Routes of Hendra Virus Excretion in Naturally-Infected Flying-Foxes: Implications for Viral Transmission and Spillover Risk

Pteropid bats or flying-foxes (Chiroptera: Pteropodidae) are the natural host of Hendra virus (HeV) which sporadically causes fatal disease in horses and humans in eastern Australia. While there is strong evidence that urine is an important infectious medium that likely drives bat to bat transmission and bat to horse transmission, there is uncertainty about the relative importance of alternative routes of excretion such as nasal and oral secretions, and faeces. Identifying the potential routes of HeV excretion in flying-foxes is important to effectively mitigate equine exposure risk at the bat-horse interface, and in determining transmission rates in host-pathogen models. The aim of this study was to identify the major routes of HeV excretion in naturally infected flying-foxes, and secondarily, to identify between-species variation in excretion prevalence. A total of 2840 flying-foxes from three of the four Australian mainland species (Pteropus alecto, P. poliocephalus and P. scapulatus) were captured and sampled at multiple roost locations in the eastern states of Queensland and New South Wales between 2012 and 2014. A range of biological samples (urine and serum, and urogenital, nasal, oral and rectal swabs) were collected from anaesthetized bats, and tested for HeV RNA using a qRT-PCR assay targeting the M gene. Forty-two P. alecto (n = 1410) had HeV RNA detected in at least one sample, and yielded a total of 78 positive samples, at an overall detection rate of 1.76% across all samples tested in this species (78/4436). The rate of detection, and the amount of viral RNA, was highest in urine samples (>serum, packed haemocytes >faecal >nasal >oral), identifying urine as the most plausible source of infection for flying-foxes and for horses. Detection in a urine sample was more efficient than detection in urogenital swabs, identifying the former as the preferred diagnostic sample. The detection of HeV RNA in serum is consistent with haematogenous spread, and with hypothesised latency and recrudesence in flying-foxes. There were no detections in P. poliocephalus (n = 1168 animals; n = 2958 samples) or P. scapulatus (n = 262 animals; n = 985 samples), suggesting (consistent with other recent studies) that these species are epidemiologically less important than P. alecto in HeV infection dynamics. The study is unprecedented in terms of the individual animal approach, the large sample size, and the use of a molecular assay to directly determine infection status. These features provide a high level of confidence in the veracity of our findings, and a sound basis from which to more precisely target equine risk mitigation strategies.

Natural Hendra Virus Infection in Flying-Foxes - Tissue Tropism and Risk Factors

Hendra virus (HeV) is a lethal zoonotic agent that emerged in 1994 in Australia. Pteropid bats (flying-foxes) are the natural reservoir. To date, HeV has spilled over from flying-foxes to horses on 51 known occasions, and from infected horses to close-contact humans on seven occasions. We undertook screening of archived bat tissues for HeV by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Tissues were tested from 310 bats including 295 Pteropodiformes and 15 Vespertilioniformes. HeV was detected in 20 individual flying-foxes (6.4%) from various tissues including spleen, kidney, liver, lung, placenta and blood components. Detection was significantly higher in Pteropus Alecto and Pconspicillatus, identifying species as a risk factor for infection. Further, our findings indicate that HeV has a predilection for the spleen, suggesting this organ plays an important role in HeV infection. The lack of detections in the foetal tissues of HeV-positive females suggests that vertical transmission is not a regular mode of transmission in naturally infected flying-foxes, and that placental and foetal tissues are not a major source of infection for horses. A better understanding of HeV tissue tropism will strengthen management of the risk of spillover from flying-foxes to horses and ultimately humans.

Temporal Variation in Physiological Biomarkers in Black Flying-Foxes (Pteropus alecto), Australia

Bats of the genus Pteropus (Pteropodidae) are recognised as the natural host of multiple emerging pathogenic viruses of animal and human health significance, including henipaviruses, lyssaviruses and ebolaviruses. Some studies have suggested that physiological and ecological factors may be associated with Hendra virus infection in flying-foxes in Australia; however, it is essential to understand the normal range and seasonal variability of physiological biomarkers before seeking physiological associations with infection status. We aimed to measure a suite of physiological biomarkers in P. alecto over time to identify any seasonal fluctuations and to examine possible associations with life-cycle and environmental stressors. We sampled 839 adult P. alecto in the Australian state of Queensland over a 12-month period. The adjusted population means of every assessed hematologic and biochemical parameter were within the reported reference range on every sampling occasion. However, within this range, we identified significant temporal variation in these parameters, in urinary parameters and body condition, which primarily reflected the normal annual life cycle. We found no evident effect of remarkable physiological demands or nutritional stress, and no indication of clinical disease driving any parameter values outside the normal species reference range. Our findings identify underlying temporal physiological changes at the population level that inform epidemiological studies and assessment of putative physiological risk factors driving Hendra virus infection in P. alecto. More broadly, the findings add to the knowledge of Pteropus populations in terms of their relative resistance and resilience to emerging infectious disease.

