Outcomes with durvalumab by tumour PD-L1 expression in unresectable, stage III non-small-cell lung cancer in the PACIFIC trial.

BACKGROUND: In the PACIFIC trial, durvalumab significantly improved progression-free and overall survival (PFS/OS) versus placebo, with manageable safety, in unresectable, stage III non-small-cell lung cancer (NSCLC) patients without progression after chemoradiotherapy (CRT). We report exploratory analyses of outcomes by tumour cell (TC) programmed death-ligand 1 (PD-L1) expression. PATIENTS AND METHODS: Patients were randomly assigned (2:1) to intravenous durvalumab 10 mg/kg every 2 weeks or placebo ≤12 months, stratified by age, sex, and smoking history, but not PD-L1 status. Where available, pre-CRT samples were tested for PD-L1 expression (immunohistochemistry) and scored at pre-specified (25%) and post hoc (1%) TC cut-offs. Treatment-effect hazard ratios (HRs) were estimated from unstratified Cox proportional hazards models (Kaplan-Meier-estimated medians). RESULTS: In total, 713 patients were randomly assigned, 709 of whom received at least 1 dose of study treatment durvalumab (n = 473) or placebo (n = 236). Some 451 (63%) were PD-L1-assessable: 35%, 65%, 67%, 33%, and 32% had TC ≥25%, <25%, ≥1%, <1%, and 1%-24%, respectively. As of 31 January 2019, median follow-up was 33.3 months. Durvalumab improved PFS versus placebo (primary-analysis data cut-off, 13 February 2017) across all subgroups [HR, 95% confidence interval (CI); medians]: TC ≥25% (0.41, 0.26-0.65; 17.8 versus 3.7 months), <25% (0.59, 0.43-0.82; 16.9 versus 6.9 months), ≥1% (0.46, 0.33-0.64; 17.8 versus 5.6 months), <1% (0.73, 0.48-1.11; 10.7 versus 5.6 months), 1%-24% [0.49, 0.30-0.80; not reached (NR) versus 9.0 months], and unknown (0.59, 0.42-0.83; 14.0 versus 6.4 months). Durvalumab improved OS across most subgroups (31 January 2019 data cut-off; HR, 95% CI; medians): TC ≥ 25% (0.50, 0.30-0.83; NR versus 21.1 months), <25% (0.89, 0.63-1.25; 39.7 versus 37.4 months), ≥1% (0.59, 0.41-0.83; NR versus 29.6 months), 1%-24% (0.67, 0.41-1.10; 43.3 versus 30.5 months), and unknown (0.60, 0.43-0.84; 44.2 versus 23.5 months), but not <1% (1.14, 0.71-1.84; 33.1 versus 45.6 months). Safety was similar across subgroups. CONCLUSIONS: PFS benefit with durvalumab was observed across all subgroups, and OS benefit across all but TC <1%, for which limitations and wide HR CI preclude robust conclusions.

Richieri-Costa-Pereira syndrome: Expanding its phenotypic and genotypic spectrum.

Richieri-Costa-Pereira syndrome is a rare autosomal recessive acrofacial dysostosis that has been mainly described in Brazilian individuals. The cardinal features include Robin sequence, cleft mandible, laryngeal anomalies and limb defects. A biallelic expansion of a complex repeated motif in the 5' untranslated region of EIF4A3 has been shown to cause this syndrome, commonly with 15 or 16 repeats. The only patient with mild clinical findings harbored a 14-repeat expansion in 1 allele and a point mutation in the other allele. This proband is described here in more details, as well as is his affected sister, and 5 new individuals with Richieri-Costa-Pereira syndrome, including a patient from England, of African ancestry. This study has expanded the phenotype in this syndrome by the observation of microcephaly, better characterization of skeletal abnormalities, less severe phenotype with only mild facial dysmorphisms and limb anomalies, as well as the absence of cleft mandible, which is a hallmark of the syndrome. Although the most frequent mutation in this study was the recurrent 16-repeat expansion in EIF4A3, there was an overrepresentation of the 14-repeat expansion, with mild phenotypic expression, thus suggesting that the number of these motifs could play a role in phenotypic delineation.

Glomerella Leaf Spot Caused by a Nonhomothallic Strain of Glomerella cingulata on Highbush Blueberry Nursery Plants in Buenos Aires, Argentina.

