Biomedical subjects

N Huang

Publications and source records attributed to N Huang.

At least 37 records · Page 2Linked to original sources

PubMed2001

Are the dominant and recessive plant disease resistance genes similar? A case study of rice R genes and Xanthomonas oryzae pv. oryzae races.

The resistance of rice to its bacterial blight pathogen Xanthomonas oryzae pv. oryzae (Xoo) has both qualitative and quantitative components that were investigated using three near-isogenic line sets for four resistance (R) genes (Xa4, xa5, xa13, and Xa21) and 12 Xoo races. Our results indicate that these two resistance components of rice plants were associated with the properties of the R genes. The qualitative component of the R genes was reflected by their large effects against corresponding avirulent Xoo races. The quantitative component of the R genes was their residual effects against corresponding virulent races and their epistatic effects, which together could lead to high-level resistance in a race-specific manner. Our results revealed important differences between the different types of R genes. Two R genes, Xa4 and Xa21, showed complete dominance against the avirulent Xoo races and had large residual effects against virulent ones. They acted independently and cumulatively, suggesting they are involved in different pathways of the rice defensive system. The third R gene, xa5, showed partial dominance or additivity to the avirulent Xoo races and had relatively small but significant residual effects against the virulent races. In contrast, xa13 was completely recessive, had no residual effects against the virulent races, and showed more pronounced race specificity. There was a strong interaction leading to increased resistance between xa13 and xa5 and between either of them and Xa4 or Xa21, suggesting their regulatory roles in the rice defensive pathway(s). Our results indicated that high-level and durable resistance to Xoo should be more efficiently achieved by pyramiding different types of R genes.

Genes, Dominant↗

PubMed2001

Amplification effect and mechanism of action of ET-1 in U-46619-induced vasoconstriction in pig skin.

The aim of this study was to investigate if a low concentration of endothelin-1 (ET-1; 8 x 10(-10) M) may amplify the skin vasoconstrictor effect of other vasoactive substances in the pathogenesis of skin vasospasm. Pig skin flaps (6 x 16 cm) were perfused with Krebs buffer equilibrated with 95% O(2) and 5% CO(2) at 37 degrees C and pH 7.4. Skin perfusion pressure measured by a pressure transducer and skin perfusion assessed by the dermofluorometry technique were used for assessment of skin vasoconstriction. We observed that ET-1 (8 x 10(-10) M) significantly amplified the concentration-dependent (10(-7)-10(-5) M) skin vasoconstrictor effect of norepinephrine. More importantly, we observed for the first time that this low concentration of ET-1 also amplified the concentration-dependent (10(-8)-10(-6) M) skin vasoconstrictor effect of the thromboxane A(2) mimetic U-46619, and this amplification effect of ET-1 was completely blocked by the protein kinase C (PKC) inhibitor chelerythrine (5 x 10(-6) M). Conversely, the PKC activator phorbol 12,13-dibutyrate (10(-7) M) amplified the vasoconstrictor effect of U-46619. Furthermore, the sensitivity of the skin vasculature to the vasoconstrictor effect of extracellular Ca(2+) in U-46619-induced skin vasoconstriction was significantly enhanced in the presence of 8 x 10(-10) M ET-1. Finally, the cyclooxygenase inhibitor indomethacin (5 x 10(-6) M) did not affect the amplification effect of ET-1 on U-46619-induced skin vasoconstriction. We conclude that a low concentration of ET-1 can amplify the skin vasoconstrictor effect of U-46619 independent of endogenous cyclooxygenase products, and the mechanism may involve activation of PKC and increase in sensitivity of the contractile apparatus to Ca(2+) in smooth muscle cells.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

PubMed2001

Effect of nicotine on vasoconstrictor and vasodilator responses in human skin vasculature.

