Association of maternal MTHFD1 and MTHFD2 gene polymorphisms with congenital heart disease in offspring / 中国当代儿科杂志

OBJECTIVES@#To study the association of maternal methylenetetrahydrofolate dehydrogenase 1 (MTHFD1) and methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) gene polymorphisms with congenital heart disease (CHD) in offspring.@*METHODS@#A hospital-based case-control study was conducted. The mothers of 683 children with CHD alone who attended Hunan Children's Hospital, from November 2017 to March 2020 were enrolled as the case group, and the mothers of 740 healthy children who attended the same hospital during the same period and did not have any deformity were enrolled as the control group. A questionnaire survey was performed to collect related exposure data, and then venous blood samples (5 mL) were collected from the mothers to detect MTHFD1 and MTHFD2 gene polymorphisms. A multivariate logistic regression analysis was used to evaluate the association of MTHFD1 and MTHFD2 gene polymorphisms with CHD. The four-gamete test in Haploview 4.2 software was used to construct haplotypes and evaluate the association between haplotypes and CHD. The generalized multifactor dimensionality reduction method and logistic regression analysis were used to examine gene-gene interaction and its association with CHD.@*RESULTS@#The multivariate logistic regression analysis showed that maternal MTHFD1 gene polymorphisms at rs11849530 (GA vs AA: OR=1.49; GG vs AA: OR=2.04) andat rs1256142 (GA vs GG: OR=2.34; AA vs GG: OR=3.25) significantly increased the risk of CHD in offspring (P<0.05), while maternal MTHFD1 gene polymorphisms at rs1950902 (AA vs GG: OR=0.57) and MTHFD2 gene polymorphisms at rs1095966 (CA vs CC: OR=0.68) significantly reduced the risk of CHD in offspring (P<0.05). The haplotypes of G-G-G (OR=1.86) and G-A-G (OR=1.35) in mothers significantly increased the risk of CHD in offspring (P<0.05). The gene-gene interaction analyses showed that the first-order interaction between MTHFD1 rs1950902 and MTHFD1 rs2236222 and the second-order interaction involving MTHFD1 rs1950902, MTHFD1 rs1256142, and MTHFD2 rs1095966 might be associated with risk of CHD (P<0.05).@*CONCLUSIONS@#Maternal MTHFD1 and MTHFD2 gene polymorphisms and their haplotypes, as well as the interaction between MTHFD1 rs1950902 and MTHFD1 rs2236222 and between MTHFD1 rs1950902, MTHFD1 rs1256142, and MTHFD2 rs1095966, are associated with the risk of CHD in offspring.

A high-throughput screening assay for distinguishing nitrile hydratases from nitrilases

A modified colorimetric high-throughput screen based on pH changes combined with an amidase inhibitor capable of distinguishing between nitrilases and nitrile hydratases. This enzymatic screening is based on a binary response and is suitable for the first step of hierarchical screening projects.

Screening, identification and culture optimization of a newly isolated aromatic nitrilase-producing bacterium--Pseudomonas putida CGMCC3830 / 生物工程学报

Microbial nitrilases have attracted increasing attention in nitrile hydrolysis for carboxylic acid production in recent years. A bacterium with nitrilase activity was isolated and identified as Pseudomonas putida CGMCC3830 based on its morphology, physiological and biochemical characteristics, as well as 16S rRNA gene sequence. The nitrilase production was optimized by varying culture conditions using the one-factor-at-a-time method and response surface methodology. Glycerol 13.54 g/L, tryptone 11.59 g/L, yeast extract 5.21 g/L, KH2PO4 1 g/L, NaCl 1 g/L, urea 1 g/L, initial pH 6.0 and culture temperature 30 degrees C were proved to be the optimal culture conditions. It resulted in the maximal nitrilase production of 36.12 U/mL from 2.02 U/mL. Investigations on substrate specificity demonstrate P. putida nitrilase preferentially hydrolyze aromatic nitriles. When applied in nicotinic acid synthesis, 2 mg/mL P. putida cells completely hydrolyzed 20.8 g/L 3-cyanopyridine into nicotinic acid in 90 min. The results indicated P. putida CGMCC3830 displayed potential for industrial production of nicotinic acid.

