Anti-influenza properties of herbal extract of Althaea rosea in mice / 대한수의학회지

Althaea rosea has been used in traditional Chinese medicine to treat numerous diseases, but no studies have investigated its anti-influenza properties to date. In this study, we investigated the anti-influenza effects of Althaea rosea. BALB/c mice orally pretreated with Althaea rosea (200 µL, 0.1 mg/mL concentration in phosphate-buffered saline) and followed by infection of influenza A virus nasally showed higher survivability and lower lung virus titer against divergent subtypes of influenza A virus infection. We also found that oral administration of Althaea rosea elicited antiviral innate immune responses in serum, bronchoalveolar lavage fluid, small intestinal fluid, and the lungs. Taken together, these findings suggest that aqueous extracts of Althaea rosea are a potential candidate for use as an anti-influenza drug.

Anti-influenza properties of herbal extract of Althaea rosea in mice

Althaea rosea has been used in traditional Chinese medicine to treat numerous diseases, but no studies have investigated its anti-influenza properties to date. In this study, we investigated the anti-influenza effects of Althaea rosea. BALB/c mice orally pretreated with Althaea rosea (200 µL, 0.1 mg/mL concentration in phosphate-buffered saline) and followed by infection of influenza A virus nasally showed higher survivability and lower lung virus titer against divergent subtypes of influenza A virus infection. We also found that oral administration of Althaea rosea elicited antiviral innate immune responses in serum, bronchoalveolar lavage fluid, small intestinal fluid, and the lungs. Taken together, these findings suggest that aqueous extracts of Althaea rosea are a potential candidate for use as an anti-influenza drug.

Immunogenicity of a Canine Parvovirus 2 Capsid Antigen (VP2-S1) surface-expressed on Lactobacillus casei / 동물의과학연구지

Canine parvovirus (CPV2) is one of the most virulent virus causing acute hemorrhagic enteritis and myocarditis in dogs. Infection mainly caused by the ingestion of virus through the mucosal route. Therefore, induction of mucosal immunity is essential in prevention of Canine Parvovirus (CPV2) infection. For safe and effective delivery of viral antigens to the mucosal immune system, a novel surface antigen display system for lactic acid bacteria using the poly-gamma-glutamic acid synthetase A protein (pgsA) of Bacillus subtilis as an anchoring matrix was applied in order to display CPV2 antigen on the surface of the recombinant L. casei. Recombinant fusion proteins comprised of pgsA and the capsid protein (VP2-S1) showed stable expression in Lactobacillus casei. Surface localization of the fusion protein was verified by cellular fractionation analyses. Oral and nasal inoculations of recombinant L. casei into mice resulted in high levels of serum immunoglobulin G (IgG) and mucosal IgA, as demonstrated by ELISA using recombinant VP2-S1 proteins. Mice receiving intranasal immunization mounted higher antibody response than those receiving oral immunization. These results indicate that mucosal immunization with recombinant L. casei expressing CPV2 VP2-S1 protein on its surface provides an effective means for elicitation of strong antibody responses against CPV 2 VP2-S1.

Transanal Endoscopic Microsurgery for the Treatment of Well-Differentiated Rectal Neuroendocrine Tumors

PURPOSE: Recently, an increase in well-differentiated rectal neuroendocrine tumors (WRNETs) has been noted. We aimed to evaluate transanal endoscopic microsurgery (TEM) for the treatment of WRNETs. METHODS: Between December 1995 and August 2009, 109 patients with WRNETs underwent TEM. TEM was performed for patients with tumors sizes of up to 20 mm and without a lymphadenopathy. These patients had been referred from other clinics after having been diagnosed with WRNETs by using a colonoscopic biopsy; they had undergone a failed endoscopic submucosal dissection (ESD) or endoscopic mucosal resection (EMR) and exhibited an involved resection margin and remaining tumor after ESD or EMR, regardless of the distance from the anal verge. This study included 38 patients that had more than three years of follow-up. RESULTS: The mean age of the patients was 51.3 +/- 11.9 years, the mean tumor size was 8.0 +/- 3.9 mm, and no morbidity occurred. Thirty-five patients were asymptomatic. TEM was performed after a colonoscopic resection in 13 cases because of a positive resection margin, a residual tumor or a non-lifting lesion. Complete resections were performed in 37 patients; one patient with a positive margin was considered surgically complete. In one patient, liver metastasis and a recurrent mesorectal node occurred after five and 10 years, respectively. CONCLUSION: TEM might provide an accessible and effective treatment either as an initial or as an adjunct after a colonoscopic resection for a WRNET.