Our objective was to test the hypothesis that acute exposure of human skin vasculature to nicotine may have deleterious effects on endothelial function. Vasoconstriction and vasorelaxation in isolated perfused human skin flaps (approximately 8 x 18 cm) derived from dermolipectomy specimens were assessed by studying changes in skin perfusion pressure measured by a pressure transducer, and skin perfusion was assessed by a dermofluorometry technique (n = 4 or 5). It was observed that nicotine (10(-7) M) amplified (P < 0.05) the norepinephrine (NE)-induced concentration-dependent (10(-7)-10(-5) M) increase in skin vasoconstriction compared with the control. This amplification effect of nicotine in NE-induced skin vasoconstriction was not blocked by the nicotine-receptor antagonist hexamethonium (10(-6) M) or the cyclooxygenase inhibitor indomethacin (10(-5) M). It was also observed that ACh and nitroglycerin (NTG) elicited a concentration-dependent (10(-8)-10(-5) M) vasorelaxation in skin flaps preconstricted with 8 x 10(-7) M of NE. The vasorelaxation induced by ACh was attenuated (P < 0.05) in the presence of nicotine (10(-7) M) compared with the control. However, skin vasorelaxation induced by NTG was not affected by nicotine (10(-7) M). ACh and NTG are known to induce endothelium-dependent and -independent vasorelaxation, respectively. The present findings were interpreted to indicate that acute exposure of human skin vasculature to nicotine was associated with 1) amplification of NE-induced skin vasoconstriction and 2) impairment of endothelium-dependent skin vasorelaxation. Cyclooxygenase products and nicotine receptors blocked by hexamethonium were not involved in the amplification of NE-induced skin vasoconstriction by nicotine. These findings may provide further insight into the pathogenesis of skin vasospasm in skin flap surgery and skin ischemic disease associated with cigarette smoking or use of smokeless tobacco.

Acetylcholine↗

PubMed2001

Familial Creutzfeldt-Jakob disease associated with a point mutation at codon 210 of the prion protein gene.

Creutzfeldt-Jakob disease (CJD), the most known human prion disease, is usually sporadic but approximately 15% of the cases are familial. To date, seven CJD cases with codon 210 mutation (GTT to ATT) have been reported in the literature. We describe a case of a 57 year-old woman who presented gait disturbances and rapidly progressive dementia, leading to death four months after onset. Electroencephalogram revealed periodic activity, diffusion-weighted magnetic resonance imaging showed hypersignal in basal ganglia, and test for 14-3-3 protein was strongly positive in the CSF. The complete prion protein gene coding region was sequenced after PCR amplification, showing a point mutation in codon 210. This is the first case of CJD with codon 210 mutation diagnosed in Brazil. We emphasize the role of genetic search for prion protein gene mutation, even in patients presenting clinical features resembling sporadic CJD.

Codon↗

PubMed2001

[Human toll like receptor gene expression in human pulmonary granule epithelial cells and umbilical vein endothelial cells].

OBJECTIVE: To determine whether human Toll like receptors (TLRs) genes are expressed in human lung granule epithelial cells and umbilical vein endothelial cells (UVECs). METHODS: Specific primers for TLR2 and TLR4 were designed and total RNAs were isolated from human monocyte cell line THP-1, human lung granule epithelial cell line SPC-A-1 and the primary culture of human UVECs. RT-PCR and Northern blotting with digoxin labeling were used for the detection of TLR2 and TLR4 mRNA expression. RESULTS: TLR4 mRNA expressed in the three types of cells mentioned above. TLR2 mRNA was detected in THP-1 and UVECs, but not in SPC-A-1 cells. By using DNA sequencing, the RT-PCR products were confirmed to be TLR2 and TLR4 cDNAs. CONCLUSION: This study provided evidence indicating that Toll like receptor genes constitutively express not only in human monocytes but also in human larger vascular endothelial cells and pulmonary granule epithelial cells.

Cells, Cultured↗

PubMed2001

[An immunohistochemical study of distribution of insulin in lateral pterygoid muscle after functional mandibular protrusion in rats].

OBJECTIVE: The aim of this study is to investigate the effects of insulin on the lateral pterygoid muscle of growing rats after the functional mandibular protrusion. METHODS: An immunohistochemical (the ABC method) method was used to detect the location and distribution of insulin in the lateral pterygoid muscle of rats. RESULTS: 1. Insulin located in the cell membrane of the lateral pterygoid muscle. 2. The distribution of insulin in the lateral pterygoid muscle of rats was enhanced after the functional mandibular protrusion. CONCLUSION: Insulin may play an important role in adaptive remodeling of the lateral pterygoid muscle after the treatment of functional mandibular protrusion.

Animals↗

PubMed2001

[Research and establishment of the computerized orthodontic and orthopedic prediction system].

OBJECTIVE: The aim of this study was to develop a computerized orthopedics and orthodontics interactive prediction system and verify its reliability. METHODS: 24 cases of Angel II division I patients were selected in the study. All of them had been treated by activator or Frankel functional appliance, and had the satisfied therapy result. Both pre and post-treatment cephalogram of them were used verifying veracity of therapy prediction. RESULTS: All of cephalometric measure items include 11 angle measure items and 14 distance measure items. There were 9 coincidence indicators in angle measure items and 7 in distance measure items. CONCLUSION: The results indicate it is liable and effective to use the computerized orthopedics and orthodontics system for prediction.

