In order to survey the occurrence of waterborne viruses in Korean surface water, a total of 192 water samples from July 2003 to January 2006 were collected and analyzed. The presence of waterborne viruses was investigated by total culturable virus assay (TCVA) using buffalo green monkey kidney (BGMK) cells. The results showed that 63 of 192 samples (32.8%) were positive for waterborne viruses with the average concentration of 3.1+/-18 most probable numbers (MPN)/100 L. The relationship between the occurrence of the viruses and the physicochemical environmental factors revealed that there was a significant correlation between the turbidity of water and the occurrence of the viruses. It was also noted that the water temperature might have some relationship with the occurrence of the viruses, as the frequency of the viruses was higher in low temperature or winter season. Therefore, the occurrence of waterborne viruses in Korean surface water might be affected by the physicochemical environmental factors such as turbidity and water temperature.
Buffaloes ; Cercopithecus aethiops ; Kidney ; Seasons ; Water*

The ocular vascular resistance of the ciliary choroidal network has been investigated on measurements of pulse amplitude(PA). The ocular pulse is derived from the ciliary choroidal blood flow which account for more than 90% of the total ocular blood flow(OBF). In 5 normal Cynomolgus monkeys with topical application(50 micro l) of 2% epinephrine, the PA revealed a significant reduction(0.32 +/- 0.06mmHg, mean +/- SEM, p<0.01). In the other 5 normal Cynormlgus monkeys with 4% pilocarpine, the PA revealed no significant increment(0.32 +/- 0.15mmHg, mean +/- SEM, p<0.10) by the OBF system. These results suggest that topical application of epinephrine produces a significant reduction of OBR but pilocarpine produces a mild increment of OBF.
Choroid ; Epinephrine* ; Haplorhini ; Macaca fascicularis ; Pilocarpine* ; Vascular Resistance

The ocular vascular resistance of the ciliary choroidal network has been investigated on measurements of pulse amplitude(PA). The ocular pulse is derived from the ciliary choroidal blood flow which account for more than 90% of the total ocular blood flow(OBF). In 5 normal Cynomolgus monkeys with topical application(50 micro l) of 2% epinephrine, the PA revealed a significant reduction(0.32 +/- 0.06mmHg, mean +/- SEM, p<0.01). In the other 5 normal Cynormlgus monkeys with 4% pilocarpine, the PA revealed no significant increment(0.32 +/- 0.15mmHg, mean +/- SEM, p<0.10) by the OBF system. These results suggest that topical application of epinephrine produces a significant reduction of OBR but pilocarpine produces a mild increment of OBF.
Choroid ; Epinephrine* ; Haplorhini ; Macaca fascicularis ; Pilocarpine* ; Vascular Resistance

AIMTo assess the contraceptive efficacy of the benzene chromatographic fraction of the chloroform extract of the seeds of Carica papaya in langur monkeys.

METHODSThe test substance was given p.o. to five monkeys at 50 mg/kg body weight/day for 360 days. Control animals (n=3) received olive oil as vehicle. Sperm parameters as per World Health Organization standards, sperm functional tests, morphology of testis and epididymis, haematology, clinical biochemistry, serum testosterone and libido were evaluated. Following completion of 360 days treatment the animals were withdrawn from the treatment and the recovery pattern was assessed by semen analysis and sperm functional tests.

RESULTSTotal inhibition of sperm motility was observed following 60 days of treatment that continued until 360 days study period. Sperm count, percent viability and percent normal spermatozoa showed a drastic decline following 30 days of treatment. Sperm morphology showed predominant mid piece abnormalities. Sperm functional tests scored in sterile range. Histology and ultrastructure of testis revealed vacuolization in the Sertoli cells and germ cells. Loss of cytoplasmic organelles was evident in spermatocytes and round spermatids. Histology and ultrastructure of epididymis of treated animals were comparable to those of control animals. Hematological and serum clinical parameters and testosterone levels fluctuated within the control range throughout the study period. Recovery was evident following 60-120 days of treatment withdrawal.

CONCLUSIONThe results suggest that the benzene chromatographic fraction of the chloroform extract of the seeds of Carica papaya shows contraceptive efficacy without adverse toxicity, mediated through inhibition of sperm motility.

Animals ; Benzene ; Carica ; Cercopithecidae ; Chloroform ; Chromatography ; Contraceptive Agents, Male ; pharmacology ; therapeutic use ; Male ; Models, Animal ; Phytotherapy ; Plant Extracts ; Seeds ; Sperm Motility ; drug effects ; Testis ; drug effects ; Testosterone ; blood

OBJECTIVETo study the preventive and therapeutic effects of recombinant IFN-alpha2b for nasal spray on SARS-CoV infection in Macaca mulata (rhesus monkey).

