Biomedical subjects

L Xie

Publications and source records attributed to L Xie.

At least 19 recordsLinked to original sources

PubMed2006

Impact assessment of cadmium contamination on rice (Oryza sativa L.) seedlings at molecular and population levels using multiple biomarkers.

Assessment of environmental contamination on ecology (plant) at molecular and population levels is important in risk quantification and remediation study. Random amplified polymorphic DNA (RAPD) assay and related other fingerprinting techniques have been employed to detect the genotoxin-induced DNA damage and mutations. This research compared the effects occurring at molecular and population levels in rice seedlings exposed to cadmium (Cd) concentrations of 15-60 mg l(-1) for 8 days with quartz sand culture. Inhibition of root growth and increase of total soluble protein content in root tips of rice seedlings were observed with the increase of Cd concentration. For the RAPD analyses, 12 RAPD primers of 50-70% GC content were found to produce unique polymorphic band patterns and subsequently were used to produce a total of 180 bands of 179-3056 bp in molecular size in the control root tips of rice seedlings. Results produced by these RAPD primers indicate that changes in RAPD profiles of root tips after Cd treatment include modifications in band intensity and gain or loss of bands by comparison with control. The effect of changes was dose-dependent. Genomic template stability compares favourably with the traditional indices such as root growth and soluble protein content. The DNA polymorphisms detected by RAPD analysis can be applied as a suitable biomarker assay for the detection of genotoxic effects of Cd contamination on plants.

Biomarkers↗

PubMed2006

A distributed model of carbohydrate transport and metabolism in the liver during rest and high-intensity exercise.

A model of reaction and transport in the liver was developed that describes the metabolite concentration and reaction flux dynamics separately within the tissue and blood domains. The blood domain contains equations for convection, axial dispersion, and transport to the surrounding tissue; and the tissue domain consists of reactions representing key carbohydrate metabolic pathways. The model includes the metabolic heterogeneity of the liver by incorporating spatial variation of key enzymatic maximal activities. Simulation results of the overnight fasted, resting state agree closely with experimental values of overall glucose uptake and lactate output by the liver. The incorporation of zonation of glycolytic and gluconeogenic enzyme activities causes the expected increase in glycolysis and decrease in gluconeogenesis along the sinusoid length from periportal to perivenous regions, while fluxes are nearly constant along the sinusoid length in the absence of enzyme zonation. These results confirm that transport limitations are not sufficient to account for the observed tissue heterogeneity of metabolic fluxes. Model results indicate that changes in arterial substrate concentrations and hepatic blood flow rate, which occur in the high-intensity exercise state, are not sufficient to shift the liver metabolism enough to account for the 5-fold increase in hepatic glucose production measured during exercise. Changes in maximal activities, whether caused by exercise-induced changes in insulin, glucagon, or other hormones are shown to be needed to achieve the expected glucose output. This model provides a framework for evaluating the relative importance to hepatic function of various phenomenological changes that occur during exercise. The model can also be used to assess the potential effect of metabolic heterogeneity on metabolism.

Carbohydrate Metabolism↗

PubMed2006

Bicyclol for chronic hepatitis B.

