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PubMed2002

Extracts of Ascaris suum egg and adult worm share similar immunosuppressive properties.

Adult Ascaris suum body extract (Asc) prepared from male and female worms (with stored eggs) down-regulates the specific immune response of DBA/2 mice to ovalbumin (OA) and preferentially stimulates a Th2 response to its own components, which is responsible for the suppression of the OA-specific Th1 response. Here, we investigated the participation of soluble extracts prepared from male or female worms or from eggs (E-Asc) in these immunological events. Extracts from either sex (1 mg/animal) or E-Asc (0.35 or 1 mg protein/animal) suppressed the delayed-type hypersensitivity (DTH) reaction (60-85%), proliferative response (50-70%), IL-2 and IFN-gamma secretion (below detection threshold) and IgG1 antibody production (70-90%) of DBA/2 mice to OA. A dose of 0.1 mg E-Asc/animal did not change DTH or proliferation, but was as effective as 0.35 mg in suppressing IL-2 and IFN-gamma, and OA-specific IgG1 antibodies. Lymph node cells from DBA/2 mice injected with Asc (1 mg/animal) or a high dose of E-Asc (1 mg protein/animal) secreted IL-4 upon in vitro stimulation with concanavalin A. As previously demonstrated for Asc, the cytokine profile obtained with the E-Asc was dose dependent and changed towards Th1 when a low dose (0.1 mg protein/animal) was used. Taken together, these results suggest that adult worms of either sex and eggs induce the same type of T cell response and share similar immunosuppressive properties.

Animals↗

PubMed2006

CpG methylation profiles of endothelial cell-specific gene promoter regions in adipose tissue stem cells suggest limited differentiation potential toward the endothelial cell lineage.

In vivo endothelial commitment of adipose stem cells (ASCs) has scarcely been reported, and controversy remains on the contribution of ASCs to vascularization. We address the epigenetic commitment of ASCs to the endothelial lineage. We report a bisulfite sequencing analysis of CpG methylation in the promoters of two endothelial-cell-specific genes, CD31 and CD144, in freshly isolated and in cultures of ASCs before and after induction of endothelial differentiation. In contrast to adipose tissue-derived endothelial (CD31(+)) cells, freshly isolated ASCs display a heavily methylated CD31 promoter and a mosaically methylated CD144 promoter despite basal transcription of both genes. Methylation state of both promoters remains globally stable upon culture. Endothelial stimulation of ASCs in methylcellulose elicits phenotypic changes, marginal upregulation of CD31, and CD144 expression and restrictive induction of a CD31(+)CD144(+) immunophenotype. These events are accompanied by discrete changes in CpG methylation in CD31 and CD144 promoters; however, no global demethylation that marks CD31(+) cells and human umbilical vein endothelial cells occurs. Immunoselection of CD31(+) cells after endothelial stimulation reveals consistent demethylation of one CpG immediately 3' of the transcription start site of the CD31 promoter. Adipogenic or osteogenic differentiation maintains CD31 and CD144 methylation patterns of undifferentiated cells. Methylation profiles of CD31 and CD144 promoters suggest a limited commitment of ASCs to the endothelial lineage. This contrasts with the reported hypomethylation of adipogenic promoters, which reflects a propensity of ASCs toward adipogenic differentiation. Analysis of CpG methylation at lineage-specific promoters provides a robust assessment of epigenetic commitment of stem cells to a specific lineage.

Adipose Tissue↗

PubMed1997

Subjective symptomatology of asthma: validation of the asthma symptom checklist in an outpatient Spanish population.

The objective of this study was to validate the Asthma Symptom Checklist (ASC) so that it could be reliably used to assess the subjective symptomatology of asthma attacks in our context. Subjective symptomatology of asthma was examined in a group of 100 adult Spanish outpatients (57 women, 43 men; 17-69 years of age) with asthma. All of them completed the modified version of the ASC as well as questionnaires of depression, anxiety, and self-management of asthma (self-efficacy expectancies and health care utilization). Data about duration and severity of asthma, as well as dyspnea and %FEV1, were also recorded. The highest reliability Cronbach alpha indexes were for the panic-fear and fatigue scales. The oblique rotation of the ASC revealed five correlated factors (53% of the total variance explained): 1) panic-fear, 2) airways obstruction, 3) airways obstruction and panic-fear, 4) fatigue and irritability, 5) hyperventilation. The structure of factors was revalidated using orthogonal (varimax) rotation. Construct validity was examined by Person product-moment coefficient correlations, ANOVAs (asthma severity x ASC scores), and t-tests (sex by ASC scores). Panic-fear showed the best construct validity, as it was related to the severity of the asthma and the use of high-cost health care resources. There were no differences in ASC scores either on the basis of the asthma severity or on the sex of patients. The ASC factors represent stable components of subjective symptomatology of asthma attacks, especially with regard to the panic-fear and the hyperventilation subscales; however, the structure of the checklist as a whole was not identical to those reported in other studies. Correlations of the ASC with clinical variables related to asthma severity support the construct validity of the instrument and confirm its utility to evaluate the subjective symptomatology of asthma attacks in outpatients.

