The role of microRNAs in NBS-LRR gene expression and its implications for plant immunity and crop development.

Plants evolved, over millions of years, complex defense systems against pathogens. Once infected, the interaction between pathogen effector molecules and host receptors triggers plant immune responses, which include apoptosis, systemic immune response, among others. An important protein family responsible for pathogen effector recognition is the nucleotide binding site-leucine repeat rich (NBS-LRR) proteins. The NBS-LRR gene family is the largest disease resistance gene class in plants. These proteins are widely distributed in vascular plants and have a complex multigenic cluster distribution in plant genomes. To counteract the genetic load of such a large gene family on fitness cost, plants evolved a mechanism using post transcriptional gene silencing induced by small RNAs, particularly microRNAs. For the NBS-LRR gene family, the small RNAs involved in this silencing mechanism are mainly the microRNA482/2118 superfamily. This suppression mechanism is relieved upon pathogen infection, thus allowing increased NBS-LRR expression and triggering plant immunity. In this review, we will discuss the biogenesis of microRNAs and secondary RNAs involved in this silencing mechanism, biochemical and structural features of NBS-LRR proteins in response to pathogen effectors and the evolution of microRNA-based silencing mechanism with a focus on the miR482/2118 family. Furthermore, the biotechnological manipulation of microRNA expression, using both transgenic or genome editing approaches to improve cultivated plants will be discussed, with a focus on the miR482/2118 family in soybean.

Barriers to patient recruitment in a poststroke neurorehabilitation multicenter trial in Brazil.

There is a high demand for stroke rehabilitation in the Brazilian public health system, but most studies that have addressed rehabilitation for unilateral spatial neglect (USN) after stroke have been performed in high-income countries. Therefore, the aim of this study was to analyze USN patient recruitment in a multicenter noninvasive brain stimulation clinical trial performed in Brazil and to provide study design recommendations for future studies. We evaluated the reasons for exclusion of patients from a multicenter, randomized, double-blinded clinical trial of rehabilitation of USN patients after stroke. Clinical and demographic variables were compared between the included and excluded patients. A descriptive statistical analysis was performed. Only 173 of the 1953 potential neglect patients (8.8%) passed the initial screening. After screening evaluation, 87/173 patients (50.3%) were excluded for clinical reasons. Cognitive impairment led to the exclusion of 21/87 patients (24.1%). Low socioeconomic status led to the exclusion of 37/173 patients (21.4%). Difficulty obtaining transportation to access treatment was the most common reason for their exclusion (16/37 patients, 43.3%). The analyzed Brazilian institutions have potential for conducting studies of USN. The recruitment of stroke survivors with USN was restricted by the study design and limited financial support. A history of cognitive impairment, intracranial stenting or craniectomy, and lack of transportation were the most common barriers to participating in a multicenter noninvasive brain stimulation trial among patients with USN after stroke.

Barriers to patient recruitment in a poststroke neurorehabilitation multicenter trial in Brazil

There is a high demand for stroke rehabilitation in the Brazilian public health system, but most studies that have addressed rehabilitation for unilateral spatial neglect (USN) after stroke have been performed in high-income countries. Therefore, the aim of this study was to analyze USN patient recruitment in a multicenter noninvasive brain stimulation clinical trial performed in Brazil and to provide study design recommendations for future studies. We evaluated the reasons for exclusion of patients from a multicenter, randomized, double-blinded clinical trial of rehabilitation of USN patients after stroke. Clinical and demographic variables were compared between the included and excluded patients. A descriptive statistical analysis was performed. Only 173 of the 1953 potential neglect patients (8.8%) passed the initial screening. After screening evaluation, 87/173 patients (50.3%) were excluded for clinical reasons. Cognitive impairment led to the exclusion of 21/87 patients (24.1%). Low socioeconomic status led to the exclusion of 37/173 patients (21.4%). Difficulty obtaining transportation to access treatment was the most common reason for their exclusion (16/37 patients, 43.3%). The analyzed Brazilian institutions have potential for conducting studies of USN. The recruitment of stroke survivors with USN was restricted by the study design and limited financial support. A history of cognitive impairment, intracranial stenting or craniectomy, and lack of transportation were the most common barriers to participating in a multicenter noninvasive brain stimulation trial among patients with USN after stroke.

