Lessons from CanSpotASF: Moving towards risk-based African Swine Fever surveillance with rule-out testing in Western Canada.

African swine fewer (ASF) is a serious disease present in Africa, Eurasia, and the Caribbean but not in continental North America. CanSpotASF describes the ASF surveillance in Canada as a phased in approach. The first enhancement to the passive surveillance was the risk-based early detection testing (rule-out testing) where eligible cases were tested for ASF virus (ASFv). The objective was to describe how the eligibility criteria were applied to cases in western Canada. In particular, to assess if cases tested for ASFv had eligible conditions and if pathology cases with eligible conditions were tested for ASFv based on the data collated by Canada West Swine Health Intelligence Network (CWSHIN) from British Columbia, Alberta, Saskatchewan, and Manitoba. The study period was August 2020 to December 2022 and the data included two study laboratories. We found that over 90% of cases tested for ASFv had eligible conditions as defined in CanSpotASF. The eligibility criteria were applied at three stages of the disease investigation process: 1) the clinical presentation in the herd; 2) at the initial laboratory assessment; and 3) the final pathology diagnosis. At the two study laboratories the proportion of all submitted cases (culture, serology, PCR, pathology) tested for ASFv was very low 1%. However, in the pathology cases specifically targeted in CanSpotASF, and the proportion of tested cases was 12%. In addition, for eligible pathology cases (eligible diagnosis or test) the proportion tested was higher 15%. These results indicated that CanSpotASF targeted herds with submissions for pathological examination and to some degree eligible conditions which would be herds with health issues (known or unknown). We interpret this as a first step towards risk-based surveillance with health as the defining factor.

A SACS deletion variant in Great Pyrenees dogs causes autosomal recessive neuronal degeneration.

ARSACS (autosomal recessive spastic ataxia of Charlevoix-Saguenay) is a human neurological disorder characterized by progressive cerebellar ataxia and peripheral neuropathy. A recently recognized disorder in Great Pyrenees dogs is similarly characterized by widespread central nervous system degeneration leading to progressive cerebellar ataxia and spasticity, combined with peripheral neuropathy. Onset of clinical signs occurred in puppies as young as 4 months of age, with slow progression over several years. A multi-generation pedigree suggested an autosomal recessive mode of inheritance. Histopathology revealed consistent cerebellar Purkinje cell degeneration, neuronal degeneration in brainstem nuclei, widespread spinal cord white matter degeneration, ganglion cell degeneration, inappropriately thin myelin sheaths or fully demyelinated peripheral nerve fibers, and normal or only mild patterns of denervation atrophy in skeletal muscles. Genome-wide single nucleotide polymorphism (SNP) genotype data was collected from 6 cases and 26 controls, where homozygosity mapping identified a 3.3 Mb region on CFA25 in which all cases were homozygous and all controls were either heterozygous or homozygous for alternate haplotypes. This region tagged the SACS gene where variants are known to cause ARSACS. Sanger sequencing of SACS in affected dogs identified a 4 bp deletion that causes a frame shift and truncates 343 amino acids from the C terminus of the encoded sacsin protein (p.Val4244AlafsTer32). Our clinical and histopathological descriptions of this canine disorder contribute to the description of human ARSACS and represents the first naturally occurring large animal model of this disorder.

Computation Offloading and User-Clustering Game in Multi-Channel Cellular Networks for Mobile Edge Computing.

Mobile devices may use mobile edge computing to improve energy efficiency and responsiveness by offloading computation tasks to edge servers. However, the transmissions of mobile devices may result in interference that decreases the upload rate and prolongs transmission delay. Clustering has been shown as an effective approach to improve the transmission efficiency for dense devices, but there is no distributed algorithm for the optimization of clustering and computation offloading. In this work, we study the optimization problem of computation offloading to minimize the energy consumption of mobile devices in mobile edge computing by adaptively clustering devices to improve the transmission efficiency. To address the optimization problem in a distributed manner, the decision problem of clustering and computation offloading for mobile devices is formulated as a potential game. We introduce the construction of the potential game and show the existence of Nash equilibrium in the game with a finite enhancement ability. Then, we propose a distributed algorithm of clustering and computation offloading based on game theory. We conducted a simulation to evaluate the proposed algorithm. The numerical results from our simulation show that our algorithm can improve offloading efficiency for mobile devices in mobile edge computing by improving transmission efficiency. By offloading more tasks to edge servers, both the energy efficiency of mobile devices and the responsiveness of computation-intensive applications can be improved simultaneously.

