Manuka honey chelates iron and impacts iron regulation in key bacterial pathogens.

AIM: The aim of this study was to test the hypothesis that Manuka honey (MH) chelates iron and promotes an iron-limiting environment, which contributes to its antimicrobial activity. METHODS AND RESULTS: Employing a ferrozine-based assay, we observed that MH is an iron chelator that depletes iron from solution. Siderophores are small molecules that bind ferric iron (III) with high affinity and their levels are upregulated by bacteria under iron-limiting conditions. We demonstrated by quantitating siderophore production that Escherichia coli and Pseudomonas aeruginosa treated with MH sub-minimum inhibitory concentrations (sub-MIC) experience an iron-limiting environment and increase siderophore production. In addition, supplementation with ferrous iron (II) significantly increased growth of E. coli, Staphylococcus aureus and P. aeruginosa cultured at their MH MIC above that observed in nonsupplemented controls. By contrast, supplementation with ferric iron (III) significantly increased growth for only E. coli and P. aeruginosa, above their nonsupplemented controls. CONCLUSIONS: Manuka honey chelates iron, thereby generating an iron-limiting environment for E. coli and P. aeruginosa, and to a lesser extent S. aureus, which contributes to its antimicrobial properties. SIGNIFICANCE AND IMPACT OF THE STUDY: Our work demonstrates that MH-induced iron chelation is an antimicrobial mechanism that differentially impacts the bacterial species tested here. Iron chelation affects multiple diverse physiological processes in bacteria and would contribute to the lack of bacterial resistance to MH. Iron metabolism is tightly regulated; bacteria require this essential nutrient for survival, but in excess it is toxic. Additional exploration of MH's iron chelation mechanism will facilitate its future use in mainstream medicine.

Energetic metabolism, milk production, and inflammatory response of transition dairy cows fed rumen-protected glucose.

Objectives were to evaluate the effects of rumen-protected glucose (RPG) supplementation on milk production, post-absorptive metabolism, and inflammatory biomarkers in transition dairy cows. Fifty-two multiparous cows were blocked by previous 305-d mature-equivalent milk (305ME) yield and randomly assigned to 1 of 2 iso-energetic and iso-nitrogenous treatments: (1) control diet (CON; n = 26) or (2) a diet containing RPG (pre-fresh 5.3% of dry matter and 6.0% of dry matter postpartum; n = 26). Cows received their respective dietary treatments from d -21 to 28 relative to calving, and dry matter intake was calculated daily during the same period. Weekly body weight, milk composition, and fecal pH were recorded until 28 d in milk (DIM), and milk yield was recorded through 105 DIM. Blood samples were collected on d -7, 3, 7, 14, and 28 relative to calving. Data were analyzed using repeated measures in the MIXED procedure (SAS Institute Inc., Cary, NC) with previous 305ME as a covariate. Fecal pH was similar between treatments and decreased (0.6 units) postpartum. Dry matter intake pre- and postpartum were unaffected by treatment, as was milk yield during the first 28 or 105 DIM. Milk fat, protein, and lactose concentration were similar for both treatments. Blood urea nitrogen and plasma glucose concentrations were unaffected by treatment; however, results showed increased concentration of circulating insulin (27%), lower nonesterified fatty acids (28%), and lower postpartum ß-hydroxybutyrate (24%) in RPG-fed cows. Overall, circulating lipopolysaccharide-binding protein and haptoglobin did not differ by treatment, but at 7 DIM, RPG-fed cows had decreased lipopolysaccharide-binding protein and haptoglobin concentrations (31 and 27%, respectively) compared with controls. Supplemental RPG improved some biomarkers of post-absorptive energetics and inflammation during the periparturient period, changes primarily characterized by increased insulin and decreased nonesterified fatty acids concentrations, with a concomitant reduction in acute phase proteins without changing milk production and composition.

Purinergic system in psychiatric diseases.