Actinomycosis of the Appendix: A Case Report

Actinomycosis is an uncommon disease caused by actinomycoses, which is a normal flora in the human mucosal membrane. It is difficult to diagnose pre- and intra-operatively and requires long-term use of antibiotics even after surgery. Especially, abdominal actinomycosis is frequently misdiagnosed as a tumor, diverticulitis, chronic inflammatory disease, or other infectious disease preoperatively. Thus, we report the case of a 21-yr-old male patient who was thought to have acute appendicitis and who underwent a cecal wedge resection, including the appendix, with the assistance of laparoscopy for appendiceal actinomycosis.

Activation of the intrinsic mitochondrial apoptotic pathway in swine influenza virus-mediated cell death

The mitochondrial pathway of swine influenza virus (SIV)-induced apoptosis was investigated using porcine kidney (PK-15) cells, swine testicle (ST) cells, and HeLa cervical carcinoma cells which are known not to support viral replication. As judged by cell morphology, annexin V staining, and DNA fragmentation, PK-15 and ST cells infected with three different subtypes of SIV (H1N1, H3N2, and H1N2) were obviously killed by apoptosis, not necrosis. SIV infection in PK-15 and HeLa cells was shown to decrease the cellular levels of Bcl-2 protein compared to that of mock-infected control cells at 24 h post-infection, whereas expression levels of Bax protein increased in the PK-15 cells, but did not increase in HeLa cells by SIV infection. Cytochrome c upregulation was also observed in cytosolic fractions of the PK-15 and HeLa cells infected with SIV. Apoptosome (a multi-protein complex consisting of cytochrome c, Apaf-1, caspase-9, and ATP) formation was confirmed by immunoprecipitation using cytochrome c antibody. Furthermore, SIV infection increased the cellular levels of TAJ, an activator of the JNK-stressing pathway, and the c-Jun protein in the PK-15 and HeLa cells. Taken together, these results suggest that the mitochondrial pathway should be implicated in the apoptosis of PK-15 cells induced by SIV infection.

Isolation and Genetic Characterization of H1N2 Subtype of Influenza A Virus from Pigs in Korea

In order to investigate swine influenza virus (SIV) infection in South Korea, 180 nasal swabs from pigs with respiratory symptoms of 18 different farms were collected between November 2001 and February 2002. Of the 180 swabs, 52 were positive for SIV by a multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay. Positive samples were then subtyped by two multiplex RT-PCR assays, and 43 and 9 samples were found to be H1N2 and H3N2, respectively. SIV was isolated from the positive samples using embryonated chicken eggs. Two H1N2 isolates designated as A/Sw/ Korea/CY01/02 and A/Sw/Korea/CY02/02 were genetically characterized. Comparison of the nucleotide sequences between the two isolates showed 99.6% to 99.9% nucleotide identity of each gene segment. Pairwise sequence analysis of the hemagglutinin (HA)1 gene segments indicated that HA genes of the Korean isolates were more closely related to those of the US H1N2 isolates (94.5% to 97.9% amino acid identity) than those of the Japanese H1N2 isolates (82.4% to 93.7% amino acid identity). Amino acid sequences of the Korean isolate (A/Sw/Korea/CY02/02) showed high homology with the US H1N2 isolates; neuraminidase (NA) (97.4~99.2%), matrix (98.8~99.9%), nucleoprotein (97.6~ 98.0%), non-structural (96.8~98.6%), PA (97.9~98.7%), PB1 (98.3~99.6%) and PB2 (97.8~98.9%) genes.