Adolescent↗

PubMed2004

Effects of cetylpyridinium chloride, acidified sodium chlorite, and potassium sorbate on populations of Escherichia coli O157:H7, Listeria monocytogenes, and Staphylococcus aureus on fresh beef.

The effects of selected food-grade antimicrobial agents at decreasing the number of pathogenic bacteria on fresh beef were determined. Beef cubes inoculated with Escherichia coli O157:H7, Listeria monocytogenes, or Staphylococcus aureus were sprayed with 0.5% cetylpyridinium chloride (CPC), 0.12% acidified sodium chlorite (ASC), 0.1% potassium sorbate (PS), or an equal mix of any two solutions. The beef samples were placed on absorbent tray pads sprayed with each single or mixed solution, wrapped with polyvinyl chloride film, heat sealed, and stored at 4 degrees C for 2 weeks. Surface sanitization using CPC, ASC, or an equal mix of these two agents effectively reduced microbial numbers on the beef during storage. At day 0, ASC and the CPC-ASC mix reduced the number of E. coli O157:H7 by 2.50 and 1.58 log CFU/cm2, respectively. CPC demonstrated a 3.25-log reduction of L. monocytogenes and a 4.70-log reduction of S. aureus at 14 days. The CPC-PS mix reduced E. coli O157:H7 numbers by 1.46, L. monocytogenes by 2.95, and S. aureus by 4.41 log CFU/cm2 at 14 days. PS alone and the mixed solutions, CPC-ASC, CPC-PS, or ASC-PS, were not as effective as ASC or CPC alone. To effectively reduce E. coli O157:H7, L. monocytogenes, or S. aureus numbers, higher (> 0.1%) concentrations of PS were necessary. Loss of redness and light color of beef surfaces consistently coincided with decreases in pH for ASC-treated beef samples.

Animals↗

PubMed2006

Acidified sodium chlorite treatment for inhibition of Listeria monocytogenes growth on the surface of cooked roast beef.

The effects of acidified sodium chlorite (ASC) against Listeria monocytogenes on the surface of cooked roast beef were investigated. L. monocytogenes, strain V7, serotype 1/2a, was inoculated at numbers of 6.0 log CFU/g onto 5-g cubes of cooked regular or spicy roast beef. The samples were allowed to air dry for 1 h. The cooked roast beef samples were dipped into ASC or sprayed with ASC solutions of 250, 500, 750, or 1,000 ppm, then placed in bags with or without a vacuum and refrigerated at 4 degrees C. L. monocytogenes counts were determined after 0, 7, 14, 21, and 28 days of storage by spread plating roast beef samples onto Oxford agar plates that were incubated at 37 degrees C for 48 h. At day 28, the number of L. monocytogenes on the > or = 500 ppm ASC-treated spicy roast beef samples had count reductions that were >4.0 log CFU/g, whereas the same concentrations of ASC-treated regular roast beef samples had approximately a 2.5 log CFU/g reduction in L. monocytogenes counts when compared with the untreated samples. No significant differences (P > 0.05) were observed in L. monocytogenes counts between the vacuum- or nonvacuum-packaged ASC-treated cooked roast beef samples. Sensory evaluation showed no significant differences (P > 0.05) between ASC-treated and untreated roast beef. ASC can be used as a processing aid in the form of a dip or spray treatment to control L. monocytogenes on the surface of cooked roast beef.

Animals↗

PubMed2000

The effects of recombinant RA538 and antisense c-myc adenovirus on tumor cells and the molecular mechanism concerned.

