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Objective To establish the method for the simultaneous determination of hypoxanthine, inosine, guanosine and adenosine in Dilong formula granules by HPLC and compare the fingerprints of Dilong formula granules from different manufacturers by HPLC chromatogram. Methods The contents of hypoxanthine, inosine, guanosine and adenosine were determined by Thermo AcclaimTM120C18 column (4.6 mm×250 mm 5 μm). The mobile phase was acetonitrile-water. Gradient elution with flow rate of 0.6 ml/min was used. Column temperature was 25 ℃. Detection wavelength was 254 nm. 10 batches of samples were tested. The HPLC chromatogram were compared and analyzed by using the similarity Evaluation system of chromatographic fingerprint of traditional Chinese Medicine (version 2012.130723). Results The linear ranges for the detection of hypoxanthine, inosine, guanosine and adenosine showed good linear relationships within their own ranges (r≥0.999 9). The average recoveries were 99.20%~102.98% with RSD of 0.26 %~0.71%. The contents of 4 components in 10 batches of samples were 0.740 0~4.457 4 mg/g, 2.132 3~7.805 0 mg/g, 0.325 4~1.596 1 mg/g, 0.537 2~2.222 9 mg/g respectively. The similarity between HPLC chromatogram and control fingerprints of Dilong formula granules from different manufacturers was greater than 0.91. Conclusion The method could be used to determine the contents of hypoxanthine, inosine, guanosine and adenosine in Dilong formula granule. HPLC fingerprints could be used to evaluation the quality in Dilong formula granule. The similarity of HPLC fingerprints from different manufacturer production of Dilong formula granule is high, but 4 contents in composition are difference.
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La hipoxantina y la xantina son biomarcadores metabólicos que resultan de la degradación de las proteínas purinas. Los análisis cienciométricos constituyen una herramienta para estudiar las publicaciones científicas en torno a un determinado tema con la finalidad de determinar tendencias en la literatura. Se realizó un análisis cienciométrico de la producción científica reciente sobre la hipoxantina y xantina en el ejercicio, publicada en la base de datos Scopus durante el período 2016 - 2021. Para la búsqueda en Scopus se utilizaron las palabras clave en idioma inglés: exercise, hypoxanthine y xanthine. Se realizó un análisis cuantitativo, tomando en cuenta los artículos encontrados, así como la información proporcionada por el software VOSviewer. Se identificaron 64 artículos, de estos, 56 fueron de investigación aplicada y ocho de revisión. La categoría de efecto del ejercicio tuvo una mayor cantidad de estudios con 23; dentro de esta se encuentra la subcategoría de metabolismo que presentó 21 artículos. Tanto Estados Unidos como Polonia son los países con mayor número de publicaciones. Existen distintos enfoques y protocolos de ejercicio utilizados para cuantificar la respuesta de la hipoxantina y xantina, así como los perfiles de los sujetos de estudio utilizados como muestra para las investigaciones. La cuantificación de hipoxantina y xantina en el cuerpo es importante para la investigación en el campo de las ciencias del ejercicio(AU)
Hypoxanthine and xanthine are metabolic biomarkers that result from the degradation of purine proteins. Scientometric analyzes constitute a tool to study scientific publications around a certain topic in order to determine trends in the literature. A scientometric analysis was carried out of the recent scientific production on hypoxanthine and xanthine in exercise, published in Scopus database during the period 2016-2021. For the search in Scopus, we used the English keywords exercise, hypoxanthine and xanthine. A quantitative analysis was carried out, taking into account the articles found, as well as the information provided by VOSviewer software. Sixty-four articles were identified, 56 of them were applied research and eight were review. The exercise effect category had a larger number of studies (23). Here there is a subcategory of metabolism that had 21 articles. The United States and Poland are both the countries with the highest number of publications. There are different approaches and exercise protocols used to quantify the response of hypoxanthine and xanthine, as well as the profiles of the study subjects used as a sample for the investigations. The quantification of hypoxanthine and xanthine in the body is important for research in the field of exercise science(AU)
Subject(s)
Humans , Male , Female , Xanthines , Exercise , Muscle Fatigue , Scientific Publication Indicators , HypoxanthinesABSTRACT
