CETP gene polymorphisms and haplotypes are explanatory variables for HDL cholesterol level in sickle cell disease.

Variations in lipid profile have been observed in sickle cell disease (SCD) and understanding their relationship with disease severity is crucial. This study aimed to investigate the association of polymorphisms of the CETP gene and laboratory markers of disease severity with lipid profile in a pediatric population with SCD. Biochemical and anthropometric analyses and CETP and alpha-thalassemia genotyping were performed. The study included 133 children and adolescents with sickle cell anemia (SCA) or hemoglobin SC disease (SCC), in steady-state. The SCA and no hydroxyurea (no HU) groups had higher values of ApoB, total cholesterol, low-density lipoprotein cholesterol (LDL-C), and non-high-density lipoprotein cholesterol (non-HDL-C) compared to the SCC and HU groups. However, there were no significant differences in ApoA1 and HDL-C levels between the groups based on genotype. Furthermore, the groups with altered levels of ApoA1, HDL-C, and the triglyceride/HDL ratio exhibited lower hemoglobin (Hb) levels and higher white blood cell counts. Hb level was associated to HDL-C levels. Analysis of CETP gene variants showed that the minor alleles of rs3764261 (C>A), rs247616 (C>T), and rs183130 (C>T), as well as the TTA haplotype, are explanatory variables for HDL-C levels. These findings suggested that dyslipidemia in SCD, specifically related to HDL-C levels, may be influenced by individual genetic background. Additionally, further investigation is needed to determine if clinical manifestations are impacted by CETP gene variants.

CETP gene polymorphisms and haplotypes are explanatory variables for HDL cholesterol level in sickle cell disease

Variations in lipid profile have been observed in sickle cell disease (SCD) and understanding their relationship with disease severity is crucial. This study aimed to investigate the association of polymorphisms of the CETP gene and laboratory markers of disease severity with lipid profile in a pediatric population with SCD. Biochemical and anthropometric analyses and CETP and alpha-thalassemia genotyping were performed. The study included 133 children and adolescents with sickle cell anemia (SCA) or hemoglobin SC disease (SCC), in steady-state. The SCA and no hydroxyurea (no HU) groups had higher values of ApoB, total cholesterol, low-density lipoprotein cholesterol (LDL-C), and non-high-density lipoprotein cholesterol (non-HDL-C) compared to the SCC and HU groups. However, there were no significant differences in ApoA1 and HDL-C levels between the groups based on genotype. Furthermore, the groups with altered levels of ApoA1, HDL-C, and the triglyceride/HDL ratio exhibited lower hemoglobin (Hb) levels and higher white blood cell counts. Hb level was associated to HDL-C levels. Analysis of CETP gene variants showed that the minor alleles of rs3764261 (C>A), rs247616 (C>T), and rs183130 (C>T), as well as the TTA haplotype, are explanatory variables for HDL-C levels. These findings suggested that dyslipidemia in SCD, specifically related to HDL-C levels, may be influenced by individual genetic background. Additionally, further investigation is needed to determine if clinical manifestations are impacted by CETP gene variants.

First Report of Phytophthora megasperma Causing Decline and Death on Celtis australis in Italy.

