Anti-inflammatory and antioxidant properties of betaine protect against sepsis-induced acute lung injury: CT and histological evidence

The aim of this research was to determine the anti-inflammatory effect of betaine on sepsis-induced acute respiratory distress syndrome (ARDS) in rats through histopathological examination, radiologic imaging, and biochemical analysis. Eight rats were included in the control group, and no procedure was performed. Feces intraperitoneal procedure (FIP) was performed on 24 rats to create a sepsis-induced ARDS model. These rats were separated into three groups as follows: FIP alone (sepsis group, n=8), FIP + saline (1 mL/kg, placebo group, n=8), and FIP + betaine (500 mg/kg, n=8). Computed tomography (CT) was performed after FIP, and the Hounsfield units (HU) value of the lungs was measured. The plasma levels of tumor necrosis factor (TNF)-α, interleukin-1β (IL-1β), IL-6, C-reactive protein, malondialdehyde (MDA), and lactic acid (LA) were determined, and arterial oxygen pressure (PaO2) and arterial CO2 pressure (PaCO2) were measured from an arterial blood sample. Histopathology was used to evaluate lung damage. This study completed all histopathological and biochemical evaluations in 3 months. All evaluated biomarkers were decreased in the FIP + betaine group compared to FIP + saline and FIP alone (all P<0.05). Also, the parenchymal density of the rat lung on CT and histopathological scores were increased in FIP + saline and FIP alone compared to control and these findings were reversed by betaine treatment (all P<0.05). Our study demonstrated that betaine suppressed the inflammation and ameliorated acute lung injury in a rat model of sepsis.

Organic osmolyte betaine mitigates the deleterious effects of Diclofenac in vivo in wistar albino rats

Abstract Diclofenac sodium (DF) is a non-steroidal anti-inflammatory drug (NSAID) that possesses antipyretic, analgesic, antinociceptive and anti-inflammatory activities. Like other NSAIDs, DF is known to be associated with renal, cardiovascular, and gastrointestinal complications. The present study was carried out to evaluate the adverse effects of DF in vivo in wistar albino rats and to assess if oral administration of the organic osmolyte betaine mitigates the adverse effect of DF. Eighteen male Wistar rats were divided into three groups, one group of animals was fed orally with 20 mg/kg of DF once/day, and the other group received a combination of 20 mg/kg of DF and 30 mg/kg of betaine, once/day. Apart from the hematological and biochemical parameters, histopathological changes in the liver, lungs, brain, heart and kidney were also investigated. Histopathological alterations that were found in the liver, kidney, and lungs of DF-treated animals were found to be minimal or absent in DF + betaine-treated animals, as compared to untreated control. The results showed that betaine mitigates the adverse effects associated with DF treatment.

Analyzing the difference between atomic fluorescence spectrometry and ICP-MS in the determination of arsenic in urine / 中国职业医学

@#Abstract: Objective （ ） To compare the measured results of arsenic in urine by atomic fluorescence spectrometry AFS and - （ - ）， Methods inductively coupled plasma mass spectroscopy ICP MS and analyze the reasons of the difference. The samples WS/T 474-2015 Determination of Arsenic in Urine by Hydride Generation Atomic Fluorescence were pretreated according to Spectrometry， （ ∶ ∶ ∶∶ ，V/V/V） and digested with mixed acid nitric acid sulfuric acid perchloric acid=3 1 1 and then determined by - - AFS and ICP MS. The samples were diluted with 0.50% nitric acid and determined by ICP MS. The samples included urine ， ， （ arsenic quality control samples inorganic arsenic supplemented samples and organic arsenic arsenic choline and arsenic ） - betaine supplemented samples. Standard curve method was used to compare the results of AFS method and ICP MS method. Results （ ） （ ） The results of quality control samples by AFS method digestion and ICP-MS method without digestion were ， - within the range of reference values but the values obtained by AFS method were lower than those obtained by ICP MS method. - - - ， The recovery of AFS and ICP MS was 97.79% 100.82% and 99.55% 99.98% respectively. In the middle and high ， - （ P ） concentration groups the measured values of inorganic arsenic by AFS were lower than that by ICP MS all <0.01 . The （ ） - recovery of arsenic betaine and arsenic choline by AFS method digestion was only 2.17% 2.63%. The values of arsenic betaine （ ） - （ and arsenic choline measured by AFS method digestion were lower than those measured by ICP MS method without ） - （ ）（ P ）Conclusion digestion and ICP MS method digestion all <0.01 . The result of urine arsenic measured by AFS method - ， was lower than that measured by ICP MS method which may be related to the mixed acid digestion of AFS method. Keywords: ； - ； ； ； ； ；

Antioxidant status and their enhancements strategies for water stress tolerance in chickpea / Status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico

Water stress executes severe influences on the plant growth and development through modifying physio-chemical properties. Therefore, a field experiment was designed to evaluate the antioxidant status and their enhancements strategies for water stress tolerance in chickpea on loam and clay loam soils under agro-ecological conditions of Arid Zone Research Institute, Bahawalpur (29.3871 °N, 71.653 °E) and Cholistan farm near Derawer (28.19°N, 71.80°E) of Southern Punjab, Pakistan during winter 2014-15. Experimental treatments comprised of two chickpea cultivars i.e. Bhakhar 2011 (drought tolerant) and DUSHT (drought sensitive), two water stress levels i.e. water stress at flowering stage and water stress at flowering + pod formation + grain filling stage including well watered (control) and two exogenous application of osmoprotectants i.e. glycine betaine (GB) 20 ppm and proline 10 uM including distilled water (control). Results indicated that water stress at various growth stages adversely affects the growth, yield and quality attributes of both chickpea cultivars. Exogenous application of GB and proline improved the growth, yield and quality parameters of both chickpea cultivars even under water stress conditions. However, superior results were obtained with exogenously applied GB on Bhakhar 2011 under well-watered conditions. Similarly, foliar spray of GB on chickpea cultivar Bhakhar 2011 under stress at flowering + pod formation + grain filling stage produced maximum superoxide dismutase, peroxidase and catalase contents. These results suggested that application of GB mitigates the adverse effects of water stress and enhanced tolerance in chickpea mainly due to higher antioxidant enzymes activity, demonstrating the protective measures of plant cells in stress condition. Hence, antioxidants status might be a suitable method for illustrating water stress tolerance in chickpea.

O estresse hídrico exerce fortes influências no crescimento e no desenvolvimento das plantas, modificando as propriedades físico-químicas. Portanto, a presente atividade de pesquisa foi projetada para avaliar o status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico em condiçõesa groecológicas, no Instituto de Pesquisa da Zona Árida, Bahawalpur (29.3871 ° N, 71.653 ° E) e fazenda do Cholistan, perto de Derawer (28.19 ° N, 71,80 ° E), no sul de Punjab, Paquistão, durante Rabi 2014-15. Tratamentos experimentais compostos de dois genótipos de grão-de-bico, como Bhakhar 2011 (tolerante à seca) e DUSHT (sensível à seca), dois níveis de estresse hídrico, ou seja, estresse hídrico no estágio de floração, estresse hídrico na fase de floração e estresse hídrico na fase de floração + formação de vagem + estágio de enchimento de grãos, incluindo água bem controlada (controle) e duas aplicações exógenas de osmoprotetores, isto é, glicina betaína 20 ppm e prolina 10 uM, incluindo água destilada (controle). Os resultados indicaram que o estresse hídrico em vários estágios de crescimento afeta negativamente os atributos de crescimento, rendimento e qualidade de ambas as cultivares de grão-de-bico. A aplicação exógena de glicina betaína e prolina melhorou os parâmetros de crescimento, rendimento e qualidade de ambos os genótipos de grão- de-bico, mesmo sob condições de estresse hídrico. No entanto, resultados superiores foram obtidos com glicina betaína aplicada exogenamente em Bhakhar 2011, em condições bem regadas. Além disso, o spray foliar de glicina betaína na cultivar de grão-de-bico Bhakhar 2011, sob estresse na fase de floração + formação de vagem + enchimento de grãos, produziu o máximo de superóxido dismutase, peroxidase e catalase. Esses resultados sugeriram que a aplicação de glicina betaína atenua os efeitos adversos do estresse hídrico e aumenta a [...].

Antioxidant status and their enhancements strategies for water stress tolerance in chickpea

Abstract Water stress executes severe influences on the plant growth and development through modifying physio-chemical properties. Therefore, a field experiment was designed to evaluate the antioxidant status and their enhancements strategies for water stress tolerance in chickpea on loam and clay loam soils under agro-ecological conditions of Arid Zone Research Institute, Bahawalpur (29.3871 °N, 71.653 °E) and Cholistan farm near Derawer (28.19°N, 71.80°E) of Southern Punjab, Pakistan during winter 2014-15. Experimental treatments comprised of two chickpea cultivars i.e. Bhakhar 2011 (drought tolerant) and DUSHT (drought sensitive), two water stress levels i.e. water stress at flowering stage and water stress at flowering + pod formation + grain filling stage including well watered (control) and two exogenous application of osmoprotectants i.e. glycine betaine (GB) 20 ppm and proline 10 uM including distilled water (control). Results indicated that water stress at various growth stages adversely affects the growth, yield and quality attributes of both chickpea cultivars. Exogenous application of GB and proline improved the growth, yield and quality parameters of both chickpea cultivars even under water stress conditions. However, superior results were obtained with exogenously applied GB on Bhakhar 2011 under well-watered conditions. Similarly, foliar spray of GB on chickpea cultivar Bhakhar 2011 under stress at flowering + pod formation + grain filling stage produced maximum superoxide dismutase, peroxidase and catalase contents. These results suggested that application of GB mitigates the adverse effects of water stress and enhanced tolerance in chickpea mainly due to higher antioxidant enzymes activity, demonstrating the protective measures of plant cells in stress condition. Hence, antioxidants status might be a suitable method for illustrating water stress tolerance in chickpea.