Biocatalytic desymmetric hydrolysis of 3-(4-chlorophenyl)-glutaronitrile to the key precursor of optically pure baclofen / 生物工程学报

We produced (S)-4-cyano-3-(4-chlorophenyl)-butyrate by highly stereoselective biocatalyst in this study. A nitrilase-producing strain, named Gibberella intermedia WX12, was isolated by 3-(4-chlorophenyl)-glutaronitrile as substrate in the screening with phenol-sodium hypochlorite method. The fermentation conditions and catalytic properties of this strain were investigated. The preferred carbon and nitrogen sources for nitrilase production were lactose (30 g/L) and peptone (20 g/L). After being cultivated for 96 h, the cells were collected for use in biotransformation. The hydrolysis of 3-(4-chlorophenyl)-glutaronitrile was performed at 30 degrees C in phosphate buffer (pH 8.0, 50 mmol/L) for 24 h to give (S)-4-cyano-3-(4-chlorophenyl)-butyric acid with 90% yield and > 99% of ee, which can be used for the synthesis of (R)- and (S)-baclofen. The configuration of product was determined by chemically converting it to baclofen and comparison with the authentic sample by chiral HPLC analysis.

Response surface methodology of nitrilase production by recombinant Escherichia coli

Growth and nitrilase production by recombinant Escherichia coli cells harbouring pET 21 (b) plasmid, for the expression of Pseudomonas putida nitrilase were improved using response surface methodology. Central composite design was used for obtaining ideal concentration of critical medium components which include fructose, tryptone, yeast extract and lactose. The optimal values for the concentration of fructose, tryptone, yeast extract and lactose were found to be 1.13, 2.26, 3.25 and 0.9 percent (w/v), respectively. Here, fructose served as carbon source for the growth while lactose was preferably used as inducer for the expression of foreign protein. Yeast extract in the medium was used as a growth promoter while tryptone was added as a major nitrogen source. Using this optimized medium, an experimental growth of 6.67 (OD at 600 nm) and nitrilase activity of 27.13 U/ml was achieved. This approach for medium development led to an enhancement of the growth and enzyme activity by 1.4 and 2.2 times, respectively, as compared to the un-optimized medium.

Applications of nitrile converting enzymes in the production of fine chemicals / 生物工程学报

Nitriles are an important type of synthetic intermediates in the production of fine chemicals because of their easy preparations and versatile transformations. The traditional chemical conversion of nitriles to carboxylic acids and amides is feasible but it requires relatively harsh conditions of heat, acid or alkali. Nitrile converting enzymes (nitrilase, nitrile hydratase and amidase) which are used as biocatalyst for the production of fine chemicals have attracted substantial interest because of their ability to convert readily available nitriles into the corresponding higher value amides or acids under mild conditions with excellent chemo-, regio- and stereo-selectivities. Many nitrile converting enzymes have been explored and widely used for the production of fine chemicals. In this paper, various examples of biocatalytic synthesis of pharmaceuticals and their intermediates, agrochemicals and their intermediates, food and feed additives, and other fine chemicals are presented. In the near future, an increasing number of novel nitrile converting enzymes will be screened and their potential in the production of useful fine chemicals will be further exploited.

Adenosina deaminasa: su respuesta ante desequilibrios nutricionales / Adenosine deaminase in malnutrition

It has been previously shown that nutritional produces an increment in the activity of adenosine deaminase (ADA) in thymus, serum and other fluids in immunonocompromised patients. This study analyzed if ADA activity could be used as a biochemical indicator of nutritional status in populations at nutritional risk. Twenty six women with anorexia nervosa (AN) 14-32 years old, and 33 obese children (O) 5-13 years old were studied. ADA activity was determined as described by Giusty and Galante, comparing results with control subjects of the same age. The results (x +/- SD, expressed as IU/L) were AN: 25.7+8.2 and 21.0 +/- 4.6; and 0:27.1+9.1° and 23.0±5.6 in experimental and control individuals, respectively (p<0.02). These findings support the hypothesis that ADA activity in serum can be used as a functional indicator related to the defence mechanism in nutritional studies.

Se ha demostrado que el estrés nutricional provoca incremento en la actividad de adenosina deaminasa en timo de rata y además aumenta su actividad en el suero y otros fluidos biológicos de pacientes inmunocompro-metidos. En este estudio se analizó si la determinación de la actividad de ADA en suero podría considerarse parámetro bioquímico funcional en el seguimiento de poblaciones en riesgo nutricional. Se estudiaron mujeres con anorexia nerviosa (AN, 14-32 años) y niños obesos de ambos sexos (O, 5-13 años). La actividad de ADA se determinó por el método de Giusti y Galante. Los resultados se compararon con controles de igual edad. Los resultados (X +/- DE (UI/L)) fueron AN: 25.7 +/- 8.2 vs. 21.0 +/- 4.6 y O: 27.1 +/- 9.1vs. 23.0 +/- 5.6 (p<0.02). Estos hallazgos, avalarían la hipótesis surgida de estudios previos, de proponer la determinación de la actividad sérica de ADA, como un indicador funcional relacionado con los mecanismos de defensa en los estudios de nutrición.