In February 2009, irregular-shaped leaf spots affected blueberry (Vaccinium corymbosum L. 'Blue Crisp', 'Misty', and 'Sharp Blue') nursery plants in Buenos Aires. Single-spore cultures on potato dextrose agar and oat agar showed aerial white mycelium that turned light and dark gray, dark brown acervuli with setae, and a salmon-to-orange conidial mass. Septate, dark brown, 62 to 78 µm long setae were abundant in the acervulus. Conidia were unicellular, hyaline, straight, cylindrical, round at the ends, and averaged 15.2 (12.1 to 16.9) × 5.4 (4.9 to 6.2) µm. Dark brown, ovate to clavate, 10.25 × 6.25 µm (9 to 12 × 5 to 8) appressoria with a noticeable pore formed on slides near the edge of the cover glass. Dark subglobose structures were recorded immersed in the culture medium. No asci or ascospores were observed, indicating a nonhomothallic condition. The fungus was identified as Colletotrichum gloeosporioides (Penz.) Penz & Sacc. (teleomorph Glomerella cingulata (Stoneman) Spauld. & H. Schrenk) with traits similar to those already described (1). DNA was obtained from mycelium with a standard DNA extraction kit and the ribosomal, internal transcribed spacer (ITS) 1 and ITS2 regions were PCR amplified and sequenced with primers ITS1 and ITS4 (2). A BLASTN algorithm search revealed 100% identity of the sequence (535 bp long) with G. cingulata/C. gloeosporioides from citrus and mango and one from coffee identified as C. kahawae (GenBank Accession No. JF908919). The nucleotide sequence was deposited in GenBank (Accession No. JQ340087). Pathogenicity was verified on young plants and detached leaves of highbush blueberry 'Emerald', 'Misty', 'O'Neal', and 'Santa Fe', olive (Olea europaea 'Arbequina'), and marketed fruits of apple, mango, orange, and tomato. Disinfected healthy leaves were inoculated with a 9-mm2 mycelial block and incubated at 24°C with 12 h of light. Young plants were infected by placing the disinfected end of the branches within a micropipette tip filled with mycelium and kept under greenhouse conditions. Asymptomatic fruits of apple, mango, orange, and tomato were inoculated by placing a mycelial block on a small wound made on their surface. Detached leaves of highbush blueberry 'Emerald', 'O'Neal', 'Misty', and 'Santa Fe' showed 0.1 to 1.5 × 0.8 to 2 cm necrotic lesions after 3 days, covering 43 to 100% of the 'Emerald' leaf area after 8 days. Young plants of blueberry 'Emerald' and 'Misty' showed 1.5 to 3 cm necrotic lesions, acervuli, a salmon-orange conidial mass, and death of leaves at 25 days. On olive 'Arbequina', leaf necrotic lesions reached 0.1 to 3.5 cm after 5 days. Symptoms developed slowly on infected tomato fruits while inoculated fruits of apple, mango, and orange showed dark brown lesions that measured 2 to 7 × 1 to 3.5 cm at 5 days. No symptoms were observed on controls. The fungus was reisolated from inoculated plant parts. The disease was previously cited in Argentina (3), but to our knowledge, this is the first report of a nonhomothallic strain of G. cingulata from highbush blueberry colonizing and deteriorating fruits of apple, mango, orange, and tomato. References: (1) J. M. E. Mourde. No 315. CMI Descriptions of Pathogenic Fungi and Bacteria. Kew, Surrey, UK, 1971. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990. (3) E.R. Wright et al. OEPP/EPPO Bull. 28:219, 1998.

First Report of Root Rot Caused by Fusarium proliferatum on Blueberry in Argentina.

In 2009, a highbush blueberry (Vaccinium corymbosum L. 'O'Neal') field located in Rojas, Buenos Aires Province showed 30% of plants with dry or dead branches. Disinfected root pieces were placed on water agar and incubated at 24°C. A fungal colony was obtained and purified by successive transfers of an individual hyphal tip from a sparsely growing colony. Colony color and growth rate were evaluated in potato dextrose agar where the fungus produced white-to-pale pink colonies and grew 3.5 cm after 5 days. The fungus was studied on Spezieller Nährstoffarmer agar (2), carnation leaf-piece agar, and KCl agar where it produced abundant single-celled hyaline microconidia in moderate-length chains and in false heads originated from monophialides and polyphialides. Microconidia measured 6 to 12 × 2 to 3 µm (average 8 × 2.3 µm). On KCl, chains of microconidia and tan-to-light cream sporodochia with 3- to 5-septate, slender, relatively straight macroconidia were easily observed after 4 and 10 days, respectively. Macroconidia measured 38 to 48 × 3.5 to 4 µm (average 43.9 × 3.9 µm). Chlamydospores and sclerotia were not present. Data coincided with the description for Fusarium proliferatum (Matsush.) Niremberg ex Gerlach & Niremberg. The isolate was deposited in the IMYZA Microbial Collection as INTA-IMC 144. The fungus was cultured in 100 ml of Czapek-Dox supplemented with sucrose, peptone, yeast extract, sodium nitrate, and vitamins for 4 days. Genomic DNA was obtained with a DNA extraction kit, PCR amplified with primers ITS1 and ITS4 for the internal transcribed spacer (ITS) region of ribosomal genes, and sequenced. The nucleotide sequence (Accession No JF913468) was compared with GenBank records. The sequence shared 99% identity with Accession No HQ113948 for F. proliferatum. Pathogenicity was confirmed in 1-year-old 'O'Neal' plants. A 10-ml suspension (2.4 × 106 conidia/ml in sterile distilled water) was applied to six potted plants grown in sterilized potting mix. Roots were superficially wounded with a needle. Control plants were treated with sterile distilled water. Plants were incubated at 24°C and a 12-h photoperiod. After 90 days, plants showed root rot, leaf chlorosis, and branch necrosis followed by plant death. Control plants remained healthy. F. proliferatum was reisolated from diseased roots of inoculated plants. This fungus was previously cited in Argentina on asparagus (1), corn (1,3), and oat (4). To our knowledge, this is the first report of F. proliferatum as a root pathogen of highbush blueberry in Argentina. References: (1) G. Lori et al. Plant Dis. 82:1405, 1998. (2) H. I. Nirenberg. Releases Fed. Biol. Res. Ctr. Agric. For. (Berlin-Dahlem) 169:1, 1976. (3) D. A. Sampietro et al. Fung. Biol. 114:74, 2010. (4) S. A. Stenglein et al. Plant Dis. 94:783, 2010.

