Molecular surveillance detects high prevalence of the neglected parasite Mansonella ozzardi in the Colombian Amazon.

Mansonellosis is an undermapped insect-transmitted disease caused by filarial nematodes that are estimated to infect hundreds of millions of people globally. Despite their prevalence, there are many outstanding questions regarding the general biology and health impacts of the responsible parasites. Historical reports suggest that the Colombian Amazon is endemic for mansonellosis and may serve as an ideal location to pursue these questions in the backdrop of other endemic and emerging pathogens. We deployed molecular and classical diagnostic approaches to survey Mansonella prevalence among adults belonging to indigenous communities along the Amazon River and its tributaries near Leticia, Colombia. Deployment of a loop-mediated isothermal amplification (LAMP) assay on blood samples revealed an infection prevalence of ∼40% for Mansonella ozzardi . This assay identified significantly more infections than blood smear microscopy or LAMP assays performed using plasma, likely reflecting greater sensitivity and the ability to detect low microfilaremias or occult infections. Mansonella infection rates increased with age and were higher among males compared to females. Genomic analysis confirmed the presence of M. ozzardi that clusters closely with strains sequenced in neighboring countries. We successfully cryopreserved and revitalized M. ozzardi microfilariae, advancing the prospects of rearing infective larvae in controlled settings. These data suggest an underestimation of true mansonellosis prevalence, and we expect that these methods will help facilitate the study of mansonellosis in endemic and laboratory settings.

Optical flow driven interpolation for isotropic FIB-SEM reconstructions.

BACKGROUND AND OBJECTIVE: Focused Ion Beam - Scanning Electron Microscopy (FIB-SEM) allows three-dimensional ultrastructural analysis of cells and tissues at the nanoscale. The technique iteratively removes a section of the sample with a FIB and takes an SEM image from the exposed surface. The section thickness is usually higher than the image pixel size to reduce acquisition time, thus resulting in anisotropic resolution. In this work, we explore novel interpolation methods along the sectioning direction to produce isotropic resolution and facilitate proper interpretation of the FIB-SEM 3D volumes. METHODS: Classical interpolation methods are usually applied in this context under the assumption that the changes through successive images are relatively smooth. However, the actual 3D arrangement of the structures in the sample may cause significant changes in the biological features between consecutive images of the FIB-SEM stacks. We have developed a novel interpolation strategy that accounts for this variation by using the Optical Flow (OF) to estimate it. As an intermediate stage, OF-compensated images are produced by aligning the spatial regions of the biological structures. Interpolated images are then generated from these OF-compensated images. The final isotropic stack is assembled by interleaving the interpolated images with the original images of the anisotropic stack. RESULTS: OF-driven and classical interpolation methods were compared using an objective assessment based on Pearson Correlation Coefficient (PCC) and a qualitative evaluation based on visual results, using public datasets and representative anisotropy conditions. The objective assessment demonstrated that the OF-driven interpolation always yields higher PCC values, with interpolated images closer to the ground truth. The qualitative evaluation corroborated those results and confirmed that classical interpolation may blur areas with substantial changes between consecutive images whereas OF-driven interpolation provides sharpness. CONCLUSIONS: We have developed an OF-driven interpolation approach to generating FIB-SEM stacks with isotropic resolution from experimental anisotropic data. It adapts to the rapid variation of the biological structures observed through the images of the FIB-SEM stack. Our approach outperforms classical interpolation and manages to produce sharp interpolated views in cases where there are significant changes between consecutive experimental images.

Temporal and spatial expression of genes involved in DNA methylation during reproductive development of sexual and apomictic Eragrostis curvula.

