Biomedical subjects

W Jia

Publications and source records attributed to W Jia.

At least 37 records · Page 2Linked to original sources

PubMed2000

Preparation and characterization of mabs against different epitopes of CD226 (PTA1).

Recently the platelet and T-cell activation antigen 1 (PTA1) was assigned as CD226 at the 7th Conference and Workshop on Human Leukocyte Differentiation antigens (HLDA). PTA1 is mainly expressed on activated T cells, natural killer (NK) cells, platelets and stimulated endotheliocytes, and involved in the differentiation of cytotoxic T lymphocytes (CTL) and NK, as well as platelet activation and aggregation. We raised hybridomas secreting monoclonal antibodies (MAbs) to PTA1 by using the natural PTA1 as immunogen, which was purified from platelets via affinity chromatography. These MAbs, designated FMU1, FMU2, FMU3, FMU4, FMU5, FMU6 and FMU7, could recognize PTA1 cDNA transfected COS7 cells detected by flow cytometry (FCM), and also react with both natural PTA1 and PTA1/Ig fusion protein in indirect enzyme-linked immunoadsorbent assay (ELISA). The biosensor epitope mapping assay showed that the seven MAbs, together with previous PTA1-specific MAbs Leo A1 and New E1, could bind seven distinct epitopes of PTA1, respectively. The panel of MAbs might be new powerful tools to study the structure-function relationship of PTA1 molecule, and to search for the ligand of PTA1.

Animals↗

PubMed2000

[The association between A55V variant in UCP2 gene and body fat distribution, serum lipid profile in Chinese].

OBJECTIVE: Uncoupling protein 2(UCP2) could play an important role in energy metabolism and body weight regulation. The aim of this study was to investigate Ala55Val(A55V) variant in the UCP2 gene has effects on serum lipid profile, body fat and its distribution in Chinese. METHODS: The genotypes of A55V variant in the UCP2 gene were determined by a PCR-RFLP assay in 359 unrelated Chinese [including 193 normal glucose tolerance(NGT) and 166 type 2 diabetic subjects by ADA 97' criteria]. The parameters for regional adipose tissue distribution were measured by magnetic resonance imaging(MRI). RESULTS: In NGT group, an association between A55V variant in the UCP2 gene and body mass index(BMI) (P=0.0246), as well as femoral subcutaneous adipose tissue area (FA) (P=0.0017), was noted in females. A55V variant in the UCP2 gene was also associated with serum triglyceride (TG) level (P=0.0072) in males. However, in type 2 diabetes group, an association between A55V variant in the UCP2 gene and FA (P=0.0150) was replicated in females too. Those females who were homozygotes of AA in the UCP2 gene had decreased FA not only in NGT group but also in type 2 diabetes group. Furthermore, logistic regression analysis indicated that FA (P=0.0098 in NGT females, P=0.0071 in type 2 diabetic females) and BMI (P=0.0016 in NGT females), as well as TG level (P=0.0040 in NGT males) were associated with this variant in the UCP2 gene. CONCLUSION: A55V variant in the UCP2 gene is associated not only with FA (in NGT females and type 2 diabetic females) but also with BMI (in NGT females). Therefore A55V variant in the UCP2 gene appears to play a role in body fat and its distribution in Chinese females.

Adipose Tissue↗

PubMed2000

[Sentinel lymph node biopsy in breast cancer: value for predicting the status of axillary node].

OBJECTIVE: To evaluate feasibility and accuracy of intraoperative lymphatic mapping with sentinel lymphadenectomy (SLND) for predicting axillary status in patients with breast cancer. METHODS: Our study enrolled 52 patients with primary breast cancer, clinically and ultrasonographically negative axillae. Mapping procedures and SLND were performed using methylene blue injected at the primary breast cancer site followed by axillary lymph node dissection (ALND). Sentinel node (SN) was examined by using frozen sections intraoperatively and all of the axillary lymph nodes were evaluated pathologically (HE) after operation. RESULTS: Sentinel nodes were identified in 46 (88.5%) of 52 procedures and nodal status accurately predicted axillary in 44 (95.7%) of 46 cases. In 2 (4.3%) of 46 cases, the SN was false-negative. The overall sensitivity of the SN technique was 90.9%, with a specificity of 100%. The overall positive and negative predictive values were 100% and 92.3%, respectively. In 15 (75%) of 20 cases of clinically negatively and pathologically positive axillary, the SN was the only tumor-involved lymph node identified. CONCLUSION: Our study indicates that intraoperative lymphatic mapping using a vital dye and SLND can accurately predict the axillary status of primary breast cancer patients with clinically and ultrasonographically negative axillae.