The aim of this study was to establish an efficient and stable mouse model of hyperuricemic nephropathy (HN) by testing different modes of administration of potassium oxonate (PO) combined with hypoxanthine (Hx). Animal welfare and experimental procedures were in accordance with the regulations of the Animal Ethics Committee of Guangdong Pharmaceutical University. Male C57BL/6 mice were randomly divided into a control group, a PO+Hx group (i.g.; 100 mg·kg-1·d-1 and 500 mg·kg-1·d-1, respectively), and a PO+Hx group (i.p.; 100 mg·kg-1·d-1, and 500 mg·kg-1·d-1). This HN model was induced by combination of PO and Hx administration once daily for 21 days. The results of serum biochemistry showed that the levels of serum creatinine and 24 h albuminuria were increased compared with the normal group in intragastric administration of PO combined with Hx (P < 0.05), but there was no significant difference in serum uric acid and hepatic levels of xanthine oxidase. The maximum value of serum uric acid and creatinine was 349.3 μmol·L-1 and 26.4 μmol·L-1, respectively, in mice injected with PO combined with Hx. The levels of liver xanthine oxidase and 24 h albuminuria were significantly increased in mice injected with PO combined with Hx (P < 0.01). Pathological data showed that renal tubules were dilated, the epithelial cells of renal tubules were disordered, and the production of collagen fibers, reactive oxygen species (ROS) and lipid peroxidase 4-hydroxynonenal (4-HNE) were slightly increased after intragastric administration of PO combined with Hx mice. Obvious infiltration of inflammatory cells and large area of collagen deposition, with a large amount of ROS and the lipid peroxide 4-HNE were produced in mice injected with PO combined with Hx. Western blot analysis showed that the expression of fibronectin (FN) and urate transporter 1 (URAT1) was increased after intragastric administration of PO combined with Hx in mice and further increased in mice injected with PO combined with Hx. This study demonstrates that injection with 100 mg·kg-1 potassium oxonate combined with 500 mg·kg-1 hypoxanthine establishes a stable and efficient mouse HN model.
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Abstract Hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency is a disorder of purine metabolism responsible for Lesch-Nyhan Disease (LND) and its variants, HPRT-related hyperuricemia with neurologic dysfunction (HND) and HPRT-related hyperuricemia (HRH). The objective of this study was to characterize a cohort of Argentine patients with HPRT deficiency diagnosed in a single center. Results: Twenty nine patients were studied, including 12 LND, 15 HND and 2 HRH. The average onset age was 0.64 years for LND with motor delay as the main manifestation, 8.84 years for HND and 2.5 years for HRH; nephrological manifestations predominated as presenting features in these variants. The average diagnosis age was 3.58 years for LND, 17.21 years for HND and 2.5 years for HRH. Clinical heterogeneity was more evident in HND, even in members of the same family. All patients presented hyperuricemia and no detectable HPRT activity in erythrocyte lysate. The molecular study allowed to identify 9 different mutations in HPRT1 gene from 24 patients (11 independent pedigrees) and to establish genotype-phenotype correlation. In conclusion, this study describes the genotypic/phenotypic spectrum of HPRT deficiency in Argentine patients and highlights the need to increase awareness about the suspicion of these diseases, especially the LND variants with high clinical heterogeneity.
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Objective:To investigate the effect of mangiferin on the mRNA expression of phosphoribosylpyrohoosphate synthetase (PRPS), phosphate ribose pyrophosphate amide transferase (PRPPAT) in liver and hypoxanthine-guanine phosphate transfer enzyme (HGPRT) in brain of hyperuricemic mice induced by potassium oxonate. Method:Hyperuricemic mice were induced through intraperitoneal injection with uricase inhibitor potassium oxonate. The serum uric acid level was determined by the phosphotungstic acid method. The mRNA expression levels of PRPS and PRPPAT in liver as well as HGPRT in brain of hyperuricemic mice were measured by reverse transcriptase polymerase chain reaction (RT-PCR). Result:An intraperitoneal injection with potassium oxonate caused a marked increase in serum uric acid level, compared with normal control group (P-1 was able to significantly reduce serum uric acid levels, compared with hyperuricemic control group (PPConclusion:The hyporuricemic effect of mangiferin might not be related with PRPS, PRPPAT and HGPRT in therapeutic dose.