Recent reports in model plant species have highlighted a role for DNA methylation pathways in the regulation of the somatic-to-reproductive transition in the ovule, suggesting that apomixis (asexual reproduction through seeds) likely relies on RdDM downregulation. Our aim was therefore to explore this hypothesis by characterizing genes involved in DNA methylation in the apomictic grass Eragrostis curvula. We explored floral transcriptomes to identify homologs of three candidate genes, for which mutations in Arabidopsis and maize mimic apomixis (AtAGO9/ZmAGO104, AtCMT3/ZmDMT102/ZmDMT105, and AtDDM1/ZmCHR106), and compared both their spatial and temporal expression patterns during reproduction in sexual and apomictic genotypes. Quantitative expression analyses revealed contrasting expression patterns for the three genes in apomictic vs sexual plants. In situ hybridization corroborated these results for two candidates, EcAGO104 and EcDMT102, and revealed an unexpected ectopic pattern for the AGO gene during germ line differentiation in apomicts. Although our data partially support previous results obtained in sexual plant models, they suggest that rather than an RdDM breakdown in the ovule, altered localization of AtAGO9/ZmAGO104 expression is required for achieving diplospory in E. curvula. The differences in the RdDM machinery acquired during plant evolution might have promoted the emergence of the numerous apomictic paths observed in plants.

Sharp Morphological Transitions from Nanoscale Mixed-Anchoring Patterns in Confined Nematic Liquid Crystals.

Liquid crystals are known to be particularly sensitive to orientational cues provided at surfaces or interfaces. In this work, we explore theoretically, computationally, and experimentally the behavior of liquid crystals on isolated nanoscale patterns with controlled anchoring characteristics at small length scales. The orientation of the liquid crystal is controlled through the use of chemically patterned polymer brushes that are tethered to a surface. This system can be engineered with remarkable precision, and the central question addressed here is whether a characteristic length scale exists at which information encoded on a surface is no longer registered by a liquid crystal. To do so, we adopt a tensorial description of the free energy of the hybrid liquid-crystal-surface system, and we investigate its morphology in a systematic manner. For long and narrow surface stripes, it is found that the liquid crystal follows the instructions provided by the pattern down to 100 nm widths. This is accomplished through the creation of line defects that travel along the sides of the stripes. We show that a "sharp" morphological transition occurs from a uniform undistorted alignment to a dual uniform/splay-bend morphology. The theoretical and numerical predictions advanced here are confirmed by experimental observations. Our combined analysis suggests that nanoscale patterns can be used to manipulate the orientation of liquid crystals at a fraction of the energetic cost that is involved in traditional liquid crystal-based devices. The insights presented in this work have the potential to provide a new fabrication platform to assemble low power bistable devices, which could be reconfigured upon application of small external fields.

Effects of the Ottonia martiana Miq. (Piperaceae) extract on dog's ocular surface / Efeitos do extrato de Ottonia martiana Miq. (Piperaceae) sobre a superfície ocular de cães

The anesthetics effects of aqueous extract of Ottonia martiana leaves were studied on the ocular surface of healthy beagle dogs. The dogs were divided in three groups (n=15): control group (CG), proxymetacaine group (PG) and Ottonia group (OG), which were treated with 0.9% saline, 0.5% proxymetacaine hydrochloride ophthalmic solution and O. martiana extract respectively. An oftalmic evaluation was performed before the treatments. Eye drops were instilled at time 0 (T0) and 3 minutes later (T3). Axial corneal sensitivity was evaluated by esthesiometry 5 and 10 minutes after T0. Tear production and intraocular pressure were evaluated 10 minutes after T0. Slit lamp biomicroscopy was performed 10 and 20 minutes after T0 and the eyes were stained with fluorescein 20 minutes after T0. The STT was reduced in PG. Conjunctival hyperemia was observed in 13 animals from PG and constituted the only ocular alteration observed during the study. Esthesiometry revealed a decreased corneal sensitivity for PG and OG. Those results show that the O. martiana extract acts reducing corneal sensitivity in dogs. Moreover, its use does not decrease the tear production and does not cause any clinical ophthalmic alteration.

Estudaram-se os efeitos do extrato das folhas de Ottonia martiana sobre a superfície ocular de cães hígidos da raça Beagle. Compuseram-se três grupos de tratamento (n=15): grupo controle (GC), grupo proximetacaína (GP) e grupo Ottonia (GO), tratados, respectivamente, com solução fisiológica, colírio de cloridrato de proximetacaína a 0,5% e extrato de O. martiana. Após avaliação oftálmica inicial, os tratamentos foram realizados no tempo 0 (T0) e decorridos 3min (T3). Avaliaram-se a sensibilidade axial da córnea por estesiometria (T5 e T10) e a produção lacrimal e a pressão ocular (T10). Realizaram-se a biomicroscopia com lâmpada em fenda (T10 e T20), e o teste do tingimento pela fluoresceína (T20). Relativamente ao teste de Schirmer, observou-se diminuição nos cães do GP. Houve alteração clínica somente nos do GP, em que 13 animais apresentaram hiperemia conjuntival. Relativamente à estesiometria, houve diminuição da sensibilidade corneal nos animais do GP e do GO. Admite-se que o extrato de O. martiana age diminuindo a sensibilidade corneal em cães e que sua utilização não diminui a produção lacrimal, tampouco causa alterações clínicas oftálmicas.