Bioinformatic analysis of dysregulated circular RNAs in pediatric pulmonary hypertension linked congenital heart disease.

Background: Circular RNAs (circRNAs) may play important roles in the progression of pulmonary arterial hypertension. However, the potential roles they play in childhood pulmonary arterial hypertension associated congenital heart disease (CHD) progression remains unclear. Methods: Thirteen human plasma samples including eight from pulmonary arterial hypertension secondary to CHD patients and five from a control group were analyzed using the Arraystar Human circRNA array. The relative expression levels of five differentially expressed circRNAs in pulmonary arterial hypertension were detected using real-time polymerase chain reaction (PCR) analysis. In parallel, these levels were also taken on control samples from 32 CHD patients. We used miRanda and TargetScan software packages to predict potential microRNA (miRNA)targets, which were then combined into a circRNA-miRNA-messenger RNA (mRNA) network. Results: Twenty-seven circRNAs (three upregulated and 24 downregulated) were differentially expressed between the pulmonary arterial hypertension and control groups. Compared to control group levels, circ_003416 expression in the pulmonary arterial hypertension group was significantly downregulated, while circ_005372 expression, in contrast, was significantly upregulated. The differential expression of these circRNAs was mainly linked to variation in levels of oxidative phosphorylation and tight junction signaling. Conclusions: We identified one overexpressed and one underexpressed circRNA in plasma samples from children with CHD associated pulmonary arterial hypertension. Bioinformatic analysis indicated these dysregulated circRNAs might be associated with the occurrence and regulation of pulmonary arterial hypertension.

Effects of YM155 on the proliferation and apoptosis of pulmonary artery smooth muscle cells in a rat model of high pulmonary blood flow-induced pulmonary arterial hypertension.

INTRODUCTION: Proliferation and apoptosis of pulmonary artery smooth muscle cells (PASMCs) play an important role in the occurrence and development of pulmonary arterial hypertension (PAH). The purpose of this study was to investigate the effects of survivin inhibitor YM155 on the proliferation and apoptosis of PASMCs in rats with PAH induced by high pulmonary blood flow. METHODS: Thirty male Sprague-Dawley (SD) rats were randomly divided into control, model, and YM155 intervention groups. A rat model of PAH induced by high pulmonary blood flow was established, and it was confirmed by assessments of right-ventricular pressure (RVP) and right ventricular hypertrophy index (RVHI). Immunohistochemical staining and western blot analysis were used to detect the expression of survivin, and the proliferation and apoptosis of PASMCs. Lastly, the effects of in vivo treatment of YM155 were tested. RESULTS: The increased expression of survivin mRNA and protein were observed in the model group, accompanied by pulmonary arteriolar wall thickening, lumen stenosis, and perivascular inflammatory cell infiltration. Elevated expression of survivin and pulmonary vascular remodeling were significantly mitigated after YM155 treatment. Specifically, the YM155 intervention group had a significantly lower PASMC proliferation rate and a higher PASMC apoptotic rate. CONCLUSION: YM155 suppressed PASMC proliferation and promoted PASMC apoptosis by inhibiting survivin expression and thereby reducing pulmonary vascular remodeling in high pulmonary blood flow-induced PAH in vivo.

Risk Factors for Resistance to Intravenous Immunoglobulin Treatment and Coronary Artery Abnormalities in a Chinese Pediatric Population With Kawasaki Disease: A Retrospective Cohort Study.

