Erratum to “Application of AFLP fingerprint analysis for studying the biodiversity of Streptococcus thermophilus” [J. Microbiol. Methods 79 (2009) 48–54

A simple, feasible and effective method of ultra performance liquid chromatography (UPLC) coupled with photo diode array (PDA) were established for fingerprint analysis and simultaneous quantification of three major classes of constitutions including iridoid glycosides, crocins and organic acids of fructus Gardeniae. Extraction method was optimized as 75% methanol ultrasonic extraction for 30 min. Acetonitrile-water (containing 0.2% formic acid) gradient elution on Waters Acquity BEH C18 column (50 × 2.1 mm, 1.7 μm) was used to obtain good chromatographic resolution. 24 characteristic peaks were comprised in the fingerprint common pattern. Among them, seven marked components,geniposide, shanzhiside methyl ester, geinpin, geniposidic acid, crocin I, crocin II, and chlorogenic acid, were quantified. Similarity evaluation, principal component analysis andhierachical cluster analysis were applied to demonstrate the distinction. It was concluded that chemical components of Gardenia jasminoides Eills and Gardenia jasminoides var. radicans Makino from different origins were similar. Other than determination of the contentof geniposide and crocin I, comparison of fingerprint atlas could be considered as a suitable quality control method for fructus Gardeniae. Key words: Fructus Gardeniae, chromatographic fingerprint, multi-component quantification, principal component analysis, hierachical cluster analysis.

IN i960 Cummins and Setzler published an analysis of the finger and palm prints they had collected from mainland aborigines during the 1948 Arnhem Land expedition. The subjects were 41 males and 51 females from Yirrkala in north-eastern Arnhem Land and 43 males and 38 females from Groote Eylandt. The authors gave details of finger and palm patterns but did not report finger ridge counts. Macintosh (1952) analysed the pattern frequencies but not the ridge counts of finger prints from 82 males at Old Beswick and Mainoru in Arnhem Land and from 53 males in the files of the Western Australian Police Department in Perth. Rao (1964a ; b) reported the finger pattern frequencies and total finger ridge counts of 44 males and 40 females from the Juiri, Kulari and Walambi people at Kalumburu on the Western Australian coast north-west of Wyngham. Mader, Parsons, Conner and Hatt (1965) collected finger prints from four tribes in Central Australia (Aranda : 86 males, 102 females ; Pintubi : 64 males, 57 females ; Pitjantjatjara : 139 males, 148 females ; Wailbri : 73 males, 109 females). Their report was confined to pattern indices calculated from the total triradius count of each population and lacked analyses of ridge counts and pattern frequencies. They found some significant differences in inter-tribal pattern indices. Rao (1965) examined finger and palm prints of 61 boys and 40 girls of the Wailbri tribe at Yuendumu Settlement, north-west of Alice Springs in the Northern Territory, and reported the pattern frequencies. The present report is an analysis of the finger and palm prints from 19 males and 19 females of the Kaiadilt and 55 males and 65 females of the Lardili living on Mornington Island in 1966, the total number of tribal members at that time being 94 Kaiadilt and 476 Lardili. The material was collected when the author visited Mornington Island in June, 1966 in association with a psychiatric survey unit from the University of New South Wales. The expedition was supported by the Australiern Institute of Aboriginal Studies. Mornington Island is the largest island in the Wellesley group in the south-east part of the Gulf of Carpentaria, off the Queensland coast (Fig. 1). In 1947 it was inhabited by the Lardili tribe, with a few members from other tribal groups, particularly Janggal from the small Forsyth Island at its southern tip. Bentinck Island some miles to the south-east was the home of the Kaiadilt. For various reasons the Kaiadilt lived a marginal existence and it appears that they had maintained almost complete isolation from the mainland and from the other islands in