European hackberry (Celtis australis L.) is a popular shade tree mainly planted in southern Europe and known to be tolerant to dry and poor soils. In early autumn 2013, hackberry plants grown in soil in a commercial nursery located in the floodplain in Umbria region showed symptoms of wilting, dieback, and death. Up to 100% of the canopy was affected, and over 60% of the plants were symptomatic or dead. A Phytophthora species was consistently isolated from symptomatic 6-year-old plants by plating small pieces of collar and root tissues, cut from the margin of dark-brown necrotic lesions, onto P5ARPH selective medium (4). Pure cultures were obtained by single-hyphal transfers on potato dextrose agar (PDA). Sporangia, produced on pepper seeds in soil extract solution (3), were nonpapillate and noncaducous, measuring 34.0 to 85.0 × 22.0 to 50.0 µm. Oospores had an average diameter of 44 µm with mostly paragynous antheridia. On the basis of morphological features, the isolates were identified as P. megasperma Drech. (2). The identity was confirmed by sequencing the cytochrome c oxidase subunit II (Cox II) (5), which gave 100% identity with P. megasperma sequences available in GenBank (GU222070), and by sequencing the internal transcribed spacer (ITS) using the universal primers ITS4 and ITS6, which gave 99% identity with the AF266794 sequence from Cooke et al. (1). The sequences of one isolate (AB239) were deposited in the European Nucleotide Archive (ENA) with accession numbers HG973451 and HG973450 for Cox II and ITS, respectively. Pathogenicity tests were conducted in the greenhouse with isolate AB239 on eight 2-year-old potted European hackberry plants. Mycelial plugs (5 mm diameter) cut from the margins of actively growing 8-day-old cultures on PDA were inserted through the epidermis to the phloem at the collar level. Two plants were used as controls and treated as described above except that sterile PDA plugs replaced the inoculum. Inoculated plants were kept for 4 weeks in a greenhouse at 24 ± 2°C. During that period, inoculated plants showed wilting symptoms similar to those observed in the field. Lesions were evident at all the inoculation points progressing downward to the roots. Colonies of Phytophthora were isolated from the margins of lesions and identified as P. megasperma, thus fulfilling Koch's postulates. Controls remained symptomless. P. megasperma taxonomy is rather complex since it embraces different subgroups, including host specialized forms (formae speciales), some of which are recognized as biological species. Based on morphological and molecular data presented here, the Phytophthora isolates from hackberry belong to P. megasperma sensu stricto, which is included in the "pathogenic to a broad range of hosts" (BHR) group (1). This pathogen is rather polyphagous, attacking mainly fruit and ornamental woody plants, commonly Prunus spp., Malus spp., and Actinidia deliciosa. Like other homothallic Phytophthora species, it is particularly dangerous due to its abundant production of thick-walled resting oospores that enable long-term survival in the soil. To our knowledge this is the first report of P. megasperma sensu stricto (1) on C. australis and its family Ulmaceae/Cannabaceae. References: (1) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) D. C. Erwin and O. K. Ribeiro, American Phytopathological Society, St. Paul, MN, 1996. (3) E. Ilieva et al. Eur. J. Plant Path. 101:623, 1995. (4) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986. (5) F. N. Martin and P. W. Tooley. Mycologia 95:269, 2003.

First Report of Phytophthora hydropathica Causing Wilting and Shoot Dieback on Viburnum in Italy.

The genus Viburnum comprises over 150 species of shrubs and small trees such as Laurustinus (Viburnum tinus L.), which is one of the most widely used ornamental plants in private and public gardens. Furthermore, it commonly forms stands of natural woodland in the Mediterranean area. In autumn 2012, a survey was conducted to determine the presence of Phytophthora ramorum on Viburnum in commercial nurseries in the Latium region where wilting, dieback, and death of twigs were observed on 30% of the Laurustinus plants. A Phytophthora species was consistently recovered from soil rich in feeder roots from potted Laurustinus plants showing symptoms. Soil samples were baited with rhododendron leaves. Small pieces of leaf tissue cut from the margin of lesions were plated on P5ARPH selective medium (4). Pure cultures, obtained by single-hypha transfers on potato dextrose agar (PDA), were petaloid. Sporangia formation was induced on pepper seeds (3). Sporangia were almost spherical, ovoid or obpyriform, non-papillate and non-caducous, measuring 36.6 to 71.4 × 33.4 to 48.3 µm (average 53.3 × 37.4 µm) with a length/width ratio of 1.4. Chlamydospores were terminal and 25.2 to 37.9 µm in diameter. Isolates were considered heterothallic because they did not produce gametangia in culture or on the host. All isolates examined had 30 to 35°C as optimum temperatures. Based on these morphological characteristics, the isolates were identified as Phytophthora hydropathica (2). Morphological identification was confirmed by internal transcribed spacer (ITS), and mitochondrial partial cytochrome oxidase subunit 2 (CoxII) with BLAST analysis in the NCBI database revealing 99% identity with ITS and 100% identity with CoxII. The sequences of the three isolates AB234, AB235, and AB236 were deposited in European Nucleotide Archive (ENA) with the accession nos. HG934148, HG934149, and HG934150 for ITS and HG934151, HG934152, and HG934153 for CoxII, respectively. Pathogenicity tests were conducted in the greenhouse on a total of six 1-year-old shoots cut from V. tinus plants with two inoculation points each. Mycelial plugs cut from the margins of actively growing 8-day-old cultures on PDA were inserted through the epidermis into the phloem. Controls were treated as described above except that sterile PDA plugs replaced the inoculum. Shoots were incubated in test tubes with sterile water in the dark at 24 ± 2°C. After 2 weeks, lesions were evident at the inoculation points and symptoms were similar to those caused by natural infection. P. hydropathica was consistently re-isolated from the margin of lesions, while controls remained symptomless. In the United States in 2008, P. hydropathica was described as spreading from irrigation water to Rhododendron catawbiense and Kalmia latifolia (2). This pathogen can also attack several other horticultural crops (1), but to our knowledge, this is the first report of P. hydropathica causing wilting and shoot dieback on V. tinus. References: (1) C. X. Hong et al. Plant Dis. 92:1201, 2008. (2) C. X. Hong et al. Plant Pathol. 59:913, 2010. (3) E. Ilieva et al. Eur. J. Plant Path. 101:623, 1995. (4) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986.

