 pigs on 2 commercial farms. PROCEDURES: A cohort of sixty 10-week-old pigs in 4 pens/farm (15 pigs/pen) was randomly selected. Equal numbers of pigs were given feed containing tylosin (40 µg/g of feed) for 0, 6, or 12 weeks. C-reactive protein concentrations were measured, microbial culture for S enterica in feces was performed, and antimicrobial resistance genes in feces were quantified. RESULTS: No significant associations were detected between C-reactive protein concentration or S enterica status and tylosin treatment. During the 12 weeks of tylosin administration, increased levels of 6 antimicrobial resistance genes did not occur. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of pigs with tylosin did not affect C-reactive protein concentration or reduce carriage or load of S enterica. There was no evidence that pigs receiving tylosin had increased carriage of the 6 antimicrobial resistance genes measured. IMPACT FOR HUMAN MEDICINE: S enterica is a public health concern. Use of the antimicrobial growth promoter tylosin did not pose a public health risk by means of increased carriage of S enterica.

Effect of delayed exposure of cattle to Mycobacterium avium subsp paratuberculosis on the development of subclinical and clinical Johne's disease.

OBJECTIVE: To evaluate the effect of delayed exposure of dairy cattle to Mycobacterium avium subsp paratuberculosis (MAP) on the incidence of those cows testing positive for MAP and developing clinical Johne's disease (CJD). ANIMALS: 79 cows not exposed to MAP as calves (unexposed cohort) and 260 cows exposed to MAP as calves (exposed cohort). PROCEDURES: Cows in the unexposed cohort were born into 5 MAP-uninfected herds and introduced at various ages into 5 MAP-infected herds where the exposed cohort cows were born and raised. Beginning when each cow was 24 months old, fecal and serum samples were collected annually from 2003 through 2006. Feces were cultured for MAP, and an ELISA was used to analyze serum samples for antibodies against MAP. Date and reason for culling were obtained from herd records. Incidence of positive culture and ELISA results and CJD was compared between unexposed and exposed cohort cows with Cox regression. RESULTS: Compared with exposed cohort cows, the hazard ratios for unexposed cohort cows having positive culture results, having positive ELISA results, and developing CJD were 0.12, 0.03, and 0.001, respectively, and those ratios increased by 2%, 6%, and 17%, respectively, for each month spent in an MAP-infected herd. CONCLUSIONS AND CLINICAL RELEVANCE: Delayed exposure of cows to MAP resulted in lower incidences of positive culture and ELISA results and CJD in those cows, compared with incidences of cows exposed to MAP since birth. The hazard of testing positive for MAP or developing CJD increased with time, regardless of cohort.

Is an individual calving pen better than a group calving pen for preventing transmission of Mycobacterium avium subsp paratuberculosis in calves? Results from a field trial.

The objective of this study was to quantify the efficacy of using individual calving pens (ICP) from which manure was removed between successive calving compared with group calving pens (GCP) for limiting transmission of Mycobacterium avium subsp paratuberculosis (MAP) in Holstein calves. Every other pregnant cow in three Minnesota MAP endemic herds was assigned to calve in either the ICP or the GCP within 48-72 h prior to expected calving. Heifer calves born in the ICP were assigned to the intervention group (n=238) while heifer calves born in the GCP were considered controls (n=211). Calves were separated from their dams as soon as was possible once the calf was found. The intervention within the ICP relative to the GCP was the removal of fecal material in the ICP immediately after each birth. Upon enrollment in 2005, calves were monitored into adulthood. Of the original animals enrolled, 318 were tested for MAP at least once in 2007, 2009, or 2010 using serum ELISA (ICP, n=165; GCP, n=141) and bacterial culture of feces (ICP, n=173; GCP, n=145) tests. Cox regression analysis was performed to evaluate the time until MAP test positivity. Cows born in the ICP had a hazard ratio of 0.37 (95% CI=0.34-0.4) for testing MAP serum ELISA positive, compared with cows born in GCP. Similarly, cows born in the ICP had a hazard ratio of 0.09 (95% CI=0.06-0.14) for testing MAP fecal culture positive, compared with cows born in GCP. The Cox proportional-hazard assumption was violated in both models such that differences observed in the instantaneous hazards of MAP positive outcomes between groups (ICP vs. GCP) subsequently diminished overtime. These findings indicate that using ICP for calving delays exposure to MAP in calves and provides an effective strategy for reducing peripartum MAP transmission risks in herds attempting to limit the impact of paratuberculosis.