Advances in the microbial production of the compatible solute ectoine: a review / 生物工程学报

Ectoine is an amino acid derivative and an important natural product in halophilic microorganisms. It plays an important role in protecting cells and stabilizing biological macromolecules, and can be widely used in biomedical fields such as drug preparation adjuvants, organ transplantation and preservation, skin wound repair and cosmetics. Due to the medical value and commercial market demand of ectoine, this article summarized the recent advances in the microbial production of ectoine, including the mutation and breeding of hyper-producing strains, construction of genetically and metabolically engineered strains, optimization of fermentation processes, and extraction and purification processes. The application of multi-omics technologies and computational biology to develop an ectoine producing cell factory was prospected, with the aim to provide a reference for ectoine overproduction.

Fingerprinting analysis of non-saponins from water-soluble part and determination of dencichine from Panax notoginseng, P. ginseng and P. quinquefolium based on liquid chromatography coupled with charged aerosol detection / 中国中药杂志

This work describes the holistic fingerprinting method based on liquid chromatography coupled with charged aerosol detection(CAD) to profile non-saponin from water-soluble parts and determination of dencichine in Panax ginseng(PG), P. quinquefolium(PQ) and P. notoginseng(PNG). Sample extraction was carried out by water with ultra sonication for 30 min, which was eluted by Retain PEP for further analysis. The analysis was performed on a Hypercarb of porous graphitized carbon(3.0 mm×150 mm, 3 μm) column with acetonitrile and 0.1% perfluoropentanoic acid as mobile phase at a flow rate of 0.8 mL·min~(-1). Temperature of evaporator and nitrogen pressure for CAD were set at 50 ℃and 60.1 psi(1 psi≈6.895 kPa), respectively. As a result, dencichine and other polar components had a good performance on resolution and retention. The correlation coefficient(R~2) of dencichine was 0.998 2 in the concentration from 0.019 2 to 0.48 μg·mL~(-1). Limit of quantitation calculated by signal to noise of 10 was 7.4 ng·mL~(-1), and the recovery ranged from 95.52% to 102.7%. Chemical profile of the water-soluble part from PG, PQ and PNG was similar holistically, while the relative content for dencichine and other partial components varied significantly. The proposed method was used for characteristic of chemical profiling for non-saponin from water-soluble part, and determination of dencichine in PG, PQ and PNG.

Genotyping of Low β -ODAP Grass Pea (Lathyrus sativus L. ) Germplasm with EST-SSR Markers

Abstract Grass pea (Lathyrus sativus L.) is an important protein source in arid regions as both human and animal food. Despite its significance, the use of grass pea is limited by the presence of β-N-oxalyl-L-a,b-diaminopropionic acid (β-ODAP) which can cause neurological disorders. Breeding studies in grass pea have therefore focused on developing high-yielding varieties with low β-ODAP content. However, the narrow range of genetic diversity and the restricted genomic tools in grass pea have slowed progress in such breeding. The present investigation was conducted to explore the genetic diversity of low β-ODAP germplasm consisting of 22 accessions with 31 EST-SSR markers. The molecular analyses revealed a total of 133 alleles ranging from 142 to 330 bp with a mean number of alleles per locus of 4.29. The mean polymorphic information content (PIC) value was calculated as 0.49, and the EST-SSRs in loci S5, S6 and S116 were of the most informative PICs. A dendrogram based on Nei's genetic distance matrix revealed that breeding lines were grouped in two main clusters. Genetic distances were higher between GP6/GP11, GP4/GP11 and GP5/GP8 accessions which could be further used in crop improvement studies for developing wider genetic diversity.

Genotype and Phenotype Analysis in Pediatric Patients with Cystinuria

Cystinuria is an inherited disorder characterized by defective renal reabsorption of cystine and dibasic amino acids leading to nephrolithiasis. This study was conducted to analyze the genotypes and phenotypes of pediatric patients with cystinuria. Eight children from Seoul National University Hospital and Asan Medical Center presenting with cystinuria from January 2003 to June 2016 were retrospectively analyzed. Mutational studies were performed by direct sequencing. Two of the 8 were male and 6 were female. The median ages at onset and diagnosis were 1.5 (range, 0.3–13.6) and 2.6 (range, 0.7–16.7) years, respectively. The median followed up was 7.7 (range, 3.4–14.0) years. Mutational analyses were performed in 7 patients and revealed biallelic SLC3A1 mutations (AA genotype) in 4 patients, a single heterozygous SLC3A1 mutation (A- genotype) in 1 patient, biallelic SLC7A9 mutations (BB genotype) in 1 patient, and a single heterozygous SLC7A9 mutation (B- genotype) in 1 patient. Two of the mutations were novel. No genotype-phenotype correlations were observed, except for earlier onset age in patients with non-AA genotypes than in patients with the AA genotype. All patients suffered from recurrent attacks of symptomatic nephrolithiasis, which lead to urologic interventions. At the last follow-up, 3 patients had a mild-to-moderate degree of renal dysfunction. This is the first study of genotypic and phenotypic analyses of patients with cystinuria in Korea.

