The mechanisms that differentiate rabies infections into furious and paralytic forms remain undetermined. There are no neuropathological features in human brains that distinguish furious and paralytic rabies. This could be due to methodology and/or examination of specimens late in the disease course.
In this study, postmortem examination of brain (5 furious and 5 paralytic) and spinal cord (3 furious and 3 paralytic) specimens was performed in 10 rabies-infected dogs, sacrificed shortly after developing the illness. Rabies virus (RABV) antigen (percentage of positive neurons, average antigen area in positive neurons and average antigen area per neuron) and RNA were quantified at 15 different central nervous system (CNS) regions. The distribution and degree of inflammation were also studied.
More RABV antigen was detected in furious rabies than paralytic in many of the CNS regions studied. Caudal-rostral polarity of viral antigen distribution was found in both clinical forms in order from greatest to least: spinal cord, brainstem, cerebellum, midline structures (caudate, thalamus), hippocampus, and cerebrum. In contrast, RABV RNA was most abundant in the cerebral midline structures. Viral RNA was found at significantly higher levels in the cerebral cortex, thalamus, midbrain and medulla of dogs with the furious subtype. The RNA levels in the spinal cord were comparable in both clinical forms. A striking inflammatory response was found in paralytic rabies in the brainstem.
These observations provide preliminary evidence that RABV antigen and RNA levels are higher in the cerebrum in furious rabies compared to the paralytic form. In addition, brainstem inflammation, more pronounced in paralytic rabies, may impede viral propagation towards the cerebral hemispheres.
Rabies is an infectious disease of the central nervous system (CNS) caused by a neurotropic RNA virus in the family Rhabdoviridae, genus Lyssavirus. The worldwide number of rabies deaths estimated by the World Health Organization survey in 1998 was 55,000 annually with the highest incidence in Asia  and this number increased to 70,000 in 2011 . Rabies encephalitis is almost universally fatal, with only six survivors having been recorded to date [4–7]. In Thailand, 10–20 patients (0.03 per 100,000 populations) die of rabies each year .
Rabies can manifest as furious or paralytic forms in humans and dogs. Limbic signs dominate the clinical picture in the former whereas a paralysis of lower motor neuron type dominates the latter . The ratio between furious and paralytic rabies in human cases is approximately 3:1 . Furious rabies patients tend to have a shorter survival (average 5.7 days compared to 11 days in the paralytic form) [9, 11]. The mechanisms that result in two distinct clinical forms remain enigmatic. In human cases, no differences were found in rabies virus (RABV) antigen distribution or the degree of inflammation between furious and paralytic rabies cases at post-mortem . RABV antigen was confined mainly to midline structures (thalamus, brainstem and basal ganglia) in patients who died earlier than 5 days after clinical onset. Several recent clinical, electrophysiological, and neuropathological findings have, however, suggested possible different mechanisms for the furious and paralytic forms of rabies. Peripheral nerve dysfunction, axon- or myelinopathy, is considered to be responsible for the clinical weakness in paralytic rabies . In contrast, in the case of furious rabies, dysfunction of spinal cord anterior horn cells has been identified by electrophysiology, and central chromatolysis by postmortem examination, despite an absence of weakness .
Although previous magnetic resonance imaging (MRI) studies in human rabies patients have revealed no differences between the two clinical forms , diffusion tensor imaging technique has demonstrated disruption of neural tract integrity at the brainstem level in dogs with the paralytic form of rabies . A marked decrease in the mean diffusivity values, representing cytotoxic edema, was noted in the cerebral hemispheres without evidence of blood brain barrier leakage in paralytic dogs . The blood brain barrier was also intact in furious dogs but with minimal cytotoxic brain edema.
Discrepant MRI results between rabies in humans and dogs may be explained by timing of examination, which is late in the course of disease in humans. The current study was, therefore, conducted in rabies-infected dogs, sacrificed shortly after the onset of symptoms. The current study included a neuropathological assessment of the brain and spinal cord, including a determination of the distribution and amount of RABV antigen and inflammatory response. These results were then correlated with antemortem imaging studies. A significantly greater amount of viral antigen was found in many regions of the brain and spinal cord in furious compared to paralytic rabies. This was also true for RABV RNA at regions above the level of the spinal cord. Inflammation was more prominent in the brainstem in paralytic rabies suggesting the inflammatory response may be critical in influencing clinical presentation and disease course.
The selected animals were community dogs with no particular owner and were fed by local people. Such individuals brought these animals to the Queen Saovabha Memorial Institute (QSMI), Bangkok, Thailand, for observation after they had bitten someone. These animals displayed no abnormal signs when first brought in. None had a history of vaccination. Once the animals started having abnormal signs, saliva samples were sent for rabies confirmation by nucleic acid sequence–based amplification (NASBA) as previously described . While waiting for NASBA results, which usually took 3–4 hours, these animals were closely observed by a veterinarian experienced in rabies infection [VT] for signs indicative of furious (aggression, biting/attacking the cage on seeing people approaching) or paralytic rabies (no or very minimal aggression with dropped jaw and hind limb paresis) . All animals that exhibited definite signs of either furious or paralytic rabies were humanely sacrificed as soon as the diagnosis was confirmed. None of them showed ambiguous clinical signs or changed from the furious to the paralytic form or vice versa. The duration between the first abnormal sign and autopsy in these animals was approximately 24 hours. Animals that were scheduled for MRI study were sedated prior to MRI examination, which was done within 24 hours after the first clinical sign. This procedure, including transfer to the MRI facility, MRI examination, and euthanasia took 4–6 hours, making the interval between the first clinical signs to autopsy in MRI-examined dogs approximately 36 hours. The period within 36 hours between the first sign and euthanasia for all animals in the study was based on the observation of 957 confirmed rabid dogs (two-third were furious) that were quarantined until death . The median survival time of rabid dogs was 4 days, with 25% of them died within 48 hours, and they usually lapsed into coma 12 hours before death.
During the 3 years of the specimen collection phase, there were 124 dogs observed at the Quarantine and Diagnostic Unit of the QSMI and 41 of them were determined to have rabies infection. 10 rabid dogs (5 furious and 5 paralytic) that met the criteria set above were considered to be in the early stage of the disease. All animals were conscious prior to euthanasia, except for the 4 (2 furious and 2 paralytic) that were sedated for imaging purposes. The results of the MRI studies have been previously published . Brains were available for pathological examination in all 10 dogs and spinal cords in 6 dogs (3 furious and 3 paralytic).
The diagnosis of rabies was confirmed by the presence of rabies antigen and viral nucleocapsid RNA in the brain by a direct fluorescent antibody test and by NASBA . All tissues were initially stored at −80°C immediately upon sacrifice. Samples for histology were taken immediately after thawing, fixed in 10% buffered formalin, routinely processed and embedded in paraffin wax. Brain and spinal cord tissue was collected from 11 sites in the brain and 4 in the spinal cord. A canine brain transaction atlas  was used to identify the region of interest as follows: frontal tip, parietal lobe (level 2), temporal lobe (level 5), occipital tip, cerebellum (vermis), caudate nucleus (level 2), hippocampus (including CA1–4 regions) (level 5), thalamus (level 4), midbrain (level 7), pons (level 12), medulla (level 17), and spinal cord (cervical, thoracic, lumbar, and sacral levels). For histologic examination, one section was taken from each brain and brainstem region and 3 from each spinal cord level. Samples from the 11 brain regions and from each spinal cord level were submitted for RABV RNA study as described below.
Extraction of RNA, reverse transcription, and RABV quantification by TaqMan real-time polymerase chain reaction (PCR) was done as previously described . The rabies viral load (N gene) was expressed as the copy number of rabies per μg total RNA. The levels of viral RNA in the different brain regions in 8 dogs (4 of each clinical form) have previously been published . In the present study, 2 more cases (one of each clinical form) and spinal cord tissue (6 cases, 3 of each clinical form) were included for analysis.
Indirect immunoperoxidase staining was carried out on four-micrometer-thick sections using an automated stainer (Ventana Benchmark LT, Tucson, USA). All tissue sections were cut by the same technologist using the same microtome. All steps of the immunostaining procedure were performed in a closed system using identical settings between runs, in order to maintain consistency in immunostaining. This system includes blocking with normal serum of the same species as the primary antibody, prior to application of the primary antibody. Primary antibodies included a polyclonal anti-rabies nucleocapsid antibody (Bio-Rad; Marnes-la-Coquette, France) at a dilution of 1:80, a polyclonal anti-CD3 antibody (DakoCytomation; Glostrup, Denmark at a dilution of 1:200), and a polyclonal anti-CD20 antibody (Abnova; CA, USA at a dilution of 1:100). Both CD3 and CD20 antibodies are directed against human epitopes but cross react some other mammalian species including dog. Although the anti-rabies nucleocapsid antibody was conjugated to FITC, fluorescence was not used for detection, and a secondary reagent was required for signal detection by light microscopy. The other two primary antibodies were unconjugated. The detection is done by the Ventana Ultraview technology that uses a multimeric secondary reagent. The negative control for each antibody consisted of omission of the primary antibody.
Semiquantitative analysis was done on the sections immunostained for rabies antigen. The amount of rabies antigen in neurons for each anatomical site was scored by a pathologist [SS] on a scale of 0 to 4+ as follows: 0 = negative, 1+ = 1-25% positive neurons, 2+ = 26-50%, 3+ = 51-75%, and 4+ > 75%. Analysis was performed without knowledge of the clinical data.
Quantitative analysis was performed using the immunostained slides for rabies antigen, which were captured in color by a digital camera (Nikon DXM1200F) attached to a microscope (Nikon ECLIPSE 80i) (Nikon Instech Co., Ltd., Japan) at 400× magnification. For each slide of the cerebral cortices, caudate nucleus and thalamus; the area with maximum RABV antigen was selected and neurons were digitally captured. Neurons in all layers of the cerebral cortices were included for analysis. Consecutive fields of the CA1 through CA4 regions were selected for the hippocampus, and the Purkinje cell layer for the cerebellum. All groups of neurons in the brainstem were captured, and only the gray matter was selected for all spinal cord levels. Each field covered an area of 0.0705 mm2 and from 10 to 20 fields (0.7-1.41 mm2) were captured from each slide. All neurons in each field were manually outlined using Adobe Photoshop. Quantitative assessment was performed by a computer-aided system (Cell Image Analyzer) (Figure 1) as previously described . This system measures the amount of chromogen signal regardless of the pattern of staining (e.g., granular vs. diffuse). Parameters assessed included: (1) percentage of positive neurons, (2) average antigen-positive area in positive neurons, and (3) average antigen-positive area per neuron [parameter 1 × parameter 2 ÷ 100]. The first parameter indicates how many neurons are affected while the others indicate the extent of involvement. The average number of neurons counted per dog was: cerebral cortices, 1881.6; cerebral midline (caudate and thalamus) and hippocampus, 782.6; cerebellum, 61.9; brainstem, 382.2; and spinal cord, 565.4.
Quantitative analysis of rabies viral antigen. Following immunostaining for rabies antigen (A), all neurons in each captured image are manually outlined (B) and analyzed by computer software. The three antigen parameters assessed are: (1) percentage of positive neurons, (2) average antigen area in positive neuron, and (3) average antigen area per neuron. (A and B, immunoperoxidase, original magnification X400).
Semi-quantitative analysis was performed on four-micrometer-thick sections of all paraffin blocks stained with hematoxylin and eosin. The inflammatory response in each anatomical site was scored by a pathologist [SS] on a scale of 0 to 3+ as follows: 0 = no inflammation, 1 = mild inflammation (mild microglial activation without formation of microglial nodules and/or mild perivascular lymphocytic cuffing), 2 = moderate inflammation (microglial activation with formation of microglial nodules and/or moderate lymphocytic perivascular cuffing), and 3 = marked inflammation (microglial activation with formation of microglial nodules with extension of microglia aggregates outside nodules, and/or marked perivascular lymphocytic cuffing with aggregates of lymphocytes outside the perivascular area). The degree of perivascular lymphocytic cuffing was defined by the number of layers of lymphocytes around vessels: mild, 1–2 cells; moderate, 3–4 cells, and marked, 5 cells or more.
Quantitative analysis was done on all CD3-immunostained slides. Images were digitally captured in and around the area of perivascular lymphocytic cuffing. 10 to 20 fields (0.7-1.41 mm2) were captured from each slide. Positive cells were manually counted, and the density of positive cells (the number of cell per 0.0705 mm2) calculated. CD20-immunostained slides were not quantitated since very few positive cells were found. The assessments were done without knowledge of the clinical data.
In quantitative assays, data from all fields were used to generate the average and SD. RABV antigen parameters and amounts of RNA were compared within the same group of animals at different CNS regions and between the different groups (furious versus paralytic), using the Mann–Whitney U test for a one-tailed test. Density of T cells was also compared between the different groups. A one-sided test was used because the semiquantitative analyses showed a trend towards a greater antigen burden in furious rabies (see Table 1) and towards greater inflammation in paralytic rabies (see Table 2). Statistical analyses were performed using Statistical Package for Social Science software (SPSS version 17.0, SPSS Inc., Chicago, IL, USA). A result was considered statistically significant at p < 0.05.