Background: The factors predicting high-risk Kawasaki disease (KD) remain unclear. Therefore, we aimed to determine the risk factors for resistance to intravenous immunoglobulin (IVIG) treatment and coronary artery aneurysm (CAA) development in a Chinese pediatric population with high-risk KD. Methods: We compared the performances of 11 scoring systems that have been reported to predict IVIG resistance among patients with KD hospitalized from January 2013 through August 2021. Patients were risk-stratified based on the optimal scoring system. The association of baseline characteristics with IVIG treatment resistance and CAA development was investigated within the high-risk group of KD. Results: In total, 346 pediatric patients with KD were included, of whom 63 (18.2%) presented with IVIG resistance. The Kobayashi score and five Chinese scoring system scores (Tang et al., Yang et al., Lan et al., Liping et al., and Wu et al.) were significantly higher in the IVIG non-responsive KD group than in the IVIG responsive KD group, and the results of the receiver operating characteristic (ROC) curves analysis were observed to be highest in the Xie Liping scoring system for IVIG resistance (area under the curve, 0.650). Especially, 87 (25.1%) patients comprised the high-risk KD group based on this optimal scoring system (≥5 points). IVIG resistance was significantly associated with the total bilirubin-to-albumin ratio (B/A ratio) [odds ratio, 7.427; 95% confidence interval (CI): 1.022-53.951]. The area under the ROC was 0.703 (95% CI: 0.586-0.821), and the cutoff point was 0.383, which indicated a sensitivity and specificity for predicting treatment resistance of 58% and 80%, respectively. The serum albumin level (odds ratio, 1.401; 95% CI: 1.049-1.869) and Z score of the left main coronary artery (odds ratio, 9.023; 95% CI: 1.070-76.112) were independent predictors of CAA development. Conclusions: In the Chinese pediatric population with KD, the Xie Liping scoring system is the most appropriate method for identifying high-risk patients, and IVIG resistance could be predicted based on the B/A ratio. Serum albumin level and Z score of the left main coronary artery at baseline were warning indicators for CAA development. More intensified or adjunctive therapies and close follow-up should be considered for high-risk patients with these risk factors.

Phylogenetic analysis of porcine circovirus 3 circulating in Canadian pigs.

INTRODUCTION: Porcine circovirus 3 (PCV3) has been detected in pigs worldwide and associated with several clinical signs. METHODS: To investigate the genetic diversity of PCV3 strains circulating in Canada, 44 PCV3 positive samples from Saskatchewan (2/44), Manitoba (2/44), Quebec (4/44), Alberta (11/44) and Ontario (25/44) submitted to diagnostic laboratories in Canada between 2019 and 2021 were sequenced and analyzed. RESULTS: Phylogenetic analysis of capsid genes showed that all of the 44 Canadian strains classified into PCV3a and segregated into seven lineages with common amino acid changes observed at A24V, R27K, N56D, T77S, Q98R, L150I (F) and R168K positions. CONCLUSION: Future studies are required to determine whether the polymorphisms in capsid proteins, as revealed in this study, could be associated with differences in the pathogenicity or antigenicity of PCV3 strains. This is the first phylogenetic analysis of PCV3 strains among different provinces in Canada.

Streptococcus equi subsp. zooepidemicus infection of pigs leads to shedding in faeces and a carrier state.

In 2019, Streptococcus equi subsp. zooepidemicus was recognized as an emerging pathogen of swine, associated with sudden deaths, increased abortion rates and septicaemia. Limited data are available regarding this disease in pigs. The objectives of this study were to clarify clinical progression, pathogen shedding, transmission, gross and microscopic lesions following infection in pigs. Six weeks old pigs were inoculated with either S. zooepidemicus sequence type 194 (inoculated, n = 6) or sham inoculated with sterile culture broth (sentinels, n = 4). Animals were housed in the same room, in two pens 2 m apart. Pigs were monitored twice daily for clinical signs, and rectal, nasal and oral swabs were collected once daily. A full necropsy was performed if welfare was a concern or at 5 days post-inoculation (dpi). All sentinels remained disease free and their samples tested negative for the pathogen of interest. All inoculated pigs developed fever within 8 h of inoculation, and severe disease was observed after 2 dpi. A total of 4/6 inoculated pigs developed clinical signs that compromised animal welfare and were euthanized. Nasal swabs (15/23), followed by rectal swabs (9/23) yield the highest number of positive ante-mortem samples. Clinically healthy, inoculated pigs had detectable levels of S. zooepidemicus in rectal and nasal swabs. Reactive submandibular lymph nodes, kidney petechiae and splenomegaly were found in six of six inoculated pigs. These data suggest that subclinically infected pigs may spread the pathogen through nasal secretions and faeces. Direct contact seems to be required for transmission.