Content determination of dencichine in Panax Notoginseng by a reversed phase ion-pair chromatography / 中国中药杂志

To build a reversed phase ion-pair chromatography to determination content of Dencichine from Panax notoginseng. Using Tetrabutyl ammonium hydroxide ions by the combination of reagent and HPLC method without derivatization to test the content of dencichine directly. The optimum conditions of supersonic extraction were solid-to-liquid ratio 1: 20, Continuous ultrasonic extraction: twice, each time 15 minutes; 3,500 r · min⁻¹, then centrifuging 15 minutes. Dencichine in different age, place, part and the different Processing mode were examined. The method is simple with sound separation degree and stability, which can facilitate the determination of dencichine content directly and provide the basis in quality standard of raw material.

Determination of dencichine in Sanqi tablet by HILIC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HILIC method for determination of dencichine in Sanqi tablet and evaluate the quality of Sanqi tablet of different hatches from various manufactures in the market.</p><p><b>METHOD</b>The chromatographic separation was conducted on a Thermo HILIC column (4.6 mm x 250 mm, 5 microm) kept at 25 degrees C with acetonitrile and 0.1% H3PO4 (60:40) as the mobile phase. The flow rate was set at 1 mL x min(-1) and the detection wavelength was set at 213 nm.</p><p><b>RESULT</b>The contents of dencichine in Sanqi tablet ranged from 1.60 to 4.31 mg x g(-1).</p><p><b>CONCLUSION</b>The well established method was successfully applied to determine dencichine in Sanqi tablet. The results demonstrated that this method was simple, accurate and could be applied for quality control of Sanqi as well as its associated preparations.</p>

A Case of Cystinuria with Multiple Renal Stones in an 8-month-old Girl

Cystinuria is an autosomal recessive disease characterized by impaired transport of cystine and dibasic amino acids in the proximal renal tubule, resulting in the formation of cystine stones. It is believed to account for about 1% of all kidney stones and up to 10% of pediatric stones. Here we report a case of cystinuria with multiple renal stones confirmed by genetic mutational analysis. An 8-month-old girl was admitted to AMC with persistent fever and multiple renal stones. A renal sonogram showed multiple stones at the right renal pelvis, right distal ureter, and left renal medullary portion. An approximately 1 cm renal stone was extracted spontaneously, and stone analysis revealed it to be composed entirely of cystine. Cystinuria was confirmed by increased urine dibasic amino acid levels, including cysteine, and genetic mutational analysis showed the patient to be a homozygote for the pathogenic c. 1820del (p.L607fs) of SLC3A1. Despite treatment with oral hydration and urinary alkalinization, and restricted intake of animal protein, the stones increased in size and number. The patient has since been treated with tiopronin.

Experimental study of the anti-inflammatory and analgesic effects of diethyl 1,3-dicyclohexyl- 1,2,3,6-tetrahydropyrimidine-4,5-dicarboxylate / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the anti-inflammatory and analgesic activities of diethyl 1,3-dicyclohexyl-1,2,3,6-tetrahydropyrimidine-4,5-dicarboxylate (ZL-5010) in vivo and in vitro.</p><p><b>METHODS</b>The analgesic effect of ZL-5010 was evaluated by acetic acid-induced writhing response in mice, and the anti-inflammatory effects was assessed in mice with xylene-induced ear edema and in rats with carrageenan-induced paw edema. Mouse peritoneal exudate cells activated by bacterial lipopolysaccharides (LPS) were used to evaluate the anti-inflammatory effect of ZL-5010 in vitro. The levels of interleukin-1β (IL -1β) and tumor necrosis factor-α (TNF-α) in the cell culture supernatant were measured using enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>At the doses of 0.25 and 0.5 mmol/kg, ZL-5010 administered by gavage once daily for 3 days significantly reduced acetic acid-induced writhing frequency and suppressed xylene-induced ear edema in mice, and alleviated paw edema induced by carrageenan in rats (P<0.05). The agent also inhibited the production of the pro-inflammatory cytokines IL-1β and TNF-α by LPS-induced mouse peritoneal exudate cells in vitro, with the statistically significant minimum effective concentrations of 10 and 20 µmol/L, respectively (P<0.05).</p><p><b>CONCLUSION</b>ZL-5010 administered by gavage has anti-inflammatory and analgesic effects in mice and rats, and in mouse peritoneal exudate cell cultures, the agent also inhibits the production of the pro-inflammatory cytokines IL-1β and TNF-α.</p>

