Depth structures the community of arbuscular mycorrhizal fungi amplified from grapevine (Vitis vinifera L.) roots.

The diversity of arbuscular mycorrhizal fungi (AMF) associating with grapevines has been determined previously, yet little is known of how the community in roots is shaped by depth in the soil or where roots occur in different management zones of the vineyard (vine row versus alley). The influence of depth, management zone, and time of year on the community of AMF in grape roots was examined. I also tested the potential influence of the alley vegetation on AMF in grapevines by comparing the taxa amplified from roots of other plants retrieved from the alley surface soil to those from grapevines growing in the same zone. Depth shaped the AMF community in grapevine roots more than the management zone based on dissimilarity among all grapevine samples. Time of the growing season did not, although AMF taxa richness was greater in grapevine roots collected in late summer (veraison) than it was in late spring (bloom). The number of abundant AMF taxa in grapevine roots from the uppermost soil depth in the vine row was substantially lower in late spring than in late summer, and this was related to high soil nitrate in late spring. The alley vegetation comprised primarily grass, and clover plants harbored a different AMF community in roots than did intermingled grapevine roots. The change in the AMF community in a single perennial host (grape) that occurred with depth in this study resulted from a shift among common taxa as opposed to the appearance of unique taxa in the subsoil.

Developmental Dynamics of Meloidogyne hapla in Washington Wine Grapes.

Meloidogyne hapla is the most prevalent plant-parasitic nematode in Washington state wine grape vineyards. Understanding the developmental dynamics of M. hapla can improve the timing of diagnostic sampling and nematicide application. Three Vitis vinifera vineyards in Washington were sampled March 2015 to March 2017 to determine the developmental dynamics of M. hapla by measuring second-stage juveniles (J2) in soil, eggs and adult females in roots, and fine root tips. A model of M. hapla J2 development based on soil growing degree days using a base temperature (Tb) of 0°C (GDDsoil) and a start date of 1 March was developed. This model was validated at two additional vineyards in Washington and was robust with R2 values > 0.74. M. hapla has one generation per year and overwinters primarily as the J2 infective stage. Juvenile populations declined after 1 March, reaching their lowest density in early July and reaching a maximum density over the winter. M. hapla egg and root tip densities reached a maximum in early August. The number of females per root tip did not vary throughout the year. A single generation with defined peaks in J2 population densities will allow for specific timing of nematicide interventions.

Impact of Grapevine (Vitis vinifera) Varieties on Reproduction of the Northern Root-Knot Nematode (Meloidogyne hapla).

One of the most commonly encountered plant-parasitic nematodes in eastern Washington Vitis vinifera vineyards is Meloidogyne hapla; however, limited research exists on the impact of this nematode on V. vinifera. The objectives of this research were to determine the impact of M. hapla on Chardonnay and Cabernet Sauvignon vine establishment and to determine the host status of V. vinifera varieties/clones predominantly grown in Washington to M. hapla. In a microplot experiment, Chardonnay and Cabernet Sauvignon vines were planted into soil inoculated with different densities of M. hapla; population dynamics of M. hapla and vine performance were monitored over 3 yr. In greenhouse experiments, several clones representing five V. vinifera varieties, Chardonnay, Riesling, Cabernet Sauvignon, Merlot, and Syrah, were evaluated as hosts for M. hapla. In both microplot and greenhouse experiments, white varieties were significantly better hosts than red varieties. In the greenhouse experiments, Chardonnay and Riesling had 40% higher reproduction factor values than Syrah and Merlot, however, all varieties/clones screened were good hosts for M. hapla (reproduction factors > 3). In the microplot experiment, M. hapla eggs/g root were 4.5 times greater in Chardonnay compared to Cabernet Sauvignon 3 yr after planting but there was no evident impact of M. hapla on vine establishment.

Spatial Distribution of Plant-Parasitic Nematodes in Semi-Arid Vitis vinifera Vineyards in Washington.