Resumo O estresse hídrico exerce fortes influências no crescimento e no desenvolvimento das plantas, modificando as propriedades físico-químicas. Portanto, a presente atividade de pesquisa foi projetada para avaliar o status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico em condições agroecológicas, no Instituto de Pesquisa da Zona Árida, Bahawalpur (29.3871 ° N, 71.653 ° E) e fazenda do Cholistan, perto de Derawer (28.19 ° N, 71,80 ° E), no sul de Punjab, Paquistão, durante Rabi 2014-15. Tratamentos experimentais compostos de dois genótipos de grão-de-bico, como Bhakhar 2011 (tolerante à seca) e DUSHT (sensível à seca), dois níveis de estresse hídrico, ou seja, estresse hídrico no estágio de floração, estresse hídrico na fase de floração e estresse hídrico na fase de floração + formação de vagem + estágio de enchimento de grãos, incluindo água bem controlada (controle) e duas aplicações exógenas de osmoprotetores, isto é, glicina betaína 20 ppm e prolina 10 uM, incluindo água destilada (controle). Os resultados indicaram que o estresse hídrico em vários estágios de crescimento afeta negativamente os atributos de crescimento, rendimento e qualidade de ambas as cultivares de grão-de-bico. A aplicação exógena de glicina betaína e prolina melhorou os parâmetros de crescimento, rendimento e qualidade de ambos os genótipos de grão- de-bico, mesmo sob condições de estresse hídrico. No entanto, resultados superiores foram obtidos com glicina betaína aplicada exogenamente em Bhakhar 2011, em condições bem regadas. Além disso, o spray foliar de glicina betaína na cultivar de grão-de-bico Bhakhar 2011, sob estresse na fase de floração + formação de vagem + enchimento de grãos, produziu o máximo de superóxido dismutase, peroxidase e catalase. Esses resultados sugeriram que a aplicação de glicina betaína atenua os efeitos adversos do estresse hídrico e aumenta a tolerância no grão-de- bico, principalmente pela maior atividade de enzimas antioxidantes, demonstrando medidas protetoras das células vegetais em condições de estresse. Portanto, o status de antioxidantes pode ser um método adequado para ilustrar a tolerância ao estresse hídrico no grão-de-bico.

Antioxidant status and their enhancements strategies for water stress tolerance in chickpea / Status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico

Water stress executes severe influences on the plant growth and development through modifying physio-chemical properties. Therefore, a field experiment was designed to evaluate the antioxidant status and their enhancements strategies for water stress tolerance in chickpea on loam and clay loam soils under agro-ecological conditions of Arid Zone Research Institute, Bahawalpur (29.3871 °N, 71.653 °E) and Cholistan farm near Derawer (28.19°N, 71.80°E) of Southern Punjab, Pakistan during winter 2014-15. Experimental treatments comprised of two chickpea cultivars i.e. Bhakhar 2011 (drought tolerant) and DUSHT (drought sensitive), two water stress levels i.e. water stress at flowering stage and water stress at flowering + pod formation + grain filling stage including well watered (control) and two exogenous application of osmoprotectants i.e. glycine betaine (GB) 20 ppm and proline 10 uM including distilled water (control). Results indicated that water stress at various growth stages adversely affects the growth, yield and quality attributes of both chickpea cultivars. Exogenous application of GB and proline improved the growth, yield and quality parameters of both chickpea cultivars even under water stress conditions. However, superior results were obtained with exogenously applied GB on Bhakhar 2011 under well-watered conditions. Similarly, foliar spray of GB on chickpea cultivar Bhakhar 2011 under stress at flowering + pod formation + grain filling stage produced maximum superoxide dismutase, peroxidase and catalase contents. These results suggested that application of GB mitigates the adverse effects of water stress and enhanced tolerance in chickpea mainly due to higher antioxidant enzymes activity, demonstrating the protective measures of plant cells in stress condition. Hence, antioxidants status might be a suitable method for illustrating water stress tolerance in chickpea.

O estresse hídrico exerce fortes influências no crescimento e no desenvolvimento das plantas, modificando as propriedades físico-químicas. Portanto, a presente atividade de pesquisa foi projetada para avaliar o status antioxidante e suas estratégias de aprimoramento para tolerância ao estresse hídrico no grão-de-bico em condições agroecológicas, no Instituto de Pesquisa da Zona Árida, Bahawalpur (29.3871 ° N, 71.653 ° E) e fazenda do Cholistan, perto de Derawer (28.19 ° N, 71,80 ° E), no sul de Punjab, Paquistão, durante Rabi 2014-15. Tratamentos experimentais compostos de dois genótipos de grão-de-bico, como Bhakhar 2011 (tolerante à seca) e DUSHT (sensível à seca), dois níveis de estresse hídrico, ou seja, estresse hídrico no estágio de floração, estresse hídrico na fase de floração e estresse hídrico na fase de floração + formação de vagem + estágio de enchimento de grãos, incluindo água bem controlada (controle) e duas aplicações exógenas de osmoprotetores, isto é, glicina betaína 20 ppm e prolina 10 uM, incluindo água destilada (controle). Os resultados indicaram que o estresse hídrico em vários estágios de crescimento afeta negativamente os atributos de crescimento, rendimento e qualidade de ambas as cultivares de grão-de-bico. A aplicação exógena de glicina betaína e prolina melhorou os parâmetros de crescimento, rendimento e qualidade de ambos os genótipos de grão- de-bico, mesmo sob condições de estresse hídrico. No entanto, resultados superiores foram obtidos com glicina betaína aplicada exogenamente em Bhakhar 2011, em condições bem regadas. Além disso, o spray foliar de glicina betaína na cultivar de grão-de-bico Bhakhar 2011, sob estresse na fase de floração + formação de vagem + enchimento de grãos, produziu o máximo de superóxido dismutase, peroxidase e catalase. Esses resultados sugeriram que a aplicação de glicina betaína atenua os efeitos adversos do estresse hídrico e aumenta a tolerância no grão-de- bico, principalmente pela maior atividade de enzimas antioxidantes, demonstrando medidas protetoras das células vegetais em condições de estresse. Portanto, o status de antioxidantes pode ser um método adequado para ilustrar a tolerância ao estresse hídrico no grão-de-bico.

