Evolutionary discrimination of mammalian conserved non-genic sequences (CNGs)

Analysis of the human and mouse genomes identified an abundance of conserved non-genic sequences (CNGs). The significance and evolutionary depth of their conservation remain unanswered. We have quantified levels and patterns of conservation of 191 CNGs of human chromosome 21 in 14 mammalian species. We found that CNGs are significantly more conserved than protein-coding genes and noncoding RNAS (ncRNAs) within the mammalian class from primates to monotremes to marsupials. The pattern of substitutions in CNGs differed from that seen in protein-coding and ncRNA genes and resembled that of protein-binding regions. About 0.3% to 1% of the human genome corresponds to a previously unknown class of extremely constrained CNGs shared among mammals.     

Seasonal Variation of Phenolic Antioxidant-mediated α-glucosidase Inhibition of <Emphasis Type="Italic">Ascophyllum nodosum</Emphasis>

Ascophyllum nodosum is a brown seaweed that grows abundantly in the US Northeast coastal region. This study examined the seasonal variation of A. nodosum in phenolic contents and subsequent antioxidant, α-glucosidase and α-amylase inhibitory activities. A. nodosum was harvested monthly and extracted in hot water and the resulting extracts were spray-dried. The results indicate a clear seasonal variation in terms of phenolic content, with June and July being the highest (36.4 and 37 mg/g, respectively) and May the lowest (21.8 mg/g). The antioxidant activities, in terms of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, correlated with the phenolic contents observed (r = 0.81), with the month of July being the highest (58%) and April the lowest (26%). Similarly in terms of Trolox equivalent, July had the highest activity (15.53 μM) and April and May the lowest (8.40 and 8.27 μM, respectively). α-glucosidase inhibitory activity exhibited a pattern similar to the phenolic contents observed with July having the highest inhibitory activity (IC₇₀ 2.23 μg) and April the lowest (IC₇₀ 26.13 μg), resulting in an inverse correlation between IC₇₀ values and total phenolic content (r = −0.89). Such seasonal variation is believed to be caused by temperature-related stress considering that A. nodosum is a cold water species.     

Determination of baseline susceptibility of European populations of Tuta absoluta (Meyrick) to indoxacarb and chlorantraniliprole using a novel dip bioassay method

BACKGROUND: Tuta absoluta(Meyrick) is one of the most serious pests of tomato recently introduced in the Mediterranean region. A novel bioassay method designed for the accurate determination of insecticide toxicity on T. absoluta (IRAC method No. 022) was validated by three different laboratories [Greece (NAGREF), Italy (UC) and Spain (UPCT)] on European populations. RESULTS: The insecticides indoxacarb and chlorantraniliprole were used as reference products. The IRAC leaf dip method is easy to perform, producing repeatable, homogeneous responses. LC₅₀ values for indoxacarb ranged between 1.8 and 17.9 mg L^?1 (NAGREF), 0.93 and 10.8 mg L^?1 (UC) and 0.20 and 0.70 mg L^?1 (UPCT), resulting in a tenfold, 12‐fold and fourfold difference between the least and most susceptible populations at each laboratory respectively. For chlorantraniliprole, LC₅₀ values ranged between 0.10 and 0.56 mg L^?1 (NAGREF), 0.23 and 1.34 mg L^?1 (UC) and 0.04 and 0.24 mg L^?1 (UPCT), resulting in a sixfold difference in all three cases. Overall, UPCT reported lower mean LC₅₀ to indoxacarb, while UC reported higher LC₅₀ to chlorantraniliprole. CONCLUSIONS: The new bioassay is reliable, providing a useful tool in the design of IRM strategies. Within each country/lab, the variability observed in the results for both indoxacarb and chlorantraniliprole can be attributed to natural variation. Future research is necessary to determine the extent to which it is possible to compare results among laboratories. Copyright © 2012 Society of Chemical Industry     

Recovery of Bioactive Peptides and Omega-3 Fatty Acids-Containing Phospholipids from Squid Processing By-Product Hydrolysate

ABSTRACT

The present study examined whether bioactive peptides and omega-3 fatty acids-containing phopholipids could be recovered from squid processing by-product (SPB) hydrolysate. The hydrolysate was produced at 55°C for varying times with endogenous proteases and centrifuged to yield peptides-containing supernatant and phospholipids-containing precipitate. The supernatant showed angiotensin I-converting enzyme (ACE) inhibitory activity with half maximal inhibitory concentration (IC₅₀) values decreasing with hydrolysis time from 2.11 mg (0 h) to 1.71 mg (1 h), 1.38 mg (1.5 h), and 1.34 mg (2 h). Two-hour squid hydrolysate was further fractionated to isolate the most active fraction whose molecular weight was found to be below 10 kDa with IC₅₀ of 0.32 mg. The phospholipids-containing precipitate (45.6 g/100 g oil) was analyzed for a fatty acid profile, with the levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) being 16.86 and 29.24 g/100 g oil, respectively. The ACE inhibition by hydrolysate peptides and omega-3 fatty acid recovery support the nutraceutical potential of the SPB hydrolysate.     