OBJECTIVE: Compare the biological effects of recombinant RA538 and antisense c-myc adenovirus on human gastric cancer cell line (SGC7901) and explore the molecular mechanism in vitro and in vivo. METHODS: SGC7901 cells were treated with Ad-RA538, Ad-AS c-myc or Ad-LacZ. MTT, DNA ladder, TUNEL and FCM, RT-PCR, Western blot analysis, the tumorigenicity in nude mice and experimental therapy of the nude mice were used. RESULTS: Ad-RA538 and Ad-ASc-myc could strongly inhibit cell growth and induce apoptosis of SGC7901 cells. The growth of the Ad-RA538- and Ad-ASc-myc-infected SGC7901 cells were inhibited by 76.3% and 44.1% respectively. The over expression of RA538 and ASc-myc could down-regulate expression of c-myc, bcl-2 and cyclinD1 gene and up-regulate expression of bax gene, but it could not regulate expression of p53, p16, TGase, and ras gene. The tumorigenicity of Ad-RA538 or Ad-ASc-myc in nude mice showed that three of three mice failed to form tumor, compared with Ad-LacZ and parent SGC7901 cells from 7 to 30 days. Experimental therapy of the nude mice bearing subcutaneous tumor of SGC7901 cells showed that intratumor instillation of Ad-RA538 and Ad-ASc-myc inhibited the growth of the tumors. Ad-RA538- and Ad-ASc-myc-treated tumors were inhibited by 60.7% or 68.9% respectively, compared with the tumor injected with Ad-LacZ and mock. CONCLUSION: The expression of Ad-RA538 and Ad-ASc-myc can inhibit growth and induce apoptosis of gastric cancer cell in vitro and in vivo. RA538 and ASc-myc relate to c-myc, bcl-2, cyclinD1 and bax gene closely and have noticeable biologic effects on gastric cancer cells.

Adenoviridae↗

PubMed2006

[Evaluation of clinical management strategies for atypical squamous cells of undetermined significance in cervical cytology].

OBJECTIVE: To evaluate the 3 different clinical management strategies for patients with cervical cytological atypical squamous cells of undetermined significance (ASC-US) recommended by the guideline of The 2001 Bethesda system and the 2001 American Society for Colposcopy and Cervical Pathology (ASCCP). METHODS: 1394 patients with a cytopathological diagnosis of ASC-US by use of liquid-based thin-layer preparation were managed by three different clinical strategies, and evaluated by the percentage of histological diagnosis > or = high-grade intraepithelial lesion (HSIL), i.e., cervical intraepithelial neoplasia (CIN2 and 3) as standard. 421 patients in Group A underwent colposcopically directed cervical biopsy, 475 patients in Group B were followed-up after 6 months by cytology, colposcopy and biopsy were performed if the results were > or = ASC-H or ASC-US and HPV-DNA (+). 498 patients in Group C: underwent HC-II test, colposcopy and biopsy were performed on those aged > or = 30 and with the HPV-DNA (+), if the patients were aged < 30 and with the HPV-DNA (+), HC-II test and cytology would be performed after 6 months; colposcopy and biopsy were performed on those with the results > or = ASC-H or HPV-DNA (+). RESULTS: (1) The results of histological diagnosis > or = CIN2 were found in 27 cases (6.41%) of Group A, 26 cases (5.78%) of Group B, and 34 cases (6.91%) of Group C. There was no statistically significant difference among these 3 groups (all P > 0.05). (2) Convenience was significantly different among these 3 groups (P < 0.01). The workloads for the doctors and the discomfort resulting from biopsy for the patients were the greatest in Group A. The patient's compliance of Group B was low because of the necessity to wait for follow-up six months later. The cost of Group C was relatively higher. (3) In Group C, 66% of the ASC-US patients with HPV-DNA (+) were aged > or = 30. The percentage of histological diagnosis > or = CIN2 was 5.69% in those aged > or = 30 and was 1.22% in those aged < 30. CONCLUSION: Both protocols B and C are practical for clinical management of ASC-US in China. HPV infection is the necessary cause of cervical carcinoma, so the protocol C (cytology and HC-II test) is better for cervical lesion screening. The ASC-US patients aged > or = 30 and with HPV-DNA (+) are at high risk.

Adult↗

PubMed1991

[Effect of L-ascorbic acid 2-phosphate on cultured rabbit keratocytes (the second report)].