Dermatite de contato agravada por outra sensibilidade em automedicação / Contact dermatitis aggravated by other sensitivity in self-medication

RESUMO: Dermatite alérgica de contato é uma doença cutânea inflamatória, não infecciosa, cuja base do tratamento é a identificação e eliminação do agente causal. Cocoamidopropil betaína é um surfactante muito utilizado nos produtos de uso pessoal, notadamente de uso capilar. Essa substância não está presente na bateria padrão brasileira. Neomicina é um antibiótico usado em preparações tópicas. Objetivamos mostrar paciente que desenvolveu alergia no couro cabeludo e que fez erroneamente automedicação com produto que continha substância a qual era ainda mais sensível. O caso é de uma mulher, 36 anos, evoluindo há dois meses com eczema pruriginoso, em áreas de implantação capilar e nuca. Relatava progressiva piora. Diante da suspeita de dermatite de contato, foi realizado teste de contato, utilizando-se da bateria padrão Latino-Americana. Com 96 horas (D4) evidenciou-se positividade leve (+) para cocoamidopropil betaína e forte (++) para neomicina. O resultado positivo para a neomicina foi intrigante, uma vez que a observação do rótulo dos produtos de uso pessoal não a continha. Diante do resultado do teste, após ser questionada novamente, ela confirmou a omissão da automedicação diária com pomada de neomicina. Em conclusão, mostramos a alergia a produtos de uso capilar. Reforçamos a necessidade de se fazer um teste de contato com bateria padrão atualizada. Por fim, alertamos sobre o risco da automedicação. (AU)

ABSTRACT: Allergic contact dermatitis is an inflammatory, non-infectious skin disease. The treatment is based on the identification and elimination of the causal agent. Cocamidopropyl betaine is a surfactant widely used in products for personal use, especially capillary use. This substance is not present in the Brazilian baseline series. Neomycin is an antibiotic used in topical preparations. We aimed to show a patient who developed na allergy in the scalp and mistakenly self-medicated with a product that contained a substance to which it was even more sensitive. The case is of a woman, 36 years old, evolving for 2 months with pruritic eczema, in areas of capillary and nape implantation. She reported progressive worsening. When contact dermatitis was suspected, a contact test was performed using the Latin American baseline series. At 96 hours (D4) there was mild positivity (+) for cocamidopropyl betaine and strong (++) for neomycin. The positive result for neomycin was intriguing, since the observation of the label of products for personal use did not contain it. In view of the test result, after being questioned again, she confirmed the omission of daily self-medication with neomycin ointment. In conclusion, we showed the allergy to hair products. We reinforced the need for an updated baseline series patch test. Finally, we warned about the risk of self-medication. (AU)

A simplified LC-MS/MS method for the quantification of the cardiovascular disease biomarker trimethylamine-N-oxide and its precursors / 药物分析学报

Trimethylamine-N-oxide (TMAO) has emerged as a potential biomarker for atherosclerosis and the development of cardiovascular diseases (CVDs).Although several clinical studies have shown striking associations of TMAO levels with atherosclerosis and CVDs,TMAO determinations are not clinical routine yet.The current methodology relies on isotope-labeled internal standards,which adds to pre-analytical complexity and costs for the quantification of TMAO and its precursors carnitine,betaine or choline.Here,we report a liquid chromatography-tandem mass spectrometry based method that is fast(throughput up to 240 samples/day),consumes low sample volumes (e.g.,from a finger prick),and does not require isotope-labeled standards.We circumvented the analytical problem posed by the presence of endogenous TMAO and its precursors in human plasma by using an artificial plasma matrix for cali-bration.We cross-validated the results obtained using an artificial matrix with those using mouse plasma matrix and demonstrated that TMAO,carnitine,betaine and choline were accurately quantified in 'real-life'human plasma samples from healthy volunteers,obtained either from a finger prick or from venous puncture.Additionally,we assessed the stability of samples stored at-20 ℃ and room temperature.Whereas all metabolites were stable at-20 ℃,increasing concentrations of choline were determined when stored at room temperature.Our method will facilitate the establishment of TMAO as a routine clinical biomarker in hematology in order to assess the risk for CVDs development,or to monitor disease progression and intervention effects.