Q fever

Q fever is a zoonotic disease caused by the ubiquitous pathogen Coxiella burnetii responsible for acute and chronic clinical manifestations. Farm animals and pets are the main reservoirs of infection, and transmission to human beings is mainly accomplished through inhalation of contaminated aerosols. This illness is associated with a wide clinical spectrum, from asymptomatic or mildly symptomatic seroconversion to fatal disease. In humans Q fever can manifest as an acute disease (mainly as a self-limited febrile illness, pneumonia, or hepatitis) or as a chronic disease (mainly endocarditis), especially in patients with previous valvulopathy and to a lesser extent in immunocompromised hosts and in pregnant women. In contrast in animals, Q fever is in most cases, strikingly asymptomatic. The definite diagnosis of Q fever is made based on a significant increase in serum antibody titers, the determination of which often requires considerable time, and therefore patients must be monitored for a certain period. The treatment is effective and well tolerated, but must be adapted to the acute or chronic pattern with the tetracyclines to be considered the mainstay of antibiotic therapy. Several actions have been proposed to prevent and reduce the animal and environmental contamination. Vaccination of animals in infected flocks, as well as in uninfected ones close to them, with an efficient vaccine can prevent abortions and shedding of the bacteria.     

Evaluation of ornithine carbamoyl transferase and other serum and liver-derived analytes in diagnosis of fatty liver and postsurgical outcome of left-displaced abomasum in dairy cows

OBJECTIVE: To evaluate postsurgical outcome in dairy cows with left-displaced abomasum (LDA) with regard to severity of fatty liver and assess the usefulness of preoperative determination of serum ornithine carbamoyl transferase (OCT) activity, bile acids concentration, and other variables for evaluating liver function during the postsurgical convalescence period. DESIGN: Prospective study. ANIMALS: 68 Holstein cows. PROCEDURES: Blood and liver biopsy specimens were obtained during standing LDA surgery. Liver tissue was examined histologically and classified by severity of fatty change. Serum activities of liver-derived enzymes and concentrations of total lipids, triglycerides, bile acids, glucose, beta-hydroxybutyric acid, bilirubin, and nonesterified fatty acids were determined. RESULTS: Most cows with LDA and cows with severe fatty liver were detected within the first month after calving. Postsurgical outcome was related to severity of fatty liver. All cows that died had severe fatty liver. Serum activities of OCT, aspartate aminotransferase, and glutamate dehydrogenase and serum total bilirubin concentration were sensitive indicators of fatty liver. Serum bile acids concentration was not an accurate indicator of fatty liver. CONCLUSIONS AND CLINICAL RELEVANCE: Postsurgical outcome of cows undergoing surgery to correct LDA was related to fatty liver severity. Assessment of serum activities of OCT, aspartate aminotransferase, and glutamate dehydrogenase and serum total bilirubin concentration is recommended for diagnosis of fatty liver in dairy cows with LDA, whereas determination of bile acids concentration is not. The strong correlation between OCT activity and degree of hepatocellular damage supports use of this enzyme for assessing severity of fatty liver and predicting postsurgical outcome in cows with LDA.     

Genetic variation in pea seed globulin composition

A quantitative characterization of seeds from 59 pea (Pisum sativum L.) lines and relative taxa with various external characteristics and wide geographical origin was performed to explore the genetic variation of pea concerning its starch and protein contents and globulin composition. Pea lines, which produce round, wrinkled, flat, and round-dimpled seeds, have starch as the major reserve, with an average content of 46%. Protein content varied from 13.7 to 30.7% of the seed dry matter, with an overall average of 22.3%. Densitometric quantification of the individual globulins (legumin, vicilin, convicilin, and globulin-related proteins) based on SDS-PAGE gels showed no lines lacking any particular globulin. Among the lines tested, variation was shown in both their total globulins content and their globulin composition. The total globulin content ranged from 49.2 to 81.8% of the total pea protein extract (TPPE). Legumin content varied between 5.9 and 24.5% of the TPPE. Vicilin was the most abundant protein of pea, and its content varied between 26.3 and 52.0% of the TPPE. Both processed and nonprocessed vicilins occurred. The processed vicilin was the predominant one, with values between 17.8 and 40.8%, whereas the nonprocessed ones constituted between 3.1 and 13.5% of the TPPE. Convicilin was the least abundant globulin, and its content ranged from 3.9 to 8.3%. Finally, the globulin-related proteins were present in amounts ranging from 2.8 to 17.3%. They were less abundant in comparison with legumin and vicilin, but they showed the largest relative variation of the four globulin classes. Correlations between the different external characteristics and globulin composition were determined. Comparison with soybean showed that pea lines show more variety in the abundance of globulin proteins, enabling a wider range of food application.     