Gene expression profiling in breast cancer.

In recent years, molecular research has translated into remarkable changes of breast cancer diagnostics and therapeutics. Molecular tests such as the 21 gene expression test (Oncotype DX(TM)) and 70 gene microarray test (MammaPrint(®)) have revolutionized the predictive and prognostic tools in the clinic. By stratifying the risk of recurrence for patients, the tests are able to provide clinicians with more information on the treatment outcomes of using chemotherapy, HER2 targeted therapy or endocrine therapy or the combination of the therapies for patients with particular genetic expressions. However, it is still questionable for clinical applications as some areas remain unclear and that the true benefit still needs prospective evaluation. Such studies are under way and are anxiously awaited. In this paper, the limitation of the molecular tests are discussed. As we are moving towards personalized medicine, molecular profiling will not only result in better outcomes but in a certain proportion of patients, likely will spare unnecessary use of cytotoxic compounds and reduce the cost to the health care systems.

First Report of Phytophthora tropicalis on Rhododendron in Italy.

The genus Rhododendron comprises over 1,000 species, which represent many important ornamental shrubs. Microbial isolations were made from Rhododendron catawbiense plants showing symptoms of wilt, dieback, and death of shoots obtained from two nurseries in the Latium region in the late summer of 2012. A Phytophthora species was consistently recovered by plating small pieces of stem and collar tissues, cut from the margin of lesions, on P5ARPH selective medium. Pure cultures were obtained by single-hyphal transfers and they grew in a rosaceous pattern on potato dextrose agar (PDA) at an optimum temperature of 28 to 30°C. Sporangia formation was induced on pepper seeds (3). Sporangia were ellipsoid, fusiform or obpyriform, papillate, occasionally bipapillate, caducous, with a long pedicel (up to 100 µm), and mean dimensions of 45 × 25 µm with a mean length/width ratio of 1.8. Chlamydospores ranged from 25 to 32 µm in diameter. Isolates were considered heterothallic because they did not produce gametangia in vitro or in planta. On the basis of morphological features, the isolates were identified as Phytophthora tropicalis Aragaki & Uchida. Identity was confirmed by sequence comparison in GenBank with 99% homology both for internal transcribed spacer (ITS) and mitochondrial partial COI for cytochrome oxidase subunit 1. The sequences of two isolates AB211 and AB212 were deposited in the European Nucleotide Archive (ENA) with accession nos. HF937577 and HF937578 for ITS, and HF937579 and HF937580 for COI, respectively. Pathogenicity tests were conducted in the greenhouse on a total of six 1-year-old shoots cut from R. catawbiense plants with two inoculation points each. Mycelial plugs cut from the margins of actively growing 8-day-old cultures on PDA were inserted through the epidermis to the phloem. Controls were treated as described above except for inoculation with sterile PDA plugs. Inoculated shoots were incubated in test tubes with sterile water for 1 week in the dark at 26 ± 2°C. Lesions were evident at the inoculation points. P. tropicalis was consistently reisolated from the margin of symptomatic tissues. Control shoots remained symptomless. In Italy, P. tropicalis has been reported on several ornamental species (1) and on apricot trees (4) indicating a broad host range. On the same host it has been reported in Virginia, United States (2). To the best of our knowledge, this is the first report of Phytophthora damage on Rhododendron caused by P. tropicalis in Italy. References: (1) S. O. Cacciola et al. Plant Dis. 90: 680, 2006. (2) C. X. Hong et al. Plant Dis. 90: 525, 2006. (3) E. Ilieva et al. Eur. J. Plant Path. 101: 623, 1995. (4) A. Pane et al. Plant Dis. 93: 844, 2009.

Emerging role of multikinase inhibitors for refractory thyroid cancer.

