This study was intended to establish a method for preparation of purified and viral-inactivated placenta hemoglobin. The optimum preparative condition resulted in the up-grading of purity,recovery and so on. A quality control was also established for the purification of hemoglobin. Compared to present purification methods, this method is easy to operate, needs low investment and running cost, and has the advantage of simultaneous operation for purification and viral-inactivation. The resulted hemoglobin had high purity and recovery, and the physicochemical property measured up to that in international reports. So this method is suitable for preparing purified and viral-inactivated hemoglobin on an adequate scale, and is useful for further development of blood substitutes.
Blood Substitutes ; Fetal Blood ; chemistry ; virology ; Hemoglobins ; isolation & purification ; Humans ; Virus Inactivation

This study was aimed to evaluate the efficacy of riboflavin photochemical inactivation of virus in red blood cells by using animal models. human cytomegalovirus (HCMV) plus red blood cells were used as indicator, 30 BALA/c mice were divided into the experimental group (n = 10), virus control group (n = 10), visible light control group (n = 5) and red blood cell control group (n = 5). Mice in experimental group were inoculated with red blood cells inactive by the riboflavin photochemical, mice in virus control group was injected with red blood cells without riboflavin photochemical inactivation treatment, and mice in light control group was infused with red blood cells irradiated by visible light, and mice in red blood cells control group was injected with normal red blood cells. The virus was isolated in vitro from mice of various groups, the HCMV UL83 gene was detected by PCR, the PP65 antigen was identified by indirect immunofluorescence. The results indicated that the virus isolation, PCR detection and indirect immunofluorescence identification all showed positive in virus control group and visible light control group, while the results of detection in experimental and red blood cell control groups were negative. It is concluded that riboflavin photochemical viral inactivation of red blood cells is effective.
Animals ; Erythrocytes ; virology ; Humans ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Photochemistry ; Riboflavin ; pharmacology ; Virus Inactivation

The effective inactivation of microorganisms in drinking water by Ultraviolet (UV) irradiation is regarded as a new low-cost water treatment method shoeing high removal rate of relatively stable infectious virus particles including poliovirus. In the present study, we examined virus inactivation by UV in various water environments. Samples were collected from finished water and surface water, and tested for turbidity. UV dose of 18, 22, 30, 36 and 40 milli-Joule (mJ)/cm2 were used by combination of 2 mW/cm2 UV intensity and time of 9, 11, 15, 18 and 20 second. Depths of water were fixed at 0.37 cm and 8 cm, and virus titers were shown by plaque forming unit (PFU). Poliovirus was inactivated to 99.0% by 18 mJ/cm2 of UV dose in the condition of 0.08 Nephelometry Turbidity Unit (NTU) and 8 cm depth of water. Poliovirus at 30 mJ/cm2 of UV dose under the same condition was inactivated to 99.7%. Furthermore, Poliovirus at 56.60 NTU and 8 cm depth of water was inactivated to 92.0% and 98.5% by 18 mJ/cm2 and 30 mJ/cm2 of UV dose, respectively. The degrees of virus inactivation were dependent upon the UV dose, the turbidit, y and the depth of water. In conclusion, introduction of UV disinfections can be considered in drinking water purification systems in case reasonable engineering support is possible.
Drinking Water ; Nephelometry and Turbidimetry ; Poliovirus ; Shoes ; Virion ; Virus Inactivation ; Water Purification ; Water*

The effective inactivation of microorganisms in drinking water by Ultraviolet (UV) irradiation is regarded as a new low-cost water treatment method shoeing high removal rate of relatively stable infectious virus particles including poliovirus. In the present study, we examined virus inactivation by UV in various water environments. Samples were collected from finished water and surface water, and tested for turbidity. UV dose of 18, 22, 30, 36 and 40 milli-Joule (mJ)/cm2 were used by combination of 2 mW/cm2 UV intensity and time of 9, 11, 15, 18 and 20 second. Depths of water were fixed at 0.37 cm and 8 cm, and virus titers were shown by plaque forming unit (PFU). Poliovirus was inactivated to 99.0% by 18 mJ/cm2 of UV dose in the condition of 0.08 Nephelometry Turbidity Unit (NTU) and 8 cm depth of water. Poliovirus at 30 mJ/cm2 of UV dose under the same condition was inactivated to 99.7%. Furthermore, Poliovirus at 56.60 NTU and 8 cm depth of water was inactivated to 92.0% and 98.5% by 18 mJ/cm2 and 30 mJ/cm2 of UV dose, respectively. The degrees of virus inactivation were dependent upon the UV dose, the turbidit, y and the depth of water. In conclusion, introduction of UV disinfections can be considered in drinking water purification systems in case reasonable engineering support is possible.
Drinking Water ; Nephelometry and Turbidimetry ; Poliovirus ; Shoes ; Virion ; Virus Inactivation ; Water Purification ; Water*