BACKGROUND: Bicyclol is a novel synthetic 'anti-hepatitis' drug, used in China for chronic hepatitis B. Until now, systematic reviews of bicyclol therapy have not been performed. OBJECTIVES: To study the benefits and harms of bicyclol for patients with chronic hepatitis B. SEARCH STRATEGY: We searched The Cochrane Hepato-Biliary Group Controlled Trials Register (July 2005), The Cochrane Central Register of Controlled Trials in The Cochrane Library (Issue 2, 2005), MEDLINE (1950 to July 2005), EMBASE (1980 to July 2005), Science Citation Index Expanded (1945 to July 2005), The Chinese Biomedical Database (1994 to August 2005), VIP Chinese Science and Technique Journals Database (1994 to August 2005), and China National Infrastructure (CNKI)(1994 to August 2005). We also contacted manufacturers and researchers in the field. SELECTION CRITERIA: Randomised clinical trials with bicyclol versus no intervention, placebo, or other interventions were included, irrespective of blinding, publication status, or language. DATA COLLECTION AND ANALYSIS: The primary outcome measures were mortality (total and liver-related) and liver-related morbidity (eg, cirrhosis and carcinoma). Secondary outcome measures were viral response and liver histology. MAIN RESULTS: The search identified one randomised clinical trial comparing bicyclol with bifendate (biphenyldicarboxylate) for patients with hepatitis B. The follow-up was three months. There was no evidence that bicyclol was superior to bifendate for loss of HBeAg (RR 1.38, 95% CI 0.95 to 2.00), seroconversion of HBeAg to HBeAb (RR 1.44, 95% CI 0.90 to 2.29), loss of HBV DNA (RR 1.19, 95%CI 0.93 to 1.53), or number of patients with normalised alanine aminotransferase and aspartate aminotransferase activity (RR 0.88, 95% CI 0.70 to 1.11 and RR 0.97, 95% CI 0.79 to 1.20, respectively). AUTHORS' CONCLUSIONS: Only one randomised clinical trial has examined the potential benefit of bicyclol for patients with chronic hepatitis B. This small, short-term trial found no evidence to support or refute its use. Large, randomised double-blind clinical trials with long-term follow-up are needed to examine the possible benefits and harms associated with bicyclol. Bicyclol can only be recommended for use in randomised trials.

Biphenyl Compounds↗

PubMed2006

Organogenesis of pancreatic anlagen allografted in rats.

AIMS: To study the possibility of revascularization, growth, and differentiation of embryonic pancreatic anlagen transplanted to adult hosts. While transplantations of pancreas and islets are the main methods to cure diabetes mellitus, the donor source is in shortage. So it's necessary to find a new source for transplantation. METHODS: The pancreas from embryonic day 14.5 (E14.5) and 15.5 (E15.5) Lewis rat embryos were implanted into either intraperitoneal or subrenal capsular site of healthy Lewis rats. at 3 weeks or 6 weeks after implantation, the pancreatic anlagen in the host rats were resected for size measurements, as well as histopathologic and immunohistochemical examinations. RESULTS: Three weeks after implantation into the renal-capsular site, the size of both E14.5 and E15.5 pancreatic anlagen had enlarged 10- to 15-fold with differentiation of acinar components upon histological examination. Moreover, increasing numbers of beta cells and islets stained positive for insulin, and newly generated vessels were observed around the tissues. Continued proliferation of the endocrine islets in E14.5 pancreatic anlagen grafts was observed after another 3 weeks, whereas further proliferation in the E15.5 pancreatic anlagen graft was not seen. Additionally fibrosis appeared in the exocrine component of both E14.5 and E15.5 pancreatic anlagen at this time point. When implanted into intraperitoneal site, enlarged E15.5 pancreatic anlagen with proliferatels beta cells were also observed after 3 weeks. However, both the size of the pancreatic anlagen and the proliferation of the beta cells were much less than that in the subrenal capsular site. CONCLUSIONS: The allografted E14.5 and E15.5 pancreatic anlagen revascularised and grew into tissues that were structurally similar to normal mature rats pancreatic tissue. Adequate embryonic age for the transplantation of pancreatic anlagen is 14.5 and 15.5 days old. Subrenal capsula is a more suitable site than the peritoneal cavity for implantation of pancreatic anlagen.

Animals↗

PubMed2006

Prolonged cardiac allograft survival in presensitized rats after a high activity Yunnan-cobra venom factor therapy.