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OBJECTIVE： To provide reference for improving the quality standard of Pheretima. METHODS： The contents of hypoxanthine and inosine in medicinal material samples were determined by HPLC. HPLC fingerprint of Pheretima was established according to “Similarity evaluation system for TCM chromatogramtic fingerprint” （2012 edition） software， and similarity evaluation was conducted. The determination was performed on Purospher STAR RP-18 endcapped with mobile phase consisted of methanol-water （gradient elution） at the flow rate of 1 mL/min. The detection wavelength was 248 nm， and the column temperature was set at 30 ℃. The sample size was 20 μL. RESULTS： The results of methodological investigation of content determination showed that the linear range of hypoxanthine and inosine were 1.58-31.6 μg/mL （r＝0.999 9）， 5.52-110.4 μg/mL（r＝0.999 8）， respectively. limits of quantify were 0.316， 0.552 μg/mL， respectively； limits of detection were 0.158， 0.110 μg/mL， respectively； RSDs of precision， stability （24 h） and repeatability tests were all less than 2.0％ （n＝6）. Average recovery rates were 103.0％ （RSD＝1.7％， n＝6） and 101.2％ （RSD＝1.2％， n＝6）， respectively. HPLC fingerprint for 15 batches of samples were established， and 8 common peaks were identified. The similarity of HPLC fingerprint of 14 batches of sample with control fingerprint R was higher than 0.900. CONCLUSIONS： The established method for content determination of hypoxanthine and inosine and HPLC fingerprint of Pheretima are simple， accurate and reproducible， and can be used for quality control of Pheretima.
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OBJECTIVE: To establish a quantitative method for determining the contents of uracil, cytidine, hypoxanthine, xanthine, uridine, inosine, and guanosine in Holotrichia diomphalia Larvae by HPLC. METHODS: The HPLC analysis was performed on Waters HSS T3 column(4.6 mm×250 mm, 5 μm). The mobile phase was composed of acetonitrile(A) and water(B), and gradient elution was carried out. The flow rate was 1.0 mL•min-1. The column temperature was maintained at 30 ℃. The detection wavelength was 260 nm. RESULTS: The correlation coefficients of the seven components ranged from 0.999 1 to 1.000 0. The average recovery rates(n=6) were between 91.2% and 97.4%, and the RSDs were between 1.5% and 2.3%. CONCLUSION: The proposed method is simple, accurate and reliable, thus providing basis for comprehensive quality control of Holotrichia diomphalia Larvae.
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PURPOSE: We sought to develop a matrix assisted laser desorption ionization-time of flight (MALDI-TOF)-based, ovarian cancer (OVC), low-mass-ion discriminant equation (LOME) and to evaluate a possible supportive role for triple-TOF mass analysis in identifying metabolic biomarkers. MATERIALS AND METHODS: A total of 114 serum samples from patients with OVC and benign ovarian tumors were subjected to MALDI-TOF analysis and a total of 137 serum samples from healthy female individuals and patients with OVC, colorectal cancer, hepatobiliary cancer, and pancreatic cancer were subjected to triple-TOF analysis. An OVC LOME was constructed by reference to the peak intensity ratios of discriminatory low-mass ion (LMI) pairs. Triple-TOF analysiswas used to select and identify metabolic biomarkers for OVC screening. RESULTS: Three OVC LOMEs were finally constructed using discriminatory LMI pairs (137.1690 and 84.4119 m/z; 496.5022 and 709.7642 m/z; and 524.5614 and 709.7642 m/z); all afforded accuracies of > 90%. The LMIs at 496.5022 m/z and 524.5614 m/z were those of lysophosphatidylcholine (LPC) 16:0 and LPC 18:0. Triple-TOF analysis selected seven discriminative LMIs; each LMI had a specificity > 90%. Of the seven LMIs, fourwith a 137.0455 m/z ion atretention times of 2.04-3.14 minuteswere upregulated in sera from OVC patients; the ion was identified as that derived from hypoxanthine. CONCLUSION: MALDI-TOF–based OVC LOMEs combined with triple-TOF–based OVC metabolic biomarkers allow reliable OVC screening; the techniques are mutually complementary both quantitatively and qualitatively.