Sequence analysis of canine LINE-1 elements and p53 gene in canine transmissible venereal tumor

LINEs (long interspersed nuclear elements or long interspersed repeated DNA elements) contains two open reading frames (ORFs), ORF1 and ORF2. We analysed the ORF2 located in the 5' region to the first exon of oncogene c-myc in canine transmissible venereal tumor (TVT) cell. We also showed the transcription activation was induced by this TVT-LINE sequence using CAT assay. To identify the mutation of tumor suppressor gene, sequence analysis of p53 from TVT cell was performed. We identified the point mutation of 964 nucleotide (T-->C) resulting in the change of amino acid (Phe-->Ser) of p53 tumor suppressor protein.

Seroprevalence of Antibody to Procine Reproductive and Respiratory Syndrome Virus in Diagnostic Submissions

During the period from January to December of 2001, a total of 3,391 swine sera were submitted to our laboratory from 256 farms for the diagnosis of porcine reproductive and respiratory syndrome (PRRS). The antibody to porcine reproductive and respiratory syndrome virus (PRRSV) was tested by the indirect immunofluorescent antibody (IFA) test. Of the 256 farms tested, 230 farms (89.8%) were positive for the PRRSV antibody. The overall seroprevalence of the PRRSV antibody was 52.1% (1765/3391). Most of the pigs seemed to be infected with PRRSV at around 50 to 60 days old. The seroprevalence of the antibody became higher with age, and peaked at around 100 days old. More than one-third of the adult pigs, including boars, gilts, and sows, was positive for the PRRSV antibody. The infection of PRRSV was chronic and confined to growers and/or finishers in most farms. However, the antibody was detected in all production phases at some farms.

Media Role in Health Policy

No abstract available.

Expression of Open Reading Frame 5 Protein of Porcine Reproductive and Respiratory Syndrome Virus Using Semliki Forest Virus Expression System

The ORF5 gene encodes a major envelope glycoprotein (GP5), which is one of the three major proteins of porcine reproductive and respiratory syndrome virus (PRRSV). The GP5 protein has been known to be a 24.5-26kDa N-glycosylated envelope protein. The GP5 is involved in inducing neutralizing antibodies. For this reason, the GP5 is primary candidate for the PRRSV subunit vaccine. To produce the native form of GP5 in mammalian cells, we have cloned the ORF5 gene from PRRSV CNV-1 into the Semliki Forest virus (SFV)-based expression vector, resulting in recombinant pSFV-ORF5. By the infection with recombinant pSFV-ORF5 to BHK-21 cells, the GP5 expression was confirmed by immunocytochemistry and immunoblotting assay. The recombinant virus particle harboring ORF5 gene was infectious to BHK-21 and MARC-145. The RNA synthesis and expression of GP5 in the infected cell was also confirmed by RT-PCR.

Expression of Angiostatin Using DNA-Based Semliki Forest Virus Replicon

Angiogenesis is recognized as a critical factor in the growth of tumor cells and plays a key role in the tumor metastasis. Recent studies for antiangiogenic substances are getting popular. The angiostatin, one of the antiangiogenic substances, leads to the increased apoptosis of the tumor cells by inhibiting the neovascularization of the tumor. The angiostatin was identified as the internal fragments of the plasminogen which has no antiangiogenic activity. By hydrolysis of the plasminogen, the angiostatin can be produced. In this study, we constructed the SFV-derived DNA vector by employing the cytomegalovirus immediate early enhancer/ promoter (CMV). This vector makes it possible to transfect the cells with DNA without the in vitro transcription process. The C-myc epitope and polyhistidine residue sequences were placed in downstream of the angiostatin gene to make it eligible to detect the expressed protein. The murine Ig kappa-chain V-J2-C signal sequence was placed in upstream to secrete the expressed protein from the cells. We confirmed the expression of angiostatin in the BHK-21 cells using DNA-based SFV replicon.

'Mad Cow Disease' in the Korean Newspapers

No abstract available.