Adult↗

PubMed2000

[The early biochemical changes of cataractous lenses of rats cultured in vitro].

OBJECTIVE: To study the mechanism of cataractogenesis. METHODS: Tissue culture was used to study the cataractous lenses of rats induced by sodium selenite or galactose. At the early stage, the content of nonprotein sulfhydryl (NP-SH), protein sulfhydryl (P-SH), non-soluble disulfide bond, malonaldehyde (MDA) and the activities of glutathione peroxidase (GSH-PX), glutathione reductase (GSSG-R) and glutathione-S-transferase (GSH-S) were measured in the cataractous lenses, and they were compared with that of the normal lenses. RESULTS: The two kinds of material could all induce cataract in rats, sodium selenite being more potent. In the early period of culture (lenses were transparent), NP-SH and P-SH were decreased, while disulfide and MDA were increased, the activity of GSH-PX rose obviously, that of GSH-S also had a tendency of rise, however, the activity of GSSG-R had no obvious changes. CONCLUSIONS: Rat lens opacity may occur after the lens is cultured in vitro with the addition of sodium selenite or galactose, and biochemical changes may develop in the lens at the early period of culture (lenses are transparent).

Animals↗

PubMed1999

Investigation on process parameters involved in preparation of poly-DL-lactide-poly(ethylene glycol) microspheres containing Leptospira Interrogans antigens.

Block copolymer, poly-DL-lactide-poly(ethylene glycol) (PELA) with 11.5% of poly(ethylene glycol) (PEG) content was prepared by bulk ring-opening polymerization using stannous chloride as initiator. PELA microspheres with entrapped Leptospira Interrogans antigens, outer membrane protein (OMP) were elaborated by solvent extraction method based on the formation of multiple w/o/w emulsion, and the resulting microspheres were characterized with respect to particle size, OMP entrapment and morphology characteristics. The purpose of the present study is to perform the optimization of preparative parameters for OMP-loaded PELA micropsheres to control particle size and improve the OMP encapsulation efficiency. Of all the parameters investigated, the polymer concentration of organic phase and the external aqueous phase volume play major roles on particle size, while the organic phase volume, internal aqueous phase volume and the addition of surfactant into the internal aqueous phase display considerable effects on OMP loading efficiency. A small volume of internal aqueous phase and intermediate volumes of organic phase and external aqueous phase were favorable to achieve micropsheres with a size of 1-2 microns and high antigen encapsulation efficiency (70-80%). In vitro OMP release profiles from PELA microspheres consist of a small burst release followed by a gradual release phase. The OMP release rate shows some relations with the porous and water-swollen inner structure of the microspheres matrix. The presence of surfactant in microspheres accelerates OMP release, but the OMP entrapment within microspheres shows limited effects on the release profile.

Antigens, Bacterial↗

PubMed1999

Cell surface expression of MHC molecules in glioma cells infected with herpes simplex virus type-1.

9L glioma cells consistently expressed major histocompatibility complex (MHC) class I but not class II molecules. Herpes simplex type-1 virus (HSV-1) infection significantly reduced the expression of MHC I on the cell surface. Recombinant interferons could enhance the cell-surface expression of MHC I but had no effect on MHC II. This enhancement was partially inhibited by HSV-1 infection. HSV-1 mutants with deletions in ICP4, ICP6, ICP27, ICP47 and UL41 genes do not affect the infection induced inhibition, suggest that a different mechanism may be employed in the inhibition of cell-surface expression of MHC molecules.

Animals↗

PubMed1999

Nickel release from orthodontic arch wires and cellular immune response to various nickel concentrations.