Development and application of two novel functional molecular markers of BADH2 in rice

BACKGROUND: Fragrance is one of the most important quality traits in rice, and the phenotype is attributed to the loss-of-function betaine aldehyde dehydrogenase (BADH2) gene. At least 12 allelic variations of BADH2 have been identified, and some of these have been applied to rice fragrance breeding using traditional molecular markers and Sanger sequencing techniques. However, these traditional methods have several limitations, such as being very expensive, imprecise, inefficient, and having security issues. Thus, a new molecular marker technology must be developed to improve rice fragrance breeding. RESULTS: In this study, more than 95% of the cultivated fragrant rice varieties belonged to a 7-bp deletion in exon 2 (badh2-E2) or an 8-bp deletion and 3-bp variation in exon 7 (badh2-E7). Both allelic variations resulted in the loss of function of the badh2 gene. We developed two novel SNP molecular markers, SNP_badh2-E2 and SNP_badh2- E7, related to the alleles. Their genotype and phenotype were highly cosegregated in the natural variation of rice accessions, with 160 of the 164 fragrant rice varieties detected with the two markers. These markers cosegregated with the fragrance phenotype in the F2 population. CONCLUSIONS: Two functional SNP molecular markers of badh2-E2 and badh2-E7 allelic variations were developed. These functional SNP molecular markers can be used for genotype and genetic improvement of rice fragrance through marker-assisted selection and will significantly improve the efficiency of fragrant rice breeding and promote commercial molecular breeding of rice in the future.

Choline-betaine pathway contributes to hyperosmotic stress and subsequent lethal stress resistance in Pseudomonas protegens SN15-2

Pseudomonas protegens SN15-2, a typical non-spore-forming rhizosphere bacterium, has excellent biocontrolcapabilities; thus, it is necessary to explore the stress resistance of SN15-2. The choline–glycine betainepathway is considered as an important mechanism by which bacteria adapt to stressful environments. In thiswork, we demonstrated that the expression of the betA and betB genes, which are involved in the choline–glycine betaine pathway in SN15-2, was highly increased by 12-fold and 26-fold, respectively, by hyperosmotic stress and choline treatment. The accumulation of betaine in SN15-2 (5.54 g/L) was significantly higherthan that in the mutants D betA (3.44 g/L) and D betB (2.68 g/L) under hyperosmotic stress and cholinetreatment. Moreover, choline enhanced the growth of SN15-2 greatly, but it did not enhance the growth of DbetB under hyperosmotic stress. Choline combined with hyperosmotic adaptation significantly increased thelethal stress resistance of SN15-2 while the resistance of D betA and D betB was significantly decreased. Thisresearch illuminated a strategy underlying the adaptation to osmotic stress in P. protegens and provided aneffective method to improve the stress resistance of this species, thus provided a theoretical basis for thepractical application of P. protegens SN15-2.

Effect of different pretreatment methods on far-infrared drying characteristics and polysaccharides of Lycium barbarum / 中草药

Objective: To shorten the drying time of Lycium barbarum, increase its shelf life, improve the quality of dried L. barbarum products, and provide technical support for the development of wolfberry drying industry. Methods: Using L. barbarum as experiment material, after being permeated, ultrasonic treating, blanching, ultrasonic treating + permeation and ultrasonic treating + blanchting, far infrared drying test was carried out. The effects of different pretreatment methods on the far-infrared drying characteristics of L. barbarum were studied. Weibull distribution function was used to fit the drying process. The quality and microstructure of dried L. barbarum products were analyzed. Results: After being pretreated, the drying time of L. barbarum was significantly reduced, the drying rate was increased; Compared with L. barbarum without pretreatment before pre-dried, drying time after blanching pretreatment shortened 27.5%, the moisture ratio of the material decreased fastest, and drying rate was the highest. Weibull distribution function can simulate the far infrared drying process of L. barbarum under different pretreatment conditions. The range of r2 and χ2 was 0.991 73-0.999 15 and 8.13 41 × 10-5-8.846 79 × 10-4. The drying characteristics described by scale parameter (α) and shape parameter (β) were consistent with those in the drying characteristic curve. The dried product pretreated with ultrasound + permeation had the smallest color difference (6.756 4) compared with the fresh sample. The polysaccharide content inside the material (991.27 mg/g•DW) was the highest; The microstructure of the dry product obtained by the ultrasound + penetration group had the most voids, and the drying treatment had the least damage to its surface. Conclusion: Ultrasonic and osmotic pretreatment of L. barbarum before drying can not only shorten the drying time, but also improve the quality of its dried products.

Chemical differences between natural and synthetic materials of Bambusae Concretio Silicea based on plant metabolomics / 中草药

Objective: To study the chemical differences between natural and synthetic of Bambusae Concretio Silicea by ultra-high performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Methods: The qualitative analysis of chemical composition was made combined online database and secondary fragmentation cleavage rules, and difference analysis was carried out by the Enhance Peak Find function of PeakView software. Then the converted data was imported into SIMCA-P software to establish an OPLS-DA statistical model and differential secondary metabolites were analyzed. Results: The constituents of Bambusae Concretio Silicea mainly included amino acids, organic acids, alkaloids, glycosides, etc. Among them, 11 components were known, while other 43 compounds were identified for the first time. A total of 14 distinct components, including 2,5-dimethyl-1,3-oxazole-4-carboxylic acid and sucrose, and 12 biomarkers, including 4-(heptyloxy)phenyl-4-(hexyloxy)benzoate, and N-lauryldiethanolamine were identified. Conclusion: This study reveals that the chemical differences between the two herbs are obvious, while betaine and sucrose can be used as the distinguishing indicators. And it provides new ideas and data references for the quality control and clarification of medicinal substances.