The semiquantitative results for number of neurons positive for RABV antigen are presented in Table 1, and the quantitative results for viral antigen parameters RNA amount according to major CNS regions of individual dogs are summarized in Table 3 and depicted in Figure 2. Both semiquantitative and quantitative studies show similar trends. RABV antigen-positive neurons at CNS regions were found most abundantly in the spinal cord followed by brainstem, cerebellum, cerebral midline structures (caudate, thalamus), hippocampus, and cerebrum. A significantly greater percentage of RABV-positive neurons was found in the spinal cord, as compared to the cerebrum and cerebral midline structures. The same trend was observed with the other viral antigen parameters (average antigen-positive area in positive neurons and average antigen-positive area per neuron). The largest quantities of viral mRNA were, however, observed in the cerebral midline structures and hippocampus, followed by brainstem. There was no significant difference in viral RNA levels between the spinal cord, brainstem, cerebellum, thalamus-caudate, hippocampus, and cerebrum. Prominent inflammation was noted in the brainstem (Table 2 and Figure 3).
Underlined P values indicate significant difference.
Rabies viral antigen and RNA pattern in paralytic and furious rabies. Rabies antigen parameters and RNA levels are shown for the major CNS regions. Each column color represents a single animal. Note that rabies viral antigen and RNA in one paralytic dog (dark blue column) was found exclusively in the spinal cord. RABV RNA is expressed in copies/μg total RNA × 108. RABV = rabies virus; Ag = Antigen; SP = spinal cord; BS = brainstem; CBL = cerebellum; CTH = caudate, thalamus, and hippocampus; and CB = cerebrum. No spinal cord tissue was available for paralytic rabies cases # 3 and 5 and furious rabies cases # 1 and 2 (refer to SP bars).
Brainstem inflammation in canine rabies. Striking inflammation (A) with a dense T cell infiltrate (C) in brainstem of paralytic rabies is shown, in comparison with a mild inflammatory response (B) and fewer T cells (D) in furious rabies. (A and B, hematoxylin and eosin stain, original magnification X 400; C and D, CD3 immunostain, original magnification X 400).
The semiquantitative results for the number of neurons positive for RABV antigen are presented in Table 1, and the quantitative results for viral antigen parameters and RNA amount according to major CNS regions of individual dogs are summarized in Table 4 and depicted in Figure 2. Similar to paralytic rabies, the spinal cord contained the most numerous RABV antigen-positive neurons, followed by brainstem, cerebellum, cerebral midline structures (caudate, thalamus), hippocampus, and cerebrum (Table 4). The percentage of RABV-positive neurons in the spinal cord was significantly greater than in the brainstem and brain regions. A similar pattern was noted with all three antigen parameters. As with paralytic rabies, the largest quantities of viral RNA were observed in the cerebral midline structures and hippocampus, followed by brainstem. Viral RNA level in the brainstem was significantly greater than in the cerebellum. Inflammation was generally mild throughout the entire neuraxis (Table 2).
The percentage of RABV antigen-positive neurons was significantly greater in the furious form compared to the paralytic form in the frontal (p = 0.038) and occipital (p = 0.029) lobes (Figure 4A). Viral antigen was ubiquitous in the brainstem neurons of both clinical forms with no significant difference. Viral antigen was widespread in the spinal cord of both forms, with a significantly greater percentage noted in the furious form at the cervical (p = 0.025), lumbar (p = 0.025), and sacral (p = 0.025) levels. This quantitative result correlated with the trend noted by semiquantitative assessment (Table 1).
Rabies viral antigen and RNA pattern comparison between paralytic and furious rabies. Rabies antigen parameters (A-C) and RNA levels (D) are shown for all CNS regions examined. Blue and red bars refer to paralytic and furious rabies, respectively. S = sacral cord, L = lumbar cord, T = thoracic cord, C = cervical cord, M = medulla oblongata, P = pons, Mb = midbrain, Cb = cerebellum, Th = thalamus, Cd = caudate nucleus, H = hippocampus, O = occipital lobe, Tm = temporal lobe, Pr = parietal lobe, and F = frontal lobe. The viral RNA is expressed as the copy number of rabies/µg total RNA × 108. Asterisks indicate significantly greater RABV antigen or RNA burden in the furious subtype compared to the paralytic subtype.
The area of RABV antigen in positive neurons was greater in the occipital lobe (p = 0.038), hippocampus (p = 0.023), caudate nucleus (p = 0.038), thalamus (p = 0.025), midbrain (p = 0.014), medulla (p = 0.043), and cervical cord (p = 0.025) of the furious form compared to the paralytic form (Figure 4B).
The area of RABV antigen per positive neuron was significantly greater in the occipital lobe (p = 0.037), midbrain (p = 0.023), medulla (p = 0.014), and cervical spinal cord (p = 0.024) and lumbar spinal cord (p = 0.025) of the furious form compared to the paralytic form (Figure 4C).
Significantly greater levels of viral RNA were demonstrated in all cerebral neocortices (p = 0.004, 0.014, 0.038, 0.008, for the frontal, parietal, temporal, and occipital lobes, respectively), thalamus (p = 0.024), midbrain (p = 0.024), and medulla (p = 0.024) in the furious form compared to the paralytic form (Figure 4D). The amount of rabies viral RNA in the spinal cord was comparable in both forms.
More intense inflammation (2 + to 3+) was noted in the brainstem of the paralytic form compared to the furious form (Table 2). The inflammatory reaction in the brain and spinal cord was not remarkable in either form (0 to 1+). CD3-positive T lymphocytes strikingly outnumbered CD20-positive B-cells. The number of T-cells paralleled the inflammatory scores. The density of T-cells (the number of cell/0.0705 mm2) at the brainstem was significantly greater in paralytic rabies (Figure 3) compared to the furious subtype (70.7 ± 7.41 vs 13.04 ± 2.43, p = 0.004). No significant difference in T cell density was noted between the two clinical forms in other parts of the CNS.
It is intriguing that rabies in humans and dogs can manifest in either furious or paralytic forms. The observation that the same rabid dog transmitted paralytic rabies to one human and furious to another human  suggests that host response may play a role in determining the clinical subtype. Previous neuropathological studies in rabies-infected humans have not been able to demonstrate differences between these two clinical subtypes . Immunohistochemistry has been employed in limited studies to detect rabies nucleocapsid antigen, and assessment done in a semi-quantitative fashion. In terms of RABV antigen abundance, the thalamus, basal ganglia, and brainstem have been shown to be preferentially involved over the cerebral hemispheres regardless of the clinical forms . A study in rabid dogs also showed greater RABV antigen burden in the brainstem compared to supratentorial structures . However, in this animal study, only the furious form was included and no spinal cord was available for analysis.
In the present study, semi-quantitative and quantitative assessment of RABV antigen was performed in both furious and paralytic forms at the early stage of the disease, and the spinal cord was studied for the first time in canine rabies. The viral antigen burden in both clinical forms was greatest in the spinal cord, followed by brainstem, and then the brain. This caudal to rostral trend of decreasing RABV antigen abundance was shown by semi-quantitative assessment, and quantitative assessment of all three parameters (% positive neurons, % antigen area in positive neuron, and % antigen area per neuron). Greater viral antigen amount was found in the furious form in most CNS regions including the spinal cord. At first, this latter finding might be viewed as paradoxical since one might expect the viral load in the spinal cord would be greater in the paralytic form. However, it is known that the clinical weakness in the paralytic form is due to peripheral nerve dysfunction, rather than the presence of RABV in the spinal cord . Of diagnostic interest, in one dog with the paralytic form, RABV antigen was found only in the spinal cord. Therefore, for making a diagnosis of rabies in a dog using an antigen detection method, a spinal cord specimen should be included, especially when the suspected animal is sacrificed shortly after the onset of illness or when not yet unconscious from the disease.
RABV RNA was distributed almost uniformly in all CNS regions by the early stage of the disease, regardless of the clinical subtype, reflecting the rapidly aggressive nature of rabies infection. The larger viral RNA load in all CNS sites in the furious form compared to paralytic form may explain the shorter survival period in the furious form and perhaps the more dominant cerebral symptoms seen in both human and canine rabies. Although it has been shown that RABV RNA is synthesized within a cage of nucleocapsid protein in the Negri body-like structures , the distribution of viral RNA in our study did not mirror that of rabies nucleocapsid protein. Whereas there were significant differences in the amount of RABV antigen at different levels of the CNS, this was generally not the case for viral RNA. While the brainstem, thalamus, caudate and hippocampus contained higher copy numbers of RABV RNA compared to other regions, this was not statistically significant. The cerebellum was the only site to have significantly less viral RNA compared to the brainstem and only in the furious form. Furthermore, the spinal cord, where RABV antigen was most abundantly found, contained relatively low amounts of viral RNA.
The discrepancy between RABV antigen and RNA amounts reflects the dynamic nature of viral transcription and replication and could be related to differences in production and degradation of RABV mRNA as distinct from viral antigen, and/or host factors controlling of transcription such as miRNAs. Different CNS regions may vary in resistance to viral propagation and cytolytic effects. In support of this hypothesis, rat spinal cord motor neurons are more resistant in vitro to RABV-induced cytolysis in a fixed virus model . Other host factors may also play important roles in determining the clinical subtype of rabies infection. A greater immune response occurs with paralytic rabies, as evidenced by cytokine mRNA transcripts in the brain of dogs .
A noteworthy finding in our study is that the greatest degree of inflammation was found in the brainstem of dogs with the paralytic form. Similar to other viral infections, T-cells predominated over B-cells in the inflammatory reaction. These findings correlated with the imaging findings showing more abnormal signals by MRI, indicating a greater degree of inflammation, in dogs with the paralytic form compared to dogs with the furious form, particularly at the brainstem level . Moreover, interleukin-1β and interferon-γ mRNAs were found exclusively in the paralytic form . Our findings also correlate with a mouse model study in which minocycline-treated mice showed reduced inflammation and increased disease severity when infected with SAD-D29, an attenuated strain of rabies . The heightened inflammatory response in the canine brainstem in paralytic rabies is in line with the impairment of neural tract integrity recently demonstrated by increased FLAIR (fluid attenuated inversion recovery) signal on MRI and decreased fractional anisotropy values on diffusion tensor imaging . Disruption of axoplasmic flow by inflammation in the brainstem could potentially retard viral propagation, particularly towards the cerebral hemispheres in the situation of paralytic rabies. This, in turn, could result in a diminished viral load in the brainstem and cerebrum. Why the maximum degree of inflammation was seen in the brainstem is not known. This may reflect the site of initial entry into the CNS by the RABV, since most dogs would be bitten in the neck region, and the virus would then migrate along the cranial nerves to enter the CNS at the level of the brainstem. However, we cannot say with certainty at which sites the dogs in our study were bitten, and thus this remains speculation. We also recognize the small number of animals available for study limits the strength of any conclusions we might reach. Carrying out larger studies in dogs in a controlled laboratory setting would overcome some of these limitations, and would also allow a comparison of clinical and pathologic findings between immune-competent and immune-deficient dogs. However, experimental studies for rabies are generally done in rodents rather than dogs, in part for ethical reasons. There also arise concerns over which virus to use in the laboratory setting. Wild-type virus is difficult to control with respect to the amount of virus to inoculate in order to create a reproducible incubation period, clinical picture, and mortality rate. Laboratory strains of RABV do not duplicate the natural infection. For example, elimination of RABV virulent strains is impeded by apoptosis of migratory inflammatory cells, whereas laboratory RABV strains produce neuronal apoptosis that is not seen with the wild-type virus [22, 26–28]. As well, other studies have shown laboratory strains are associated with activation of the host immune response, whereas there is very little response to wild-type virus . Therefore, there are distinct advantages to studying rabies in the natural setting.
Host factors may not be the only explanation for the two clinical subtypes of rabies. We were previously unable to detect any specific differences in the glycoprotein, phosphoprotein and nucleocapsid genes of RABV isolated from the furious and paralytic forms in humans and animals . The glycoprotein gene was found to have minor sequence variations, and cloned sequences of the viral population derived from a single rabies-infected dog showed minor substitutions at both nucleotide and amino acid levels . However, both reports analyzed viruses isolated only from one part of the brain. Hence, it remains possible that viral genetic polymorphisms might play a role in determining the clinical manifestations and influencing the host inflammatory and immune responses.
Our results provide preliminary evidence that rabies virus appears to have a greater ability for dissemination in the furious subtype compared to the paralytic. The lower amount of rabies viral antigen and RNA in several CNS regions of dogs with the paralytic form is associated with conspicuous inflammation at the brainstem. Different host inflammatory responses might account for the two clinical forms of rabies. This inflammatory reaction could lead to impaired viral dissemination towards the CNS, particularly into the cerebral hemispheres in paralytic rabies, and might explain the longer survival time compared to the furious form. Additional research including controlled laboratory studies is needed to examine this hypothesis.
Research was in part sponsored by the Thailand Research Fund (DBG5180026); Integrated Innovation Academic Center (IIAC), Chulalongkorn University Centenary Academic Development Project; the Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission (HR1160A-55); and the Naval Health Research Center under Cooperative Agreement Number W911NF-11-2-0041. The views and conclusions contained in this document are those of the authors and should not be interpreted as representing the official policies, either expressed or implied, of the Army Research Laboratory or the U.S. Government. The U.S. Government is authorized to reproduce and distribute reprints for Government purposes notwithstanding any copyright notation herein.
SS designed the study, performed histopathological examination, and participated in data analysis. NT carried out immunohistochemical stainings and image analysis. PP designed computer program for image analysis. AJ performed real-time PCR, and NI performed statistical analysis. VT took care of the animal and collected samples. All authors, including SW, PT, and TH, involved in drafting the manuscript and approved the final version.
Global Alliance for Rabies Control. http://www.rabiescontrol.net.