Expression and clinical significance of circular RNA hsa_circ_0003416 in pediatric pulmonary arterial hypertension associated with congenital heart disease.

BACKGROUND: Circular RNAs (circRNAs) have been found to be involved in the development of pulmonary arterial hypertension (PAH). However, their diagnostic value in pediatric PAH remains unclear. This study aimed to examine the characteristic expression of the circRNA hsa_circ_0003416 in the plasma of children with PAH caused by congenital heart disease (CHD); the potential of hsa_circ_0003416 as a diagnostic biomarker was also investigated. METHODS: The plasma expression levels of hsa_circ_0003416 were determined via quantitative reverse transcription-polymerase chain reaction in 50 CHD patients, 50 PAH patients, and 20 healthy subjects; the associations between hsa_circ_0003416 levels and clinical data were analyzed thereafter. Receiver operating characteristic curves were employed to determine the diagnostic capacity of this circRNA. RESULTS: Expression levels of hsa_circ_0003416 in plasma were lower in the PAH-CHD group than in the CHD and healthy control groups (p = 0.009 vs. healthy control group, p = 0.026 vs. CHD group). Moreover, hsa_circ_0003416 was found to be negatively associated with B-type natriuretic peptide (r = -0.342, p = 0.013). In addition, the area under the curve of hsa_circ_0003416 levels in plasma was 0.721 (95% confidence intervals = 0.585-0.857, p = 0.004), suggesting that it has a promising diagnostic value. CONCLUSIONS: Overall, hsa_circ_0003416 was found to be significantly downregulated in children with PAH-CHD and to be potent as a biomarker for PAH-CHD diagnosis.

Domestic pigs experimentally infected with Mycobacterium bovis and Mycobacterium tuberculosis exhibit different disease outcomes.

The domestic pig shares many similarities with humans in anatomy, physiology, and immunology. As such it is an attractive animal model to study human tuberculosis (TB). In this study, we examined disease outcome in pigs challenged via two different routes with either the human TB bacillus Mycobacterium tuberculosis Erdman (M. tb) or bovine TB bacillus M. bovis AF2122/97 in head-to-head comparisons. Pigs challenged intravenously with M. bovis exhibited greater morbidity and rapid onset of mortality, higher bacterial burden and tissue necrosis compared to pigs challenged similarly with M. tb. Concordantly, pigs challenged with aerosolized M. bovis exhibited reduced weight gain and more severe pathology than pigs challenged similarly with M. tb. Specifically, M. bovis challenged pigs presented a spectrum of granulomatous lung lesions similar to that in human TB. In contrast, pigs challenged with M. tb presented mostly early-stage granulomas. Irrespective of challenge dose and pathology however, peripheral IFN-γ responses were similar in both M. bovis and M. tb aerosol challenged pigs. Although M. bovis appears to be more virulent than M. tb, both can be used to model different facets of human TB in pigs, depending on whether one seeks to recapitulate active or latent forms of the disease.

Metabolic Reprogramming of the Right Ventricle and Pulmonary Arteries in a Flow-Associated Pulmonary Arterial Hypertension Rat Model.