Diagnostic values for skin temperature assessment to detect diabetes-related foot complications

Background Skin temperature assessment is a promising modality for early detection of diabetic foot problems, but its diagnostic value has not been studied. Our aims were to investigate the diagnostic value of different cutoff skin temperature values for detecting diabetes-related foot complications such as ulceration, infection, and Charcot foot and to determine urgency of treatment in case of diagnosed infection or a red-hot swollen foot. Materials and Methods The plantar foot surfaces of 54 patients with diabetes visiting the outpatient foot clinic were imaged with an infrared camera. Nine patients had complications requiring immediate treatment, 25 patients had complications requiring non-immediate treatment, and 20 patients had no complications requiring treatment. Average pixel temperature was calculated for six predefined spots and for the whole foot. We calculated the area under the receiver operating characteristic curve for different cutoff skin temperature values using clinical assessment as reference and defined the sensitivity and specificity for the most optimal cutoff temperature value. Mean temperature difference between feet was analyzed using the Kruskal–Wallis tests. Results The most optimal cutoff skin temperature value for detection of diabetes-related foot complications was a 2.2°C difference between contralateral spots (sensitivity, 76%; specificity, 40%). The most optimal cutoff skin temperature value for determining urgency of treatment was a 1.35°C difference between the mean temperature of the left and right foot (sensitivity, 89%; specificity, 78%). Conclusions Detection of diabetes-related foot complications based on local skin temperature assessment is hindered by low diagnostic values. Mean temperature difference between two feet may be an adequate marker for determining urgency of treatment.

Inflammation-driven bone formation in a mouse model of ankylosing spondylitis: Sequential not parallel processes

Background Ankylosing spondylitis (AS) is an immune-mediated arthritis particularly targeting the spine and pelvis and is characterised by inflammation, osteoproliferation and frequently ankylosis. Current treatments that predominately target inflammatory pathways have disappointing efficacy in slowing disease progression. Thus, a better understanding of the causal association and pathological progression from inflammation to bone formation, particularly whether inflammation directly initiates osteoproliferation, is required. Methods The proteoglycan-induced spondylitis (PGISp) mouse model of AS was used to histopathologically map the progressive axial disease events, assess molecular changes during disease progression and define disease progression using unbiased clustering of semi-quantitative histology. PGISp mice were followed over a 24-week time course. Spinal disease was assessed using a novel semi-quantitative histological scoring system that independently evaluated the breadth of pathological features associated with PGISp axial disease, including inflammation, joint destruction and excessive tissue formation (osteoproliferation). Matrix components were identified using immunohistochemistry. Results Disease initiated with inflammation at the periphery of the intervertebral disc (IVD) adjacent to the longitudinal ligament, reminiscent of enthesitis, and was associated with upregulated tumor necrosis factor and metalloproteinases. After a lag phase, established inflammation was temporospatially associated with destruction of IVDs, cartilage and bone. At later time points, advanced disease was characterised by substantially reduced inflammation, excessive tissue formation and ectopic chondrocyte expansion. These distinct features differentiated affected mice into early, intermediate and advanced disease stages. Excessive tissue formation was observed in vertebral joints only if the IVD was destroyed as a consequence of the early inflammation. Ectopic excessive tissue was predominantly chondroidal with chondrocyte-like cells embedded within collagen type II- and X-rich matrix. This corresponded with upregulation of mRNA for cartilage markers Col2a1, sox9 and Comp. Osteophytes, though infrequent, were more prevalent in later disease. Conclusions The inflammation-driven IVD destruction was shown to be a prerequisite for axial disease progression to osteoproliferation in the PGISp mouse. Osteoproliferation led to vertebral body deformity and fusion but was never seen concurrent with persistent inflammation, suggesting a sequential process. The findings support that early intervention with anti-inflammatory therapies will be needed to limit destructive processes and consequently prevent progression of AS.