Natural Enemies of the Harrisia Cactus Mealybug and Other Hypogeococcus Species (Hemiptera: Pseudococcidae) in Puerto Rico: Identification and Taxonomic Notes on Primary and Secondary Parasitoids.

Primary and secondary parasitoids of Hypogeococcus spp. (Hemiptera: Pseudococcidae) on Cactaceae, Amaranthaceae, and Portulacaceae were reported from field surveys conducted in Puerto Rico, USA, since 2009. Resident natural enemies of the Harrisia cactus mealybug (HCM), Hypogeococcus sp., which is devastating native cacti in Puerto Rico and threatening cacti in the adjacent Caribbean islands, were documented prior to the planned introductions of non-native biological control agents from South America against this pest. Two species of Encyrtidae (Hymenoptera: Chalcidoidea) were recorded as primary parasitoids of the HCM on Cactaceae: Leptomastidea hypogeococci Triapitsyn, which was the most common, and Anagyrus quilmes Triapitsyn, Logarzo & Aguirre. Another primary parasitoid, Acerophagus sp. near nubilipennis Dozier (Encyrtidae), was also reared from a different Hypogeococcus sp. on Amaranthaceae and Portulacaceae. Hyperparasitoids, usually via L. hypogeococci, included Prochiloneurus narendrani Noyes & Triapitsyn (Encyrtidae), Pachyneuron sp. (Hymenoptera: Pteromalidae), Chartocerus elongatus (Girault), and Chartocerus niger (Ashmead) (Hymenoptera: Signiphoridae). Cheiloneurus chrisiantorresi Triapitsyn sp. n. (Encyrtidae) is newly described and illustrated; its status as a suspect hyperparasitoid of Hypogeococcus sp. on Amaranthaceae and Portulacaceae, along with that of Cheiloneurus sp., will need to be verified. The recorded HCM predators include one species of Chamaemyiidae (Diptera), Leucopina bella (Loew), and three species of Coccinellidae (Coleoptera): Cryptolaemus montrouzieri Mulsant, Decadiomus austrinus (Gordon), and Scymnobius flavifrons (Melsheimer).

Evaluation of the cryosurgery for treatment of squamous cell carcinoma in cats / Avaliação da criocirurgia no tratamento do carcinoma de células escamosas em gatos

The cryosurgery is a very useful therapy for the treatment of a variety of neoplastic and non-neoplastic processes. Nevertheless, it is still poorly described as an option for the treatment of specific cutaneous neoplasms, such as squamous cell carcinoma. The purpose of the present study was to analyze the clinical response of cryosurgery for the treatment of squamous cell carcinoma in cats. For this 13 squamous cell carcinoma lesions were selected in 11 cats, diagnosed through citopathological and/or histopathological examinations. The lesions were frozen using liquid nitrogen spray, and the evaluations were performed in the moment of freeze and approximately every 15 days until the wound was completely healed. The response of cryosurgery was considered complete with tumoral remission on 38.5% of the cats, and partial on 46.1%. The main complications included crusting and nostril stenosis. The presented results suggested that cryosurgery is effective and may be a viable option for the treatment of squamous cell carcinoma in cats. The effectiveness of the therapy; however, depends on the correct selection of the candidates for cryosurgery based on the lesion size, and the attendance to some criteria, such as the freezing time and post-operative care.(AU)