First Report of Fusarium solani Causing Root Rot of Olive in Southeastern Argentina.

In Argentina, olive (Olea europaea L.) is cultivated in the mountainous, warm, arid northwest (Andes range), where Fusarium solani (blue sporodochia) is frequently found to be causing death of nursery and young field plants (1). Recently, olive orchards were established more than 1,600 km to the southeast (Pampas) in a plain with a temperate and humid climate and in the arid Patagonia, both influenced by the Atlantic Ocean. This area includes part of Buenos Aires and Rio Negro provinces. In March 2008, 10-year-old 'Barnea' olive trees with high incidence of root rot, dried leaves, dead branches, and dead plants were observed in the Coronel Dorrego District of Buenos Aires Province, where oat, barley or other cereals are planted between rows of olive trees. Planting material originated from olive nurseries located in Mendoza Province, 1,200 km from Coronel Dorrego. Diseased roots were disinfected in 2% NaOCl and 70% ethanol, cut into small pieces, plated onto rose bengal-glycerin-urea medium, and incubated at 20°C with a 12-h photopheriod. A fungus was purified through successive transfers of hyphal tips from the margin of a sparsely growing colony onto 2% water agar (2). Colonies grown on Spezieller Nährstoffarmer agar (3) and carnation leaf-piece agar were used for morphological identification, and those on grown on potato dextrose agar were used for evaluation of pigmentation and colony growth rate. Sporodochium color, cream, was typical of F. solani (Mart.) Sacc. This isolate was deposited in the IMYZA Microbial Collection as INTA-IMC 73. Mycelium was cultured in liquid Czapek-Dox medium supplemented with sucrose, peptone, yeast extract, sodium nitrate, and vitamins for 4 days and fungal DNA was obtained with a DNA extraction kit. Primers ITS1 and ITS4 were used to amplify the internal transcribed spacer (ITS) region of ribosomal genes. The purified PCR product was sequenced and the DNA sequence compared with GenBank records. The sequence shared 100% identity with 27 entries for F. solani and 97% identity with F. solani obtained from olive in Nepal (4), corresponding to EU912432 and EU912433. The nucleotide sequence was registered in GenBank as JF299258. Pathogenicity was confirmed on 'Manzanilla' plants at the eight-leaf stage. Pieces of water agar with mycelium were applied to small wounds at the stem base and on roots of 10 plants and were covered with cotton soaked in sterile distilled water. Plants were incubated at 20°C and a 14-h photoperiod. On control plants, water agar pieces without mycelium were applied to the wounds. After 33 days, inoculated plants showed dark brown lesions (average length 1.4 cm) and leaf chlorosis. Two plants showed wilting with leaves remaining attached to branches. F. solani was reisolated from roots and stem bases of inoculated plants. Controls remained asymptomatic. To our knowledge, this is the first report of F. solani occurring on olive in the temperate part of the Pampas of Argentina where cereals, which are susceptible to Fusarium species, are grown with olive trees. Sporodochium color (cream) of these isolates differed from the blue color of previously reported isolates of F. solani on olive in northwestern Argentina (1). References: (1) S. Babbitt et al. Plant Dis. 86:326, 2002. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (3) H. I. Nirenberg. Releases Fed. Biol. Res. Center Agric. For. (Berlin-Dahlem) 169:1, 1976. (4) A. M. Vettraino et al. Plant Dis. 23:200, 2009.

First Report of Sclerotinia Rot on Blueberry Caused by Sclerotinia sclerotiorum in Argentina.

In October 2007, blighted shoots were observed on highbush blueberry (Vaccinium corymbosum L. cv. O'Neal) plants in La Plata, Buenos Aires Province, Argentina. Isolations from surface-disinfested shoots onto carrot agar and Spezieller Nahrstoffarmer Agar (SNA) consistently yielded white colonies that produced black sclerotia, mainly near the edge of the culture plates, after 7 days. Sclerotia were transferred to SNA tubes and kept at 5°C for several months. The germination of sclerotia produced numerous 6 mm long initials, stipitate pale brown cup-shaped apothecia (10 × 6 mm) with eight-spored asci (137 × 7 µm) at 18°C and continuous light conditions. Asci with uniseriate ascospores were cylindrical and narrow at the base. Ascospores (11 to 12 × 4 µm) were hyaline, unicellular, smooth, and ellipsoid. This isolated fungus was morphologically identified as Sclerotinia sclerotiorum (Lib.) de Bary (2,3). The isolate was deposited in the IMYZA Microbial Collection as INTA-IMC 87. Mycelium was cultured in 100 ml of Czapek's-Dox medium, supplemented with sucrose, peptone, yeast extract, sodium nitrate, and vitamins (1), for 3 days and fungal DNA was obtained using a DNA extraction kit. ITS1 and ITS2 of ribosomal genes were amplified by PCR using universal primers (4) and the PCR product was sequenced. A BLAST algorithm search revealed 100% identity of the sequence with 12 GenBank entries for S. sclerotiorum. The nucleotide sequence was deposited in the GenBank with Accession No. JF277567. Pathogenicity testing was achieved by placing detached leaves of cvs. Emerald, Misty, and Start on water agar (WA) plates, inoculating with 9-mm2 mycelial blocks, and incubating at 20°C with 12 h of light. Young shoots of highbush blueberry, Misty and O'Neal, were inoculated by the cut shoot method with micropipette tips filled with mycelium and kept under greenhouse conditions at 24°C and 14 h of light. On control plants, WA blocks or WA-filled micropipette tips were used. Leaf blight was observed after 5 to 6 days and sclerotia appeared after 7 days on inoculated tissues. Shoot blight was recorded on inoculated plants after 5 days. The fungus was reisolated from inoculated tissues, with no symptoms showing on controls. To our knowledge, this is the first report of Sclerotinia rot caused by S. sclerotiorum in blueberry in Argentina. References: (1) J.F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Hoboken, NJ, 2006. (2). J. E. M. Mourde and P. Holliday. No. 513 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Kew, Surrey, UK, 1976. (3) S. Umemoto et al. Gen. Plant Pathol. 73:290, 2007. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Hop Downy Mildew Caused by Pseudoperonospora humuli in Argentina.

