Simultaneous Determination of the Flavonoids and Limonoids in Citrus junos Seed Shells Using a UPLC–DAD-ESI/MS

Citrus junos seeds (CS) have been traditionally used for the treatment of cancer and neuralgia. They are also used to manufacture edible oil and cosmetic perfume. A large amount of CS shells without oil (CSS) are discarded after the oil in CS is used as foods or herbal remedy. To efficiently utilize CSS as a by-products, it needs to be studied through chemical analysis. Therefore, we developed an ultra-performance liquid chromatography (UPLC)–diode array detection (DAD) method for simultaneous determination and quantitative analysis of five components (two flavonoids and threes limonoids) in CSS. A Waters Acquity UPLC HSS T3 column C18 (2.1 × 100 mm, 1.8 μm) was used for this separation. It was maintained at 40 oC. The mobile phase used for the analysis was distilled water and acetonitrile with gradient elution. To identify the quantity of the five components, a mass spectrometer (MS) with an electrospray ionization (ESI) source was used. The regression equation showed great linearity, with correlation coefficient ≥ 0.9912. Limits of detection (LOD) and limits of quantification (LOQ) of the five compounds were 0.09 – 0.13 and 0.26 – 0.38 μg/mL, respectively. Recoveries of extraction ranged from 97.45% to 101.91%. Relative standard deviation (RSD) values of intra- and inter-day precision were 0.06 – 1.15% and 0.19 – 0.25%, respectively. This UPLC–DAD method can be validated to simultaneously analyze quantities of marker flavonoids and limonoids in CSS.

Development of Molecular Diagnosis Using Multiplex Real-Time PCR and T4 Phage Internal Control to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis from Human Stool Samples

This study aimed to develop a new multiplex real-time PCR detection method for 3 species of waterborne protozoan parasites (Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis) identified as major causes of traveler's diarrhea. Three target genes were specifically and simultaneously detected by the TaqMan probe method for multiple parasitic infection cases, including Cryptosporidium oocyst wall protein for C. parvum, glutamate dehydrogenase for G. lamblia, and internal transcribed spacer 1 for C. cayetanensis. Gene product 21 for bacteriophage T4 was used as an internal control DNA target for monitoring human stool DNA amplification. TaqMan probes were prepared using 4 fluorescent dyes, FAM™, HEX™, Cy5™, and CAL Fluor Red® 610 on C. parvum, G. lamblia, C. cayetanensis, and bacteriophage T4, respectively. We developed a novel primer-probe set for each parasite, a primer-probe cocktail (a mixture of primers and probes for the parasites and the internal control) for multiplex real-time PCR analysis, and a protocol for this detection method. Multiplex real-time PCR with the primer-probe cocktail successfully and specifically detected the target genes of C. parvum, G. lamblia, and C. cayetanensis in the mixed spiked human stool sample. The limit of detection for our assay was 2×10 copies for C. parvum and for C. cayetanensis, while it was 2×10³ copies for G. lamblia. We propose that the multiplex real-time PCR detection method developed here is a useful method for simultaneously diagnosing the most common causative protozoa in traveler's diarrhea.

Multiplex-Touchdown PCR to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the Major Causes of Traveler's Diarrhea

This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the major causes of traveler’s diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum, Glutamate dehydrogenase for G. lamblia, and 18S ribosomal RNA (18S rRNA) for C. cayetanensis. The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in >1×10³ oocysts for C. parvum, >1×10⁴ cysts for G. lamblia, and >1 copy of the 18S rRNA gene for C. cayetanensis. Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum, G. lamblia, and C. cayetanenesis in stool samples.

Sequential magnetic resonance spectroscopic changes in a patient with nonketotic hyperglycinemia / 소아과

Nonketotic hyperglycinemia (NKH) is a rare inborn error of amino acid metabolism. A defect in the glycine cleavage enzyme system results in highly elevated concentrations of glycine in the plasma, urine, cerebrospinal fluid, and brain, resulting in glycine-induced encephalopathy and neuropathy. The prevalence of NKH in Korea is very low, and no reports of surviving patients are available, given the scarcity and poor prognosis of this disease. In the current study, we present a patient with NKH diagnosed on the basis of clinical features, biochemical profiles, and genetic analysis. Magnetic resonance spectroscopy (MRS) allowed the measurement of absolute glycine concentrations in different parts of the brain that showed a significantly increased glycine peak, consolidating the diagnosis of NKH. In additional, serial MRS follow-up showed changes in the glycine/creatinine ratios in different parts of the brain. In conclusion, MRS is an effective, noninvasive diagnostic tool for NKH that can be used to distinguish this disease from other glycine metabolism disorders. It may also be useful for monitoring NKH treatment.