Immunologic reactivity of a lipopolysaccharide-protein complex of type A Pasteurella multocida in mice

The immunologic reactivity of a lipopolysaccharide (LPS)-protein complex isolated from a potassium thiocyanate extract of a Pasteurella multocida (capsular type A and somatic type 3) strain was evaluated in mice. The LPS-protein complex provided 100% protection in mice against a challenge with the homologous strain. However, when the complex was fractionated into LPS and protein moieties by phenol-water treatment, both components lacked immunogenicity. The complex and extracted components were mitogenic for mouse B lymphocytes with the protein moiety the most active. Although immune serum against the LPS-protein complex protected mice against challenge thereby indicating a role for humoral immunity, the LPS-protein complex of P. multocida was also found to induce cell-mediated immunity. This cell-mediated immunity was demonstrated in mice immunized with the complex by: (1). mitogenic responses of T lymphocytes, (2). induction of delayed type hypersensitivity reaction in the hind footpads, and (3). enhanced resistance to challenge infection with Salmonella enteritidis.

Transurethral Exchange of Double-J Ureteral Stent Using Goose-Neck Snare

PURPOSE: To evaluate the usefulness of transurethral exchange of double-J ureteral stent as an effective alternative to the cystoscopic approach. MATERIALS AND METHODS: There were 20 exchange cases involving seven patients (six women and one man) who initially underwent antegrade manipulation of a double-J ureteral stent. Indications for stent placement were ureteral stricture caused by malignancy in six patients [cervical carcinoma (n=5), stomach carcinoma (n=1) ], and renal tuberculosis in one. An 8-F Nelaton catheter was inserted in the bladder via the urethra and contrast material was injected until the bladder was fully distended. The distal end of a double-J ureteral stent was extracted to the urethral orifice using a goose-neck snare and a 0.035 "stiff guide wire was then advanced to the renal pelvis through the stent. After that, the stent was removed and a 4-F Cobra catheter was advanced to the renal pelvis along the guide wire. Contrast material was injected through the catheter, and the renal pelvis, calyx and ureter were opacified. The 0.035 "stiff guide wire was again inserted via the catheter, a new double-J ureteral stent was inserted, and the catheter removed. Finally, the new double-J stent was properly located within the renal pelvis and the bladder. RESULTS: Double-J ureteral stents were successfully exchanged in 19 of 20 exchange cases. After the procedure, all patients reported tolerable, minimal lower abdominal pain. CONCLUSION: Transurethral exchange of double-J ureteral stent is a useful alternative to cystoscopy.

CT Square Index of Hepatic Segment: The Significance in Cirrhotic Patients

PURPOSE: To determine changes in the square index of the liver segments of liver cirrhosis(LC) patients, as seen on CT, and the value of this indicator during follow-up. MATERIALS AND METHODS: Twenty-three patients with LC were included in this study. Abdominal CT scans were performed twice in each patient and the mean follow-up period was 15 (6-36) months. We measured the square index of the right lobe, the caudate lobe, and the medial and lateral segment of the left lobe of the liver, as seen on initial and follow-up CT images, and compared the results. The square index was obtained by deter-mining the product of the transverse and longitudinal diameters. According to the Child-Pugh classification, the condition was classified as either progressive or non-progressive, and the square index was compared between the two groups. RESULTS: The square index of the left lobe medial segment showed a significant decrease in both the progression group(n=13) and non-progression group(n=10), while that of the right lobe was significantly lower only in the progression group. There was no significant change in the square index of the caudate lobe or the lateral segment of the left lobe. CONCLUSION: For predicting the progression of LC, the square index of the medial segment of the left lobe is a more sensitive index than the Child-Pugh classification. For ascertaining the progression of the condition, the square index of the right lobe is a valuable deferminant.

Zebra Pattern in CT during Arterial Portography: Analysis of Associated Factors

PURPOSE: To analyze the factors associated with the zebra pattern in CT during arterial portography(CTAP). MATERIALS AND METHODS: In 275 CTAP procedures, the factors associated with the zebra pattern, such as laminar flow in the portal vein, the presence of liver cirrhosis, the artery selected for CTAP, location of the catheter tip in the superior mesenteric artery(SMA), splenic volume, and the existence of an aberrant right hepatic artery(RHA) emerging from the SMA were analyzed. RESULTS: In 106 of 275 procedures (38.5%), a zebra pattern was apparent. Portal venous laminal flow was seen in 92 % of procedures in the group with this pattern and in 63 % in the group without it. Eighty-three of 235 procedures (35.3 %) in which the SMA was injected and 23 of 40(57.5 %) involving splenic artery injection showed the zebra pattern. In 22 of 35(62.8 %) in which the catheter tip was located in the distal SMA and 61 of 200 ( 30.5 %) in which this was at a proximal site, the zebra pattern was evident. Mean splenic volume was less in the group with the zebra pattern. The effect on the zebra pattern of liver cirrhosis and an aberrant RHA emerging from the SMA was not statistically significant. CONCLUSION: In CTAP, the incidence of the zebra pattern was 38.6%, and was related to laminal flow in the portal vein. The pattern is frequently seen in CTAP involving contrast injection via the splenic artery, distal location of a catheter tip in the SMA, and small splenic volume.