Thyroid cancer incidence continues to increase, remaining the most common endocrine malignancy. The need for effective systemic therapies combined with high incidence of driver mutations and overexpression of molecular pathways make refractory thyroid cancer an ideal candidate for treatment with novel agents. Multikinase inhibitors have caused a paradigm shift in the treatment of patients with advanced iodine-refractory thyroid cancer. These agents have shown to be the most effective systemic therapy for this disease not only causing prolonged responses but also improving survival. The activity of these agents inhibiting several pathways simultaneously, such as rearranged during transfection protooncogene, mitogen-activated protein kinase, and angiogenesis, can probably explain the effectiveness in controlling the progression of this malignancy. Several of these agents are currently on clinical studies in patients with differentiated and medullary thyroid cancer and most of them are showing promising clinical activity. With the approval of vandetanib for the treatment of medullary thyroid cancer, a new era in the management of this disease has begun. The molecular rationale for the use of these drugs for thyroid cancer is discussed as well as their promising clinical results.

Safety and efficacy of pemetrexed in maintenance therapy of non-small cell lung cancer.

Lung cancer incidence continues to rise and is the number one cause of cancer death in both men and women worldwide with projected 221,130 new cases and 156,940 deaths in the United States in 2011.1 Non-small cell lung cancer (NSCLC) represents more than 85% of the cases with most patients having either locally advanced or metastatic disease at the time of initial diagnosis, and approximately 60%-70% of them have an adenocarcinoma histologic subtype. In the last three years, we have seen several advances in the management of NSCLC, with several factors playing an important role in the treatment decision making process. Maintenance therapy has been added to the algorithm of NSCLC management and Pemetrexed has been studied as single agent or in combination in this setting with recent studies showing safety and improved progression free survival (PFS) and/or overall survival (OS), still the disease for the most part has a dismal outcome. More research work needs to be done to identify which patients truly benefit from these approaches, and to whom we should offer maintenance or switch maintenance vs. close observation.

Novel molecular targeted therapies for refractory thyroid cancer.

The incidence of thyroid cancer continues to increase and this neoplasia remains the most common endocrine malignancy. No effective systemic treatment currently exists for iodine-refractory differentiated or medullary thyroid carcinoma, but recent advances in the pathogenesis of these diseases have revealed key targets that are now being evaluated in the clinical setting. RET (rearranged during transfection)/PTC (papillary thyroid carcinoma) gene rearrangements, B-Raf gene mutations, and vascular endothelial growth factor receptor 2 (VEGFR-2) angiogenesis pathways are some of the known genetic alterations playing a crucial role in the development of thyroid cancer. Several novel agents have demonstrated promising responses. Of the treatments studied, multi-kinase inhibitors such as axitinib, sorafenib, motesanib, and XL-184 have shown to be the most effective by inducing clinical responses and stabilizing the disease process. Randomized clinical trials are currently evaluating these agents, results that may soon change the management of thyroid cancer.

Mechanisms governing the responses to anthracnose pathogen in Juglans spp.

Juglans nigra and Juglans regia are two highly economically important species for wood and fruit production that are susceptible to anthracnose caused by Gnomonia leptostyla. The identification of genotypes resistant to anthracnose could represent a valid alternative to agronomic and chemical management. In this study, we analyzed 72 walnut genotypes that showed a variety of resistance phenotypes in response to natural infection. According to the disease severity rating and microsatellite fingerprinting analysis, these genotypes were divided into three main groups: (40) J. nigra resistant, (1) J. nigra susceptible, and (31) J. regia susceptible. Data on leaf emergence rates and analysis of in vivo pathogenicity indicated that the incidence of anthracnose disease in the field might be partially conditioned by two key factors: the age and/or availability of susceptible leaves during the primary infection of fungus (avoidance by late flushing) and partial host resistance. NBS profiling approach, based on PCR amplification with an adapter primer for an adapter matching a restriction enzyme site and a degenerate primer targeting the conserved motifs present in the NBS domain of NBS-LRR genes, was applied. The results revealed the presence of a candidate marker that correlated to a reduction in anthracnose incidence in 72 walnut genotypes.

First Report of Phytophthora megasperma Associated with Decline and Death of Common Walnut Trees in Italy.