Liquid-crystal-mediated self-assembly at nanodroplet interfaces.

Technological applications of liquid crystals have generally relied on control of molecular orientation at a surface or an interface. Such control has been achieved through topography, chemistry and the adsorption of monolayers or surfactants. The role of the substrate or interface has been to impart order over visible length scales and to confine the liquid crystal in a device. Here, we report results from a computational study of a liquid-crystal-based system in which the opposite is true: the liquid crystal is used to impart order on the interfacial arrangement of a surfactant. Recent experiments on macroscopic interfaces have hinted that an interfacial coupling between bulk liquid crystal and surfactant can lead to a two-dimensional phase separation of the surfactant at the interface, but have not had the resolution to measure the structure of the resulting phases. To enhance that coupling, we consider the limit of nanodroplets, the interfaces of which are decorated with surfactant molecules that promote local perpendicular orientation of mesogens within the droplet. In the absence of surfactant, mesogens at the interface are all parallel to that interface. As the droplet is cooled, the mesogens undergo a transition from a disordered (isotropic) to an ordered (nematic or smectic) liquid-crystal phase. As this happens, mesogens within the droplet cause a transition of the surfactant at the interface, which forms new ordered nanophases with morphologies dependent on surfactant concentration. Such nanophases are reminiscent of those encountered in block copolymers, and include circular, striped and worm-like patterns.

Liquid crystal mediated interactions between nanoparticles in a nematic phase.

A continuum theory is used to study the interactions between nanoparticles suspended in nematic liquid crystals. The free energy functional that describes the system is minimized using an Euler-Lagrange approach and an unsymmetric radial basis function method. It is shown that nanoparticle liquid-crystal mediated interactions can be controlled over a large range of magnitudes through changes of the anchoring energy and the particle diameter. The results presented in this work serve to reconcile past discrepancies between theoretical predictions and experimental observations, and suggest intriguing possibilities for directed nanoparticle self-assembly in liquid crystalline media.

Ultrasonographic analysis of senile cataractous lens of dogs and its correlation to phacoemulsification / Análise ultrassonográfica da lente cataratogênica senil de cães e sua correlação com parâmetros de facoemulsificação

This study aimed to evaluate the effectiveness of a computer-assisted ultrasound analysis of senile dogs cataractous lens and identify its correlation to phacoemulsification surgical parameters. Simultaneous A- and B- mode ultrasonic evaluation of the lens was performed on 15 dogs with cataract. A computer-assisted image analysis was used to objectively calculate the lens echogenicity. Following ultrasonographic examination the cataracts were removed by phacoemulsification and the ultrasonographic and phacoemulsification findings were compared to identify possible correlations. Lens echogenicity obtained by the computer-assisted ultrasonographic analysis correlated to the phacoemulsification time, such that the whiter the ultrasound image, the longer was the phacoemulsification time. The B-mode ultrasonography was helpful for the prediction of lens opacity location, but not predictive of lens hardness.

Avaliou-se a eficácia da análise ultrassonográfica, assistida por computador, da lente cataratogênica senil de cães e identificou-se sua correlação com parâmetros da facoemulsificação. A avaliação ultrassonográfica da lente em modos A e B simultâneos foi realizada em 15 cães portadores de catarata. Procedeu-se à análise das imagens, assistida por computador. Posteriormente, os olhos selecionados foram submetidos à facoemulsificação. Os achados ultrassonográficos e a facoemulsificação foram comparados visando à identificação de possíveis correlações. A ecogenicidade obtida por meio da análise ultrassonográfica correlacionou-se diretamente com o tempo de facoemulsificação, de maneira que quanto mais branca a imagem ultrassonográfica, maior o tempo de facoemulsificação. A ultrassonografia em modo-B auxiliou na predição da localização das opacidades lenticulares, porém não auxiliou na predição da dureza da lente.