Establishment of the Content Determination Method of Betaine in Lycium barbarum and Optimization of the Extraction Method / 中国药房

OBJECTIVE：To establish the content determination method of betaine in Lycium barbarum ，and to optimize the extraction method. METHODS ：HPLC method was used to determine the content of betaine in L. barbarum . The determination was performed on Waters Spherisorb NH 2 column with mobile phase consisted of acetonitrile- 0.01 mol/L monopotassiun phosphate aqueous solution （75∶25，V/V）at the flow rate was 0.7 mL/min. The detection wavelength was 195 nm，and column temperature was 30 ℃. The sample size was 10 μL. Using the content of betaine as index，on the basis of single factor tests ，L（9 34）orthogonal test design were used to select the methanol volume fraction ，extraction time and solid-liquid ratio of betaine in L. barbarum by ultrasonic extraction. The contents of betaine in 10 batches L. barbarum were determined ，and compared with the results of TLC included in 2015 edition of Chinese Pharmacopeia . RESULTS ：The linear range of betaine was 2.035-2 035.04 μg/mL（R2＝0.999 3）. The limits of detection and quantification were 0.410 μ g/mL and 2.051 μ g/mL，respectively. The average recovery were 97.41％-98.86％（RSDs were 0.8％-1.4％，n＝3）. RSDs of precision ，reproducibility and stability （24 h）tests were not higher than 1.2％. The optimal extraction method included solid-liquid ration of 1∶30（g/mL），ultrasonic extraction with methanol for 45 min. The average content of betaine in the extract from the three validation tests was 2.30％（RSD＝0.43％，n＝3）. The contents of betaine in 10 batches L. barbarum were 1.91％ -2.55％ ，which was no significantly different from the results of TLC （1.88％-2.60％）（RE were －1.92％-2.79％）. CONCLUSIONS ：The content determination method of betaine in L. barbarum was established successfully ，and the extract ion process was optinized.

Betaine supplementation improved L-threonine fermentation of Escherichia coli THRD by upregulating zwf (glucose-6-phosphate dehydrogenase) expression

BACKGROUND: The supplementation of betaine, an osmoprotective compatible solute, in the cultivation media has been widely used to protect bacterial cells. To explore the effects of betaine addition on industrial fermentation, Escherichia coli THRD, an L-threonine producer, was used to examine the production of L-threonine with betaine supplementation and the underlying mechanism through which betaine functions was investigated. RESULTS: Betaine supplementation in the medium of E. coli THRD significantly improved L-threonine fermentation parameters. The transcription of zwf and corresponding enzyme activity of glucose-6-phosphate dehydrogenase were significantly promoted by betaine addition, which contributed to an enhanced expression of zwf that provided more nicotinamide adenine dinucleotide phosphate (NADPH) for L-threonine synthesis. In addition, as a result of the betaine addition, the betaine-stimulated expression of enhanced green fluorescent protein (eGFP) under the zwf promoter within a plasmid-based cassette proved to be a transcription-level response of zwf. Finally, the promoter of the phosphoenolpyruvate carboxylase gene ppc in THRD was replaced with that of zwf, while L-threonine fermentation of the new strain was promoted by betaine addition. Conclusions: We reveal a novel mode of betaine that facilitates the microbial production of useful compounds. Betaine supplementation upregulates the expression of zwf and increases the NADPH synthesis, which may be beneficial for the cell growth and thereby promote the production of L-threonine. This finding might be useful for the production of NADPH-dependent amino acids and derivatives in E. coli THRD or other E. coli strains.

Protective effects of betaine-homocysteine methyl transferase on oxidative damaged human lens epithelial cells induced by high homocysteine / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To discuss the protective effects of betaine-homocysteine methyl transferase(BHMT)on oxidative damaged human lens epithelial cells(HLEC)induced by homocysteine.<p>METHODS: HLEC were cultured <i>in vitro</i> and then randomly divided into 3 groups. Normal group:normal cultured HLEC; control group: normal cultured HLEC transfected with empty vector; BHMT gene overexpression group(OE): HLEC transfected with BHMT gene overexpression. All groups were cultured in 10% FCS DMEM +5mmol/L Hcy for 16h. After cultured, BHMT mRNA expression was measured by qRT-PCR and Western blot, the cell proliferation was detected by EdU Assay Kit,The level of ROS and GSH of HLEC were measured by Flow Cytometer and Visible Spectrophotometers. The expression level of of protein(GRP78, Nrf2, Caspase-12)was measured by western blotting. <p>RESULTS: After cultured 16h, cell proliferation ability in OE group was increased by 30.0% compared with NC group(<i>P</i><0.05).The expression of ROS in normal group(89.2043±0.3511)% was obviously higher than OE group(49.5625±0.4502)%, <i>P</i><0.05, GSH activity in OE group was obviously higher than control group and normal group,(<i>P</i><0.05). The expression level of GRP78 in the normal group and the control group was significantly higher than overexpression group. The expression level of Nrf2 in the normal group and the control group was significantly lower than overexpression group. The expression level of Caspase-12 in the overexpression group was significantly lower than that in the control group.<p>CONCLUSION: BHMT <i>in vitro</i> can prevent the oxidative damage of HLEC by high homocysteine, clear the ROS and decrease the ER stress reaction. Apoptosis of lens epithelial cells was inhibited. BHMT plays an important protective role in oxidative damaged HLEC induced by Hcy.