In Argentina, hop downy mildew disease caused by Pseudoperonospora humuli first appeared in Alto Valle of Rio Negro and Neuquen (1957), Mar del Plata (1962), and El Bolson (1963) (1). The disease occurred in the hop (Humulus lupulus L.) planting areas of El Bolson (Rio Negro) and Lago Pueblo (Chubut) in 2002 and 2003. Surveys were conducted in 30 commercial hop fields from December 2002 to March 2003 to estimate disease incidence and susceptibility of cultivars planted in these fields. Hop fields were divided into five sections and 100 plants were randomly selected and assessed for the presence of disease. Symptoms that were observed in early spring included dark brown rootstocks and primary basal spikes (stunted plants with pale and curled leaves), which are characteristic of systemic infection. Later in the season, secondary infections were characterized by dark purple-to-black lesions on leaves, flowers, cones, and lateral and terminal spikes. Plant symptoms and fungal morphological markers (dichotomously branched sporangiophores; ellipsoid and papillate sporangia) agreed with hop downy mildew disease and the fungus P. humuli. Yield loss was estimated as the reduction in yield compared with the 2001-2002 season observed from five hop growers. On December 10, 70% of the hop fields had greater than 50% disease incidence and seven fields reached 100% incidence. The reduction in cone yield varied between 20 and 34% in fields without a rootstock fungicide treatment. One field with a rootstock fungicide treatment (mefenoxam, copper oxiclorure, phosphorous acid, copper sulfate, and fosetyl-Al) and regular fungicide applications had a 30% increase in cone yield compared with 2001-2002. Systemically infected plants were recorded for hop cvs. Bullion, Cascade, CEZ, GS-19, Hallertauer Mfr., Nugget, Spalt, and Traful. Previously, Cascade was rated as a resistant cultivar to the root systemic infection (1). To our knowledge, this is the first record of a hop downy mildew outbreak in Argentina during the last 30 years. Reference: (1) L. Leskovar. El Lúpulo: Su Cultivo y Procesamiento. Hemisferio Sur. Buenos Aires, 1978.

Occurrence of Late Leaf Rust Caused by Pucciniastrum americanum in Red Raspberry (Rubus idaeus) in Buenos Aires, Córdoba, and Entre Ríos, Argentina.

In the fall of 2003, severity of late leaf rust in leaves and fruits of red raspberry (Rubus idaeus L.) reached 50% in Buenos Aires (Azul, Baradero, Capilla del Señor, Gral. Rodríguez, Mar del Plata, and Tandil), Córdoba (Villa de Las Rosas), and Entre Ríos (Concordia). The south of Argentina, Río Negro (El Bolsón), and Chubut (El Hoyo, Lago Puelo) remained rust free. The abaxial side of the field infected leaves had pustules filled with masses of yellow spores. Chlorotic areas corresponded in the adaxial side. Urediospores were vacuum harvested from field infected leaves collected in the Tandil area and placed onto a healthy 1-year-old greenhouse-grown plant (cv. Heritage). Spores from a single pustule were increased on plants of the same cultivar. Spores were studied with optic and electronic microscopy. Uredial ostiolar cells were warted, laterally free, and constricted in the middle. The obovoid, echinulate urediospores, from infected leaves averaged 24 × 16 µm (16 to 30 × 11 to 21 µm). Morphological characteristics and spore measurements agreed with those reported for Pucciniastrum americanum (1). Urediniospores were suspended in mineral oil and sprayed onto three raspberry cultivars that were maintained in a darkened mist chamber at 20°C for 48 h and the transferred to a 20°C and 12-h light cycle chamber. Control plants were inoculated with sterile water. There were three replicate plants of each treatment. After 11 days, large sporulating uredia (0.5 mm) were produced on inoculated leaves of cv. Autumn Bliss and smaller uredia (0.1 to 0.3 mm) were produced on cv. Heritage. There were necrotic flecks and the least and smallest uredia were produced on cv. Himbo Queen. No symptoms were present in control plants. To our knowledge, this is the first report of P. americanum causing disease on raspberry in Argentina. Reference: (1) G. F. Laundon and A. F. Rainbow. Pucciniastrum americanum. No. 210 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, England, 1969.

Occurrence of Fusarium solani on Blueberry in Argentina.