Pulmonary arterial hypertension (PAH) is a complex devastating disease relevant to remarkable metabolic dysregulation. Although various research studies on PAH from a metabolic perspective have been emerging, pathogenesis of PAH varies in different categories. Research on metabolic reprogramming in flow-associated PAH remains insufficient. An untargeted metabolomic profiling platform was used to evaluate the metabolic profile of pulmonary arteries (PAs) as well as the right ventricle (RV) in a flow-associated PAH rat model in the present work. A total of 79 PAs and 128 RV metabolites were significantly altered in PAH rats, among which 39 metabolites were assessed as shared dysregulated metabolites in PAs and the RV. Pathway analysis elucidated that, in PAs of PAH rats, pathways of phenylalanine, tyrosine, and tryptophan biosynthesis and linoleic acid metabolism were significantly altered, while in the RV, arginine biosynthesis and linoleic acid metabolism were altered dramatically. Further integrated analysis of shared dysregulated PA and RV metabolites demonstrated that the linoleic acid metabolism and the arachidonic acid (AA) metabolism were the key pathways involved in the pathogenesis of flow-associated PAH. Results obtained from the present work indicate that the PAH pathogenesis could be mediated by widespread metabolic reprogramming. In particular, the dysregulation of AA metabolism may considerably contribute to the development of high blood flow-associated PAH.

Partial reproduction of ear-tip necrosis suggests an infectious, initially bacterial aetiology.

Swine ear-tip necrosis (ETN) is a disease of global presence and unclear aetiology. Little evidence is available regarding the nature of this disease. The aim of this work was to investigate if ETN is an infectious disease that could be replicated using a lesion macerate inoculum. A source farm with a history of ear-tip necrosis was identified and five weeks-old pigs (n = 12) from this farm were housed under controlled conditions and intradermally inoculated with ETN lesion macerates (right ear, n = 10) or sterile inoculum (left ear, n = 10). Two pigs were not inoculated, serving as sentinels. All animals were clinically monitored daily during 21 days, and a ETN ear score was used to follow disease progression. Anaerobic (n = 2) and aerobic (n = 2) overnight cultures, as well as raw aliquots of the lesion macerate inoculum (n = 2) and control inoculum (n = 2) were submitted for metagenomic sequencing. All inoculated ears developed lesions suggestive of early ETN, but none progressed to result in loss of the ear pinna. All completely resolved 21 days post-inoculation. Post-mortem investigation revealed areas of fibrosis, characterized by a granulomatous response in the inoculated ears (5/10) and in 1/10 control ears. Metagenomic analysis identified the presence of previously suggested bacterial etiological agents, but no relevant viral, fungal or protozoan agents in the inoculum. ETN etiology remains unclear, but an infectious cause and bacterial agents are suggested to be at least partially implicated in disease pathogenesis. Virus and fungi do not seem to significantly contribute to disease.

Development and evaluation of a new method to combine clinical impression survey data with existing laboratory data for veterinary syndromic surveillance with the Canada West Swine Health Intelligence Network (CWSHIN).

The Canada West Swine Health Intelligence Network (CWSHIN) is a surveillance system imbedded in an intelligence network. It has been conducting syndromic surveillance in the four western provinces of Canada since 2012. The quarterly activities include repeated clinical impression surveys, compilation of laboratory data, discussion of trends with an expert group (practitioners, laboratory diagnosticians) and swine health reports for producers and swine practitioners. However, due to the lack of standardized population identifiers across data sources usual methods of combining data could not be applied and the collated data were not being fully utilized and analysed. Therefore in 2019, CWSHIN underwent a substantial review resulting in the "Next Generation CWSHIN". The objectives of this study were to develop and evaluate a new data merging method to combine CWSHIN's clinical impression survey and laboratory data; and to provide examples of analyses and modeling based on these data. The data for analysis were restricted to repeated clinical impression surveys (2019-2020) from veterinary practitioners and laboratory diagnostic data (2016-2020). Merging surveillance data from existing sources can be challenging. Therefore, as an alternative to merge data using a hierarchy of population identifiers, we developed a Disease Map to link surveillance data from all our data-sources. The resulting Data Repository allowed monitoring of temporal trends of syndromes, clinical diseases, and laboratory identified organisms, but it cannot provide estimates of disease occurrence. Two main reasons were the lack of denominators and using existing data on routine diagnostic tests. Therefore, discussion in the expert group (veterinary practitioners, laboratory diagnosticians, swine health experts) was critical to the system's success. Based on repeated clinical impression surveys a stochastic scenario tree model for freedom from Foot and Mouth Disease (CWSHIN Blister model) was also developed. In conclusion, the method to link existing data systems from multiple divergent sources by means of a Disease Map improved CWSHIN's veterinary syndromic surveillance. Together the Data Repository and Disease map provided flexibility to monitor temporal trends, define populations and diseases, and allow analysis. However, it is critical that the surveillance is coupled with a good intelligence network that can help interpret the results and disseminate knowledge to veterinarians and producers.