UNLABELLED: Complement-dependent antibody-mediated acute humoral rejection is the major obstacle of clinical transplantation across ABO incompatibility and human leukocyte antigen presensitization. We previously demonstrated that Yunnan-cobra venom factor (Y-CVF) could almost completely abrogate complement activity and successfully prevent hyperacute rejection in some xenotransplant models without any obvious toxicity. In this study we investigated whether depletion of complement by Y-CVF prevented acute humoral allograft rejection in presensitized rats thereby prolonging graft survival. METHODS: Presensitization was achieved in Lewis rats by sequential grafting of three full-thickness skin pieces from Brown Norway rats. Serum cytotoxic alloantibody titers were determined by a modified in vitro complement-dependent microcytotoxicity assay. After presensitization, each Lewis rat received a heterotopic Brown Norway cardiac allograft. Fifteen recipients were divided into two groups: (1) no treatment control (n = 7); (2) Y-CVF therapy group (86 u/kg, IV, day -1) (n = 8). After cessation of the heart beat, allograft rejection was confirmed by pathologic as well as IgG and C3 immunohistochemical examinations. RESULTS: The mean graft survival time was significantly prolonged to 99.50 +/- 38.72 hours among rats that received Y-CVF vs 12.71 +/- 13.94 hours in nontreated controls (P < .001). Upon pathological and immunohistochemical examination, acute humoral rejection was mainly exhibited in the control group, whereas acute cellular rejection was mainly displayed in the Y-CVF therapy group. CONCLUSIONS: Our study demonstrated that complement depletion by Y-CVF significantly inhibited acute humoral allograft rejection in presensitized rats. As a therapeutic immunointervention tool for complement, Y-CVF has shown potential efficacy across ABO incompatible and positive cross-match barriers.

Animals↗

PubMed2006

Sertoli cells induce xenolymphocyte apoptosis in vitro.

BACKGROUND: Testicular Sertoli cells can protect pancreatic islet grafts from allo- and autoimmune destruction; however, the mechanisms underlying immune privilege of the testicle are not well understood, especially in xenotransplantation. The purpose of this study was to investigate whether rat Sertoli cells could induce mouse lymphocyte apoptosis in vitro. METHODS: Testis was isolated from 2- to 4-week-old Sprague Dawley (SD) rats. Sertoli cells were successfully prepared by digestion with collagenase type V, trypsin, and DNase I, and then identified by electron microscope. Viability and apoptosis of cultured cells were measured by flow cytometry. We examined the apoptosis rates of Balb/c mouse lymphocytes, which were cocultured with SD rat Sertoli cells by FACS. The expression of Fas ligand (Fasl), transforming growth factor (TGF)-beta1 and clusterin on Sertoli cells were detected by immunocytochemistry. RESULTS: In the cocultured system, Sertoli cells accounted for more than 93%. With our isolation method, the viability of Sertoli cells was more than 95% and the apoptosis rate was 10.87% +/- 3.87%. The lymphocyte apoptosis ratio was 15.52 +/- 0.17 (P < .01, compared with the control groups). SABC immunochemistry staining showed that the sertoli cells could express FasL, TGF-beta, and clusterin. CONCLUSIONS: In our coculture in vitro, rat Sertoli cells expressed FasL and TGF-beta1 as well as induced the apoptosis of mouse lymphocytes. These results indicated that the expression of FasL and TGF-beta1 on Sertoli cells might relate to immune privilege in xenotransplantation.

Animals↗

PubMed2006

A study of soluble HLA-G1 protecting porcine endothelial cells against human natural killer cell-mediated cytotoxicity.