The most commonly encountered plant-parasitic nematodes in eastern Washington Vitis vinifera vineyards are Meloidogyne hapla, Mesocriconema xenoplax, Pratylenchus spp., Xiphinema americanum, and Paratylenchus sp.; however, little is known about their distribution in the soil profile. The vertical and horizontal spatial distribution of plant-parasitic nematodes was determined in two Washington V. vinifera vineyards. Others variables measured in these vineyards included soil moisture content, fine root biomass, and root colonization by arbuscular mycorhizal fungi (AMF). Meloidogyne hapla and M. xenoplax were aggregated under irrigation emitters within the vine row and decreased with soil depth. Conversely, Pratylenchus spp. populations were primarily concentrated in vineyard alleyways and decreased with depth. Paratylenchus sp. and X. americanum were randomly distributed within the vineyards. Soil water content played a dominant role in the distribution of fine roots and plant-parasitic nematodes. Colonization of fine roots by AMF decreased directly under irrigation emitters; in addition, galled roots had lower levels of AMF colonization compared with healthy roots. These findings will help facilitate sampling and management decisions for plant-parasitic nematodes in Washington semi-arid vineyards.

High vitamin D3 diet administered during active colitis negatively affects bone metabolism in an adoptive T cell transfer model.

Decreased bone mineral density (BMD) represents an extraintestinal complication of inflammatory bowel disease (IBD). Vitamin D3 has been considered a viable adjunctive therapy in IBD. However, vitamin D3 plays a pleiotropic role in bone modeling and regulates the bone formation-resorption balance, depending on the physiological environment, and supplementation during active IBD may have unintended consequences. We evaluated the effects of vitamin D3 supplementation during the active phase of disease on colonic inflammation, BMD, and bone metabolism in an adoptive IL-10-/- CD4⁺ T cell transfer model of chronic colitis. High-dose vitamin D3 supplementation for 12 days during established disease had negligible effects on mucosal inflammation. Plasma vitamin D3 metabolites correlated with diet, but not disease, status. Colitis significantly reduced BMD. High-dose vitamin D3 supplementation did not affect cortical bone but led to a further deterioration of trabecular bone morphology. In mice fed a high vitamin D3 diet, colitis more severely impacted bone formation markers (osteocalcin and bone alkaline phosphatase) and increased bone resorption markers, ratio of receptor activator of NF-κB ligand to osteoprotegrin transcript, plasma osteoprotegrin level, and the osteoclast activation marker tartrate-resistant acid phosphatase (ACp5). Bone vitamin D receptor expression was increased in mice with chronic colitis, especially in the high vitamin D3 group. Our data suggest that vitamin D3, at a dose that does not improve inflammation, has no beneficial effects on bone metabolism and density during active colitis or may adversely affect BMD and bone turnover. These observations should be taken into consideration in the planning of further clinical studies with high-dose vitamin D3 supplementation in patients with active IBD.

Agomelatine improves sleep in a patient with fatal familial insomnia.

A young patient with FFI was started on agomelatine 25 mg to medicate nocturnal insomnia. Under this treatment sleep efficiency was improved, slow wave sleep was high and awakenings during sleep period time were far less than before. Clinically the patient was less restless during nighttime.

Comparison of cryopreservation methods on T-cell responses to islet and control antigens from type 1 diabetic patients and controls.

BACKGROUND: Type 1 diabetes (T1D) is a cell-mediated autoimmune disease characterized by destruction of the pancreatic islet cells. The use of cryopreserved cells is preferable to the use of freshly isolated cells to monitor clinical trials to decrease assay and laboratory variability. METHODS: The T-Cell Workshop Committee of the Immunology of Diabetes Society compared two widely accepted T-cell freezing protocols (warm and cold) to freshly isolated peripheral blood mononuclear cells from patients with T1D and controls in terms of recovery, viability, cell subset composition, and performance in functional assays currently in use in T1D-related research. Nine laboratories participated in the study with four different functional assays included. RESULTS: The cold freezing method yielded higher recovery and viability compared with the warm freezing method. Irrespective of freezing protocol, B cells and CD8+ T cells were enriched, monocyte fraction decreased, and islet antigen-reactive responses were lower in frozen versus fresh cells. However, these results need to take in to account that the overall response to islet autoantigens was low in some assays. CONCLUSIONS: In the current study, none of the tested T-cell functional assays performed well using frozen samples. More research is required to identify a freezing method and a T-cell functional assay that will produce responses in patients with T1D comparable to responses using fresh peripheral blood mononuclear cells.