Part I: Development of a solid phase extraction coupled with high performance liquid chromatography method for the determination of aripiprazole and dehydroaripiprazole in biological fluid Aripiprazole is the first drug with dopamine partial agonist effect for schizophrenia. Dehydroaripiprazole is its major metabolite. The determination and validation of aripiprazole and dehydroaripiprazole in human serum and urine were performed by a combination of solid phase extraction (SPE) and high performance liquid chromatography (HPLC) in this study. The method includes the following steps: 1) pre-treatment of acid-base solutions for deproteination, 2) application of SPE using an Oasis HLB cartridge for cleaning-up and concentration of the samples, 3) HPLC analysis. The recovery of sample pretreatment step was relatively high with recovery rate of 88.20 - 99.83 %. The optimized HPLC conditions were using a C18 X Terra® column, with an isocratic elution consisted of dipotassium phosphate buffer, pH 8.35, and acetonitrile (40 : 60 v/v) at a flow rate of 1.0 mL/min. The concentration of aripiprazole and dehydroaripiprazole could be determined within 5 minutes. The relative standard deviation (RSD) of the peak area for method repeatability (n = 4) and intermediate precision (inter-day, n = 3) were lower than 0.11 % and 5.16 %, respectively. The calibration curves revealed the method that was linear with concentration range between 50 - 1000 ppb for aripiprazole and 50 - 800 ppb for dehydroaripiprazole. Finally, the validated method was successfully applied to analyze serum and urine samples collected from patients receiving the aripiprazole treatment. The developed method can be used to quantitative determination of aripiprazole and dehydroaripiprazole concentration in patients’ serum and urine for therapeutic monitoring and clinical research. Part II: Fingerprint analysis of rhubarb by capillary electrophoresis and ultra-high pressure liquid chromatography This study used capillary electrophoresis (CE) and ultra performance liquid chromatographic (UPLC) method for chromatographic fingerprint analysis of rhubarb. With the application of chemometric approach, chromatographic fingerprint could be used for species differentiation. Ten common constituents in rhubarb, including aloe-emodine, (+)catechin, chrysophanol, emodine, (-)epicatechin gallate, gallic acid, physcion, rhein, sennoside A and sennoside B, were selected for analytical method development. The optimum micellar electrokinetic chromatography (MEKC) conditions were as followed: 30 mM sodium tetraborate / sodium dihydrogen phosphate monohydrate, 30 mM sodium deoxycholate (SDC), pH 8.6 with 26 % acetonitrile (v/v) as background electrolyte. The optimum condition of UPLC method used a Waters Acquity UPLC BEH C18 column for the separation. The mobile phase was composed of 0.05 % phosphate solution (solution A) and acetonitrile (solution B). The gradient profile was ( solution A: solution B): 0 min, 90 : 10; 25 min, 79 : 21; 35 min, 67 : 33; 40 min, 35 : 65; 45, min 35 : 65. The detector wavelength was set at 254 nm for both methods, and the total analytical time was 21 min for CE and 45 min for UPLC. Sixteen samples of Rheum officinale and Rheum tanguticum collected from various sources were analyzed by optimum analytical conditions. Chromatographic fingerprints of CE were subjected to peak alignment and baseline correction for further similarity test. On the other side, analytical results of UPLC show high precision with flat baseline. Chromatographic fingerprints of UPLC were directly used for Principal component analysis (PCA) and similarity test. PCA shows the chromatographic fingerprints of the two species could be successfully classified. The sample showing the least correlation with the representative chromatographic fingerprint was studied for its DNA sequences. DNA analysis demonstrated the sample to be a hybrid rhizome. The developed CE and UPLC chromatographic fingerprint methods could be applied for the quality control of rhubarb.

The use of chemical analysis of fingerprints as an alternative approach for drug testing has become subject of recent publications. However, the significance of the detection of drugs in fingerprints compared to a background population of non-drug users has not yet been explored. In this research, the main area of research was to determine the forensic potential of detecting cocaine and heroin use through the analysis of fingerprints. Fingerprint samples deposited on paper were extracted using an extraction solution (10% dichloromethane in methanol) and analysed using liquid chromatography – mass spectrometry (LC-MS). This research showed that cocaine and benzoylecgonine were detected in 100 and 94% of natural fingerprint samples (n= 65) collected from drug users, and similarly, heroin and 6-acetylmorphine were detected in 98 and 100% of samples (n = 60). Cocaine and benzoylecgonine were also detected in 13 and 5% of natural fingerprints (n = 98) from a background population of non-drug users. In contrast, heroin and 6-acetylmorphine were detected in 0 and 1% of fingerprints from the background population. For cocaine, a threshold level was required to differentiate fingerprints from drug users and environmental exposure in non-drug users (at a ratio analyte (A) to internal standard (IS) 0.015). The analytes of interest could still be detected in fingerprint samples from drug users after a hand cleaning procedure, however this resulted in a lower detection rate compared to natural fingerprints. In contrast, the analytes were not present in fingerprints collected from non-drug users after handwashing (1% false positive rate for cocaine). Furthermore, cocaine, benzoylecgonine, heroin and 6-acetylmorphine can also be detected in fingerprint samples from dermal contact with the parent drug even after the use of hand cleaning procedures. The detection of illicit drugs in fingerprints is therefore not solely indicative of administration of a drug but does indicate that these analytes are not prevalent in a background population of non-drug users. Additionally, the detection of isoniazid and acetylisoniazid in fingerprint samples from tuberculosis medication showed the potential application of fingerprint testing to monitor adherence to drug treatments. The detection window of isoniazid and acetylisoniazid (<2 days) suggests that a fingerprint may confirm when a patient stops complying to their treatment. This demonstrates that a fingerprint test could confirm non-adherence to treatment, which can be used to help improve treatment plans for patients and improve success rates. Furthermore, the presence of cocaine and benzoylecgonine in urine and saliva collected from drug users was determined using portable mass spectrometry to show the potential for on-site sample analysis for drug testing. Previous work has shown that illicit drugs can be detected in fingerprints, however the suitability of fingerprints for drug testing (namely cocaine and heroin use) has not yet been explored. This research provides information on the influence of the fingerprint sampling strategy, presence of contact residue and transfer of drugs between individuals. These aspects are important to consider in relation to the stages of fingerprint testing and highlight its strengths and weaknesses for further applications (e.g. workplace drug testing, rehabilitation centres and hospitals). To improve fingerprint testing, further work is required on the standardisation of a fingerprint collection procedure, including validation procedures.