Clinical Characteristics and Factors Associated With Hypertension in 205 Hospitalized Children: A Single-Center Study in Southwest China.

Objective: The aim of this study was to investigate the clinical characteristics and factors associated with pediatric hypertension and target organ damage (TOD). Methods: We retrospectively reviewed clinical data from 205 children with hypertension treated in our hospital from 2007 to 2018. The patients were classified based on the type of hypertension (primary, secondary) and presence of TOD (heart, brain, retina). Logistic regression analysis was performed to identify the factors independently associated with hypertension and TOD. Results: There were 107 males, 97 females, and one intersex in this study, with an age range of 0.1-17.9 years. Majority of cases (177, 86.3%) had secondary hypertension, while 13.7% had primary hypertension. The most frequent cause of secondary hypertension was renal disease (59.32%). Elevated serum creatinine level (odds ratio [OR] = 7.22, 95% confidence interval [95% CI] = 1.6-32.62, P = 0.01), blood urea nitrogen (OR = 6.33, 95% CI = 1.81-22.19, P = 0.004), serum uric acid level (OR = 3.66, 95% CI = 1.20-11.22, P = 0.023), and albuminuria (OR = 3.72, 95% CI = 1.50-9.26, P = 0.005) were independently associated with secondary hypertension. Elevated serum uric acid and blood urea nitrogen levels were associated with left ventricular hypertrophy (OR = 6.638, 95% CI = 1.349-32.657, P = 0.02) and hypertensive encephalopathy (OR = 4.384, 95% CI = 1.148-16.746, P = 0.031), respectively. Triglyceride level correlated with hypertensive retinopathy (P = 0.001). Conclusion: Pediatric hypertension was most often secondary, with renal disease as the leading cause. Elevated levels of serum uric acid, blood urea nitrogen, serum creatinine, and albuminuria may indicate secondary hypertension in childhood. Elevated serum uric acid, blood urea nitrogen, and triglyceride levels were associated with left ventricular hypertrophy, hypertensive encephalopathy, and hypertensive retinopathy, respectively.

Association of LncRNA-GAS5 gene polymorphisms and PBMC LncRNA-GAS5 level with risk of systemic lupus erythematosus in Chinese population.

