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Characterization of Novel Luteoviruses in Canadian Highbush Blueberries Using High-Throughput Sequencing.

The Fraser Valley of British Columbia, Canada is among the top ten blueberry producing regions globally. Viral diseases are established in the region and significantly reduce average yields. While testing for two viruses is routine, characterization of all the viruses present in the region is incomplete. We used high-throughput sequencing to obtain an unbiased overview of RNA viruses present in 97 plants collected across the region. In addition to known viruses, we identified four luteoviruses previously unidentified in the region. Two of them matched the blueberry virus L (BlVL) and blueberry virus M (BlVM). recently found in the USA, while the third constitutes a new major variant of BlVM (BlVM-2), and the fourth a new luteovirus, which we named blueberry virus N (BlVN). The genome sequences were ~5 kbp long and contained four open-reading frames similar to other luteoviruses. PCR screening revealed that these luteoviruses are widespread in the region, and that plants typically harbour more than one of these luteoviruses. While luteoviruses are typically vectored by aphids, they were also present in nursery stock, indicating that spread also occurs via vegetative propagation.

High-Throughput Nucleotide Sequencing

Determination of carbendazim in blueberries by reversed-phase high-performance liquid chromatography.

A fluorescent reversed-phase high-performance liquid chromatographic method was developed for the analysis of carbendazim in blueberries. Recoveries of fortified blueberries at 27 and 810 ng/g were more than adequate with good precision. Forty commercial blueberry samples were analyzed and the amount of carbendazim ranged from none detected (detection limit of 15 ng/g) to 155 ng/g. Confirmation of carbendazim in the blueberry samples was made by enzyme immunoassay and UV photodiode array.

Benzimidazoles

Supplementations of docosahexaenoic acid and blueberry suppress a high-fat breakfast-induced postprandial inflammation but only docosahexaenoic acid improves endothelial function in healthy adults: a randomized, double-blind, placebo-controlled crossover intervention study.

Blueberries and n - 3 polyunsaturated fatty acids each can provide protection against inflammation and cardiometabolic disorders. However, the underlying mechanisms are not fully understood. We hypothesized that blueberry and docosahexaenoic acid (DHA) can suppress high fat (HF) meal-induced postprandial inflammation and improve endothelial function. Sixty-two healthy participants (age: 26.8 &#xb1; 1.2 y; BMI: 22.1 &#xb1; 1.2 kg/m&#xb2;) consumed an isoenergetic breakfast (850 kcal) containing 34.7 g mostly animal fat (36% kcal), 25.3 g protein, and 111 g carbohydrate, with or without either 42.2 g blueberry powder (BBP) or 1.76 g DHA in a randomized, double-blind, placebo-controlled crossover intervention study. Blood samples were collected before and 1 h, 3 h and 6 h after breakfast. Monocyte activation, proinflammatory gene expression, cytokine production and endothelial function were assessed. Compared with the placebo control, DHA supplementation suppressed the HF breakfast-induced: expression of IL-1&#x3b2; by 21.6% (P < .01) and prostaglandin-endoperoxide synthase 2 (PTGS2, i.e., cyclooxygenase 2) by 22.8% (P < .01) at 6 h; plasma IL-1&#x3b2; production by 40.1 to 49.8% (P < .01) at 1-6 h; lipoprotein lipase (LPL)-treated blood IL-1&#x3b2; production by 40.9% (P < .0001) at 6 h; and total cholesterol/HDL cholesterol ratio by 2.2% (P < .01) at 3 h and 3.5% (P < .0001) at 6 h. BBP supplementation suppressed LPL-treated blood IL-1&#x3b2; production by 23.1% (P < .05) at 6 h. BBP and DHA also induced postprandial increases in reactive hyperemia index (RHI) scores relative to the fasting baselines, with DHA producing a 13.3% increase compared with placebo at 6 h (P < .05). In conclusion, supplementation with BBP or DHA suppressed the HF meal-induced postprandial inflammation but only DHA improved postprandial endothelial function. This study was registered at clinicaltrails.gov (NCT02472171).

Blueberry

Bacteriophages Control Epiphytic Pseudomonas syringae Populations in Highbush Blueberry Leaves.