BACKGROUND: Tego-51(R), one of the amphoteric surfactants, has been considered as an effective disinfectant having both bactericidal and fungicidal effect. The author evaluated inactivation effect of Tego-51(R) on viruses causing disease in humans. METHODS: Influenza virus B, respiratory syncytial virus (RSV), herpes simplex virus 1 (HSV-1), adenovirus and echovirus 30 were exposed to diluted Tego-51(R) solution (1% and 0.1%) for 5, 10 and 30 minutes respectively and were inoculated onto the following cells: Influenza virus B, MDCK; RSV, HEp-2; HSV-1, HEp-2; adenovirus, Vero; and echovirus 30, RD. After incubation for 5 to 6 days, viral infection was identified with indirect immunofluorescent methods. RESULTS: Influenza virus B, RSV and HSV-1 which are enveloped viruses were inactivated after exposure of the viruses to Tego-51(R) for 5 minutes. Non-enveloped adenovirus and echovirus 30 were not inactivated after exposure for 30 minutes. CONCLUSIONS: Tego-51(R) appears to be effective in inactivation of enveloped viruses at concentrations used for disinfection of pathogenic bacteria and fungi.
Adenoviridae ; Bacteria ; Disinfection ; Enterovirus B, Human ; Fungi ; Herpesvirus 1, Human ; Humans ; Orthomyxoviridae ; Respiratory Syncytial Viruses ; Surface-Active Agents ; Virus Inactivation*

To determine whether addition of vitamin C (Vit C) to single-unit plasma could influence the efficacy of inactivating viruses and could maintain the activity of plasma proteins by methylene blue (MB)-light treatment. Vesicular stomatitis virus (VSV) Indiana strain was used as the indicating virus. Human plasma containing VSV was added with different concentrations of Vit C and final concentration 1 micromol/L MB and irradiated by fluorescence at an intensity of 40,000 lx, samples were collected at different times for detection. Cytopathic effect was used to test the effect of virus inactivation. A segment of the nucleic acid encoding capsid protein of VSV was amplified with RT-PCR. Some methods, such as the Clauss method, the one-stage method, microimmunoelectrophoresis, were used to investigate the changes of plasma components. The results showed that when the VSV plasma was added with 240 micromol/L Vit C and treated by MB-light irradiation for 60 min, the titer of VSV decreased by more than 8 lg TICD50/ml. Meanwhile, target segment amplification of VSV was also negative. The recovery rates of fibrinogen and coagulation factor VIII (FVIII: C) were 83.55% and 81.67% respectively, which had significant difference comparing with the routine MB-fluorescent light treatment. Most of plasma proteins were not affected significantly. No change in immunogenicity of these proteins was observed by using microimmunoelectrophoresis. It is concluded that virus inactivation is not influenced and plasma proteins are effectively protected by Vit C. Vit C can be used as a protector and is beneficial to improving the quality of plasma subjected to MB- photodynamic treatment.
Ascorbic Acid ; pharmacology ; Blood Proteins ; metabolism ; Humans ; Light ; Methylene Blue ; pharmacology ; Plasma ; virology ; Vesicular stomatitis Indiana virus ; drug effects ; Virus Inactivation ; drug effects

OBJECTIVEThis paper aims to investigate the apoptotic effect of inactivated Sendai virus (hemagglutinating virus of Japan-enveloped, HVJ-E) on murine melanoma cells (B16F10) and the possible mechanisms involved in the putative apoptotic reactions.

METHODSB16F10 cells were treated with HVJ-E at various multiplicities of infection (MOI), and the reactive oxygen species (ROS), cell viability, and apoptosis were measured. Next, the roles of ROS in the regulation of Bcl-2/Bax and the activation of mitogen-activated protein kinase (MAPK) pathways in HVJ-E-treated B16F10 cells were analyzed. To further evaluate the cytotoxic effect of HVJ-E-generated ROS on B16F10 cells, HVJ-E was intratumorally injected, both with and without N-acetyl-L-cysteine (NAC), into melanoma tumors on BALB/c mice. Tumor volume was then monitored for 3 weeks, and the tumor proteins were separated for immunoblot assay.

RESULTSTreatment of B16F10 cells with HVJ-E resulted in a dose-dependent inhibition of cell-viability and an induction of apoptosis. The latter effect was associated with the generation of ROS. Inhibition of ROS generation by NAC resulted in a significant reduction of HVJ-E-induced Erk1/2, JNK, and p38 MAPK activation. Additionally, ROS inhibition caused a decrease in the Bcl-2/Bax ratio as well as promoting activation of apoptosis both in vitro and in vivo.