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Due to the ease of access to journals, electronic books and expert opinions available online, many writers find themselves citing Web sources in research papers. The sixth edition of the "Publication Manual of the American Psychological Association" establishes specific guidelines you should follow when using Internet sources in your APA writing.
In-text citations for online publications follow the same rules as those for print references: author's last name and publication year. The references entry should help your reader find that source, so a basic Web page entry begins with the author, last name followed by first initial with a comma in between. After a period, the date appears, in parentheses, with a period at the end. Then give the title of the page, capitalizing only the first word and any proper nouns, a period and "Retrieved from" -- without the quotation marks -- followed by the entire URL with no punctuation at the end. Such an entry looks like this:
Jones, K. (2013). Happiness is a bowl of peanuts. Retrieved from http://www.nuts.com/home
Electronic books follow this same format except the title of the book appears in italics if no DOI -- or Digital Object Identifier -- exists. Do not add a retrieval date unless you cite material that typically changes often, such as in a Wiki.
References page entries for online periodicals include more detail about the publication in which they appear. They start with the author, last name followed by first initial with a comma between and a period at the end. The date appears in parentheses next, followed by a period. The title of the article comes next, capitalizing the first word and proper nouns only, with a period at the end. Italicize the name of the periodical and the volume with a comma between. After a comma, give the print page numbers and a period. End the entry with the online reference such as the URL. An entry looks like this example:
Jones, K. (2013). Happiness is a bowl of peanuts. Journal of Sociology, 31, (italicized) 32-25. Retrieved from http://www.nuts.com/home
If the journal also has an issue number, put it in parentheses immediately after the volume, before the comma. For instance, 31(2) indicates volume 31, issue number 2. The issue is not italicized.
The DOI system assigns alphanumeric tags to online publications, particularly periodicals, to make them easier to find. APA requires the inclusion of the DOI in the references entry when you find one, typically at the top or bottom of the first page of an article. The DOI replaces the URL in a references page entry, following "doi:" -- without the quotation marks -- and the string. An entry looks like this example:
Jones, K. (2013). Happiness is a bowl of peanuts. Journal of Sociology, 31, (italicized) 32-25. doi: 10/8486.88609
Other kinds of sources, such as discussion or forum posts, follow slightly different structure on the references page. The entry begins with the poster's name, in inverted form just like an author if given. If not, use the screen name. After a period, give the specific date of the post in parentheses, using year-month-day order, putting a comma after the year. Place a period after the parentheses. The poster responds to an article, so type "Re:" -- without the quotation marks -- to indicate this is the subject the person was writing about followed by the name of the article or discussion. An explanation of the type of post follows in square brackets, such as [Web log comment]. The entry ends with the URL. A post reference looks like this:
Jefferson, L. (2013, April 25). Re: The nuts and the bolts of happiness and its relation to peanuts [Online forum comment]. Retrieved from http://www.nuts.com/forum/april
Sweet, Kristie. "APA Format for Online Referencing." , https://penandthepad.com/apa-format-online-referencing-2021.html. Accessed 17 July 2019.
Sweet, Kristie. (n.d.). APA Format for Online Referencing. . Retrieved from https://penandthepad.com/apa-format-online-referencing-2021.html
Sweet, Kristie. "APA Format for Online Referencing" accessed July 17, 2019. https://penandthepad.com/apa-format-online-referencing-2021.html
Publication Manual of the American Psychological Association (6th edition); American Psychological Association
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Watch: Mysterious "Levitating" Cars at Busy Intersection in China
HenSpark / Latest Stories / Bizarre / Watch: Mysterious "Levitating" Cars at Busy Intersection in China
A video taken at a busy intersection in the city of Xingtai, central China has gone viral because of bizarre circumstances that made three vehicles appear to levitate all of a sudden even while the rest of the vehicles and the people in the video did not look affected at all.
In this video, nothing appears to be amiss at first. There were several vehicles on the streets leading to the intersection, with pedestrians waiting to cross the streets once the traffic lights change.
Then, all of a sudden, a van's rear end appears to levitate while its front section remained on the ground.
The vehicle adjacent to the van soon followed suit, with its front section moving up but the back remained rooted on the ground.
While the first vehicle "levitated" even higher and eventually fell on its side, the second vehicle also moved in the same direction and the two hit each other.
Meanwhile, at the right side of the video frame, the back of another car also appeared to levitate for a moment but this was not as incredible as what happened to the two vehicles that crashed on each other at the left side of the frame.
The mysterious video went viral because no one could seem to provide the best explanation why this bizarre event happened.
Netizens put forth various explanations, including unbelievable theories like the levitation was the work of aliens who made a temporary force field over the cards.
Some netizens also said this supposed CCTV video was simply faked because it was made in China and could have been tweaked to make the vehicles appear real.
Well, the vehicles did appear real and the video, too. So, what could have possibly made those cars move in bizarre fashion?
Thankfully, the mystery was solved by the Chinese authorities who handled the incident.
According to reports from Chinese media, there really was not much mystery in what happened with the cars.
It turns out that the large vehicle on the right side, next to the affected car, is a street sweeper.
A fallen steel cable was caught up in the street sweeper's brush.
As the street sweeper pulled the line, the vehicles who were on top of the line got pulled up, making them appear to levitate without any explanation.
Because of the grainy quality of the CCTV video, the steel line was not clear to see.
So, there you have it. Mystery solved.
(via Daily Mail UK)
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{{Ficha de competición deportiva
|nombre          = Abierto de Australia 2020
|nombre_original = Australian Open 2020
|deporte         = Tenis
|tipo            = Grand Slam
|recinto         = Melbourne Park
|ubicación       = Melbourne
|país            =
|fecha           = 20 de enero - 2 de febrero
|superficie      = Dura (GreenSet)
|edición         = 108.ª
|cuadro          = 128S/128Q/64D/16Q/48X
|premio          = A$D 71 000 000
|organizador     = Federación Internacional de Tenis
|individual masculino =  Novak Djokovic
|individual femenino  =  Sofia Kenin
|dobles masculino     =  Rajeev Ram Joe Salisbury
|dobles femenino      =  Tímea Babos Kristina Mladenovic
|dobles mixto         =  Barbora Krejčíková Nikola Mektić
|anterior        = 2019
|actual          = 2020
|siguiente       = 2021
|página web      = http://ausopen.com/
}}
El Abierto de Australia 2020 fue un torneo de tenis celebrado en las pistas de superficie dura del Melbourne Park, situado en Melbourne (Australia). El campeonato se celebró entre el 20 de enero y el 2 de febrero de 2020. Se trató de la 108.ª edición del Abierto y el primer torneo de Grand Slam de 2020. Contó con eventos individuales y por parejas de tanto masculinos, como femeninos, además de dobles mixtos, júnior y silla de ruedas.