Activator Appliances↗

PubMed2001

[Establishment and research of the computerized profile and photo prediction system].

OBJECTIVE: The study aims to develop a lateral oral facial profile and photo prediction system and then to verify its reliability. METHODS: 24 paired lateral cepholagrams in both pre- and post-treatment were selected and analyzed by three soft tissue cephalometric analysis methods in this study. All cases were of the angel II division I malocclusion, and had been successfully treated with the orthopedics therapy by activator or Frankel appliance. Soft tissue profile prediction had been automatically completed, following the hard tissue interactive orthopedics prediction approach in the pretreatment lateral cephalogram, and the predictive results were compared to the same cepholametric analysis in post-treatment cephalogram. RESULTS: There are 19 coincidence indicators in 30 measurements of three soft tissue cephalometric analysis, and 7 in 9 angel measurements, 10 in 18 instance measurements, 2 in 3 instance ratio measurements is consistency, respectively. CONCLUSION: The results indicate that the lateral oral facial profile and photo prediction system is effective in profile prediction.

Activator Appliances↗

PubMed2001

[K. pneumoniea endotoxin induced mice beta-defensin-4 mRNA expression and its signaling transduction].

OBJECTIVE: To investigate the in vivo effects of Klebsiella pneumoniae endotoxin(LPS) on beta-defensin expression and the relevant signaling transduction pathway. METHODS: A LPS tolerant mouse C3H/HeJ with a point mutation at Toll-like receptor-4 (TLR4) gene and its wild type strain C3H/HeN were used in this study. C3H/HeJ and C3H/HeN were injected with 4 mg/kg of LPS intraperitoneally. The tracheas, lungs and kidneys of the C3H/HeJ and C3H/HeN were collected respectively at different LPS-treated time points, and the total RNA of each sample was extracted. The expression of mice beta-defensin-3 and/or beta-defensin-4 mRNA in these tissues was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The sequence of cDNA amplified from the lung of C3H/HeN treated by LPS for 24 h was analyzed. By using western blot, p-I kappa B alpha (phosphorylated I kappa B alpha) and I kappa B alpha of in the lungs of C3H/HeJ and C3H/HeN were detected at different time points after treatment with LPS or without LPS. RESULTS: 1. beta-defensin-4 mRNA was detected in the lungs of C3H/HeN after 24 h treatment with LPS. In contrast, no signal was determined in C3H/HeJ mice with LPS treatment and the C3H/HeN mice without LPS treatment. 2. Compared with the control, increas of the p-I kappa B alpha was observed in the lungs of C3H/HeN at 4 h after treatment with LPS, while both the p-I kappa B alpha and I kappa B alpha contents showed a tendency to go down at 8 h after treatment and dramatically decreased at 24 h. But there were no changes in the of p-I kappa B alpha and I kappa B alpha content the lungs of C3H/HeJ under the same conditions. CONCLUSION: K. pneumoniea endotoxin could induce the expression of beta-defensin-4 mRNA in the lung of C3H/HeN, and TLR4-mediated NF-kappa B activation signaling pathway may be responsible for this event.

Amino Acid Sequence↗

PubMed2001

[Spectrophotometric determination of trace copper in electroplating wastewater].

In the presence of CPB and OP, the chromogenic reaction of DBH-PF with Cu(II) was studied. The results show that the purple micelle reaction has high sensitivity in the medium of (CH2)6N4-HCl(pH = 5.4) buffered solution, the molar absorptivity is 1.8 x 10(5) L.mol-1.cm-1, and Beer's law is obeyed for Cu(II) in the range of 0.0-9.0 micrograms.25 mL-1 at 540 nm. Combined with solvent extraction separation, the method has been applied to the determination of trace Cu in electroplating wastewater with satisfactory results.

Copper↗

PubMed2000

Isolation and characterization of disease resistance gene homologues from rice cultivar IR64.