Clinical Features of 212 Cases of Scrub Typhus in Southern Region of Gyeonggi-Do and The Significance of Initial Simple Chest X-Ray / 감염과화학요법

BACKGROUND: Scrub typhus may cause complications such as pneumonia, meningoencephalitis, liver failure, and renal failure, although most of patients with scrub typhus are treated with appropriate antibiotics. Scrub typhus has varying degree of disease severity, from mild to fatal, therefore, early recognition of the clinical parameters representing the severity of the disease is a very important factor for effective treatment. MATERIALS AND METHODS: We investigated clinical, laboratory results and simple chest X-ray images retrospectively by reviewing the medical records of 212 scrub typhus patients, admitted to Catholic University St. Vincent's Hospital from September 2000 to November 2004. RESULTS: Patients demographics revealed a mean age of 59.7, (age: 18 to 90), and 145 of the 212 patients were female (68.4%). Cure was achieved in 211 (99.5%) of the 212 patients with the 1-week course of daily 200 mg doses of doxycycline. Fifty-nine (27.8%) of the patients revealed the abnormal findings, such as interstitial, pneumonic infiltrations, or pleural effusions, in the results of simple chest X-ray images, and cure was achieved in 58 (98.3%) of 59 patients. The median time to defervescence was 27.1 h for the group showing abnormal simple chest X-ray findings, 23.3 h for the group showing normal chest X-ray findings. The titers of serum C-reactive protein and adenine deaminase were significantly higher, and initial hemoglobin and serum albumin levels were lower, in the group with abnormal simple chest X-ray findings, compared to the group with normal simple chest X-ray findings. CONCLUSION: The 1-week course of daily 200-mg doses of doxycycline was effective for the treatment of scrub typhus. Initial simple chest X-ray findings may be useful in the selection of patients, who might show more favorable laboratory findings.

Clinical Features of 212 Cases of Scrub Typhus in Southern Region of Gyeonggi-Do and The Significance of Initial Simple Chest X-Ray / 감염과화학요법

BACKGROUND: Scrub typhus may cause complications such as pneumonia, meningoencephalitis, liver failure, and renal failure, although most of patients with scrub typhus are treated with appropriate antibiotics. Scrub typhus has varying degree of disease severity, from mild to fatal, therefore, early recognition of the clinical parameters representing the severity of the disease is a very important factor for effective treatment. MATERIALS AND METHODS: We investigated clinical, laboratory results and simple chest X-ray images retrospectively by reviewing the medical records of 212 scrub typhus patients, admitted to Catholic University St. Vincent's Hospital from September 2000 to November 2004. RESULTS: Patients demographics revealed a mean age of 59.7, (age: 18 to 90), and 145 of the 212 patients were female (68.4%). Cure was achieved in 211 (99.5%) of the 212 patients with the 1-week course of daily 200 mg doses of doxycycline. Fifty-nine (27.8%) of the patients revealed the abnormal findings, such as interstitial, pneumonic infiltrations, or pleural effusions, in the results of simple chest X-ray images, and cure was achieved in 58 (98.3%) of 59 patients. The median time to defervescence was 27.1 h for the group showing abnormal simple chest X-ray findings, 23.3 h for the group showing normal chest X-ray findings. The titers of serum C-reactive protein and adenine deaminase were significantly higher, and initial hemoglobin and serum albumin levels were lower, in the group with abnormal simple chest X-ray findings, compared to the group with normal simple chest X-ray findings. CONCLUSION: The 1-week course of daily 200-mg doses of doxycycline was effective for the treatment of scrub typhus. Initial simple chest X-ray findings may be useful in the selection of patients, who might show more favorable laboratory findings.

Expression of Vitreoscilla hemoglobin and nitrilase in the yeast Pichia pastoris / 生物工程学报

The expression of the vgb gene in vivo could improve the fermentation density and then contribute the extracellular secretion of the product of bxn gene. Constructed the recombination plasmid pPIC9K-vgbbxn and transformed into Pichia pastoris GS115. The results of PCR and SDS-PAGE indicate that the vgb gene and bxn gene had integrated into the genome of Pichia pastoris GS115 and expressed in efficient level. Also, the protein activity of their products had been verified respectively. Shake flask fermentation experiments showed that the presence of VHb in yeast Pichia pastoris efficiently enhanced cell growth and secretive expression of bxn gene under hypoxic habitats.