The Pseudomonas syringae complex (Psc) is a group of globally distributed phytopathogens responsible for substantial agricultural losses. Although bacteriophage-based biocontrol has shown promise against Psc, no studies have examined phages targeting blueberry-tropic Psc lineages. Here, we isolated phages infecting Psc strains from diseased highbush blueberry (Vaccinium corymbosum), and evaluated their suitability for biocontrol using a multi-stage screening pipeline incorporating host-range analysis, comparative genomics, environmental stability testing, in&#xa0;vitro antibacterial efficacy assays and ex planta validation. Twelve of the isolated phages exhibited favourable host-range characteristics. Genomic analyses revealed substantial phylogenetic diversity among these candidates but simultaneously identified multiple clonal groups, reducing the collection to eight non-redundant phages spanning five distinct genera. Candidate phages generally retained infectivity under environmentally relevant conditions and exhibited heterogeneous but largely favourable stability profiles. Planktonic killing assays uncovered considerable variation in antibacterial efficacy, but phage performance appeared to be driven by infection compatibility and host-specific factors rather than properties intrinsic to individual phages. Notably, the jumbo phageCB10 emerged as a particularly promising candidate due to its strong antibacterial activity (median GRC&#x2009;=&#x2009;0.943), favourable environmental stability and unique genomic features. Cocktails containing the most effective candidates produced substantial and longitudinally sustained reductions in epiphytic colonization of detached blueberry leaves by Psc, exceeding five orders of magnitude at peak efficacy and demonstrating robust activity in a biologically relevant ex planta system. Importantly, in&#xa0;vitro antibacterial efficacy was predictive of performance in our ex planta model (r&#x2009;=&#x2009;0.67; p&#x2009;=&#x2009;0.0003), supporting the utility of tiered screening approaches for candidate selection. Taken together, these findings establish a framework for the systematic identification and evaluation of phages targeting Psc, and support the development of phage-based interventions for managing plant diseases.

Pseudomonas syringae

Transient blueberry muffin appearance of a newborn with congenital monoblastic leukemia.

A full-term male infant was born with skin findings suggesting a blueberry muffin appearance. Biopsy of a cutaneous nodule was consistent with monoblastic leukemia cutis, and bone marrow examination confirmed the diagnosis of leukemia. The infant has remained well 2 years after spontaneous resolution of the cutaneous eruption. Infiltrative neoplasms should be considered along with congenital infections and hematologic disorders in the differential diagnosis of a newborn with a blueberry muffin appearance.

Biopsy

A rapid GC method of monitoring Mesurol (4-(methylthio)-3,5-xylyl-N-methyl carbamate) and its sulfoxide and sulfone metabolites and their persistence in lowbush blueberries.

A facile analytical procedure was developed for determining Mesurol (4-(methylthio)-3,5-xylyl-N-methyl carbamate) and its oxidation products in blueberries. It involved blending with acetone, partition with chloroform and derivatization with trifluoroacetic anhydride and quantitation by gas chromatography/flame photometric detector (GC/FPD). The method showed good recoveries for Mesurol and its sulfoxide at the 0.1 ppm level and Mesurol sulfone at the 0.3 ppm level with a 25 g sample. It was applied to monitor levels of the insecticide and its oxidation products on field-treated blueberries. The sensitivity of the method may be increased 5-fold by the inclusion of a clean-up step. The optimal conditions for the detection of Mesurol TFA by the modified Bendix sulphur/phosphorus emission detector operating in the sulphur mode required an oxygen/hydrogen ratio of 0.38, for a column flow of 60 ml/min. The minimum detectable amounts of the TFA derivatives of Mesurol, its sulfoxide and sulfone were calculated as 1.3, 2.3 and 5.8 X 10(-11) g/sec, respectively.

Chromatography, Gas

[The phenolics of fruits. VI. The phenolics of currants, gooseberries and blueberries. Changes in phenolic acids and catechins during development of black currants (author's transl)].