UNLABELLED: Human natural killer (NK) cells, which can mediate direct lysis of porcine endothelial cells, play an important role in xenograft rejection. HLA-G, which is a critical molecule in maintaining maternal immune tolerance of semi-allogenic fetus, is able to protect susceptible target cells from lysis induced by NK cells. In this study, we investigated whether soluble HLA-G1 (sHLA-G1) protected porcine xenogeneic cells against human NK cell-mediated lysis. METHODS: The human sHLA-G1 genomic DNA (pcDNA3-sHLA-G1) was transfected into a B lymphoblastoid cell line 721.221 (LCL721.221) by nucleofector. The sHLA-G1 expression of the transfected LCL721.221 cells was identified by RT-PCR and Dot-ELISA. The sHLA-G1 protein was purified by affinity chromatography on anti-HLA-ImAb W6/32 coupled to cyanogen-bromide-activated Sepharose 4B from culture supernates of transfectants. Various concentrations of sHLA-G(1) protein (0, 2, 4, 6, or 8 microg/mL) were added to a NK cell-mediated xenogenic cell lysis system with either NK92 cells or fresh human peripheral blood mononuclear cells (PBMCs) cocultured with the porcine endothelial cells line. A LDH release assay was used to evaluate NK cell-mediated cytotoxicity. RESULTS: sHLA-G1 provided significant protection of porcine endothelial cells against human NK-mediated cytotoxicity in a dose-dependent manner. The rates of NK92 cell-mediated cytotoxicity were reduced to 83.4 +/- 5.7% (2 microg/mL), 56.6 +/- 9.3% (4 microg/mL), 39.3 +/- 10.2% (6 microg/mL), and 31.2 +/- 4.9% (8 microg/mL) versus 96.9 +/- 3.0% in the control group (P < .01). Similarly, adding 6 microg/mL sHLA-G1 reduced the mean rate of PBMC-mediated cytotoxicity (n = 4) to 5.8 +/- 1.6% from 23.9 +/- 1.3% in the control group (P < .01). CONCLUSIONS: These results indicated that sHLA-G1 protected xenogeneic porcine endothelial cells against attack by human NK cells, thus providing a new approach to overcome NK-mediated immunity to xenografts.

Animals↗

PubMed2006

Long-term outcome of combined penetrating keratoplasty and vitreoretinal surgery using temporary keratoprosthesis.

PURPOSE: To evaluate the long-term outcome of combined penetrating keratoplasty (PKP) and vitreoretinal surgery using temporary keratoprosthesis (TKP). METHODS: A retrospective study of 107 eyes with coexisting corneal and vitreoretinal diseases that underwent combined PKP and vitreoretinal surgery using TKP. Corneal graft clarity, intraocular pressure, and anatomical reattachment of retina were followed. RESULTS: The mean follow-up time was 25 months, the longest being 8 years. Successful surgical outcome was defined as maintenance of clear graft, anatomic reattachment of retina, and controlled intraocular pressure. A total of 78 eyes (72.9%) fulfilled these criteria. Surgical intervention within 1 month of ocular injury was associated with higher success rate (81.9%) than intervention at 1 month or more after injury (54.3%). Success rate for reattachment of retina was 95.8% when preoperative proliferative vitreoretinal retinopathy (PVR) was absent compared with 83.1% when preoperative PVR was present. Unsatisfactory postoperative visual acuity was due to graft failure, recurrent PVR, or secondary glaucoma. CONCLUSION: Combined PKP and vitreoretinal surgery is best performed within 1 month of ocular injury for best surgical outcome. Careful selection of cases plays an important role in reducing the risks of complications. TKP is a useful adjunct in the surgery.

Adolescent↗

PubMed2006

Comparison of bovine in vivo bioavailability of two sulfamethazine oral boluses exhibiting different in vitro dissolution profiles.