AIMS: Results from two previous clinical studies suggested that exposure to high nickel-containing orthodontic arch wires may induce hypersensitivity in certain individuals. The purpose of this study was to measure the amount of nickel released from three types of nickel-containing arch wires into a synthetic saliva in vitro, and determine if the concentrations were sufficient to elicit either cytotoxic (trypan blue exclusion test) or stimulatory (MTT test) responses in human peripheral blood mononuclear cells (PBMCs) derived from nickel-sensitive and nickel-nonsensitive individuals. PBMCs were exposed to five concentrations of nickel sulfate solutions ranging from 0-29 ppm, and results were compared, particularly at concentrations obtained from nickel release experiments. FINDINGS: The amount of nickel released into synthetic saliva ranged from 0.4-4.1 ppb. Wires subjected to a combination of soaking and cyclic straining released significantly more nickel than those that were soaked only (p </= 0.05), and NiTi wires released significantly more nickel than did stainless steel or nitrogen-implanted NiTi wires (p </= 0.05). For PBMCs, significant increased cell proliferation was not observed for any nickel concentration. PBMC cell death rates were highest at nickel concentrations of 29 ppm when the cells were cultured without a cell growth promoter (p </= 0.05), and MTT test values were significantly reduced at both 2.9 and 29 ppm when a growth promoter was included (p </= 0.05). CONCLUSION: The maximum amount of nickel released from all tested arch wires was 700 times lower than the concentrations necessary to elicit cytotoxic reactions in human PBMCs.

Adult↗

PubMed1999

The heterogeneity in the immune response and efficiency of viral dissemination in brain infected with herpes simplex virus type 1 through peripheral or central route.

Using immunohistochemistry on adjacent brain sections, we studied the correlation between the dissemination of the virus, the inflammatory responses and the expression of major histocompatibility complex (MHC) proteins in rat brain infected with herpes simplex virus (HSV-1) F strain by either corneal scarification or intracerebral injection. Our results showed that the mortality of the corneally infected rats was much higher than that of the intracerebrally infected rats, due to a more extensive dissemination of the virus in the brain, particularly in the brain stem. The inflammatory responses were similar in brains infected through either route, as demonstrated by the expression of MHC I/II antigens on infiltrating lymphocytes, leukocytes and macrophage/microglia cells. While there was strong immunoreactivity for HSV-1 antigens in the cerebral cortex, the infiltrates were only located in subcortical areas, especially the hippocampus. Therefore, the distribution of these immune cells did not always overlap with the regions of viral infection. These results suggest that HSV-1 disseminate more efficiently from the peripheral to the central nervous system (CNS) than from CNS to CNS, which is independent of the immune responses, and that the cerebral cortex may immunologically respond to HSV-1 infection differently from other brain regions.

Animals↗

PubMed1999

Ultraviolet radiation-induced color shifts occurring in oil-pigmented maxillofacial elastomers.

STATEMENT OF PROBLEM: Oil-based pigments are added to a maxillofacial prosthesis either as base colorants present within the elastomer or as surface tints that are painted on with an adhesive. Color stability of the pigments and pigmented prosthetic materials on exposure to ultraviolet radiation are unknown. PURPOSE: This study measured DeltaE* color changes caused by ultraviolet radiation for materials colored with 5 oil pigments, applied either as base colorants (intrinsic) or surface tints (extrinsic) to a silicone elastomer. MATERIAL AND METHODS: One of 5 oil pigments was added to polydimethyl siloxane disks to serve as a base colorant (0.2 weight percent present throughout a 2 mm thick disk) or as a concentrated surface tint (2.0 weight percent concentrated in upper 0.3 mm thickness). Pigmented disks, along with pigment-only and elastomer-only control disks, were exposed to ultraviolet radiation for 400, 600 and 1800 hours. DeltaE* color changes were measured at baseline and for each time interval. RESULTS: Control samples underwent minimum color changes after 1800 hours (DeltaE* </= 2.0), whereas samples containing oil pigments as base colorants demonstrated a wide range of susceptibility to ultraviolet radiation, with the greatest changes occurring for pigments cadmium red, cadmium yellow, and yellow ochre (7.1 </= DeltaE* </= 9.4). Elastomers coated with the same oil pigments as concentrated surface tints demonstrated significantly lower color shifting after 1800 hours of radiation exposure (maximum DeltaE* = 4.2, P </= .05). CONCLUSION: Customizing a prosthesis with an oil-pigmented surface tint may reduce the incidence of color change, provided a sufficient amount of pigment is present.

Coloring Agents↗

PubMed1999

Capsid protein-encoding genes of hamster polyomavirus and properties of the viral capsid.