Black, red, and white currants, gooseberries and cultivated blueberries contained only small amounts of catechins (total up to 30 mg per kg). Hydroxycinnamic acid derivatives, especially of caffeic and p-coumaric acid, occured mostly in higher concentrations. Furthermore, 4-hydroxygenzoic acid, protocatechuic acid and gallic acid were found after hydrolysis in currants and only protocatechuic acid in gooseberries in different levels. Salicyclic and gentisic acid were found in traces up to 2 mg per kg, occassionally vanillic acid, syringic acid or isoferulic acid. The concentrations of the examined phenolic acids in black currants related to the whole fruit (mg per fruit) increased, and related to fresh weight (mg per 1000 g) decreased during the growth of the fruits with the exception of protocatechuic acid, which appeared mostly in a last stage of the fruit.

Benzopyrans

Congenital alveolar rhabdomyosarcoma presenting as a blueberry muffin baby.

Congenital alveolar rhabdomyosarcoma in a newborn was reported. She had multiple subcutaneous tumors and looked like a blueberry muffin baby. From the clinical signs and the histological pictures, a diagnosis of neuroblastoma was considered. Laboratory data showed no abnormalities. Electron microscopic examination revealed myofilament and incipient stria formation. On the basis of these data, a final diagnosis of congenital alveolar rhabdomyosarcoma associated with multiple skin metastases was made.

Diagnosis, Differential

The effect of nitrogen fertilization on forest blueberries.

In a randomized prospective trial with hens it is shown that artificially fertilized forest blueberries are toxic. In a high dose (1.7 1/hens) the birds died within 72 hours (p less than 0.01). In a lower dose (0.33 1/hens) all survived but a lifelong change in the egg production occurred implying defects of the shells (p less than 0.0000001). It seems that fertilizers change the microflora in the ground, which releases soluble aluminiumnitrate which is absorbed in the berries. Aluminium poisoning has been found to have clinical significance in dialysis patients and probably also in senile dementia. In smaller doses it can be difficult to establish this relation as the symptoms can have a latency period of more than 10 years.

Aluminum

[The phenolics of fruits. VIII. Changes in flavonol concentrations during fruit development (authors transl)].

In black currants the contents of quercetin- and especially of myricetin glycosides increased during ripeness very considerably. The myricetin level in ripe fruits of cultivated blueberries and the quercetin level in ripe berries of privet was higher than in unripe. However, the concentrations of kaempferol and quercetin glycosides were in most of the fruits (e.g. red and white currants, sour cherries, plums, cultivated blueberries, elderberries) smaller than in unripe fruits. Anthocyanin formation is therefore not combined with an accumulation of flavonols (perhaps with the exception of myricetin).

Flavonoids

Vitamin C deficiency in growing willow ptarmigan (Lagopus lagopus lagopus).

Willow ptarmigan chicks raised on a diet containing 265 mg ascorbic acid/kg develop scury-like symptoms and die by 4 weeks of age. If blueberry plants are given as an ad libitum supplement to this diet, the malady is prevented. We have described the clinical, pathological and histological changes which accompany this malnutrition and conclude that they are in accord with the description of scurvy in guinea pig and man. Biochemical determination of ascorbic acid synthesis in the kidney of ptarmigan chicks indicated a rate of synthesis five times that found in livers of growing white rats. Blueberry plants and many other plants found in the natural diet of ptarmigan chicks contain 2,000 to 5,000 mg ascorbic acid/kg dry weight. Feeding experiments showed that the pathological signs were avoided and that already afflicted chicks recovered if the vitamin C content of the diet was raised to 750 mg/kg dry weight of food. Since the food intake of the chicks was 5 to 8 g/day the daily requirement of external vitamin C is about 150 mg/kg body weight. To our knowledge this is the first example of an animal which, while producing vitamin C itself, requires substantial amounts of external vitamin C to survive.

Animals

[Congenital cutaneous Langerhans histiocytosis. Apropos of 7 cases].

