Engineering Saccharomyces cerevisiae for efficient production of glucaric acid / 生物工程学报

As an important dicarboxylic acids existing in nature, glucaric acid has been widely used in medical, health, and polymer materials industry, therefore it is considered as one of the "top value-added chemicals from biomass". In this study, using Saccharomyces cerevisiae as a chassis microorganism, the effects of overexpression of myo-inositol transporter Itr1, fusional expression of inositol oxygenase MIOX4 and uronate dehydrogenase Udh, and down-expression of glucose-6-phosphate dehydrogenase gene ZWF1 on the glucaric acid production were investigated. The results showed that the yield of glucaric acid was increased by 26% compared with the original strain Bga-3 under shake flask fermentation after overexpressing myo-inositol transporter Itr1. The yield of glucaric acid was increased by 40% compared with Bga-3 strain by expressing the MIOX4-Udh fusion protein. On these basis, the production of glucaric acid reached 5.5 g/L, which was 60% higher than that of Bga-3 strain. In a 5 L fermenter, the highest yield of glucaric acid was 10.85 g/L, which was increased 80% compared with that of Bga-3 strain. The application of the above metabolic engineering strategy improved the pathway efficiency and the yield of glucaric acid, which may serve as a reference for engineering S. cerevisiae to produce other chemicals.

Construction of a glucaric acid biosensor for screening myo-inositol oxygenase variants / 生物工程学报

Glucaric acid (GA), a top value-added chemical from biomass, has been widely used for prevention and control of diseases and the production of polymer materials. In GA biosynthesis pathway, the conversion of inositol to glucuronic acid that catalyzed by myo-inositol oxygenase is the limiting step. It is necessary to improve MIOX activity. In the present study, we constructed a high-throughput screening system through combing the concentration of GA with the green fluorescent protein fluorescence intensity. By applying this screening system, three positive variants (K59V/R60A, R171S and D276A) screened from the mutant library. In comparison, the recombinant strain Escherichia coli BL21(DE3)/MU-R171S accumulated more GA, 136.5% of that of the parent strain.

Efficacy of intravenous iron sucrose infusion in children with iron deficiency anemia: experience at children hospital and ICH Multan

To describe the efficacy of intravenous iron sucrose in children with iron deficiency anemia who did not respond to /tolerate oral iron therapy. Quasi experimental interventional study. A prospective study was performed in 100 children, aged between 08 months and 15 years, 66% male and 33% female, diagnosed as iron deficiency anemia with predefined criteriaexcluding other causes. The children who did not respond to /tolerate oral iron therapy were treated with injectable ironin a day care hematology center. Dose of iron sucrose was calculated by a formula. Total dose was divided in three equal aliquots, each one was diluted in 0.9% normal saline and infused over a period of 120 minutes onthree consecutive days. The efficacy of iron sucrose was analyzed by comparing baseline mean hemoglobin at initiation of therapy and mean hemoglobin level two weeks after iron infusion. Mean age was 4.18 +/- 3.68 years.At start of treatment,baseline meanHb was 6.09 +/- 1.37 g/dl, mean MCV 51.5 +/- 9.03 fl and mean ferritin 7.76 +/- 7.6232 ng/ml. At day 14, mean Hb was 9.21 +/- 1.134 g/dl [P < 0.05], mean MCV 66.5 +/- 7.19 fl and mean ferritin 52.47 +/- 29.6828 ng/ml . Mean hemoglobin rise was 3.12 +/- 1.081 gm/dl [P < 0.05]. During infusion, only one patient had hypotensionwho was treated. Iron sucrose infusion is a safe and effective way of raising hemoglobin in iron deficiency anemia in pediatric age group in special set up with minimal side effects

Progress in glucaric acid / 生物工程学报

Glucaric acid (GA) is derived from glucose and commonly used in chemical industry. It is also considered as one of the "Top value-added chemicals from biomass" as carbohydrate monomers to produce various synthetic polymers and bioenergy. The demand for GA in food manufacture is increasing. GA has also attracted public attentions due to its therapeutic uses such as regulating hormones, increasing the immune function and reducing the risks of cancers. Currently GA is produced by chemical oxidation. Research on production of GA via microbial synthesis is still at preliminary stage. We reviewed the advances of glucaric acid applications, preparation and quantification methods. The prospects on production of GA by microbial fermentation were also discussed.