On the basis of its genome organization the hamster polyomavirus (HaPV) is closely related to the murine polyomavirus Py. But HaPV infection, in contrast to Py infection, gives rise to two different tumor types; depending on the hamster strain used for infection, HaPV induces either epitheliomas or lymphomas. Although the HaPV virions were shown to be similar to those of Py and SV40, more precise information about the structure and protein composition of the HaPV capsid was still missing. Here we describe the primary structure of the capsid protein-encoding HaPV genes and the structure and protein composition of the HaPV capsid. Virions isolated from epitheliomas in HaPV-infected hamsters were shown by electron microscopy to be spherical particles with the typical icosahedral structure of polyomaviruses. However, in contrast to the capsids of SV40 and Py, a T = 7 laevo symmetry of HaPV capsids was observed. Separation of HaPV virions in SDS polyacrylamide gels and Western blotting with VP1-specific antisera identified VP1 as the major capsid protein species corresponding in its molecular weight to the predicted value of 41.8 kDa. Because of the presence of two potential translational initiation sites in the VP1 gene, the N-terminal amino acid sequence of virion VP1 was determined and found to start at the second initiation site. The amino acid homologies of HaPV capsid proteins shared with Py varied between 65.5% (VP1), 45.4% (VP3) and 44.6% (VP2), whereas the homologies to the relevant proteins of other polyomaviruses were found to range between 49.6-57.9% for VP1 and 28.9-41% for VP2/VP3.

Amino Acid Sequence↗

PubMed1999

Immunohistochemical localization of dopamine-beta-hydroxylase in human and monkey eyes.

PURPOSE: To investigate the pattern of dopamine-beta-hydroxylase (DBH)-containing fibers in human and monkey eyes. METHODS: DBH-containing fibers were detected by immunohistochemistry. The primary antibody used recognized DBH, the key enzyme in the conversion of dopamine to noradrenaline. RESULTS: In the anterior segment, DBH immunoreaction product was found in the peripheral corneal endothelium layer, in both the dilator and sphincter muscles of the iris, as well as in the anterior border layer of the iris. The ciliary muscle and the stroma of the ciliary processes were also zones of concentration. In the posterior segment, staining was seen around blood vessels in the choroid, in the vascular walls of the short posterior ciliary arteries and in the ciliary nerves. The retina was also immunopositive, with specific labeling in cones and rods of photoreceptors, inner and outer plexiform layers and ganglion cell layer. There was no significant difference in the distribution of DBH-related immunoreactivity in human and monkey eyes. CONCLUSIONS: The localization of DBH-related immunoreactivity is generally consistent with the known physiological roles of noradrenaline. However, an apparently high concentration of the enzyme in the anterior border layer of the iris and in retinal photoreceptors raises questions about the possible role of DBH-containing fibers in these structures.

Adolescent↗

PubMed1999

Isolation of cDNAs encoding gibbon and monkey platelet and T cell activation antigen 1 (PTA1).

Human platelet and T cell activation antigen 1 (PTA1) is a 67kDa type I transmembrane glycoprotein mainly expressed on the surface of activated T cells and platelets, and is involved in the development of human cytotoxic T cell (CTL) as well as platelet activation and aggregation. We have cloned and sequenced gibbon PTA1 (gPTA1) and monkey PTA1 (mPTA1) cDNAs by RT-PCR from gibbon leukemic cell line MLA 144 and PHA-induced Rhesus monkey PBMC respectively. The mature proteins of gPTA1, mPTA1 and human PTA1 (hPTA1) share 93-95% amino acid similarity with the highest similarity in domain 1 of extracellular region. All the important features of PTA1 molecule are conserved among these Primates: (1) the ORF encoding 336 amino acid residues including signal sequence (18aa), extracellular region (232aa), transmembrane sequence (25aa) and cytoplasmic region (61aa); (2) two conserved pairs of Cys (Cys19 to Cys90 and Cys134 to Cys204) forming disulfide bonds stabilizing the two immunoglobulin superfamily V-like domains; (3) eight putative N-linked glycosylation sites (except gPTA1 with nine sites) and three O-linked glycosylation sites in extracellular region; and (4) predicated protein kinase C phosphorylation sites (Thr275 and Ser311), casein kinase II sites (Ser295 and The299) and the potential tyrosine phosphorylation site (Tyr304). These data indicate that PTA1 molecule is highly conserved among the Primates and may play important roles in immune response.