Fifteen batches of Glycyrrhizeae Radix standard decoction were prepared for determination of the content of the glycyrrhizic acid and liquiritin, then the transfer rate and the extract rate were calculated and a method was established to analyze the fingerprint by HPLC. According to the measurement of 15 batches of samples,the transfer rate of the glycyrrhizic acid and liquiritin were 59.4%-87.4% and 49.8%-78.9% with extract rate of 29.9%-38.9%. Moreover,10 common chromatographic peaks were determined based on their fingerprint by using similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (TCM)(2012A) .The similarity results of 15 batches of samples were analyzed and compared,and the results showed that the similarity was all higher than 0.9. Fifteen batches of samples,prepared by a standard method,have stable quality and the high similarity.The method displayed good precision,stability and repeatability in fingerprint analysis. Therefore,this study can provide a reference for the quality control of Glycyrrhizae Radix dispensing granules.

The suitability of nappa cabbage juice as raw material to produce freeze-dried probiotics by selected lactic acid bacteria, ( Lactobacillus bulgaricus and Streptococcus thermophillus ) has been investigated. The addition of filler in a form of rice and soybean flour mixture as protective material for viable cells was also assessed. The liquid mixture was fermented at 30 o C for 24 h and sampled at 0 h, 14 h, 24 h and after it was freeze dried at -20 o C. Soybean and rice flour mixture of 10% was added after 14 h of incubation. Titrable acidity, pH, and the number of viable cells were measured during fermentation and after the probiotics had been freeze dried. The carbohydrate, sucrose, lactose, and protein content of each freeze-dried probiotic was also determined. The freeze-dried probiotics analysis showed lactic acid bacteria reduced the pH to as low as 3.69 from 4.50 and increased the acidity from 2% to as high as 6.85%. The filler added to the mixture was found capable of partially protecting the viable cells to as high as 2.45 x 10 5 CFU/mL during freeze drying process. Lactobacillus bulgaricus exhibited slightly higher production of lactic acid than Streptococcus thermophilus even though Streptococcus thermophilus had higher viable cells. The nutrient composition of each freeze-dried probiotic showed closely similar content of carbohydrate and lactose whereas Streptococcus thermophilus had higher concentration of protein and sucrose. This study concludes that nappa cabbage juice could be utilized to produce freeze-dried probiotics.

Objective To analyze the serum related proteomic fingerprints when Lost Goodwill Target (LGT) proteomic fingerprints drifting from negative to positive in the advanced lung adenocarcinoma patients. Methods The serum proteomic fingerprints of 31 advanced lung adenocarcinoma patients whose LGT fingerprints drifted from negative to positive were detected by SELDI and CM10 protein chip. More than 10 % cluster and M/Z values from 11,000+H to 12,000+H was regarded as LGT positive, otherwise as negative. Different fingerprints were screened by Biomarker Wizard 3.1 and Biomarker Wizard software and the decision tree model was established. Results There were 16 statistically different protein peaks when LGT fingerprints drifting from negative to positive, including 10 up-regulation proteomic fingerprints (M/Z:11531, 11483, 11686, 11394, 11822, 11323, 11911, 12450, 5811 and 5709) and 6 down regulation proteomic fingerprints( M/Z: 4126, 13927, 13784, 7001, 1959 and 2741). Conclusion By SELDI and CM10 protein chip detection, up-regulating fingerprints of M/Z 11531, 11483, 11686, 11394, 11822 and 11323 were regarded as the subtype of LGT when it drifting from negative to positive, while up-regulation of M/Z 11911,12450, 5811 and 5709 and down-regulating of M/Z 4126, 13927, 13784, 7001 and 1959 were regarded as the related fingerprints when LGT drifting from negative to positive. The above different fingerprints are constituted of the fingerprints library when LGT fingerprints drifting from negative to positive and it will provide a platform for studying the LGT proteins. Key words: Lung neoplasms; Adenocarcinoma; Spectrometry,mass,matrix-assisted laser desorptim-ionization; Proteomics; Peptide maping