Common walnut (Juglans regia L.) is an important nut crop in Italy, which is the fifth largest producer of walnut in Europe. In recent years, walnut decline and subsequent death has increased in many Italian commercial orchards. In the summer of 2010, several declining trees were present in waterlogged area of a walnut orchard located in the Veneto region. Symptoms included sparse foliage, wilting, and shoot and branch dieback. By the next year, a larger area of about 1 ha with 20% of dead trees was present, and soil/root samples were subjected to azalea leaf baiting and successively cultured on PARBH medium (3). Isolates were identified as Phytophthora megasperma based on morphological characteristics (2) and DNA sequence analysis. Sporangia were 35.0 to 62.0 × 12.0 to 30.0 µm, nonpapillate, and noncaducous when produced in soil extract solution. Oogonia had an average diameter of 36 µm with mostly paragynous antheridia. Identity was confirmed by sequence comparison in NCBI database with 99% and 100% identity for internal transcribed spacer (ITS) and mitochondrial partial COI (4) for cytochrome oxidase subunit 1, respectively. The sequences of the isolate AB199 were deposited in GenBank with the accession nos. HE805270 and HE805269 for ITS and COI, respectively. Pathogenicity tests were conducted in the greenhouse on six 1-year-old walnut shoots with two inoculation points each. Mycelial plugs cut from the margins of actively growing 8-day-old cultures on PDA were inserted through the epidermis to the phloem. Controls were treated as described above except for inoculation with sterile PDA plugs. After inoculation, shoots were incubated in test tubes with sterile water for 1 week in the dark at 22 ± 2°C. Lesions were evident at the inoculation points. P. megasperma was consistently reisolated from the margin of symptomatic tissues. Controls remained symptomless. P. megasperma is a polyphagous, ubiquitarious Phytophthora species that attacks many crops and fruit species including walnut. Although several other species of Phytophthora have been reported from Italy (1), to the best of our knowledge, this is the first report of Phytophthora decline on common walnut in Italy caused by P. megasperma. References: (1) A. Belisario et al. Acta Hort. 705:401, 2006. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (3) G. C. Papavisas et al. Phytopathology 71:129, 1981. (4) G. P. Robideau et al. Mol. Ecol. Resour. 11:1002, 2011.

Development of targeted therapy for squamous cell carcinomas of the head and neck.

Targeted therapy is a very exciting era in the treatment of squamous cell carcinomas of the head and neck. After adding cetuximab to conventional chemotherapy and radiation therapy, we are strongly considering the role of induction chemotherapy with the addition of docetaxel. At the same time, other new treatments, especially targeted agents and novel combined regimens, are being evaluated in ongoing clinical trials. For example, several trials are attempting to combine docetaxel and cetuximab in chemoradiation or induction settings. However, in the near future we are likely to see a strong presence of targeted agents that have been found to be not only effective, but also less toxic than conventional chemotherapeutic agents. Their toxicity profiles make them eligible for addition to radiation treatment strategies, as well as other chemotherapy agents, or even for replacing these chemotherapy agents. In this article, we are going to review the indications and current role of cetuximab, tyrosine kinase inhibitors (gefitinib and erlotinib), dual inhibitors, IGF receptor inhibitors, as well as other agents that are in development for treatment of head and neck squamous cell carcinomas.

Morphological and molecular analysis of Fusarium lateritium, the cause of gray necrosis of hazelnut fruit in Italy.

Fusarium lateritium is a globally distributed plant pathogen. It was recently reported as the causal agent of nut gray necrosis (NGN) on hazelnut. Isolate characterization within F. lateritium was undertaken to investigate how morphological and molecular diversity was associated with host and geographic origin. Morphological studies combined with inter-simple-sequence repeat (ISSR) analysis, and phylogenetic analyses using translation elongation factor 1α (TEF-1α), ß-tubulin genes, and nuclear ribosomal DNA internal transcribed spacer (ITS) sequences were conducted to resolve relationships among 32 F. lateritium isolates from NGN-affected hazelnut fruit, and 14 from other substrates or 8 from other hosts than hazelnut. Colonies of F. lateritium from hazelnut showed dark grayish-olive differing from the orange-yellow color of all other isolates from other hosts. Generally, isolates from NGN-affected fruit failed to produce sporodochia on carnation leaf agar. The influence of host and substrate on the genetic structure of F. lateritium was supported by ISSR and analyzed with principal coordinates analysis. A relationship between hazelnut and genetic variation was inferred. Phylogenetic analysis of ITS provided limited resolution while TEF-1α and ß-tubulin analyses allowed a clear separation between the European and non-European F. lateritium isolates retrieved from GenBank, regardless of host. Though morphological traits of F. lateritium isolates from hazelnut were generally uniform in defining a typical morphogroup, they were not yet phylogenetically defined. In contrast, the typology related to slimy deep orange cultures, due to spore mass, grouped clearly separated from the other F. lateritium isolates and revealed a congruence between morphology and phylogeny.