We initiated a search for disease resistance (R) gene homologues in rice cultivar IR64, one of the most agronomically important rice varieties in the world, with the assumption that some of these homologues would correspond to previously identified disease resistance loci. A family of rice R gene homologues was identified using the Arabidopsis NBS-LRR disease resistance gene RPS2 as a hybridization probe. Because member genes of this rice R gene family exhibit features characteristic of the NBS-LRR class of resistance genes, the family was given the name NRH (for NBS-LRR resistance gene homologues). Three members of the NRH family, NRH1, NRH2, and NRH3, were cloned and studied in detail. In IR64, NRH1 and NRH2 appear to encode full-length polypeptides, whereas NRH3 is prematurely truncated with a stop codon generated by a frameshift. NRH1 maps on chromosome 5, and NRH2 and NRH3 are less than 48kb apart on chromosome 11. Although NRH1, NRH2, and NRH3 map to regions of the rice genome where disease resistance loci to Xanthomonas oryzae pv. oryzae (Xoo) have been identified, susceptible rice varieties transformed with either NRH1 or NRH2 failed to exhibit increased resistance to a set of well-characterized Xoo strains.

Amino Acid Sequence↗

PubMed2000

Mechanism of inhibition of the class A beta -lactamases PC1 and TEM-1 by tazobactam. Observation of reaction products by electrospray ionization mass spectrometry.

The reactions of class A beta-lactamases PC1 and TEM-1 with tazobactam (TZB), a potent penicillanic sulfone inhibitor for class A beta-lactamases, were studied using electrospray ionization mass spectrometry (ESI/MS). Following inactivation of the beta-lactamases by TZB, new abundant high mass components were observed including three with molecular masses of 52, 70, and 88 Da greater than PC1 and TEM-1, respectively, and a component with a molecular mass of 300 Da greater than PC1. In addition, three TZB reaction products with molecular masses of 248, 264, and 280 Da were observed. High performance liquid chromatography (HPLC)/ESI/MS analysis of the TZB-PC1 adduct digested with Glu-C revealed three new components with masses 52, 70, and 88 Da greater than that of the peptide composed of amino acid residues 58-82 and one new component with a mass 70 Da greater than that of the peptide composed of amino acid residues 125-141. HPLC/ESI/MS/MS analysis of the two digested peptides whose masses increased by 70 Da indicated that Ser-70 and Ser-130 were the most likely TZB-modified amino acid residues. Based on these data, a mechanism for the inactivation of the class A beta-lactamases by TZB is proposed. In this scheme, initial acylation of Ser-70 by TZB and opening of the lactam ring are followed by one of several different events: (1) the rapid decomposition of TZB with loss of the enamine moiety to form the propiolylated enzyme, (2) an intramolecular nucleophilic displacement of the imine or enamine moiety by Ser-130 to form a cross-linked vinyl ether, and (3) hydrolysis of the imine or enamines to form a Ser-70-linked aldehyde.

Chromatography, High Pressure Liquid↗

PubMed2000

Cloning of factors related to HIV-inducible LBP proteins that regulate steroidogenic factor-1-independent human placental transcription of the cholesterol side-chain cleavage enzyme, P450scc.

The cholesterol side-chain cleavage enzyme, cytochrome P450scc, initiates the biosynthesis of all steroid hormones. Adrenal and gonadal strategies for P450scc gene transcription are essentially identical and depend on the orphan nuclear receptor steroidogenic factor-1, but the placental strategy for transcription of P450scc employs cis-acting elements different from those used in the adrenal strategy and is independent of steroidogenic factor-1. Because placental expression of P450scc is required for human pregnancy, we sought factors that bind to the -155/-131 region of the human P450scc promoter, which participates in its placental but not adrenal or gonadal transcription. A yeast one-hybrid screen of 2.4 x 10(6) cDNA clones from human placental JEG-3 cells yielded two unique clones; one is the previously described transcription factor LBP-1b, which is induced by HIV, type I infection of lymphocytes, and the other is a new factor, termed LBP-9, that shares 83% amino acid sequence identity with LBP-1b. When expressed in transfected yeast, both factors bound specifically to the -155/-131 DNA; antisera to LBP proteins supershifted the LBP-9.DNA complex and inhibited formation of the LBP-1b.DNA complex. Reverse transcriptase-polymerase chain reaction detected LBP-1b in human placental JEG-3, adrenal NCI-H295A, liver HepG2, cervical HeLa, and monkey kidney COS-1 cells, but LBP-9 was detected only in JEG-3 cells. When the -155/-131 fragment was linked to a minimal promoter, co-expression of LBP-1b increased transcription 21-fold in a dose-dependent fashion, but addition of LBP-9 suppressed the stimulatory effect of LBP-1b. The roles of LBP transcription factors in normal human physiology have been unclear. Their modulation of placental but not adrenal P450scc transcription underscores the distinctiveness of placental strategies for steroidogenic enzyme gene transcription.