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Female , Humans , Biomarkers , Colorectal Neoplasms , Hypoxanthine , Lysophosphatidylcholines , Mass Screening , Mass Spectrometry , Ovarian Neoplasms , Pancreatic Neoplasms , Sensitivity and SpecificityABSTRACT
AIM To establish the HPLC fingerprints of Kangfuxin Liquid (extract of Periplaneta americana L.) and to determine the contents of six constituents.METHODS The analysis of this drug was performed on a TOSOH TSK-GEL ODS column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.07％ acetic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.RESULTS There were twenty-four common peaks in the fingerprints of ten batches of samples (Ⅰ-Ⅹ) with the similarities of 0.932-0.993 (except for sample Ⅰ).Uracil,hypoxanthine,xanthine,inosine,protocatechuic acid and Cyclo (Gly-Tyr) showed good linear relationships within the ranges of 3.460-173.0,3.960-198.0,3.596-179.8,1.338-66.9,3.672-183.6 and 3.552-177.6 μg/mL,whose average recoveries (RSDS) were99.8％ (2.65％),98.0％ (2.55％),99.7％ (1.59％),100.7％ (2.80％),102.0％ (2.09％) and 99.6％ (1.88％),respectively.CONCLUSION This accurate,stable and simple method can be used for the quality control of Kangfuxin Liquid.
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Lesch-Nyhan syndrome(LNS) is a congenital X-linked recessive inherited disorder caused by mutations in the hypoxanthine guanine phosphoribosyl transferase (HPRT) gene.A deficiency of the HPRT enzyme is responsible for the disease.The main clinical manifestation includes hyperuricemia, juvenile-onset gouty arthritis and neurological developmental disorders.Studies have reported there are more than 400 HPRT gene mutation sites, but the incidence of LNS in the Chinese population is extremely low.Here we report a 16-year-old male patient who suffered neurological dysfunction at an early age and gouty arthritis in his youth.DNA of patient and his family members were extracted from peripheral blood lymphocytes.The coding region and the intron-exon boundaries of HPRT gene were sequenced by standard methods.We found a mutation in exon 3 of the HPRT gene of the patient and his mother (Exon3:c.143G>A), which resulted in an arginine to histidine (p.R48H) substitution in the encoded protein.No activity of the enzyme HPRT was detected in the erythrocytes.The same mutation was reported in several European families, but was found in Chinese family for the first time.Clinicians in China have poor experience in diagnosing LNS case, due to the low incidence in China.Therefore LNS screening for infants or adolescents with hyperuricemia, gouty arthritis and neurological dysfunction should be performed.
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OBJECTIVE:To establish a method for the simultaneous determination of uracil,hypoxanthine,xanthine,uridine and adenosine in Eisenia foetida. METHODS:HPLC was performed on the column of Venusil XBP C18 with mobile phase of 50%methanol-0.01 mol/L potassium dihydrogen phosphate solution(gradient elution)at a flow rate of 1.0 ml/min. The detection wave-length was 254 nm with a column temperature at 30 ℃,and the injection volume was 10 μl. RESULTS:The linear range was 0.05-1.60 μg for uracil(r=0.9991),0.05-1.60 μg for hypoxanthine(r=0.9993),0.002-0.010 μg for xanthine(r=0.9991), 0.0075-0.24 μg for uridine(r=0.9999)and 0.0250-0.80 μg for adenosine(r=0.9990);RSDs of precision,stability and accuracy tests were lower than 3%;recoveries were 98.30%-100.76%(RSD=0.8%,n=9),98.68%-100.96%(RSD=0.8%,n=9), 95.16%-97.67%(RSD=0.9%,n=9),96.15%-99.57%(RSD=1.5%,n=9)and 96.39%-101.93%(RSD=1.8%,n=9),respective-ly. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determina-tion of uracil,hypoxanthine,xanthine,uridine and adenosine in E. foetida.