The bolus (or oblet) is a dosage form that can be used for the oral administration of pharmaceutical compounds to ruminating species. Unlike traditional tablets, oral boluses may contain quantities of drug on the order of grams rather than milligrams. Due to its size, it is only recently that USP-like in vitro dissolution methods have been developed for this dosage form. However, whether or not these dissolution tests can predict product in vivo performance has yet to be determined. The importance of this issue is apparent when the U.S. Food and Drug Administration Center for Veterinary Medicine is faced with the decision of whether to require additional in vivo bioequivalence study data to support the approval of changes in product chemistry or manufacturing method. The current study was undertaken to determine whether an in vivo/in vitro correlation can be established for bovine sulfamethazine oral boluses and to acquire insight into the magnitude of changes in in vitro product performance that can occur before corresponding changes are seen in in vivo blood level profiles. Based upon the results of this investigation, it is concluded that marked changes in in vitro sulfamethazine bolus performance can be tolerated before resulting in altered in vivo blood level profiles. However, the data also suggest that rumenal absorption may occur for some compounds. Therefore the degree to which variation in product in vitro dissolution profiles can be tolerated may be compound specific.

Administration, Oral↗

PubMed2006

Naloxone and epinephrine are equally effective for cardiopulmonary resuscitation in a rat asphyxia model.

BACKGROUND: It is not known whether naloxone is as efficacious as epinephrine during cardiopulmonary resuscitation (CPR). The aim of the study was to compare the effects of naloxone and epinephrine on the outcomes of CPR following asphyxial cardiac arrest in rats. METHODS: Cardiac arrest was induced with asphyxia by clamping the tracheal tubes. Twenty-four Sprague-Dawley rats were randomized prospectively into a saline group (treated with normal saline, 1 ml intravenously, n = 8), an epinephrine group (treated with epinephrine, 0.04 mg/kg intravenously, n = 8) or a naloxone group (treated with naloxone, 1 mg/kg intravenously, n = 8) in a blind fashion during resuscitation after asphyxial cardiac arrest. After 5 min of untreated cardiac arrest, conventional manual CPR was started and each drug was administered at the same time. RESULTS: The rates of restoration of spontaneous circulation (ROSC) were one of eight (12.5%), seven of eight (87.5%) and seven of eight (87.5%) in the saline, epinephrine and naloxone groups, respectively. The rates of ROSC in the epinephrine and naloxone groups were equal and significantly greater than that in the saline group (P = 0.01 and P = 0.01, respectively). CONCLUSION: The administration of naloxone or epinephrine alone may increase the resuscitation rate, and both drugs are equally effective for CPR in a rat asphyxia model. However, the mechanism by which naloxone produces its efficacy during CPR remains unclear and further experimentation will be necessary.

Animals↗

PubMed2005

Evaluation of wetland and tertiary wastewater treatments for estrogenicity using in vivo and in vitro assays.

The effects of wetland wastewater treatment on estrogenic activity and estrogenic activity of water after tertiary treatment were evaluated using in vivo (rainbow trout vitellogenin [VTG] expression) and in vitro (yeast estrogen screening) assays. Juvenile rainbow trout exposed to tertiary-treated wastewater from the Green Acres Treatment Plant in Orange County Water District had increased plasma VTG levels compared with control fish. When trout were exposed to wastewater-dominated water before it entered into Prado Wetland (Riverside County, CA), VTG concentrations were increased above those of controls and were not significantly different from fish exposed to water exiting the wetland. VTG E2-equivalent concentrations (EEQs) of the water samples from the Green Acres Plant were 16.92 +/- 16.49 ng/L. Activity of water entering Prado Wetland was 29.80 +/- 28.41 ng/L EEQ, and water exiting was 24.34 +/- 23.17 ng/L EEQ. In vitro assays estimated that estrogenic activity of water from the Green Acres Plant was <1 ng/L EEQ, whereas water entering and exiting Prado Wetland had yeast estrogen screening EEQs of 2.57 and <1 ng/L, respectively. Our results suggest that environmental estrogens that are not potent estrogen-receptor ligands exist in wastewaters from the Green Acres Plant as well as water entering and exiting Prado Wetland. Wetland treatment did not remove environmental estrogens in the water. Our results also suggest that in vitro assays may underestimate estrogenic activity of sampled water.

Animals↗

PubMed2005

A study of neonatal swimming (water therapy) applied in clinical obstetrics.