Expected Performance of the ATLAS Experiment - Detector, Trigger and Physics

A detailed study is presented of the expected performance of the ATLAS detector. The reconstruction of tracks, leptons, photons, missing energy and jets is investigated, together with the performance of b-tagging and the trigger. The physics potential for a variety of interesting physics processes, within the Standard Model and beyond, is examined. The study comprises a series of notes based on simulations of the detector and physics processes, with particular emphasis given to the data expected from the first years of operation of the LHC at CERN.

Designed Cyclic Permutants of HIV-1 gp120: Implications for Envelope Trimer Structure and Immunogen Design

Most HIV-1 broadly neutralizing antibodies are directed against the gp120 subunit of the env surface protein. Native env consists of a trimer of gp120-gp41 heterodimers, and in contrast to monomeric gp120, preferentially binds CD4 binding site (CD4bs)-directed neutralizing antibodies over non-neutralizing ones. Some cryo-electron tomography studies have suggested that the V1V2 loop regions of gp120 are located close to the trimer interface. We have therefore designed cyclically permuted variants of gp120 with and without the h-CMP and SUMO2a trimerization domains inserted into the V1V2 loop. h-CMP-V1cyc is one such variant in which residues 153 and 142 are the N- and C-terminal residues, respectively, of cyclically permuted gp120 and h-CMP is fused to the N-terminus. This molecule forms a trimer under native conditions and binds CD4 and the neutralizing CD4bs antibodies b12 with significantly higher affinity than wild-type gp120. It binds non-neutralizing CD4bs antibody F105 with lower affinity than gp120. A similar derivative, h-CMP-V1cycl, bound the V1V2 loop-directed broadly neutralizing antibodies PG9 and PG16 with similar to 20-fold higher affinity than wild-type JRCSF gp120. These cyclic permutants of gp120 are properly folded and are potential immunogens. The data also support env models in which the V1V2 loops are proximal to the trimer interface.

Polymorphs, Salts, and Cocrystals: What's in a Name?

The December 2011 release of a draft United States Food and Drug Administration (FDA) guidance concerning regulatory classification of pharmaceutical cocrystals of active pharmaceutical ingredients (APIs) addressed two matters of topical interest to the crystal engineering and pharmaceutical science communities: (1) a proposed definition of cocrystals; (2) a proposed classification of pharmaceutical cocrystals as dissociable ``API-excipient'' molecular complexes. The Indo U.S. Bilateral Meeting sponsored by the Indo-U.S. Science and Technology Forum titled The Evolving Role of Solid State Chemistry in Pharmaceutical Science was held in Manesar near Delhi, India, from February 2-4, 2012. A session of the meeting was devoted to discussion of the FDA guidance draft. The debate generated strong consensus on the need to define cocrystals more broadly and to classify them like salts. It was also concluded that the diversity of API crystal forms makes it difficult to classify solid forms into three categories that are mutually exclusive. This perspective summarizes the discussion in the Indo-U.S. Bilateral Meeting and includes contributions from researchers who were not participants in the meeting.

Skeletal Muscle Degeneration and Regeneration in Mice and Flies

Many aspects of skeletal muscle biology are remarkably similar between mammals and tiny insects, and experimental models of mice and flies (Drosophila) provide powerful tools to understand factors controlling the growth, maintenance, degeneration (atrophy and necrosis), and regeneration of normal and diseased muscles, with potential applications to the human condition. This review compares the limb muscles of mice and the indirect flight muscles of flies, with respect to the mechanisms of adult myofiber formation, homeostasis, atrophy, hypertrophy, and the response to muscle degeneration, with some comment on myogenic precursor cells and common gene regulatory pathways. There is a striking similarity between the species for events related to muscle atrophy and hypertrophy, without contribution of any myoblast fusion. Since the flight muscles of adult flies lack a population of reserve myogenic cells (equivalent to satellite cells), this indicates that such cells are not required for maintenance of normal muscle function. However, since satellite cells are essential in postnatal mammals for myogenesis and regeneration in response to myofiber necrosis, the extent to which such regeneration might be possible in flight muscles of adult flies remains unclear. Common cellular and molecular pathways for both species are outlined related to neuromuscular disorders and to age-related loss of skeletal muscle mass and function (sarcopenia). The commonality of events related to skeletal muscles in these disparate species (with vast differences in size, growth duration, longevity, and muscle activities) emphasizes the combined value and power of these experimental animal models.