Nasopharyngeal carcinoma: alternative treatment options after disease progression.

Nasopharyngeal carcinoma is a rare malignancy with an incidence well under one per 100,000 person-years, except for some geographic areas, such as Asia. The prognosis of nasopharyngeal carcinoma is related to its potential for locoregional invasion and metastatic spread. Radiotherapy alone remains the standard treatment for early stages. However, for locally advanced disease, chemotherapy may offer some benefit as a radiosensitizer while treating microscopic spread disease. Chemoradiotherapy is now the standard treatment for locally advanced and/or node-positive patients. Platinum-based therapy is the preferred regimen for this therapeutic approach. In this review, we discuss all treatment modalities available for nasopharyngeal carcinoma, including the standard of care and what therapeutic options could be available for those patients who progress after the standard treatment has been delivered.

Analysis of cytokine genes polymorphism as markers for inhibitor development in haemophilia A.

Antibodies that block factor VIII (FVIII) activity appear in some haemophilia A patients treated with FVIII replacement therapy and severely impaired treatment. To date, the mechanisms that lead to this immune response are unknown. In this work, haplotypes of cytokine interleukin 10 (IL-10) gene have been associated with the presence of FVIII inhibitors in a group of Brazilian haemophilia A patients. The coexistence of a haplotype defining high IL-10 synthesis and one defining an intermediate production of cytokines is found to be associated with the group of patients who have a history of inhibitor development. Additionally, the coexistence of haplotypes defining high and low IL-10 syntheses is strongly associated with the group of negative inhibitors. These results have shown that the simple association considering only the presence or the absence of a haplotype and the development of inhibitors in haemophilia A is not sufficient. Data obtained in this work sustain the idea that the genetic studies may partly explain why only approximately 25% of haemophilia A patients develop FVIII inhibitors. Other genetic issues and details of the protein replacement therapy should be considered to measure the chances of a patient to develop anti-FVIII antibodies.

First Report of Fusarium semitectum as the Agent of Twig Cankers on Persian (English) Walnut in Italy.

English (Persian) walnut (Juglans regia) is the most widely cultivated walnut species. During the last 10 years, the increment of walnut cultivation in Italy has been accompanied by changes in cultural management. Changes were addressed to develop highly specialized cultivations with intensive pruning, fertilization, irrigation, and chemical treatments. For these reasons, more consideration has been given to the sanitary situation, in particular since 1998 when brown apical necrosis (BAN) was first observed (1). BAN is a fungal complex disease causing fruit drop, in which several Fusarium spp. are involved, among which Fusarium semitectum represents one of the major causal agents (2). From 2005 onward, investigations on sources of inoculum for BAN led to observing the presence of twig cankers on walnut trees cv. Lara located in northern Italy (Po valley). Cankers observed in late spring to summer were usually small (1 to 3 cm long) and mainly occurred on the new growth strongly incited by intensive pruning. Pale orange sporodochia were evident on lesions. Isolations were made from the margins of lesions, and small fragments of tissues (approximately 3 mm) were plated onto potato dextrose agar (PDA) after surface disinfection with 1% NaOCl. Whitish, light brown colonies were consistently obtained. On PDA, the production of fusoid, 1- to 3-celled mesoconidia was abundant. This characteristic was combined with the presence of two-spored polyphialides with a "rabbit-ear" appearance. Three to five septate macroconidia (38 × 4 µm) were produced in sporodochia on carnation leaf agar (CLA). On the basis of morphological characteristics (3), the fungus was identified as F. semitectum Berk. & Ravenel (synonyms F. incarnatum and F. pallidoroseum). Sequence comparison of internal transcribed spacer (ITS) and translation elongation factor 1-alpha (TEF1-α) was used to support the identification. A 99% identity for ITS was obtained with Accession No. AY633745 from Vietnam, while for TEF 1-α, a comparison was not available in GenBank. The sequences of one isolate (ISPaVe1946) were deposited in GenBank (Accession No. FN430680 for ITS and No. FN430737 for TEF 1-α). Pathogenicity tests were conducted outdoors on 1-year-old shoots of J. regia potted plants using ISPaVe1946 single-spored isolate. Mycelial plugs of 5-mm diameter, cut from the margin of PDA actively growing cultures, were placed under the bark and protected with Parafilm to prevent desiccation. Six inoculation points were performed. Controls were inoculated with plain PDA plugs. Within 2 months after inoculation, cankers developed in all inoculated points and were similar to those observed in nature. Controls showed no symptoms. Koch's postulates were fulfilled and the pathogen was constantly reisolated from lesions. To our knowledge, this is the first report of F. semitectum as the causal agent of twig cankers on walnut in Italy. This pathogen was already reported as an agent of canker on walnut in Argentina (4). References: (1) A. Belisario et al. Inf. Agrario 21:51, 1999. (2) A. Belisario et al. Plant Dis. 86:599, 2002. (3) J. F. Leslie et al. The Fusarium Laboratory Manual. Blackwell Publishing Press Ltd, Oxford, UK, 2006. (4) S. Seta et al. Plant Pathol. 53:248, 2004.