Nitrate Uptake and Transport Properties of Two Grapevine Rootstocks With Varying Vigor.

In viticulture, rootstocks are essential to cope with edaphic constraints. They can also be used to modulate scion growth and development to help improve berry yield and quality. The rootstock contribution to scion growth is not fully understood. Since nitrogen (N) is a significant driver of grapevine growth, rootstock properties associated with N uptake and transport may play a key role in the growth potential of grafted grapevines. We evaluated N uptake and transport in a potted system using two grapevines rootstocks [Riparia Gloire (RG) and 1103 Paulsen (1103P)] grafted to Pinot noir (Pommard clone) scion. Combining results of nitrate induction and steady-state experiments at two N availability levels, we observed different responses in the uptake and utilization of N between the two rootstocks. The low vigor rootstock (RG) exhibited greater nitrate uptake capacity and nitrate assimilation in roots after nitrate resupply than the more vigorous 1103P rootstock. This behavior may be attributed to a greater root carbohydrate status observed in RG for both experiments. However, 1103P demonstrated a higher N translocation rate to shoots regardless of N availability. These distinct rootstock behaviors resulted in significant differences in biomass allocation between roots and shoots under N-limited conditions, although the overall vine biomass was not different. Under sufficient N supply, differences between rootstocks decreased but 1103P stored more N in roots, which may benefit growth in subsequent growing seasons. Overall, greater transpiration of vines grafted to 1103P rootstock causing higher N translocation to shoots could partially explain its known growth-promoting effect to scions under low and high N availability, whereas the low vigor typically conferred to scions by RG may result from the combination of lower N translocation to shoots and a greater allocation of biomass toward roots when N is low.

Novel N-glycosylation in eukaryotes: laminin contains the linkage unit beta-glucosylasparagine.

The linkage unit to protein of N-linked carbohydrate in eukaryotic glycoproteins consists of N-acetylglucosamine, coupled to the amido nitrogen of asparagine. Additional N-glycosyl linkage units have been unequivocally proven to exist only in the cell surface glycoproteins of various bacteria. Based on immunological analyses, isolation and chemical characterization, we report that one of these units, namely glucose linked to asparagine, exists in the mammalian protein laminin, an extracellular basement membrane component. This finding and the occurrence of identical disaccharide structures in archaebacterial cell surface glycoproteins and mammalian basement membrane protein complexes points towards a conserved and distinct function of these extracellular structural elements. In addition, a method is described to uncover a masked epitope in fixed tissues by chemical O-deglycosylation. This has allowed to morphologically localize the antigen beta-Glc-Asn by immunofluorescence to the basement membranes of kidney glomeruli.

The diversity of arbuscular mycorrhizal fungi amplified from grapevine roots (Vitis vinifera L.) in Oregon vineyards is seasonally stable and influenced by soil and vine age.