Blueberry plants with root rot and sudden death symptoms were collected in Concordia, Entre Ríos. Diseased roots were disinfected by immersion in 0.5% NaOCl for 2 min, cut into pieces, transferred to carrot agar (CA), and maintained at 20 ± 2°C and 12 h of near UV light (Philips Black Light lamps TL 40W/08). Conidia were identified with an Olympus BX-51 optical microscope by using a CoolSNAP Pro digital kit with image-pro plus and color digital camera (Media Cybernetics, Inc., Silver Spring, MD). Macroconidia length was variable, 3.9 µm and as much as 29 µm (1 to 5 septa), and microconidia measured 3.8 × 11 µm. Fungal description agrees with Fusarium solani (1). Pathogenicity of the purified isolate was evaluated on 2-month-old plants (cvs. Misty and Sharp Blue). The purified, grayish white isolate was grown on CA for 7 days, and mycelial plugs were placed next to the base of wounded stem and roots immediately below the potting mix soil line. Plants were maintained in the dark at 20 ± 2°C and 90% humidity for 48 h, and then transferred to 12 h of light. Wounded plants with CA plugs served as controls. Dark spots along the stem and root and stem rot appeared 7 to 21 days after inoculation. Controls remained symptomless. The fungus was reisolated from inoculated plants. Fusarium sp. was previously cited (2). To our knowledge, this is the first report of F. solani on blueberry in Argentina. References: (1) C. Booth. Fusarium. Laboratory Guide to the Identification of the Major Species. CMI, Kew, England, 1977. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.

Occurrence of Blight Caused by Sclerotium rolfsii on Lolium perenne in Argentina.

A perennial ryegrass (Lolium perenne L.) lawn located at Castelar (Buenos Aires Province) showed disease symptoms during the summer of 2003. Chlorotic patches as much as 15 cm in diameter appeared on the lawn. Later, dead plants with white mycelia developing on the crown and surrounding soil occurred at the periphery of the rings. Plants showed leaf chlorosis and crown and root rot. No sclerotia developed on plant organs. Diseased plants were collected, washed with running tap water for 4 h, and disinfested in 5% NaOCl for 2 min. Pieces, 3 to 5 mm long from symptomatic leaves, crowns, and roots, were incubated on 2% potato dextrose agar (PDA) at 22 to 25°C with a 12-h light/dark cycle. Mycelia growing on the soil surface was transferred to PDA and incubated under the same conditions. After 3 to 4 days, white, conspicuous mycelia that produced sclerotia grew from diseased tissue pieces and soil mycelial samples. Sclerotia were nearly spherical, 1 to 2 mm in diameter, white but turning brown with age, and produced in large numbers over the entire colony surface. Primary hyphae showed clamp connections at the septa. A pathogenicity test was performed with 20 1-month-old plants of L. perenne grown in a 1:1 (v/v) mixture of sand and soil contained in 24- × 17- × 4-cm plastic trays. Seven-day-old fungal cultures grown on PDA were cut into 1- cm2 pieces and placed among the plants on the substrate. Each tray was inoculated with seven inoculum pieces. Five trays of plants were inoculated with the fungus, and plants in five trays that served as controls had only sterile pieces of PDA placed on the substrate. All plants were maintained at 25°C and watered frequently. First symptoms, consisting of chlorosis, were observed 4 days after inoculation. Of the plants, 34, 59, 60, 65, and 70% developed symptoms 6, 9, 14, 17, and 21 days after inoculation, respectively. Control plants remained healthy. The fungus was reisolated from diseased plants and identified as Sclerotium rolfsii Sacc. (teleomorph Athelia rolfsii (Curzi) C.C. Tu & Kimbr.) on the basis of morphological and cultural characteristics (3,4). The disease has been observed causing stalk rot on perennial ryegrass in the United States (1) and Australia (2). To our knowledge, this is the first report of S. rolfsii causing disease on L. perenne in Argentina. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society. St. Paul, MN. 1989. (2) D. F. Farr et al. Fungal Databases. Systematic Botany and Mycology Laboratory. Online publication. ARS, USDA, 2007. (3) J. E. M. Mordue. No. 410 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, UK, 1974. (4) Z. K. Punja and A. Damiani. Mycologia 88:694, 1996.

First Report of Blueberry Botrytis Blight in Buenos Aires, Entre Ríos, and Córdoba, Argentina.

Since 2003, a new field disease has been observed on several cultivars of highbush blueberry (Vaccinium corymbosum L.) in Buenos Aires (Baradero, Colonia Urquiza, Lima, Mercedes, and San Pedro), Entre Ríos (Concordia, Gualeguaychú, and Larroque), and Córdoba (Capilla del Monte and La Cumbre). Infected flowers turned brown to tan with a water-soaked appearance and shriveled up. Blighted flowers typically did not produce fruits; even an entire cluster of berries could be aborted. A chlorotic area, that later became necrotic and turned light brown, developed when leaves were in contact with blighted flowers. A watery rot developed on fruit occasionally before harvest but more generally after harvest. Infected tender green twigs also became blighted, with leaf tissue becoming brown to black. Older twigs and stems were also blighted. Abundant, gray mycelium with conidial masses developed on all affected tissues under moist conditions. Sections of infected leaves, twigs, stems, flowers, and fruits were surfaced sterilized with 0.2% NaOCl, plated on 2% potato dextrose agar (pH 7), and incubated at 22°C. Pure cultures formed a whitish dense mycelial mat and turned gray after 72 h. Conidia were ellipsoid, hyaline, nonseptate, and formed on botryose heads. They ranged from 5.8 to 9 × 8.1 to 13.7 µm (average 8.6 × 10.2 µm). Black, round, and irregular microsclerotia developed on 7-day-old cultures with an average size of 1.1 × 1.7 mm. Morphological characteristics agree with those described for Botrytis cinerea Pers.:Fr (1). Pathogenicity was tested on 10 12-month-old potted blueberry plants cv. O'Neal by spraying a suspension of 1 × 106 conidia per ml of sterile distilled water. Ten plants used as controls were sprayed with sterile distilled water. Each plant was covered with a transparent polyethylene bag for 48 h and incubated at 20 ± 2°C in humid chambers for 15 days. Lesions similar to those observed in the fields developed after 4 days and asexual fructifications developed after 5 days. The same pathogen was reisolated from the lesions, thus completing Koch's postulates. Water-treated plants remained symptomless. To our knowledge, this is the first report of a disease caused by B. cinerea on blueberry in Buenos Aires, Córdoba, and Entre Ríos provinces of Argentina. References: (1) M. V. Ellis and J. M. Waller. Sclerotinia fuckeliana (conidial state: Botrytis cinerea) No. 431 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1974.