Growth arrest-specific 5 (GAS5) is a kind of long non-coding RNAs (lncRNAs). Previous studies showed that down-regulation of LncRNA-GAS5 was involved in the development of systemic lupus erythematosus (SLE). However, the regulatory mechanism of down-expressed LncRNA-GAS5 in SLE remains obscure. In this study, we aimed to investigate the association of LncRNA-GAS5 polymorphism with SLE risk. And further explore how LncRNA-GAS5 is involved in the occurrence of SLE. Here, we evaluated the relationship between the risk for the development of SLE and the 5-base pair (AGGCA/-) insertion/deletion (I/D) polymorphism (rs145204276) in the LncRNA-GAS5 promoter region. A custom 36-Plex SNPscan kit was used for genotyping the LncRNA-GAS5 polymorphisms. The LncRNA-GAS5 and miR-21 target prediction was performed using bioinformatics software. Enzyme-linked immunosorbent assay (ELISA) and quantitative real-time PCR (qRT-PCR) were performed to assess GAS5 and miR-21 mRNA expression and PTEN protein expression. The results revealed that rs145204276 resulted in a decreased risk of SLE (DD genotypes vs II genotypes: adjusted OR = 0.538, 95% CI, 0.30-0.97, P = .039; ID genotypes vs II genotypes: adjusted OR = 0.641, 95% CI, 0.46-0.89, P = .007; ID/DD genotypes vs II genotypes: adjusted OR = 0.621, 95% CI, 0.46-0.84, P = .002; D alleles vs I alleles: adjusted OR = 0.680, 95% CI, 0.53-0.87, P = .002). A reduced incidence of renal disorders in SLE was found to be related to ID/DD genotypes and D alleles (ID/DD genotypes vs II genotypes: OR = 0.57, 95% CI, 0.36-0.92, P = .020; D alleles vs I alleles: OR = 0.63, 95% CI, 0.43-0.93, P = .019). However, no significant association of rs2235095, rs6790, rs2067079 and rs1951625 polymorphisms with SLE risk was observed (P > .05). Additionally, haplotype analysis showed that a decreased SLE risk resulted from the A-A-C-G-D haplotype (OR = 0.67, 95% CI, 0.49-0.91, P = .010). Also, patients in the SLE group showed a down-regulated expression of LncRNA-GAS5 and PTEN than the healthy volunteers; however, patients with rs145204276 ID/DD genotypes showed up-regulated expression of LncRNA-GAS5 and PTEN compared with patients carrying the II genotype. Furthermore, the miR-21 levels were considerably up-regulated in the SLE group than the healthy volunteers, and patients with rs145204276 ID/DD genotype had lower miR-21 levels than the ones with the II genotype. Thus, we found that the LncRNA-GAS5/miR-21/PTEN signalling pathway was involved in the development of SLE, where LncRNA-GAS5 acted as an miR-21 target, and miR-21 regulated the expression of PTEN. These findings indicated that the rs145204276 ID/DD genotypes in the LncRNA-GAS5 gene promoter region may be protected against SLE by up-regulating the expression of LncRNA-GAS5, which consecutively regulated miR-21 and PTEN levels.

The nasal viromes of cattle on arrival at western Canadian feedlots and their relationship to development of bovine respiratory disease.

Bovine respiratory disease (BRD) has a complex pathogenesis and aetiology, being the costliest disease affecting the cattle industry in North America. In this study, we applied Nanopore-based viral metagenomic sequencing to explore the nasal virome of cattle upon arrival at feedlot and related the findings to the development of BRD. Deep nasal swabs (DNS) from 310 cattle for which BRD outcomes were known (155 cattle developed BRD within 40 days and 155 remained healthy) were included. The most prevalent virus in on-arrival samples was bovine coronavirus (BCV) (45.2%, 140/310), followed by bovine rhinitis virus B (BRBV) (21.9%, 68/310), enterovirus E (EVE) (19.6%, 60/310), bovine parainfluenza virus 3 (BPIV3) (10.3%, 32/310), ungulate tetraparvovirus 1 (UTPV1) (9.7%, 30/310) and influenza D virus (7.1%, 22/310). No relationship was found between BRD development and the number of viruses detected, the presence of any specific individual virus or combination of viruses. Bovine kobuvirus (BKV) was detected in 2.6% of animals (8/310), being the first report of this virus in Canada. Results of this study demonstrate the diversity of viruses in bovine DNS collected upon arrival at feedlot and highlights the need for further research into prediction of BRD development in the context of mixed infections.

TMEM16A Regulates Pulmonary Arterial Smooth Muscle Cells Proliferation via p38MAPK/ERK Pathway in High Pulmonary Blood Flow-Induced Pulmonary Arterial Hypertension.