Novak Djokovic y Naomi Osaka fueron los campeones defensores en las ramas de individuales masculinas e individuales femeninas, respectivamente.

Fue la primera edición del Abierto de Australia que se jugó en GreenSet, un tipo de cancha dura fabricado por la compañía GreenSet Worldwide. GreenSet es el tercer tipo de superficie dura usada por el Australian Open.

Torneo

El Abierto de Australia 2020 fue la edición 108va del torneo y se celebró en Melbourne Park en Melbourne, Victoria, Australia.

El torneo fue dirigido por la Federación Internacional de Tenis (ITF) y es parte del calendario 2020 del ATP Tour y de la WTA Tour bajo la categoría de Grand Slam. El torneo consiste en dos cuadros de las modalidades de individuales y dobles tanto para hombres como para mujeres, así mismo como un evento de dobles mixtos. Hay cuadros de individuales y dobles también para jugadores juniors, jóvenes y señoritas (jugadores menores de 18). Hay también competiciones en individuales y dobles para varones y damas en Tenis adaptado.

El torneo fue jugado en pistas duras de cemento alrededor de 25 pistas de tenis en el complejo, los tres principales estadios son la  Rod Laver Arena, la Melbourne Arena y la Margaret Court Arena. La 1573 Arena (anteriormente Pista 2) fue promovida a una pista principal.

El torneo contó con una característica especial, debido a que estaba siendo amenazado su realización, debido a los incendios forestales que estaban azotando a Australia en septiembre del año pasado, por lo que con el pasar del tiempo se logró realizarlo sin mayores contratiempos.

 Puntos y premios
 Distribución de puntos
 Séniors

 Tenis adaptado

* Categoría específica de tenis en silla de ruedas

 Júniors

</center>

 Premios monetarios
El Abierto de Australia en su edición 2020 ha incrementado los premios en metálico a un máximo histórico de 71 millones de dólares australianos, un 13.6% más que en 2019.