Antifungal Mechanism of Action of Lauryl Betaine Against Skin-Associated Fungus Malassezia restricta

Betaine derivatives are considered major ingredients of shampoos and are commonly used as antistatic and viscosity-increasing agents. Several studies have also suggested that betaine derivatives can be used as antimicrobial agents. However, the antifungal activity and mechanism of action of betaine derivatives have not yet been fully understood. In this study, we investigated the antifungal activity of six betaine derivatives against Malassezia restricta, which is the most frequently isolated fungus from the human skin and is implicated in the development of dandruff. We found that, among the six betaine derivatives, lauryl betaine showed the most potent antifungal activity. The mechanism of action of lauryl betaine was studied mainly using another phylogenetically close model fungal organism, Cryptococcus neoformans, because of a lack of available genetic manipulation and functional genomics tools for M. restricta. Our genome-wide reverse genetic screening method using the C. neoformans gene deletion mutant library showed that the mutants with mutations in genes for cell membrane synthesis and integrity, particularly ergosterol synthesis, are highly sensitive to lauryl betaine. Furthermore, transcriptome changes in both C. neoformans and M. restricta cells grown in the presence of lauryl betaine were analyzed and the results indicated that the compound mainly affected cell membrane synthesis, particularly ergosterol synthesis. Overall, our data demonstrated that lauryl betaine influences ergosterol synthesis in C. neoformans and that the compound exerts a similar mechanism of action on M. restricta.

Halotolerance of Indigenous Fluorescent Pseudomonads in the Presence of Natural Osmoprotectants

Background: Salinity is one of the major factors affecting agriculture. To grow in saline environments, bacteria and plants have to adjust their turgor pressure by accumulating compatible solutes as glycine betaine and proline. Inoculation of plants of economic interest, mainly wheat, by Plant Growth Promoting Rhizobacteria such as Pseudomonas species is an effective biological approach for the recovery of soils affected by salt. Methodology: The halotolerance of indigenous Pseudomonas strains was tested in the presence of high salt concentrations. Under these stress conditions, the effect of natural osmoprotectant molecules elaborated by the halophyte A. halimus was observed. Results: In this study, 3 Fluorescent pseudomonads were isolated from wheat rhizosphere and one from the endophyte of Atriplex halimus. They were identified as P. putida AF2, P. aeruginosa RB5, P. fluorescensRB13 and P. aeruginosa EH4; they exhibited good PGPR activities. The growth of the strains was stimulated in the presence of 100 and 300 mM of NaCl. P. fluorescens CHA0 was inhibited at 500 mM; the remaining strains were affected by 800 mM. Exogenous supply of glycine betaine and proline alleviated the stress. The extract of the halophyte A. halimus restored the growth of 3 strains. NaCl/ 900 mM was strongly inhibitor of all bacteria. The restoration of the growth of P. aeruginosa RB5 and P. aeruginosa EH4 by glycine betaine or proline was significant. No osmoprotectant molecule could overcome stress imposed by 1000 mM. Conclusion: On the basis of their halotolrance and their ability to use natural osmoprotectant to restore their growth, the PGP fluorescent pseudomonads strains tested could be applied as inoculants of wheat for sustainable agriculture in salty soils.

Quality control research of Guanxin Danshen Capsule based on UPLC fingerprint combined with chemical pattern recognition / 中草药

Objective To establish the UPLC fingerprint of Guanxin Danshen Capsule (GDC) and conduct a systemic, comprehensive, and scientific quality evaluation of GDC using a chemical pattern recognition method. Methods Ultra-high performance liquid chromatography instrument and Acquity UPLC® HSS T3 chromatographic column was employed, the separation was performed with the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution, and the detection wavelength was set at 256 nm to establish the UPLC fingerprint of ten batches of GDC. Then, the further quality assessment of the drug was carried out by similarity evaluation, cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA). Results In this research, 75 peaks were recognised as common peaks in the fingerprint, 13 peaks were identified using standard references, they were betaine, succinic acid, tanshinol, protocatechualdehyde, caffeic acid, rutin, galuteolin, rosmarinic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, cryptotanshinone, and tanshinone IIA. The similarity values of the drugs were all above 0.97, indicating a relatively stable quality of the drugs. Little difference was then discovered between the batches of the drug by CA and PCA. Finally, glycinebetaine, rutin, and salvia acid B were recognised as the quality makers using a OPLS-DA method. Conclusion The analysis method established in this study was scientific, accurate, reliable, and simple; The drug quality of GDC could be evaluated systematically and comprehensively using a fingerprint combined with chemical pattern recognition technique. Moreover, it will also lay a solid theoretical basis for the further quality control of traditional Chinese medicine and its preparations at the same time.