Intravenous Iron Sucrose for Three Children with Iron Deficiency Anemia Failing to Respond to Oral Iron Therapy / 임상소아혈액종양

Iron deficiency anemia is still a common pediatric hematologic disease in the world. First line treatment for iron deficiency anemia is oral iron supplementation. However, some children need parenteral iron therapy either because they cannot receive oral iron or because they do not respond to it. We used intravenous iron sucrose for three children with iron deficiency anemia who did not respond to oral iron. All were successfully treated without any significant adverse effects. Since studies about effects and adverse effects of intravenous iron sucrose on pediatric patients with iron deficiency anemia are limited, further studies are needed.

Intravenous Iron Sucrose for Three Children with Iron Deficiency Anemia Failing to Respond to Oral Iron Therapy / 임상소아혈액종양

Iron deficiency anemia is still a common pediatric hematologic disease in the world. First line treatment for iron deficiency anemia is oral iron supplementation. However, some children need parenteral iron therapy either because they cannot receive oral iron or because they do not respond to it. We used intravenous iron sucrose for three children with iron deficiency anemia who did not respond to oral iron. All were successfully treated without any significant adverse effects. Since studies about effects and adverse effects of intravenous iron sucrose on pediatric patients with iron deficiency anemia are limited, further studies are needed.

Comparison of Oral and Intravenous Iron for Treatment of Iron Deficiency Anaemia in Pregnancy.

Oral iron supplementation is standard obstetrical practice. But programmes of anaemia prophylaxis based on oral iron supplementation still remain a failure. Whether or not there is need for parenteral iron may be debated. Parenteral iron is indicated when oral iron supplementation is not possible or fails. Intravenous iron could be a good alternative in pregnant women to ensure compliance. Many workers have shown that parenteral iron therapy supplies enough iron for erythropoiesis. Therefore, In the present review, we assessed oral iron supplementation with intravenous (IV) iron sucrose in the management of iron deficiency anaemia in pregnancy. Intravenous iron sucrose appears a good alternative to oral iron in patients who are either intolerant to oral iron or are non compliant and may also reduce the incidence of blood transfusion and its associated risks.

Multi-frequency low-dose intravenous iron on oxidative stress in maintenance hemodialysis patients / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of different intravenous iron treatment regimens on anemia and oxidative stress in maintenance hemodialysis (MHD) patients.@*METHODS@#A total of 58 MHD patients were randomly divided into a multi-frequency low-dose intravenous iron group (iron sucrose 25 mg, twice a week for 8 weeks, n=19), a less-frequency regular-dose intravenous iron group (iron sucrose 100 mg, once every two weeks for 8 weeks, n=19), and a non-iron group (n=20). Another 20 healthy people served as a control group (n=20). The changes of hemoglobin (Hb), hematocrit (HCT), serum ferritin (SF) and transferrin saturation (TSAT), as well as the oxidative stress parameters of malon-dialdehyde (MDA), superoxide dismutase (SOD) and myeloperoxidase (MPO) were detected before and after the treatment.@*RESULTS@#After 8 weeks, compared with the non-iron group, the levels of Hb, HCT, SF and TSAT in the two iron groups were significantly elevated (P0.05). After the single dialysis, the two iron groups had higher level of serum MDA, MPO and lower level of serum SOD than that of the non-iron supplementation group (P0.05).@*CONCLUSION@#Multi-frequency low-dose intravenous iron can effectively improve anemia in MHD patients, whose acute oxidative stress is lower than that of less-frequency regular-dose intravenous iron, and is a relatively safe and effective intravenous iron treatment regimen.

Invertase, glucose oxidase and catalase for converting sucrose to fructose and gluconic acid through batch and membrane-continuous reactors / Invertase de glicose oxidase e catalase para a conversão de sacarose para ácido glucónico e frutose através de lote e de membrana-contínuas reactores

Conversion of sucrose into fructose and gluconic acid using invertase, glucose oxidase and catalase was studied by discontinuous (sequential or simultaneous addition of the enzymes) and continuous (simultaneous addition of the enzymes in a 100 kDa-ultrafiltration membrane reactor) processes. The following parameters were varied: concentration of enzymes, initial concentration of substrates (sucrose and glucose), pH, temperature and feeding rate (for continuous process). The highest yield of conversion (100 percent) was attained through the discontinuous (batch) process carried out at pH 4.5 and 37 ºC by the sequential addition of invertase (14.3 U), glucose oxidase (10,000 U) and catalase (59,000 U).