We examined the effect of L-ascorbic acid 2-phosphate (P-Asc) on the proliferation of cultured rabbit keratocytes. P-Asc is a derivative of L-ascorbic acid and it yields more prolonged effects of vitamin C in solution than L-ascorbic acid. The proliferation of cultured rabbit keratocytes was promoted by the presence of P-Asc in culture medium for 10, 20, and 30 days. Transmission electron microscopic observations revealed that the cells were more multilayered in the presence of P-Asc (0.1mM) for thirty days than those in the absence of P-Asc. Moreover, this effect of P-Asc was attenuated by azetidine 2-carboxylic acid which is an inhibitor of collagen synthesis. Hence, it is suggested that the promotive effect of P-Asc on the growth of cultured keratocytes is related to the synthesis of collagen. Based on our observations, P-Asc may have a therapeutic effect on corneal stromal damages such as a corneal chemical burn and surgical trauma.

Animals↗

PubMed1996

Detection of an ascorbate radical in an irradiated mice using electron spin resonance (ESR).

Ascorbate radical (Asc.-) produced by the reaction of AscH- (ascorbic acid) with HO or O2.- after irradiation in mice has been measured. It is possible to measure Asc.- easily using ESR and a dialysis method in which Asc.- is collected at room temperature in the interstitial fluid through the dialysis membrane. After irradiation, Asc.- increases in both normal muscle and tumor tissues (SCC-VII) in proportion to the radiation dose. These results suggest that the amount of HO and O2.- produced is reflected in the Asc.- production. This method has been found more useful for the following reasons, (i) no special treatment, such as freezing of the sample, and no administration of noxious agents are necessary to measure Asc.-, (ii) irradiation using a dose of only a few Gy shows an increase in production of Asc.-, and (iii) this method dose not require removal of organs. Using this method, Asc.- can serve as an indicator of the amount of HO and O2.- produced by irradiation in vivo.

Animals↗

PubMed1998

Effects of polyelectrolyte complex (PEC) on human periodontal ligament fibroblast (HPLF) function. II. Enhancement of HPLF differentiation and aggregation on PEC by L-ascorbic acid and dexamethasone.

In addition to many types of extra cellular matrix (ECM) in vivo, cells are stimulated by many types of vitamins, hormones, growth factors, etc. In this paper the effects of L-ascorbic acid 2-phosphate (Asc-2P) and dexamethasone (Dex) on proliferation and differentiation of human periodontal ligament fibroblast (HPLF) using polyelectrolyte complex (PEC) as a matrix in vitro will be discussed. The PEC was composed of chitosan as a polycation, with carboxymethyl (CPEC) or sulfated chitin (SPEC). Asc-2P (0.2 mM) inhibited the growth of HPLF on CPEC, but promoted the growth on SPEC. Moreover, the aggregation of HPLF on CPEC was inhibited by Asc-2P, but that on SPEC was induced in the presence of Asc-2P and Dex. Although Asc-2P reduced an increase in alkaline phosphatase (ALPase) activity of HPLF on CPEC as well, it induced a twofold increase in ALPase activities on SPEC and TCD. Furthermore, in the medium containing Asc-2P and 100 mM of Dex, cell growth was inhibited, but ALPase activity was promoted on both SPEC and TCD to form many aggregates on SPEC. ALPase activity increased by twofold over that of HPLF cultured in the medium containing only Asc-2P. Therefore, it is suggested that the cell functions of HPLF are controlled by the combination of PEC and additives.

Alkaline Phosphatase↗

PubMed2001

Delayed and deficient establishment of the long-term bone marrow plasma cell pool during early life.

Early life antibody responses are characterized by a rapid decline, such that antigen-specific IgG antibodies decline to baseline levels within months following infant immunization. This generic observation remains unexplained. Here, we have analyzed the induction and organ-localization of antigen-specific IgG antibody-secreting cells (ASC) following immunization of 1-week-old or adult BALB/c mice with tetanus toxoid (TT), a T-dependent antigen. Early life priming induced only slightly lower numbers of TT-specific IgG ASC in the spleen, and these reached adult levels following repeat immunization. In contrast, early life immunization generated much fewer bone marrow plasma cells than in adults, even after boosting. A similar limitation of the natural development of the bone marrow pool of ASC was observed. Transfer experiments with adult or early life spleen ASC indicated limited homing of TT-specific adult ASC to the bone marrow of 4-week-old mice as compared to adult recipients, whereas homing patterns were similar when early life or adult ASC were transferred into adult recipients. These observations suggest that a limited bone marrow B cell homing capacity and, as a result, relatively deficient bone marrow ASC responses, are critical factors which may explain the limited persistence of IgG antibodies to T-dependent antigens in early life.