A criocirurgia tem se mostrado uma terapia de grande auxílio no tratamento de uma variedade de processos neoplásicos e não neoplásicos. Apesar disso, ainda é pouco descrita a utilização da criocirurgia como opção de tratamento de neoplasias cutâneas específicas, como o carcinoma de células escamosas. Objetivou-se com o presente estudo analisar a resposta clínica da criocirurgia para o tratamento do carcinoma de células escamosas em gatos. Para isso, foram selecionadas 13 lesões do tipo carcinoma de células escamosas em 11 gatos, diagnosticadas por meio de análise citopatológica e/ou histopatológica. As lesões foram congeladas por meio de nitrogênio líquido na forma de spray, e as avaliações realizadas no momento do congelamento e aproximadamente a cada 15 dias, até a completa cicatrização da ferida. A resposta da criocirurgia foi considerada completa, com remissão tumoral em 38,5% dos gatos e parcial em 46,1% destes. As principais complicações verificadas incluíram a formação de crostas e estenose de narina. Os resultados apresentados no presente estudo sugerem que a criocirurgia é efetiva e pode ser uma opção viável para o tratamento do carcinoma de células escamosas em gatos. O sucesso do tratamento, entretanto, depende da correta seleção dos candidatos à técnica quanto ao tamanho da lesão, e do atendimento a alguns critérios, como o tempo de congelamento e os cuidados pós-operatórios.(AU)

Evaluation of the cryosurgery for treatment of squamous cell carcinoma in cats / Avaliação da criocirurgia no tratamento do carcinoma de células escamosas em gatos

The cryosurgery is a very useful therapy for the treatment of a variety of neoplastic and non-neoplastic processes. Nevertheless, it is still poorly described as an option for the treatment of specific cutaneous neoplasms, such as squamous cell carcinoma. The purpose of the present study was to analyze the clinical response of cryosurgery for the treatment of squamous cell carcinoma in cats. For this 13 squamous cell carcinoma lesions were selected in 11 cats, diagnosed through citopathological and/or histopathological examinations. The lesions were frozen using liquid nitrogen spray, and the evaluations were performed in the moment of freeze and approximately every 15 days until the wound was completely healed. The response of cryosurgery was considered complete with tumoral remission on 38.5% of the cats, and partial on 46.1%. The main complications included crusting and nostril stenosis. The presented results suggested that cryosurgery is effective and may be a viable option for the treatment of squamous cell carcinoma in cats. The effectiveness of the therapy; however, depends on the correct selection of the candidates for cryosurgery based on the lesion size, and the attendance to some criteria, such as the freezing time and post-operative care.(AU)

A criocirurgia tem se mostrado uma terapia de grande auxílio no tratamento de uma variedade de processos neoplásicos e não neoplásicos. Apesar disso, ainda é pouco descrita a utilização da criocirurgia como opção de tratamento de neoplasias cutâneas específicas, como o carcinoma de células escamosas. Objetivou-se com o presente estudo analisar a resposta clínica da criocirurgia para o tratamento do carcinoma de células escamosas em gatos. Para isso, foram selecionadas 13 lesões do tipo carcinoma de células escamosas em 11 gatos, diagnosticadas por meio de análise citopatológica e/ou histopatológica. As lesões foram congeladas por meio de nitrogênio líquido na forma de spray, e as avaliações realizadas no momento do congelamento e aproximadamente a cada 15 dias, até a completa cicatrização da ferida. A resposta da criocirurgia foi considerada completa, com remissão tumoral em 38,5% dos gatos e parcial em 46,1% destes. As principais complicações verificadas incluíram a formação de crostas e estenose de narina. Os resultados apresentados no presente estudo sugerem que a criocirurgia é efetiva e pode ser uma opção viável para o tratamento do carcinoma de células escamosas em gatos. O sucesso do tratamento, entretanto, depende da correta seleção dos candidatos à técnica quanto ao tamanho da lesão, e do atendimento a alguns critérios, como o tempo de congelamento e os cuidados pós-operatórios.(AU)

Use of inlet radiographs in the assessment of reduction after the surgical treatment of developmental dysplasia of the hip.