Neste trabalho estudou-se a conversão da sacarose em frutose e ácido glicônico, usando as enzimas invertase, glicose oxidase e catalase, através do emprego de processo descontínuo (com adição sequencial ou simultânea das enzimas) e contínuo (adição simultânea das enzimas em reator com membrana acoplado à membrana de ultrafiltração de 100 kDa). Os parâmetros variados foram: a concentração das enzimas, a concentração inicial dos substratos (sacarose e glicose), o pH, a temperatura e a vazão específica de alimentação (processo contínuo). Obteve-se rendimento de 100 por cento, quando a conversão foi conduzida por processo descontínuo em pH 4,5 e a 37 ºC com adição seqüencial das enzimas invertase (14,3 U), glicose oxidase (10.000 U) e catalase (59.000 U).

Iron deficiency anemia in childhood

Iron deficiency anemia (IDA) frequently occurs in infants and adolescents. IDA is the result of an interplay between increased host requirements, limited external supply, and increased blood loss. In outpatient clinics, we often see children with iron deficiency anemia. Most cases in children are caused by incomplete nutrient supplements and growth spurts. However, we can occasionally see patients with poor response despite iron supplementation. Failure of iron therapy occurs when a child does not receive the prescribed medication, when iron is given in a form that is poorly absorbed, or when there is a continuing unrecognized blood loss such as intestinal or pulmonary loss, or loss with menstrual periods. In addition, the therapeutic failure of iron medication may indicate that the original diagnosis of nutritional iron deficiency was incorrect. In this situation, we have to evaluate other etiologies of anemia. Recently, many cases relating H.pylori infection to iron deficiency anemia have been described in the literature and H.pylori infection has emerged as a cause of refractory iron deficiency anemia that is unresponsive to oral iron therapy. Also, iron deficiency anemia induced by athletics in adolescent females has been reported several times. In this article, the author reviews various etiologies of childhood iron deficiency anemia. The most important consideration in treatment of iron deficiency anemia is disclosure of the underlying cause and its recovery. Dietary habits should also be corrected. To supplement iron, 6 mg/kg of oral iron supplements (elemental iron) is recommended in ferrous salt form. If oral administration is not feasible, intravenous supplementation is recommended using forms such as iron dextran, iron gluconate, or iron sucrose.

The Effects of Preoperatively Administering Parenteral Iron Sucrose in Patients Who Are Undergoing Total Knee Arthroplasty

PURPOSE: The purpose of this study was to evaluate the efficacy of preoperative parenteral iron sucrose therapy in patients with postoperative anemia following total knee arthroplasty. MATERIALS AND METHODS: We retrospectively reviewed 30 patients who had postoperative anemia and who had undergone total knee arthroplasty from March 2008 to November 2008. Those patients agreed on treatment with preoperative parenteral iron sucrose therapy. The other 30 patients without iron sucrose administration were selected as a control group from January 2007 to February 2008. In this study, age, gender, the preoperative and postoperative values of hemoglobin, the operation time, the amount of bleeding, the amount of transfusion, the frequency of transfusion, postoperative infection and the length of the hospital stay were evaluated and statistically compared. Postoperative complications were also assessed. RESULTS: Ant complications due to the administration of iron sucrose were not observed. At 48 hours postoperatively, the mean hemoglobin of iron sucrose administration group was 9.4 g/dL and that of the control group was 8.7 g/dL (p=0.008). Homogenous transfusion was done for 17 cases in the iron sucrose administration group and for 22 cases in the non-iron sucrose administration group, but among these groups, the results were statistically insignificant. The amount of homogenous transfusion for each patient who underwent iron sucrose administration is 1.3 units and this was 1.7 units for each patient who underwent non-iron sucrose administration. No statistical differences were observed between the groups in this study. CONCLUSION: Parenteral iron sucrose administration in patients following total knee arthroplasty was not efficacious for preventing immediate postoperative anemia.

Intravenous Iron Supplementation in Korean Children on Chronic Dialysis

PURPOSE: Limited information is available on experiences of intravenous iron treatment in children. In this study, iron sucrose was administered intravenously to determine its effect, the factors predicting outcome, and safety in children on chronic dialysis. METHODS: Twenty-one children whose serum ferritin levels were less than 100 ng/mL or transferrin saturations (TSAT) were less than 20% were enrolled. In 12 children on peritoneal dialysis (PD), the drug was infused intravenously as 200 mg/m2 (< or =200 mg) at week 0, 2, 4, and 6. In 9 children on hemodialysis (HD), it was given intravenously as 8 weekly doses of 3 mg/kg (< or =100 mg) through week 0-7. RESULTS: After treatment, serum ferritin levels increased significantly in both groups, and TSAT rose significantly in PD group. However, hemoglobin level did not rise significantly in both groups. Children with baseline hemoglobin less than 10 g/dL or baseline TSAT less than 20% showed significantly higher rise of hemoglobin after intravenous iron treatment. To the contrary, those with higher baseline hemoglobin and TSAT levels displayed higher rise in serum ferritin after the treatment. Although no serious adverse event occurred, TSAT levels exceeding 50% were noted in 6 patients in PD group. CONCLUSION: This suggests that 3 mg/kg/week of intravenous iron sucrose can be used safely in children on chronic HD, but 200 mg/m2 every other week may incur excessive TSAT level in some patients on chronic PD.