 Notas

 Actuación de los jugadores en el torneo
 Individual masculino

 Individual femenino

Sumario
 Día 1 (20 de enero)
 Cabezas de serie eliminados:
 Individual masculino:  Denis Shapovalov [13],  Borna Ćorić [25]
 Individual femenino:  Sloane Stephens [24],  Barbora Strýcová [32]
 Orden de juego

 Día 2 (21 de enero)
 Cabezas de serie eliminados:
 Individual masculino:  Félix Auger-Aliassime [20],  Jo-Wilfried Tsonga [28]
 Individual femenino:  Johanna Konta [12],  Markéta Vondroušová [15],  Amanda Anisimova [21],  Anastasija Sevastova [31]
 Orden de juego

 Día 3 (22 de enero)
 Cabezas de serie eliminados:
 Individual masculino:  Matteo Berrettini [8],  Grigor Dimitrov [18],  Benoît Paire [21],  Daniel Evans [30],  Hubert Hurkacz [31]
 Individual femenino:  Aryna Sabalenka [11],  Petra Martić [13],  Dayana Yastremska [23]
 Dobles masculino:  Kevin Krawietz /  Andreas Mies [3]
 Dobles femenino:  Nicole Melichar /  Yifan Xu [5],  Lyudmyla Kichenok /  Zhaoxuan Yang [14]
 Orden de juego

 Día 5 (24 de enero)
 Cabezas de serie eliminados:
 Individual masculino:  Stefanos Tsitsipas [6],  Roberto Bautista [9],  Guido Pella [22],  Dušan Lajović [24]
 Individual femenino:  Naomi Osaka [3],  Serena Williams [8],  Madison Keys [10],  Ekaterina Alexandrova [25],  Elena Rybakina [29]
 Dobles masculino:  Pierre-Hugues Herbert /  Nicolas Mahut [1],  Jean-Julien Rojer /  Horia Tecău [8],  Raven Klaasen /  Oliver Marach [9],  Jürgen Melzer /  Édouard Roger-Vasselin [12]
 Dobles femenino:  Yingying Duan /  Saisai Zheng [9],  Ellen Perez /  Samantha Stosur [12]
 Orden de juego

 Día 6 (25 de enero)
 Cabezas de serie eliminados:
 Individual masculino:  David Goffin [11],  Karen Jachanov [16],  John Isner [19],   Pablo Carreño [27],  Taylor Fritz [29]
 Individual femenino:  Karolína Plíšková [2],  Elina Svitolina [5],  Belinda Bencic [6],  Donna Vekić [19]
 Dobles masculino:  Łukasz Kubot /  Marcelo Melo [2],  Wesley Koolhof /  Nikola Mektić [5],  Jamie Murray /  Neal Skupski [14],  Máximo González /  Fabrice Martin [15]
 Dobles femenino:  Květa Peschke /  Demi Schuurs [8]
 Orden de juego

 Día 7 (26 de enero)
 Cabezas de serie eliminados:
 Individual masculino:  Fabio Fognini [12],  Diego Schwartzman [14]
 Individual femenino:  Alison Riske [18],  Maria Sakkari [22],  Qiang Wang [27]
 Dobles masculino:  John Peers /  Michael Venus [7],  Mate Pavić /  Bruno Soares [10]
 Dobles mixto:  Barbora Strýcová /  Marcelo Melo [1]
 Orden de juego

 Día 10 (29 de enero)
 Cabezas de serie eliminados:
 Individual masculino:  Rafael Nadal [1],  Stan Wawrinka [15]
 Individual femenino:  Anett Kontaveit [28],  Anastasia Pavlyuchenkova [30]
 Dobles femenino:  Barbora Krejčíková /  Kateřina Siniaková [4],  Hao-Ching Chan /  Latisha Chan [7]
 Orden de juego

 Bajas femeninas notables

 Cabezas de serie dobles

 Dobles masculino

 Dobles femenino

<center>

 Dobles mixto

</center>

 Notas

 Invitados

 Individual masculino
  Alex Bolt
  Hugo Gaston
  Michael Mmoh
  Andrew Harris
  Tatsuma Itō
  Christopher O'Connell
  Marc Polmans
  John-Patrick Smith

 Individual femenino
  Lizette Cabrera
  Na-Lae Han
  Priscilla Hon
  Pauline Parmentier
  Arina Rodiónova
  María Sharápova
  Astra Sharma
  Coco Vandeweghe

 Dobles masculino
  Alex Bolt /  Matthew Ebden
  James Duckworth /  Marc Polmans
  Blake Ellis /  Alexei Popyrin
  Andrew Harris /  Christopher O'Connell
  Lleyton Hewitt /  Jordan Thompson
  Ji-Sung Nam /  Min-Kyu Song
  Max Purcell /  Luke Saville

 Dobles femenino
  Destanee Aiava /  Lizette Cabrera
  Alexandra Bozovic /  Amber Marshall
  Jaimee Fourlis /  Arina Rodiónova
  Priscilla Hon /  Storm Sanders
  Maddison Inglis /  Kaylah McPhee
  Ya-Hsuan Lee /  Fang-Hsien Wu
  Jessica Moore /  Astra Sharma

 Dobles mixto
  Monique Adamczak /  David Vega Hernández
  Jessica Moore /  Matthew Ebden
  Jeļena Ostapenko /  Leander Paes
  Ellen Perez /  Luke Saville
  Arina Rodiónova /  Andrew Harris
  Storm Sanders /  Marc Polmans
  Astra Sharma /  John-Patrick Smith
  Belinda Woolcock /  Blake Mott

Clasificación

 Clasificación masculina

  Dennis Novak
  Tallon Griekspoor
  Christopher Eubanks
  Elliot Benchetrit
  Mario Vilella Martínez
  Mohamed Safwat
  Ilya Ivashka
  Quentin Halys
  Marco Trungelliti
  Norbert Gomboš
  Daniel Elahi Galán
  Pedro Martínez
  Max Purcell
  Alejandro Tabilo
  Ernests Gulbis
  Peter Gojowczyk

 Perdedores afortunados
  Evgeny Donskoy
  Prajnesh Gunneswaran
  Jozef Kovalík
  Lorenzo Giustino

 Clasificación femenina

  Ann Li
  Nao Hibino
  Johanna Larsson
  Barbora Krejčíková
  Anna Kalínskaya
  Kaja Juvan
  Leylah Annie Fernandez
  Shelby Rogers
  Martina Trevisan
  Caty McNally
  Monica Niculescu
  Liudmila Samsonova
  Greet Minnen
  Elisabetta Cocciaretto
  Harriet Dart
  Antonia Lottner

Ranking protegido
Los siguientes jugadores entraron directamente al cuadro principal al usar su ranking protegido:

 Individual masculino
  Yen-Hsun Lu (PR 71)
  Mackenzie McDonald (PR 83)
  Vasek Pospisil (PR 73)
  Cedrik-Marcel Stebe (PR 95)

 Individual femenino
  Catherine Bellis (PR 44)
  Kateryna Bondarenko (PR 85)
  Katie Boulter (PR 85)
  Anna Karolína Schmiedlová (PR 93)

 Campeones defensores

 Campeones
 Sénior
 Individual masculino  Novak Djokovic venció a  Dominic Thiem por 6-4, 4-6, 2-6, 6-3, 6-4

 Individual femenino  Sofia Kenin venció a  Garbiñe Muguruza por 4-6, 6-2, 6-2

 Dobles masculino  Rajeev Ram /  Joe Salisbury vencieron a  Max Purcell /  Luke Saville por 6-4, 6-2

 Dobles femenino  Tímea Babos /  Kristina Mladenovic vencieron a  Su-Wei Hsieh /  Barbora Strýcová por 6-2, 6-1

 Dobles mixto  Barbora Krejčíková /  Nikola Mektić vencieron a  Bethanie Mattek-Sands /  Jamie Murray por 5-7, 6-4, [10-1]

 Júnior
 Individual masculino  Harold Mayot venció a  Arthur Cazaux por 6-4, 6-1

 Individual femenino  Victoria Jiménez Kasintseva venció a  Weronika Baszak por 5-7, 6-2, 6-2

 Dobles masculino  Nicholas Ionel /  Leandro Riedi vencieron a  Mikołaj Lorens /  Kārlis Ozoliņš por 6-7(8-10), 7-5, [10-4]

 Dobles femenino  Alexandra Eala /  Priska Nugroho vencieron a  Živa Falkner /  Matilda Mutavdzic por 6-1, 6-2

 Tenis adaptado
 Individual masculino  Shingo Kunieda venció a  Gordon Reid por 6-4, 6-4

 Individual femenino  Yui Kamiji venció a  Aniek van Koot por 6-2, 6-2

 Quad individual  Dylan Alcott venció a  Andrew Lapthorne por 6-0, 6-4

 Dobles masculino  Alfie Hewett /  Gordon Reid vencieron a  Stéphane Houdet /  Nicolas Peifer por 4-6, 6-4, [10-7]

 Dobles femenino  Yui Kamiji /  Jordanne Whiley vencieron a  Diede de Groot /  Aniek van Koot por 6-2, 6-4

 Quad dobles  Dylan Alcott /  Heath Davidson''' vencieron a  Andrew Lapthorne /  David Wagner por 6-4, 6-3