OBJECTIVE: To study the significance of some clinical parameters related to neonatal 'swimming' (water therapy) during hospitalization. METHODS: Normal newborns were randomly divided into two groups to observe their birth weight, weight before discharge,time of first defecation and meconium turning yellow. Group one was the swimming (study) group, comprising a total of 223 newborns including 127 babies delivered after spontaneous vaginal delivery and 96 babies after Cesarean section. Group two was the bathing (control) group, comprising 154 newborns including 109 babies delivered after spontaneous vaginal delivery and 45 babies after Cesarean section. RESULTS: There was no significant difference in birth weight between the two groups (p > 0.05). However, the mean weight before discharge of the babies in the study group was 3.29 + 0.35 and 3.51 + 0.40 kg, spontaneous vaginal delivery vs. Cesarean section, compared with 3.09 + 0.38 and 3.17 + 0.48 kg, respectively, in the control group (p < 0.01). The corresponding mean times of meconium turning yellow were 39.15 + 15.88 and 39.02 + 13.60 h in the study group compared with 48.01 + 19.42 and 55.67 + 25.05 h in the control group. This difference was significant (p < 0.01), as was the difference between the time of first defecation (p < 0.05). CONCLUSION: Neonatal swimming can accelerate babies' growth in the early stage.

Baths↗

PubMed2005

Survival patterns for the top four cancers in Canada: the effects of age, region and period.

This study examined the variations in survival rates (1989-1991) and the trends (1969-1991), by sex, age and province, for patients diagnosed with breast, colorectal, lung or prostate cancer in Canada and compared the Canadian rates with those of nine American SEER registries. Five-year age-standardized relative survival rates (ASRs) were calculated, and the trends were estimated from variance-weighted linear regression of the ASRs for five periods of diagnosis (1969-1973, 1974-1978, 1979-1983, 1984-1988 and 1989-1991). In 1989-1991, the ASR varied among provinces for each cancer except female colorectal cancer. The lowest survival rates were observed in the youngest patients (15-44) for breast and prostate cancers, and in the oldest patients (75-99) of both sexes for lung and colorectal cancers. Over the five periods, a major trend toward improved survival was observed for breast, prostate and colorectal cancers (P<0.008), whereas no changes were seen for lung cancer. The ASRs in the western region were higher than in the Atlantic region over time (P<0.02) for each cancer. From the third period onward, the ASRs for Canadian patients with lung cancer were similar to those for the US patients and lower than for Canadian patients with breast, prostate or colorectal cancer. The observed increases in ASR for breast and prostate cancer are likely due to the increased use of screenings and the improved treatment modalities.

Adolescent↗

PubMed2004

Survival of women with breast cancer in Ottawa, Canada: variation with age, stage, histology, grade and treatment.

This study examined the 5-year survival of 2192 breast cancer women diagnosed between 1994 and 1997 in Ottawa, Canada, by age, TNM stage, histology, grade and treatment, including assessment of the independent value of variables in defining prognosis. Our results showed that age, stage, treatment and grade significantly influenced outcome regardless of the confounding factors considered, with histology failing to achieve significant independent prognostic information. The survival rates were highest at ages 50-69 years for stage I and at ages 40-49 years for stages II-IV. The rates were lowest at ages or=70 years for stages III-IV. The differences in survival between grade 1 and grade 3 were 9% in stage I and 20% in stage II. The treatment leading to the best survival was surgery plus radiation for stages I-II and surgery combined with chemotherapy for stages III-IV. Lobular carcinoma had a better prognosis than ductal carcinoma; this can be explained by more grade 1 and less grade 3 cases in lobular carcinoma. The worse prognosis for young patients than other ages can be explained by their higher proportion of poorly differentiated cancers. Stage I patients aged 50-69 years having the best survival is likely due to the earlier diagnosis achieved through screening.