OBJECTIVE: Pulmonary arterial hypertension (PAH) is a complex disease of the small pulmonary arteries that is mainly characterized by vascular remodeling. It has been demonstrated that excessive proliferation of pulmonary arterial smooth muscle cells (PASMCs) plays a pivotal role in vascular remodeling during PAH. The present study was undertaken to explore the role of TMEM16A in regulating PASMCs proliferation in high pulmonary blood flow-induced PAH. METHODS: Aortocaval shunt surgery was undertaken to establish an animal model. Pulmonary artery pressure and pulmonary vascular structure remodeling (PVSR) were tested. Immunohistochemical staining and Western blot were performed to investigate the expression of TMEM16A. The proliferation of PASMCs was tested by the MTT assay. After treating PASMCs with TMEM16A-siRNA, the expression of proliferating cell nuclear antigen (PCNA), phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK), and phosphorylated extracellular signal-regulated kinase (p-ERK) signaling in PASMCs were tested. RESULTS: PAH and PVSR developed 11 weeks postoperation. Elevated expression of TMEM16A accompanied by high expression of PCNA in pulmonary arteries of the shunt group was observed. The increased proliferation of PASMCs and increased expression of TMEM16A and PCNA, along with activated p-p38MAPK and p-ERK signaling in PASMCs of the shunt group, were all attenuated by siRNA-specific TMEM16A knockdown. CONCLUSION: TMEM16A regulates PASMCs proliferation in high pulmonary blood flow-induced PAH, and the p38MAPK/ERK signaling pathway is probably involved.

Infection With a Novel Rickettsiella Species in Emperor Scorpions (Pandinus imperator).

Rickettsiella infection was diagnosed in 4 adult emperor scorpions (Pandinus imperator) from 2 different collections over a 3-year period. One case had a 2-day history of weakness, failure to lift the tail, or respond to stimulation, with rapid progression to death. The other 3 cases were found dead. There were no gross lesions, but histologically the hemolymphatic vasculature and sinuses, presumed hematopoietic organ, heart, midgut and midgut diverticula, nerves, and skeletal muscle were infiltrated with phagocytic and granular hemocytes with necrosis. Phagocytic hemocytes contained abundant intracellular microorganisms that were Fite's acid-fast-positive, Macchiavello-positive, variably gram-positive or gram-negative, and Grocott's methenamine silver-negative. By transmission electron microscopy, hemocytes contained numerous phagocytic vacuoles with small dense bacterial forms (mean 0.603 × 0.163 µm) interspersed with large bacterial forms (mean 1.265 × 0.505 µm) and few intermediary forms with electron-dense nucleoids and membrane-bound crystalline arrays (average 4.72 µm). Transmission electron microscopy findings were consistent with bacteria of the family Coxiellaceae. Based on sequencing the 16S ribosomal RNA gene, the identity was confirmed as Rickettsiella, and phylogenetic analysis of protein-coding genes gidA, rspA, and sucB genes suggested the emperor scorpion pathogen as a new species. This study identifies a novel Rickettsiella causing infection in emperor scorpions and characterizes the unique pathological findings of this disease. We suggest this organism be provisionally named Rickettsiella scorpionisepticum.

Assessment of Metagenomic Sequencing and qPCR for Detection of Influenza D Virus in Bovine Respiratory Tract Samples.

High throughput sequencing is currently revolutionizing the genomics field and providing new approaches to the detection and characterization of microorganisms. The objective of this study was to assess the detection of influenza D virus (IDV) in bovine respiratory tract samples using two sequencing platforms (MiSeq and Nanopore (GridION)), and species-specific qPCR. An IDV-specific qPCR was performed on 232 samples (116 nasal swabs and 116 tracheal washes) that had been previously subject to virome sequencing using MiSeq. Nanopore sequencing was performed on 19 samples positive for IDV by either MiSeq or qPCR. Nanopore sequence data was analyzed by two bioinformatics methods: What's In My Pot (WIMP, on the EPI2ME platform), and an in-house developed analysis pipeline. The agreement of IDV detection between qPCR and MiSeq was 82.3%, between qPCR and Nanopore was 57.9% (in-house) and 84.2% (WIMP), and between MiSeq and Nanopore was 89.5% (in-house) and 73.7% (WIMP). IDV was detected by MiSeq in 14 of 17 IDV qPCR-positive samples with Cq (cycle quantification) values below 31, despite multiplexing 50 samples for sequencing. When qPCR was regarded as the gold standard, the sensitivity and specificity of MiSeq sequence detection were 28.3% and 98.9%, respectively. We conclude that both MiSeq and Nanopore sequencing are capable of detecting IDV in clinical specimens with a range of Cq values. Sensitivity may be further improved by optimizing sequence data analysis, improving virus enrichment, or reducing the degree of multiplexing.