Véase también

 Tenis en 2020
 Torneos ATP en 2020
 Torneos WTA en 2020

Referencias

redpajama_set_name: RedPajamaWikipedia


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Yesterday was officially the first day of Spring and if you're like us then you're more than ready for the sunshine and outdoors! While most of us dread doing yard work, there are a number of steps you can take now to prevent lawncare troubles. To save you some time, we have compiled a list of helpful tips for your preparation of spring weather!
Back to school can be one hectic time for busy parents. Getting back into the school routine can be plain hard. Whether it's sprinting to catch the bus or sitting in the school's drop-off lane with overtired kids, or picking that perfect outfit when laundry is overflowing the baskets, mornings can be especially difficult. We've compiled tips on how to transition back to our back-to-school schedules!
We are less than a week away from the Great American Total Solar Eclipse! According to NASA, more than 300 million people in the U.S. will be able to directly view this celestial phenomenon on Monday, Aug. 21! The path of totality will span 14 states. We have tips for you to have a great - and safe - eclipse viewing experience!
Dull, boring, and cluttered laundry rooms are a thing of the past! Have you ever walked into a friend's home and been completely jealous of their forward-thinking layout and beautifully designed laundry room? Well, we're here to help! These 5 easy tips are sure to make your laundry room go from drag to fab.
Are you seeing your friends and family post their incredible vacation photos on social media and feeling the urge to get away, too? But when you're tight on vacation days, or vacation funds, that can be a challenge. Or maybe you've already had your vacation, but still feel the need to get away? No worries though -- that's the perfect opportunity for a day trip! Here's our Top 5 Day Trips for each of the Carolinas and Virginia!
If you are in the market for a new home, you have a huge opportunity to be considerate of your planetary home sweet home by shopping for a house that is built with energy efficiency in mind. To celebrate Earth Day 2017, we explain how to use a HERS rating to determine exactly how "green" that new home is!
With the turn of spring comes springing cleaning and spring home maintenance. Here's a helpful list of what you should be watching for in your home!
As we all face the reality that we are now officially in winter, we have tips to help you safeguard your home from the winter season. Most of the tasks are simple and can be done yourself, but a few do require professional assistance. Investing time and energy now can help reduce your energy bills and save from maintenance related headaches in the future!
The path for Hurricane Matthew is still unpredictable but now is the time to make plans for this Category 4 hurricane. Please review these tips and suggestions to help keep you and your property safe and sound.
From smart phones to smart homes, our world is changing fast. Eastwood Homes provides its homeowners options for securing their new homes, and for making them smarter, too.
What does this hot spring market mean for buyers?
Low interest rates, an even lower supply of homes, an improving jobs outlook and rising house prices are swirling together to make for a tornado of activity between buyers and sellers as the spring market heats up.
Building a new home together can be stressful for any couple. In honor of this Valentines holiday, we speak to one newlywed couple who came through the endeaver with a stronger marriage and a beautiful home in which to start their lives.
To book an appointment, or not?
Searching for a new home can be complicated and time consuming. However, it doesn't have to be! Simply chatting with our internet team first can make your visit to the community even more productive and enjoyable. Here's how!
If your New Year's Resolutions include buying a new home, we have some tips to help you get moving! The homebuying process can be overwhelming, but taking the time to prepare for this milestone will make the process easier and more enjoyable.
Selling a first home can easily be just as exciting and emotional as buying your first home – and maybe even doubly so! Not only are you looking for your next home but you are also parting with your very first home, which oftentimes is filled with happy memories. We have some tips that can help be the difference in selling your home in three weeks or three months.
Some tips to prepare for rains, flooding and high winds and keep your family and home safe during this weekend's predicted storms.
If you're considering purchasing a new construction home, you may have questions about the steps in the building process. You may even be a little overwhelmed at the thought. However, that shouldn't be the case! We are here with these tips to provide the information you need that will make you feel more knowledgeable about the process!
After choosing a builder, homebuyers need to start thinking about a perfect floor plan for their needs. There are many options from which to choose, including single-story, two-story, and three-story homes. But sometimes, homebuyers get caught up in the numbers of square footages and bedrooms when they should be focusing on livability!
What Type of New Home Shopper Are You?
What type of home shopper are you? Eastwood Homes compares characteristics of new home shoppers to our canine companions!
Did you know that cleaning and organizing our homes can make us feel better, both while we are doing it and later on while we are enjoying the more organized space? However, spring cleaning can be a daunting task to plan for. So, instead of trying to do it all, why not make a strategic plan to do one area really well.
Now is the time to begin our spring yard maintenance if we want a lawn to be proud of when we begin enjoying the warmer weather in just a few weeks. By putting in the work to prepare your yard now, you will better be able to enjoy your yard for the next several months. Our blog shares a few maintenance tips for lawn care to help you get started!
The FHA has lowered its annual mortgage insurance premiums by 50 basis points, from 1.35 percent to 0.85 percent. Lower mortgage insurance premiums are going to allow people to afford more home on the same budget!
Winter Maintenance Tips and Tricks from Eastwood Homes to prepare your home for the upcoming season!
When it comes to purchasing a new home, timing is one of the most important factors. If you're a newly married couple looking for your first home or a single retiree looking for a place to down-size, timing is inevitably going to play a role in your purchase decision. Is it the right time to make a change? Once you've decided that you are ready to make a change in your living arrangements, you've completed half the battle! The other, and often times, more difficult half of the battle still awaits. Is now the right time to buy?
Dream Home vs. School District. Does it have to be a competition?
August is upon us! Although the heat in the Southeast is here to stay (at least a few more months), summer is officially winding to a close. In just a few short weeks, school-age children will be loading up their back packs, grabbing their books, and heading out the door to the first day of school! From now until June, they will spend the majority of their day learning, growing, and developing in varying school environments. Because of the time and importance devoted to education, school options can play an important role in the home buying decision!
Beach Bound! Where's Your Favorite Fun in the Sun Getaway Located?
Summer is here! The weather is hot and the days are long! If you live in the Southeast like we do, this means it's the time of year to hit the beach! If you live in a state where Eastwood Homes is building, then you know what I'm talking about!
Rates are still LOW, and many builders like Eastwood Homes have beautiful move-in-ready inventory homes available. It really is a great time to buy a new home, but you don't need to take our word for it. Check out the following article from U.S. News and World Report. The full article can be found here.


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text:
The Canterbury Journal
Home Review Where can you find the best pub Sunday lunch in Canterbury?
Where can you find the best pub Sunday lunch in Canterbury?
Alex Lister
The glorious Sunday lunch - a mainstay of British identity
It has to be one of the best things about living in the UK. The glorious Sunday roast enjoyed in a traditional English pub, with a roaring fire in winter and a sun-drenched garden in summer – that's correct, sun-drenched rather than just drenched.
So if you're hankering for a great Sunday lunch in a great pub, go and stuff yourself full of roast potatoes, Yorkshire puddings and all the rest at one of these top locations.
Foodies! Is the latest London culinary rage about to hit Canterbury?
From outfitter to pub and kitchen, the remarkable journey of a famous city centre spot
Ye Olde Beverlie
In a secret corner of Canterbury, away from the tourist throng, lies the beautiful "Old Bev". Built in the 16th century, the pub boasts a huge log fireplace, and a large garden which plays home to the resident bat and trap team.
Sunday roasts are generous in size, and reasonably priced. For anyone who fancies something different they also serve sizzling fajitas, but let's be honest – that's probably not why you're there.
The Coach & Horses
Another hidden gem can be found in Harbledown, a stone's throw from the London road roundabout and recently featured by the Journal's resident food critic. Set against a steep rising slope, the garden is terraced and offers great views over the valley.
Beside the garden lives the resident black pig. Great entertainment for the children – as long as they don't think too carefully about what they're eating. But sausages come from the supermarket don't they?
The food is excellent, and there are some great walks on your doorstep if you feel the need to work it off.
The Parrot
If you prefer something a little more central, the Journal recommends the Parrot, found on Church Lane in St Radigunds, just round the corner from the Mint Yard Gate. Known as Simple Simon's until 2008, it is believed to be the oldest pub in Canterbury, and is one of the oldest buildings in the city.
The nearby Dolphin and the Millers Arms are both notable pubs, and literally metres from the doorstep, but the Parrot just pips them to the post in our opinion.
The Sunday lunch is excellent, and the interior is quirky and charming. There's also a hidden courtyard garden for the summer.
The Dog Inn has rooms available for overnight stays
The Dog Inn
The recently renovated Grade II* listed public house in Wingham, is a particular treat. Located five miles outside Canterbury, it's situated directly opposite a handy bus stop which will return you to Canterbury in around 20 minutes (but check the frequency on a Sunday).
The historic building is everything a village pub should be and the Sunday roast won't disappoint.
And after a couple of glasses of wine, it's hard to stifle a titter over the slightly unfortunate name. Let's just say the Dog Inn in Wingham comes highly recommended.
The George and Dragon
Topping the list of the Journal's favourite pub roasts is the George in Fordwich. About an hour's walk alongside the river from Canterbury, visitors are welcomed by a long bar, a log fire, a garden overlooking the river, and probably the best Sunday roast to be found locally. It's not the cheapest, but boy is it worth it!
Considering the competition, this is quite a controversial boast, but whenever we've sampled the Sunday lunches, they've been nothing short of superb. This is truly the one to beat.
Previous articleYes, we're pothole central – but why is that?
Next articleFacing the barrel of a gun: How high-flying banker became Buddhist nun
https://www.alexlister.com
Alex Lister lives in Canterbury with his wife and children. He is a writer, political campaigner, digital communications professional and hospital governor.
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About The Canterbury Journal
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VIDEO: Fascinating footage shows city nearly 100 years ago
Roll up, roll up, the circus is in town!