Effects of fluvastatin on the activation of p38 mitogen-activated protein kinase in glomerular mesangial cells under high concentration of glucose / 药学学报

This study is to investigate the effects of fluvastatin on the activation of p38 mitogen-activated protein kinase (p38 MAPK) and cAMP response element-binding protein (CREB1) in glomerular mesangial cells under high concentration of glucose. High concentration glucose and fluvastatin were used to stimulate the cultured rat glomerular mesangial cells (GMCs) in vitro. The protein expressions of p38 MAPK, CREB1, p-p38 MAPK and p-CREB1 were observed with Western blotting. TGF-beta1 and fibronectin (FN) mRNA were measured with reverse transcription and polymerase chain reaction (RT-PCR). The protein synthesis of laminine (LN) and type IV collagen in the supernatants of the GMCs were detected with radioimmunoassay. Compared with low glucose control group, the expressions of p-p38 MAPK, p-CREB1 were increased obviously in high glucose group, TGF-beta1 mRNA and FN mRNA, LN and type IV collagen in the supernatants were increased significantly in GMCs under high concentration glucose medium. The expression levels of p-p38 MAPK, p-CREB1, TGF-beta1 mRNA, and FN mRNA, LN and type IV collagen in the supernatants were significantly lower in the fluvastatin group than those in the high concentration glucose group. It is concluded that fluvastatin can inhibit over production of TGF-beta1 and ECM proteins in GMCs under high concentration of glucose, partly by regulating the phosphorylation of p38 MAPK and CREB1.

The adaptor protein LAD/TSAd mediates laminin-dependent T cell migration via association with the 67 kDa laminin binding protein

The adaptor protein, LAD/TSAd, plays essential roles in T cell activation. To further understand the functions of this protein, we performed yeast two-hybrid screening using TSAd as bait and identified 67 kDa laminin binding protein (LBP) as the interacting partner. Subsequently, TSAd-LBP interaction was confirmed in D1.1 T cell line. Upon costimulation by T cell receptor (TCR) plus laminin crosslinking or TCR plus integrin alpha6 crosslinking, LBP was coimmunoprecipitated with TSAd. Moreover, TCR plus laminin costimulation-dependent T cell migration was enhanced in D1.1 T cells overexpressing TSAd but was disrupted in D1.1 cells overexpressing dominant negative form of TSAd or TSAd shRNA. These data show that, upon TCR plus integrin costimulation, TSAd associates with LBP and mediates T lymphocyte migration.

Cloning and characterization of ectABC cluster from Bacillus alcalophilus DTY1 / 生物工程学报

Bacillus alcalophilus DTY1, one moderate halophytic bacterium isolated from saline soil in Loess Plateau of China, was characterized with efficient production of ectoine. In this study, the gene cluster ectABC taking in charge of biosynthesizing ectoine was cloned from the genomic library of strain DTY1. Nucleotide sequencing indicated that ectA, ectB and ectC were predicted to encode peptides of 169, 428 and 132 amino acids, respectively. The deduced amino acid sequences of EctA, EctB and EctC share 59%, 81% and 81% identity to 2,4-diaminobutyric acid acetyltransferase, 2,4-diaminobutyric acid transaminase and ectoine synthase of B. halodurans C-125, respectively. A fragment containing ectABC genes was introduced into B. cereus Z, which made the transgenic Z cells increased tolerance to salt, remarkably. HPLC analysis of ectoine in the transgenic Z cells revealed that 70.1 mg/g ectoine was detected in 1.0% NaCl medium and 118.6 mg/g ectoine in 5.0% NaCl medium. Furthermore, as the concentration of salt increased, transgenic Z cells accumulated more ectoine. These results suggest that ectoine is an important facet in B. alcalophilus DTY1 to high-osmolarity surroundings, and the expression of ectABC is induced by salt strength.