First Report of Leaf Blight and Root and Foot Rot of a Strelitzia Caused by Phytophthora nicotianae in Italy.

Bird of paradise, also known as crane flower (Strelitzia reginae Aiton), is a monocotyledonous flowering plant indigenous to South Africa. It is commonly grown and commercialized as an ornamental plant and it is appreciated for its beautiful flowers. In October of 2008, dark leaf spots and leaf blight associated with a severe root and foot rot were observed on several plants of S. reginae grown in a private garden located in Fiumicino, Italy. Small fragments of tissues (approximately 3 mm) collected from the base of leaves and roots and the margins of brown lesions, previously surface disinfected with 0.5% NaOCl, were plated onto potato dextrose agar (PDA) and incubated at 22°C in the dark. White, web-like, slow-growing colonies with coenocytic mycelium and hyphal swellings consistently developed from all plated tissue samples. Sporangia were ovoid or ellipsoid with prominent papillae (including some bipapillate) and frequently caducous with a short stalk. The dimensions of sporangia were 27 to 64 × 23 to 45 µm (average 43 × 35 µm). Chlamydospores were terminal or intercalary and approximately 30 µm in diameter. Isolates were considered heterothallic because they did not produce gametangia in vitro or in planta. On the basis of morphological features, isolates were identified as Phytophthora nicotianae (Breda de Haan). The identity was confirmed by internal transcribed spacer (ITS) sequence comparison in NCBI database with 99% identity with sequences available in GenBank (e.g., EU331089) and with cytochrome c oxidase subunit I (Cox I) with 99% identity with AY564196 by Kroon et al. (2). The sequences of one isolate, AB177, were deposited in GenBank (Accession Nos. FN430681 and FN552051 for ITS and Cox I, respectively). Pathogenicity tests were conducted in the greenhouse on leaves of a 1-year-old potted S. reginae plant by placing 5-mm-diameter mycelial plugs, cut from the margins of 10-day-old actively growing cultures, with mycelium in contact with plant tissues gently wounded with a needle. Controls were treated as described above, except that PDA sterile plugs were used. Plants were misted with water and placed in sealed plastic bags for 48 h. After 10 days, artificially wounded strelitzia leaves showed lesions (approximately 1 cm long). Controls remained symptomless. All inoculated leaves showed the same leaf symptoms as observed on naturally diseased plants. The pathogen was consistently reisolated from lesions. P. nicotianae has been reported as the causal agent of leaf blight and stem, collar, and root rot on several plants (1). It has been reported as an agent of Phytophthora blight on strelitzia in Japan (3). To our knowledge, this is the first report of P. nicotianae on strelitzia in Italy. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) L. P. N. M. Kroon et al. Fungal Genet. Biol. 41:766, 2004. (3) S. Uematsu et al. Ann. Phytopathol. Soc. Jpn. 60:746, 1994.

First Report of Fusarium lateritium as the Agent of Nut Gray Necrosis on Hazelnut in Italy.