Drag reduction by polyethylene glycol in the tail arterial bed of normotensive and hypertensive rats

This study was designed to evaluate the effect of drag reducer polymers (DRP) on arteries from normotensive (Wistar) and spontaneously hypertensive rats (SHR). Polyethylene glycol (PEG 4000 at 5000 ppm) was perfused in the tail arterial bed with (E+) and without endothelium (E-) from male, adult Wistar (N = 14) and SHR (N = 13) animals under basal conditions (constant flow at 2.5 mL/min). In these preparations, flow-pressure curves (1.5 to 10 mL/min) were constructed before and 1 h after PEG 4000 perfusion. Afterwards, the tail arterial bed was fixed and the internal diameters of the arteries were then measured by microscopy and drag reduction was assessed based on the values of wall shear stress (WSS) by computational simulation. In Wistar and SHR groups, perfusion of PEG 4000 significantly reduced pulsatile pressure (Wistar/E+: 17.5 ± 2.8; SHR/E+: 16.3 ± 2.7 percent), WSS (Wistar/E+: 36; SHR/E+: 40 percent) and the flow-pressure response. The E- reduced the effects of PEG 4000 on arteries from both groups, suggesting that endothelial damage decreased the effect of PEG 4000 as a DRP. Moreover, the effects of PEG 4000 were more pronounced in the tail arterial bed from SHR compared to Wistar rats. In conclusion, these data demonstrated for the first time that PEG 4000 was more effective in reducing the pressure-flow response as well as WSS in the tail arterial bed of hypertensive than of normotensive rats and these effects were amplified by, but not dependent on, endothelial integrity. Thus, these results show an additional mechanism of action of this polymer besides its mechanical effect through the release and/or bioavailability of endothelial factors.

Drag reduction by polyethylene glycol in the tail arterial bed of normotensive and hypertensive rats.

This study was designed to evaluate the effect of drag reducer polymers (DRP) on arteries from normotensive (Wistar) and spontaneously hypertensive rats (SHR). Polyethylene glycol (PEG 4000 at 5000 ppm) was perfused in the tail arterial bed with (E+) and without endothelium (E-) from male, adult Wistar (N = 14) and SHR (N = 13) animals under basal conditions (constant flow at 2.5 mL/min). In these preparations, flow-pressure curves (1.5 to 10 mL/min) were constructed before and 1 h after PEG 4000 perfusion. Afterwards, the tail arterial bed was fixed and the internal diameters of the arteries were then measured by microscopy and drag reduction was assessed based on the values of wall shear stress (WSS) by computational simulation. In Wistar and SHR groups, perfusion of PEG 4000 significantly reduced pulsatile pressure (Wistar/E+: 17.5 ± 2.8; SHR/E+: 16.3 ± 2.7%), WSS (Wistar/E+: 36; SHR/E+: 40%) and the flow-pressure response. The E- reduced the effects of PEG 4000 on arteries from both groups, suggesting that endothelial damage decreased the effect of PEG 4000 as a DRP. Moreover, the effects of PEG 4000 were more pronounced in the tail arterial bed from SHR compared to Wistar rats. In conclusion, these data demonstrated for the first time that PEG 4000 was more effective in reducing the pressure-flow response as well as WSS in the tail arterial bed of hypertensive than of normotensive rats and these effects were amplified by, but not dependent on, endothelial integrity. Thus, these results show an additional mechanism of action of this polymer besides its mechanical effect through the release and/or bioavailability of endothelial factors.

Recobrimento conjuntival em 360º e ceratectomia superficial na ceratite bolhosa em cão: relato de caso / Superficial keratectomy and 360º conjunctival flap for bullous keratopathy in a dog: a case report

A case of a two-year-old male Pinscher with a history of discomfort in the right eye was reported. The left eye had been enucleated by the referring veterinarian due to the same symptom with unsuccessful clinical treatment. The Schirmer tear test value was elevated and a decreased intraocular pressure was observed by applanation tonometry. Biomicroscopy revealed profuse corneal edema and keratoconus and fluorescein staining was negative. Gonioscopy and ophthalmoscopy did not provide any relevant data due to the corneal alterations. Bullous keratopathy was diagnosed. Surgery was performed in two steps: 1) superficial keratectomy and 360º conjunctival flap, and 2) superficial keratectomy to restore corneal transparency. Thirty days after the second superficial keratectomy, the third eyelid flap was removed. Conjunctivalization of the upper nasal quadrant of the cornea was observed. The axial portion of the cornea was transparent and vision was restored.