METHODSTen rhesus monkeys were divided into two groups, 5 in interferon group, and 5 in control group. Before and after SARS-CoV attack, the virus was detected in samples such as pharyngeal swab in all the two groups by Real-time PCR (RT-PCR) and virus isolation was performed.

RESULTSAfter virus attack, the level of SARS-CoV-specific IgG and neutralizing antibody were induced by SARS-CoV in the interferon group was weaker than in control group. Hematology items showed no apparent changes after virus attack in treated group. Through pathological examination, the morphology of the lung tissues of two Macaques in the treated group was normal, while the other three displayed the interstitial pneumonia with the thickened septum and infiltration with mononuclear cells. Among which, one monkey showed part of thickened septum fused with each other. These lesions in the interferon treated animals were similar to those seen in the animals in control group, but with smaller scope of pathological changes. No significant abnormity was detected in other organs.

CONCLUSIONRecombinant IFN-alpha2b could effectively interdict or weaken SARS-CoV injury in monkeys.

Animals ; Antiviral Agents ; therapeutic use ; Cercopithecus aethiops ; Disease Models, Animal ; Female ; Interferon-alpha ; therapeutic use ; Lung ; drug effects ; pathology ; virology ; Macaca mulatta ; Male ; Monkey Diseases ; drug therapy ; prevention & control ; virology ; Random Allocation ; Recombinant Proteins ; SARS Virus ; drug effects ; isolation & purification ; Severe Acute Respiratory Syndrome ; drug therapy ; prevention & control ; virology ; Vero Cells

To study the immunogenicity of recombinant adeno-asscociated virus type 1 expressing HIV-1 gp120 gene (rAAV2/1-gp120) in BALB/c mice and Rhesus macaques. The gp120 gene derived from Chinese HIV-1 isolates was constructed into rAAV2/1 and rAd5 vectors. Firstly, the immunogenicity of rAAV2/1-gp120 was compared with rAd5-gp120 in BALB/c mice when used once or twice in 3 weeks interval. Then the monkeys were immunized with rAAV2/1-gp120 once. The HIV-1 specific IgG levels and neutralization activity to pseudotyped HIV-1 virus were tested using ELISA and neutralization assay, and the cellular immune responses were analyzed by IFN-gamma enzyme-linked immunospot (ELISPOT) and in vivo CTL assays. Compared with rAd5-gp120 immunized mice, mice immunized with rAAV2/1-gp120 once in duced stronger gp120-specific IgG and were sustained for at least 21 weeks. rAd5-gp120 immunized mice generated stronger cellular immune responses than rAAV2/1-gp120 in spleen and draining lymph node. But only moderate gp120-specific in vivo CTL activity was observed in both rAAV2/1-gp120 and rAd5-gp120 immunized mice. Four of five monkeys vaccinated with rAAV2/1-gp120 generated gp120 specific IgG, the titer ranged from 1:100 to 1:400 with end-point dilution. Gp120 specific IgG could be detected 4 weeks after immunization and reached the peak at 10 weeks after immunization. No neutralization activity against pseudotyped HIV-1 virus expressing NL4-3 Env antigen was detected. In Conclusion, rAAV2/1-gp120 induced high level of HIV-1 specific IgG antibody and moderate cellular immune responses. No neutralizing antibody was elicited. It indicates that the env gene and immunization strategy should be optimized to elicit neutralizing antibody against HIV-1 in further studies.
AIDS Vaccines ; administration & dosage ; immunology ; Adenoviridae ; genetics ; Animals ; COS Cells ; Cercopithecus aethiops ; Cytotoxicity, Immunologic ; immunology ; Dependovirus ; genetics ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; HIV Antibodies ; immunology ; HIV Envelope Protein gp120 ; genetics ; immunology ; HIV-1 ; genetics ; immunology ; Immunization ; methods ; Immunoglobulin G ; immunology ; Macaca mulatta ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; administration & dosage ; genetics ; immunology ; Time Factors

The purpose of this research was to construct a lentiviral vector containing pir-b gene, and to detect the expression of pir-b gene in 293T cells. The open reading frame (ORF) of pir-b gene from the mRNA of mouse was cloned, then inserted into a sequencing vector. The pir-b gene was subcloned into the transfer plasmid of the lentivirus system, which was transfected together with the packaging plasmids into 293T cells by Lipofectin 2000, thereafter, the supernatant was collected and concentrated to transfect 293T cells. Western blot was used to detect the expression of the exogenous PIR-B after 293T cells were infected by the virus, while the lentivirus harboring egfp gene was packaged as the control group. The result indicated that the ORF of the pir-b gene was successfully cloned, the sequence of which was consistent to that was expected and the PIR-B protein could be expressed in 293T cells normally. It is concluded that the lentiviral vector containing pir-b gene was constructed successfully, which would contribute to illustrating the important role of pir-b gene in the immunological regulation.
Animals ; COS Cells ; Cercopithecus aethiops ; Genetic Vectors ; Lentivirus ; genetics ; Mice ; RNA, Messenger ; genetics ; Receptors, Immunologic ; genetics

OBJECTIVETo explore the driving force of the oral astringency produced by the interaction of theaflavin (TF) and epigallocatechin-3-gallate (EGCG) to human salivary a-amylase(HSA).