Amino Acid Sequence↗

PubMed2000

Gene therapy that inhibits nuclear translocation of nuclear factor kappaB results in tumor necrosis factor alpha-induced apoptosis of human synovial fibroblasts.

OBJECTIVE: Tumor necrosis factor alpha (TNFalpha) increases the survival and proliferation of human rheumatoid arthritis (RA) cell lines. These experiments were designed to determine if inhibition of nuclear factor kappaB (NF-kappaB) nuclear translocation leads to increased apoptosis of TNFalpha-treated human RA cell lines. METHODS: We constructed an inhibitor of nuclear factor kappaB(IkappaB) dominant-negative adenovirus (AdCMVIkappaB-DN) and an X-linked inhibitor of apoptosis (XIAP) antisense adenovirus (AdCMVXIAP-AS). Primary RA synovial fibroblast (RASF) cell lines were transfected in vitro, and SV40-transformed RA synovial cell lines in SCID mice were transfected in vivo. Cells were treated with TNFalpha and analyzed for apoptosis. RESULTS: There was no apoptosis of primary RASF transfected in vitro with AdCMVIkappaB-DN alone. In contrast, there was apoptosis of >85% of cells treated with AdCMVIkappaB-DN plus TNFalpha. Primary RASF in SCID mice also exhibited high levels of apoptosis after in vivo transfection with AdCMVIkappaB-DN followed by treatment with TNFalpha. There was no apoptosis after treatment with AdCMVIkappaB-DN in the absence of TNFalpha. XIAP is an inhibitor of apoptosis which was up-regulated by TNFalpha, and this up-regulation was inhibited by AdCMVIkappaB-DN plus TNFalpha. Transfection of an AdCMVXIAP-AS gene therapy resulted in increased TNFa-induced apoptosis. CONCLUSION: AdCMVIkappaB-DN gene therapy greatly enhances apoptosis due to inhibition of an NF-kappaB-mediated antiapoptosis signaling pathway, and XIAP is a TNFalpha-inducible specific inhibitor of apoptosis in RA synovial cell lines. This and other modulators of TNF receptor or the Fas apoptosis pathway may be therapeutically beneficial in facilitating apoptosis of synovial tissue in patients with RA.

Animals↗

PubMed2000

Pharmacological characterization of vasomotor activity of human musculocutaneous perforator artery and vein.

Vasospasm is one of the main causes of skin ischemic necrosis in cutaneous and musculocutaneous flap surgery, but the pathogenic mechanism is unclear. We planned to test the hypothesis derived from clinical impression that veins are more susceptible to vasospasm than arteries in flap surgery and, once established, that venous vasospasm is difficult to resolve and more detrimental than arterial vasospasm. To this end, we investigated the differences in sensitivity to vasoconstrictors and vasodilators between the human musculocutaneous perforator (MCP) artery and vein by measuring the isometric tension of arterial and venous rings suspended in organ chambers. Vascular contraction was expressed as a percentage of the tension induced by 50 mM KCl. Relaxation was expressed as a percentage of contraction induced by a submaximal concentration (3 x 10(-9) M) of endothelin-1 (ET-1). We observed that the vasoconstrictor potency of norepinephrine was significantly higher in the MCP vein than in the MCP artery. The vasoconstrictor potency of ET-1 and the thromboxane A(2) mimetic U-46619 were similar in the MCP vein and artery, but the maximal contraction induced by ET-1 and U-46619 was significantly higher in the MCP vein than in the MCP artery. On the other hand, the MCP vein was less sensitive than the MCP artery to the relaxation effect of nitroglycerin, nifedipine, and lidocaine. These differences between the human MCP artery and vein in response to vasoactive agents lend support to the clinical impression in flap surgery that veins appear to be more susceptible to vasospasm than arteries and venous vasospasm seems to be more difficult to resolve than arterial vasospasm in cutaneous and musculocutaneous flap surgery.

Arteries↗

PubMed2000

Quantitative trait loci for root-penetration ability and root thickness in rice: comparison of genetic backgrounds.