AIMS: This pilot study aimed to evaluate prospectively the use of inlet radiographs of the hip as an alternative method of the assessment of reduction after the surgical treatment of developmental dysplasia of the hip (DDH). PATIENTS AND METHODS: The children in this study underwent surgery between January 2013 and January 2015. All had inlet radiographs and CT scans post-operatively. Data were analysed by determining inter-observer reliability and intra-observer reproducibility, using the kappa value (K). Differences were settled by discussion between the two observers until a consensus was reached. The sensitivity and specificity of the radiographic and CT results were compared. A total of 26 radiographs were obtained from 23 children, with a mean age of 2.38 years (one to five). RESULTS: Similar high levels of intra- and inter-observer agreement were observed (K = 0.834, 95% confidence interval (CI)). There was a high agreement between the radiographic and CT results (K = 0.834, 5% CI), with excellent sensitivity and a specificity of 95.5%. CONCLUSION: These results suggest that inlet radiographs may be a reliable method of assessing the reduction of the hip after the surgical treatment of DDH. Cite this article: Bone Joint J 2017;99-B:697-701.

Comparison of Transpulmonary Thermodilution and Calibrated Pulse Contour Analysis with Pulmonary Artery Thermodilution Cardiac Output Measurements in Anesthetized Dogs.

BACKGROUND: Transpulmonary thermodilution (TPTDCO ) and calibrated pulse contour analysis (PCACO ) are alternatives to pulmonary artery thermodilution cardiac output (PATDCO ) measurement. HYPOTHESIS: Ten mL of ice-cold thermal indicator (TI10 ) would improve the agreement and trending ability between TPTDCO and PATDCO compared to 5 mL of indicator (TI5 ) (Phase-1). The agreement and TA between PCACO and PATDCO would be poor during changes in systemic vascular resistance (SVR) (Phase-2). ANIMALS: Eight clinically normal dogs (20.8-31.5 kg). METHODS: Prospective, experimental study. Simultaneous TPTDCO and PATDCO (averaged from 3 repetitions) using TI5 and TI10 were obtained during isoflurane anesthesia combined or not with remifentanil or dobutamine (Phase-1). Triplicate PCACO and PATDCO measurements were recorded during phenylephrine-induced vasoconstriction and nitroprusside-induced vasodilation (Phase-2). RESULTS: Mean bias (limits of agreement: LOA) (L/min), percentage bias (PB), and percentage error (PE) were 0.62 (-0.11 to 1.35), 16%, and 19% for TI5 ; and 0.33 (-0.25 to 0.91), 9%, and 16% for TI10 . Mean bias (LOA), PB, and PE were 0.22 (-0.63 to 1.07), 6%, and 23% during phenylephrine; and 2.12 (0.70-3.55), 43%, and 29% during nitroprusside. Mean angular bias (radial LOA) values were 2° (-10° to 14°) and -1° (-9° to 6°) for TI5 and TI10 , respectively (Phase-1), and 38° (5°-71°) (Phase-2). CONCLUSIONS AND CLINICAL IMPORTANCE: Although TI10 slightly improves the agreement and trending ability between TPTDCO and PATDCO in comparison to TI5 , both volumes can be used for TPTDCO in replacement of PATDCO . Vasodilation worsens the agreement between PCACO and PATDCO . Because of PCACO 's poor agreement and trending ability with PATDCO during SVR changes, this method has limited clinical application.

Subclinical pulmonary abnormalities in childhood-onset systemic lupus erythematosus patients.