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PURPOSE: Patients show variable responses to chemoradiotherapy (CRT), which is generally administered before surgery for locally advanced rectal cancer (LARC). The aim of this study was to identify molecular markers predictive of CRT responses by analysis of low-mass ions (LMIs) in serum of LARC patients. MATERIALS AND METHODS: LMIs ( 16.0 muM showed significant association with ypStage 0-1 or TRG 4-3 than ypStage 3-4 (p=0.009) or TRG 1 (p=0.024), respectively. In contrast, a significantly lower concentration of PEP was observed in TRG 4-3 compared with TRG 2-1 (p=0.012). CONCLUSION: Findings of this study demonstrated that serum concentrations of HX and PEP, identified using LMI profiling, may be useful for predicting the CRT response of LARC patients before treatment.
Subject(s)
Humans , Biomarkers , Chemoradiotherapy , Enzyme Assays , Hypoxanthine , Ions , Mass Spectrometry , Metabolome , Phosphoenolpyruvate , Rectal NeoplasmsABSTRACT
@#To investigate the therapeutic effect and influence of quercetin and allopurinol on the function of liver and kidney in hyperuricemic rats, male SD rats were administered with the drugs by oral gavage once a day for seven consecutive days throughout the experiment. On the fifth day, the animal model of hyperuricemia was set up by hypoxanthine intraperitoneal injection. The serum was used for assaying the uric acid(UA), alanine aminotransferase(ALT), aspartate aminotransferase(AST), total bilirubin(TBIL), direct bilirubin(DBIL), β2-microglobulin(β2-MG), serum cystatin C(Cys-C), urea nitrogen(Urea)and serum creatinine(Cr)with colorimetry, continuous monitoring, chemical oxidation and enzyme-linked immunosorbent assay. The results showed that quercetin had no effect on the uric acid levels of serum, and that allopurinol reduced uric acid levels in rats significantly(P< 0. 01). The serum levels of AST and ALT in rats substantially decreased compared with normal control group(P< 0. 01), while no significant differences were found in TBIL, and DBIL. Compared with normal control group, β2-MG and Cys-C levels were significantly increased in the other five groups(P< 0. 01), serum levels of Urea and Cr were significantly higher in the allopurinol group compared to the normal control group(P< 0. 01). Modeling and administration group showed mild histopathological changes in rat kidney. The results clearly demonstrated that quercetin had no effect on the uric acid levels of serum, and allopurinol lowered uric acid significantly. There was no significant effect on the function of liver by modeling and administration, however, there was potential impaiment of renal function after modeling. Quercetin did not exhibit a protective effect on renal injury and administration with allopurinol increased kidney damage.
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Objective: To initially establish HPLC fingerprints for analysis of 17 batches of the introdued jujube (Ziziphus jujuba) varieties in order to lay the foundation for scientific evaluation and production quality control of jujube. Methods: TOSOH TSKgel ODS-100V C18 column (150 mm × 4.6 mm, 3 μm) was used. The mobile phase was as follows: methanol and 0.3% KH2PO4 in gradient elution mode. The detective wavelength was 260 nm, the flow rate was 1.0 mL/min, and the separation was performed at 40 ℃. Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica 2004A published by the state Pharmacopeia Committee of China was adopted for the fingerprint analysis on the 17 batches of introdued jujube varieties in Germplasm Nursery of Tarim University. Results: The HPLC characteristic fingerprint of jujube has been established. A total of 22 common peaks were characterized, the peaks appeared uniform and were well-separated. The uracil, hypoxanthine, adenine, cytidine, uridine, cAMP, and cGMP were determined by HPLC method. Conclusion: HPLC chromatographic fingerprint analysis technology can be used for the identification of jujube and its quality control.