Psychiatric disorders are debilitating diseases, affecting >80 million people worldwide. There are no causal cures for psychiatric disorders and available therapies only treat the symptoms. The etiology of psychiatric disorders is unknown, although it has been speculated to be a combination of environmental, stress and genetic factors. One of the neurotransmitter systems implicated in the biology of psychiatric disorders is the purinergic system. In this review, we performed a comprehensive search of the literature about the role and function of the purinergic system in the development and predisposition to psychiatric disorders, with a focus on depression, schizophrenia, bipolar disorder, autism, anxiety and attention deficit/hyperactivity disorder. We also describe how therapeutics used for psychiatric disorders act on the purinergic system.

Modeling the trade-off between diet costs and methane emissions: A goal programming approach.

Enteric methane emission is a major greenhouse gas from livestock production systems worldwide. Dietary manipulation may be an effective emission-reduction tool; however, the associated costs may preclude its use as a mitigation strategy. Several studies have identified dietary manipulation strategies for the mitigation of emissions, but studies examining the costs of reducing methane by manipulating diets are scarce. Furthermore, the trade-off between increase in dietary costs and reduction in methane emissions has only been determined for a limited number of production scenarios. The objective of this study was to develop an optimization framework for the joint minimization of dietary costs and methane emissions based on the identification of a set of feasible solutions for various levels of trade-off between emissions and costs. Such a set of solutions was created by the specification of a systematic grid of goal programming weights, enabling the decision maker to choose the solution that achieves the desired trade-off level. Moreover, the model enables the calculation of emission-mitigation costs imputing a trading value for methane emissions. Emission imputed costs can be used in emission-unit trading schemes, such as cap-and-trade policy designs. An application of the model using data from lactating cows from dairies in the California Central Valley is presented to illustrate the use of model-generated results in the identification of optimal diets when reducing emissions. The optimization framework is flexible and can be adapted to jointly minimize diet costs and other potential environmental impacts (e.g., nitrogen excretion). It is also flexible so that dietary costs, feed nutrient composition, and animal nutrient requirements can be altered to accommodate various production systems.

Mineral concentrations in diets, water, and milk and their value in estimating on-farm excretion of manure minerals in lactating dairy cows.

Thirty-nine commercial dairies in Merced County, California were enrolled in the present study to (1) compare lactating cow mineral intakes (via drinking water and total mixed ration) to the National Research Council (NRC) requirements, (2) evaluate the association between dietary concentrations of minerals with and without drinking water and adjusted for mineral concentrations in milk, and (3) compare 4 different methods to estimate excretion of minerals using either assays or estimations of milk mineral outputs and total daily mineral intake per cow with or without minerals coming from drinking water. Dairies were selected to represent a range of herd milk yields and a range of water mineral contents. Samples of total mixed ration, drinking water, and bulk tank milk were taken on 2 different days, 3 to 7d apart in each farm. Across-farm medians and percentile distributions were used to analyze results. The herd median milk yield interquartile ranged (10th to 90th percentile) from less than 25 to more than 39 kg/d and the concentration of total solids in water interquartile ranged from less than 200 to more than 1,490 mg/L. Including drinking water minerals in the diets increased dietary concentrations by <4% for all minerals except for Na and Cl, which increased by 9.3 and 6.5%, respectively. Concentrations of P and K in milk were essentially the same as the NRC value to estimate lactation requirements. However, NRC milk values of Ca, Cl, and Zn were 10 to 20% greater than dairy farm values; and Na, Cu, Fe, and Mn were no less than 36% below NRC values. Estimated excretion of minerals via manure varied substantially across farms. Farms in the 10th percentile did have 2 to 3 times less estimated mineral excretions than those in the 90th percentile (depending on the mineral). Although including water minerals increased excretion of most minerals, the actual median effect of Ca, Mg, S, Cu, Fe, and Mn was less than 5%, and about 8% for Na and Cl. Replacing assayed concentrations of minerals in milk with NRC constants resulted in reduced estimated excretion of Ca, Na, Cu, Fe, and Zn, but median differences were <5% except for Na which was 7.5%. Results indicate that not including mineral intake via drinking water and not using assayed concentrations of milk minerals lead to errors in estimation manure excretion of minerals (e.g., Ca, Na, Cl, and S).