Clinical characteristics of type II diabetic subjects consuming high versus low carbohydrate diets in Mexico City and San Antonio, Texas.

OBJECTIVE: To compare the clinical status of type II diabetic subjects identified in two population-based surveys, one performed in Mexico City, Mexico and the other in San Antonio, Texas. RESEARCH DESIGN AND METHODS: In a low income area of Mexico City, 3,517 age-eligible (35-64 years of age) individuals were randomly selected of whom 3,319 were interviewed at home and 2,198 were examined in a clinic (response rates 62.5%). In San Antonio, 2,357 similarly aged low-income Mexican Americans were randomly selected of whom 2,076 were interviewed at home and 1,511 were examined (response rate 64.1%). Oral glucose tolerance tests were performed at both sites and diabetes was diagnosed according to the World Health Organization (WHO) criteria. In Mexico City, 288 type II diabetic individuals were identified, and 255 were identified in San Antonio. The following variables were measured: height, weight, subscapular and triceps skinfolds, waist-to-hip circumference ratios (WHR), systolic and diastolic blood pressure (random 0 sphygmomanometer), fasting and 2-h postglucose load glucose and insulin concentrations, and fasting total-cholesterol, HDL-cholesterol, and triglyceride (TG) levels. A food frequency questionnaire was used to estimate total calories and the percentage of calories derived from protein, fat, and carbohydrate. Only type II diabetic patients were included in the analyses. Age-adjustment was performed by analysis of covariance for continuous variables and by the Mantel-Haenszel procedure for discrete variables. RESULTS: The mean age, the percentage newly diagnosed cases, and the percentage of males were similar in both sites. The percentage of diabetic patients treated with oral agents was significantly higher in Mexico City (56.9 vs. 72.7% in San Antonio and Mexico City, respectively, P < 0.001), whereas the percentage treated with insulin was significantly higher in San Antonio (18.8 vs. 2.1% for San Antonio and Mexico City, respectively, P < 0.001). A significant difference was observed in the percentage of calories derived from carbohydrate (61.7-63.2 vs. 47.1-47.5% for Mexico City and San Antonio, respectively, P < 0.001) and fat (18.4-20.0 and 30.1-33.0% for Mexico City and San Antonio, respectively, P < 0.001). Body mass index (BMI) was higher in San Antonio (27.6-30.4 vs. 30.2-32.9% for Mexico City and San Antonio, respectively, P < 0.05). Total serum cholesterol was similar at both sites. HDL cholesterol, however, was lower in Mexico City, both in newly and in previously diagnosed patients (30.5-35.8 vs. 39.6-43.3 mg/dl in Mexico City and San Antonio, respectively, P < 0.001). TG levels were higher in Mexico City patients (187-249 vs. 167-179 mg/dl in Mexico City and San Antonio, respectively, P < 0.001). The association between diabetes and the anthropometric and metabolic variables was similar in Mexico City and San Antonio with the following exceptions: Diabetes in Mexico City was associated with less of an elevation in BMI, WHR, and fasting insulin concentration and less of a reduction in the 2-h postoral glucose load insulin concentration compared with diabetes in San Antonio. In addition, although diabetes was associated with a lower HDL in San Antonio subjects, no association appeared between diabetes and HDL in Mexico City subjects. CONCLUSIONS: Diabetic subjects in Mexico City were more likely to be treated with oral agents and less likely to be treated with insulin compared with San Antonio patients. Previously diagnosed diabetic subjects in San Antonio had higher BMIs than diabetic subjects in Mexico City. Diabetic subjects in Mexico City ate less fat but more carbohydrate than those in San Antonio. TG levels were higher and HDL-cholesterol levels were lower in Mexico City diabetic subjects compared with those in San Antonio. San Antonio diabetic subjects had lower HDL levels than nondiabetic subjects but, in Mexico City, HDL levels were similar in diabetic subjects and nondiabetic subjects...

Congenital perilymphatic fistula and associated middle ear abnormalities.