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text:
Q: Unable to invoke Apex Class in lightning component for non-admin user I have a quick action which invoke a lighting component.It calls the apex class from there based on user input.The issue is button is working for all the user , but once the user input Yes/No, it closes without any action for non-admin users.It works perfectly if the user is admin.Can anybody help me to understand the issue?

JS Controller
    ({

    yesResponse : function(component, event, helper) {

        var action = component.get("c.recallApproval");

        action.setParams({
          "currId": component.get("v.recordId"),
        });
        //alert(action);
        action.setCallback(this, function(response) {
            var state = response.getState();
            console.log('state '+state);
            if(state == "SUCCESS" && component.isValid()){

               var toastEvent = $A.get("e.force:showToast");
                toastEvent.setParams({
                    "title": "Success!",
                    "message": "Success",
                    "type": "success"
                });
                toastEvent.fire();
            }else{
              console.error("fail:" + response.getError()[0].message);
            }
            $A.get('e.force:refreshView').fire();
            $A.get("e.force:closeQuickAction").fire();
          });
          $A.enqueueAction(action);

    },

    noResponse : function(component, event, helper) {

        alert('Deactivation Cancelled');
        $A.get("e.force:closeQuickAction").fire();
    }
})

apex class
-----------------------------------------------------`
global class approvalRecall
{   @AuraEnabled
    webservice static void recallApproval(id currId)

    {   system.debug('approvalRecall class invoked..');
        List <Costing_Sheet_Item__c> relProd=[Select id,Costing_Sheet__c,Name from Costing_Sheet_Item__c where Costing_Sheet__c=:currId];
        List <Costing_Sheet__c> records=[Select id,Active__c,Approval_Status__c from Costing_Sheet__c where id=:currId];

     for(Costing_Sheet__c sp : records){
         //system.debug('relProd= '+relProd);
         sp.Active__c=False;
         sp.Approval_Status__c='Deactivated';

         system.debug('Approval.isLocked(sp.id)= '+Approval.isLocked(sp.id));
         if(Approval.isLocked(sp.id)){

             Approval.unlock(sp.id);

         }
     }
     update records;
     delete relProd;
    }
}

redpajama_set_name: RedPajamaStackExchange


text:
Vienna State Opera: Ioan Holender Farewell Concert
Soloists; Chorus and Orchestra of the Vienna State Opera (DG DVD 0014913-09)
In order to celebrate his retirement after nineteen years as general director of the Vienna State Opera, Ioan Holender organized the glittering parade of star singers and conductors preserved on these highly entertaining DVDs. Opera is a very serious business in Vienna, and their opera company is among the finest in the world, offering a diverse palette of works over a long, ten-month season from September through June. Along with standard repertoire, Vienna has long been known for offering titles from hidden corners of the repertoire and also for the ambition to mount the grandest works on a massive, expensive scale. Sprawling epics such as Rossini's Guillaume Tell, Wagner's Rienzi, and Verdi's most complete, five-act Don Carlos have been offered in ground-breaking productions that would terrify accountants at the Metropolitan Opera and other leading venues. And such excess has not been restricted to repertoire: Vienna fields an international roster of the finest singers and conductors. Alongside itinerant celebrities who may appear in one or two productions annually, the house also cultivates a strong cadre of 'house singers' who participate in many productions per season – often or exclusively in leading roles. Therefore, it comes as no surprise that the programming for this celebratory gala was chosen to represent all of the aforementioned strengths while recalling most of the forty-or-so new productions offered during Holender's regime.
Apparently, the full concert was slightly abridged for release on DVD, but even still, the running time comes in at three hours, forty-five minutes – an exceptionally generous program filled with lots of exciting singing. Conductor Zubin Mehta started the evening with a somewhat stolid account of the overture to Wagner's Rienzi, after which, Holender himself made a few opening remarks before ceding the stage to the singers. At sixty-nine years old, Plácido Domingo still has the ability to amaze, and once again proved his superhuman vocal durability by traversing a somewhat effortful 'Winterstürme' from Die Walküre. Under Antonio Pappano's sensitive leadership, Domingo celebrated almost twenty years since his debut as Siegmund in 1992, eliciting enthusiastic ovations from his appreciative audience. Bulgarian mezzo-soprano Nadia Krasteva followed with a strongly vocalized but blandly acted 'Stride la vampa' from Verdi's Il Trovatore – always an awkward choice for the concert platform with its lack of scenic and orchestral context. The sextet from Offenbach's Les contes d'Hoffmann (raggedly sung) provided a sonic backdrop for the first multi-media event of the program. Huge screens on the wall behind the orchestra were employed to show scenes from the opera when the production was new. This scheme was used several times throughout the evening, though there seemed to be no particular rationale for which productions were included on video.
General Music Director-designate Franz Welser-Möst made the first of many appearances by conducting two selections from Mozart's Cosi fan tutte. Tenor Michael Schade offered a beautifully phrased 'Un'aura amorosa', and soprano Barbara Frittoli (looking a bit uncomfortable) joined with effervescent mezzo-soprano Angelika Kirchslager in the animated duet 'Sorella, cosa dici?' Frittoli returned later in the program for more Mozart with a poised rendition of the Contessa's difficult 'Dove sono' from Le nozze di Figaro. Schade and Kirchschlager also returned with 'So kommen Sie!' – the charming scene for Valencienne and Camille from Lehár's Die lustige Witwe. Tenor Ramón Vargas made an uncharacteristically bland showing in 'Amor ti vieta' from Giordano's Fedora, showing why light lyric voices are generally ill-suited to verismo. Fortunately, he made a much stronger impression later in the evening with Romeo's 'Ah! leve-toi, soleil!' from Gounod's Shakespearian chestnut, showing excellent breath control but slightly stiff phrasing.
Baritone Boaz Daniel offered the first real misfire of the evening with an out of tune, unstylish rendition of Hérode's 'Vision fugitive' from Massenet's gorgeous Hérodiade. This was one of three selections that might better have been left on the cutting room floor, particularly if other more worthy items were omitted during the editing process prior to commercial release. Tenor Saimir Pirgu's shouted, amateurish rendition of Rinuccio's ardent 'Avete torto! … Firenze e come un albero fiorito' from Puccini's Gianni Schicchi should also have been cut. Finally, soprano Stefania Bonfadelli offered the poorest showing on the DVD with her charmless 'O luce di quest'anima' from Donizetti's Linda di Chamounix. After four minutes of wrong notes, poor intonation, sliding through smudged coloratura, and lunging for high notes, I was mystified to see the audience applauding enthusiastically. Bonfadelli's vocal technique was nearly nonexistent: it was a sad display from a once promising young singer.
At the other end of the spectrum, Soile Isokoski brought supreme control to Agathe's aria 'Und ob die Wolke' from Weber's Der Freischütz. Under the serene direction of Peter Schneider, the orchestra collaborated with Isokoski to take us into Weber's proto-Romantic world, producing one of the most satisfying highlights of the concert. Similarly transporting, Waltraud Meier – looking stunning – offered her well-known intensity in familiar repertoire: Isolde's 'Mild und leise'. This showstopper was followed by another, when Leo Nucci arrived to sing Monforte's 'In braccio alle dovizie' from Verdi's I vespri siciliani. At 68, Nucci retains the amazing intensity of delivery and knowledge of Verdian style that have made him into something of an icon. Rounding out the first DVD, Thomas Quasthoff offered a touching rendition of a rarity: Morosus's 'Wie schön is doch die Musik' from Strauss's Die Schweigsame Frau. Under Fabio Luisi's splendid direction, Thomas Hampson reprised his Guillaume Tell with a dignified 'Sois immobile', and finally, Diana Damrau gave a hint of her future in the bel canto repertoire with the final aria/cabaletta for Amina from Bellini's La Sonnambula. The aria – 'Ah! non credea mirarti' was impressive for its poise and breath control. But while she caught the general spirit of the effervescent cabaletta, Damrau rather overdid the fireworks with some ungainly ornaments, slightly dampening the overall impression.
The second DVD offers almost non-stop enjoyment, beginning with a thrilling finale from Strauss's Die Frau ohne Schatten. Welser-Möst worked his way through the dense orchestration while accompanying a stunning quartet of singers including Falk Struckmann, Johan Botha, Deborah Polaski, and a radiant Adrienne Pieczonka. Polaski seemed unusually muted: I'm not sure I heard any of her vocal lines cutting through the acoustical haze. The other three were fantastic however, with Pieczonka particularly moving as she soared high above the orchestra as the Kaiserin. A new recording of this demanding opera featuring Struckmann, Botha, and Pieczonka would be very welcome, indeed. Botha returned later in the program with a handsomely sung, emotionally resonant rendition of Lohengrin's 'In fernem Land'. Pieczonka also made a second appearance, collaborating with soprano Genia Kühmeier in 'Er ist der Richtige' from Strauss's Arabella. Both sopranos offered sterling vocalism, with Pieczonka particularly impressive by virtue of her gleaming tone, patrician phrasing and presence, and remarkable feeling for the texts. Peter Seiffert and Petra Maria Schnitzer sang a five-minute fragment from the second act of Tristan und Isolde without much subtlety, and were followed by Ferruccio Furlanetto's shattering traversal of Philippe's soliloquy from Verdi's Don Carlos, 'Elle ne m'aime pas!'
Soprano Angela Denoke and tenor Stephen Gould reprised their roles in Korngold's fascinating Die tote Stadt by highlighting the opera's most famous moment of lyricism with 'Glück, das mir verblieb'. Denoke's crystalline clarity touchingly illuminated Marietta's vulnerability and contrasted effectively with Gould's blunt, slightly stentorian Paul. Up-and-coming superstar tenor Piotr Beczala appeared twice on the second half of the concert – much to the delight of the audience. First, he offered an excellent, heartfelt 'Pourquoi me réveiller' from Massenet's wrenching Werther, only showing the slightest strain on high, but displaying masterful vocal technique. His second selection was a soaring, lyrical 'Salut! Demeure chaste et pure' from Gounod's Faust. In both instances, his singing was robust, rich-toned, and beautifully phrased, often calling to mind the great Nicolai Gedda. Bulgarian soprano Krassimira Stoyanova looked stunning and used her poised, silvery voice to make an indelible impression with Anna's seldom heard 'Se come voi piccina io fossi' from Puccini's Le villi. Conductor Simone Young coaxed the maximum poetry from the orchestra, both here and in Botha's ensuing aria from Lohengrin. These moments were magical.
Appearing in a bright fuchsia gown, Anna Netrebko stormed the stage and then thoroughly captivated the audience with 'Je marche sur tous les chemins… Obéissons quand leur voix appelle' from Massenet's Manon. This terrifically difficult scene is a daring choice for the concert stage, since it requires a great deal of vocal 'acting' and contains plenty of coloratura roulades, trills, and high notes. Playing somewhat against type, Netrebko triumphed with agility to spare, excellent, clean execution of the passagework, decent trills, and spot-on high D's. She had the audience in the palm of her hand throughout, and they showered her with the loudest ovations of the evening. Following Netrebko's raw charisma, soprano Natalie Dessay appeared – rather meekly – to sing Marie's aria/cabaletta from Donizetti's La Fille du regiment: 'C'en est donc fait… Salut a la France!' The aria was breathtaking in its control and depth of emotion – particularly given Marco Armiliato's refined conducting, but the cabaletta found Dessay gasping for breaths, physically flailing to spit out the words and phrases, and lunging at high notes. Ending with one of the most effortful high E-flats in recent memory, it became crystal clear why this intelligent soprano has recently chosen to move into more lyric, less high-lying repertoire. Rounding out the program, soprano Violeta Urmana soared through an impressive 'Pace, pace, mio dio!' from Verdi's La forza del destino, baritone Simon Keenlyside thrilled in an introverted reading of Macbeth's scena 'Perfidi! All'Anglo contro me v'unite… Pieta, rispetto, onore', and baritone (!) Adrian Eröd triumphed with a bleeding chunk of Loge's music from Das Rheingold.
After a few closing remarks from Holender, Leo Nucci joined a host of other singers as Falstaff in the rollicking fugue finale from Verdi's opera, 'Tottu nel mondo e burla'. After a long evening of mostly spectacular singing, this closing ensemble – 'Everything in the world is a joke' - provided a fitting conclusion, leaving the audience with hints of philosophy and comedy. Sincere thanks are due to Deutsche Grammophon for undertaking the expense and effort of recording such a grand event and making it available to an international audience. Lovers of great singing need not hesitate: this is a fine addition to any operatic library.
By David Laviska