Effects of Early Parenteral Nutrition for Extremely Low Birth Weight Infants

PURPOSE: The object of this study was to evaluate the efficacy of early total parenteral nutrition with early amino acid in extremely low birth weight infant (ELBWI). METHODS: We retrospectively analyzed the medical records of all ELBWIs who were born and admitted to Samsung Medical Center from January 2003 to December 2003 and January 2009 to December 2009 and alive at the time of discharge. Data for nutritional status and morbidities were compared between period 1 (2003, n=22), in which parenteral nutritional support was started gradually over several days and period 2 (2009, n=38), in which parenteral nutrition with amino acid was started as soon as possible after birth. RESULTS: Compared to period 1, birth weight and Apgar score were lower in period 2. The intake amount of glucose, amino acid and total calorie was higher and the level of blood urea nitrogen was increased more from 7th day to 14th day after birth in period 2 when compared than period 1. The weight gain velocity was faster at 7th and 14th postnatal day in period 2. There were no differences in the incidence of necrotizing enterocolitis, moderate to severe bronchopulmonary dysplasia, and severe intraventricular hemorrhage (> or = Gr III) between two periods but, the incidence of periventricular leukomalacia(PVL) was significantly lower in period 2. CONCLUSION: Early initiation of total parenteral nutrition with early amino acid in ELBWIs was beneficial at weight gain with lowering catabolism and increasing anabolism. And it could be related with reducing the incidence of PVL.

Prognosis of Extremely Low Birth Weight infants (1,000 g) Associated with Antenatal Magnesium Sulfate (MgSO4) Exposure / 대한주산의학회잡지

OBJECTIVE: To determine whether magnesium sulfate exposure is associated with a reduced risk of perinatal complications in extremely low birth weight infants (ELBWIs). METHODS: Total 351 inborn extremely low birth weight infants admitted between January 2000 and December 2008 to Samsung Medical Center were divided into the prenatal MgSO4-exposed group (n=76), and control group (n=275), and were retrospectively analyzed. Multiple demographic, clinical characteristics and neonatal complications were compared. RESULTS: Prenatally MgSO4-exposed ELBWIs had significantly higher gestational age, birth weight, and incidence of small for gestational age. In neonatal complications, retinopathy of premature (ROP) treated with laser was less common in MgSO4-exposed group. There were no differences between two groups in the incidence of respiratory distress syndrome (RDS), bronchopulmonary dysplasia (BPD), patent ductus arteriosus (PDA), PDA ligation, intraventricular hemorrhage (IVH), periventricular leukomalacia (PVL), necrotizing enterocolitis (NEC), and mortality. CONCLUSION: Perinatal exposure to magnesium sulfate was associated with decreased incidence of ROP treated with laser.

International travel of Korean children and Dengue fever: A single institutional analysis / 소아과

PURPOSE: Dengue fever occurs in many popular tourist destinations and is increasingly imported by returning travelers in Korea. Since Korea is not an endemic country for dengue fever, pediatricians do not usually suspect dengue fever in febrile children even with typical presentation and exposure history. This study was performed to describe the international travel experiences and dengue fever in Korean children. METHODS: Travel histories were collected based on questionnaires completed by all patients' guardians who visited the pediatric infectious diseases clinic at Samsung Medical Center from January 2008 to December 2008. For patients who were suspected of dengue fever, a serological test was performed. RESULTS: Five hundred and seventeen children visited the pediatric infectious diseases clinic for the first time during this period. About 30% of patients who responded to the questionnaire (101/339) had experienced international travel within the last 2 years. Four patients were diagnosed with dengue fever by serological test. CONCLUSION: Increasing numbers of Korean children visit dengue endemic areas and they may return home with dengue fever. Dengue fever should be suspected in patients who have a travel history to endemic areas.

Optimization of Muffins Containing Dried Leek Powder Using Response Surface Methodology

The purpose of this study was to develop a muffin containing dried leek powder. The complete analysis was conducted using the Design Expert 7 program (Stat - Easy Co., Minneapolis, MN, USA). The leek muffins were produced with varying amounts of leek powder (A), sugar (B), and butter (C). According to response surface methodology (RSM), there were 16 experimental points, including two replicates. The leek muffin formulation was optimized using rheology and sensory analyses. For the results, a quadratic model was applied in determining lightness, volume, hardness, and sensory characteristics, including flavor, texture, and overall quality; redness and height were represented by a linear model. Lightness decreased and redness increased with increasing amounts of leek powder (p<0.001). In addition, hardness displayed significant differences (p<0.001) with increasing amounts of leek powder. The sensory evaluation results showed significant differences for color, flavor, texture, and overall quality (p<0.05). Based on the numerical and graphical methods, the optimal formulation was determined as 8.30 g of leek power, 88.37 g of sugar, and 81.70 g of butter.