To specifically determine the frequency and type of middle ear abnormalities associated with perilymphatic fistula (PLF), a retrospective chart review was performed of 94 patients (117 ears) who underwent exploratory tympanotomy for PLF from 1980 to 1989. Of the 117 ears explored, 80 (68.4%) had a PLF, and in 65 (81.3%) of these ears, a middle ear malformation was associated with the PLF. Of these 65 ears in which a congenital middle ear abnormality was observed, a malformed stapes was the most common abnormality seen (39 ears, 60%), followed by a deformed round window (20 ears, 30.8%), a deformed incus (11 ears, 16.9%), and a deformed promontory (2 ears, 3%). Often these malformations coexisted amongst themselves or with inner ear abnormalities. Sixteen children (25 ears) had an inner ear malformation identified on computed tomography (CT); all of these children had a PLF found at the time of surgery. This study demonstrated that 86.3% of the ears found to have a PLF had a deformity of the middle ear, inner ear, or both. A malformation of the stapes, most frequently identified as a deformity of its superstructure (and presumably also the anterior footplate), was the most common congenital middle ear abnormality found to be associated with PLF in children.

[Pattern of antimicrobial susceptibility in uropathogen isolates from children]. / Patrón de sensibilidad a antimicrobianos en uropatógenos aislados en niños.

The bacteriological profile in children with culture-positive bacteriuria was analyzed during a 5-year period. Escherichia coli was the most common cause of urinary tract infections (57%), followed by Streptococcus faecalis (11%), Pseudomonas aeruginosa (8%), and Proteus mirabilis (6%). Results of antimicrobial susceptibility testing indicated that cephalosporins (first, second and third generation) and nitrofurantoin are the best empirical oral treatment for urinary infections in children. Fosfomycin is a valid option in some cases. In hospitalized children treatment must be initiated with a third-generation cephalosporin, and gentamicin can be added in severely ill inpatients. These treatments can be modified when microbiological results become available.

First Report of Nectria haematococca Causing Wilt of Olive Plants in Argentina.

Death of 8- to 12-month-old olive plants (Olea europaea L. 'Arbequina', 'Arauco', and 'Picual') has been observed since 1998 in northwestern Argentina. No mycelium or perithecium was observed when examining rotting roots of greenhouse-collected plants. Root segments of diseased plants were plated on potato dextrose agar. Cultures developed a white mycelium after 2 to 3 days, producing microconidia, macroconidia, and chlamydospores identified as Fusarium solani (1). After 15 days of incubation at 23 ± 2°C, reddish perithecia developed infrequently on root segments and adjacent substratum. Single-septate ascospores were hyaline and turned light brown with longitudinal striations at maturity. Microscopic measurements agreed with Nectria haematococca (1). To conduct Koch's postulates (two experiments, two treatments including inoculated and controls, 10 replicates per treatment), young rooted cuttings (6- to 12-month-old) were transferred to pots with a soilless mix and F. solani-colonized oat grains (10:1 vol/vol) and placed in growth chamber (25 to 28°C). First symptoms of the disease were leaf drooping and apex bending after 5 days. At approximately 9 days, leaves turned brownish, developed wilting from the tip downward, and plant death. Controls remained healthy. The fungus was reisolated, and perithecia of N. haematococca developed. F. solani has been reported causing wilt and sudden death in olive previously (2,3). To our knowledge, this is the first report of perithecial development associated with F. solani on olive. References: (1) C. Booth. The Genus Fusarium. CAB International, Wallingford, UK, 1971. (2) R. L. Munjal et al. Studies on diseases of olive in himachal pradesh. Pages 437-440 In: Improvement of Forest Biomass. Symposium Proceedings. Indian Society of Tree Scientists. P. K. Kosla, ed. Sdan, India, 1982. (3) B. A. Pérez et al. (Abstr.) Phytopathology 91 (suppl):S71, 2001.

First Report of Dollar Spot Caused by Sclerotinia homoeocarpa on Agrostis stolonifera in Argentina.

During the summer of 2000, circular, yellow-to-brown, blighted, 2- to 4-cm-diameter patches were observed on creeping bentgrass (Agrostis stolonifera) putting greens (cv. Pennlinks) maintained at a 4- to 5-mm height on a golf course in Pilar (Buenos Aires, Argentina). Symptomatic leaves had transverse chlorotic bands that sometimes extended to the tip with brown lesions inside the bands. A fungus was isolated from symptomatic tissue after surface sterilization with 2% bleach for 1 min and plating on 2% potato dextrose agar (PDA). The mycelium was fluffy and white. The culture turned olive to brown and developed black stromata on the lower side of the plate base after 2 weeks. Pathogenicity tests were performed on 2-month-old healthy plants of A. stolonifera (cv. Crenshaw) grown in sterilized sand. Recently cut, 14-mm-diameter plugs of A. stolonifera were placed in 22- × 17-cm plastic trays filled with a sterilized mixture of 50:50 soil/sand (vol/vol). Plants were maintained at a 7-mm height. Two sources of inoculum were prepared; one was cultured on PDA at 22 to 25°C for 10 days and the other was prepared by incubating in sterilized soil at room temperature for 14 days. Twenty pieces of 1-cm-diameter agar blocks containing mycelium were placed in each plug at the base of the plants. In the infested soil inoculation, 25 g of soil were distributed among the plants on the substrate surface. Control plants were treated with either sterile PDA pieces or noninfested soil. The trays were irrigated with sterilized distilled water, covered with polyethylene bags, and kept in a controlled environment chamber at 25°C with 12 h per day of fluorescent light for 30 days. Leaf chlorosis appeared 7 and 10 days after inoculation for the agar-plug and infested-soil methods, respectively. Leaf necrosis was observed at day 23. Controls remained asymptomatic. The inoculated fungus was reisolated from symptomatic leaf tissue. The pathogen was identified as Sclerotinia homoeocarpa (1,2). To our knowledge, this is the first report of Sclerotinia homoeocarpa causing dollar spot disease on Agrostis stolonifera in Argentina and the first report of a disease on golf courses in our country. References: (1) J. E. M. Mordue. Sclerotinia homoeocarpa. No. 618 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1979. (2) R. W. Smiley. Dollar Spot. Pages 14-16 in: Compendium of Turfgrass Diseases. The American Phytopathological Society, St. Paul, MN, 1983.