Amino Acid Sequence↗

PubMed1999

Experimental gene therapy for an in vitro model of proliferative vitreoretinopathy.

BACKGROUND: Proliferative vitreoretinopathy (PVR) is the leading cause of failure of retinal reattachment surgery. Since a key component of PVR is cell proliferation, we performed a study to examine whether the ribonucleotide-reductase-deficient herpes simplex virus type I (HSV-I) mutant hrR3 can be effective at destroying proliferating retinal pigment epithelial (RPE) cells and thus prevent epiretinal membrane formation and PVR, while sparing nondividing cells, such as neurons. METHODS: Primary cultures of rat RPE cells and rat cortical neurons were infected with 300 microL of hrR3 HSV-I to achieve a multiplicity of infection of 1.0. After 1 hour at 37 degrees C, 700 microL of growth medium was added to raise the total volume of medium to 1 mL. At 0, 12, 24 and 36 hours the cultures were observed, and the ratio of dead cells to live cells was determined. HSV infection and protein expression were confirmed by a beta-galactosidase histochemical assay or an antihuman HSV-I immunoassay, or both. RESULTS: At 24 hours more than 95% of the RPE cells and neurons stained positively for HSV infection, although beta-galactosidase was expressed predominantly in RPE cells. At 36 hours 72% (standard deviation 2.1%) of the RPE cells were dead. There was no noticeable cell death in the neuronal or mock-infected control cultures. INTERPRETATION: The results suggest that the hrR3 mutant strain of HSV-I can be used to infect and selectively kill actively proliferating rat RPE cells while sparing normal, nonreplicating cells. This model may be used to explore potential therapies for PVR in humans.

Animals↗

PubMed1999

Urinary cross-linked N-telopeptides of type I collagen and bone metabolic diseases.

OBJECTIVE: To evaluate the type I collagen cross-linked N-telopeptide (NTx) levels in human urine as an indicator of bone resorption rate in different ages, sex and in bone metabolic diseases. METHODS: Urinary NTx was determined by immunoassay in 591 Beijing healthy subjects aged from 0 to 86 years and 379 patients with bone metabolic diseases. RESULTS: The levels of urinary NTx were significantly higher in children than in adults (P < 0.001) and higher in boys than in girls (P < 0.01) and increased 1.4-2.2 times in postmenopausal females than in men and premenopausal women. Urine NTx had a positive linear correlation with urine HOP/Cr (r = 0.778, P < 0.01) and Ca/Cr ratio (r = 0.320, P < 0.01), and a negative linear correlation with age (r = -0.523, P < 0.01) and lumbar spine BMD (r = -0.426, P < 0.01). The levels of urine NTx increased for 3.6 times in pregnancy, 1.5 times in osteoporosis, 1.9 times in fragility fracture, 3.6 times in chronic renal failure, 2.1 times in rickets and 7.2 times in multiple myeloma compared to age matched controls. CONCLUSIONS: NTx in urine is a specific and sensitive indicator of bone resorption and is able to distinguish normal premenopause from late osteoporotic patients. NTx could be used as diagnostic information about metabolic bone diseases, and to monitor antiresorptive therapy.

Adolescent↗

PubMed1999

The impact of codon 54 variation in intestinal fatty acid binding protein gene on the pathogenesis of diabetes mellitus in Chinese.

OBJECTIVE: To investigate whether or not the intestinal fatty acid binding protein gene (FABP2)-Ala54Thr variation is related to non-insulin dependent diabetes mellitus (NIDDM), obesity, dyslipidemia and glucose stimulated insulin secretion (GSIS) in Chinese. METHODS: The FABP2-Ala54Thr variation was detected by PCR/Hhal digestion in 231 Chinese subjects (116 with normal glucose tolerance (NGT), 54 with impaired glucose tolerance (IGT) and 61 with NIDDM). Plasma glucose, insulin and C-peptide levels before and after 75 g glucose load as well as fasting lipid profile were determined. RESULTS: (1) The Ala54 and Thr54 allele frequencies in Chinese were 0.71 and 0.29 respectively; (2) The FABP2-Ala54Thr variation was neither associated with fasting and post-challenged plasma glucose levels nor with NIDDM; (3) This variation was neither associated with fasting lipid profile nor with obesity; (4) The IGT subjects with genotype Thr54(+) (Thr54 homozygotes and heterozygotes) had lower fasting, 2-hour and total C-peptide levels and smaller AUC representing lesser C-peptide secretion after glucose challenge than those with genotype Thr54(-) (Ala54 homozygotes) (P = 0.04, 0.03, 0.01 and 0.01 respectively). The serum insulin levels changed in the same tendency. CONCLUSIONS: The glucose stimulated insulin secretion (GSIS) reserve of islet beta-cells is more limited in subjects with FABP2-Thr54(+) genotype than in those with FABP2-Thr54(-) genotype. It suggests that FABP2-codon 54 variation might contribute to the insufficient insulin secretion in the development of NIDDM in Chinese.