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Objective: To establish a UPLC-MS/MS method for the simultaneous determination of nine kinds of nucleosides in Spirodelae Herba, and analyze and investigate the difference of nucleosides ingredients in samples from 13 habitats. Methods: The determination was performed on the Waters XbridgeTM Amide-C18 (150 mm × 4.6 mm, 3.5 μm), the 0.1% formic acid-water (A) and 0.1% acetonitrile-water (B) in gradient elution as mobile phase. The flow rate was 0.5 mL/min. The column temperature was 30 ℃. MS instrument was equipped with ESI+ ion source. Gaining the extracted ion chromatograms, then the peak area for quantitative, principal component analysis (PCA), and hierarchical clustering analysis (HCA) were used to comprehensive evaluation. Results: All nine kinds of nucleoside showed good linearity (r ≥ 0.998 9). The RSD of the precision, repeatability, and stability tests were less than 3.5%. The average recovery rates were in the range of 94.4%-101.9%, RSD were in the range of 1.73% - 3.58%. There are differences in the composition and content from different habitats. The content of 2'-deoxyuridine and 2'-deoxyinosine in the lower levels, cytosine was the lowest, while the contents of uridine, inosine, and xanthine were in higher levels. The largest content was in Spirodelae Herba from Huangshan, Anhui province. Conclusion: The method for the simultaneous determination of the nine kinds of nucleosides is simple, accurate, and reproducible, that will provide the basis for the formulation of herbs Spirodelae Herba for quality control standards.
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Objective To establish an effective method for the determining hypoxanthine, xanthine, uridine and uracil in earthworm in Shanghai Pheretima and Pheretima aspergillum (E. Perrier), contributing to quality control of the medicinal material. Methods Hypoxanthine, xanthine, uridine and uracil were extracted from the earthworms with 0.9% NaCl by ultrasonic and determined by HPLC. The chromatographic conditions were: SORBAX SB-Aq column (250 mm×4.6 mm, 5 μm, Aglient Co.,Ltd), 5 mmol/L KH2PO4 (pH 2.9) as the mobile phase with a flow rate of 1 mL/min,the detection wavelength was 254nm, the column temperature was set at 30℃, and the injection volume was 10 μL. Hypoxanthine, xanthine, uridine and uracil were determined by HPLC at the same time. Results The linear range was 0.5-100 μg (r=0.999 9) for hypoxanthine, with the average recovery being 99.37%, RSD=1.36% (n=6). The linear range was 0.5-100 μg (r=0.993 1) for xanthine, with the average recovery being 91.57%, RSD=1.40% (n=6). The linear range was 0.5-100 μg (r=0.999 9) for uridine, with the average recovery being 95.31%, RSD=1.64% (n=6). The linear range was 0.5-100 μg (r=0.999 9) for uracil, with the average recovery being 100.21%, RSD=1.98% (n=6). Conclusion The current method is reproducible and has satisfactory recovery, and it can be used to determine hypoxanthine, xanthine, uridine and uracil in earthworm medicinal material.
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Creatinine, uric acid, hypoxanthine and xanthine are important diagnostic biomarkers in human urine for gouty arthritis or renal disease diacrisis. A simple method for simultaneous determination of these biomarkers in urine based on reversed-phase high-performance liquid chromatography (RP-HPLC) with ultraviolet (UV) detector was proposed. After pretreatment by dilution, centrifugation and filtration, the biomarkers in urine samples were separated by ODS-BP column by elution with methanol/50 mM NaH2PO4 buffer solution at pH 5.26 (5:95). Good linearity between peak areas and concentrations of standards was obtained for the biomarkers with correlation coefficients in the range of 0.9957-0.9993. The proposed analytical method has satisfactory repeatability (the recovery of data in a range of creatinine, uric acid, hypoxanthine and xanthine was 93.49-97.90%, 95.38-96.45%, 112.46-115.78%and 90.82-97.13%with standard deviation of o5%, respectively) and the limits of detection (LODs, S/N Z 3) for creatinine, uric acid, hypoxanthine, and xanthine were 0.010, 0.025, 0.050 and 0.025 mg/L, respectively. The established method was proved to be simple, accurate, sensitive and reliable for the quantitation of gouty arthritis' biomarkers in human urine samples. The ratio of creatinine to uric acid was found to be a possible factor for assessment of gouty arthritis.