Phase-selective image reconstruction of the lungs in small animals using Micro-CT.

Gating in small animal imaging can compensate for artifacts due to physiological motion. This paper presents a strategy for sampling and image reconstruction in the rodent lung using micro-CT. The approach involves rapid sampling of free-breathing mice without any additional hardware to detect respiratory motion. The projection images are analyzed post-acquisition to derive a respiratory signal, which is used to provide weighting factors for each projection that favor a selected phase of the respiration (e.g. end-inspiration or end-expiration) for the reconstruction. Since the sampling cycle and the respiratory cycle are uncorrelated, the sets of projections corresponding to any of the selected respiratory phases do not have a regular angular distribution. This drastically affects the image quality of reconstructions based on simple filtered backprojection. To address this problem, we use an iterative reconstruction algorithm that combines the Simultaneous Algebraic Reconstruction Technique with Total Variation minimization (SART-TV). At each SART-TV iteration, backprojection is performed with a set of weighting factors that favor the desired respiratory phase. To reduce reconstruction time, the algorithm is implemented on a graphics processing unit. The performance of the proposed approach was investigated in simulations and in vivo scans of mice with primary lung cancers imaged with our in-house developed dual tube/detector micro-CT system. We note that if the ECG signal is acquired during sampling, the same approach could be used for phase-selective cardiac imaging.

Genetic analysis of the requirement for flp-2, tadV, and rcpB in Actinobacillus actinomycetemcomitans biofilm formation.

The tad locus of Actinobacillus actinomycetemcomitans encodes a molecular transport system required for tenacious, nonspecific adherence to surfaces and formation of extremely strong biofilms. This locus is dedicated to the biogenesis of Flp pili, which are required for colonization and virulence. We have previously shown that 11 of the 14 tad locus genes are required for adherence and Flp pilus production. Here, we present genetic and phylogenetic analyses of flp-2, tadV, and rcpB genes in biofilm formation. We show that tadV, predicted to encode prepilin peptidase, is required for adherence. In contrast, targeted insertional inactivation of flp-2, a gene closely related to the prepillin gene flp-1, did not abrogate biofilm formation. Expression studies did not detect Flp2-T7 protein under standard laboratory conditions. We present phylogenetic data showing that there is no significant evidence for natural selection in the available flp-2 sequences from A. actinomycetemcomitans, suggesting that flp-2 does not play a significant role in the biology of this organism. Mutants with insertions at the 3' end of rcpB formed biofilms equivalent to wild-type A. actinomycetemcomitans. Surprisingly, 5' end chromosomal insertion mutants in rcpB were obtained only when a wild-type copy of the rcpB gene was provided in trans or when the Tad secretion system was inactivated. Together, our results strongly suggest that A. actinomycetemcomitans rcpB is essential in the context of a functional tad locus. These data show three different phenotypes for the three genes.

Supraestructure maxillectomy and orbital exenteration for treatment of basal cell carcinoma of inferior eyelid: case report and review.

Basal cell carcinoma (BCC) is the most frequent type of skin cancer in humans, with cumulative exposure to ultraviolet radiation as an important risk factor for development of illness such as severe solar burns during childhood or adolescence. BCC is mainly located on sun-exposed sites, head and neck being the areas of more incidences; although nose, eyelids and periorbitary tissue are unfavorable due to cosmetic results that BCC involves. Tumors can be classified as nodular, superficial, micronodular, morphea variety, infiltrating, pigmented, metatypic and fibroepithelioma of Pinkus. Several treatment options such as surgical and nonsurgical are available. The goal of treatment is complete excision of the tumor with preservation of surrounding structures in a way aesthetically acceptable. Mohs' micrographic surgery is the standard treatment for all nonmelanoma skin cancers. Orbital exenteration is also used for treatment of malignancies of ocular tissues, mainly squamous cell carcinoma, sebaceous cell carcinoma and BCC. The tissue beneath the surgical site can be left for second-intention granulation or covered with a cutaneous implant of partial thickness. The case of a 77-year-old patient is presented with BCC of inferior eyelid of 14 years' duration, formerly managed with radiotherapy; however, due to recurrent illness and invasion to the maxillary antrum, he needed supraestructure maxillectomy with left orbital exenteration.

Access as a managed care marketing outcomes measure.

As their position in the health care market diminishes, HMOs are feeling the pinch from the competition. Purchasers of health plans have many more options available today than in the past. Employers can select from single or consolidated health plans, plans offered by coalitions, or plans offered by provider systems. Following closely behind the withstanding issue of controlling costs is quality of care and customer satisfaction. The bad press surrounding managed care is making employers demand assurances that employees will receive the best quality of care their money can buy. To assist in this endeavor managed care companies are focusing more on their customers. To this end marketers use report cards to assess purchaser and enrollee satisfactions, with the hope that if they have a happy customer, s/he will be a loyal one. This paper reviews current marketing strategies of managed care companies and their level of usefulness with respect to sustaining customers and hence market share.