OBJECTIVE: The aims of this study were to analyze the pulmonary function of childhood-onset systemic lupus erythematosus (cSLE) patients and to identify possible correlations between the high-resolution computed chest tomography (HRCT) score, disease activity, disease cumulative damage, and the participants' quality of life. METHODS: Forty cSLE patients, median age: 14.1 years (range: 7.4-17.9), underwent spirometry and plethysmography. Carbon monoxide diffusing capacity (DLCO), HRCT, disease activity, disease cumulative damage, and quality of life were assessed. RESULTS: Pulmonary abnormalities were evident in 19/40 (47.5%) cSLE patients according to spirometry/DLCO. Forced expired volume in one second (FEV1%) was the parameter most affected (30%). The HRCT showed some abnormality in 22/30 patients (73%), which were minimal in 43%. Signs of airway affects were found in 50%. Twelve patients were hospitalized due to cSLE-related pulmonary complications before the study began (median discharge: 2.1 years earlier). Total lung capacity (TLC%), vital capacity (VC%), forced vital capacity (FVC%), and FEV1% were significantly lower in the group with hospitalization compared to the group without hospitalization (p = 0.0025, p = 0.0022, p = 0.0032, and p = 0.0004, respectively). Of note, DLCO was positively correlated with disease duration (r = +0.4; p = 0.01). The HRCT-score was negatively correlated with FEV1/VC (r = -0.63; p = 0.0002), FEV1 (r = -0.54; p = 0.018), FEF25%-75% (r = -0.67; p < 0.0001), and HRCT-score was positively correlated with resistance (r = +0.49; p = 0.0056). CONCLUSIONS: Almost half of patients with cSLE had subclinical pulmonary abnormalities, especially airway abnormalities. The cSLE-related pulmonary complications seem to determine long-term functional damage.

Potent In Vitro Antiproliferative Synergism of Combinations of Ergosterol Biosynthesis Inhibitors against Leishmania amazonensis.

Leishmaniases comprise a spectrum of diseases caused by protozoan parasites of the Leishmania genus. Treatments available have limited safety and efficacy, high costs, and difficult administration. Thus, there is an urgent need for safer and more-effective therapies. Most trypanosomatids have an essential requirement for ergosterol and other 24-alkyl sterols, which are absent in mammalian cells. In previous studies, we showed that Leishmania amazonensis is highly susceptible to aryl-quinuclidines, such as E5700, which inhibit squalene synthase, and to the azoles itraconazole (ITZ) and posaconazole (POSA), which inhibit C-14α-demethylase. Herein, we investigated the antiproliferative, ultrastructural, and biochemical effects of combinations of E5700 with ITZ and POSA against L. amazonensis. Potent synergistic antiproliferative effects were observed against promastigotes, with fractional inhibitory concentration (FIC) ratios of 0.0525 and 0.0162 for combinations of E5700 plus ITZ and of E5700 plus POSA, respectively. Against intracellular amastigotes, FIC values were 0.175 and 0.1125 for combinations of E5700 plus ITZ and E5700 plus POSA, respectively. Marked alterations of the ultrastructure of promastigotes treated with the combinations were observed, in particular mitochondrial swelling, which was consistent with a reduction of the mitochondrial transmembrane potential, and an increase in the production of reactive oxygen species. We also observed the presence of vacuoles similar to autophagosomes in close association with mitochondria and an increase in the number of lipid bodies. Both growth arrest and ultrastructural/biochemical alterations were strictly associated with the depletion of the 14-desmethyl endogenous sterol pool. These results suggest the possibility of a novel combination therapy for the treatment of leishmaniasis.

Comparative study of histopathology and immunohistochemistry of indefinite round cell cutaneous tumors and characterization of canine lymphoma / Estudo comparativo de histopatologia e imuno-histoquímica de tumores indefinidos de células redondas e caracterização de linfoma canino

With the purpose of shedding light on some doubts in veterinary oncology, the present article intends to compare the results of histopathological and immunohistochemical examinations of unspecific round cell neoplasia, to realize immunophenotyping of canine lymphoma cases, to establish the T or B origin of neoplastic cells, and to determine the degree of proliferation and apoptosis of lymphomas by immunohistochemistry. Of 11 animals presenting immunohistochemical diagnosis of lymphoma, five had been diagnosed as Lymphoma by HE staining of histopathological slides and six had been classified as unspecific round cell neoplasia. All cases submitted to immunohistochemical examination were T-cell lymphomas. There was a positive correlation between cell proliferation and apoptosis. The comparison among histopathological and immunohistochemical results obtained in the cases examined in the present study suggested that immunohistochemistry is essential for the differentiation of round cell neoplasia.

