• The Journal of Korean Physical Therapy
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• 제9권1호
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• pp.59-70
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• 1997

The ability of neurotropic alpha herpesviruses to replicate within synaptically linked neurons has made these pathogens valuable tools for transneuronal analysis. Recent studies suggest that unique gene products expressed by genetically engineered strains of virus may permit the use of multiple strains in complex tracing paradigms. In the present study we have examined the invasiveness of two genetically engineered strains of the swine pathogen known as pseudorabies virus(PRV). The two strains were isogenic with the attenuated Bartha strain of PRV; in one strain a lacZ reporter gene was inserted into the gC locus (PRV-BaBlu; $4.75\times10^8pfu/ml$) contrained a PRV envelope glycoprotein gene that was absent in PRV-BaBlu. Simultaneous or temporally separated sequential injection of $4\mu\ell$ of each strain into the ventral wall of the stomach produced a predictale course of retrograde synaptic infection. The results were as follows: 1. PRV-BaBlu and PRV-D infected the dorsal motor nucleus of vagus nerve(DMV) and paraventricular nucleus(PVN). 2. Invasion and replication of PRV-D occured at a faster rate than the parental strain or PRV-BaBlu. 3. PRV-D was much more virulent than PRV-BaBlu or the parental strain. 4. Co-injection of PRV-D and PRV-BaBlu produced an infection that was more virulent than that produced by the parental strain (PRV-Bartha), 5. Neurons in DMV were permissive to co-infection with PRV-D and PRV-BaBlu when they were injected simultaneously into the same site. 6. Replication of PRV-BaBlu was compromised by prior infection of the same circuit with PRV-D. 7. Prior infection of neurons with PRV-D maked them resistant to infection with PRV-BaBlu.

• 대한한의학회지
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• 제22권2호
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• pp.31-40
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• 2001

This experimental studies was to investigate location of labeled neurons in CNS following injection of pseudorabies virus(PRV), Bartha strain, into the uterus and Sanyinjiao(Sp6) of rats. After survival times of 4-5 days following the injection of PRV, the rats were perfused, and their brain and spinal cord were frozen sectioned($30\mu\textrm{m}$). These sections were stained by PRV immunohistochemical staining methods, and observed with light microscope. The results were as follows: 1. In the spinal cord, overlap areas of PRV labeled neurons projecting to uterus and Sp6 were observed in lamina VII, IX and X areas of cervical segments. In thoracic segments, overlap areas were observed in lamina IV, VII, X and intermediolateral n.. In lumbar segments, overlap area of PRV labeled neurons were observed in lamina I, V-VII, IX, X and intermediolateral n.. In sacral segments, overlap areas of PRY labeled neurons were observed in lamina N, V, VII, X and sacral parasympathetic n.. 2. In the brain, overlap areas of PR V labeled neurons projecting to the uterus and Sp6 were observed in lateral paragigantocellular n., rostroventrolateral reticular n., raphe obscurus n., raphe pallidus n., raphe magnus n., locus coeruleus n., Barrington's n., A5 cell group, central gray n., paraventricular hypothalamic n. and arcuate n. This results suggest that overlap areas of PRV labeled neurons of the spinal cord projecting to the uterus and Sp6 might be the first-order neurons related to the viscera-somatic sensory and sympathetic preganglionic neurons. PRV labeled neurons of the brain may be the second and third-order neurons response to the movement of smooth muscle of uterus. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitoring the internal environment. These overlap areas of spinal cord and brainmay be related to autonomic centers related to regulation of uterus.

• 대한수의학회지
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• 제39권4호
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• pp.797-805
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• 1999

T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

• 미생물학회지
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• 제52권2호
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• pp.140-147
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• 2016

동물유래성분을 원재료로 사용하는 의료기기는 원료물질 유래 바이러스가 오염될 가능성이 있기 때문에 생산과정 중 바이러스가 오염되지 않도록 하여야 한다. 또한 생산공정은 오염될 가능성이 있는 바이러스들을 불활화하거나 제거하는 공정을 포함하여야 한다. 본 연구를 통해 돼지유래 이종골을 원재료로 사용한 바이러스 안전성이 보증된 치과용 골이식재(THE Graft$^{(R)}$) 제조공정을 개발하였다. THE Graft$^{(R)}$ 제조공정은 30% 과산화수소수와 80% 에탄올을 각각 처리하여 지방을 제거하는 공정과 $1,300^{\circ}C$ 열처리 공정을 통해 콜라겐과 유기물을 제거하는 공정을 포함한다. 또한 최종적으로 생산된 hydroxyapatite 성분의 골이식재에 25 kGy의 감마선을 조사하여 감염성 위해인자를 불활화하는 공정을 포함한다. THE Graft$^{(R)}$의 형태학적 특성을 소유래 hydroxyapatite 성분의 골이식재인 Bio-Oss와 SEM과 TEM을 이용하여 비교한 결과 구조적 특성이 유사함을 확인하였다. $1,300^{\circ}C$ 열처리 공정과 25 kGy 감마선 조사 공정의 바이러스 불활화 효과를 평가하기 위해 transmissible gastroenteritis virus (TGEV), pseudorabies virus (PRV), porcine rotavirus (PRoV), porcine parvovirus (PPV)를 모델 바이러스로 선정하였다. $1,300^{\circ}C$ 열처리 공정에서 TGEV, PRV, PRoV, PPV 모두 검출한계 이하로 불활화되었으며, 바이러스 로그 감소 값은 각각 ${\geq}4.65$, ${\geq}5.81$, ${\geq}6.28$, ${\geq}5.21$이었다. 또한 감마선 조사 공정에서도 TGEV, PRV, PRoV, PPV 모두 검출한계 이하로 불활화되었으며, 바이러스 로그 감소 값은 각각 ${\geq}4.65$, ${\geq}5.87$, ${\geq}6.05$, ${\geq}4.89$이었다. 두 공정에서 TGEV, PRV, PRoV, PPV의 누적 바이러스 로그 감소 값은 각각 ${\geq}9.30$, ${\geq}11.68$, ${\geq}12.33$, ${\geq}10.10$이었다. 이상의 결과에 의하면, THE Graft$^{(R)}$ 제조공정은 바이러스 안전성 보증을 위한 충분한 바이러스 불활화 능력을 가지고 있는 것으로 판단된다.