Skeletal Muscle Degeneration and Regeneration in Mice and Flies

Many aspects of skeletal muscle biology are remarkably similar between mammals and tiny insects, and experimental models of mice and flies (Drosophila) provide powerful tools to understand factors controlling the growth, maintenance, degeneration (atrophy and necrosis), and regeneration of normal and diseased muscles, with potential applications to the human condition. This review compares the limb muscles of mice and the indirect flight muscles of flies, with respect to the mechanisms of adult myofiber formation, homeostasis, atrophy, hypertrophy, and the response to muscle degeneration, with some comment on myogenic precursor cells and common gene regulatory pathways. There is a striking similarity between the species for events related to muscle atrophy and hypertrophy, without contribution of any myoblast fusion. Since the flight muscles of adult flies lack a population of reserve myogenic cells (equivalent to satellite cells), this indicates that such cells are not required for maintenance of normal muscle function. However, since satellite cells are essential in postnatal mammals for myogenesis and regeneration in response to myofiber necrosis, the extent to which such regeneration might be possible in flight muscles of adult flies remains unclear. Common cellular and molecular pathways for both species are outlined related to neuromuscular disorders and to age-related loss of skeletal muscle mass and function (sarcopenia). The commonality of events related to skeletal muscles in these disparate species (with vast differences in size, growth duration, longevity, and muscle activities) emphasizes the combined value and power of these experimental animal models.

Estudio de la efectividad garrapaticida del compuesto Cypergan-15 en las razas Aberdeen Angus y Brahman en el departamento de Boaco, Nicaragua

El presente trabajo se realizó en la UPE Santa Fe de la Empresa Genética Agenor Gómez, localizada a 15 Km al Noreste de la ciudad de Boaco, Nicaragua. Evaluándose la efectividad del garrapaticida Cipergan-15 mediante el recuento del número de garrapatas vivas después de la aplicación de diferentes dosis (0.0, 0.5, 1.0 y 1.5 ml/lt de agua) en prueba In Vitro e In Vivo. En la primera, a periodos de recuentos de 3, 18, 24 y 42 hrs y en la última, a periodos de 1, 2, 3, 7 y 14 días después de la aplicación en las razas Angus y Brahrnan. Los resultados obtenidos en la prueba In Vitro demostraron que el producto posee efecto garrapaticida, alcanzando un 100% de control a concentraciones de 1.5 y 1.0 ml/lt de agua, al cabo de 3 y 24 hrs respectivamente. En el análisis de varianza para la prueba In Vivo, los resultad indican que existen diferencias significativas en el número de garrapatas vivas en cada una de las concentraciones estudiadas, exceptuando la comparación al séptimo dia de recuento entre los niveles de 1.0 ml/lt de agua y 1.5 ml/lt de agua. El efecto de raza. También resultó significativo en el 14avo día y la interacción raza-concentración a los 1, 2 y 14 días de recuentos. Al efectuarse la prueba de "t" para medias ajustadas se encontró que las concentraciones de 1.0 ml/lt de agua y 1.5 m1/lt de agua ejercen el mayor control en todo el periodo del ensayo. La concentración de 1.5 rnl/lt de agua, disminuye el número de garrapatas a cero a los 3 días post­ aplicación, manteniéndolas aún en el séptimo día y con promedios inferiores de 8 garrapatas en el 14avo día. La concentración de 1.0 ml/lt de agua disminuye el número de garrapatas a promedios inferiores de 2 a los 3 días, menores que 1 al 7mo día e inferiores a 16 a los 14 días en ambas razas. En cambio, en el grupo testigo, el número de garrapatas se incrementó a una tasa promedio de 37%. Los estimados de la concentración minina para mantener una población promedio de garrapatas por debajo de 20 aún en el 14avo día, resultó de 0.86 ml/lt de agua para la raza Brahman y de 0.96 ml/lt de agua para la raza Angus.

Influencia de humedad y temperatura sobre la fertilidad de vacas holstein, en la Península de Chiltepe