Adoptive Transfer↗

PubMed2006

Clinical implications of the diagnosis "atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion" in pregnant women.

BACKGROUND: Atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H) has a high predictive value for high-grade intraepithelial lesion (HSIL) in the general population. However, the significance of ASC-H in pregnant women remains to be elucidated. The objective of this study was to investigate the clinical implications and pathologic significance of ASC-H in pregnant women, so that these patients will be managed appropriately. METHODS: All Papanicolaou tests that were diagnosed as ASC-H in pregnant women over 1.5 years (total, 60 women) were reviewed and correlated with histologic and/or cytologic follow-up. High-risk type of human papillomavirus (HPV) status was also correlated with follow-up findings. The following cytomorphologic parameters were evaluated for each woman and were compared between the squamous intraepithelial lesion (SIL) follow-up group and the benign follow-up group: inflammatory background, the number of atypical cells, cell arrangement pattern, nuclear irregularity/grooves, hyperchromasia, and cell shape. RESULTS: Among 30 women who had histologic follow-up, 3 women (10%) had HSIL, and 13 women (43%) had low-grade intraepithelial lesion (LSIL). Among 32 women who had cytologic follow-up, 2 women (6%) had HSIL, 3 women (9%) had LSIL, 1 woman (3%) had ASC-H, and 3 women (9%) had atypical squamous cells of undetermined significance (ASCUS). HPV was detected in 24 of 43 women (56%). The cytomorphologic features were similar in the SIL follow-up group and the benign follow-up group. No specific cytomorphologic features that predicted underlying SIL were identified. CONCLUSIONS: ASC-H in pregnant women had a lower predictive value for an underlying HSIL compared with the general population. A positive HPV test result was not a good indicator for an underlying SIL, but a negative result appeared to be useful for ruling out an underlying HSIL. Because of low positive predictive value for HSIL and the difficult colposcopic examination, a more conservative follow-up may be reasonable for pregnant women who have a diagnosis of ASC-H. HPV testing may be used as an adjunctive test.

Adolescent↗

PubMed2004

Slow-down of age-dependent telomere shortening is executed in human skin keratinocytes by hormesis-like-effects of trace hydrogen peroxide or by anti-oxidative effects of pro-vitamin C in common concurrently with reduction of intracellular oxidative stress.

The cellular life-span of cultivated human skin epidermis keratinocytes NHEK-F was shown to be extended up to 150% of population doubling levels (PDLs) by repetitive addition with two autooxidation-resistant derivatives of ascorbic acid (Asc), Asc-2-O-phosphate (Asc2P), and Asc-2-O-alpha-glucoside (Asc2G), respectively, but to be not extended with Asc itself. In contrast, hydrogen peroxide (H(2)O(2)) as dilute as 20 microM which was non-cytotoxic to the keratinocytes, or at 60 microM being marginally cytotoxic achieved the cellular longevity, unexpectedly, up to 160 and 120% of PDLs, respectively, being regarded as a hormesis-like stimulatory effect. The lifespan-extended cells that were administered with Asc2P, Asc2G, or 20 microM H(2)O(2) were prevented from senescence-induced symptoms such as PDL-dependent enlargement of a cell size of 14.7 microm finally up to 17.4 microm upon Hayflick's limit-called loss of proliferation ability as estimated with a channelizer, and retained young cell morphological aspects such as thick and compact shape and intense attachment to the culture substratum even upon advanced PDLs, whereas other non-extended cells looked like thin or fibrous shape and large size upon lower PDLs. The PDL-dependent shortening of telomeric DNA of 11.5 kb finally down to 9.12-8.10 kb upon Hayflick's limit was observed in common for each additive-given cells, but was decelerated in the following order: 20 microM H(2)O(2) > Asc2P = Asc2G > 60 microM H(2)O(2) > Asc = no additive, being in accord with the order of cell longevity. Intracellular reactive oxygen species (ROS) was diminished by Asc2P, Asc2G or 20 microM H(2)O(2), but not significantly by Asc or 60 microM H(2)O(2) as estimated by fluorometry using the redox indicator dye CDCFH. There was no appreciable difference among NHEK keratinocytes that were administered with or without diverse additives in terms of telomerase activity per cell, which was 1.40 x 10(4)-4.48 x 10(4) times lower for the keratinocytes than for HeLa cells which were examined as the typical tumor cells. Thus longevity of the keratinocytes was suggested to be achieved by slowdown of age-dependent shortening of telomeric DNA rather than by telomerase; telomeres may suffer from less DNA lesions due to the continuous and thorough repression of intracellular ROS, which was realized either by pro-vitamin C such as Asc2P or Asc2G that exerted an antioxidant ability more persistent than Asc itself or by 20 microM H(2)O(2) which diminished intracellular ROS assumedly through a hormesis-like effect.