Analysis of dencichine by HPLC with pre-column derivatization / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a reversed-phase high performance liquid chromatorgraphy (RP-HPLC) method for detecting the dencichine in Panax notoginseng extracts and drug preparations.</p><p><b>METHOD</b>Dencichine was extracted with the borate buffer (pH 9. 18) and the clear supernatant was used for the derivatization. Pre-column derivatization was performed using 9-fluorenylmethyl chloroformate (FMOC) to form derivatives. The mobile phase consisted of methanol and 0. 05 mol x L( -1) NaH2 PO4 (48: 52) (pH adjusted to 7.4 with NaOH solution) in a flow rate of 1.0 mL m min(-1). The ultraviolet (UV) detection wavelength was set at 262 nm.</p><p><b>RESULT</b>The linearity was demonstrated over a wide range of concentration from 1.76 mg L(-1) to 352 mg x L(-1) for dencichine. The detection limit was determined to be 60 microg x L(-1). The derivative was stable and the derivatization agent did not influence the measurement of dencichine. The average recovery rate was 95. 3% and the relative standard derivation (RSD) was 1. 7%. The method was used to determine dencichine in different P. notoginseng extracts and drug preparations.</p><p><b>CONCLUSION</b>This method is simple, fast and sensitive, suitable for determining the dencichine in P. notoginseng extracts and drug preparations as well as for the study of the dencichine metabolism in vivo.</p>

Diamino fluoreto de prata: Recurso subestimado em Saúde Pública? / Diammine silver fluoride: is its use underestimated in Public Dental Health?

O diamino fluoreto de prata tem sido um recurso pouco utilizado em saúde pública. No entanto, a análise do seu mecanismo de ação e principais propriedades demonstradas nos estudos publicados permite concluir que essa substância tem ampla aplicação no serviço público ou privado. Ressalta-se que a escolha desses produtos poderia ser feita em determinado momento (fase aguda da doença), segundo a necessidade dos pacientes (especiais, não colaboradores) e de acordo com as condições de trabalho (ausência de outros materiais). Esse tratamento não é necessariamente definivo. Aliado a medidas educativas/preventivas até poderia ser o único tratamento, mas não há qualquer impedimento de recuperar a anatomia e a estética dentária, quando houver esta possibilidade.

Synthesis of some N,N- [4,4-carbonyldiphthaloyl] diamino acids and dipeptide derivatives and studies of their antimicrobial activities and the effect of gamma rays on some of these compounds

The synthesis of different N, N'- [4,4-carbonyldiphthaloyl] diamino acids [II-IX] and some of their corresponding acid chlorides [X-XII] and methyl esters [XIII-XV] are described. Coupling of N, N'-[4,4- carbonyldiphthaloyl] diamino acids with amino acid methyl ester hydrochlorides in dioxan containing triethyl amino using the dicyclohexyl-carbodiimide [DCC] method furnishes the desired N, N'- [4,4'-carbonyldiphthaloyl] bis-dipeptide methyl esters [XVI-XXIV]. Most of the synthesized N, N'-[4,4'-carbonyldiphthaloyl] diamino acids, esters and bis-dipeptide methyl esters were found to be active against number of microorganisms. The effect of gamma irradiation on some of these compounds was studied

Effect of the boiling and decanting method of Khesari (Lathyruss sativus) detoxification, on changes in selected nutrients

Es un hecho más que establecido que la leguminosa conocida como Khesari (Lathyrus sativus) causa latirismo, enfermedad que en los humanos se caracteriza por parálisis de los miembros inferiores. El constituyente tóxico es un aminoácido identificado como "B-Oxalyl-amino L-alanina" (BOAA). Se ha informado que si la leguminosa se hierve durante dos horas, y luego se decanta el agua, se logra eliminar casi el 85% del aminoácido. Esta investigación, por lo tanto, constituye un esfuerzo para prevenir que - en forma simultánea a la eliminación de la toxicidad - ocurran pérdidas de otros nutrientes. En el caso del dahl (descascarados y separados los cotiledones), se ha observado que se pierde casi la mitad de proteínas, 80.36% de azúcares totales, 63.13% de azúcares reductoras, 86.05% de aminoácidos, y toda la tiamina, riboflavina y niacina, mientras que con tan sólo una hora de tratamiento, las pérdidas respectivas de las semillas enteras son de 47.25%, 45.73%, 74.69% y 80.00%, así como todas las vitaminas. Las pérdidas del aminoácido tóxico del dahl y de las semillas enteras son de 71.46% y 68.74% respectivamente. Se describen, asimismo, los datos en cuanto a las pérdidas que courren con el tratamiento de dos y de tres horas