Adult↗

PubMed2004

The CLDN5 locus may be involved in the vulnerability to schizophrenia.

The present study was designed to detect three single nucleotide polymorphisms (SNPs) located on 22q11 that was thought as being of particularly importance for genetic research into schizophrenia. We recruited a total of 176 Chinese family trios of Han descent, consisting of mothers, fathers and affected offspring with schizophrenia for the genetic analysis. The transmission disequilibrium test (TDT) showed that of three SNPs, rs10314 in the 3'-untranslated region of the CLDN5 locus was associated with schizophrenia (chi(2) = 4.75, P = 0.029). The other two SNPs, rs1548359 present in the CDC45L locus centromeric of rs10314 and rs739371 in the 5'-flanking region of the CLDN5 locus, did not show such an association. The global chi-square (chi(2)) test showed that the 3-SNP haplotype system was not associated with schizophrenia although the 1-df test for individual haplotypes showed that the rs1548359(C)-rs10314(G)-rs739371(C) haplotype was excessively non-transmitted (chi(2) = 5.32, P = 0.02). Because the claudin proteins are a major component for barrier-forming tight junctions that could play a crucial role in response to changing natural, physiological and pathological conditions, the CLDN5 association with schizophrenia may be an important clue leading to look into a meeting point of genetic and environmental factors.

Cell Cycle Proteins↗

PubMed2004

No association between the PTGS2/PLA2G4A locus and schizophrenia in a Chinese population.

The present study was undertaken to replicate an association between the PTGS2/PLA2G4A locus and schizophrenia among a Chinese population. We recruited 168 Chinese parent-offspring trios of Han descent, consisting of fathers, mothers and affected offspring with schizophrenia. Of 3 informative SNPs genotyped, no one showed allelic association with schizophrenia; the haplotype analysis also failed to capture a haplotypic association with the illness. Because the frequencies of alleles and genotypes of SNPs analyzed differ in the Chinese population as compared with a British population that initially showed the genetic association between the PTGS2/PLA2G4A locus and schizophrenia, the ethnic background may be a major reason for poor replication of the initial finding.

Asian People↗

PubMed2003

Optimization of recombinant aminolevulinate synthase production in Escherichia coli using factorial design.

The production of recombinant Rhodobacter sphaeroides aminolevulinate (ALA) synthase was optimized in two strains of Escherichia coli: the wild-type strain MG1655, and a ptsG mutant AFP111. The effects of initial succinate, glucose and isopropyl-beta-d-thiogalactopyranoside (IPTG) concentrations and the time of induction on enzyme activity were studied. One-way analysis was used to approximate the optimal ranges for these factors, followed by a full factorial design to quantify the effects of each factor and the interactions between the factors. Initial succinate, glucose, and IPTG concentration were observed to be the key factors affecting ALA synthase activity with the optimal levels determined to be above 6 g/l succinate, 0 g/l glucose, and 0.10 mM IPTG. ALA synthase activity was generally lower with AFP111 than with MG1655, and the effect of these three key factors was also lower with AFP111 than with MG1655. Based on the full factorial design results, a fermentation was completed that yielded 296 mU/mg protein with a final ALA concentration of 5.2 g/l (39 mM).

5-Aminolevulinate Synthetase↗

PubMed2003

Production of 5-aminolevulinic acid by an Escherichia coli aminolevulinate dehydratase mutant that overproduces Rhodobacter sphaeroides aminolevulinate synthase.

Two Escherichia coli strains, a wild-type strain and a hemB mutant, which contained the hemA gene from Rhodobacter sphaeroides produced aminolevulinate at 4 g l(-1). The hemB mutant did not increase the accumulation of aminolevulinate, an observation attributed to its 50% lower aminolevulinate synthase activity in than the wild-type. Growth was limited for both strains probably due to the accumulation of 0.5 g aminoacetone l(-1).

5-Aminolevulinate Synthetase↗