Ascorbic Acid↗

PubMed2004

Cytoprotection by bcl-2 gene transfer against ischemic liver injuries together with repressed lipid peroxidation and increased ascorbic acid in livers and serum.

The maximum gene exhibition was shown to be achieved at 48 h after transfection with human bcl-2 (hbcl-2) genes built in an SV40 early promoter-based plasmid vector and HVJ-liposome for cultured rat hepatocytes. The similar procedure of hbcl-2 transfection was therefore conducted for livers in rats via the portal vein, and after 48 h followed by post-ischemic reperfusion (I/R) operation for some hepatic lobes. The I/R-induced hepatic injuries were in situ observed as both cell morphological degeneration and cellular DNA strand cleavages around capillary vessels of the ischemic liver lobes as detected by HE stain and TUNEL assay, and were biochemically observed as release of two hepatic marker enzymes AST and ALT into serum. All the I/R-induced injuries examined were appreciably repressed for rats transfected with hbcl-2; hbcl-2 was expressed in hepatocytes around the capillaries of ischemic regions such as the median lobe and the left lobe, but scarcely around those of non-ischemic regions. Thus cytoprotection against I/R-induced injuries may be attributed to the I/R-promoted expression of transferred hbcl-2 genes. The possibility was examined firstly by methylphenylindole method, which showed that I/R-enhanced lipid peroxidation in the reference vector-transfected livers were markedly repressed in the hbcl-2-transfected livers. Contents of ascorbic acid (Asc) in serum and livers of hbcl-2-transfected rats were enriched, unexpectedly, versus those of non-transfected rats, and were as abundant as 1.90-fold and 1.95- to 2.60-fold versus those in the pre-ischemic state, respectively. After I/R, an immediate decline in serum Asc occurred in hbcl-2-transfectants, and was followed by prompt restoration up to the pre-ischemic Asc levels in contrast to the unaltered lower Asc levels in non-transfectants except a transient delayed increase. Hepatic Asc contents were also diminished appreciably at the initial stage after I/R in the ischemic lobes of hbcl-2-transfectants, which however retained more abundant Asc versus non-transfectants especially at the initial I/R stage when scavenging of the oxidative stress should be most necessary for cytoprotection. The results showed a close correlation between cytoprotection by exogenously transferred hbcl-2 and repressive effects on the lipid peroxidation associated with Asc consumption or redistribution.

Animals↗

PubMed2004

Cytoprotection of vascular endotheliocytes by phosphorylated ascorbate through suppression of oxidative stress that is generated immediately after post-anoxic reoxygenation or with alkylhydroperoxides.

Vascular endotheliocytes BAE-2 underwent the gradually proceeding cell death until 48 h after reoxygenation (Reox) following 3 h anoxia (Anox), but protected by pre-Anox administration with L-ascorbic acid (Asc)-2-O-phosphate (Asc2P), an autooxidation-resistant Asc derivative, but not by Asc itself. This cytoprotection with Asc2P was achieved in a glucose (Glc)-lacking buffer more advantageously than in a Glc-containing buffer where less efficiency had been demonstrated for Asc entry into BAE-2 cells than in a Glc-lacking buffer. Superoxide anion radicals were detected explosively in the extracellular space at 2-5 min after Reox following the Anox treatment of HUVE endotheliocytes, and were thereafter retained at levels as high as approximately one-half of the maximum level until 60 min after Reox, as shown by cytochrome c reduction assay. Superoxide anions at 3 and 60 min after Reox were suppressed by pre-Anox administration with Asc2P, but not with Asc or dehydro-Asc, and were not suppressed by post-Anox administration with Asc2P; the cytoprotection may need the intracellular accumulation of the ROS-scavenging effector Asc that is converted from Asc2P until 3 min after Reox. The ROS-generator tert-butylhydroperoxide (t-BuOOH) also induced both the diminished cell viability and nuclear DNA strand cleavages of BAE-2 endotheliocytes, which were also protected dose-dependently with Asc2P. The cytoprotection was attributed to reduction of intracellular ROS including hydroperoxide and hydrogen peroxide with Asc2P as shown by fluorometry with the redox indicator CDCFH-DA. Thus Anox/Reox-induced cell death can be prevented by Asc2P that suppresses ROS-generation immediately after Reox following Anox more efficiently in the intracellular sphere rather than in the extracellular space.