Evaluation of a biological risk management tool on large western United States dairies.

Critical to changing biosecurity practices on the farm is an individual assessment of those practices contributing to disease transmission. The purpose of this project was to assess, implement, and refine a biological risk management survey for use on large western United States dairy farms. Assessment tools developed by Iowa State University Center for Food Security and Public Health (Ames, IA) were refined using a focus group process and by testing them on 40 dairy herds in California. Each question was evaluated using standard criteria and producer responses. Some survey questions required refinement for clarity and others were considered unnecessary. New questions were added based on a biosecurity literature review, resulting in a new set of questions that can be used by extension educators and food animal veterinarians to help identify disease risk areas and educate dairy producers.

Precipitation and temperature effects on mortality and lactation parameters of dairy cattle in California.

Data from 3 commercial rendering companies located in different regions of California were analyzed from September 2003 through August 2005 to examine the relationship of dairy calf and cow mortality to monthly average daily temperature and total monthly precipitation respectively. Yearly average mortality varied between rendering regions from 2.1 to 8.1% for mature cows. The relationship between cow and calf monthly mortality and monthly average daily temperature was U-shaped. Overall, months with average daily temperatures less than 14 and greater than 24 degrees C showed substantial increases in both calf and cow mortality with calf mortality being more sensitive to changes in these temperature ranges than cow mortality. Temperature changes were reflected in a 2-fold difference between the minimum and maximum mortality in cows and calves. Precipitation showed a weak effect with calf mortality and no effect with cow mortality. Data from Dairy Herd Improvement Association were used from 112 California herds tested over a 24-mo period to examine the relationship of milk production and quality with monthly average daily temperature and monthly precipitation. Somatic cell count and percent milk fat were either weakly or not associated with monthly average daily temperature and total monthly precipitation. However, total monthly precipitation was negatively associated with test day milk per milking cow regardless of the dairy's geographical location. Housing-specific associations for test day milk per milking cow were greater for total monthly precipitation than monthly average daily temperature, with the strongest negative association seen for dairies that do not provide shelter for cows. This suggests that providing suitable housing for lactating dairy cattle may ameliorate the precipitation-associated decrease in test day milk per milking cow.

Yeast Dam1p is required to maintain spindle integrity during mitosis and interacts with the Mps1p kinase.

We have identified a mutant allele of the DAM1 gene in a screen for mutations that are lethal in combination with the mps1-1 mutation. MPS1 encodes an essential protein kinase that is required for duplication of the spindle pole body and for the spindle assembly checkpoint. Mutations in six different genes were found to be lethal in combination with mps1-1, of which only DAM1 was novel. The remaining genes encode a checkpoint protein, Bub1p, and four chaperone proteins, Sti1p, Hsc82p, Cdc37p, and Ydj1p. DAM1 is an essential gene that encodes a protein recently described as a member of a microtubule binding complex. We report here that cells harboring the dam1-1 mutation fail to maintain spindle integrity during anaphase at the restrictive temperature. Consistent with this phenotype, DAM1 displays genetic interactions with STU1, CIN8, and KAR3, genes encoding proteins involved in spindle function. We have observed that a Dam1p-Myc fusion protein expressed at endogenous levels and localized by immunofluorescence microscopy, appears to be evenly distributed along short mitotic spindles but is found at the spindle poles at later times in mitosis.

The effect of energy supplementation on nitrogen utilization in lactating dairy cows fed grass silage diets.