The diversity of arbuscular mycorrhizal fungi (AMF) in 10 Oregon vineyards was assessed by examining spores in soil and amplifying mycorrhizal DNA from roots. Seventeen spore morphotypes were found in soil, including seven species in the Acaulosporaceae. Eighteen phylotypes were amplified from grape roots with AM1 and NS31 primers, and clones were dominated by Glomus spp. (> 99%). A few clones (< 1%) representing a single phylotype within Gigasporaceae, and a single clone within Archaeosporaceae were amplified from roots with AM1-NS31 primers. A separate experiment employing known proportions of grape roots colonized by Glomus intraradices or by Gigaspora rosea showed that fungi within Gigasporaceae might be underrepresented in clone abundance when Glomus spp. co-occur in roots. No clones representing fungi within the Acaulosporaceae were amplified from vineyards, although specific fungi within Acaulosporaceae were shown to colonize Pinot noir roots in sterilized soil and were amplified from these roots. Four Glomus phylotypes, including G. intraradices, were found in roots from all 10 vineyards, and these fungi accounted for 81% of clones. AMF phylotypes amplified from roots did not change during the growing season, although six phylotypes varied with soil type. The presence of three phylotypes was affected by vineyard age, and phylotype richness appeared to decline as vineyard age increased beyond 20 y. PCA analysis supported the hypothesis that the AMF community is different in red-hill soils than in valley soils and indicated certain phylotypes might be associated with lower soil and vine nutrient status. However, the changes in the AMF community in grape roots across vineyards were subtle because most root samples were dominated by the same three or four phylotypes. A separate analysis using primers to amplify AMF from the Archeasporaceae/Paraglomeraceae showed most root samples also were colonized by at least one Paraglomus or Archaeospora phylotype.

Absolute testing of the reference surface of a Fizeau interferometer through even/odd decompositions.

Absolute testing of spherical surfaces is a technological necessity because of increased accuracy requirements. In a Fizeau setup, the main part of the interferometer deviations thereby comes from the reference surface. We demonstrate the validity of an absolute testing procedure for the reference surface that has been proposed earlier. The procedure relies on the decomposition of the surface deviations into odd and even parts and could be used in partially coherent illumination. The odd deviations are obtained from a basic and a 180 degree-rotated position of an auxiliary sphere, and the even deviations can be measured with the help of a cat's eye position in double pass using an opaque half screen in the interferometer aperture.

Epidemiology of Xiphinema americanum and Tomato ringspot virus on Red Raspberry, Rubus idaeus.

Population dynamics of Xiphinema americanum and transmission of Tomato ringspot virus (ToRSV) were studied in a red raspberry field in Washington State. Population densities of X. americanum were highest in the winter, lowest in the summer, and were correlated with precipitation (R2 = 0.42). All nematode stages were present throughout the year. Gravid females were observed only in the spring, indicating one generation per year. The sequence of the coat protein of the ToRSV from this field was similar to those of other raspberry isolates from the Pacific Northwest. Cucumber seedlings were planted in soil collected monthly from the field and were evaluated for nematode transmission of ToRSV by enzyme-linked immunosorbent assay (ELISA). The proportion of assay plants infected with ToRSV was negatively correlated with nematode densities (R2 = 0.31). In another study, ToRSV was detected by ELISA in fine roots of raspberry plants 5 months after planting in field soil infested with viruliferous nematodes, in all subterranean portions of plants after 12 months, and in all aerial portions the second year. The rate of spread of ToRSV in a raspberry field was 70 cm per year. These results suggest that the rate of ToRSV spread is limited by systemic spread of virus in plants when nematode-infested soil is not transported in the field.

Gamma-COP, a coat subunit of non-clathrin-coated vesicles with homology to Sec21p.

Constitutive secretory transport in eukaryotes is likely to be mediated by non-clathrin-coated vesicles, which have been isolated and characterized [(1989) Cell 58, 329-336; (1991) Nature 349, 215-220]. They contain a set of coat proteins (COPs) which are also likely to exist in a preformed cytosolic complex named coatomer [(1991) Nature 349, 248-250]. From peptide sequence and cDNA structure comparisons evidence is presented that one of the subunits of coatomer, gamma-COP, is a true constituent of non-clathrin-coated vesicles, and that gamma-COP is related to sec 21, a secretory mutant of the yeast Saccharomyces cervisiae.

Interlaboratory bilirubin variability.