Drought is the major abiotic stress limiting rice (Oryza sativa) production and yield stability in rainfed lowland and upland ecosystems. Root systems play an important role in drought resistance. Incorporation of root selection criteria in drought resistance improvement is difficult due to lack of reliable and efficient screening techniques. Using a wax-petrolatum layer system simulated to compacted soil layers, root traits were evaluated in a doubled haploid (DH) population derived from the cross between 'IR64' and 'Azucena'. Twelve putative QTLs (quantitative trait loci) were detected by interval mapping comprising four QTLs for root-penetration ability, four QTLs for root thickness, two QTLs for penetrated root number, and two QTLs for total root number. These QTLs individually explained 8.4% to 16.4% of the phenotypic variation. No QTL was detected for maximum penetrated root length by interval mapping. One QTL located between RG104 and RG348 was found to influence both root-penetration ability and root thickness. QTLs for root-penetration ability and root thickness were compared across two populations, 'IR64'-'Azucena' and 'CO39'-'Moroberekan', and different testing conditions. The identified consistent QTLs could be used for marker-assisted selection for deep and thick roots with high root-penetration ability in rice.

Oryza↗

PubMed2000

[An experimental study of human tracheal epithelial transfer of human neutrophil defensin gene].

This study was amied at the possibility of production of the human neutrophil defensin in human tracheal epithelial cells when transfected with it's gene by using liposome. The plasmid vector carrying the human neutrophil defensin HNP1 (human Neutrophil Peptide-1) gene was used to transfect the human tracheal epithelial cells cultured in serum-free complement medium by lipofection reagent. Antiserum of HNP1 was prepared and used for the immunocytochemistry staining to detect expression of HNP1 gene in human tracheal epithelial cells. The result showed that recombinant eukaryote expression plasmid-pBabe-Neo-HNP1 could be transducted into human tracheal epithelial cells by liposomal transfection method. Transducted cells showed strong positive staining for HNP by immunohistochemistry assay. It was suggested that HNP1 gene could be productively transduced into human tracheal epithelial cells and expressed by using liposome. Antibiotic peptide gene might be used for gene therapy of respiratory infections.

Cells, Cultured↗

PubMed2000

[The neutralizing effect of histidine-rich-polypeptides on LPS from suspected periodontal pathogens].

OBJECTIVE: Human parotid histidine-rich-polypeptides (HRPs) are a family of lowmolecular-weight, cationic polypeptides. HRP-1, HRP-3 and HRP-5 comprise 85%-90% of the total HRPs and are called major HRPs. There were many researches indicating the antimicrobial activities of HRPs. Recently, HRPs were reported to have an inhibitory action on the lipopolysaccharide (LPS) of E. coli, with HRP-5 being the most effective one among the three major HRPs. Since the LPS of oral gram-negative bacteria is thought to be one of the important etiological factors during the development of periodontal diseases, our experiment was aimed to investigate the neutralizing effect of human parotid HRP-5 on the LPS of anaerobic suspected periodontal pathogens, which have different chemical structures and biological activities compared with LPS of aerobic E. coli. METHODS: By using of preparative acid urea polyacrylamide gel electrophoresis (AU-PAGE), HRP-5 was purified from parotid saliva collected from healthy adults. Two stains of suspected periodontal pathogens, Porphyromonas gingivalis (P.g, 47-A) and Fusobacterium nuceatum (F. n, separated from subgingival plaque of a patient with adult periodontitis) were mass cultured. After harvested in the midlogarithmic phase, they were washed and lyophilized. The LPS of dried bacteria were extracted by the modified Westphal hot phenol-water procedures and purified by the enzyme digestion plus ultracentrifugation. Limulus test was applied to test the neutralizing effect of HRP-5 on the LPS-induced gelation of Limulus amoebocyte lysate. In brief, the standard LPS of E. coli, or extracted LPS of P. g or F. n, was preincubated with HRP-5 in a tube separately in room temperature for 10 minutes. Then the reagent of Limulus amoebocyte lysate was added in all the tubes, continued the incubation in 37 degrees C for one hour. After that, the gelation level of every tube was observed. RESULTS: LPS extracted from P. g and F. n both showed good purity and strong activities to induce gelation of Limulus amoebocyte lysate. The gelation induced by LPS (1 ng/ml) of these two anaerobic suspected periodontal pathogens were weakly inhibited by HRP-5 (10 micrograms/ml), similar to that observed with standard LPS of E. coli. To get a complete neutralizing effect on LPS, it may be important to increase the concentration of HRP-5. CONCLUSION: It appeared that HRPs could neutralize the endotoxic properties of LPS of suspected periodontal pathogens, therefore may contribute to periodontal health. The present investigation further confirmed that HRPs are important components of the host non-immune defense system.

Escherichia coli↗