The objective of the work was to conduct a biodiversity study on lactic acid bacteria (LAB) prevailing in the samples of house hold dahi and to evaluate the potency of the isolates to function as active starter cultures. Forty four isolates of LAB (IS-1 to IS-44) were identified from home-made dahi. Biochemical test results proved that isolates were of four different species viz ., Streptococcus thermophilus, Lactobacillus acidophilus, Pediococcus acidilactici and Lactococcus lactis . On comparison of culture activity between different groups of LAB isolates, significantly higher (F<0.01) acid production was observed in group II ( L. acidophilus ) and group IV isolates ( L.lactis ), proving that these two groups of isolates were good starters. With respect to MBRT, group II ( L. acidophilus ) and group III ( P. acidilactici) isolates showed a significantly low dye reduction time proving that they were better starters. Tyrosine value of isolates belonging to group I ( S. thermophilus ) and group II ( L. acidophilus ) were significantly higher. Of the 44 isolates tested 34 percent of the isolates showed the reduction time less than 35 min. proving that all these isolates were “excellent” in their starter activity and ninety percent of the screened LAB isolates showed tyrosine value above 0.2 mg/ml. confirming that most of the selected isolates were having good proteolysis ability. In this study L. acidophilus showed better acid production, proteolytic ability and fast dye reduction. Since the performance of a starter culture is greatly dependant on their activity and purity, it was established that group II isolates was the excellent among screened samples. This study also revealed that the wild strains of LAB existing in home-made dahi are potential source of commercial starter cultures.

A novel spectral fingerprint to discriminate different dry red wines was built using data visualization method. Twelve red wines with different vintages, cultivars and ageing methods from Changli and Shacheng were sampled. Nine fractions of each wine were collected with a reversed-phase C18 column, and then they were lyophilized. The residue of each fraction was resolved with synthetic wine of the same volume with the fraction sample. The transmittance spectra of wines and their fractions were recorded from 190 to 1100 nm. And the spectral data were visualized to show their visual differences directly. Mono-phenols in wine and fractions were analyzed by HPLC-DAD at wavelengths in the range where located the obvious differences of the spectral fingerprints. The results showed that the spectral differences of wine samples lied in the range of 190 to 600 nm. There were obvious differences in visual maps among wines with different vintages, mainly around 520 nm. The visualization differences among wines with distinct geographical origins lay in the F8 maps, and the differences from the aging methods almost cover the whole wavelength range visualized. However, wines from different grape cultivars had the similar visual characteristics. HPLC-DAD identified the possible monophenol groups for the spectral differences at 280, 313, 365 and 520 nm. It was concluded that the visualization of spectral data from 190 to 600 nm could be used to build red wine spectral fingerprint to distinguish dry red wines with different vintages, origins, and ageing methods.

The recombinant plasmid pNCO937 (8.1 kbp) containing a Streptomyces sp. cholesterol oxidase gene was introduced into Streptococcus thermophilus by electrotransformation. Transformation frequency was 7.2 x 10(5) colony forming units/micrograms of DNA. The presence of the cholesterol oxidase gene in S. thermophilus was confirmed with Southern blot analysis using a biotinylated probe. Thin-layer chromatographic analysis showed the expression of the Streptomyces cholesterol oxidase gene resulting in the oxidation of cholesterol to 4-cholesten-3-one. S. thermophilus may be a suitable host for the expression of other genes regulating prokaryotic cholesterol metabolism.