Seven cases of congenital Langerhans' cell histiocytosis (LH) are reported, with emphasis on clinical and immunohistochemical features. This is a polymorphic disease at birth. In 4/7 cases, the diffuse, generalized rash could be classified as cutaneous Letterer-Siwe disease (LSD); 3/4 remained purely cutaneous and healed in less than 3 months; whereas the fourth-one persisted, pulmonary lesions appeared, and the infant died on his 40th day. In 3/7 cases, the clinical diagnosis at birth was either a Blueberry Muffin Baby (BMB) or Hashimoto-Pritzker type LH (HPLH); the lesions healed rapidly, although one cas was contradictory: typical BMB at birth, histology mimicking a monoblastic cutaneous leukemia, no T.O.R.C.H. syndrome, normal bone marrow, immunophenotyping of LH, auto-involution; 2/3 were MZ twins, both with few lesions. We would like to stress the fact that the clinical spectrum of LH should include BMB, which, however, in most cases must be considered a differential diagnosis. Regarding cutaneous congenital LH, an eponymic classification (LSD, HPLH) is difficult to follow strictly, because overlapping pictures are observed. There is a wide spectrum of cutaneous congenital LH. The main problem at birth is the lack of prognostic criteria. Neither the presence of the rash at birth, nor its type and extension, is necessarily evidence of risk of systemic disease. Cases of HPLH involute, as also do cases of cutaneous LSD, and the "Blueberry Muffin" type of LH; overlapping clinical aspects exist. Histopathological data, electron microscopy or immunohistochemistry, define LH, but they do not enable the outcome to be predicted.(ABSTRACT TRUNCATED AT 250 WORDS)

Diagnosis, Differential

[Laparoscopic diagnosis and evaluation of danazol therapy of endometriosis in sterility].

In twenty two sterile women, endometriosis externa were diagnosed and classified laparoscopically. Classifying the severity of endometriosis by Beecham's and/or Sugimoto's classification, 17 cases were declared mild and only 5 cases severe (one was stage IV and the others were stage III by Beecham's classification, and 4 were grade III by Sugimoto's classification). All these patients were treated with Danazol (Bonzol) 400 mg/day or 300 mg/day for about 24 weeks. Four weeks after the termination of Danazol administration, the effects of Danazol therapy were examined according to the number of blueberry spots, adhesion severity and chocolate cyst size under re-laparoscopy, as well as dysmenorrhea and other complaints. The results were as follows: Dysmenorrhea was relieved in 60.0% of 15 cases, Blueberry spots were decreased or paled in 88.9% of 18 cases, Adhesion was weakened or partially separated spontaneously in 71.4% of 7 cases, Chocolate cyst size became smaller in 85.7% of 7 cases, The patient complained of mild side-effects, namely acne in 45.5% of cases, a weight gain of more than 2 kg in 54.5%, and hoarseness developed in one. The results of laboratory tests performed before and during the treatment remained slightly abnormal or within normal limits.

Adult

[Laparoscopic diagnosis and evaluation of danazol or gestrinone therapy for endometriosis in sterility].

In forty-six sterile women, endometriosis externa was diagnosed and classified laparoscopically. 21 patients were treated with Gestrinone 5mg-10mg/week and 25 patients were treated with Danazol 300mg-400mg/day for 6 months. The effects of these hormonal treatments were evaluated by second-look laparoscopy according to adhesion severity, number of blueberry spots and chocolate cyst size, as well as dysmenorrhea and other complaints. The results were as follows; 1) Dysmenorrhea was relieved in 60.0% of the Gestrinone-treated group and 45.5% of the Danazol group. 2) Adhesion was weakened or partially separated spontaneously in 66.7% of the Gestrinone group and 63.6% of the Danazol group. 3) Blueberry spots decreased in number or paled in 61.9% of the Gestrinone group and 75.0% of the Danazol group. 4) Chocolate cyst size became smaller in 60.0% of the Gestrinone group and 77.8% of the Danazol group. 5) Peritoneal fluid volume was not decreased after the hormonal treatments but the prostaglandin E2 concentration in peritoneal fluid was decreased (p less than 0.05) after Gestrinone therapy. 6) The patients complained of some side effects, liver function especially was disturbed in 48.0% of the Danazol group and 9.5% of the Gestrinone group. Hoarseness was complained of in 33.3% of the Gestrinone group and 12.0% of the Danazol group. 7) Finally, 23.8% of the Gestrinone group and 28.0% of the Danazol group conceived after the hormonal treatments.

Adult