METHODSThe constants of the rate, equilibrium of reaction (k(a), k(d), K(A), and K(D)) and Langmuir, Freundlich adsorption isotherm (K(L), K(f), and Mm) were determined by surface plasmon resonance (SPR) technique and adsorption kinetics.

RESULTSBoth of Langmuir and Freundlich models could be used for describing the binding processes of TF and EGCG onto HSA surfaces, and there were no significant differences of the correlation coefficient of determination between these two models (P > 0.05). The constants of adsorption isotherm, the rate and equilibrium constants of the association for TF were higher than those of EGCG (P < 0.05). The rate and equilibrium constants of the dissociation for TF were lower than those of EGCG (P < 0.05). The affinity of TF to HSA was higher than that of EGCG.

CONCLUSIONThe sorely oral astringency is much easily produced by TF from the black tea rather than EGCG from the green tea. The driving force of the oral astringency is attributed to the hydrogen bonds of hydroxyl groups and hydrophobic interaction of galloyl groups in polyphenolic structures.

Adsorption ; Animals ; Antioxidants ; Biflavonoids ; Catechin ; analogs & derivatives ; Cercopithecus aethiops ; Humans ; Salivary alpha-Amylases ; Tea ; alpha-Amylases

OBJECTIVETo investigate the mitochondrial injury in enterovirus 71 (EV71)-infected Vero cells and explore the possible mechanism.

METHODSA clinical isolate of EV71 was inoculated to Vero cells and the EV71 antigen was detected by immunofluorescence assay. The morphological changes of Vero cells were observed using optical microscopy and transmission electron microscopy. The diameter and area density of the viral particles and the ratio and area density of vacuolated mitochondria in the cells were measured on the ultrastructural images.

RESULTSEV71-infected Vero cells underwent obvious changes and to a spherical morphology followed by cell death EV71 particles were detected in the cytoplasm by immunofluorescence. Ultrastructurally, the infected cells contained a large number of viral particles in the cytoplasm, with a clustered distribution and lattice-like arrangement. The diameter of the particles were 16.3 nm and the mean area density was 38.3%. Most of the mitochondria presented with swelling, vacuoles and degeneration. The ratio of the vacuolated mitochondria was 90.9% with a mean area density of 89.2%. Viral particles were also found in some mitochondria.

CONCLUSIONEV71 proliferates in the cytoplasm and invades the mitochondria of infected Vero cells leading to mitochondrial injury and cell death, suggesting that mitochondria are the targets for EV71 infection.

Animals ; Cercopithecus aethiops ; Cytoplasm ; virology ; Enterovirus ; Enterovirus Infections ; pathology ; Humans ; Mitochondria ; pathology ; virology ; Vero Cells ; virology

OBJECTIVEElectron microscopical study of infected cells to identify the pathogenic agent of SARS.

METHODSVero E6 cells infected with lung autopsy samples or nasopharyngeal swabs from SARS patients of Beijing and Guangzhou were inoculated. The supernatant and cultured cells exhibiting identifiable cytopathic effect (CPE) were prepared for electron microscopic study.

RESULTSExamination of CPE cells on thin-section revealed characteristic coronavirus particles within the cisternae of endoplasmic reticulum, Golgi apparatus, vesicles and extracellular space. They were mainly spherical or oval in shape, annular or dense, about 80 nm in diameter. Negative-stain electron microscopy identified coronavirus particles in culture supernatant, 80 - 120 nm in diameter, with club-shaped surface projections. Elongated, rod-, kidney- or other irregular shaped virons with the size of 100 - 200 nm by 60 - 90 nm were also found in the cultured cells infected with the lung samples from the Guangdong patients. Infectious virons entered cells by endocytosis or membrane fusion and released through a budding process.

CONCLUSIONThese data indicate a novel coronavirus as the causative agent of SARS. Most viral particles showed typical characteristics of coronavirus. The potential role of special shape viruses is expected to be further investigated.

Animals ; Cercopithecus aethiops ; Humans ; Microscopy, Electron ; SARS Virus ; ultrastructure ; Severe Acute Respiratory Syndrome ; virology ; Vero Cells