CONCLUSIONThese results suggest that HVJ-E possesses potential anticancer activity in B16F10 cells through ROS-mediated mitochondrial dysfunction involving the MAPK pathway.

Animals ; Apoptosis ; Cell Line, Tumor ; Mice ; Mitogen-Activated Protein Kinase 1 ; genetics ; metabolism ; Reactive Oxygen Species ; metabolism ; Respirovirus Infections ; virology ; Sendai virus ; physiology ; Virus Inactivation

OBJECTIVEThe current study aims to investigate the effect of Hemagglutinating virus of Japan envelope (HVJ-E) on induction of apoptosis and autophagy in human prostate cancer PC3 cells, and the underlying mechanisms.

METHODSPC3 cells were treated with HVJ-E at various multiplicity of infection (MOI), and the generated reactive oxygen species (ROS), cell viability, apoptosis, and autophagy were detected, respectively. Next, the role of ROS played in the regulation of HVJ-E-induced apoptosis and autuphagy in PC3 cells were analysed. In the end, the relationship between HVJ-E-induced apoptosis and autuophagy was investigated by using rapamycin and chloroquine.

RESULTSFlow cytometry assay revealed that HVJ-E treatment induced dose-dependent apoptosis and that the JNK and p38 MAPK signaling pathways were involved in HVJ-E-induced apoptosis in PC3 cells. In addition, HVJ-E was able to induce autophagy in PC3 cells via the class III PI3K/beclin-1 pathway. The data also implyed that HVJ-E-triggered autophagy and apoptosis were ROS dependent. When ROS was blocked with N-acetylcysteine (NAC), HVJ-E-induced LC3-II conversion and apoptosis were reversed. Interestingly, HVJ-E-induced apoptosis was significantly increased by an inducer of autophagy, rapamycin pretreatment, both in vitro and in vivo.

CONCLUSIONHVJ-E exerts anticancer effects via autophagic cell death in prostate cancer cells.

Apoptosis ; physiology ; Autophagy ; physiology ; Cell Line, Tumor ; Cell Survival ; Humans ; Male ; Oncolytic Virotherapy ; Prostatic Neoplasms ; metabolism ; Reactive Oxygen Species ; metabolism ; Sendai virus ; immunology ; physiology ; Virus Inactivation

OBJECTIVETo study the method for cultivation and inactivation of SARS-CoV.

METHODSIn order to choose the sensitive cell strain and the best infection dose of the virus, Vero, Vero-E6 and 2BS cell lines were infected with SARS-CoV. The cultivation temperature was selected among 25 degrees C, 33 degrees C and 37 degrees C. The best inactivating time and effect were observed with beta-propiolactone whose concentration ranged from 1:2000 to 1:20,000 at room temperature.

RESULTSVero and Vero-E6 cell lines were sensitive to SARS-CoV. The cytopathic changes of the cells were 75% at 37 degrees C in 5 percent CO2 incubator after infection. SARS-CoV was inactivated completely in beta-propiolactone (1:4000). The toxicity of beta-propiolactone was hydrolyzed completely when the inactivated virus was cultured for 16 hours at 2 degrees C, 8 degrees C and in water bath for 2 hours at 37 degrees C.

CONCLUSIONThe titer of SARS-CoV was the highest when it was cultured in Vero or Vero-E6 cells for 72 hours at 37 degrees C in 5 percent CO2 incubator. SARS-CoV was inactivated completely in beta-propiolactone when its concentration was 1:4000 and the interaction time was 1 hour.

Animals ; Cercopithecus aethiops ; Dose-Response Relationship, Drug ; Propiolactone ; pharmacology ; SARS Virus ; drug effects ; growth & development ; Temperature ; Time Factors ; Vero Cells ; Virus Inactivation ; drug effects

Four out of 10 patients of X-linked severe combined immunodeficiency (X-SCID) were finally developed leukemia after receiving the treatment of gene therapy delivered by gamma-retroviral vectors. This is due to the vector integrated to the proximity of lmo2 etc proto-oncogene promoters, leading to the activation of onco-gene expression, which raises the concern of the bio-safety of gene therapy vectors. Lentiviral vectors, especially self-inactivating lentiviral vectors, are considered to be much safer than gamma-retroviral vectors. However self-inactivating lentiviral vectors also have encountered with some unsafe factors and one of them is the problem of transcriptional "read-through" . During the past years, achievements have been made to reduce lentiviral vector transcriptional read-through, which are reviewed herein.
Animals ; Genetic Therapy ; adverse effects ; methods ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Promoter Regions, Genetic ; Proto-Oncogene Proteins ; genetics ; Transcription, Genetic ; genetics ; Virus Inactivation ; Virus Integration