Efficacy and Safety of Intraveous Iron Sucrose in the Perioperatively Anemic Patients of Gastrectomy

PURPOSE: This study was aimed to evaluate the efficacy and safety of of intravenous iron sucrose (Venoferrum(R)) for treating the perioperative anemic gastrectomy patients. MATERIALS AND METHODS: From September 2006 to February 2007 at Seoul National University Hospital, the gastrectomy patients who displayed perioperative anemia (7.0 g/dl or = 1 month after gastrectomy) patients without evidence of acute bleeding were included into Group 1. The immediate postoperative (< 1 month after gastrectomy) patients with stable vital signs were included into Group 2. The age, gender, diagnosis, Hb, hematocrit (Hct), mean corpuscular volume (MCV), serum ferritin (SF), total iron binding capacity (TIBC), serum iron and reticulocyte counts (RC) were evaluated before and after intravenous iron sucrose administration. The adverse effects of drugs were investigated. RESULTS: The number of patients of group 1 and group 2 was 79 and 46, respectively. In group 1, there was a statistically significant difference in the Hb, Hct, MCV, SF, RC and TIBC with each mean change of 1.3 g/dl, 4.1%, 3.1fl, 195 ng/ml, 0.2% and -86.4 ug/dl, respectively. In group 2, there was a statistically significant difference in the Hb, Hct, MCV, SF and RC with each mean change of 1.8 g/dl, 6.1%, 3.4fl, 260 ng/ml and 0.3%, respectively. Two patients (1.6%) suffered local thrombophlebitis as an adverse effect. CONCLUSION: Intravenous iron sucrose for the perioperative anemia of gastrectomy patients was efficacious in the short period without significant adverse effects.

Efficacy of Intravenous Iron Sucrose for Treating Anemia after Gastrectomy

PURPOSE: Many patients suffer with anemia after gastrectomy. Iron deficiency due to a decreased oral intake and malabsoption is the most common cause of anemia in gastrectomized patients. This study evaluated the efficacy of administering intravenous iron sucrose for treating patients with anemia. MATERIALS AND METHODS: From May 2007 to October 2007 at Yonsei University Severance Hospital, we reviewed 47 outpatients whose hemoglobin levels were below 11 g/dl after gastrectomy. Iron sucrose was used for iron supplementation. To determine the difference between before and after the treatment (at 1 week, 2 weeks, 3 weeks, 3 months and 6 months after treatment), we prospectively examined such anemia parameters as the hemoglobin level (Hgb), the hematocrit (Hct), serum iron, TIBC, ferritin and transferin. RESULTS: Out of the 47 patients, only 36 completed their treatment. Eleven were male and 25 were female. The Hgb levels, which indicate anemia, were elevated 0.6, 0.8, 1.3, 2.1 and 2.2 g/dl after 1 week, 2 weeks, 3 weeks, 3 months and 6 months after administration, respectively (P<0.001). The changes from 1 week to 3 months were statistically significant, but those from 3 to 6 months were not. The Hgb levels of 26 patients, which accounted for 72% of all the patients, elevated up to 12 g/dl or more. No side effects or complications were found, but there was one case of temporary nausea. CONCLUSION: Anemia after gastrectomy is safely treated in a relatively short time with administering iron sucrose. The patients' Hgb levels are expected to increase in a week and keep increasing up to 3 months.

99mTc-Glucarato como posible trazador de lesiones de cáncer broncopulmonar / 99mTc-Glucarate as a potential novel tracer of lung cancer lesions

El cáncer broncopulmonar (CBP) es la primera causa de muerte por cáncer en el mundo. El CBP a células no pequeñas (CBPCNP) representa el 80 por ciento de la patología neoplásica pulmonar, presentándose frecuentemente al diagnóstico como localmente avanzando o metastásico. Dos tercios se presentan en etapa avanzada, inoperable, integrando la quimioterapia (QT) el tratamiento. Responden favorablemente 30 por ciento de dichos pacientes; los restantes recibirán agentes antineoplásicos sin beneficio terapéutico pero con sus efectos adversos. Ante dicha situación sería de gran relevancia clínica poder contar con una técnica funcional que permitiera evaluar la respuesta a la terapia oncoespecífica. Por tal motivo, nuestro grupo ha desarrollado un protocolo clínico para investigar la capacidad del 99mTc-Glucarato en la evaluación precoz de la respuesta a la QT en este grupo de pacientes. Para ello, es necesario demostrar previamente que las lesiones primarias y secundarias del CBPCNP pueden acumular este trazador. La presente nota clínica describe por vez primera en la literatura indexada la capacidad del 99mTc-Glucarato como marcador funcional de lesiones primarias y metastásicas del CBPCNP.