Alanine↗

PubMed1999

[Association of Gln223Arg variant in leptin receptor gene with metabolic abnormalities and hypertension in type II diabetes mellitus in Shanghai "Han" population].

OBJECTIVE: To investigate the association of Gln223Arg variant in leptin receptor gene with metabolic abnormalities and hypertension in type II diabetes mellitus. METHODS: The genotypes of Gln223Arg variant in leptin receptor gene were determined by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) assay in 359 unrelated subjects of Shanghai "Han" population (including 193 subjects with normal glucose tolerance and 166 type II diabetic patients). The clinical data were also analyzed. The parameters for regional adipose tissue distribution were measured by magnetic resonance imaging (MRI). RESULTS: Significant difference of genotype frequency of Gln223Arg variant in leptin receptor gene was observed between hypertensive and normotensive type II diabetic male patients (P = 0.008). "A" allele was associated with increased systolic blood pressure (P = 0.0026) and diastolic blood pressure (P = 0.0084) in type II diabetic male patients. Logistic regression analysis showed that this gene variant was an independent risk factor of type II diabetic male patients accompanied with hypertension (P = 0.0031), and it was associated with elevation of systolic blood pressure (P = 0.0054). The odds ratio of hypertension in type II diabetic male patients with "A" allele was 2.825 (95% CI 1.418, 5.627). CONCLUSION: Gln223Arg variant in leptin receptor gene is associated with hypertension in type II diabetic male patients, especially with elevation of systolic blood pressure.

Adult↗

PubMed1999

[Detection of telomerase activity in prostate needle-biopsy samples].

OBJECTIVE: To detect telomerase activity inprostate needle-biopsy samples and its role in diagnosis and prognosis of prostate cancer. METHODS: 20 tissue samples of prostate cancer tissues and 16 samples of adjacent tissues were obtained by needle-biopsy guided by transrectal B ultrasonography. 16 samples of benign prostatic hyperplasia tissues were obtained by prostatectomy and were confirmed pathologically. Telomerase activity was revealed by modified PCR-based telomeric repeat amplication protocol (TRAP)-silver staining assay. RESULTS: Telomerase activity was revealed in 18 cases of prostate cancer. In tissues adjacent to prostate cancer, telomerase activity was found in 7 of 11 cases of prostatic intraepithelial neoplasia (PIN), and in 2 of 5 cases of BPH. None of tissue samples from patients with BPH showed telomerase activity. CONCLUSIONS: Telomerase activity in prostate needle-biopsy samples might be an useful marker for detecting prostate cancer and biological malignancy.

Adenocarcinoma↗

PubMed1999

[Purification and identification of pili of Vibrio cholera O139].

To provide the base of manufacturing V. cholera O139 subunit vaccine, purification and identification methods of V. cholera O139 Toxin-coregulated pili (TCP) were studied. The results showed the optimal expressing condition of TCP was in AKI or CFA medium, at 30 C for 24-36 h with stationary cultivation. The molecular weight of the TCP subunit protein measured by sodium dodecyl sulfate-poly acrylamide gel electrophoresis was about 20.5 kd. By using mouse anti-TCP monoclone antibody and sheep anti-mouse IgG-HRP antibody, the TCP was confirmed by dot blot immunoassay. Rabbit immune serum of TCP purified from O139 strain agglutinated both O139 biotype and El Tor biotype Vibrio cholera strains. These indicate that TCP may be an effective and protective antigen to conduct the preparation of V. cholera vaccine.

Animals↗