Com o objetivo de sanar algumas dúvidas na área da oncologia veterinária, o presente artigo pretende comparar os resultados dos exames histopatológicos e imuno-histoquímicos de neoplasias de células redondas inespecíficas, realizar imunofenotipagem dos casos de linfoma canino e determinar o grau de proliferação e apoptose de linfomas. Dos 11 animais que apresentaram diagnóstico imuno-histoquímico de linfoma, cinco foram diagnosticados como linfoma por coloração HE das lâminas histopatológicas e seis foram classificados como neoplasia de células redondas inespecíficas. Todos os casos submetidos ao exame imuno-histoquímico foram de linfomas de células-T. Houve uma correlação positiva entre a proliferação celular e apoptose. A comparação entre os resultados histopatológicos e imuno-histoquímicos obtidos nos casos analisados sugeriu que a imuno-histoquímica é essencial para a diferenciação das neoplasias de células redondas.

Comparative study of histopathology and immunohistochemistry of indefinite round cell cutaneous tumors and characterization of canine lymphoma / Estudo comparativo de histopatologia e imuno-histoquímica de tumores indefinidos de células redondas e caracterização de linfoma canino

With the purpose of shedding light on some doubts in veterinary oncology, the present article intends to compare the results of histopathological and immunohistochemical examinations of unspecific round cell neoplasia, to realize immunophenotyping of canine lymphoma cases, to establish the T or B origin of neoplastic cells, and to determine the degree of proliferation and apoptosis of lymphomas by immunohistochemistry. Of 11 animals presenting immunohistochemical diagnosis of lymphoma, five had been diagnosed as Lymphoma by HE staining of histopathological slides and six had been classified as unspecific round cell neoplasia. All cases submitted to immunohistochemical examination were T-cell lymphomas. There was a positive correlation between cell proliferation and apoptosis. The comparison among histopathological and immunohistochemical results obtained in the cases examined in the present study suggested that immunohistochemistry is essential for the differentiation of round cell neoplasia.(AU)

Com o objetivo de sanar algumas dúvidas na área da oncologia veterinária, o presente artigo pretende comparar os resultados dos exames histopatológicos e imuno-histoquímicos de neoplasias de células redondas inespecíficas, realizar imunofenotipagem dos casos de linfoma canino e determinar o grau de proliferação e apoptose de linfomas. Dos 11 animais que apresentaram diagnóstico imuno-histoquímico de linfoma, cinco foram diagnosticados como linfoma por coloração HE das lâminas histopatológicas e seis foram classificados como neoplasia de células redondas inespecíficas. Todos os casos submetidos ao exame imuno-histoquímico foram de linfomas de células-T. Houve uma correlação positiva entre a proliferação celular e apoptose. A comparação entre os resultados histopatológicos e imuno-histoquímicos obtidos nos casos analisados sugeriu que a imuno-histoquímica é essencial para a diferenciação das neoplasias de células redondas.(AU)

Effect of immersion and inoculation in ovo of Lactobacillus spp. in embryonated chicken eggs in the prevention of Salmonella Enteritidis after hatch.

The protection level against Salmonella Enteritidis was evaluated in chickens after in ovo treatment with different species of Lactobacillus spp. inoculated into the air cell or by immersion in broth culture. Two hundred forty embryonated eggs were distributed into 8 groups, corresponding to treatments with Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus salivarius, and control. On d 18 of incubation, 4 groups were inoculated with 0.1 mL of inoculum in the air cell and 4 groups were immersed for 3 min in culture of each treatment. Two days after hatching, 0.5 mL of Salmonella Enteritidis culture was inoculated by the intraesophageal route. On d 5 of life, the chicks were euthanized and the ceca were processed to obtain Salmonella Enteritidis counts. There was no decrease in Salmonella Enteritidis colonization of chick ceca, regardless of treatment or route of administration. Lactobacillus spp. samples used in the treatment showed no probiotic potential in chicks when inoculated in ovo, in relation to Salmonella Enteritidis inhibition in poultry ceca.