Quality assessment of Dandeng Tongnao Soft Capsule based on UPLC- Q-Orbitrap HRMS combined with principal component analysis / 中草药

Objective To establish a quantitative analysis method of multiple active components in Dandeng Tongnao Soft Capsule (DTSC) based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS), and conduct a quality assessment using principal component analysis. Methods The column was Acquity UPLC® BEH C18 (50 mm × 2.1 mm, 1.7 μm) and the mobile phase was consisted of acetonitrile-water (containing 0.1% formic acid) with gradient elution; the information of accurate mass and fragment ions was obtained by the novel “monitored simultaneously for positive and negative ions, full MS scan and automatic trigger secondary mass spectrometry” mode of Q-Orbitrap MS technology to ensure the accurate qualitation and quantitation of the analytes; the results of the contents were then combined with the principal component analysis to achieve the scientific assessment of the different batches of drugs. Results Under the optimized conditions, betaine, danshensu, chlorogenic acid, protocatechuic aldehyde, puerarin, caffeic acid, galuteolin, rosmarinic acid, salvianolic acid A, salvianolic acid B, quercetin, apigenin, senkyunolide A, dihydrotanshinone I, tanshinone I, cryptotanshinone, and tanshinon IIA all showed good liner relationship (r ≥ 0.999 2) in the range of 0.018 1-0.578 4, 5.555 7-177.782 0, 0.018 1-0.580 3, 0.002 8-0.089 2, 0.787 2-25.191 3, 0.000 5-0.016 7, 0.007 2-0.228 9, 0.065 8-2.105 3, 0.304 6-9.747 1, 3.888 1-124.417 6, 0.000 5-0.016 0, 0.000 6-0.018 1, 0.002 5-0.080 1, 0.019 8-0.632 8, 0.032 2-1.031 7, 0.102 8-3.290 0, and 0.044 5- 1.422 9 μg/mL, respectively; The results of the accuracy, repeatability, and stability all reached the standards (RSD ≤ 5%); The recoveries ranged from 98% to 102% and RSDs were below 3%; The analysis results showed that the quality of the most batches was stable, tanshinone I, cryptotanshinone, dihydrotanshinone I, and tanshinon IIA had a great influence on the quality of the drug, which could be monitored to ensure the quality of different batches. Conclusion The methods established in this paper have a high sensitivity and accuracy; the results of the methodology conform to the relevant requirements and the methods can rapidly determinate the multiple active components in DTSC; The research also provides a new scientific basis and reference for the quality assessment at the same time.

Quality assessment study of Compound Xueshuantong Capsules based on UPLC-Q-Orbitrap HRMS combined with principal component analysis / 中草药

Objective To establish a quantitative analysis method of multiple active components in Compound Xueshuantong Capsules (CXC) based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC- Q-Orbitrap HRMS), and make a quality assessment using principal component analysis. Methods The column was Acquity UPLC® BEH C18 (50 mm × 2.1 mm, 1.7 μm) and the mobile phase was consisted of acetonitrile (A)-water (B) (containing 0.1% formic acid) with gradient elution; The information of accurate mass and fragment ions was obtained by the novel “monitored simultaneously for positive and negative ions, Full MS scan and automatic trigger secondary mass spectrometry” mode of Q-Orbitrap MS technology to realize the accurate qualitation and quantitation; Using the quantitative results combined with the principal component analysis to achieve the scientific assessment of the drug in different batches. Results Under the optimized conditions, betaine, succinic acid, salvianic acid A sodium, danshensu, protocatechuate, protocatechuic aldehyde, caffeic acid, rutin, ginsenoside Rg1, rosmarinicacid, salvianolic acid A, salvianolic acid B, wogonin, calycosin, formononetin, astragaloside II, astragaloside I, ginsenoside Rg3, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, and oleanic acid all showed good liner relationship (r ≥ 0.999 0) in the range of 0.009 8-0.314 5, 0.067 8-2.170 7, 0.044 2-1.413 3, 0.059 6-1.907 2, 0.003 3-0.104 4, 0.002 8-0.089 9, 0.001 2-0.038 3, 0.006 3-0.203 2, 0.960 5-30.735 5, 0.022 2-0.709 0, 0.083 7-2.679 5, 0.593 8-19.002 6, 0.000 2-0.005 3, 0.012 3-0.394 4, 0.004 5-0.143 5, 0.009 2-0.293 4, 0.066 0-2.113 3, 0.033 0-1.055 0, 0.004 5-0.145 5, 0.015 9-0.508 1, 0.024 1-0.772 0, 0.009 3-0.297 8, 0.002 5-0.078 8 μg/mL, respectively; The results of the accuracy, repeatability, and stability all reached the standards (RSD ≤ 5%); The recoveries ranged from 98%-101% and RSDs were all below 3%; the analysis results showed that the quality of the most batches was stable, the ginsenoside Rg1, salvianolic acid B, succinic acid, salvianolic acid A, danshensu, and salvianic acid A sodium had a great influence on the quality of the medicine, which could be specially monitored to ensure the quality of different batches of the medicine. Conclusion The methods established in this paper have a high sensitivity and accuracy; The results of the methodology conform to the relevant requirements and the methods can rapidly determinate the multiple active components in CXC. The research also provides a new scientific basis and reference for the quality assessment at the same time.