Relata-se o caso ocorrido em um cão, da raça Pinscher, com dois anos de idade e histórico de desconforto no olho direito. O olho esquerdo havia sido enucleado por outro profissional, por apresentar os mesmos sinais, cujo tratamento clínico instituído não lograra êxito. O valor do teste da lágrima de Schirmer encontrava-se aumentado e identificou-se diminuição da pressão intraocular à tonometria de aplanação. Observaram-se, à biomicroscopia, edema corneal profuso e ceratocone, e o teste da fluoresceína foi negativo. Gonioscopia e oftalmoscopia não lograram fornecer dados relevantes dadas as condições da córnea. Diagnosticou-se ceratite bolhosa. Optou-se pelo tratamento cirúrgico, que fora realizado em duas etapas: 1- ceratectomia superficial e "flap" conjuntival de 360º; 2- ceratectomia superficial para devolver transparência à córnea. Transcorridos 30 dias da segunda ceratectomia superficial, o "flap" de terceira pálpebra foi desfeito. Observou-se conjuntivalização do quadrante nasal superior da córnea, córnea clara no eixo visual e retorno da visão.

Hemangiosarcoma of the nictitant membrane in a Brazilian Fila dog: case report / Hemangiosarcoma em membrana nictitante de cão da ração Fila Brasileiro: relato de caso

It was reported a case of a 9-year-old Brazilian Fila male dog that had been affected by a tissular formation of the nictitant membrane of the right eye with four months of evolution. Conjunctival hyperemia and edema, serosanguinolent secretion, and a conjunctival neoformation at the nictitant membrane were observed. Surgical exeresis of the neoformation was performed. Endothelial cells slightly differentiated and pleomorphic, with an intense vascular neoformation compatible to nictitant membrane hemangiosarcoma, were found at the histological exam.

Relata-se o caso de um cão, macho, da raça Fila Brasileiro, com nove anos de idade, acometido por neoformação tecidual na membrana nictitante do olho direito, com cerca de quatro meses de evolução. Realizou-se exame oftálmico rotineiro, a partir do qual se notaram hiperemia e edema conjuntivais, secreção ocular serossangüinolenta e neoformação saliente na conjuntiva da membrana nictitante. Realizou-se a exérese cirúrgica da neoformação. À histopatologia, encontraram-se células endoteliais pouco diferenciadas e pleomórficas que originavam intensa neoformação vascular, compatíveis com hemangiossarcoma da membrana nictitante.

Intraocular pressure and its correlation with blood gas parameters in crested caracaras (Caracara plancus) anesthetized with isoflurane and sevoflurane / Pressão intra-ocular e sua correlação com parâmetros hemogasométricos em caracarás (Caracara plancus) anestesiados com isofluorano e sevofluorano

The intraocular pressure (IOP) and its correlations with arterial carbon dioxide partial pressure (PaCO2) and arterial pH were studied in five crested caracaras (Caracara plancus) anesthetized with isoflurane (ISO) and sevoflurane (SEV). Baseline IOP values were measured in both eyes (M0). Brachial artery was previously catheterized to obtain blood gas and cardiorespiratory analysis. Anesthesia was induced with 5 percent ISO and maintained with 2.5 percent for 40 minutes. IOP measurements and blood samples were evaluated in different moments until the end of the procedure. After recovering, a second anesthesia was induced with 6 percent SEV and maintained with 3.5 percent. Parameters were evaluated at the same time points of the previous procedure. IOP reduced significantly (P= 0.012) from M0 at all time points and no significative changes were observed between ISO and SEV anesthesias. Correlation between IOP and PaCO2 and between PIO and blood pH were found only for SEV. IOP and blood pH decreased in parallel with IOP, whereas values of PaCO2 increased in caracaras anesthetized with isoflurane and sevoflurane.