Animals↗

PubMed1995

Immunization with Ascaris suum extract impairs T cell functions in mice.

The effect of an Ascaris suum extract (Asc) on several T cell functions was studied in mice immunized with Asc and ovalbumin (OA) in complete Freund's adjuvant. Delayed-type hypersensitivity reactions following challenge with aggregated OA were markedly diminished in mice injected with OA plus a high dose of Asc compared to OA-immunized animals. Proliferation and IL-2, IFN-gamma, IL-4, and IL-10 production in OA-stimulated lymph node cells from the OA + Asc-immunized group were also inhibited. Titration of anti-OA antibodies also showed suppression of IgG1, IgG2a, and IgE isotypes in the animals injected simultaneously with the extract. The degree of suppression induced by Asc on OA-specific cell-mediated responses was dose dependent. The profile of cytokines synthesized in response to Asc also changed depending on the injected dose. IL-4 and IL-10 were mainly produced in response to high doses, whereas IL-2 and IFN-gamma were greatly enhanced at a low dose of Asc. These findings indicate that the A. suum extract may impair crucial T cell functions for cell-mediated as well as humoral immune responses to other antigens through the induction of a predominantly Th2-like response.

Animals↗

PubMed1996

A novel recognition site on laminin for the alpha 3 beta 1 integrin.

Synthetic peptide GD-2 is a sequence of amino acids derived from the carboxy-terminal long arm of the A chain of laminin. Previous studies have shown that peptide GD-2 promotes the adhesion of human squamous cell carcinoma (SCC) cells as well as a variety of other cell lines. In this study, we attempted to identify the receptor that SCC cells use to adhere to peptide GD-2. Monoclonal antibodies (mAbs) against a human SCC cell line were generated. One of these mAbs, ASC-1, bound to the surface of SCC cells as determined by flow cytometry. This mAb inhibited SCC cell adhesion to peptide GD-2 and laminin, but not fibronectin or type IV collagen, suggesting that mAb ASC-1 binds to the SCC receptor for the peptide GD-2 sequence of laminin. MAb ASC-1 immunoprecipitated a complex composed of two components of 135 and 116 kDa. Immunoadsorption of ASC-1-binding material from the SCC cell extract by incubation with mAb ASC-1 resulted in the removal of the alpha 3 beta 1 integrin from the extract. Immunohistochemical staining of tissue from a normal human tongue and from a patient with SCC of the tongue revealed that mAb ASC-1 stained the surface of epithelial cells that were in contact with the basement membrane, as well as those cells located two to three layers above the basement membrane. This mAb also stained blood vessels in the squamous tissue. This staining pattern was identical to that observed when the same tissues were stained by a mAb against the alpha 3 integrin subunit. In summary, by use of a new mAb, ASC-1, that recognizes the alpha 3 beta 1 integrin, we have determined that the alpha 3 beta 1 integrin mediates SCC cell adhesion to the peptide GD-2 sequence within laminin.

Amino Acid Sequence↗

PubMed1991

L-ascorbic acid 2-phosphate, a phosphate derivative of L-ascorbic acid, enhances the growth of cultured rabbit keratocytes.

We examined the effect of L-ascorbic acid 2-phosphate (P-Asc) on the proliferation of cultured rabbit keratocytes. P-Asc is a phosphate derivative of L-ascorbic acid and has more prolonged vitamin C activity in solution than does L-ascorbic acid. The proliferation of cultured keratocytes was promoted by the presence of P-Asc in culture medium. Transmission electron microscopic observations revealed that cells were more multi-layered after culture in the presence of P-Asc (0.1 mM) for 30 days than were those cultured in the absence of P-Asc. The effect of P-Asc was abrogated by L-azetidine 2-carboxylic acid, which is an analogue of proline that inhibits the production and secretion of collagen. Our observations support a therapeutic role for P-Asc in the repair of corneal stromal damage such as that caused by a corneal chemical burn.

Animals↗