An experiment was conducted to examine the effect that various isoenergetic diets, containing different quantities of soluble carbohydrate and fiber and different types of starch, have on nitrogen (N) balances. Six lactating dairy cows in early to midlactation consuming grass silage diets with not less than 600 g/kg total DMI as forage were used in the experiment. Four concentrates were prepared that had higher amounts of either fiber, soluble sugars, corn (low degradable starch source), or barley (high degradable starch source). Overall N utilization by the cows was poor, rarely exceeding 0.30 g milk N/g of dietary N intake. Fecal N outputs accounted for more than half of total N excreted in all treatments except for diets supplemented with high degradable starch, in which urinary N excretion was significantly higher compared with the other treatments. Milk yield was unaffected by concentrate type, averaging 19.9 kg/d, but milk protein content decreased from 32.9 for starch-based diets to 30.9 and 30.0 g/kg for the soluble sugar- and fiber-based diets, respectively. The efficiency of N utilization improved in the low degradable starch treatment, which had lower N excretion (65%) and higher protein concentration in milk. Furthermore, feeding cows corn-based concentrates reduced urinary N excretion by almost 30% compared with barley-based concentrates; therefore, feeding corn-based diets is recommended for the reduction of nitrogen pollution in lactating dairy cows.

The effect of protein supplementation on nitrogen utilization in lactating dairy cows fed grass silage diets.

The study set out to examine the effects of supplementing grass silage with various levels of protein concentration and degradability on dietary nitrogen (N) excretion in lactating dairy cows consuming at least 60% forage. Six Holstein/Friesian cows in early to midlactation were offered six diets comprising two levels of crude protein (210 and 290 g/kg DM) and three levels of protein degradability in the concentrate achieved using different amounts of untreated or formaldehyde-treated soybean meal. Despite a difference of almost 100 g/d in N intake, apparent fecal and milk N outputs were not significantly affected. Protein degradability also had no effect on N outputs in feces and milk. However, there was a major effect of both level and degradability of CP on urinary N output. Moreover, an interaction between level and degradability of CP was detected, such that the rate at which urinary N increases with increasing CP degradability was higher on the high-CP than on the low-CP diet. A low level of protein (150 g/kg DM in the diet) and medium to low rumen-degradable protein supplements provided a significant reduction in N excretion without compromising lactational performance (mean 24.8 kg/d), in terms of both milk yield and composition. This study also demonstrated that a high efficiency of N utilization could be achieved on low-CP diets (supplying less than 400 g N/d), with feces being the main route of N excretion, whereas an exponential excretion of urinary N was observed as N intake exceeded 400 g N/d.

Monensin for lactating dairy cows grazing mixed-alfalfa pasture and supplemented with partial mixed ration.

The effect of monensin on milk production was evaluated in 58 lactating Holstein cows (48 multiparous; 10 primiparous) grazing a mixed-alfalfa pasture and supplemented with a partial mixed ration in a completely randomized design with repeated measurements. Cows were paired by calving date, lactation number, previous lactation milk production, body weight, and body condition score and were assigned to one of 2 treatments: control or monensin. Cows on the monensin treatment received 2 monensin controlled-release capsules (335 mg/d for 90 d), one 30 d before the expecting calving date and the other 60 d after calving. Short-term (0 to 150 d in milk) and long-term (305-d adjusted lactation) effects of monensin were evaluated. Pasture (measured by difference between pre- and postgrazing pasture mass), supplements, and total dry matter intake did not differ between treatments and averaged 8.7, 14.1, and 22.9 kg/d, respectively. In the short-term, monensin increased milk production (27.7 vs. 26.6 kg/d) and milk protein yield (0.890 vs. 0.860 kg/d); milk fat yield was not affected (0.959 kg/d). Monensin decreased milk fat content (3.51 vs. 3.60%) with no changes in milk protein content (3.25%). In the long term, milk production and milk protein yield were also increased by monensin: 214 and 7 kg, respectively. Monensin reduced the loss of body condition score and increased percentage of pregnancy at first service (44.8 vs. 20.7%). Monensin improves production and reproduction performance of dairy cows grazing a mixed-alfalfa pasture and supplemented with a partial mixed ration.