Avaliou-se a pressão intra-ocular (PIO) e estimaram-se as correlações entre PIO e pressão de dióxido de carbono (PaCO2) e pH arterial de cinco caracarás (Caracara plancus), anestesiados com isofluorano (ISO) ou sevofluorano (SEV). Valores basais da PIO foram aferidos em ambos os olhos (M0). Cateterizou-se previamente a artéria braquial para obtenção de parâmetros hemogasométricos e cardiorrespiratórios. Anestesia foi induzida com ISO a 5V por cento e mantida por 40 minutos com 2,5V por cento. PIO e amostras de sangue foram avaliadas em diferentes momentos até o final do procedimento. Após recuperação, uma segunda anestesia foi realizada com SEV a 6 por cento e mantida com 3,5 por cento. Os parâmetros foram aferidos nos mesmos momentos estabelecidos previamente. A PIO decresceu significativamente (P=0,012) de M0 em todos os momentos e não houve diferença estatística entre ISO e SEV. Correlações significativas entre PIO e PaCO2 e entre PIO e pH sangüíneo foram observadas apenas para a anestesia com SEV. O pH sangüíneo decresceu paralelamente a PIO, enquanto a PaCO2 aumentou, em carcarás anestesiados com isofluorano e sevofluorano.

Alpha(2)-noradrenergic antagonist administration into the central nucleus of the amygdala blocks stress-induced hypoalgesia in awake behaving rats.

Stress-induced hypoalgesia (SIH) is an adaptive behavioral phenomenon mediated in part by the amygdala. Acute stress increases amygdalar noradrenaline levels and focal application of alpha(2)-adrenoceptor agonists in the central nucleus of the amygdala (CeA) is antinociceptive. We hypothesized that alpha(2)-adrenoceptor antagonist administration into the CeA may block SIH. Bilateral microinjections of drug or saline via chronically implanted CeA cannulae were followed by either a period of restraint stress or rest. The nocifensive paw-withdrawal latency (PWL) to a focused beam of light was measured. PWLs were longer in restrained rats, constituting SIH. Microinjection of the alpha(2)-adrenoceptor antagonist idazoxan into the CeA prior to restraint blocked SIH. Idazoxan administration in unrestrained rats had no effect. Microinjection of the alpha(2)-adrenoceptor agonist clonidine in unrestrained rats caused dose dependent hypoalgesia, mimicking the effects of environmental stress. alpha(2)-Adrenoceptor function in the CeA is necessary for restraint-induced SIH.

Virtual slide telepathology systems with JPEG2000.

In this paper the JPEG2000 standard is analyzed showing how to exploit all its powerful features in order to build efficient virtual slide telepathology systems. We propose a fast method for stitching images in the wavelet domain. Stitching is a process necessary in many virtual slide systems to generate only one image of a slide with the maximum resolution. This method profits from the structure of the JPEG2000 images to carry out the process with the minimum memory consumption and computational load, obtaining a smoothed union without losing any detail.

Noradrenergic agonist administration into the central nucleus of the amygdala increases the tail-flick latency in lightly anesthetized rats.

The amygdala is a medial forebrain structure with an established role in nociceptive modulation, including the expression of stress-induced hypoalgesia (SIH). Projections from the locus coeruleus increase levels of noradrenaline in the amygdala during acute stress. alpha(2)-Noradrenergic receptor agonists have significant clinical utility as analgesic agents. We therefore hypothesized that alpha(2)-noradrenergic activation of the amygdala may result in behaviorally measurable hypoalgesia. Lightly anesthetized rats underwent microinjection of the alpha(2)-noradrenergic agonist clonidine into the amygdala and intermittent measurement of thermal nociception using the tail-flick latency (TFL). Bilateral microinjection of clonidine into the central nucleus of the amygdala (CeA) resulted in a significant, dose-dependent increase in TFL. This effect was blocked by systemic pre-treatment with the alpha(2)-antagonist yohimbine or by local pre-injection of the alpha(2)-antagonist idazoxan but not by local pre-injection of the alpha(1)-antagonist WB-4101. When injected alone, no antagonist resulted in a significant change in TFL compared with baseline. Clonidine injection into the amygdala but outside the CeA, including the basolateral nucleus of the amygdala, did not significantly alter TFL. These results demonstrate that anatomically and pharmacologically specific activation of alpha(2)-receptors in the CeA in lightly anesthetized rats results in behaviorally measurable antinociception.