Stabilized liquid quality control sera, some with elevated bilirubin concentrations, were distributed to laboratories for total and conjugated (direct) bilirubin analyses. Interlaboratory variability was high; for example, results ranged from 10.9 to 24.0 mg/100 ml for a serum with a mean bilirubin concentration of 18.1 mg/100 ml. Coefficients of variation were typically 10% to 12% for total bilirubin ad approximately 24% for conjugated bilirubin analyses. It is believed that these stabilized sera, when used as quality control materials, can help reduce the unacceptably large interlaboratory variability found in this study.

Neonatal pneumoperitoneum without significant adventitious pulmonary air: use of metrizamide to rule out perforation of the bowel.

Pneumoperitoneum in the neonate may be due to air that has dissected from the chest. Four infants, in whom pneumoperitoneum, from thoracic air dissection, occurred in the absence of pneumothorax or pneumomediastinum are reported. The absence of abnormal collections of air in the chest is of no value in deciding whether a pneumoperitoneum is due to bowel perforation or air dissecting from the chest. Contrast bowel studies are needed in all cases of pneumoperitoneum in the infant to distinguish bowel perforation from intrathoracic origin of the air.

The use of TORCH titers.

Of 603 patients undergoing serologic tests for Toxoplasma, syphilis, rubella, cytomegalovirus, and herpes simplex (TORCH tests) during a 4-year period, 381 (63%) were infants younger than 6 months and 110 (18%) were either the mothers of those infants or pregnant women. The remaining 112 (19%) were older infants and children, male adults, or nonpregnant female adults. Of 381 infants less than 6 months of age who were tested, 323 (85%) had only a single TORCH test. Of 35 (9%) infants who had follow-up titers, 16 (46%) had a specific titer requested whereas in 19 (54%) the entire TORCH battery was repeated. Follow-up titers were infrequently (25%) requested when initial titers were elevated. Infection with a TORCH agent was not confirmed serologically in any of the 603 patients. The pattern of TORCH test use has a poor diagnostic return.

Maternal oral herpes: isolation policy.

Current official recommendations are that parents and personnel with oral herpes lesions be isolated from newborn infants. A survey of 110 neonatal centers, 50% of them replying, showed that most centers do not isolate mothers with oral herpes from their newborn infants. A review of the literature suggest that exposure of infants to personnel or family with oral herpes lesions might occasionally result in disseminated neonatal herpes simplex virus disease. Since the separation of a mother from her newborn infant up to 1 week of ages is a decision that should not be taken lightly, more information is urgently needed to decide whether or not to isolate mothers and hospital personnel with oral herpes lesions from newborn infants. Recommendations to obtain more information concerning these infants are provided.

Fetal and neonatal ventricular arrhythmia.

Ventricular arrhythmia in the perinatal period is observed with greater frequency than reported in the literature. Four cases from the authors' experience and an analysis of the literature are presented. Of the total of 45 cases, nine were detected in utero, three persisted beyond the neonatal period, and two resulted in death with associated disorders. Twenty-four percent of the entire group had serious medical disorders associated with arrhythmia; however, 43% of the group with ventricular tachycardia had major associated disease.

Countercurrent immunoelectrophoresis in group B streptococcal disease.

Countercurrent immunoelectrophoresis was used for the detection of group- and type-specific antigens in the body fluids of 61 infants from St. Louis and Indiana with group B streptococcal infections. Urine concentrated using an Amicon filter yielded the highest percentage of positive results; 81% were positive in the St Louis group. When three body fluids (urine, CSF, and blood) were available, at least one was positive for group B streptococcus in 95% of the cases. This study demonstrates the applicability of this test in a tertiary care facility (St Louis) and in smaller hospitals (Indiana) with access to central laboratory.

Improved survival of ventilated neonates with modern intensive care.

The records of 909 infants who required mechanical ventilation over a 42-month period were reviewed; 68.3% of the infants survived. The highest survival rate was in infants with persistent fetal circulation and hyaline membrane disease, and the lowest in infants with congenital anomalies and miscellaneous diseases. The survival rate for each year of the 3 1/2-year study increased from the previous year. There was also a dramatic increase in the survival rate in infants with a birth weight greater than 1,250 gm.