Se analizó los resultados de 2,419 inseminaciones al primer servicio (782 en vaquillas y 1,637 en vacas) realizadas en la Empresa Genética Roberto Alvarado (EGRA), ubicada en la península de Chiltepe (km 24 carretera a León), durante los años 1987 y 1988. Estos resultados fueron cotejados con los datos de humedad y temperatura ambiente del día de su realización provenientes de la estación meteorológica Agusto Cesar Sandino, ubicada en el Aeropuerto Internacional de Managua. Se estudió la influencia de cuatro variables de temperatura, lo mismo que de humedad relativa: Máxima diaria, mínima diaria, promedio diario y promedio diurno. Además se estudió la combinación de temperatura y humedad: Máxima diaria con máxima diaria, mínima diaria con mínima diaria, etc. Se encontró que la fertilidad de las vaquillas parece no ser influenciada por ninguna de las variables estudiadas, a excepción de HMX Humedad Relativa Máxima del día de inseminación) cuya influencia parece ser muy débil. De acuerdo con las Pruebas de Independencia de Chi cuadrado las variables más relacionadas con la fertilidad de las vacas son HDA (Humedad Relativa Promedio del día de inseminación) y HMX. Las variables de temperatura ambiental más relacionadas con la fertilidad de las vacas son TDA (Temperatura Promedio del día de inseminación) y TDU (Temperatura Promedio diurna del día de inseminación). Un incremento de TDA o TDU parece afectar positivamente la fertilidad de las vacas con valores de TDA 24 – 29°c se obtuvieron tasas de concepción entre 34 y 35% y con valores de TDA de 29- 31.5°c se obtuvo una tasa de concepción del 47%. Todas las variables de humedad relativa parecen afectar negativamente la fertilidad de las vacas: la tasa de concepción bajó de un 46.6% con valores de HDA de 48 – 65% a un 29.7% con valores de HDA de 80 – 96%. Al estudiar el efecto combinada de temperatura y humedad se muestra una tendencia de la tasas de concepción a disminuir conforme la humedad relativa aumenta y a incrementarse 1igeramente conforme aumenta 1a temperatura, aun cuando esta última tendencia puede variar según el nivel de humedad presente. Los valores medios de temperatura combinados con valores medios y altos de humedad resultan muy adversos a la fertilidad de las vacas. La Humedad Relativa parece ser el principal factor responsable de la caída en las tasas de concepción durante algunos meses del año.

Evaluación de cinco cepas de Rhizobium leguminosarum bv. phaseoli en el cultivo del frijol (Phaseolus vulgaris L.) var. Rev. 84

Esta investigación se realizó en el Centro Experimental "La Compañía" ubicada en San Marcos, Carazo, con el objetivo de evaluar la adaptabilidad de las cepas en las condiciones estudiadas, determinar cuál de las cepas es la más infectiva y efectiva en cuanto a la nodulación y determinar su efecto en el crecimiento y rendimiento del cultivo del frijol (Phaseolus vulgaris L.) var. Rev. 84. Los factores evaluados fueren 5 cepas de Rhisoblum leguminosearum by. phaseoli y 3 testigos: 1 Testigo Absoluto (no inoculado. no fertilizado) Y 2 Testigos Relativos (no inoculados y fertilizados). El diseño experimental utilizado fue el de bloques completos al azar (SCA), con 4 repeticiones. La parcela experimental del ensayo fue de 32 (8 m x 4 m), la distancia de siembra fue de 0.1 m entre plantas y 0.4 m entre surcos. Las variables analizadas fueron: número; eficiencia y peso seco de nódulos por planta, peso seco de área foliar y rendimiento. Los datos se procesaron usando análisis de varianza (AHUEVA) y se utilizó la prueba de rangos múltiples de DUNCAN (P ≤ 0. 05). No se observó diferencias significativas para las variables de nodulación y rendimiento, determinándose efecto significativo sólo para el peso seco de área foliar en la I y II etapa de evaluación Sin embargo, se determinó quo la cepa CR-436 fue la que presentó mayor infectividad en el sistema radicular y la cepa CIAT-899, la que presentó el mejor porcentaje de efectividad nodular y con la que se obtuvo el mayor promedio de peso seco de área foliar. El mejor rendimiento fue obtenido con la cepa CR-436.

Estudio de tres líneas promisorias de arroz (Oryza sativa L.) en comparación con cinco variedades comerciales en condiciones de secano medianamente favorecido

El presente estudio se realizó en fincas de productores privados en tres localidades del departamento de Chinandega. El propósito fue evaluar tres líneas promisorias de Arroz en comparación con cinco variedades comerciales en condiciones de secano medianamente favorecido. El diseño experimental utilizado fue el de bloques completos al azar con tres repeticiones, utilizando una distancia entre surcos de 30 cm con siembra a chorrillo. En base a los resultados obtenidos se llegó a las conclusiones siguientes: el mejor material con respecto a características agronómicas, componentes de rendimiento y rendimiento agrícola fue Altamira-9; la línea CT-5754 es un material muy promisorio debido principalmente a su precocidad, característica muy importante para las zonas en que las precipitaciones son muy erráticas; En Ameya una de las localidades evaluadas, todos los materiales mostraron un mejor comportamiento.