Microwave-assisted extraction (MAE)-acid hydrolysis of dithiocarbamates for trace analysis in tobacco and peaches

A simple and rapid method is presented for the analysis of residues of ethylenebis(dithiocarbamate) (maneb, zineb, and mancozeb) and N,N-dimethyldithiocarbamate (thiram and ziram) fungicides in dry tobacco leaves and peaches. Residues are extracted and hydrolyzed to CS(2) in a single step by use of microwave energy in a closed-vessel system while the evolved CS(2) trapped in a layer of iso-octane overlaying the reaction mixture is taken for gas chromatographic-flame photometric analysis. This combined extraction-hydrolysis step is carried out in 10 and 15 min for sets of 12 samples of tobacco and peach matrices, respectively. Total sample preparation time for GC analysis is 40 min. The limits of detection (LOD) are 0.005 mg/kg for thiram and ziram on peaches and 0.1 mg/kg for maneb, zineb, and mancozeb on tobacco. The respective LOD and limit of quantification (LOQ) levels in CS(2) equivalents are 0.003 and 0.006 mg/kg on peaches and 0.04 and 0.2 mg/kg on tobacco, respectively. Recoveries in the 0.01-60 mg/kg fortification range are 80-100% with respective relative standard deviations <20%. The method was used for the analysis of >3000 commercial tobacco samples including also different marketed cigarette brands.     

Claw histopathology and parasitic load in natural cases of canine leishmaniosis associated with Leishmania infantum

Histological lesions and the presence of Leishmania spp. amastigotes in claw tissues were investigated in 40 dogs with leishmaniosis, with (16/40--group A) or without (24/40--group B) generalized onychogryphosis. Following euthanasia, the entire third phalanx with intact claw was amputated, formalin fixed, decalcified in a formic acid solution, embedded in paraffin, sectioned longitudinally and stained with haematoxylin and eosin, and acid orcein-Giemsa. Nested polymerase chain reaction (PCR) was used for the detection of Leishmania amastigotes. Lichenoid mononuclear infiltration (all dogs in group A, 21 of 24 dogs in group B), basal keratinocyte vacuolation (nine of 16 dogs in group A, 15 of 24 dogs in group B) and dermoepidermal clefting (13 of 16 dogs in group A, 18 of 24 dogs in group B) were the most prominent histopathological findings. There was no difference in the frequency and severity of these lesions between the two groups. Leishmania amastigotes could not be visualized in the dermis of any of the H&E sections, but their presence was demonstrated by nested PCR in three of 16 dogs in group A and two of 24 dogs in group B. However, the frequency of positive nested PCRs was not significantly different between the two groups. In conclusion, claw histopathology in symptomatic dogs with leishmaniosis, either with or without onychogryphosis is mainly characterized by mononuclear lichenoid dermatitis with or without interface dermatitis and dermoepidermal clefting, and is not accompanied by substantial local parasitism.     

Novel methodologies in screening and selecting olive varieties and root-stocks for resistance to <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

An innovative inoculation process, involving the drilling of a trunk hole in 3 year-old olive trees and injecting a dense conidial suspension of Verticillium dahliae, was developed to study differentiation in foliar symptom expression between olive cultivars tolerant or susceptible to the pathogen. It was demonstrated that V. dahliae conidia could be translocated and colonize the xylem at the same distance above and below the point of trunk injection in both cultivars. However, the pathogen could be subsequently isolated at statistically significant percentages in susceptible cv. Amphissis compared to the tolerant cv. Kalamon, indicating operation of resistance mechanisms in the vascular phase of the disease. Consequently symptom development in the susceptible cultivar was at least sixfold more intensive compared to the tolerant cultivar, 6–11 months after trunk inoculation. Perennial olive orchard experiments, aimed at selecting Verticillium-resistant root-stocks, were conducted by applying the novel method in 2–3 year-old root-stock suckers of Amphissis olive trees and in the tolerant cvs Lianolia of Corfu and Koroneiki. It was indicated that potentially resistant root-stocks could be obtained following the trunk drilling technique. Resistance differentiation between cvs Amphissis and Kalamon was further verified through root inoculation by various V. dahliae microsclerotial concentrations and demonstrated that the trunk drilling inoculation procedure is equally efficient in resistance evaluation of olives to Verticillium wilt. The trunk inoculation procedure could be useful in selecting and screening root-stocks for resistance to V. dahliae and other vascular pathogens and could elucidate resistance mechanisms in woody plants against vascular wilt diseases.