Mucinous and Nonmucinous Type of Bronchioloal veolar Carcinoma: Difference in CT Findings

PURPOSE: To search for CT findings which helpfully differentiate mucinous from nonmucinousbronchi-oloalveolar carcinoma and to assess the difference in stages between the two types of tumors. Twenty-two patients with pathologically proven bronchioloalveolar carcinoma (BAC) were included inthis study. On the basis of CT findings, tumors were classified as either solitary or multiple and as eithermass/nodule, consolidation, or mixed type. CT stages of the tumors were determined by two radiologists andconclusions were reached by consensus. RESULTS: Twelve patients had nonmucinous BACs and ten had mucinous BACs.Among the ten cases of mucinous BAC, six were solitary and four were multiple. These were mass/nodule (n=3),consolidation (n=5), and mixed pattern (n=2). In contrast, among the twelve cases of nonmucinous BAC, six weresolitary and six were multiple. All were mass/nodule, except for one mixed type. Among the mucinous BACs, threewere operable and seven (above stage IIIa) were inoperable. Among the nonmucinous BACs, four were operable andeight were inoperable. CONCLUSION: Consolidation was more common in mucinous BAC and mass/nodule was more commonin non-mucinous BAC (p0.05).

Prevalence and Identification of Rotaviruses in Stool Specimens of Patients with Acute Diarrhea from Several Regions of Korea

PURPOSES: Rotaviruses are the main cause of infantile diarrheal diseases worldwide. The purpose of this study is to obtain epidemiologic data of rotavirus infections in Korea. METHODS: Stool specimens were collected from 150 patients with acute diarrheal symptoms, who were admitted to Yonsei Medical Center and Chungbuk National University Hospital. After isolating the virus from the specimens, the viruses were identified as rotaviruses by electron microscope and fluorescent microscope after staining with rotavirus VP6-specific monoclonal antibody. RNA was extracted from the specimens by modified phenol/chloroform method. Electropherotying was done with extracted RNA samples after silver staining of the gels. Rotavirus serotyping was done using commercial serotyping ELISA kit. RESULTS: Rate of infection from rotavirus stool samples was 60%. Isolated rotaviruses were all serogroup A and a majority(46.0%) of these viruses were long type. Serotypes 1 and 2 were identified with serotype 1 being the majority(52.2%). There were no differences in these types between 1991 and 1992, and also between Seoul and Cheongju area. CONCLUSION: Electropherotype of rotavirus epidemic in Korea was serogroup A, and long type in the majority. Serotypes of rotavirus epidemic in Korea were type 1 and type 2. These results show that the rotavirus reassortant vaccine developed in America could also handle the rotaviral disease in Korea.

Longevity of Antibodies to Live Orientia tsutsugamushi Inoculated in Sprague Dawley Rats

In Sprague Dawley (SD) rats, antibodies against strains of Orinentia tsutsugamushi, Kato, Karp and Gilliam, were produced in order to investigate their longevity and cross-reactivities to their corresponding homologous and heterologous antigens. By immunofluorescence assay (IFA) of IgG and IgM, it was shown that the immunity to the homologous strains persisted at a higher level (longevity of at least 34 weeks with higher IFA titers). On the other hand, the immunity to the heterologous strains persisted at a lower level (longevity of 10 to 34 weeks with lower IFA titers). Since infection with one strain of O. tsutsugamushi does not preclude reinfection with other strains, understanding of the antigenic diversity of O. tsutsugamushi and duration of the immunity to both homologous and heterologous strain is very important in diagnosis of scrub typhus.