Calcium glucarate prevents tumor formation in mouse skin / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Calcium Glucarate (Cag), Ca salt of D-glucaric acid is a naturally occurring non-toxic compound present in fruits, vegetables and seeds of some plants, and suppress tumor growth in different models. Due to lack of knowledge about its mode of action its uses are limited in cancer chemotherapy thus the objective of the study was to study the mechanism of action of Cag on mouse skin tumorigenesis.</p><p><b>METHODS</b>We have estimated effect of Cag on DMBA induced mouse skin tumor development following complete carcinogenesis protocol. We measured, epidermal transglutaminase activity (TG), a marker of cell differentiation after DMBA and/or Cag treatment and [3H] thymidine incorporation into DNA as a marker for cell proliferation.</p><p><b>RESULTS</b>Topical application of Cag suppressed the DMBA induced mouse skin tumor development. Topical application of Cag significantly modifies the critical events of proliferation and differentiation TG activity was found to be reduced after DMBA treatment. Reduction of the TG activity was dependent on the dose of DMBA and duration of DMBA exposure. Topical application of Cag significantly alleviated DMBA induced inhibition of TG. DMBA also caused stimulation of DNA synthesis in epidermis, which was inhibited by Cag.</p><p><b>CONCLUSION</b>Cag inhibits DMBA induced mouse skin tumor development. Since stimulation of DNA synthesis reflects proliferation and induction of TG represents differentiation, the antitumorigenic effect of Cag is considered to be possibly due to stimulation of differentiation and suppression of proliferation.</p>

Hepatic microsomal enzyme induction and urinary D-glucaric acid excretion in man

Liver biopsy material of six patients with Hodgkin's disease were used as controls for measurement of drug-metabolizing enzymes. Two patients were treated with 90mg of phenobarbitone to induce the hepatic enzymes and the excretion of D-glucaric acid in urine. The two phenobarbitone-treated patients showed increased activities of hexobarbitone oxidase, cytochrome P-450 and increased microsomal protein content. There was no change in the activities of ethylmorphine-N-demethylase, 1-leucyl-beta-naphthylamide splitting enzyme and UDP-glucuronyle transferase, while p-nitroreductase was repressed. The urinary excretion of D-glucaric acid was increased 2- and 20-fold in the two patients after treatment with phenobarbitone. It is not possible to draw any conclusions from the results regarding the relationship between the in vitro hepatic enzyme activities and the in vivo excretion of D-glucaric acid in the urine due to the small number of cases studied. However the limited observations suggest some interesting possibilities

El ácido D- glucárico urinario y la gamma-glutamiltransferasa sérica como índices de inducción enzimática hepatocitaria en pacientes tratados con fármacos antiepilépticos / Urinary D- glucaric acid and serun gamma-glutamiltransferase as indices of hepatic enzyme induction in patients treated with antiepileptic drugs

Distintos autores han destacado la utilidad del ácido D-glucárico urinario y de la gama-glutamiltransferasa (GGT) sérica en la evaluación de la inducción enzimática hepatocitaria producida por fármacos. En un grupo de pacientes de ambos sexos, tratados con fenobarbital, difenilhidantoína y, ocasionalmente, con carbamazepina y primidona, no se encontró una correlación significativa entre ambas variables bioquímicas (N= 40 , r = 0,238, 0,2> p> 0,1). En 37 pacientes a los que no se había cambinado las pautas posológicas al menos desde hacía un mes, se encontró una correlación significativa entre las dosis totales de los fármacos administrados (expresadas en unidades/día) y los niveles de ácido D-glucárico (r = 0,585, p < 0,001) y de GGT (r= 0,422, p = 0,01). Al hacer una dicotomía por sexos, se encontró una correlación significativa del ácido D-glucárico con la GGT en los hombres (N = 18, r = 0,602, 0,02 > p > 0,01), pero no en las mujeres (N = 19, r = 0,248, 0,4 > p > 0,3). Estos resultados podrían estar relacionados con el efecto depresor de los estrógenos sobre la actividad de la GGT sérica