First Report of Squash vein yellowing virus Affecting Watermelon and Bitter Gourd in Puerto Rico.

In 2005, symptoms of watermelon vine decline (WVD) were observed on a 200-acre watermelon farm in Santa Isabel, on the south central coast of Puerto Rico. WVD symptoms included leaf curling, mosaics, and internode necrosis. In early growth stages of WVD, reduced vigor and plant stunting occurred. At flowering, symptoms progressed to necrosis and wilting of vines. A 2006 to 2007 survey demonstrated that fungal pathogens were not associated with the presence of WVD symptoms (3,4). By 2006, other watermelon fields were also affected. Field trials in 2007 and 2008 with insect-proof cages and insecticides suggested a role of whiteflies (Bemisia tabaci) in the transmission of a virus (3,4). Here, we report that watermelon and pumpkin plants were successfully infected in Puerto Rico by mechanical inoculation and through B. tabaci transmission assays, similarly to transmissions previously conducted in Florida with Squash vein yellowing virus (SqVYV) (1). In addition, plants of Cucurbita moschata exhibited vein clearing symptoms typical of SqVYV after mechanical inoculation with extracts from watermelon plants with WVD symptoms. In 2011, eight watermelon samples from plants exhibiting WVD syndrome were collected in Guánica, Santa Isabel, Juana Díaz, and Mayagüez, and two Momordica charantia samples were collected from Mayagüez. RNA was extracted from all 10 original samples, as well as from plants that were used in mechanical and vector transmission assays, using RNeasy Plant Mini Kit (Qiagen, Valencia, California), and all samples were found positive for SqVYV by reverse transcription-PCR, using previously described primers and methods (1,2). In all cases, a single ~1-kb PCR fragment was revealed, and PCR fragments from four samples were selected for direct sequencing. All sequences showed high levels (>99%) of nucleotide identity with SqVYV sequences from Florida (JF897989, JF897985, and JF897984). Sequences of the SqVYV CP gene from Puerto Rico were deposited in GenBank under accession numbers KC713961 to KC713964. To our knowledge, this is the first report of SqVYV in Puerto Rico associated with WVD syndrome in cucurbits, and thus has implications for management of viral diseases of watermelon in the Caribbean. This is also the first detection of SqVYV outside of the continental United States in both watermelon and a wild species, M. charantia (bitter gourd). References: (1) S. Adkins et al. Phytopathology 97:145, 2007. (2) S. Adkins et al. Plant Dis. 92:1119, 2008. (3) C. Estévez de Jensen et al. Phytopathology 98:S52, 2008. (4) L. Polanco-Florián. El marchitamiento súbito de la sandía [Citrullus lanatus (Thumb.) Matsum & Nakai]. M.S. thesis, University of Puerto Rico, Mayagüez, PR, 2009.

Tomatidine promotes the inhibition of 24-alkylated sterol biosynthesis and mitochondrial dysfunction in Leishmania amazonensis promastigotes.

Leishmaniasis is a set of clinically distinct infectious diseases caused by Leishmania, a genus of flagellated protozoan parasites, that affects ~12 million people worldwide, with ~2 million new infections annually. Plants are known to produce substances to defend themselves against pathogens and predators. In the genus Lycopersicon, which includes the tomato, L. esculentum, the main antimicrobial compound is the steroidal glycoalkaloid α-tomatine. The loss of the saccharide side-chain of tomatine yields the aglycone tomatidine. In the present study, we investigated the effects of tomatidine on the growth, mitochondrial membrane potential, sterol metabolism, and ultrastructure of Leishmania amazonensis promastigotes. Tomatidine (0·1 to 5 µM) inhibited parasite growth in a dose-dependent manner (IC(50)=124±59 nM). Transmission electron microscopy revealed lesions in the mitochondrial ultrastructure and the presence of large vacuoles and lipid storage bodies in the cytoplasm. These structural changes in the mitochondria were accompanied by an effective loss of mitochondrial membrane potential and a decrease in ATP levels. An analysis of the neutral lipid content revealed a large depletion of endogenous 24-alkylated sterols such as 24-methylene-cholesta-5, 7-dien-3ß-ol (5-dehydroepisterol), with a concomitant accumulation of cholesta-8, 24-dien-3ß-ol (zymosterol), which implied a perturbation in the cellular lipid content. These results are consistent with an inhibition of 24-sterol methyltransferase, an important enzyme responsible for the methylation of sterols at the 24 position, which is an essential step in the production of ergosterol and other 24-methyl sterols.