July 11, 2016 (EIRNS)—The trans-Atlantic press is full of coverage of the Deutsche Bank and Italian banking crises, underscoring the importance of Lyndon LaRouche's intervention, calling for a one-time bailout of Germany to prevent chaos in Europe, but based on the need for major changes in the banking system, to push credits into the real economy and shut down the derivatives and other speculative bubbles.
Russia Today picks up on David Folkerts-Landau's interview with Welt am Sonntag, in which the chief economist of Deutsche Bank (DB) called for an emergency 150-billion-euro bailout, and also cites statements from Lorenzo Bini Smaghi, chairman of Société Générale, warning that the Italian banking crisis can spread to all of the EU. DB stocks are down 48 percent over the past 12 months, Société Générale stock is down 63 percent, and the Bloomberg Europe index of 500 banks and financial service firms is down 33 percent, reaching a seven-year low. A timeline in the Bloomberg article is titled "The Epic Collapse of Deutsche Bank," which makes clear that the European banking system is now, immediately, on the edge of a crash.
RT also picks up on a recent, signed op-ed by George Soros, declaring that the collapse of the EU has become "almost inevitable" since the Brexit vote.
"The catastrophic scenario that many feared has materialized, making the disintegration of the EU practically irreversible," he wrote for Project Syndicate (which he heavily funds). He added that the financial collapse in the UK following Brexit is the worst in three decades. "The very survival of the European project" is on the line in the negotiations of how Brexit will occur. RT coverage noted that Marine LePen met with French President Hollande and pressed for a referendum on "Frexit," but was rebuffed.
A Wall Street Journal chart cited by The Street shows the bank-to-bank connections of DB. DB is leveraged more than 40:1, far worse than Lehman's 31:1 at the time of its collapse; and its current derivatives portfolio is $72.8 trillion, 13 percent of all global outstanding derivatives.
New Europe online headlines, "Why Deutsche Bank is the most dangerous bank in the world," asking what the price would be for the German government to bail it out, versus the consequences of letting it blow out with systemic implications.
Bloomberg also warned that the London real estate market is crashing and this is another consequence of Brexit. Standard Life Investments announced that as of today, it will suspend its UK Real Estate fund, to ward off investors demanding their money back. This is already triggering contagion, with several other big real estate investors announcing similar freezes on clients' funds, and others announcing simply that they are pulling out of existing deals for prime London real estate projects.
Taken together, the conditions of Italy, Deutsche Bank, and the London real estate market are more than sufficient to blow up the entire trans-Atlantic financial sector. It is precisely because of this already-onrushing crisis that emergency actions, precisely along the lines demanded by LaRouche, must be enacted immediately.


text:
Prince William Surprises Fans Honouring Princess Diana
"I think I'm still shaking now. I feel very emotional," one fan said about his visit.
By Carly Ledbetter, HuffPost US
Prince William gave well-wishers the surprise of their life when he stopped by a vigil honouring the late Princess Diana's birthday.
The Duke of Cambridge emerged from Kensington Palace on Monday night to say thank you to those gathered to remember his mother who was born on July 1, 1961, the Daily Mail reported.
"William told me he knew we'd been coming here for years and thanked us for what we were doing for his mother," John Loughrey, a 59-year-old admirer, told the outlet. "I think I'm still shaking now. I feel very emotional."
Loughrey said that after shaking William's hand, he asked the prince about the forthcoming Diana memorial that will stand at Kensington Palace.
"Soon, very soon," the prince said. "We just want to make sure it is right. It's important to get it right."
Anwar Hussein via Getty Images
Prince William with Diana, Princess of Wales and Prince Harry on the day he joined Eton in Sept. 1995.
In the past few years, Princes Harry and William have become more candid about dealing with grief in the wake of their mother's unexpected death from a car crash in Paris in 1997.
Harry said that for a long time, he just closed off his emotions, rather than deal with them.
"I can safely say that losing my mum at the age of 12, and therefore shutting down all of my emotions for the last 20 years, has had a quite serious effect on not only my personal life but my work as well," the Duke of Sussex said in a 2017 interview with The Telegraph.
EMPICS Entertainment
William and Harry look at tributes left by the public dedicated to their mother Princess Diana following their visit to The White Garden, in Kensington Palace on Aug. 30, 2017.
He added: "My way of dealing with it was sticking my head in the sand, refusing to ever think about my mum because why would that help? It's only going to make you sad. It's not going to bring her back."
Later, he turned to counseling and boxing at the age of 28 to begin working through his struggles.
"The experience I have had is that once you start talking about it, you realize that actually you're part of quite a big club," Harry said.
This Advice From Princess Diana May Have Helped Her Sons Find Love
There's A Newfoundlander In The Royal Family. NBD.
Prince Harry Is Taking Up Another One Of His Mother's Beloved Causes
Carly Ledbetter Huffington Post Entertainment Writer
MORE: Royal Family Prince William Princess Diana