Hazelnut (Corylus avellana) is a traditionally cultivated nut species in Italy. Italy is the second largest producer of hazelnut in the world after Turkey. In early summer of 2000, a severe fruit drop (up to 60%) was observed in several hazelnut orchards located in the Latium Region in central Italy. The severity of yield losses led to investigating the etiology of the disease subsequently named nut gray necrosis (NGN) based on symptoms observed on the affected fruit. Symptoms consisted of a brown grayish necrotic spot/patch on the nut shell and bracts, sometimes involving the petiole (1). Isolations of the potential pathogen were from tissue that was sampled starting from bloom of female flowers to fully ripened fruit. Isolations from symptomatic tissue consisted of placing onto potato dextrose agar (PDA) small tissue fragments (approximately 3 mm) cut from the margin of lesions, while asymptomatic material (entire flowers or young fruit) was sectioned into small pieces. All the material was previously surface disinfected with 1% NaOCl. Slow-growing, dark grayish olive colonies were obtained consistently within 14 days of incubation at 20 to 22°C from symptomatic and asymptomatic material. Sporodochia were rarely produced on PDA, but never on carnation leaf agar. Dark grayish olive colonies were assigned to a Fusarium sp. Detached hazelnut fruit exposed to 20 µl of a mycelial suspension (105 CFU/ml) and incubated in a moist chamber at room temperature for 10 days produced orange sporodochia bearing 3 to 5 septate macroconidia (35 × 4 µm). On the basis of morphology, the fungus was identified as Fusarium lateritium Nees. (3,4). The identity was confirmed by internal transcribed spacer rDNA sequence comparison with BBA65248 (GenBank Accession No. AF310980). The sequences of two isolates, ISPaVe1874 and ISPaVe1976, were deposited in GenBank (Accession Nos. FN547420 and FN547445, respectively). Pathogenicity tests were performed in planta by inoculating, with the aforementioned isolates, young to fully formed fruit (approximately 24 mm in diameter) either with a drop of mycelial (106 CFU/ml) or conidial (106 conidia/ml) suspension. Drops were placed between the nut and leafy involucre. Controls were treated with sterile water only. Within 2 weeks after inoculation, a grayish necrosis developed on all the inoculated fruit and was similar to symptoms originally observed in the field. No differences were observed between the two methods of inoculation. On full-sized fruit, lesions extended from the shell to inner tissues. The pathogen was consistently reisolated from lesions. Controls showed no symptoms. To our knowledge, this is the first report of F. lateritium as the causal agent of nut gray necrosis on hazelnut in Italy, and this pathogen has never been reported as an agent of necrosis and drop of hazelnut fruit, but it was previously reported as an agent of twig cankers (2). References: (1) A. Belisario et al. Inf. Agrario 59:71, 2003. (2) A. Belisario et al. Plant Dis. 89:106, 2005. (3) J. F. Leslie et al. The Fusarium Laboratory Manual. Blackwell Publishing Press Ltd, Oxford, UK, 2006. (4) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. Pennsylvania State University Press, 1983.

Second-line therapy for non-small-cell lung cancer.

Carcinogenesis is a complex pathological process induced by abnormalities in the genome, cell-cycle dysregulation, loss of the programmed cell death process, and upregulation of oncogenic pathways associated with proliferation, migration, and survival, among others. Despite recent advances in molecular and tumor biology in non-small-cell lung cancer (NSCLC) and the introduction of several targeted agents, the disease continues to have a dismal survival. Nonetheless, the future looks promising; conventional cytotoxic chemotherapy regimens in combination with targeted agents have shown better response rates and survival than those seen in the past. These targeted agents have the advantage of blocking or inhibiting specific pathways necessary for tumor growth, proliferation, and metastases, without significantly affecting quality of life by having an acceptable toxicity profile. Thus, these novel agents harbor a hope in the treatment of NSCLC and many other malignant diseases when they can be used either in combination with other chemotherapy drugs in several lines of treatment or as a single agent in maintenance therapy until progression of disease, or even more attractively, in combination with other targeted agents themselves. In this review, we discuss second-line treatments for patients who have NSCLC, including targeted agents and their development in this specific setting as part of our armamentarium in lung cancer.

Reactivity profile of anti-factor VIII antibodies with designed synthetic peptides mimicking epitopes of the C2 and a1 domains.

Antibodies (Abs) that block factor VIII (FVIII) activity occur in hemophilia A patients treated with FVIII replacement therapy and severely impair treatment. In this work, we designed and synthesized ten peptides whose sequences are found in putative epitopes at the surface of a1 and C2 domains of the FVIII molecule. These peptides were screened for their ability to inhibit the binding of anti-FVIII Abs from plasmas of hemophilia A patients to FVIII. All peptides were efficient in inhibiting anti-FVIII Abs in plasma from patients with inhibitors, with however different efficiencies. It was found that each tested patient's plasma had a different profile of reactivity with peptides, consistent with an individual anti-FVIII Ab specificity. The profile of recognized peptides was also changing during the treatment of the patients. Three peptides were used in an affinity chromatography assay to attempt to remove anti-FVIII Abs from patients' plasma. Anti-FVIII IgGs were significantly captured by the peptide-Sepharose affinity matrixes as assessed by enzyme-linked immunosorbent assay. However, due to the low level of Abs in the plasma samples, other methods (Chromogenic and Bethesda assays) were not sensitive enough to properly detect the reduction of inhibitors.