Spatial distributions of the red palm mite, Raoiella indica (Acari: Tenuipalpidae) on coconut and their implications for development of efficient sampling plans.

The red palm mite (Raoiella indica), an invasive pest of coconut, entered the Western hemisphere in 2004, then rapidly spread through the Caribbean and into Florida, USA. Developing effective sampling methods may aid in the timely detection of the pest in a new area. Studies were conducted to provide and compare intra tree spatial distribution of red palm mite populations on coconut in two different geographical areas, Trinidad and Puerto Rico, recently invaded by the mite. The middle stratum of a palm hosted significantly more mites than fronds from the upper or lower canopy and fronds from the lower stratum, on average, had significantly fewer mites than the two other strata. The mite populations did not vary within a frond. Mite densities on the top section of the pinna had significantly lower mite densities than the two other sections, which were not significantly different from each other. In order to improve future sampling plans for the red palm mite, the data was used to estimate the variance components associated with the various levels of the hierarchical sampling design. Additionally, presence-absence data were used to investigate the probability of no mites being present in a pinna section randomly chosen from a frond inhabited by mites at a certain density. Our results show that the most precise density estimate at the plantation level is to sample one pinna section per tree from as many trees as possible.

Evaluation of in vitro and in vivo adhesion and immunomodulatory effect of Lactobacillus species strains isolated from chickens.

This study aimed to characterize the in vitro and in vivo adhesion and immunomodulatory effect of Lactobacillus strains isolated from chickens. Lactobacillus samples isolated from 65-wk-old birds were identified by PCR; their adhesion was evaluated in vitro via basement membrane-type cell matrix and in vivo through carboxyfluorescein succinimidyl amino ester staining inoculation in 1-d-old birds and duodenum, jejunum, ileum, and cecum collections at 1, 4, 12, and 24 h after inoculation. The 5 best adhesive samples at the in vitro test formed a pool for total IgA and IgG measurement in sera and intestinal fluid. The birds were divided into groups by inoculation scheme: group 1 was treated with a pool of Lactobacillus spp. at 2-d-old and challenged 1 d later with Salmonella Enteritidis and then treated again with a pool of Lactobacillus spp. at 4 d of age; group 2 was treated with a pool of Lactobacillus spp. at 2 and 4 d of age; group 3 was challenged with Salmonella Enteritidis at 3 d of age; and group 4 was a negative control. Collections were taken at 7, 14, 21, 28, and 35 d after the first inoculation. The results suggest that basement membrane matrix use represents an important technique for triage of samples for subsequent in vivo evaluation and that carboxyfluorescein succinimidyl amino ester staining is efficient for identifying this bacterial characteristic. The Lactobacillus-treated groups (1 and 2) presented the highest IgA concentrations at the end of the experiment (12,054.6 and 10,568.4 ng/mL, respectively). The group 2 IgG values in intestinal fluid exceeded those of the other 3 groups (P < 0.05), peaking at 6.419 ng/mL. In most serum collections, the Lactobacillus-treated groups (1 and 2) did not differ significantly in IgG concentrations (P > 0.05), whereas group 3 presented the highest concentration of this antibody. It is concluded that there was greater adhesion of strains in the cecum and an important correlation between in vitro and in vivo results. These results also suggest the immunomodulatory action of Lactobacillus spp. in the chicken.