Object: The present study was aimed to assess the anti‑arthritic activity of chloroform fraction of Alstonia scholaris leaf extract against Freund’s complete adjuvant (FCA)‑induced arthritis in rats. Materials and Methods: The anti‑inflammatory activity of various fractions of ethanolic extract of Alstonia scholaris at concentration of 100 mg/kg was studied using the carrageenan‑induced inflammatory models. The chloroform fraction shows significant anti‑inflammatory activity. The chloroform fraction was further studied for anti‑arthritic activity and HPTLC fingerprint analysis. For anti‑arthritic activity, the active chloroform fraction was administered at the concentrations of 50 and 100 mg/kg body weight. The effect of chloroform fraction on liver ALP, ACP and LDH levels of lysosomal enzymes of FCA arthritic animals were studied. Indomethacin and prednisolone (10 mg/kg) was used as standard. HPTLC studies were carried out using CAMAG HPTLC system equipped with linomat IV applicator, TLC scanner; Reprostar 3 and WIN CATS‑4 software were used. Results: The chloroform fraction at 100 mg/kg, showed maximum inhibition (34.16%) of inflammation induced by carrageenan. In FCA‑induced arthritis, the chloroform fraction showed a highly significant reduction in paw volume (50 mg/kg-72.71%; 100 mg/kg-74.35%). The levels of lysosomal enzymes were significantly decreased in the chloroform fraction‑treated groups. Conclusion: The possible mechanism of action of the chloroform fraction of Alstonia scholaris leaf extract may be through its stabilising action on lysosomal membranes. Future studies will provide new insights into the anti‑arthritic activity of Alstonia scholaris and isolation of compound from it may eventually lead to development of a new class of anti‑arthritic agent. Key words: Alstonia scholaris, athritis, chloroform fraction, lysosomal membrane stabilisation

Article Info This research aimed to investigate the viability of probiotic bacteria (Lactobacillus acidophilus LA-5 and Bifidobacterium lactis BB-12) and yogurt bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus) in yogurt during the fermentation, immediately after fermentation and during refrigerated storage (21 d, 4˚C). Also the biochemical characteristics of milk as affected by the commercial 4-strain mixed starter culture were investigated. Storage time affected the viability of all bacterial species. The concentration of lactic acid during the fermentation increased in parallel with the titrable acidity, and the concentration of acetic acid was proportional to the viability of Bifidobacterium lactis. The acetaldehyde level was decreased in the yogurt from day 0 up to the end of the storage. Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus were multiplied considerably during the fermentation. Streptococcus thermophilus could maintain its viability to the highest level, but Lactobacillus delbrueckii ssp. bulgaricus lost its viability rapidly during the cold storage compared to Streptococcus thermophilus. The multiplication and viability of probiotic bacteria were also influenced by the associative strains and species of yogurt organisms. Bifidobacteria counts were satisfactory. The loss of viability for bifidobacteria was gradual and steady during the storage, and they showed good stability during the storage as compared to Lactobacillus acidophilus.

Diving into the deep-end: investigating tropical deep-reef fish assemblages

Objective To study the anti-rheumatoid arthritis mechanism of Sanmiao Pill by using liquid chromatography-mass spectrometry. Methods The rats were randomly divided into control group, model group and medicated group based on weight, 8 rats in each group. The model group and medicated group were treated by intradermal injection of 0.1 ml complete Freund's adjuvant on buttock(s). Rats of medicated group were given Sanmiao Pill solution 0.054 g/ml by gavage, 1 ml/100 g based on weight from the 7th day of experiment for 7 days. And then to score the degree of feet swelling of rats. After 48 hours of the last medication, to collecte the blood samples for metabolic fingerprint analysis. Then analized and processed the non-targeted contour mass spectrometry data (Continuum model) with unsupervised principal component method; identified the maximum metabolic differences between rheumatoid arthritis rats and healthy rats by dimension reduction technic. The differential ions were screened and locked by supervised pattern recognition analysis. Finally, with the help of the automated analysis software database like HMDB, identifiedthe structures and analized the correlated metabolic pathway. Results Compared with the model group, the swelling degree of the feet (2.01 ± 0.19 ml vs. 2.27 ± 0.30 ml) in medicated group significantly decreased (P<0.01). The HMDB data shows that the metabolic profiles of rats were mainly distributed in the control group and the model group, which proved that the rheumatoid arthritis rats treated with Sanmiao Pill had a pullback trend, and the overall metabolic level has a healthy state. With this analysis, 20 blood biomarkers related to rheumatoid arthritis were obtained, which involved the metabolic pathways of aminoacyl-tRNA biosynthesis, Glycine, Serine and Threonine metabolism, Alanine, Aspartic acid and Glutamic acid metabolism, Linoleic acid metabolism, Arachidonic acid metabolism. Conclusions Sanmiao Pill has interventional effect on rheumatoid arthritis rats. The mechanism might be related with metabolic enzymes and metabolic pathways such as Alanine, Aspartic acid and Glutamic acid. Key words: San Miao Pill; Arthritis, rheumatoid; Metabolomics; Biomarkers; Metabolic Pathway; Inflammation; Rats

Minimalist strategies applied to analysis of forensic samples using elemental and molecular analytical techniques – A review