• 동의생리병리학회지
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• 제23권4호
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• pp.805-817
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• 2009

This study was to investigate central localization of neurons projecting to the urinary bladder and urinary bladder-related acupoints(Wijung, $BL_{40}$) and neurons of immunoreactive to hormones and hormone receptors regulating urinary bladder function by using peudorabies virus(PRV). In this experiment, Bartha's strain of pseudorabies virus was used in rats to trace central localization of urinary bladder-related neurons and urinary bladder-related acupoints($BL_{40}$) which can regulate urinary system. PRV was injected into the urinary bladder and acupoints($BL_{40}$) related urinary system. After six days survival of rats, mainly common labeled neurons projecting to the urinary bladder and urinary bladder-related acupoints were identified in spinal cord, medulla, pons and diencephalon by PRV immunohistochemical staining method. First-order PRV labeled neurons projecting to urinary bladder and urinary bladder-related acupoints were found in the cervical, thoracic, lumbar and sacral spinal cord. Commonly labeled preganglionic neurons were labeled in the lumbosacral spinal cord and thoracic spinal cord. They were found in the lateral horn area(sacral parasympathetic nucleus and intermediolateral nucleus), lamina V-X, intermediomedial nucleus and dorsal column area. The area of sensory neurons projecting to urinary bladder and Wijung($BL_{40}$) was L5-S2 spinal ganglia and T12-L1 spinal ganglia, respectively. In the brainstem, the neurons were labeled most evidently and consistently in the nucleus of tractus solitarius, area postrema, dorsal motor nucleus of vagus nerve, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), C3 adrenalin cells, parapyramidal area(lateral paragigantocellular nucleus), locus coeruleus, subcoeruleus nucleus, A5 cell group, Barrington's nucleus and periaqueductal gray matter. In the diencephalon, PRV labeled neurons were marked mostly in the paraventricular nucleus and a few ones were in the lateral hypothalamic nucleus, posterior hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus, median eminence, perifornical nucleus, periventricular nucleus and suprachiasmatic nucleus. In cerebral cortex, PRV labeled neurons were marked mostly in the frontal cortex, 1,2 area, hind limb area, agranular insular cortex. Immunoreactive neurons to Corticotropin releasiing factor(CRF), Corticotropin releasiing factor-receptor(CRF-R), c-fos and serotonin were a part of labeled areas among the virus-labeled neurons of urinary bladder and Wijung($BL_{40}$). The commonly labeled areas were nucleus tractus solitarius, area postrema, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), locus coeruleus, A5 cell group, Barrington,s nucleus, arcuate nucleus, paraventricular nucleus, frontal cortex 1, 2 area, hind limb, and perirhinal(agranular insular) cortex. These results suggest that overlapped CNS locations are related with autonomic nuclei which regulate the functions of urinary bladder-relate organs and it was revealed by tracing PRV labeled neurons projecting urinary bladder and urinary bladder-related acupoints. These commonly labeled areas often overlap with the neurons connected with hormones and hormone receptors related to urination.

• Journal of Veterinary Science
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• 제22권6호
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• pp.87.1-87.12
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• 2021

Background: African swine fever virus (ASFV), classical swine fever virus (CSFV), and porcine reproductive and respiratory syndrome virus (PRRSV) are still prevalent in many regions of China. Co-infections make it difficult to distinguish their clinical symptoms and pathological changes. Therefore, a rapid and specific method is needed for the differential detection of these pathogens. Objectives: The aim of this study was to develop a multiplex real-time quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) for the simultaneous differential detection of ASFV, CSFV, and PRRSV. Methods: Three pairs of primers and TaqMan probes targeting the ASFV p72 gene, CSFV 5' untranslated region, and PRRSV ORF7 gene were designed. After optimizing the reaction conditions, including the annealing temperature, primer concentration, and probe concentration, multiplex qRT-PCR for simultaneous and differential detection of ASFV, CSFV, and PRRSV was developed. Subsequently, 1,143 clinical samples were detected to verify the practicality of the assay. Results: The multiplex qRT-PCR assay could specifically and simultaneously detect the ASFV, CSFV, and PRRSV with a detection limit of 1.78 × 10⁰ copies for the ASFV, CSFV, and PRRSV, but could not amplify the other major porcine viruses, such as pseudorabies virus, porcine circovirus type 1 (PCV1), PCV2, PCV3, foot-and-mouth disease virus, porcine parvovirus, atypical porcine pestivirus, and Senecavirus A. The assay had good repeatability with coefficients of variation of intra- and inter-assay of less than 1.2%. Finally, the assay was used to detect 1,143 clinical samples to evaluate its practicality in the field. The positive rates of ASFV, CSFV, and PRRSV were 25.63%, 9.36%, and 17.50%, respectively. The co-infection rates of ASFV+CSFV, ASFV+PRRSV, CSFV+PRRSV, and ASFV+CSFV+PRRSV were 2.45%, 2.36%, 1.57%, and 0.17%, respectively. Conclusions: The multiplex qRT-PCR developed in this study could provide a rapid, sensitive, specific diagnostic tool for the simultaneous and differential detection of ASFV, CSFV, and PRRSV.