Brain SPECT imaging in children and adolescents with obsessive-compulsive disorder.

OBJECTIVE: To evaluate the patterns of regional cerebral blood flow (rCBF) in cortical and subcortical regions by Brain SPECT imaging, in children and adolescents with obsessive-compulsive disorder (OCD) before and after treatment. METHOD: Fourteen OCD patients (6 to 17 years old) underwent brain SPECT; ten of those subjects were reexamined after successful treatment. rCBF ratios were correlated with clinical parameters on the 14 patients in symptomatic state, and we compared rCBF ratios of the ten patients before and after treatment. RESULTS: There was no statistically significant difference in average ratios of rCBF before and after treatment. There were significant clinical correlations between current age and age of onset of OCD and rCBF in the bilateral superior frontal, and bilateral parietal cortical regions. CONCLUSIONS: Further investigations on abnormal neurodevelopment of cortical-subcortical circuits possibly involved in symptomatology of paediatric OCD are warranted.

The Arabidopsis-accelerated cell death gene ACD2 encodes red chlorophyll catabolite reductase and suppresses the spread of disease symptoms.

accelerated cell death 2 (acd2) mutants of Arabidopsis have spontaneous spreading cell death lesions and constitutive activation of defenses in the absence of pathogen infection. Lesion formation in acd2 plants can be triggered by the bacterial toxin coronatine through a light-dependent process. Coronatine-triggered and spontaneous lesion spreading in acd2 plants also requires protein translation, indicating that cell death occurs by an active process. We have cloned the ACD2 gene; its predicted product shows significant and extensive similarity to red chlorophyll catabolite reductase, which catalyzes one step in the breakdown of the porphyrin component of chlorophyll [Wüthrich, K. L., Bovet, L., Hunziger, P. E., Donnison, I. S. & Hörtensteiner, S. (2000) Plant J. 21, 189-198]. Consistent with this, ACD2 protein contains a predicted chloroplast transit peptide, is processed in vivo, and purifies with the chloroplast fraction in subcellular fractionation experiments. At some stages of development, ACD2 protein also purifies with the mitochondrial fraction. We hypothesize that cell death in acd2 plants is caused by the accumulation of chlorophyll breakdown products. Such catabolites might be specific triggers for cell death or they might induce cellular damage through their ability to absorb light and emit electrons that generate free radicals. In response to infection by Pseudomonas syringae, transgenic plants expressing excess ACD2 protein show reduced disease symptoms but not reduced growth of bacteria. Thus, breakdown products of chlorophyll may act to amplify the symptoms of disease, including cell death and yellowing. We suggest that economically important plants overexpressing ACD2 might also show increased tolerance to pathogens and might be useful for increasing crop yields.

Effects of management practices prior to and during ensiling and concentrate type on nitrogen utilization in dairy cows.

Three experiments with a total of 18 multiparous Holstein-Friesian dairy cows in early to mid lactation were used in incomplete changeover designs with four periods consisting of 4 wk each. The objectives of the experiments were to investigate the effects of 1) grass management practices prior to ensiling and 2) concentrate energy sources on animal performance and N utilization and suggest ways in which the annual N excretion can be reduced. In the first experiment, four silages were prepared as early-cut material from grass that was fertilized with either 75 or 150 kg of N/ha. Two other silages were prepared from the latter but harvested 2 wk later. Within each silage type, both direct cut and wilted silages were produced. In the second experiment, starch-based and fiber-based concentrates were offered with the three wilted silages used in the previous experiment. In the third experiment, barley-based and corn-based concentrates were offered with three silages that had been treated with different additives at the time of ensiling. Neither dry matter intake nor milk yield were significantly affected by either silage or concentrate type. Milk protein content and yield, however, were higher in silages made from medium fertilized grass and also increased with increased starch content of the concentrate. Grass management and diet affected the amount and form of N excreted. Use of nitrogen fertilizer at lower than 75 kg of N/ha and feeding less degradable starch concentrate is recommended for mitigation of N pollution, especially in the form of urine N by dairy cows in intensive farming.