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A validade comercial de sardinhas das espécies Sardinella brasiliensis e Cetengraulis edentulus mantidas sob refrigeração em gelo (0+2°C) foi determinada por parâmetros analíticos físico-químicos, bacteriológicos e sensorial. Nas duas amostras, os teores de Bases Voláteis Totais (BVT) e Trimetilamina (TMA) atingiram o limite máximo recomendado na legislação (30mg N100g-1 para BVT e 4mg N100g-1 para TMA) após 14 e 8 dias de estocagem, respectivamente. O conteúdo de histamina, putrescina e cadaverina se manteve em níveis inferiores a 2.0µg g-1 nas duas amostras durante o período de estocagem. A produção de hipoxantina variou de 0,65 a 2,62µmol g-1 nas amostras de S. brasiliensis e de 1,40 a 2,09µmol g-1 nas amostras de C. edentulus. A contagem inicial de Enterobacteriaceae foi de 3,81log UFC g-1 e 3,82log UFC g-1 atingindo, ao final de 18 dias de estocagem, 6,57log UFC g-1 e 6,87log UFC g-1, nas amostras de S. brasiliensis e C. edentulus, respectivamente. Para as contagens de bactérias heterotróficas aeróbias mesófilas e psicrotróficas, o limite de 7log UFC g-1 preconizado na legislação internacional foi alcançado após o 12° e 8° dias de estocagem nas amostras de S. brasiliensis e após o 12° e 6° dias de estocagem nas amostras de C. edentulus, respectivamente. O método de índice de qualidade sugeriu, para as amostras de S. brasiliensis, um limite de consumo aceitável inferior a 11 e, para as amostras de C. edentulus, um limite de aceitabilidade inferior a 14. Foi proposta a validade comercial de dez dias para a S. brasiliensis e nove dias para o C. edentulus.
Theshelf life of sardines of Sardinella brasiliensis and Cetengraulis edentulus species kept on ice at +2°Cwas determined by physical-chemical, bacteriological and sensory parameters. In both samples, the levels of Total Volatile Bases (TVB) and Trimethylamine (TMA) reached the limits recommended by law (30mg N 100g-1 for TVB and 4mgN100g-1 for TMA) after 14 and 8 days of storage, respectively. The contents of histamine, putrescine and cadaverine remained at levels below 2.0µg g-1 in both samples during the storage period. The hypoxanthine production ranged from 0.65 to 2.62µmol g-1 in samples of S. brasiliensis and 1.40 to 2.09µmol g-1 in samples of C. edentulus. The initial count of Enterobacteriaceae was 3.81logCFU g-1 and 3.82logCFU g-1 reaching, after 18days of storage, 6.57logCFU g-1 and 6.87logCFU g-1, in samples of S. brasiliensis and C. edentulus, respectively. For heterotrophic bacteria aerobic mesophilic and psychrotrophic count the limit of 7logCFU g-1 recommended by international legislation was reached after12 and 8days of storage in samples of S. brasiliensis and after 12 and 6days of storage in samples of C. edentulus, respectively. The quality index method suggested for samples of S. brasiliensis, a limit of acceptable consumption less than 11 and for samples of C. edentulus a limit of acceptability below 14. As a result of this study, we recommend a shelf life of ten days for the S. brasiliensis and nine days for the C. edentulus.
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Lesch-Nyhan disease is a very rare X-linked recessive disorder characterized by mental retardation, spasticity resembling cerebral palsy, choreoathetosis, self-mutilation and hyperuricemia. Self-mutilative behavior is a hallmark of the disease. The underlying defect is a deficiency of hypoxanthine-guanine phosphoribosyltransferase (HPRT). We report on a fourteen-year-old boy, who manifested gouty arthritis and mild renal insufficiency with Lesch-Nyhan disease, lacking self-mutilative behavior in spite of undetectable HPRT activity. Though there were several reports about some cases of Lesch-Nyhan disease in the past Korean literature, the cases were classic forms with definite neurological manifestation. As far as we know, this is the first case of Lesch-Nyhan disease without self-mutilation in Korea.
Subject(s)
Arthritis, Gouty , Cerebral Palsy , Gout , Hyperuricemia , Hypoxanthine Phosphoribosyltransferase , Intellectual Disability , Korea , Lesch-Nyhan Syndrome , Muscle Spasticity , Neurologic Manifestations , Renal InsufficiencyABSTRACT
A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3′ and 5′ ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library ?gt11 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase of Schistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E.coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partial protection vaccine candidate.