Effects of feeding rations with genetically modified whole cottonseed to lactating Holstein cows.

Two experiments were conducted to evaluate dry matter intake (DMI), milk yield, and milk composition from feeding rations that contained different sources of genetically modified whole cottonseed to Argentinean Holstein dairy cows. Twenty-four lactating multiparous Argentinean Holstein dairy cows were used in 2 experiments with a replicated 4 x 4 Latin square design, with cows averaging 565 kg body weight and 53 d in milk at the beginning of the experiments. Treatments in Experiment 1 were: Bollgard cotton containing the cry1Ac gene, Bollgard II cotton containing cry1Ac and cry2Ab genes, Roundup Ready cotton containing the cp4 epsps gene, and a control nongenetically modified but genetically similar cottonseed. In Experiment 2, two commercial sources, a parental control line, and the transgenic cotton containing both cry1Ac and cp4 epsps genes were used as treatments. All cows received the same total mixed ration but with different whole cottonseed sources. Cottonseed was included to provide 2.50 kg per cow daily (dry matter [DM] basis) or about 10% of the total diet DM. The ingredient composition of the total mixed ration was 32% alfalfa hay, 28% corn silage, 22% corn grain, 17% soybean meal, and 2% minerals and vitamins. In addition, genomic DNA was extracted from a subset of milk samples and analyzed by polymerase chain reaction followed by Southern blot hybridization for small fragments of the cry1Ac transgene and an endogenous cotton gene, acp1. No sample was positive for transgenic or plant DNA fragments at the limits of detection for the assays following detailed data evaluation criteria. The DMI, milk yield, milk composition, body weight, and body condition score did not differ among treatments. Cottonseed from genetically modified varieties used in these studies yielded similar performance in lactating dairy cows when compared to non-transgenic control and reference cottonseed.

Características clínicas de transtorno obsessivo-compulsivo em crianças e adolescentes / Clinical features of obsessive-compulsive disorder in children and adolescents

OBJETIVO: Descrever o quadro clínico, evoluçäo, comorbidade, subtipo de transtorno obsessivo-compulsivo (TOC) e incidencia de patologia familiar em uma amostra clínica de crianças e adolescentes com TOC.CASUISTICA E METODOS: Foram avaliados prospectivamente 40 pacientes menores de 18 anos, do Serviço de Psiquiatria da Infancia e Adolescencia(SEPIA), com TOC, no periodo de junho de 1995 até agosto de 1998, através da entrevista K-SADs, versäo E -IV e escala de sintomas Y-BOCS. Dados de incidencia de patologia familiar foram obtidos por anamnese clínica. Dados de evoluçäo säo referentes ao período em que estiveram em seguimento no SEPIA e, retrospectivamente, desde o início dos sintomas pela história clínica. RESULTADOS: A maioria dos pacientes era do sexo masculino (77,5 por cento), com idade média de início de 10,04 anos. Houve predomínio de subtipo misto de obsessöes e compulsöes, com tipo de sintomas em geral múltiplos e mutáveis ao longo do tempo. Comorbidade foi muito freqüentes nesta amostra (10 por cento). Houve 22,5 por cento de familiares com TOC ou SOC (sintomas obsessivo-compulsivos; subclínico). A evoluçäo foi na maioria crônica flutuante. CONCLUSOES: Trata-se de uma amostra clínica em que os pacientes foram selecionados de uma populaçäo rica em casos psiquiatricos gerais mais graves, por ser um centro de referencia para este atendimento. Portanto há vies quanto ao grau de acometimento e talvez isto tenha influenciado para diferenças em relaçäo a dados de literatura sobre TOC infantil